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Proceedings of the Research Symposium of Uva Wellassa University, December 15-16, 2011
Development of an Egg Based Sausage

M.K.D.K. Attanayake, D.K.D.D. Jayasena Uva Wellassa University, Badulla, Sri Lanka and A.N. Lalantha Keells Food Products PLC, Ja-Ela , Sri Lanka Introduction The present trend of demand is for ready to serve or ready to cook food items. Sausages have been developed as a convenient product but mostly composed of meat. Present meat and meat based food products may contain various chemicals and preservatives, which is a cause for cancer (Pearson and Gillett, 1996). Therefore, many people nowadays are reluctant to buy these products. On the other hand, absence of strong preservatives in egg-based sausages cuts down health hazards and gives safe access as foodstuffs. Investigations carried out for development of an egg based sausage by replacing the meat component of sausages with eggs has not being attempted yet. Thus, the present study was aimed at producing an egg based sausage for the people who wish to consume egg based products. Egg-based sausage can reach the optimum acceptability and nutritional requirements if developed properly and will provide a balanced diet for the consumer. Eggs are considered as one of the highly nutritious natural products. Eggs naturally possess functional properties like good emulsifying, binding, coagulating and stabilizing abilities, which are essential characteristics in food manufacturing processes (Stadelman and Cotterill, 1996). Therefore, additives with those properties are not required to be added artificially to the manufacturing process of egg based sausages, thus the production cost can be reduced. This study was designed to develop egg based sausages using whole egg powder, egg white powder and locally available high nutritious vegetables such as carrot (Daucus carota), leeks (Allium porrum L.), and mushrooms (Pleurotus species), spices and additives. Fresh eggs are not suitable in sausage production due to several problems. Fresh eggs are difficult to transport since they are bulky, fragile and highly perishable. Moreover, they cause difficulties in stuffing of the sausage mixture into the casing during processing, due to high juiciness and break when fried. Salmonella incidences are also high in raw eggs. Eggs in powder form however, provide a near complete solution to these problems. Dried egg powder could be stored and transported at room temperatures and is quite stable and has a longer shelf life. Egg yolk powder is not suitable to be used in sausages because yolk has higher fat content and low solubility. It also incurs higher import cost and has poor functional properties such as water activity, which is higher in egg yolk powders than in whole egg powders (Joel et al., 2010). Therefore, only whole egg powder and egg white powder could be used in production of egg-based sausages. The main objective of this study was to develop an egg based sausage as an alternative product for traditional meat sausages containing preferable characteristics such as texture, color and taste.
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Methodology The experiment was carried out at the Keells Food Products (PLC), Ja-Ela. The preliminary trials were conducted at first to find out the best combination of egg powder by changing the egg powder ratios. Secondly, the best combination of vegetable oil and water corresponding to the best egg powder combination was chosen. These two experiments were done in order to develop the sausage texture. Then experiments were carried out to determine the best salt level, additive combination and spice combination to adjust the flavor of the sausage. Subsequently, experiments were conducted to find out the chopping level and chamber operations suitable in development of egg-based sausage. Once the satisfactory product was developed, the sensory evaluation, microbiological analysis and chemical analysis were carried out for the samples made using the finalized formula. In order to find out the best recipe which gives better sensory qualities to the sausage, another three samples (T1, T2 and T3) were prepared by changing only the egg powder combination in small quantities in the finalized recipe (i.e. whole egg powder; 15%1.5 and egg white powder; 5%1.5), while keeping the other ingredients constant. Then, in order to select the best percentage combination of whole egg powder and egg white powder, sensory evaluation was carried out with 30 untrained panelists of Keells Food Products (PLC). The sensory evaluation analysis was done using the Friedman nonparametric statistical test. Objective measurements (AOAC, 1995) and proximate analysis (AOAC, 1995) were conducted for all three treatments. Proximate analysis for the data measurement is a combination of crude protein, crude fiber, crude fat, moisture, total solid and ash. Objective measurement was done by analyzing shrinkage percentage, acid value, pH and water holding capacity. For this, samples were taken at regular intervals weekly for two month storage period. Finally data were analyzed using MINITAB and SAS statistical packages. Results and discussion Based on the results of preliminary studies; the best percentage combination was 20% of egg powder, 22% of water, 20% of vegetable oil and 1.4% of salt level. The technical operations of egg-based sausage manufacturing process differed from traditional meatbased sausage. Preferred level of chopping was 6 rounds and the suggested cooking time was 35 minutes appropriate to the core temperature of 76 C of the sausage. No significant difference (P>0.05) was observed among samples for color, appearance, odor, saltiness, and spiciness but texture, taste, juiciness, overall taste and overall acceptability of the samples varied significantly (P<0.05) among samples and also among treatments T1, T2 and T3. According to the Pair wise comparison, texture, juiciness, taste, overall taste and overall acceptability were significantly different (difference between sum of ranks >18.51) between treatments 1-2 or 2-1, and treatments 2-3 or 3-2. And there were no significant difference (difference between sum of ranks among treatments < 18.51) between treatments 1-3, or 3-1. For color, appearances, odour, spiciness and saltiness, there were no significant difference (difference between sum of ranks among treatments < 18.51) between Treatment 1, Treatment 2 and Treatment 3. Based on the results, it can be concluded that the egg-based sausage sample prepared in treatment T3 (whole egg powder with 16.5% and egg white powder 3.5%) had the highest sensory attributes for overall acceptability and overall taste.
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Analysis of variance procedure showed by Duncan group was used for measuring samples which were significantly different (P<0.05) among treatments T1, T2 and T3 with relevant to protein, fat and moisture percentage. Treatment T1 had the highest mean value for protein and lowest mean value for crude fat. Analysis of variance procedure followed by Duncan group showed that treatments from T1, T2 and T3 were not significantly different (P>0.05) in relation to pH, water holding capacity and acid value, but each of these three factors showed a significant increase (P<0.05) with storage duration. The shrinkage percentage was significantly different (P<0.05) among treatments T1, T2 and T3. Salmonella and Escherichia coli were not detected during the storage duration. Staphylococcus aureus counts and TPC did not exceed the specifications in Sri Lankan Standards for the sausage during 60 days. In this period of shelf life, the product was stored at -18C to -30C of temperature. Conclusions According to this research findings, egg based sausages with acceptable characteristics can be developed successfully by; a) Maintaining whole egg powder: egg white powder percentage combination at 20% level. b) Adding 1.4% of salt level into the mixture. c) Using a combination of 22% of water and 20% of fat. d) Chopping only six rounds in bawl chopper. e) Maintaining only cooking operation in the cooking chamber and, f) Maintaining 76 C/35 minutes as the cooking chamber condition. With a lower cost, incorporation of vegetables could be easily done not only to enhance the sensory attributes and nutritional value but also to increase the profit margin of the developed egg-based sausage. Best treatment in this research was T3 (i.e. 16.5% whole egg powder and 3.5% egg white powder) due to its low cost; SL Rs.15.06 per sausage and high sensory attributes. This product was developed without any age discrimination, as it is more health conscious, highly nutritious and more convenient food product. And also it expects a rapid market growth and market stability due to the presence of considerable number of niche markets in Sri Lanka. Without adding any chemical or preservative, shelf life of the egg-based sausage was 60 days at -18 C - (-30 C) with respect to microbiological and physicochemical analysis. References A.O.A.C. 1995. Official method of analysis, 16th ed. Arlington, Association of Official Analytical Chemists, Washington. Joel, N., Udobi., E. Chinweizu, and A. Nuria 2010. Effect of oven drying on the functional and nutritional properties of whole egg and its components. African Journal of Food Science 4(5):254- 257. Pearson, A.M. and T.A. Gillett 1996. Processed Meats, 3rd ed 329- 411. Stadelman, W.J. and O.J. Cotterill 1996. Egg Science and Technology. 4th ed. The Haworth press, Binghamton, New Yolk.
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Development of Ready to Eat Marinated Chicken Parts

M. Laghini, D.K.D.D. Jayasena Uva Wellassa University, Badulla, Sri Lanka A. Kumara and K. Liyanage Institute of Ceylon Agro Industries Ltd., Seeduwa, Colombo, Sri Lanka Introduction The demand for chicken meat and their products in Sri Lanka is growing rapidly, since chicken consumption has no religious barriers. Poultry meat is highly suitable as a raw material for product development due to its light colour and delicate flavour which can be transformed into a wide range of value added foods. Chicken consumption in Sri Lanka is expected to increase to 8 kg per person from the current 5 kg next five years (Cheng Chih Kwong, 2010). Meat items include both ready to eat and ready to cook products. The busy lifestyle of modern day housewives do not allow them adequate time for preparation of food at home. Therefore, this study was conducted with an aim of developing a high quality, safe to eat, marinated chicken parts, which can be stored at frozen conditions and conveniently cooked by the housewives within a short time. A marinade usually tenderizes the meat which is stringy and tough and improves the flavour. Acidic ingredients soften the food, allowing it to absorb the flavours of the sauce and also increase shelf life. These qualities were used in preparing a ready to eat marinade chicken. Materials and methods Marinated Chicken Parts were produced at Ceylon Agro Industries Ltd. in Seeduwa through experiments with sensory evaluation and some laboratory analysis which were conducted at the Uva Wellassa University. Complete Randomized Block Design was used for the experiment. Chicken parts (thighs, wings and drumsticks weighing 2 kg), vinegar, yoghurt, lime, garlic powder (2 g), chilli powder (10 g), salt (8 g), Ins 621- flavour enhancer (10 g), black pepper (2 g), sugar (15 g), mustard cream(1 g), soy sauce (8 g), tomato sauce (2 g), Ins 250preservative (1 g) and water were used to prepare the new product. Preliminary trials and 06 experiments were conducted to select the best levels of ingredients in the recipe of the final product. In the preliminary trials, amounts of vinegar, yoghurt, lime, combination of vinegar and yoghurt, combination of yoghurt and lime and combination of vinegar and lime were used as the variables respectively keeping the other ingredients constant. Then six samples from each recipe were selected and sensory evaluations were conducted to find out the best recipe for marinade. Three experiments were conducted for this purpose and sensory evaluations were carried out using trained panelists of Ceylon Agro Industries Ltd. Then experiments were conducted to select best cooking method using three treatments; cooked at 85 oC for 20 min, cooked at 85 oC for 20 min and baked at 65 oC for 10 min, baked at 65 oC for 35 min and best marinating period was tested using 12 hr, 24 hr and 36 hr periods, respectively through a trained panel. Final sample was compared with a competitive product available in the market through an untrained panel to test the market demand. Thereafter, the samples were tested for moisture, ash, crude protein, crude fat, crude fiber. Microbiological analysis, shelf life determination (pH, WHC) and
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cost determination of the final product were also carried out. All samples were subjected to sensory evaluation and data were analyzed using Friedman analysis in Minitab ver.14. Results of the experiments were analyzed using two-way ANOVA. Results and discussion According to the results of first 3 experiments, vinegar incorporated with yoghurt was selected as the best ingredients for marinating chicken parts. According to the results, marinated solution containing 18 g of vinegar 8 g of yoghurt and other constant ingredients are best for marinating 2 kg of chicken parts obtaining highest scores for texture, taste, aroma, saltiness, pungency, tenderness and overall taste. In the fourth experiment the baked and cooked (baked at 65 oC for 30 min and cooked at 85 oC for 10 min) method was selected as the best cooking method. The cooking method has scored higher for colour, texture, taste and overall acceptability. The fifth experiment showed that 36 hours marinating period was the best which has given better tenderness, pungency and colour. Proximate analysis of the final product contained moisture 64.4%, ash 3.2%, protein 23.5%, fat 3.6% and fiber 5.2%. Crude protein and crude fiber contents of marinated chicken parts were higher than that of the fresh chicken meat since the ingredients in the marinade have added specific nutrient values to the marinated product. Table 1: Storage period with pH and WHC Storage period Control 1st day 1st week 2nd week 3rd week 4th week 5th week 6 5.95 5.9 5.9 5.8 pH Final Sample 5.9 5.9 5.9 5.9 5.9 5.85 Control 0.82 0.85 0.85 0.855 0.855 WHC Final Sample 0.64 0.64 0.64 0.64 0.64 0.64
pH and WHC values in the marinade were better than in the control samples (Table 1). Value of pH was maintained at the same value until 4th week and thereafter it has decreased slightly. WHC value was also maintained at a constant level until 4th week and thereafter showed slightly increased values. Under anaerobic condition, protein degrades to variety of sulphur containing and non-protein nitrogens components which emit gasses and result in decrease in pH and WHC. Table 2: Cost determination for chicken parts Marinated chicken parts Final product Drumstick Rs.291.00 Thigh Rs.168.00 Wings Rs.285.00 Cost per 500 g Control sample Rs. 323.00 Rs. 248.00 Rs. 323.00
Cost determination for chicken parts is given in Table 2. And it reveals that the cost for the new product is less than for what is available in the market.
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Very slight development of total plate count values could be observed with storage time since product has used effective cooking methods. Conclusions Use of vinegar incorporated with yoghurt marinated solution is recommended for making good quality marinated chicken with high acceptability. Keeping quality was better in marinated chicken parts than that of raw product under vacuum packaging conditions. The products are safe for human consumption for more than 6 months under frozen storage -18 oC. References Cheng Chih Kwong, Primus (cited 2011 march 31st), Sri Lankans to eat more chicken Available from <www.lankajournal.com>. Sovann Kin, M., and Wes Schilling 2011. Potassium acetate and Potassium lactate enhance the microbiological and physical properties of marinated catfish fillets. Journal of Food Science. 4:242-245.
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Development of a Low Cost Fish Ball Incorporating Yellow Fin Tuna off Cuts and Deskinned Sword Fish
D.A.B.P. Dasanayaka , D.K.D.D. Jayasena Uva Wellassa University, Badulla, Sri Lanka N. Lalantha Institute of Keells Food Products PLC, Minuwangoda Road, Ekala, Ja Ela, Sri Lanka and S.C. Jayamanne Uva Wellassa University, Badulla, Sri Lanka Introduction The contribution that fisheries resources make to human nutritional needs is significant and may represent the only readily available protein source for people in developing countries. Even small quantity of fish can play a vital role in improving the predominantly cereal based diets of the developing countries. Fish products are comparable to meat and dairy products in nutritional quality. Fish has traditionally been a popular part of the diet in some countries. Today even more people turn to fish as healthy alternative to red meat (Bender, 2005). A recent study has shown that average recovery percentage of expensive cuts of yellow fin tuna (Thunnus albacares) from a medium scale processing factory is approximately 50%. However every recovery percentage can be contrasted depending on the quality of the raw material and nature of the product. The remaining inexpensive off cuts has low market value. Off cuts consists with whole dark muscles, trimmings of the white muscle, tail cuts, and ventral side of the tuna. Tuna trimmings can be purchased at Rs. 200 per kg. The profit margin of food processing companies can be increased while converting these off cuts into value added products. Fish balls can be produced using any fish but the tuna varieties are preferred because meat color and flavor stands up well in finished products. Yellow fin tuna (Thunnus albacares), and Sword fish (Xiphias gladius) are the main species employed. More than two species of fish are usually blended because tuna flesh alone does not give sufficient resilience to the product (Amona, 1965). Materials and methods Experimental work was conducted at the Keells Food Products PLC, Ja-Ela. And the laboratory analysis was carried out at the Uva Wellassa University. Initially, a survey was carried out at the main fish market in Ja-Ela area to identify varieties of fish, usable off cuts, whole sale prices and consumer prices. Subsequent to the preliminary survey selected off-cuts and deskinned sword fish were analyzed for its nutritional quality, dry matter, ash content, crude protein content and crude fat content were analyzed via AOAC methods (AOAC, 2000). Yellow fin tuna (Thunnus albacares) trimmings (histamine content below 25 ppm) and deskinned Sword fish (Xiphias gladius) cuts were obtained from the frozen storage of the Keells Food Products PLC. Frozen fish packages were thawed for 30 minutes to bring the fish meat accessible for cutting. Fish bulk was cut in to 3 cm pieces using a vertical band saw. Then the pieces of fish were minced using a mincer and chopped with a bowl chopper for 2-3 rounds. Chopped fish, pre emulsion, flour, cereal binder, spices and Monosodium Glutamate were then mixed for 4 minutes. Fried onions, green chilies, curry leaves and fresh garlic mixture was
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added to the mixture along with bread crumbs and rusks. The mixture was mixed further for 2 minutes followed by addition of water and fat. Mixture of fish mass was formed into balls using meat ball forming machine. Fish balls were then cooked in a cooking chamber for 40 minutes until the core temperature come to 72 oC (Yu, 1994). Then the fish balls were chilled in order to separate them and vacuum packed in Linear Low Density Polyethylene bags and kept in frozen conditions at -18 oC. The proportions of the ingredients of the fish balls were determined through the preliminary trials by a taste panel comprising seven panelists. Proportions of ingredients were then assessed according to the sensory and textural characteristics. Five combinations of tuna trimmings and sword fish were prepared as given in Table 1 and were tested for color, odour, flavour, spiciness, juiciness, texture and likelihood of breakage during frying and cutting. Based on the preliminary findings, in the final trial, fish balls were formulated with 56% of fish by increasing the amount of tuna trimmings while maintaining the other ingredients constant. Table 1: Percentages of Tuna Trimmings to Sword Fish Treatment Number T1 T2 T3 T4 T5 Tuna Trimmings:Sword Fish 10% : 90% 30% : 70% 50% : 50% 70% : 30% 90% : 10%
A panel of 20 untrained members of Keells Food Products PLC has participated in the sensory evaluation. A 7 point hedonic scale method with a value of 1 corresponded to lowest and a value of 7 to the highest intensity were used to evaluate appearance, colour, odour, spiciness, juiciness, texture, fish taste, after taste and overall acceptability. The samples were coded with three digit random numbers and the order of presentation was made using random permutation. The sensory evaluation was carried out at the Research and Development laboratory of Keells Food Products PLC. All necessary precautions were taken to ensure that each panelist made an independent judgment. Physical analysis was done through folding test and biting test (Lanier, 1992) and cooking loss following Murphy et al. (1975). Objective quality parameters of pH, Water holding capacity (Anjaneyulu, 1989), TBA value (Tarladgis, 1960) and Microbial analysis of Staphylococcus aureus, Escherichia coli and TPC were recorded thrice a week for twelve consecutive weeks. Moisture, crude protein, crude fat, crude fiber and ash content were determined in accordance with Standard AOAC methods (AOAC, 2000). The cost of each item used for the preparation of fish balls was recorded. Statistical Analysis; all measurements were carried out for the experimental design of Complete Randomized Block Design using three replications. The results were reported as the mean standard deviation and subjected to analysis of variance (ANOVA) using SAS V.9.1 statistical software. Differences between the means of fish ball qualities containing different treatments were determined using Duncans multiple range test (DMRT). All statements of significance are based on the probability level of 0.05 (p<0.05).
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Results and discussion The highest estimated median value as well as highest ranks were obtained for the treatment which was having 50% of tuna trimmings and 50% of deskinned sword fish cuts for; Appearance, Colour, Odour, Texture, Fish Taste, After Taste and Overall Acceptability. However, its highest score for juiciness was comparatively low for the seven point hedonic scale. Since the juiciness of above combination is comparatively low, 0.5% of phosphates were added to enhance the juiciness of the selected treatment. There was no significant difference between the treatments for water holding capacity, pH, TBA value. Water Holding Capacity is basically related with the myofibrilla protein of the fish. WHC is reduced in the first six weeks and again increased possibly due to increase of pH which causes more opening to entrap water within myofibrilla proteins. This is a good sign indicates that there is no freeze denaturation with time There was no signicant difference between the first six weeks for WHC in all five treatments but there was a significant difference between first six weeks and ninth week possibly related with increased pH value. There were no significant differences between the initial values of the storage period for pH in the all five treatments. However, a reduction of pH was observed after initial week possibly due to the presence of lactic acid which is resulted from anaerobic metabolism of carbohydrates in the product by the psychrophylic bacteria. It is illustrated that, when the pH increased, the TBA values in all five treatments decreased at the ninth week and the TBA values increased while pH decreased at the twelfth week. It has been reported that haemoglobin (Hb) can show strong pro-oxidant activity for some fish species between pH 6 and pH 7 and it can retard oxidation at pH values above 7 (Richards and Hultin, 2002; Tokur et al., 2004). This may explain why the TBA value decreased when pH increased and vice versa. Reduction in lipid content could be attributed to oxidation of poly-unsaturated fatty acids (PUFA) contained in the fish tissue to produce peroxides, aldehydes ketones and the free fatty acids. However, the rate of fat deterioration was very gradual (Figure 4.5). Fish oil has been found to be more liable to spoilage than other oils due to their greater number of unsaturated fatty acids. The greater the degree of unsaturation, the greater would be the tendency for fat oxidation (rancidity) (Akkus et.al, 2004). There might be risks of rancidity during prolonged storage conditions due to the inherent fatty nature of both tuna and sword fish. The TBA value is widely used as an indicator of the degree of lipid oxidation. The all treatments of developed fish balls were vacuum sealed and kept under frozen condition. Therefore oxidation cannot proceed. But the TBA value in all five treatments increased after sixth week which indicates rancidity of fish balls. The proximate composition of the selected sample which was having 50% of tuna trimmings and 50% of deskinned sword fish was; 61.46% of moisture, 12.98% of crude fat, 15.02% of crude protein, 3.28% of ash and 7.20% of carbohydrates. Cooking yield was 93.22 %. With comparison to the market available fish balls, newly developed fish balls showed higher scores for spiciness, odour and fish taste for its sensory attributes. Scores of folding test was fair and also it gave a fair bite for biting test.
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References Akkus, O., C. Varlk, N. Erkan, and L, Mol 2004. Determination of some quality parameters of fish balls prepared from raw and boiled fish. Turk. J. Vet. Anim. Sci. 44:79-85. Amona, M. 1965. Fish as food. Vol. 3 Academic press, New York. AOAC, 2000. Association of Official Analytical Chemist. Official Methods of Analysis 17th Ed., Washington, DC. Anjaneyulu, Y. L. 1989. Quality of low fat meatballs. Meat Sci. 70:99-108. Bender, 2005. Food Quality and Nutrition. 38:21-30. Lanier, T.C. 1992. Measurements of surimi composition and functional proteins. IBP Press 120-130. Murphy, R.D., G. Serdaroglu, K.J.E Par 1975 . Fish and meat processing technology. Food Engineer. 73:246-252. Richards, L. and H. Hultin, 2002. Chemical and sensory quality changes of fish fingers. Food Chem. 87:523 -529. Tarladgis, A.J., 1960. The effect of heat on protein-rich foods. Food Res. Int., 32:145149.
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Microbiological Quality Assessment of Raw Milk to Identify Sources of Contamination

H.M.G.K.C. Uduwerella, D.C. Mudannayake, A.M.N.L. Abeysinghe Uva Wellassa University, Badulla, Sri Lanka and C.V. Jayasinghe Kotmale Dairy Products (Pvt), Patana, Sri Lanka Introduction: Bogahawatta area is one of the major milk supplying area to Kotmale Dairy Products (Pvt) Ltd, Bogahawatta factory. One of the major challenges faced by Kotmale Products (Pvt) Ltd is the poor microbiological quality of the milk received at the factory. This research was carried out to find out the contribution of contamination sources for milk contamination in Bogahawatta area. Methodology Thirty small holder cattle farmers participated in this study. Farmers were selected using a simple random sampling method. Six samples (two milk samples and four swab samples) were collected from each farmer. Two milk samples (one sample received at the factory under chilling condition & other one received under room temperature) were received. Swab samples were collected from udders of cow, skin of cow, hands of farmer and milking bucket & lid. The time period for receiving milk from farm to the factory and temperature differences occurred during transportation period were recorded. The Total Plate Count (TPC) method was used to enumerate the total aerobic micro organisms present in the samples. Eosin Methylene Blue (EMB) Agar method and Violet Red Bile (VRB) agar method was used for enumeration of E. coli and Coliforms present in the samples, respectively. Karl Pearson Correlation Coefficient method and paired t-test was used to identify the relationship between the contamination sources and contamination of milk. Results The TPC counts revealed that all the factors are not significantly affecting for final milk contamination. According to r values (relationship) of samples, udders of cow is identified as the most related factor (r = 0.322) for the total aerobic micro organisms present in milk. It shows positive low correlation to microbial count in final contaminated milk. Microbial content in milking bucket and lid (r = 0.261) are identified as the second related factor for contamination of final milk while skin of cow, time duration for receiving milk and hands of farmers show negligible relationship to aerobic micro organisms present in final milk. According to the r values (relationship) of E. coli counts it is observed that, skin of cow is the highest related factor (r = 0.593) for E. coli in final contaminated milk. It shows moderate positive relationship to E. coli count in final contaminated milk. Other factors were not significantly affecting final contamination of milk by E. coli. The second factor was identified as the famers hands (r = 0.305). Temperature differences occurred during transportation period showed no relationship to presence of E. coli in final contaminated milk.
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Figure 1: Relationships of contamination sources to Final contaminated milk The Coliform counts showed that all the factors considered had no significant effect on the contamination of final milk. According to the r values (relationship) of Coliform counts, the time duration for receiving milk (r = 0.238) was identified as the factor most related for Coliform development in final milk. It shows positive low correlation to microbial count in final contaminated milk. Temperature difference occurred during transportation period was the second related factor (r = 0.177) while the microbial content in hands of farmer showed no relationship to Coliform presence in final contaminated milk.
Figure 2: Mean values of microbial counts for contamination sources According to the numerical values of total aerobic micro organisms present in samples, skin of cow is identified as the highest contributing source (109650 cfu/mL) for the total aerobic micro organisms present in milk. Microbial content in milking bucket & lid (83497 cfu/mL) are identified as the second significant factor contributing for contamination of final milk. According to the numerical values of E. coli counts it is seen that, skin of cow is the highest contributor (5543 cfu/mL) for E. coli in final contaminated milk. The second factor was identified as the udders of cow (2690 cfu/mL). The Coliform counts numerically showed that milking bucket and lid is the
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highest contributor (3357 cfu/mL) for E. coli in final contaminated milk. The second factor was identified as the hands of farmer (2580 cfu/mL). According to the numerical values of total aerobic micro organisms and E. coli developed during transportation, highest micro organisms development shown as 30 minutes taken for delivering milk to the factory (total aerobic micro organisms - 298600 cfu/ml, E. coli- 29400 cfu/mL). Coliform development shows the highest growth (20400 cfu/ml) in 10 minutes delivery time. According to the numerical values of total aerobic micro organisms developed during transportation, highest total aerobic micro organisms (298600 cfu/mL) development is shown as 3.9 C0 temperature differences occurred at the delivery. For the highest E. coli growth (29400 cfu/mL) it shows at 3.7 C0 temperature differences. Coliform development shows the highest growth (20400 cfu/mL) at 5.6 C0 temperature differences. Discussion According to the results, it shows that presence of E. coli in cow skin significantly affect for E. coli count in final milk. It happened due to farmers practices at milking time. E. coli is indicator for fecal contamination. Before milking they only clean the cow udders not the full body of cow. So the fecal matter can be contaminated to milk. All other factors are not significantly affecting for the micro organisms development in milk. According to the numerical values of each micro organisms present, improper cleaning of udders of cow causes the development of total anaerobic micro organisms present in milk. Micro organisms in cow udder can easily contaminate the milk during the milking time. The numbers of coliforms in milk have increased with the time for receiving milk to the factory. The temperature difference during transportation period also highly affect for coliform development. This can be due to the poor transportation facilities. Conclusions According to the results factors affecting for total plate count in final milk can be orderly listed as microbial content in udders of cow, microbial content in milking bucket & lid and temperature differences incurred during transportation period. For E. coli growth factors can be listed as microbial content in skin of cow, microbial content in hands of famer, microbial content in udders of cow, time duration for receiving milk, microbial content in milking bucket and lid. For Coliform growth, factors are listed as time duration for receiving milk, temperature differences occurred during transportation period, microbial content in udders of cow, microbial content in milking bucket and lid abd microbial content in skin of cow. References Bramley, A. J. and C. H. McKinnon 1990. The microbiology of raw milk. Dairy Microbiology, Ed. R. K. Robinson. Elsevier Applied Science Publishers, London, 1:163-208. Jay, J. M. 1927. Modern Food Microbiology. Aspen poblishers, Inc. Gaithersburg, Maryland, 35-53
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Kurweil, R. and M. Busse 1973. Total count and microflora of freshly drawn milk. Milchwissenschaft, 28:427. Acknowledgment Our respectful acknowledgment goes to all the academic staff members in Department of Animal Science, Faculty of Animal Science and Export Agriculture, Uva Wellassa University, who helped me to achieve research target successfully. Our thanks are also due to all the workers in Kotmale Dairy Products (Pvt), Patana, who gave great supervision for us to conduct my research successfully.
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Development of HACCP Plan for Ice Cream Manufacturing Process at MILCO (Pvt) Ltd.
P.D.A.I. Karunarathne , D.C. Mudannayake Uva Wellassa University, Badulla, Sri Lanka and K. Jayarathne Milco (Pvt) Ltd, Narahenpita, Colombo 05, Sri Lanka Introduction The Hazard Analysis Critical Control Point (HACCP) approach for food safety moves away from testing of final product, and instead emphasizes on raw materials and process control. Control is taken out of the laboratory and in to the processing environment. HACCP provide a structured and systemic approach to the control of identified hazards, which may be biological (microbiological), chemical, physical or combination of the three. A Critical Control Point (CCP) is a raw material, stage, practice or operation within the process where a hazard has been recognized and steps are in place to eliminate, prevent or reduce the possibility of hazard occurring. The application of the HACCP system cover seven principles including identification of potential hazards associated with food production at all stages for processing, manufacture, and distribution until the point of consumption and preventive measures for their control (SLS 1173: 1998). The effectiveness of HACCP depends on the correct application of its principles, combined with other programs (prerequisite programs) such as Good Manufacturing Practices (GMPs), Good Hygiene Practices (GHPs), Standard Operation Practices (SOPs) and Sanitation Standard Operating Procedures (SSOPs). Ice cream, a milk-based product, is a good media for microbial growth due to high nutrient value, almost neutral pH value (pH 6 to 7) and long storage duration (Kanbakan et al., 2004). The quality of ice cream or any food product, can be defined against a wide range of criteria, including for example, the chemical, physical, microbiological and nutritional characteristics. Food or dairy manufacturers aim is to ensure the safety and quality of their products which will satisfy the expectations of the consumers. Considering all above factors, it is critically important to develop a HACCP plan for the ice cream manufacturing process line in MILCO (pvt) Ltd, Narahenpita which practices adequate prerequisite programs to ensure a safe product. Methodology In preliminary studies, all the existing pre requisite programs were assessed and improvements needed were identified. The existing pre-requisite programs were assessed by; preparing an inspection checklist for prerequisite programmes, depth observations of the factory premises and working practices, studying the past and present records, discussing with management staff and observing the available facilities and production equipment. Then ice cream manufacturing process was thoroughly observed and understood by observing the steps of ice cream manufacturing in the factory and then the flow diagram was prepared. Each and every step of the process was
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analysed for potential microbiological, chemical and physical hazards. Identification of critical control points (CCP) was done using a decision tree and critical limits were established for each critical control point. Then corrective actions were established for deviations that observed during monitoring. Finally verification was carried out to develop the HACCP system. Results and discussion Four critical control points were identified for the of ice cream manufacturing process at MILCO (Pvt) Ltd. Those were chilled milk storage in silos, pasteurization, aging and filling. The details of the identified CCPs are given in Table 1. Table 1: Identified Critical Control Points (CCP), Potential hazard, established critical limits and corrective actions were established for deviation that observed during monitoring CCPs Chilled milk storage Hazard Type Biological Potential Hazard Heat resistant enzymes (lipase & protease) and toxins produced by bacteria Critical Limits Storage temperature below 5 C, Maximum storage period of 72 hours Corrective actions Hold the affected silos and, Inform the quality control executive and to decide on disposition Inform quality control executive and re-pasteurize whole batch Maintain the temperature to 5 C by adjusting the circulation of cold water
Pasteurization
Biological
Vegetative pathogens
Ageing
Biological
vegetative pathogens and spores outgrowth
Temperature not less than 90 C and holding time less than 15 seconds Temperature Maintain blow 5 C and minimum holding time not less than 6 hours Microbial count in the filling environment should be zero
Filling
Biological
Microbial Cross contamination
Microbial count (yeast & mould) not be exceed 180, Inform the quality control executive
Conclusions Implementation of HACCP system is a best option for further reducing the risk that is associated with ice cream manufacturing process. Existing pre requisite programs 94
(GMP, SOP, and SSOP) which are already implemented in the factory is not sufficient and improvement should be done accordingly. According to the study, four critical control points were identified. Those are chilled milk storage in silos, pasteurization, aging and filling. Critical limits, monitoring procedures, corrective actions and verification procedures were developed for each identified critical control point, and then the HACCP plan was completed. References Arbuckle, W.S. 1996. Ice Cream. (Connecticut. AVI publishing company, INC). 7142. Forsythe, S.J. and P.R. Hays 1998. Food Hygiene, Microbiology and HACCP. 3 rd Edition (An aspen publication, Gathersburg, Maryland). 236- 300. Sri Lanka Standard, 1173: 1998. Guidelines for the Application of the Hazard Analysis Critical Control Point system (HACCP). Sri Lanka Slandered Institution. Sri Lanka. Acknowledgment We offer our profuse thanks to academic staff members of Animal Science degree program and Faculty of Animal Science and Export Agriculture for giving me great help to conduct this research. We owe our thanks to management staff of Milco (Pvt) Ltd., Narahenpita for providing the necessary facilities to complete this study.
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Development of Chocolate- Malted Whey Beverage Using Liquid Cheese Whey

M.W.I.R. Weerasena, D.C. Mudannayake Uva Wellassa University, Badulla, Sri Lanka and C. Samarasekara Kotmale Dairy Product Private Limited. Mulleriyawa New Town, Sri Lanka Introduction Cheese is one of the most popular milk products which produced by coagulating milk using rennet enzyme and microorganisms. During the cheese manufacturing yellowish liquid that contains all of the nutrition of milk except fat and casein is produced as byproduct or waste material. This liquid is called as whey. Approximately ten pounds of milk are used to produce a pound of cheese and from six to nine pounds of whey is resulted. The whey contains 6-7% solids about half those in milk (Milk and whey powder, 1980). At present, whey is causing a huge problem for the cheese industry. Whey is leading cause for water pollution if whey is discharged to the water source and ultimately this ends up as a major hazard. If whey is unused, its organic nutrients make it a costly pollutant in the countrys sewage and waterways. Biological oxygen demand (BOD) values for cheese whey range from 30,000 to 45,000 milligrams per liter (mg/l) (Milk and whey powder, 1980). Effective and economical methods of utilizing whey are essential if cheese plants are to remain competitive with other segments of the food processing industry. Utilization of whey for beverage production requires few energy resources. The entire whey is utilized no removal of water is necessary. Furthermore, whey can be utilized by small cheese plants for beverage production, since no elaborate or expensive equipment is required. Though the production of whey beverage is cost effective method of utilization of whey, relatively high content of minerals in whey are responsible for undesired salty-sour flavor of whey. This ultimately results with the undesired taste in final product. This can be overcome using chocolate powder and malt extract, since those compounds import not only desirable flavor but also help to increase the nutritive value of final product. The aim of this study was to develop chocolatemalted whey beverage as a solution for the whey disposing problem. In this research, best sugar percentage and suitable percentage of malt extract and cocoa powder were determined separately. Methodology Cheese whey was weighted and heated up to 60 C by using hot water bath. Cocoa powder, sugar, full cream milk powder and malt extract was added to the preheated whey and stirred until all the compounds dissolved completely. The sample was pasteurized using hot water bath for 90 C for five minutes. Sensory evaluation I was carried out to select best sugar percentage (T1-9%, T2-8% and T3-7% sugar) while Sensory evaluation II was carried out to select best combination of cocoa powder and malt extracts (T1- 0.8% Cocoa powder , 0.4%, Malt Extract , T2- 0.4% Cocoa powder ,0.8% Malt Extract, T3- 0.6% Cocoa powder , 0.6%, Malt Extract) Sensory evaluations were carried out using 15 semi trained penalties and data from sensory evaluation was
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analyzed using MINITAB software ver. 14. Friedman statistical method was used to analyze data at the significance level of 0.05. The samples were checked for pH changes and microbiological quality changes at 1 day interval for 10 days during cold storage (4 C). Results Beverage sample that prepared using 8% of sugar was selected as the best sample from sensory evaluation I. In sensory evaluation II, T3 having 0.6% Cocoa powder was selected as the best treatment under 5% significant level. pH changes and microbiological qualities were used as quality parameters for determination of shelf life of final product. Under the microbiological quality total plate count and Coli form count were taken into consideration. As pH of the final product and total plate count of the final product begin to reduce steadily at the 7th day. As a result of that shelf life of product were determined as 7 days under refrigerated conditions. Nutritional value of the final product is given in Table 1. Table 1: Nutritional Value of final product Nutrient Fat Crude Protein Ash Carbohydrate Total Solid Amount g in 100g 1.63 3.45 0.4 14.09 19.57
Conclusions As a pasteurized product final product has higher shelf life than the commercially available milk based pasteurized beverages. Shelf life of the final product is 7 days and can be extended up to 9 days by applying ideal conditions When the nutritional value of final product is considered, it has nearly equal nutritional value to the milk based beverage as shown in Table 1. Sedimentation of cocoa powder was observed with the poor mouth feel in final product and this can be overcome by using food grade stabilizers. Since it is effective and economical solution for whey disposing problem stabilizers were not used for this experiment. Reference Atherton, H.V. and Newlander, J.A. 2000 Chemistry and Testing of Dairy products, Fourth edition, CBS publishers and distributors New Delhi, 395 .
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Identification of Best Pasteurization Temperature Time Combination for Retarding Microorganism Counts in Raw Cream as Ingredient of Butter: Approach to mprove Microbial Quality of Butter
K.A.H.S. Keenavinna, D.C. Mudannayake, A.M.N.L. Abesinghe, Uva Wellassa University,Badulla, Sri Lanka and K. Jayarathne Milco (pvt) Ltd, No.45, Nawala road, Narahenpita. Introduction This paper provides an overview of Best Pasteurization TemperatureTime Combination (BPTTC) for retarding microorganisms in raw cream as an ingredient of butter. BPTTC is an indicator of good quality raw cream as an ingredient of butter. Best pasteurization temperaturetime combination is gaining the idea about good quality raw cream. The quality of raw cream is the most important factor in production of butter. If cream has an increase of microbial count it cant produce butter. In order to determine the qualitymicrobial count is very important. In this study, different pasteurization temperature time combinations were used to retard microorganisms count. This test uses microbial analysis of raw cream by using Total Colony Count (TCC) method and Yeast and Moulds count methods. When cream incorporates high intense heat fat separation occurs. It is not good for the production of quality butter. So when pasteurization temperature time combinations needs to be critically monitored and identified its temperature time combination them unhealthy pasteurization temperature time range can be avoided. An increasing number of people are consuming raw unpasteurized milk. Enhanced nutritional qualities, taste, and health benefits have all been advocated as reasons for increased interest in raw milk consumption. However, science based data to substantiate these claims are limited. People continue to consume raw milk even though numerous epidemiological studies have shown clearly that raw milk can be contaminated by a variety of pathogens, some of which are associated with human illnesses and diseases (Oliver et al., 2009). Food spoilage is an enormous economic problem worldwide. Milk is a highly nutritious food that serves as an excellent growth medium for a wide range of microorganisms. The microbiological quality of milk and dairy products is influenced by the initial flora of raw milk, the processing conditions, and postheat treatment contamination. Undesirable microbes that can cause spoilage of dairy products include Gram negative psychrotrophs, coliforms, lactic acid bacteria, yeasts, and molds. In addition, various bacteria of public health concern such as Salmonella spp., Listeria monocytogenes, Campylobacter jejuni, Yersinia enterocolitica, pathogenic strains of Escherichia coli and enterotoxigenic strains of Staphylococcus aureus may also be found in milk and dairy products (University of West Hungary, 2007). The hygienic production of milk is of the greatest importance for cream, because although most vegetative cells are easily killed by heat treatment, spores are not, and some types, such as B. cereus, can be a cause of spoilage (as well as failure in the
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methylene blue (MB) test). If the spore count of the milk is high (over 100 ml-1), it may be worthwhile reducing this by high speed centrifugal methods. As aerobic spore formers tend to form chains, these are more easily removed than single cell (Robinson, 1990). Most vegetative bacteria cells, yeast and moulds in milk and cream should be killed by pasteurization. However, thermoduric bacteria will survive pasteurization and lactic organisms such as Streptococcus thermophillus, Lactobacillus bulgaricus and Lactococcus lactis are important. Some Micrococcus spp. And some enterococci can also survive pasteurization. Spore-forming thermoduric bacteria survive pasteurization and various aerobic and facultatively aerobic Bacillus spp. can be found in pasteurized milk and cream B. cereus and B. subtilis are proteolytic, whereas B. polymixa is gas forming. B licheniformis and B. coagulans are also found. Clostridium spp. is the main anaerobic apore-forming organism found in pasteurized milk and cream, of which C. butyricum and C. sporogenes are common. Many of the clostridial organisms are proteolytic and/or saccharolytic and those that grow in milk also form gas. Frazier and Westhoff (1988) recorded that heat resistant lactic acid bacteria may spoil pasteurized products through the production of lactic acid, provided the storage conditions are favourable, i.e. the temperature is high enough. When storage temperatures are unfavourable for lactic bacteria, coliforms (the result of post-pasteurization contamination) may produce hydrogen, carbon dioxide, ethanol, formic acid, acetic acid and some lactic acid. In the absence of competition from lactic acid bacteria Bacillus spp. and Closridium spp. will causes spoilage, the former mostly producing lactic acid and the latter often producing butyric acid, hydrogen and carbon dioxide (Early, 1998). Methodology In this project, milk and milk products move through several hands from production to consumer. Hence the assessment of overall efficiency of handling and processing of the products is a criterion in the evaluation of the quality delivered to the consumer. This is achieved by testing the samples of milk, milk products, water, swabs and rinse of dairy utensils in the microbiology laboratory. Each pasteurization temperature time combination were used to five samples (cream) and the micro biological analysis was done. Also one temperature has two time changes. Good quality raw milk was tested by using Resazurin test. Total Colony Counts (TCC) and Yeast and moulds microbiological analysis were done to the find out the best pasteurized cream sample. Results and discussion In this study, when pasteurization temperature was increased and TCC level and Yeast and mould counts level were reduced. Also when time period were increased and also results were obtaining in lowest counts of microorganisms level. Bulk collection of milk is common practice to be stored in creameries at 5 oC for up to 48 hours and even sometimes longer. The change from churn to bulk milk collection has resulted in a change in the microflora of raw milk, with an increase in the level of psychrotrophs. There is normally no problem with milk quality for milk so held, although psychrotrophs can grow very slowly at below 5 oC. These are generally biochemically active against fat and protein, but do not usually ferment lactose to lactic acid. Thus cold stored milk does not sour but can develop taints, and although the organisms are nearly all killed by pasteurization, they produce enzymes which can survive pasteurization and continue to produce changes in the pasteurized product 99
(Robinson, 1990). The raw cream is usually held at about the separation temperature (e.g. 40 oC) during the standardization. Furthermore, the cream may be contaminated by inadequately disinfected process plant or the skim milk may not be of good quality (Robinson, 1990). Also high temperature pasteurization fat separation occurs and that problem mainly caused to the final product quality. So, high intended heat wants to avoid because of the fat separation. Also high intended heat has economic losses. Because of heat generating fuel is diesel and high heat wants to get much fuel burning. It is major threat for production of customer based product. If price is high product also fails in the market and the customer penetration is reduced. When free fatty acids of the cream are increased more than 0.37 % then it cant produce butter. So it converts in to ghee. Low temperature has more free fatty acids level so it pasteurization temperature time combination cant produce butter. If that pasteurized cream is converted to butter, its shelf life is reduced and also off flavor is developed. High temperature pasteurization combinations produce low free fatty acids cream and it is good for production of butter. All milk and products made from it, such as cream, become contaminated by micro organisms from the udder, or from the cow, or during the milking process. The original flora in this sense consists mainly of lactic and other streptococci, micrococci, corynebacteria, and aerobic and anaerobic spore forming bacteria. Conclusions P < 0.05, and all the temperature time combinations were significant with 75 oC 30 minutes pasteurization temperature - time combination. When temperature increases with more than 75 oC temperature (e.g. 80 oC) fat separation occurs and it can affect the final product quality. So, the best pasteurization combination is 75 oC 30 minutes. This best pasteurization combination there have no any fat separation and also economic viable temperature time combinations. References Adams, M.R., M.O. Moss 1996. Food microbiology. 104 112. New Age International (Pvt) Ltd. ANON, 2009. Pasteurization definition and methods. IDFA, June 2009. ANON, 1953. Milk pasteurization. World health organization, Geneva, 1953. Early, R., 1998. The technology of dairy products. 2nd ed., Blackie academic & professional, London. Oliver, S.P., K.J. Boor S.C. Murphy and S.E. Murinda 2009. Food safety hazards associated with consumption of raw milk. Food borne pathogen and disease. 793 - 803. Robinson, R.K. 1990. The Microbiology of Milk Products. 2nd ed., Boca Raton, New York. Spreer, E. 1998. Mlik and Dairy Product Technology. MARCEL DEKKER INC, New York.
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A Study on the Mangrove Crabs in Batticaloa District for Potential Export Market
A. Krithika and S.C. Jayamanne Uva Wellassa University, Badulla, Sri Lanka Introduction Mangrove crabs live in association with mangrove forests and belong to many different species. They have been shown to be ecologically very significant animals which keep much of the energy within the forest by burying and consuming leaf litter. The edible mangrove crab Scylla serrata is a well known commercial commodity considered to be among the tastiest of crab species and have a huge demand in South Asian countries. Most of the edible crabs belong to the family Portunidae and are swimming crabs but there are some grapsid crabs which are true dwellers of the mangrove forests that are edible. Grapsid crab, Episesarma sp. is such a delicacy consumed by the Thai people and by Chinese people in their cuisines (Ng and Sivasothi, 1999). Some Uca species and sesarmid crabs such as Perisesarma species which has deep red pincers and iridescent blue or green band across the face are considered as ornamental animals due to their beautiful colourations (Nyawira and Methiga, 1834). Sesarma bidens is another mangrove crab which is bred in aquariums as an ornamental animal (www.aquaticcommunity.com). Extensive mangrove forests are found in association with lagoons and estuaries of Sri Lanka but the studies on their fauna is limited (Priyadarshani, et al., 2008). The crabs dwelling in these mangrove environments are anticipated to have much potential to be utilized in various industries if they could be bred in captivity. Crabs with ornamental value can be bred in captivity and can take up to the ornamental aquaculture markets. The present study was carried out with an aim of finding the diversity and the export potential of the mangrove crabs in the Batticaloa District, Sri Lanka. Methodology Three mangrove areas in the Batticaloa (7430N, 81420E) district of Sri Lanka were randomly selected as study sites. The selected mangrove forests are situated in the Grama Sewaka divisions of Mankerny, Saththurukkondan and Kokkaddicholai. Three belt transects with a width of 10 m and a length of 50 m were laid perpendicular to the lagoon shore at each site and were subdivided in to 10x10 m2 plots. Alternate plots starting from the lagoon shore were selected for detailed studies totaling three sampling plots of 10 m x 10 m per one belt transect. Each sampling plot had three systematic sampling units of 1x1 m2 size and all the crabs in such plots were collected, identified and counted. The crabs were collected by digging the soil up to the water table until the crabs are caught and were picked by hand. The crabs were handled with caution to avoid autotomy which causes difficulties in identification. Soon after the collection the crabs suitable for taxonomic studies were stored in a freezer. The samples for further laboratory references were stored under 4 oC to prevent changes in the physical parameters attributable to the microbial degradation. The crabs were identified to the species level using the available keys and diagnostic characters described by the former taxonomists (Ng and Sivasothy, 1999; Priyadarshani, et al., 2008).
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The environmental variables were analyzed quantitatively to find out the distribution of crabs in relation to environmental parameters. The physico-chemical parameters; air and soil and water temperature, conductivity, salinity and pH were measured in each plot to find out their habitat preference. The crabs species collected from the study sites were recorded as edible crabs considering their food value, large size and their acceptance by the Food and Agricultural Organization (FAO) as harmless to human consumption and as ornamental crabs considering their eye catching attributes and minor sizes. The experiment was designed as a complete randomized block design and the data were analyzed using two- way ANOVA. Results and discussion Twelve species of brachyuran crabs of which three species belonging to family Ocypodidae, and nine belonging to family Grapsidae were identified from the three mangrove forests in the Batticaloa district. The species Episesarma versicolor, Episesarma mederi and Varuna litterata are accepted as edible species according to FAO and Uca annulipes, Uca chlorophthalmus, Metapograpsus oceanicus, Parasesarma plicatum, Perisesarma eumolpe, Metasesarma obesum are identified as ornamental crabs. The crabs, Cleistostoma merguiense, Metopograpsus thukuhar and parasesarma sp. were identified as not belonging to above two categories. None of the crabs show a significant relationship with the salinity while three Ocypodid crabs, Uca annulipes, Uca chlorophthalmus, Cleistosoma and Metapograpsus thukuhar have shown a significant negative correlation (P<0.05) with the pH. Episesarma versicolor showed a positive correlation with the water and air temperature (P<0.05) while Perisesarma eumolpe showed a negative correlation (P<0.05) with the water and air temperature. Episesarma mederi and Varuna litterata showed a positive correlation with the conductivity while Metasesarma obesum showed a negative correlation. Metapograpsus oceanicus preferred highest mean salinity level (31 ppt) while Episesarma versicolor preferred low salinity (8 ppt). Highest abundance of crabs was observed from Mankerny where average abundance was 59/m2 followed by 24/m2 in Batticaloa and 18/m2 in Kokkaddicholai. Highest species richness was recorded from Mankerny (10 species) followed by Batticaloa (03 species) and Kokkaddicholai (02). Species diversity in Mankerny was very high (H = 2.14) compared to those of Batticaloa (H=0.72) and Kokaddicholai (H=0.52). Conclusions It is evident that crabs of Batticaloa area specially in Mankerni have a good export market potential with nine out of twelve species have either a food value or ornamental value. The findings of this research will pave way to emerge various new industries related to mangrove crab fisheries, culture, breeding and processing. Furthermore, such investigation will help many further scholarly progressions regarding taxonomy and diversity. However, accessing of export market has to be done only after the successful breeding programmes for the crabs have been established, to avoid extinction of these valuable resources. It is also crucial to establish government regulations regarding use of mangrove crabs for such purposes.
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References Ng, Peter K. L. and N. Sivasothi 1999. A Guide to the Mangroves of Singapore II (Animal Diversity). Singapore Science Centre. 168. Nyawira, A. and K.M.F.R.I.Muthiga 1834 Edible crabs of Kenya. Aquatic Bulletin 3: 61-65. Priyadarshini, S.H.R., S.C.Jayamanne and Y.N. Hirimuthugoda 2008) Diversity of mangrove crabs in Kadolkele, Negombo eatuary. J. Aquat. Sci. 13:109 121. Acknowledgement The authors wish to extend their sincere gratitude to Prof. P.K.L. Ng of the National University of Singapore for the support given in confirming all species and identifying some species.
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Development of Ginger Flavoured Pasteurized Milk with Incorporation of Ginger (Zingiber officinale) Extract and Sugar
N.M.P.K. Upananda, A.M.N.L. Abesinghe and D.C. Mudannayake Uva Wellassa University, Badulla, Sri Lanka. Introduction The Sri Lankan dairy industry is important and has tremendous potential in developing the economy in the country. Since centuries, milk is used for direct consumption as well as for making various products. With the advent of new processing techniques, many products especially such as pasteurized milk were added. Within this milk types, flavoured milk remained highly demanded. However, there was no ginger flavoured milk type among the flavoured pasteurized milk, which has antioxidant, antimicrobial and anti-tumour effect with many other medicinal values. Therefore, this research has focused to add value to flavoured milk by incorporating ginger extract. Methodology Ginger (Zingiber officinale) rhizomes were washed with water and soaked for 12 hours. Then, they were sliced (2 3 mm) and grinded. This mixture was heat treated for 5 minutes at 100 oC and filtered to obtain the ginger extract. Then, samples of ginger flavoured milk were prepared by incorporating ginger extract and sugar in to cow milk. There were 9 ginger flavoured milk samples by changing the volume of ginger extract as 8 mL, 10 mL and 12 mL and sugar percentages as 5%, 7.5% and 10%. For the preparation of each flavoured milk sample, relevant ginger extract volume and sugar % were added in to 100 mL of milk separately and they were pasteurized at 72 oC for 15 seconds. Plain pasteurized milk samples were used as the control. An evaluation panel comprising 5 trained panellists and 10 untrained panellists was used for first three sensory evaluations. Sensory attributes; appearance/colour, aroma/smell, pungent/ginger flavour, sweet taste, overall flavour and overall acceptability were tested using 9 point Hedonic scale (from 1-extremely dislike to 9-extreamly like). These sensory evaluations were conducted by changing the ginger extract volume while keeping the sugar % constant. With each evaluation, three best combinations were selected. Selected three samples were produced again using the same procedure and the best combination was selected by a sensory panel comprising 39 untrained members. The selected milk sample was packed in LDPE bags and stored below 4 oC for further analysis. The shelf life was determined by analysing titratable acidity (TA) and microbiological evaluations (Total Plate Count and E. coli). Proximate analysis was carried out by measuring the fat%, protein content, moisture% and ash content. Data obtained from sensory evaluations were analysed by Friedman test. Results and discussion From the first three sensory evaluations, 8 mL ginger extract volume was selected for all three sugar %. From the final sensory evaluation, ginger flavoured milk sample with 8 mL ginger extract volume and 7.5 % sugar was selected as the best sample. Results from microbiological analysis indicated that SPC of ginger flavoured milk sample did not change (p>0.05) throughout the tested period (11 days). The TA of selected ginger flavoured milk sample behaves similarly. Milk fat, protein, moisture, total solids and
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ash content for selected milk were 3.6%, 291.6 mg, 89.9%, 10 % and 0.02 g respectively. According to the results obtained from final sensory evaluation (Figure 1), there were no difference (p>0.05) in appearance/ colour among four samples. This may be due to the addition of ginger extract and sugar in law quantities which may not contribute to significant colour variation.
Appe aranc e/col our 8 Over all acce ptabi lity 6 4 2 0 Over all Flav our Swee t taste Pung ent/G inger flavo ur Aro ma/S mell
40 35 30 25 20 15 10 5 0 0 1 4 6 8 11 Storage period (days) Ginger incoporated milk Control
5% sugar 10% sugar
7.5% sugar Control
Figure 3. Variation in the sensory attributes of milk samples
Panelists prefer milk samples with sugar percentages as 5%, 7.5% with respect to aroma/smell than other milk samples. All three ginger incorporated milk samples were preferred by the panelists than the control. This may be due to the pleasant aroma of ginger which caused by more than 70 constituents present in steam volatile oil. According to the findings of Purseglove et al. (1983) sesquiterpene hydrocarbon zingiberene is predominates in ginger and accounts for 2030% of the volatile oil obtained from dry ginger. The most preferred milk sample with respect to pungent/ginger flavour is 7.5% sugar and 8 mL of ginger extract incorporated milk sample. All three ginger incorporated milk samples were preferred by the panelists than the control. These results are in accordance with the findings of Puengphian and Sirichote (2006). He explains that the main pungent ginger constituents are oleoresins such as 6-, 8- and 10-gingerols and 6shogaol. These constituents are well dissolved in non polar solvents such as milk fat.
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SPC 103 mL -1
Figure 4. variation of SPC in ginger flavored milk and control
Milk fat was evenly distributed all over the milk serum after the homogenization. Therefore, these oleoresins were uniformly distributed in the milk. Results from microbiological analysis indicate that SPC (Figure 2) of the selected ginger flavoured milk sample did not change (P>0.05) throughout the tested period (11 days). The TA of selected ginger flavoured milk sample behaves similarly. However, SPC of the control exceeded the SLS specifications for pasteurized milk (30000 cfu/mL) at 7th day of refrigerated storage. According to the above results, incorporation of ginger extract enhanced the shelf life of pasteurized milk. This may be due to the antibacterial effect of ginger. Malu et al. (2008) found that the antibacterial activity and inhibition activity of ginger extracts could be due to the constituents such as sesquiterpenoids, zingiberene, sesquiphellandrene, bisabolene, farnesene and trace monoterpenoid fraction, (sesquiphellandrene, cineol and citral). Azu et al. (2007) also described that, these components have antibacterial and gastrointestinal tract motility effects. Ginger has the capacity to eliminate harmful bacteria, such as E. coli, responsible for most of the diarrhoea, especially in children. Ginger eases both diarrhoea and constipation; hence it should have impact on the growth of Bacillus cereus, which mainly causes diarrhoea and nausea). Conclusions Ginger flavoured pasteurized milk can be produced by incorporation of ginger extract. Most preferable combination is 8 mL of ginger extract and 7.5% of sugar content for 100 mL of milk. The product shelf life was expanded rather than market pasteurized milk types. The product might be having some medicinal advantages, which is derived from ginger. References Azu, N. C. and R. A. Onyeagba 2007. Antimicrobial Properties Of Extracts Of Allium cepa (Onions) And Zingiber officinale (Ginger) On Escherichia coli, Salmonella typhi And Bacillus subtilis, Internet Journal of Tropical Medicine, 3 (2):15402681. Gao, D. and Y. Zhang 2010. Comparative antibacterial activities of extracts of dried ginger and processed ginger. Pharmacognosy Journal 2 (15): 41-44. Malu, S.P., G.O. Obochi, E.N. Tawo, and B.E. Nyona 2008. Antibacterial activity and medicinal properties of ginger (Zingiber officinale)., Global Journal of pure and applied sciences, 15 (3): 65-368 . Puengphian, C., and A. Sirichote, 2008. [6]-gingerol content and bioactive properties of ginger (Zingiber officinale Roscoe) extracts from supercritical CO2 extraction. Asian Journal of Food and Agro industry. 29-36. Purseglove, J. W., E. G. Brown, C. L. Green, and S. R. J. Robbins 1981. Spices 2: 389421. Sri Lanka Standard., 1983. Specification for raw and proceed milk (SLS 181:1983). Bureau of Ceylon standards. 637.141.
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Application of Green Supply Chain Management Approach for a Community Based Dairy Factory
S.V.G.A. Samaranayake, D.C. Mudannayake, A.M.N.L. Abesinghe, Uva Wellassa University, Badulla, Sri Lanka and W.U.S. Alwis Industrial Development Board, Katubedda, Moratuwa, Sri Lanka Introduction This paper provides an overview of Green Supply Chain Management (GSCM) approaches for a community based dairy factory. GSCM is an emerging field that out of the traditional supply chain perspective. Greening the supply chain is one such innovative idea that is fast gaining attention in the industry. Today green supply chain is at the heart of the concept of sustainable development. This concept highly concerns about the environment. Eco-efficiency and remanufacturing processes are now important assets to achieve best practice (Srivastava, 2007). This concept is simply to produce more quality (environment friendly) output from less input. Reducing waste and pollution, and using less energy and material resources, are obviously good for the environment and evidently, are the best for supply chain because they cut the operational cost. Waste minimization is being considered as an important strategy towards attaining a green supply chain. Milk supply chain is more concerned with controlling the milk quality and supply fluctuations which are unique to this sector. Here, traditional supply chain is upgraded to highly effective value system that creates more value to all the partners in the supply chain. The Sri Lankan supply chain for milk and milk products is affected by wastage and poor handling. Wastage occurs due to presence of multiple points of handling. Contamination of milk can lead to huge economical losses. Contamination occurs at different levels: at farm level, during collection and storage, and at processing centers. Shortage of cold storage facilities and refrigerated transport equipments lead to inefficiencies in handling milk and milk products. Thus there is a compelling requirement for appropriate infrastructure facilities for temperature controlled warehouses, bowsers, wholesale and retail shops, etc. where storage and transportation activities are taking place. By practicing improved supply chain management practices, there will be a significant reduction in the wastages of milk and milk products which in turn will benefit both the farmers as well as the consumers by means of increased returns and decrease in prices respectively. Methodology In this project, firstly secondary data were collected to get an idea about the dairy industry in Sri Lanka. Secondary data were obtained from Ministry of Livestock Development, Central bank, Department of Census and Statistics, Department of Animal Production and Health Livestock statistics, Mahinda Chinthana, Sri Lanka customs, National cleaner production center (NCPC), publications of Central Environmental Authority and Sri Lanka Standard Code of hygienic practice for dairy industrie, etc. According to the secondary data obtained, there were no doubts about the huge potential for the expansion of the dairy industry in Nuwara Eliya district in the
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view of its favoured climate, labour, green pastures and the demand for milk from the other districts. Therefore, this community based dairy industry has been planned to be established based on the Kotagala farm which is located closer to the IDB Industrial Estate in Kotagala. Further, there are possibilities to increase the capacity of the dairy unit as it has been planned to locate in an area where there are plenty of small scale dairy farmers who sell their milk through intermediary salesmen. Designed community based dairy factory is planned to carry out the processing operations based on GMPs. Arrangements have been made to ensure hygienic conditions in milk processing by considering each and every aspect of milk processing starting from establishment designing, equipment purchasing and maintenance, control of operations, cleaning and sanitation, personal hygiene, transportation, staff training, pest control, waste management up to packaging and labeling. In here, greening the dairy supply chain, starting from farm level to distribution of finished products was studied. It includes raw material extraction, transportation, manufacturing process, wholesaler or retailer and consumer. Raw materials and energy wastage, underutilization of resources, environmental impacts and public health risks associated with each step in dairy supply chain was identified using primary and secondary data. Primary data collection was carried out using Observational method, Staff interviews and the Check lists were used to identify pollutes and impact of each pollute. Secondary data were obtained from the publications of Central Environmental Authority, National cleaner production center (NCPC) and research publications were used to identify the types of effluents discharged by the dairies and to find out the ways of maximizing overall environmental performance of a community based dairy industry. Overall cost effectiveness was determined by using a feasibility study. Results and discussion In manufacturing process, the processing factories can apply green concepts by several methods to reduce energy and resource consumption. This is where the reuse and recycling have to be concerned. Some practices include reducing energy consumption, recycling and reuse, using biodegradable and non-toxic materials, minimizing harmful emissions and minimizing or eliminating waste. The dairy farm produces the milk and it is collected by a collecting point thereafter bowser which delivers it to the dairy factory. At the dairy factory the milk is processed into a variety of dairy products and packaged for the consumer. After that they are delivered to the retail shops where the products are displayed for consumers on a refrigerator shelf or in a cold room for selling. A dairy item purchased by a consumer is transported to the household and stored in the refrigerator before the final consumption. Each of these activities in the milk chain causes environmental impact. When consider the dairy chain waste products, those may occur as liquid waste, solid material, volatile compounds or gasses that are discharged into the air. If this waste is not managed properly, it will directly affect the environment. Water pollution, soil pollution, air pollution are major environmental impacts in the supply chain. The major environmental impact associated with dairy supply chain is the pollution of surface and ground water. This would contribute to the organic load of the effluent stream (milk solids, detergents, sanitizers and milk wastes). Discharge of waste water to surface waters affects the water quality in three ways: 1.The discharge of biodegradable organic compounds (BOCs) may cause a strong reduction of the amount of dissolved oxygen, which in turn may lead to reduced levels of activity or even death of aquatic life. 2. Macro-nutrients (N, P) may cause eutrophication of the receiving water bodies. 3. Agro-
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industrial effluents may contain compounds that are directly toxic to aquatic life (e.g. un-ionized ammonia). Milk losses occur as a result of overflowing, spillage, foaming, leakage, and also during processing and cleaning. The majority of milk wastage occurs due to residual milk in storage tanks, pasteurizer, homogenizer, pipelines etc, and improper handling /usage of milk & other raw materials in dairy factory leads to resource waste. Refrigerant loss in milk cooling was identified as the major way of energy loss in dairy supply chain. Poor personal hygiene in the factory level is responsible for the food poisoning outbreaks which is the major public health significance in dairy supply chain. Financial evaluation of this project indicates a return on investment (ROI) of 57.14%. This Community based dairy factory will create employment directly to 21 persons and indirect employment to around 15 -20 farmers. Conclusions Application of green supply chain management approaches in dairy industry helps to achieve environmental friendly operations through resource conservation and waste management whilst achieving economic benefits. At the same time, this would help to ensure the quality and safety of the final products. There is a good market potential for the dairy products in Sri Lanka. This opportunity can be utilized to set up a profitable community based dairy factory in Kotagala area. References Mohanty, R.P., Deshmukh, S.G., 2008. Essentials of Supply Chain Management. Towards a green supply chain, 274-280.
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Development of Egg Less Cake Incorporating Yoghurt

T.B. Karunarathna, A.M.N.L. Abesinghe, D.C. Mudannayake Uva Wellassa University, Badulla, Sri Lanka and D.M.J.N. Danasekara Lucky Lanka Milk Processing Company, Karagoda, Uyangoda, Kamburupitiya, Sri Lanaka Introduction Cake is a product obtained from a batter containing wheat flour, sugar and eggs or wheat flour, shortening, sugar, eggs and other ingredients of requisite mass, put into trays and baked in an oven at suitable temperature for a suitable time (Sri Lankan Standards, 1995). The most primitive people in the world began making cakes shortly after they discovered wheat flour. They were described as flour-based sweet foods as opposed to the description of breads, which were just flour-based foods without sweetening. Bread and cake were somewhat interchangeable words with the term "cake" being used for smaller breads. Cakes are five types according to the Sri Lankan Standard specifications; cakes, butter cakes which contains wheat flour, butter, sugar and eggs without filling or any coating, fruit cakes that contain wheat flour, shortening, sugar, eggs, fruits (dry or preserved) and other ingredients, sponge cakes that contain wheat flour, sugar and eggs and cake with icing which are sandwiched and/or coated either with dairy or non dairy cream, jam, jelly, marshmellow, caramel, dried fruits or any other suitable mixture. The term yoghurt can be defined as A fermented milk product obtained from coagulation of milk specified as, cow or buffalo milk, standardized milk, skim milk or partially skimmed milk and reconstituted milk and concentrated milk by the agency of organisms of types Streptococcus thermophillus, Lactobacillus bulgaricus, Lactobacillus acidophilus may be present (Sri Lankan Standards, 1989). Yoghurt can be broadly categorized in to two types based on method of production, set yoghurt and stirred yoghurt. There are three types of set yoghurt in the local market; normal yoghurt, low- fat yoghurt and non-fat yoghurt. Stirred yoghurt can be found as plain, fruit or flavoured yoghurts (Tamime and Robinson, 2007). This study was carried out to develop an eggless cake for vegetarians by replacing eggs with yoghurt which is rejected just before the expiry date and thereby add value to yoghurts and cakes through product diversification. Methodology Cake was prepared using wheat flour, sugar, butter and baking powder. Three cake samples were prepared by changing wheat flour percentage as 15% (w/w), 20% (w/w) and 25% (w/w). First, wheat flour and baking powder were mixed in a bowl until well combined. Then, sugar and butter were measured and added in to the mixture at the room temperate and mixture was beaten well for 5 minutes using an electric beater to prepare cake batter. Batter was transferred to the greased trays and baked for 45 minutes at 180 oC. Baked cake was wrapped with polythene and stored under room temperature. Optimum wheat flour percentage was selected by a sensory evaluation using 30 untrained panelists at day 01. Wheat flour percentage Selected from above was used to develop yoghurt incorporated cake. There were three yoghurt incorporated cake samples by changing yoghurt percentage as 20% (w/w), 25% (w/w) and 30% (w/w). 110
First, wheat flour and baking powder were mixed in a bowl until combined well. Then, sugar and butter were measured and added in to the mixture at the room temperate and mixture was beaten well for 5 minutes using an electric beater to prepare cake batter. Preservatives were added in maximum level to the batter to increase the shelf life. Finally, yoghurt (4% of fat and 8.5% of SNF and vanilla were added to the batter and mixed well. Batter was transferred to the greased trays and baked for 45 minutes at 180 o C. Optimum yoghurt percentage was selected by a sensory evaluation using 30 untrained panelists at day 01. Selected cake sample was wrapped with polythene and stored under room temperature for further analysis. Sensory evaluations were carried out using a 7-point hedonic scale (7like extremely, 1 - Dislike very much) and sensory attributes such as appearance, colour, smell, taste and overall acceptability were assessed. Shelf life determination for selected cake sample was done by analyzing pH, yeast and mould count, total plate count, coliform and Escherichia coli at one week interval for 60 days of storage. Protein content, fat content and moisture content were analyzed by Kjeldhal method, Soxhelt method and oven dry method respectively as described in AOAC (2002). Data obtained from sensory evaluation were analyzed by Friedman rank test in MINITAB 14 software package. Results and discussion According to the sensory evaluation results of the cake sample with different wheat flour percentages, 20% (w/w) wheat flour incorporated cake had highest overall acceptability among three samples. Therefore, with the 20% (w/w) wheat flour, it gives best sensory characters to the cake. Furthermore, cake sample with 20% wheat flour and 30% yoghurt has highest preference with respect to overall acceptability. Therefore, 20% wheat flour and 30% yoghurt were selected as the best percentages to develop eggless cake. They can be stored for 60 days under room temperature without any quality deterioration. Similar to that, there is no growth of coliform, E. coli, yeast and moulds observed. The moisture content of the product was 26.28% and it fulfills the requirements of the Sri Lankan Standards for cakes with respect to moisture content. Considering the nutritional quality of the product, it has 6.13% protein and 7.94% fat. Fat content of this eggless cake is lower than the fat content of cakes with eggs and therefore it is more suitable for the people having high cholesterol levels. Conclusions Eggless cake can be developed using 20% wheat flour and 30% yoghurt with good sensory attributes. It can be stored for 60 days without any quality deterioration. Similarly, the fat content of this cake was lower than the cakes produced with eggs and therefore, more suitable for people who conscious on dietary fat contents. References AOAC. 2002. Official Method of Analysis of AOAC International, 20th ed: Association of Official Analytical Chemists, Maryland, USA. Sri Lanka Standards 1074 1989. Specification for cakes. Sri Lanka Standards Institution, Colombo. Tamime, A. Y. and R. K. Robinson 2007. Tamime and Robinsons Yoghurt Science and Technology, Woodhead Publishing Limited, Cambridge, England.
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Evaluation of Different Culture Types and Development of a Set Yoghurt With Cost Optimized Culture Option
S.A.V. Padmaraja , A.M.N.L. Abesinghe , D.C. Mudannayake , Animal Science Degree Programme. Uva Wellassa University. Badulla. Sri Lanka and M.N.P. Perera Fonterra Brands Lanka (Pvt) Ltd., Biyagama. Sri Lanka Introduction Over the last decade yoghurt and its preparations have developed into one of the most well-accepted and consumed acidified products. Mild acidic tastes, good digestibility, variations in taste and high dietetic value as well as stable quality have contributed to this growth. The starter culture is a critical factor in the production of set yoghurts it influences the organoleptic properties of the set yoghurt. A few studies have been conducted on evaluating the potential of using different culture types for yoghurt production. Kumari (2001) reported about the selection of a starter culture to improve the texture of plain set yoghurt at reduced total solid levels. Wijesinghe (1997) tested production of yoghurts using different ratios of Streptococcus thermophilllus and Lactobacillus bulgaricus and found that the best ratio of Streptococcus thermophilllus and Lactobacillus bulgaricus is 1:1. This study was carried out in one of the dairy factory in Sri Lanka where probiotic yoghurts are produced using two imported yoghurt starter culture types as base culture and probiotic culture. Base culture includes S. thermophilus , L. bulgaricus and Bifidobacterium species. From these three species, first two are considered as authentic yoghurt starter bacteria whereas the other is a probiotic bacterium. Probiotic culture includes Bifidobacterium lactis. The viable bacteria count in probiotic yoghurts at the end of shelf life is 106 cfu mL-1 However, it was found that the probiotic bacteria in base culture do not contribute much to maintain the viable probiotic bacterial population in set yoghurt. Therefore, the main objective of this study was to select a suitable nonprobiotic base culture for the existing set yoghurt without changing its organoleptic properties and thereby optimize the cost of set yoghurt production by selecting suitable non-probiotic base culture. Methodology Set yoghurts were prepared according to the standard procedure. The production process of set yoghurt includes yoghurt mix preparation, culture solution and skim milk solution preparation, activation of cultures, inoculation of activated cultures, incubation of yoghurt cups and transferring to the refrigerator. Selection of suitable base culture was done through trial and error method. Trials were conducted to compare new products with current product formulation. Two different multiple species non-probiotic cultures were used (A and B). The yoghurts prepared with non-probiotic base cultures were compared with the existing product for its organoleptic characteristics. Culture type C was the existing probiotic base culture. Both A and B included Streptococcus thermophilus, Lactobacillus delbrueckii subsp. Bulgaricus whereas C included Streptococcus thermophilus, Lactobacillus delbrueckii subsp. Bulgaricus and Bifidobacterium species. Completely Randomized Design (CRD) comprising three 112
treatments in ten replicates was used as the experimental design. Treatments were; treatment 1 base culture A + probiotic culture, treatment 2 base culture C + probiotic culture (existing set yoghurt) and treatment 3 base culture type A + B + probiotic culture. Treatment 3 (existing set yoghurt) was used as the control. Parametric data analysis was done using ANOVA for significance under =0.05 level using Minitab 14 statistical software package. Non parametric data analysis was done by Friedman non - parametric test using Minitab 14 statistical software package. In this analysis, 95% confidence interval was considered. The sensory evaluation was carried out with 20 semi trained panelists using nine point hedonic scale to assess sensory attributes of appearance, flavour, texture and overall acceptability. Three sensory evaluations were carried out to determine significant differences between sensory attributes of selected set yoghurt and existing set yoghurt. Shelf life determination was done by analyzing titratable acidity, pH, yeasts and moulds, coliforms at five days intervals for 35 days compared with existing set yoghurt (control sample). Incubation time was measured in final product. Probiotic count was measured during 14, 21, 28 and 34 days under refrigerated storage. Cost analysis for starter cultures was done and compared with existing set yoghurt production. Results and discussion Treatment 1, base culture type A and probiotic culture added set yoghurt was rejected after five repeated trials. According to the preliminary studies of culture types, treatment 3 (combination of culture type A and B with probiotic culture) was selected as the best non-probiotic culture for further analysis. There were no significant difference (p>0.05) between sensory attributes of appearance, flavour, texture and overall acceptability of non-probiotic set yoghurt and the control set yoghurt. Results revealed that total coliform, yeast and mould counts, pH and titratable acidity were in conformity to the Sri Lanka Standards limits. Oraganoleptic characteristics and incubation time of selected set yoghurt were similar to the control. The probiotic count during the refrigerated storage is higher than 106 cfu mL-1 in both yoghurts. The cost optimized by 5 cents per set yoghurt using selected culture and it can be stored at 4 C up to 35 days without reducing the viable probiotic counts than standards. Treatment 1 was rejected because of ropiness of texture compared to the control. Treatment 3 was selected after three repeated trials and three sensory evaluations because there were no difference (p>0.05) between sensory attributes of appearance, flavour, texture and overall acceptability of non-probiotic set yoghurt and the control set yoghurt. Coliform count was zero in both yoghurts during 35 days of shelf life period. This may be due to minimum level of contaminations, strictly maintained hygienic practices and addition of preservatives to the set yoghurts. The survival of Coliform bacteria is very low in lactic acid medium. There were zero counts of yeast and mould, because addition of preservatives to the yoghurt mix inhibited their growth. Similarly, the pasteurization of yoghurt mix helped to inhibit the growth of these microorganisms. There was no significant difference (p>0.05) of pH in set yoghurts during incubation period and the refrigerated storage period. The titratable acidity of selected set yoghurt was within the range of 0.8 - 1.25% lactic acid (w/w) and it complies to the Sri Lanka Standard specifications. Therefore, it revealed that, non-probiotic base culture has not contributed to the post acidification of yoghurt compared to the control. The viable probiotic counts of the probiotic yoghurt were within the standards (106 cfu mL-1) and it
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indicates that the elimination of probiotic bacteria from the base culture did not affect the viable probiotic count during its shelf life. The cost optimized by 5 cents per set yoghurt due to the usage of non-probiotic base culture and it reduced the cost of production of set yoghurt by Rs. 3,000,00 per month. Conclusions It can be concluded that the combination of non- probiotic base culture A and B with probiotic culture was selected as the cost optimized culture option for the set yoghurt production. References Kumari, A.A.T.T. 2001. Selection of a starter culture to improve plain set yoghurt texture at reduced total solid levels, B.Sc. Dissertation. University of Peradeniya, Sri Lanka. Wijesinghe, S.A. 1997. Producing yoghurt using different ratios of Streptococcus thermophillus and Lactobacillus bulgaricus, B.Sc. Dissertation, University of Peradeniya, Sri Lanka.
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Investigation of Genetic Variation in Bmp4 Gene in Local Indigenous and Jamnapari Crossbred Goats in Damana Veterinary Service Division Sri Lanka
H.R. Wijesena , P.B.A.I.K. Bulumulla Uva Wellassa University, Badulla, Sri Lanka L.G.S. Lokugalappatti and H.B.S. Ariyarathne Department of Basic Veterinary Sciences, Faculty of Veterinary Medicine and Animal Science, University of Peradeniya Introduction Small ruminants, such as goats (Capra hircus), constitute an important livestock resource in most countries and are essential for the livelihood of many farmers (Baker et al., 2003). Application of molecular genetics approaches for the genetic progress of quantitative economic traits such as growth and reproduction in goats is an effective way of increasing their production as these methods could lead to finding of genetic markers useful for improved selection. Molecular genetics approaches have been used in the world for goat production in the recent past, and these strategies are yet to be established in Sri Lanka since they require high knowledge and capital investments. Therefore, this study was conducted as a preliminary step for the application of molecular genetics approaches in selection of goats for improved production in Sri Lanka. Single Stranded confirmation Polymorphism (SSCP) analysis is one such powerful genetic screening method to identify the sequence variation in Polymerase Chain Reaction (PCR) amplified products. In the present study, we investigated the PCR-SSCP genetic variation in the intron 2 of Bone Morphogenetic Protein 4 (BMP4) gene, which plays a major role in growth and reproduction. The study was focused on Local types (LT) and Jamnapari crossbred (JC) goats in Damana Veterinary Service (VS) division in the Ampara district of Sri Lanka. Materials and methods Venous jugular blood samples were collected from total of 72, LT (18) and JC (54) goats in 10 farms from the Damana VS division of the Ampara district. Genomic DNA was extracted using QIAamp DNA Blood Mini Kit and target region was amplified using previously published (Fang et al. 2009) forward (5CTGGGGAAATGTTTGGTA 3) and reverse (5-GCTAAGAGTTG GGTGATGAG 3) primers. The PCR cycling protocol was 3 min at 94C, 40 cycles of 94C for 45 sec, 49C annealing for 45 sec, 72C for 1 min, with a final extension at 72C for 30 min. SSCP method was used to investigate different conformation pattern in the BMP4 gene with single stranded fragment movements. Aliquots of 3.5 l PCR products were mixed with equal volume of loading solution (95% formamide deionized, 25 mM EDTA, 0.025% xylene-cyanole and 0.025% bromophenol blue), heated for 4 min at 100C and chilled in ice immediately. Denatured DNA was subjected to 12% PAGE (polyacrylamide gel electrophoresis) in 1X TBE buffer at a constant temperature of 4C. Amplified fragments were separated initially at 300V for 5 min followed by 130V, 5W and 6mA current for 18 hours. At the end of the electrophoresis gels were stained with 0.1% silver nitrate. The DNA banding patterns were observed, recorded and photographed with GeneSys gel documentation system (Syngene). Frequencies of each conformational pattern were calculated for both LT and JC animals separately.
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Results Polymorphisms were detected in intron 2 region of BMP4 gene in both LT and JC goats in Damana VS division. Altogether three different conformation patterns were observed and the three patterns were designated as A, B and C (figure 1).
Figure 1: The electrophoresis patterns of PCR-SSCP for intron 2 of goat BMP4 gene. English letter corresponds to the conformational pattern. First lane indicates the 1kbp size standard. The area demarcated by line represents the schematic view of each conformational pattern. All the three conformational patterns (A, B and C) were found in both Local Indigenous and Jamnapari crossbred animals. Pattern A was predominantly found in both local indeginous (66.67%) and Jamnapari crosses (72.22%) in all the ten farms. Next to pattern A, pattern B has the highest frequency, whereas pattern C was found to be at lowest frequency in both breeds (Table 1). Table 1: Frequency distribution of each conformational pattern resulted in PCR-SSCP of intron 2 of goat BMP4 gene Breed Frequency of Conformational Patterns Pattern A Pattern B Pattern C 66.67% 72.22% 70.83% 27.78% 18.52% 20.833% 5.56% 9.26% 8.33%
Locals Jamnapari Cross Total frequency of each pattern
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Discussion This study was carried out as a part of a detailed study aiming at genetic characterization of indigenous goats in Sri Lanka for economically important traits, mainly growth and reproduction using molecular markers. Several studies have been conducted on BMP4 gene in different species including human, mice and bovine worldwide (Fang et al., 2008), and such previous studies conducted in China (Fang et al., 2009 and Chu et al., 2010), has described three conformational patterns (genotypes) with two alleles for the same gene fragment in three goat breeds for growth and reproduction traits. Similarly gene sequencing of the three observed conformational patterns to identify the genotypes and the alleles present is being pursued in order to determine whether any of the three conformation patterns observed in the study are corresponding to the previously reported polymorphism. According to data obtain from the study, patterns A and B were shown by both local and Jamnapari crossbred goats. But pattern C was mainly shown by Jamnapari crosses, except one local goat in farm 4. But pattern C was not identified in farm 10 where only local goats are being reared. Since pattern C cannot be seen in animals from farm 10 and as it was mainly seen in Jamnapari crosses, we can predict that this pattern may be inheriting from Jamnapari goats rather than locals. Same time we can predict that the local goat that showed pattern C in farm 4, may be a Jamnapari cross where Jamnapari characteristics are not well expressed. But to obtain accurate results and to prove these predictions, we need to sequence the three patterns. However, these results and conclusions should be considered as preliminary ones, and further investigations will be essential for detecting the polymorphism of this gene in all part of the country. Conclusions Based on results obtained, it can be concluded that goats in the study area are polymorphic for the intron 2 of BMP4 gene and possess at least three genotypes, alleles or allelic groups. However these conclusions were preliminary and sequence variation based on Single Nucleotide Polymorphisms (SNPs) of these populations is yet to be analyzed. References Baker, R.L., S.Nagda, S.L. Rodriguez-Zas, B.R. Southey, J.O. Audho, E.O. Aduda and W, Thorpe 2003. Resistance and resilience to gastro-intestinal nematode parasites and relationships with productivity of Red Maasai, Dorper and Red Maasai Dorper crossbred lambs in the sub-humid tropics. British Society of Animal Science. 76:119-136. Chu, M. X., L. Lu, T. Feng, R. Di, G.L. Cao, P.Q.Wang, L. Fang, Y.H. Ma and K. Li 2010. Polymorphism of bone morphogenetic protein 4 gene and its relationship with litter size of Jining Grey goats. Mol. Biol. Report. Xingtang Fang, Haixia Xu, Chunlei Zhang, Hong Chen, Xiucai Hu, Xueyuan Gao, Chuanwen Gu and Wangping Yue 2009. Polymorphism in BMP4 gene and its association with growth. Molecular Biology and Reprodcution 36:13391344.
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Evaluation of the Effect Of Artificial Insemination (Ai) on Hatchability in Indigenous Chicken at Central Poultry Research Station, Karandagolla
N.D. Andaraweera , N.M.N. Nambapana Uva Wellassa Univesity Badulla, Sri Lanka and G.A. Gunawardana Institute of Veterinary Research, Gannoruwa,Peradeniya,Sri Lanka Introduction The poultry farming is considered as one of the main livestock sector in Sri Lanka where indigenous chicken farming provides a promising house hold income for people in rural areas of Sri Lanka. (Buvanendran,1976).They can get high quality and quantity of day old chicks and eggs for daily consumption by rearing indigenous chicken.(Kushanthi et al., 2003) Introduction of Artificial Insemination (AI) program for indigenous chicken can be used for selection, upgrading and development of several suitable indigenous chickens for back yard poultry farming in Sri Lanka (De Silva, 1964). This experiment was carried out at the Central Poultry Research Station, Karandagolla under the supervision of staff of Veterinary Research Institute, Gannoruwa. The study was conducted to evaluate the effect of the Artificial Insemination on hatchability in indigenous chicken and to supply increased number of indigenous chicks to farmers by improving hatchability through AI which is the best method for breeding while increasing hatchability. . Methodology The study was conducted between 6th of March 2011 to 6th of August 2011,at the Central Poultry Research Station, Karandagolla, Kundasale located in the mid country intermediate zone in Sri Lanka under the administration of Veterinary Research Institute. Two samples and two replicates were prepared by conducting Artificial Insemination for one group and conducting Natural Breeding for one group. For Artificial Insemination program 6 male birds were used for semen collection and 40 female birds used for receiving semen. For natural breeding program 4 male birds and 40 female birds were used for better mating performance. All the other management factors feeding, water was given constantly for both groups. After eggs were collected those were incubated in the same conditions. After 18th day the candling data were collected and after 21st day hatching data were collected. Incubator parameters were monitored daily. egg breakout analysis were done after every candling and hatching. Finally, data were analyzed using SPSS 18 version statistical package. Paired t test was performed for comparisons of the data considering the difference between the groups at 5% level of significance. Results and discussion The mean fertility in the Artificial Insemination Program was significantly lower (25.26) than the mean fertility in Natural breeding program (34.62) (P<0.05).
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Fertility
40 35 30 25 20 15 10 5 0 1 2 3 weeks 4 5
Fertility %
Natural breeding AI program
The mean Flock performance hatchability (27.60) in Natural breeding program is higher than the mean. Flock performance hatchability (23.44) in Artificial Insemination program. There was no significant difference P>0.05 between Flock performance hatchability in Natural Breeding with the flock performance hatchability in Artificial Insemination in indigenous chicken.
Flock performance hatchability

40 35
Hatchability %
30 25 20 15 10 5 0 1 2 3 weeks 4 5 Natural Breeding
.
The mean of the sellable hatchability in the Artificial Insemination Program is 93.74, while the mean of the sellable hatchability in Natural breeding program is 81.84. There was a significant difference (P<0.05) between sellable hatchability in Natural Breeding with the sellable hatchability in Artificial Insemination in indigenous chicken.
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Selleble Hatchability
120 100
Hatchability %
Natural Breeding Aritificial Inseminati on
80 60 40 20 0 1 2 3 weeks 4 5
The study revealed that the Natural breeding Program is better than artificial insemination program for breeding of indigenous chicken. Further studies are needed to confirm these findings, since Artificial Insemination improves hygienic condition and also can be used for breeding indigenous chicken in Sri Lanka. References Buvanendran, V. 1976. Studies on hatchability of duck eggs, The Ceylon Veterinary Journal, 15(01):26-32, De Silva, P.L.G. 1964. Effect of female genotype on fertility and hatchability in chickens. The Ceylon veterinary journal, 12 (1 and 2). Kushanthi S.K.C., C.M.B. Dematawewa and D.V.S.de S. Gamage 2003. Evaluation of semen characteristics and fertility in a ecotypes of indigenous chicken , 11th Annual student research session, Department of Animal science, Faculty of Agriculture, University of Peradeniya. Acknowledgment We express our heartfelt gratitude to all the staff members in Central Poultry Research Station, Karandagolla and Staff members of Animal Husbandry school, Karandagolla.
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Study on Effect of Ginger Incorporated Broiler Feed on Body Weight Gain, Feed Conversion Ratio and Feed Intake of Broiler Chicken
R.M.D.S. Rathnayake Uva Wellassa Univesity, Badulla, Sri Lanka G.A.P. Ganegoda CIC Feed (Pvt)Ltd, P.O.Box 2 ,No.252,Kurunduwatte Road, Ekala,Sri Lanka and N. M.N. Nambapana Uva Wellassa Univesity, Badulla, Sri Lanka Introduction Modern intensive poultry production has achieved phenomenal gains in the efficient and economical production of high quality and safe chicken meat, eggs and poultry bioproducts (George, 1996). At the same time as making gains in production and efficiency, the industry had to maximize the health and well-being of the birds and minimize the impact of the industry on the environment. The use of feed additives has been an important part of achieving this success. Many additives have been a normal part of diets for animals and humans. It is only recently that we have come to recognize and understand their importance in achieving high production and efficiency, maintaining health and wellbeing, improving product quality.(Windisch et al., 2008) However, the feed additives have negative impacts on the consumers due to their residues which mostly remain in the broiler products. Thus, it is important to explore the potential of natural feed additives to replace the chemical ones.(Scott,2004) Probiotic and medicinal plants as natural feed additives are recently used in poultry diet to enhance the performance and the immune response of birds. One of the natural feed additives is ginger. Ginger contains bioactive substance such as oleoresin and ginger which give effect to optimize the body organ. Ginger also contains vitamins and minerals as the peculiar plant. Ginger extracts function as the anti inflammation and anti bacterial. Ginger is one of natural plants which can be used as phytobiotic to improve broilers performance. The major component of ginger is Zingiberen and Zingerol that can stimulate the digestive systems by controlling the digestive pH and the activity of digestive enzyme and the microbial activity. Ginger extracts could immune the gastric and improve poultry appetite. (Achyad et al., 2000) Studies show that the ginger spice has two types of digestive enzymes; one is protease enzyme that is used to break apart the protein and lipase enzyme that is used to break apart the fat. Both enzymes improve nutrients digestion and absorption by animals. Ginger is also as bacteria static that reduce pathogenic microorganisms in the digestive tracts. The gingerol also protect the liver on it activity, especially on hold the toxic of carbon tetrachloride. The ginger works as vaccination by stimulate an organ of bursafebrisius to make an antibody of viral attack such as ND. Ginger as a natural material is good as additive because it has no residual that threat the body organ and safe for the consumers health. (Peter et al., 1999) Methodology Broiler starter and finisher feed were supplied by CIC feed (Pvt) Ltd, and these feeds were taken as basal diets. Ginger was taken as feed supplement with basal diet. Raw
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ginger was brought in the same place to avoid composition variation. Ginger flour was prepared by washing the ginger under water and then it was slashed and sun-dried for 12 days. Dry ginger was then ground to get ginger flour. The ginger flour was stored in a plastic container to avoid chemical and microbiological damages. Ninety nine day-old female Hubbard flex broiler chicks were used for feed trial they were divided into three treatment groups, consisting 33 birds in each. The initial body weights of the birds were almost similar in all three treatments and average weight was 0.0484 kg.The chicks were reared until day seven in electrical brooder which has been divided into three separate compartments. Each compartment consisted forty watts (40 W) bulbs to provide heat, and each group was provided with around 6360 cm2 floor spaces. Artificial light was provided until third day during the day time and night. Thereafter, the lights were switched off by considering the behavior pattern of the chicks and environmental conditions to avoid over heating during the day time,. On the day seven, each treatment was replicated. Each treatment consisted of three replicates, and each replicate consisted of eleven birds. The experimental poultry house for growing birds (for day 7 to 42) consisted of nine cages. The replicates were arranged randomly within the nine cages. Each cage was equipped with separate feeder and drinker. Each cage provided 13500 cm2 of floor space and the height of the cage was 90 cm. The chicken in each group were given different feed as treatments. The feed were G-0 (control feed without ginger), G-1.0, and G-2.0 which were control feed with 1.0, and 2.0% ginger, respectively. The feed amount was changed according to the mortality. Ginger supplemented diets were provided separately within treatment groups. Broiler starter ration was given up to 28th day and the finisher ration was started from the 26th day. Birds were provided ad libitum clean drinking water throughout the study except in vaccination protocol. Multi vitamin mixture (Vita light) was given with drinking water in first five days of the study and after vaccination. The birds were vaccinated with ND vaccine on 3rd, 14th day and Gumboro (Infectious Bursal Disease IBD) vaccine on 14th, 21st, 28th day. Mortality and reasons for deaths were recorded throughout the period of study. During the brooding period (day 1 to 7), daily group feed intakes were recorded and weekly live body weights were measured on day eight. Following replication, body weights and feed intakes were recorded on replicate basis. In each replicate, daily feed intakes were recorded and weekly body weights were recorded on 15th, 22nd, 29th, 36th, and 43rd day. Average body weight gain and feed conversion ratio (FCR) were calculated using above measurements. Each variable was analyzed using Completely Randomized Design (CRD). Data were analyzed according to the General Linear Model (GLM) of ANOVA incorporated in Minitab 14. Three ginger samples were taken from three lots of ginger powder to prepare composite sample for the analysis. The ginger sample was subjected to sieve analysis and proximate analysis (crude protein, crude fat, crude fiber, moisture, and total ash). Results and discussion Proximate analysis indicated that the feed contained moisture 15.83%, ash 4.9%, crude protein 16.27%, crude fat 6.79% and crude fiber 6.87%. Weekly body weight gain, feed intake and FCR of the treatments are given in Table 1.
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Table 1: Weekly body weight gain, feed intake, and FCR of Broiler birds Parameter Period (days) 1-7 8-14 15-21 22-28 29-35 36-42 1-7 8-14 15-21 22-28 29-35 36-42 1-7 8-14 15-21 22-28 29-35 36-42 Basal dietBD (Control) 152.73 363.28 796.57 1276.18 1660.75 1828.15 121.18 412.39 951.54 1705.11 2660.44 3744.47 0.60 1.00 1.13 1.29 1.56 1.99 BD + 1% Ginger 149.52 368.49 790.19 1251.52 1666.24 2021.06 114.51 402.17 923.92 1643.20 2558.04 3584.55 58.00 97.00 1.10 1.26 1.49 1.73 BD + 2% Ginger 152.91 372.14 753.88 1249.94 1679.87 2099.62 114.30 400.59 917.15 1633.27 2550.88 3586.88 0.57 0.95 1.15 1.26 1.48 1.67
Body Weight Gain (g)
Feed Intake (g)
FCR
The particle size of ginger used in the experiment varied between 16 m and 30 m. According to the Table 1 the body weight gain of the chicks fed with ginger supplement diet has started to enhance at 5th week and in the 6th week it has become significantly different from the control. According to the ANOVA analysis (Table 2), the weight gain also showed a significant difference (p<0.05) with 2% ginger supplement. Table 2: Results of ANOVA analysis for weight gain Source Treatment DF 2 Seq SS 117082 Adj SS 117082 Adj MS 58541 F 14.92 P 0.005
Table 1 shows that the control group had higher feed intake than the groups which were supplemented with ginger throughout the study. According to the ANOVA analysis (Table 2), the feed intake indicated a significant difference (p<0.05) with 2% ginger supplement.
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Table 3: Results of ANOVA analysis for total feed intake Source Treatment DF 2 Seq SS 50412 Adj SS 50412 Adj MS 25206 F 43.38 P 0.000
According to the Table 1 FCR was higher in control group than ginger supplemented group. The FCR of the chicks t fedwith diet supplemented with ginger has enhanced at the 6th week and showed a significant difference (p<0.05) with 2% ginger supplement (Table 3). Table 4: Result of ANOVA analysis for feed conversion ratio Source Treatment Conclusions Dietary supplementation of ginger powder into broiler feed improves the Body Weight Gain, and decreases Feed Intake and Feed Conversion Ratio in an effective manner. There was different between 1% and 2% ginger supplementation for the effect body weight gain, total feed intake and FCR. Based on the research, it can be concluded that adding ginger as the supplement in the ration of broiler up to 2.0% gave a good effect on feed intake, total body weight gain and feed conversion. So, using 2% ginger supplement as phytobiotic additives in broiler diets will give higher body weight gain by consuming lower feed within 42 days. There is a potential to add value to nationally available ginger through the broiler feed industry. However, further studies are needed to investigate new phytogenic feed additives with more available herbs and medicinal plants in Sri Lanka, and to investigate the effect of ginger on the meat quality parameters such as color, drip loss, pH, cholesterol level, microbiological analysis and strength of meat. References George, J.M.1996. Poultry Prooduction Technology.The Avian Publication Company.West concert 34-49. Windisch, W., K. Schedle, C. Plintzner and A. Kroismayr 2008. Use of phytogenic product as feed additives for swine and Poultry. J.Anim.sci. 86:140-148. Scott, T.A. 2004.Cyber extention:Defining a feed additive.(Accessed on 25.05.2011)<http://www.poultryhub.org/index.php/feed_additives> Achyad., D.E and R. Rasyidah 2000. Jahe (Ginger). Retrived from http://www.Asiamaya.com/jamu/isi/Jahezingiberoffinale. htm, October 11, 2009. Peter, K.V. and Kandiannan, K.1999. Ginger.Tropical Horticulture. 1. DF 2 Seq SS 0.176156 Adj SS 0.176156 Adj MS 0.088078 F 23.66 P 0.001
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Reduction of Stress of Female Broiler Breeders During Growing Period to Maintain the Uniformity Level by Changing Temperature and Stocking density
H.M.W.G.S.L. Kumara and N.M.N. Nambapana Uva Wellassa University, Badulla, Sri Lanka
Introduction: Although the birds have high genetic potential for faster growth rate, better feed conversion, and increased meat yield in their progeny, if there is no optimum environment conditions for growth, the genetic potential does not appear in the environment. Among the factor for the better performances and health of the poultry birds, temperature and stocking density are the critical factors for stress of the birds in the poultry houses (Rosales, 1994). Chicken, unlike most other animals, do not possess sweat glands to aid in heat loss. The chicken removes excess body heat by radiation from the skin surface through the air to another object, by conduction to cooler objects with which the bird is in contact (Doug Grieve, 1990). Caged birds are more susceptible to heat stress because they are unable to seek a cooler place and there is less conductive heat loss in cages. As the environmental temperature approaches the body temperature of the bird, 41 C (106 F), the efficiency of these heat loss mechanisms diminish. At this point the evaporation of water from the respiratory tract becomes the major heat loss mechanism of the birds (Brake, 1987).The term "stress" is commonly used to describe the detrimental effects of a variety of situations on the health and performance of poultry. After extended or repeated periods of stress, birds become fatigued and weak, so they often succumb to starvation and infectious diseases (Rosales, 1994). Since the past, problems in broiler breeders are caused by combinations of whole house temperature and stocking density. Maintaining a uniform flock during the growing period of the broiler breeders may facilitate higher egg production during breeding period. So the broiler breeder farmers have to pay their attention to maintain over 80% uniform flock with increasing uniformity during both growing and breeding period for maximum production. The present study, aims to find better combination of whole house temperature and stocking density of broiler breeders to maintain over 80% uniform flock with increasing uniformity during growing period. Methodology This study was carried out at the NEL Farm & Hatchery that has been established under Noorani Estate Limited Naththandiya. Total of four hundred and ninety five Hubbard F15 broiler breeder hens were used from three weeks of age to eighteen weeks of age in this study. Forty five birds were divided in to nine replicates of five birds each as the control and remaining four hundred and fifty birds were divided in to nine group under temperatures of 27 29 C, 29 31 C and 31 33 C with stocking densities of 1 bird/ 1.8 ft2, 1 bird/ 2.0 ft2 and 1 bird/ 2.2 ft2. After two weeks of successful brooding period, the birds were subjected to above conditions from the beginning of third week up to the end of eighteenth week. The above temperature conditions (27 29 C, 29 31 C and 31 33 C) were maintained by using three grower cages at the grower farm of NEL Farm & hatchery. Each cage was partitioned in to three pens according to the above stocking densities. Cages were numbered as given in Table 1.
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Table 1: Conditions of the cages Cage number Temperature and Stocking density conditions/Treatments 27-29 C and 1.8ft 27-29 C and 2ft 27-29 C and 2.2ft 29-31 C and 1.8ft 29-31 C and 2ft 29-31 C and 2.2ft 31-33 C and 1.8ft 31-33 C and 2ft 31-33 C and 2.2ft
1 2 3 4 5 6 7 8 9
Broiler starter was given during first two weeks and chick starter was given from third week to end of fourth week. Then grower feed was given from sixth week to eighteen week. Broiler starter was given as ad libitum and then chick starter and grower feed were given from second week to eighteen weeks of age as restricted feeding. After fourth week of age feed restriction was practiced and birds were fed six days for a week (6/1 feeding programme). Body weights of the birds were recorded weekly in each cage during experimental period and uniformity of the each pen was calculated with the help of above weights. BAT 1 Poultry Scale was used to weigh and calculate the collected data. After sixteenth weeks of age, averages from weekly uniformity levels, standard deviation and coefficient of variance for each pen were calculated. Collected data was analyzed by using two-way Analysis of Variance (ANOVA) to analyze the differences between treatment groups using SPSS General Linear Models procedures. Results and discussion Two hypotheses were built relevant to the above study as given below. H0 There is no significant difference between uniformity with temperature and stocking density combination H1 There is a significant difference between uniformity with temperature and stocking density combination Component Var (VAR00003) Var (Error) Estimate 0.000 0.035
VAR00001 Dependent variable/Uniformity VAR00002 (Error) Stocking Density VAR00003 Temperature
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Results of the study revealed that there is a significant difference between uniformity with temperature (P<0.05) and stocking density (P<0.05) combination. Therefore the combination of temperature and stocking density affected for highest uniformity level can be selected as the best combination.
Uniformit y
1 0 1
0.45 0.24 0.24 0.43 0.93 0.42 0.51 0.310.34 2 3 4 5 6 7 8 9 Treatments
Figure 1.Variation of uniformity in different treatments According to the Figure 1 treatment no. 5 has given highest uniformity level than the other eight treatments. Therefore it is the better combination of temperature and stocking density for the broiler breeders to gain better performance. In this graph (Cage under 29-31 0C), fluctuation of the uniformity is very low; it has shown somewhat increasing uniformity level comparatively other cages. According to the graph, uniformity is over 80% with the 29 31 0C temperature. Under 2.0 ft2 stocking density, fluctuation of uniformity is lowest than the other conditions. Therefore, the best combination to keep these breeder birds is 29 31 0C temperature with 2.0 ft2 stocking density. References Brake, J.T. 1987. Stress and modem poultry management. Animal Production Highlights. F.6offman-La Roche and Co., Ltd., 4002 Basle, Switzerland. Doug Grieve, D.V.M., 1990. Technical Bulletin, A publication of Hy- Line International. Rosales, A.G. 1994. Applied Pouluy Science, Managing Stress in Broiler Breeders.
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Study on Effect of Curry Leaves Supplementation with Broiler Feed on Growth Performance, Feed Intake and Feed Conversion Ratio of Broiler chicken
W.W.H.A. Sampath , N.M.N. Nambapana Uva Wellassa Univesity, Badulla, Sri Lanka and G.A.P Ganegoda CIC Feed (Pvt)Ltd, P.O.Box 2,No.252,Kurunduwatte Road, Ekala, Sri Lanka Introduction The broiler industry has developed all over the world during past few decades. When considering the production of 79.9 million broiler chicks in Sri Lanka in 2007, there is an increase in 2008, 2009 and 2010 years (Department of Animal Production and Health, 2010). The production of poultry meat and other poultry products have been drastically increased in Sri Lanka within last few years. The world broiler meat production in 2010 was 73 million metric tons (USDA-FAS, 2010). China, Brazil, European Union, Mexico are the main Broiler producers in the world (USDA-FAS, 2007). Poultry meat and other poultry products such as eggs, have a higher demand in Sri Lanka. When considering the consumption pattern of the meat in Sri Lanka, chicken meat (broiler meat) has the highest demand and broiler meat has been included in the gazette as an essential food item in Sri Lanka since 2007. The requirement of nutrients for broilers is higher than the other livestock animals. Proper nutrition and the better intensive management practices are essentials in poultry industry. Hence feed cost is major cost component in poultry industry and it is accounted up to 60%-70% of the total cost of production. The production of feed in 2009 for poultry in Sri Lanka was 454,000 Mt. However the feed price has increased after 2008 and the profit margin of the industry has gone down (Department of Animal Product and Health, 2009). To overcome this limitation in the industry, feed supplementation is done by the farmers/producers. The supplementation is done using low cost, available feed stuffs and without affecting the performance of birds and the quality of the meat. Performance of the animal can be increased by increasing the feed conversion by improving the internal environment modification. This can be achieved by inclusion of antibiotics into feed. Antibiotics are the chemicals those which antagonistic towards or destructive of life (The penguin encyclopedia of nutrition, 1985). Some of other feed ingredients are used to restrict or avoid the usage of antibiotic growth promoters (AGPs). Some of those are probiotics, prebiotics, synbiotics, enzymes, acidifiers, antioxidants, phytogenic additives and herbal extracts (Pauline, 2009). The usage of natural plant based materials improves the feed intake, feed digestibility, feed conversion efficiency, the quality of the meat and reduce mortality (Hathurusinghe, 2008). Natural herbal materials increase colour lipid oxidation and reduce gut microbial content (Cross et al., 2007). Essential oils, organic acids and phytogenic compounds enhance production of gastric secretions, stimulate blood circulation and reduce level of pathogenic bacteria (Buchaan et al., 2008)
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This study was done to investigate the effect of curry leaves incorporated broiler feed on growth performance and feed conversion ratio of broiler chicken under field condition in Sri Lanka. For the study, the dried, blended curry leaves supplementation was used. The study hypothesized that dietary supplementation of curry leaves has an ability to improve the health, performance and reduce the cost of production of broilers. Methodology For the experiment, 99 day old male Hubbard flex chicks were divided into three treatment groups having 33 birds per each group. There were three treatments as control group who were fed with basal diet, 1% and 2% curry leaf supplementation respectively with basal diet. Brooding was practiced in first seven days by dividing only as treatment groups. Then each treatment group was divided into three replicates randomly in day 8th to 42nd of age. Initial body weights, weekly body weight and daily feed intake were measured during the experimental period. Birds were provided ad libitum clean drinking water throughout the study except in vaccination protocol. Multi vitamin mixture (Vita light) was given with drinking water in first five days of the study and after vaccination. The birds were vaccinated with ND vaccine on 3rd, 14th day and Gumboro (Infectious Bursal Disease IBD) vaccine on 14th, 21st, 28th day. Mortality and reasons for deaths were recorded throughout the period of study. During the brooding period (day 1 to 7), daily group feed intakes were recorded and weekly live body weights were measured on day eight. Following replication, body weights and feed intakes were recorded on replicate basis. In each replicate, daily feed intakes were recorded and weekly body weights were recorded on 8th,15th, 22nd, 29th, 36th, and 42nd day. Average body weight gain and feed conversion ratio (FCR) were calculated using above measurements. Each variable was analyzed using Completely Randomized Design (CRD). Data were analyzed according to the General Linear Model (GLM) of ANOVA (Minitab 14). Three curry leaves samples were taken from three lots of curry leaf powder to prepare composite sample for the analysis. The curry leaves samples were subjected to sieve analysis and proximate analysis (crude protein, crude fat, crude fiber, moisture, and total ash). Results and discussion Proximate analysis of the curry leave supplemented diet consisted moisture 21.98%, ash 9.92%, crude protein 6.10%, crude fat 1.00% and crude fiber 3.70%. Results of weekly body weight gain, feed intake, FCR and Results of sieve analysis are given in Table 1 and Table 2 respectively.According to the sieve analysis 08 m and 16 m were the dominating particle size of curry leaf used in the experiment. According to results of experiment the body weight gain of chicks fed with curry leaves supplemented diets were higher (P<0.05) than the basal diet. Final body weight gain of the birds fed basal diet was lower (P<0.05) than that of other two supplementary groups. These findings of the study agree with the observations of Hathurusinghe (2008) that states dietary herbal compounds improve the body weight gain and final body weight of broilers The feed intake of chicks and FCR in curry leaves supplemented diets were lower (P<0.05) than that of basal diet but weight gains were high in curry leaves supplemented diets. There was no significant difference (P>0.05) between 1% and 2% curry leaves supplemented diets for weight gain, feed intake or FCR. 129
Table 1: Weekly body weight gain, feed intake, and FCR of broiler birds Parameter Body Weight Gain (g) Period (days) 1-7 8-14 15-21 22-28 29-35 36-42 1-7 8-14 15-21 22-28 29-35 36-42 1-7 8-14 15-21 22-28 29-35 36-42 Basal diet-BD (Control) 144.87 361.33 769.33 1293.80 1828.68 2028.71 125.26 414.48 955.14 1711.11 2672.38 3747.32 0.64 1.00 1.16 1.27 1.42 1.80 BD + 1% Curry leaves 159.61 381.22 788.66 1288.66 1726.57 2235.40 116.72 404.31 927.56 1659.20 2567.24 3589.94 0.55 0.93 1.10 1.24 1.44 1.57 BD + 2% Curry leaves 150.58 372.66 772.51 1292.00 1788.00 2247.31 116.88 402.29 921.65 1644.89 2558.74 3583.41 0.58 0.95 1.12 1.22 1.39 1.56
Feed Intake (g)
FCR
Table 2: Results of sieve analysis Mesh Number (m) 07 08 10 12 16 30 35 Bottom plate Conclusions Dietary supplement of curry leaves into broiler feed significantly improves the Body Weight Gain (BWG) and decreases Feed Intake (FI) and Feed Conversion Ratio (FCR) in an effective manner. The current study revealed that the curry leaves can replace dietary antibiotics and since there was no significant difference between 1% and 2% Curry leaves Powder (g) 02 10 05 02 07 02 01 03 Percentage 06.25 31.25 15.62 06.25 21.87 06.25 03.12 09.37
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curry leaves supplementation on the BWG, FI and FCR, 1% curry leaf supplementation is adequate to be used in broiler industry. The study showed that there is a potential to add value to nationally available curry leaf plant through the broiler feed industry. However, further studies are needed to investigate new phytogenic feed additives with more available herbals and medicinal plants in Sri Lanka. References Department of Animal Production and Health, Annual Report, 2009. Peradeniya, Kandy, Sri Lanka. Department of Animal Production and Health, Annual Report, 2010. Peradeniya, Kandy, Sri Lanka. USDA-FAS attach reports. 2010. Official statistics and results of Office Research, United States Department of Agriculture. USA. USDA-FAS attach reports. 2007. Official statistics and results of Office Research, United States Department of Agriculture. USA. The penguin encyclopedia of nutrition 1985. Penguin books Ltd, Harmondsworth, Middlesex, England. Pauline, G. 2009.Cyber extention:Antibiotics and the mode of action.(Accessed on 20.06.2011)Available at <http://ezinearticles.com/?Antibiotics-And-The-Mode Of-action and id =1193644> Hathurusinghe, H.D.K.C. 2008. Potential use of selected herbs and spices as alternatives to antibiotics in broilers. Final year research project, Department of animal science, Faculty of Agriculture, University of Peradeniya. Cross,D.E., R.M. Devin, K.Hillman and T. Acamovic 2007. The effect of herbes and their associated oils on performance ,dietary digestibility and gut microflora in chickens from 7 to 28 days of age. Brit. Poult. Sci. 48:496-506. Buchaan ,N.P., J.M. Hott, S.E. Cultip, A.L. Rack and A. Asmer 2008.The effect of a natural antibiotic alternative and a natural growth promoter feed additive on broiler performance and carcass quality. J. Appl. Poult,17:202-210.
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Performance Evaluation of Chicks, Obtained Through a Selective Breeding Programme to Introduce into Backyard Poultry Farming
S.M.C. Weerasinghe , N.M.N. Nambapana Uva Wellassa University, Badulla, Sri Lanka and G. Gunawardene Department of Animal Breeding, Veterinary Research Institute, Gannoruwa, Peradeniya, Sri Lanka Introduction Poultry production has increased rapidly and tremendously in the last two decades in Sri Lanka (Gamage et al., 1993). The Department of Animal Production and Health (DAPH) Government of Sri Lanka during the past decade through the Central Poultry Research Station (CPRS), Karandagolla, Kundasale has been distributing upgraded indigenous chicken among Backyard farmers. Breeding Indigenous cockerels with Black shaver commercial layer hens is the breeding program practiced presently (2011) at CPRS to upgrade the Indigenous chicken. Performance evaluations of resulting chicks obtained through a selective breeding of Black Shaver hens with Indigenous cockerels is the first step of the project. Program was carried out at CPRS, Karandagolla. Methodology Hundred thirty eight breeder birds at age of twelve months were randomly selected for the study. Twenty hens and three cockerels were included in each mating group. Three treatment mating groups; a. Black Shaver hens with indigenous cockerels, b. Black shaver hens with Black shaver Cockerels and c. Indigenous hens with indigenous cockerels were maintained in constant conditions. The latter two treatments were regarded as Control 1 and Control 2. A replicate was maintained for each mating group. Separately collected, numbered, cleaned and fumigated eggs were set into brooder once per week. Eggs were candled on 18th day and transferred into Hatcher machine. Chicks were taken out on 21st day. Wing band was given to each bird for identification. Data of eggs, birth weights, weekly body weights, average feed intake per day and mortality were recorded in treatments and two control groups including replicates. Data were analyzed using Microsoft office excel and SPSS 16.0 analytical software. Central tendency, Dispersion, One way Analysis of Variance tests were conducted for the collected data. Results and discussion According to the eggs data analysis results at the 5% level of significance, there were a significantly lower number of fertile eggs (p<0.01) in Indigenous group than treatment group eggs. There were significantly higher number of good chicks in black shaver group than treatment (p<0.01) and indigenous groups (p<0.01).
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Feed intake of treatment chicks is significantly lower (p<0.05) in treatment chicks than black shaver chicks. There were significantly higher mortality (p<0.01) of treatment chicks than black shaver chicks and significantly lower (p<0.01) mortality of treatment chicks than indigenous chicks. Conclusions Considering all the results of study, the birth weights and weekly body weights of treatment chicks were almost similar to that of black shaver and indigenous chickens. Feed intakes of treatment chicks were lower than black shaver chicks. Treatment / Resulting chicks of selectively bred group had the better performance than indigenous chicks when considering most of the evaluated factors. Black shaver chicks had the best performance out of all three groups. Overall performance of treatment chicks were in between the black shaver chicks and indigenous chicks. References Gamage D.V.S., M.G. Jeyaruban, G.S. Wijekoon and G.G. Podimenike 1993. Development of local commercial egg laying strains at Central Poultry Research Station (CPRS) at Karandagolla,Annual report 1993,Sri Lanka Veterinary research Institute,Gannoruwa,Peradeniya.
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Effect of Different Pasteurization Temperature-Time Combinations on Shelf Life of Raw Cream in Relation to its Microbiological, Chemical and Physical Properties
C.N.P. Gunawardana, D.C. Mudannayake Uva Wellassa University. Badulla, Sri Lanka and M.N.P. Perera Fonterra Brands Lanka (pvt) ltd. Biyagama. Sri Lanka Introduction Cream is a vital ingredient in manufacturing of many dairy products. Cream is a good substrate for microbial growth due to its high nutritional value. Generally, in dairy processing factories separated cream is held on a period of time prior to incorporation in to the dairy products. The spoilage of cream from separation till the production of dairy products has been a critical problem to producers. The treatments which are given and the conditions under which cream is held will have a direct effect on its keeping quality. Shelf life of raw cream currently produced as an ingredient for curd production at Fonterra Brands Lanka (Pvt) Ltd is estimated to be approximately four days at 4 C. Therefore a method that could be used to extend the keeping quality of raw cream beyond four days would be a helpful and economical to the industry. Pasteurization of raw cream after separation can be done to improve the keeping quality. As there are no regulations governing heat treatment of cream in Sri Lanka, the time/temperature combinations used vary widely in practice. This investigation was undertaken to determine the effective pasteurization temperature/time combinations to improve the keeping quality of cream. Methodology Raw cream (60% fat) sample was obtained from the cream separator (Serial no: 95078, Frautech, Italy) in the factory. Initially the fat content of the sample was measured according to the Gerber method described in IDF 152 A: 1997. 4 kg of cream sample was divided in to 1 kg of four samples separately. Each 1 kg of cream sample was transferred in to Duran bottles aseptically and three bottles were used for each of the temperature. The cream filled Duran bottles were held in four temperatures as 72 C (T1), 75 C (T2), 78 C (T3) and 81 C (T4) for 15 seconds in the water bath (model:Wb29, Memmert, Germany). A thermometer inserted (OAKION, serial number; 2347890754, England) cream filled Duran bottle was kept along with samples in each trial to check the accuracy of the pasteurization temperature. Each temp trial was replicated. A complete randomized design was used to ensure that each temperature and time held a same position in processing in each three replicate. After heat treatment samples were cooled by immediate immersion in running cold water at about 12 C for 10 minutes and then the samples were transferred aseptically in to cups. As the control sample other 1 kg of cream sample was used. All the cups were kept in the cold room which has a temperature of 4 1 C. Initially microbiological (Aerobic plate count, Yeast and Moulds, Coliform), chemical (pH, titratable acidity) and physical parameters (colour, texture, odour, appearance) of cream samples were measured by getting three cups from each sample. All above
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parameters were checked in 3 days interval for 30 days at temperature 4 oC. The pH and titratable acidity data were analyzed by ANOVA (Analysis of variance) and Duncan New Multiple Range Test (DNMRT) from the statistical software package SAS 9.0. APC, Yeast and Moulds, coliforms were tested in cream processing area and handling equipments as well. Results As there are no regulations governing heat treatment of cream in Sri Lanka, the microbiological limits set out by the Bureau of Indian standards (2006) were used in this study; APC < 107 CFUg-1 (Raw Cream) and < 6 104 CFUg-1 (Pasteurized Cream), Coliforms < 100 CFUg-1 (Raw Cream) and < 10 CFUg-1 (Pasteurized cream), Yeasts < 1000 CFUg-1 and Moulds < 10 CFUg-1 (Raw cream), Yeasts < 100 CFUg-1 and Moulds < 1 CFUg-1 (Pasteurized cream). APC of control sample exceeds its microbiological limit (The Bureau of Indian standards, 2006) after the 3rd day of refrigerated storage (4 oC). But APC of treatments T1, T2, T3 and T4 exceed the microbiological limit (The Bureau of Indian standards, 2006) after the 9th, 12th, 15th, 9th days respectively. T1 and T4 exceed microbiological limit at the same day (9th day) at refrigerated storage. On the 9th day APC of T1 (6.97 104 CFUg-1) was greater than T4 (6.84 104 CFUg-1). Yeasts and Moulds were not detected in any treatment sample during the whole period of storage at 4 oC. After the 3rd day Yeasts and Moulds were increased markedly in control sample. At the end of the storage (30 days) Yeast and Mould counts detected in the control sample were 2.8102 CFUg-1 and 2.91102 CFUg-1 respectively. Throughout the study coliforms were not detected in any of the cream samples. Initial pH of the treatments (T1, T2, T3, and T4) 6.74, 6.73, 6.75 and 6.72 were declined with time up to 5.13, 5.79, 5.83 and 5.62. Control sample had the lowest initial pH value (6.71) compared that of pasteurized samples. pH of the treatments T1, T2, T3 and T4 decreased markedly after 9th, 12th, 15th and 9th days. Similarly titratable acidity of the treatments T1, T2, T3 and T4 were increased markedly after 9th, 12th, 15th and 9th day respectively. Titratable acidity of control sample was rapidly increased after the 3rd day. Colour of the T4 sample was impaired (turned to yellow colour) after pasteurization compared to other treatments. In the refrigerated storage, all the cream samples were thickened and the gelation was occurred and also slight putrefactive odour was occurred. Mould growth on the surface was observed only in the control sample at 5th day of refrigerated storage. Discussion Adherence to relevant regulatory requirements, not allowing microbial counts to exceed regulatory limits is important in determining the shelf life of cream. In the T4 (81 oC) APC was increased significantly after 9th day compared to other treatments. This is an agreement with Robinson (1999) who stated that a higher temperatures than 80 C may impair cream quality, possibly through activation of bacterial spores.
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Yeast and Mould colonies were not observed in treatment samples, while mold growth was started after 5th day of control sample. These results are due to destruction of Yeasts and Moulds in cream by the pasteurization. Throughout the study coliforms were not detected in any of the cream samples. These observations can be explained by the results of microbiological evaluation of the cream separation environment and the cream handling equipments in the factory. Coliforms were not detected in the floor of the cream separation area, neither in containers used to store cream nor in the cream separator. The results indicated that a high level of hygiene is maintained throughout the cream separation and storage in the factory. There was a significant (P< 0.05) difference in pH of pasteurized and the control samples that is mainly due to pasteurization. According to DNMRT the higher mean value for pH was in the T3. After the 30 days of storage at 4 C titratable acidity was very high in the control sample (0.297) than the treatments (T1-0.179, T2-0.169, T3-0.157, T4-0.218). The increment of titratable acidity is a reflection of souring activity due to lactic acid produced by microorganisms. According to Hammer, 1948 acid production by S. lactis is dominated in raw cream stored in 4 C. The increase of titratable acidity of control samples during the refrigerated storage can be explained with this statement. There was a significant difference (P<0.05) in lactic acid development during refrigerated storage of control and pasteurized cream samples. This could be due to the fact that pasteurization destroys many of the lactic acid producing microorganisms. The yellow colour development in T4 was due to the high pasteurization temperature. According to the Robinson (1999) lipolytic enzymes produced by psychotropic bacteria can result from long refrigerated storage of pasteurized cream. Psychotroph-derived proteases may also cause spoilage involving thickening and gelation. According to the findings of Robinson (1999) the thickening was accompanied by a slight putrefactive odour. Conclusions Pasteurization of raw cream shows a higher shelf life than the raw cream in refrigerated storage (4 C). pH and titratable acidity of the treatment samples were significantly differ (P<0.05) compared to control sample. From mean values in DNMRT the highest pH and the lowest titratable acidity were in the sample that was pasteurized to 78 C for 15 s. According to physical parameters minimum changes during the refrigerated storage was observed in sample pasteurized to 78 C for 15 s. T1, T2, T3, T4 and control sample had shelf life of 9, 12, 15, 9 and 3 days respectively according to microbiological data (The Bureau of Indian standards, 2006). The highest shelf life was detected in the sample pasteurized to 78 C for 15 s. The overall conclusion is that, it is possible to extend the shelf life of the raw cream by pasteurization process beyond four days. The best temperature time combination is 78 C for 15 s. References Bureau of Indian standards specifications, 2006. Bureau of Indian standards, New Delhi Codex standards for cream for direct consumption, 2003. Codex Alimentarius Commission. Crossley. E.L. 1948. Bacteriological flora and keeping quality of pasteurized liquid cream. Journal of Dairy Research, 15:261-276 Robinson, R.K. 1999. Modern dairy technology, Aspen publishers, Maryland, 1:61-101. 136
Effects of Supplementation of Nitrogen through Urea Molasses Multinutrient Block (UMMB) on the Performance of Dairy Cows Fed with Good Quality Forage Based Diets While Using Rice Straw as Night Feeding
D.R. Jayawickrama , D.C. Mudannayake, D.K.D.D Jayasena Uva Wellassa University, Badulla, Sri Lanka and W.M.P.B Weerasinghe Veterinary Research Institute, Gannoruwa, Peradeniya, Sri Lanka Introduction In Sri Lanka, the dairy industry is not well developed but has huge potential for the development. Among the constraints faced by the dairy industry, poor nutrition status of the animals has been identified as a major obstacle for the development of dairy industry in Sri Lanka. In general, animals are fed with poor quality roughages and concentrate feeding is very limiting thus, animals genetic potential for the milk production has not been achieved in many cases. Poor quality roughages contain very little energy and protein, which is responsible for the lower production. Several methods have been reported in Sri Lanka to improve the nutritive value of low quality roughages. Among those, UMMB feeding is one of the easier methods. Hard solid blocks of UMMB provide readily available sources of energy and protein in the form of molasses and urea together with fiber and minerals (Saddul and Boodoo, 2001). Urea-molasses mineral block (UMMB) licks can improve the utilization of low quality roughages by satisfying the requirement of the rumen microorganisms, creating a better environment for the fermentation of fibrous material and increasing production of microbial protein and volatile fatty acids (Wongnen, 2007). Urea, after hydrolyzing into ammonia in the rumen, provides a nitrogen source for the rumen microflora for their microbial protein synthesis. Molasses is a source of readily fermentable energy (Wongnen, 2007), which assists the growth of rumen microorganisms. It has been shown that animal performance has improved tremendously after the introduction of UMMB under field conditions (Kunju, 1986). This improvement was attributed to supplementary and catalytic effects of UMMB, as UMMB promotes an optimal ammonia level for efficient microbial activity in the rumen (Kunju, 1986). Several researchers have previously reported on the use of UMMB licks for supplementing the crop residue-based diet of large and small ruminants (Leng, 1983; Sansoucy, 1995) but very few studies have been conducted on the use of UMMB with good quality forage-based diets. Results of one such study by Weerasinghe et al. (2010) to evaluate the effects of supplementation of nitrogen through UMMB on the performance of dairy cows fed with good quality forage based diets, highlighted that UMMB supplementation significantly increased milk yield and yields of milk fat, protein, and SNF and UMMB supplemented animals had a significantly higher body weight than those fed with control diets; it suggests that the improvement of production and performance could be due to improved digestibility of the basal diet. However, no information available on the use of straw as night feeding to replace the amount of grass supplied in the day time. Thus, the objective of this study was to
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evaluate the effects of supplementation of UMMB to dairy cows fed with good quality forage based diets while supplying rice straw as night feeding. Methodology Ten multifarous crossbred dairy cows in their early lactation were randomly allocated to two groups (Supplemented and control) based on their milk yield, breed, parity, body weight, milk fat and protein contents measured in previous three days before feeding the experimental diets. Both groups were fed with chopped CO3 (Pennisetum purpureum x Pennisetum americanum; hybrid Napier) ad-libitum and dairy cow concentrate feed 1 kg/day during the day time and rice straw (5 kg dry matter) was supplied as night feeding. In addition, the treatment group was supplemented with 300 g/day of crushed urea-molasses multinutrient block at three equal meals (100 g at a time) during the day. The composition of UMMB was similar to that described previously by Weerasinghe et al., (2004). Throughout the experiment, the cows were penned individually and had free access to water. Cows were milked twice daily at 0700 and 1600 h using a mobile milking machine. Milk yield was recorded and milk samples were collected once a week throughout the experimental period (5 weeks) for laboratory analysis. The contents of milk fat, protein, lactose, and solid non fat were measured using an Ultrasonic Portable Milk analyzer (LACTOSCAN, SA type). Feed samples (mineral block and rice straw) were analysed according to the methods described in A.O.A.C. (2000). In addition, straw intake and weight gain were measured. The data were analyzed as a randomized block design using Genstat (Discovery Edition). Results and discussion Supplementation of urea-molasses multinutrient block had no effect (P>0.05) on straw intake. It can be observed that an increase in dry matter intake (DMI) is significant generally in studies where the basal diet consists mainly of poor quality roughages either hay or straw (Badurdeen et al., 1989) and when the quality of the diet improved with the inclusion of concentrate, the effect diminished. In this study, even though night feeding is totally based on poor quality roughage (i.e. rice straw), the supplementation with UMMB had not affected straw intake. This could be due to feeding of concentrate and, good quality fodder grass (i.e. CO3) ad-libitumly during the day time. Therefore it can be suggested that those feed might have been adequate in providing optimum nutrition to the animals, thus the cows were not in the need of extra dry matter intake. Supplementation of urea-molasses multinutrient block had no effect (P>0.05) on milk yield. But average milk yield (mean value) had increased numerically in the treatment group compared with the control diet fed animals. Similarly, UMMB supplementation had no effect (P>0.05) on contents and yields of milk fat, protein, lactose and solid non fat. But all those values were numerically high in UMMB supplemented animals compared to the control group. In addition, milk urea nitrogen content and weight gain was not affected (P>0.05) by UMMB supplementation. Conclusions It can be concluded that nitrogen (in the form of urea) supplied through UMMB provided with good quality fodder grasses and dairy cow concentrates with provision of rice straw as night feeding has no effect on production performance of dairy cows. As basal diet consisted of good quality roughage source and sufficient amount of
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concentrate feed, nutrients provided through UMMB would not be required by the animals for their milk production. Even though not significant, a numerical increment of milk production and quality with UMMB supplementation suggested that creating low nutrient contents in the diet through reducing concentrate feed and good quality roughage could be fulfilled through provision of UMMB and night feeding of rice straw. References Kunju, P.J.G. 1986. Urea molasses block lick: a feed supplement for ruminants. 261 274, in: M.N.M. Ibrahim & J.B. Schiere (eds). Rice straw and related feeds in ruminant rations. Proceedings of an International Workshop, Kandy, Sri Lanka, 2428 March 1986. Leng, R.A., 1983. The potential of solidified molasses based blocks for the correction of multinutritional deficiencies in buffaloes and other ruminants fed low quality agro industrial by-products. In: The Use Nuclear Techniques to Improve Domestic Buffalo Production in Asia. IAEA, Vienna. 135-150. Saddul, D. and A.A. Boodoo 2001. Response to urea molasses multinutrient blocks as a supplement in the diet of goats. Agricultural Research and Extension. www.gov.mu/portal/sites/ncb/moa/farc/amas2001/pdf/p1.pdf Sansoucy, R. 1995. New developments in the manufacture and utilization of multinutrient blocks.. FAO Animal Production and Health paper No.82. Rome, FAO. 78-83 Weerasinghe, W.M.P.B., S.S.P. Silva, A.C.M. Faizal, M.W.C.D. Palliyeguru and N. Priyankarage 2004. Formulation of cement free urea molasses multinutrient block for ruminants. Sri Lanka Veterinary Journal 51:(1)28. Weerasinghe, W.M.P.B., S.S.P. Silva, N. Priyankarage, U.L.P. Mangalika and R.A.T. Chandima 2010. Effects of supplementation of nitrogen through urea molasses multinutrient block (UMMB) on the perfiormance of dairy cows fed with good quality forage based diets. Abstracts of the 5th International Nitrogen Conference, 3-7 December 2010, New Delhi, India. 419. Wongnen, N. 2007. Feed supplementation of dairy cattle with UMMB in the northeastern region of Thailand. In: Feed supplementation blocks. Urea-molasses multinutrient blocks: simple and effective feed supplement technology for ruminant agriculture; FAO Animal Production and Health Paper (FAO). 111-124. www.vet.chula.ac.th/~nuclear/symposium44/Narong.htm Acknowledgements We would like to express our warmer thanks to all the staff members and laboratory assistants of the chemistry laboratory of the Veterinary Research Institute, Gannoruwa, Peradeniya, for providing us with expertise guidance, valuable information and necessary facilities to carry-out the research.
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Evaluation of Salmonella Cross Contamination at Retail Chicken Meat Outlets in Kandy Area
U.S. Alwis , D.C. Mudannayake, D.K.D.D. Jayasena Uva Wellassa University, Badulla, Sri Lanka and J.K.H. Ubeyarathna Central Veterinary Investigation Centre, Veterinary Research Institute, Gannoruwa, Sri Lanka Introduction Salmonellosis is among the most frequently reported food borne disease worldwide. While numerous potential vehicles of transmission exist, commercial chicken meat has been identified as one of the most important food vehicle for Salmonella (Abd El-Malek et al, 2011). The risk in different countries varies according to the control measures and the practices implemented along the food chain from primary production to final preparation of the meat for consumption (FAO/WHO, 2010). The prevalence of Salmonella contamination in poultry meat obtained at retail grocery stores is a better indicator of the public health risk. Transportation, handling, storage, additional processing and re-packaging of raw poultry meat products often occur, after the products leave the processing plant or after the animal is slaughtered at retail shop itself. Each of these steps may provide a new opportunity for bacterial contamination or growth. Salmonella in retail chicken meat outlets could be attributed to lack of proper cold chains, inadequate power supply and low level of hygiene in retail outlets. Therefore, the current study aimed at evaluating the Salmonella cross contamination at retail chicken meat outlets in Kandy area. Furthermore this study focused on making recommendations to improve quality assurance to mitigate the risk to consumers by identifying the risk factors associated with Salmonella cross contamination in retail chicken meat outlets. Methodology Samples were collected during the month of April 2011. Swabbing process Sterilized swabs were lightly moistened in saline water swabbed along the area of contact surfaces, utensils to recover the microbes. Swabs were placed in labeled containers and returned to the laboratory in an iced (at 4 C) Styrofoam box. All swab samples were processed immediately after reaching the laboratory. Data collection Fifteen (15) retail shops were selected to collect data based on their type of sales, sales scale, type of products, nature of sales, type of meat display, storage conditions of meat, way of handling meat, source of hygienic condition and etc by using pre prepared questionnaires. Salmonella Isolation Protocol Each swab sample was pre-enriched at 37 C in 5 ml of buffered peptone water (BPW) and placed in incubator overnight (ISO 6579). 0.1ml of each sample of this stock was transferred to 10 ml of Rappaport Vassiliadis (RV) medium. The RV media were then
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incubated at 42C for 24 hours. One loopful of RV media from each sample was streaked onto Xylose Lysine Deoxycolate (XLD) agar plates and the plates were incubated at 37 C for 24 hours. Presumptively positive black colonies were subcultured on Brilliant Green (BG) agar plates and incubated at 37 C for another 24 hours. Presumptively positive pink colonies were bio-chemically confirmed with Triple Sugar Iron (TSI) agar, Simmons Citrate agar, Urease and SIM medium. Results Prevalence of Salmonella cross contamination at retail chicken meat outlets in Kandy area Out of 57 swab samples collected, 12 swab samples of Salmonella suspected isolates from selective media were bio-chemically identified as Salmonella. Therefore the overall prevalence of Salmonella in retail chicken meat outlets was 21% (95% C.I 11.37- 33.88). Weighing scale (33%), Meat containing trays/buckets (27%) and Cutting board (25%), showed the highest percentage of Salmonella prevalence. Knife (14%) and showcase (9%) showed relatively low percentage of Salmonella prevalence. Prevalence of Salmonella cross contamination at retail chicken meat outlets with slaughtering facilities versus without slaughtering facilities Two types of retail chicken meat outlets were observed during the study. Retail outlets without slaughtering facilities represented 60% of the sample and retail outlets with slaughtering facilities represented 40% of the sample. Retail chicken outlets with slaughtering facilities had a significantly higher prevalence of Salmonella positive samples than retail chicken outlets without slaughtering facilities (p <0.05).The retail outlets which had no cooling facilities for raw chicken meat during display, had a significantly higher risk of Salmonella prevalence than the other type of retail chicken meat outlets with cooling facilities during displaying meat (p <0.05). Meat stored at room temperature during display was 13 times more likely to be contaminated with Salmonella than meat stored at cooling temperature during display (C.I. 0.88-207.63). Table 1: Multivariate regression analysis of frequency of cleaning, use of detergents and use of disinfectant as risk factors for Salmonella contamination at retail chicken meat outlets Predictor Frequency of cleaning Use of detergents Categories Two times per day Less than two times Yes No Use of disinfectant Yes No Statistically significant at ( =0.05) No of outlets 7 8 9 6 5 10 % 46.7 0.46 53.3 60 40 33.3 66.6 0.062 P
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Risk analysis for Salmonella cross contamination at retail chicken meat outlets Factors that were included in the risk analysis were not significantly associated (p > 0.05) with the Salmonella cross contamination. Discussion The present study demonstrated an overall Salmonella cross contamination of 21% for the retail chicken meat outlets in Kandy area. This level of contamination is lower than with the recent literature from countries like Vietnam 53.3% (Van et al., 2007), India 65.71% for raw chicken breast and 71.43 for raw chicken thigh (Wilfred and Nadeem, 2011), Canada 30% for raw chicken legs (Bohaychuk et al., 2006), but similar to Maryand 23% for raw chicken meat (Myint, 2004). The differences in the prevalence of Salmonella in raw poultry may be due to country of origin, type and size of sample analyzed, and methodology (Bohaychuk et al., 2006). The differences in the media used for enrichment, selective enrichment, and isolation can also affect estimates of prevalence (Myint, 2004). The different rate of contamination in different countries can also be related to climate and temperature of storage of raw meat at retail meat outlets. Better equipment in slaughterhouses, advanced processing practices (including the use of dry chilling of carcasses), and more effective use of refrigeration in meat transport in developed countries could also help to reduce cross contamination of meat (Van et al, 2007). When high moisture niches are present Salmonella may be a significant hazard. The survival and the growth of microbes in a food processing environment may lead to contamination of the finished product. Cleaning and disinfecting procedures for processing, handling, and holding equipment may be critical control points for prevention of post processing recontamination. The requirements for meat handling at retail, it is recommended that hygiene measures should be aimed at minimizing cross contamination between raw chicken and hands, contact surfaces and utensils. The true incidence of salmonellosis is difficult to evaluate because of lack of an epidemiological surveillance system in the country. The absence of centralized slaughter facility and the small volume of retail business, unstable policy, rules and regulations and the knowledge gap are the hurdles for hygienic production of chicken meat at retail outlets in the country. The conditions during slaughtering, processing, transportation, holding at retail outlet, and opportunities for cross-contamination at retail outlets can result large changes in the risk of salmonellosis to the consuming public. A greater potential risk is that slaughtering animals at same place where all evisceration and cleaning take place, is used for handling, packing, and even further portioning of the final product at the retail grocery store outlet prior to sale to the consumer. Conclusions This higher rate of Salmonella cross contamination at retail chicken meat outlets could be attributed to lack of proper cold chains, and minimal facilities and poor level of hygiene in retail outlets. The findings of this study are vital to the public health risk of the country. Therefore, Sri Lanka needs its own unique model of development to assure the quality and safety of poultry products at retail outlets.
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References Bohaychuk, V. M., G. E. Gensler, R. K.King, K.I. Manninen, L. M. Mcmullen, O.Sorensen, M. E. Stiles and J. T. Wu 2006. Occurrence of Pathogens in Raw and Ready-to-Eat Meat and Poultry Products Collected from the Retail Marketplace in Edmonton, Alberta, Canada, Journal of Food Protection, Vol. 69(9):21762182. Coloe, P.J., T.Istivan, G. Moutafis and T. T. H. Van 2007. Detection of Salmonella spp. in Retail Raw Food Samples from Vietnam and Characterization of Their Antibiotic Resistance, Applied and Environmental Microbiology, 73:68856890. FAO/WHO. 2010. Proposed draft guidelines for control of Campylobacter and Salmonella spp in chicken meat, Report of joint FAO/WHO Food standards programme, Codex committee on food hygiene, 42 Session, Kampala, Uganda, 29 November 3 December 2010. Hassanein, R., S. F. Hassan Ali 2, A. M. M. A. Abd El-Malek Mohamed and K. I. Elsayh 2011. Detection and identification of Salmonella species in minced beef and chicken meats by using Multiplex PCR in Assiut city, Veterinary World, Vol.4 (1):5-11. Myint M. S., 2004. Epidemiology of Salmonella contamination of poultry meat products: knowledge gaps in the farm to store products, Ph.D. Dissertation: University of Maryland, Maryland, USA. 86-98. Ruban, S. W. and N. Fairoze 2011. Effect of processing condition on microbiological quality of market poultry meats in Banglore, India, Journal of Animal and Veterinary Advances, 10(2):188-191.
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Establishment of Community Based Fish Factory Through Green Supply Chain Management Approaches
A.D.Wijenayake Uva Wellassa University, Badulla, Sri Lanka Deepa Gamage Industrial Development Board, Galle Road, Katubedda and S.C.Jayamanne Uva Wellassa University, Badulla, Sri Lanka Introduction Post harvest loss is one of the main problems in Sri Lankan fish industry. According to Ministry of Fisheries and Aquatic Resources there is a 30% of post harvest loss in Sri Lankan marine fish industry. This may be due to lack of facilities and lack of knowledge of the fishermen. Under the greening concept the main idea is to increase the resource utilization by maximizing the output and reduce the environment impact. Therefore, by applying the greening concept the post harvest losses can be reduced and the environment effect could be minimized and maximum gain could be obtained from the existing resources. Establishment of a community based fish processing factory through green supply chain management approaches is tested here as an option to minimize the post harvest losses in Sri Lankan fish industry. Methodology As secondary data; type and quantity of fish production in each district, import quantity of the fishery products were collected from the data base of Ministry of Fisheries and Aquatic Resources (MoFAR), Ceylon Fisheries and Harbour Cooperation (CFHC) and customs reports. As primary data; supply chain of the fish, Fishing gear, storing condition on boat, average experience in fishing, temperature after unloading, time period of fishing and total fish catch were collected by using structural interview method. The raw material flow and the material balance of the fish canning factory also were identified to get an idea about the type and the amount of waste and the environmental impact of those wastes. Then possible solutions were to minimize those effects under the green supply chain management approaches was established. After identification of supply chain; regression model was designed to identify the factors affecting the fish quality. Fish quality was considered as the dependent variable because if the fish quality is high there may be fewer post harvest losses. Storing condition on boat, average experience in fishing, temperature after unloading, time period of fishing and total fish catch were considered as independent variables. Negombo, Beruwala, Dondra, Trincomalee and Kalpitiya harbours were selected by stratified sampling method, having highest number of multi-day boats. Those harbours were visited to collect fifty samples (ten samples from each harbour), as primary data and to get detail information on fishing methods and post harvest losses. Then a proper site was selected and type of the fish was selected and assumed that the production will be 1500 cans per day. All other calculations were done according to this 144
assumption and the feasibility study was carried out finally to find out whether the project is feasible to be implemented. Results and discussion The first two stakeholders (fishermen and commission agent) were analyzed by developing a regression model.
Correlation Coefficientsa Unstandardized Coefficients Model 1 (Constant) Storing B 100.003 -2.531 Std. Error 8.011 1.415 1.248 .706 .489 .202 .002 -.113 -.087 -.386 .203 -.349 -.135 Standardized Coefficients Beta t 12.483 -2.211 -2.029 -4.798 3.131 -4.670 -1.804 .000 .032 .049 .000 .003 .000 .078 P
Fishing gear -3.128 Temperature -3.389 Experience 1.530 Time Period -.946 Total fish -.004 Dependent Variable: fish quality
According to the result fishing gear, storing condition in the boat, average experience in fishing, temperature after unloading, time period of fishing, are the factors which are significantly affect the fish quality (P<0.05). According to the collected data, identified what will the best under greening concept. If the fish quality is higher than 75% considered that those fish are very good and can be use in processing. From collected data found out the average of the Total fish catch, Time period of fishing and average fishing experience and find out the highest frequency of Fishing gear and storing method in the boat and when considering temperature after unloading, it is obviously to have less than 40C, if not histamine formation will be high. According to collected data identified that average fishing experience is about seven years. When experience is high they know how to catch the fish by avoiding damages and also how to store the fish with minimum damages. When consider about the time period of fishing, identified that 13 days are the best time period of fishing. So there will not be excess fish and there may be fewer damages to fish caught early. The best quantity of fish is estimated as 2700 kg. Pole and line method found to be the best fishing method. When considering fish storing method it is better to have racks so that there will not be excess fish and there may be fewer damages to early caught fish. Then fish processors were evaluated by identifying and quantifying raw material flow to gain the knowledge on the amount of inputs and outputs from each step. Environment impacts from each step were found out and possible solutions were suggested.
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According to the collected data Matara was selected as the best location for the factory having the highest fish production in Sri Lanka. Skip jack tuna (Balaya) was selected as the best species for processing. Canning industry was selected as the processing method by looking at the quantity of fish products imported. Proper factory layout was designed to minimize the waste and all necessary steps were taken to minimize the environment effect and finally to have safe and quality products to consumers. Finally feasibility of the project was studied under the greening concept. The Return On Investment (ROI) of the project is 83.42%. It indicates that the project is highly feasible to be implemented. Conclusions Establishment of a community based fish canning factory under green supply chain management approaches is a highly feasible project. According to the profitability statement the return on investment was 83.42% .indicating that the project is highly feasible. However, it is not possible to achieve 100% performance at the start. Therefore, it is proposed to achieve 60% performance in the first year, 80% in the second year and finally 100% performance in the third year. So the feasibility of the project is in first year is 50.05% and the second year is 66.73% and in the third year 83.42%. The figures indicate that the project is highly feasible. Fish quality is taken as the main factor which is affecting to the greening of supply chain. That means if the quality of the fish is high there will be less waste generation and less environment impact and also can use the existing resources efficiently. So the factors; Fishing gear, Storing condition in the boat, Average experience in fishing, Temperature after unloading, Time period of fishing and Total fish catch were identified to be affecting the fish quality. References Ceylon Fisheries Corporation (CFC) [Online] Available at: http://www.fisheriescorporation.gov.lk/ (accessed on 08.05.2011) Department of Fisheries and Aquatic Resources (DFAR) [Online] Available at:
http://www.fisheriesdept.gov.lk/
Geoffrey, R., Ames, 2002, Traditional and modern post-harvest technologies for increased food and supply from inland fisheries in Africa, Natural Resources Institute Chatham, UK. Mohanty, R.P., Deshmukh, S.G., 2008. Essentials of Supply Chain Management. 274280.
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Cost Reduction of Brine Shrimp by Replacing of Low Cost Live Cultures (Moina, microworms) for Fresh Water Fish Guppy (Poecilia reticulata).
G.W.H.P.N. De Silva, S.C. Jayamanne Uva Wellassa University of Badulla, Sri Lanka and M. Hewavitharana Tropical Fish International( Pvt). Ltd. Introduction Ornamental fish farming is an expanding industry and its global export trade has grown steadily and today it is a multimillion dollar industry in many countries (Andrews, 1990). Sri Lanka contributes approximately 1% of the world's demand for ornamental fish. The demand for the fresh water fish is quite does not meet the demand because there are so many constraints related with the fresh water ornamental fish farming. The major constraint is the cost of feed especially during the stage of the post larva and fry. Artemia (brine shrimp) nauplii is the most common live food used in commercial larviculture of fresh water ornamental fish (Dahlgren and Phang. 1985; Kim et al., 1996) and the cost of 400 g of cysts is nearly Rs.4000.00. The present study aimed to find a suitable low cost live food which can replace high cost Artemia in aquariums giving more profits to the ornamental fish traders. Two live food species, Moina and Micro worms, which can be reared easily with very low cost are selected for the study and their suitability in rearing post larval stage and fry stage of guppy (Poecilia reticulata) was tested under aquarium conditions. Methodology The experiment was carried out at the Tropical Fish International Private Limited, Wagawatte, Horana. Mass culture of Moina and micro worms were carried out prior to conducting feeding trials. Three types of live food cultures namely, Artemia, Moina and Microworms were maintained for 21 days during feeding trials. Fifteen aquarium tanks of same size were used for the experiment and three tanks each were used for control and four treatments. Two hundred and fifty numbers of day-old fry were stocked in each tank and were fed six times a day according to the feeding schedule given in Table 1. Three types of live feed, Brine shrimp, Microworm and Moina and powder feed were used for feeding and the fry were fed ad libitum at each time. Table 1 Feeding schedule of the fish for control and 4 treatment tanks Control Treatment 1 Treatment 2 Time Feed Time Feed Time Feed 8.00 PF 8.00 M 8.00 M 9.00 PF 9.00 MW 9.00 PF 11.00 PF 11.00 M 11.00 MW 1.00 PF 1.00 MW 1.00 PF 3.00 BS 3.00 M 3.00 M 4.30 PF 4.30 MW 4.30 MW PF - Powder Feed BS - Brine Shrimp 147 Treatment 3 Time Feed 8.00 M 9.00 PF 11.00 MW 1.00 M 3.00 BS 4.30 MW M- Moina Treatment 4 Time Feed 8.00 M 9.00 BS 11.00 PF 1.00 M 3.00 PF 4.30 MW MW - Microworms
Water quality was checked daily at 7.30 a.m. before feeding the fish. He appearance of the water in the tanks was observed visually and the quality of water was ranked. Unionized ammonia, NH4+, pH, Nitrate (NO3-) was measured using test kits. Mortality was checked daily and recorded promptly. Lengths (mm) and weights (mg) were measured using a top loading balance (Sartorius) with a precision of 0.0001 and a Vernier caliper respectively in 100 fry randomly selected from each tank at weekly intervals. The differences in weight gain, mean weight gain, length, mean length, specific growth rate, condition factor, survival rate and water quality parameters ( water appearance, Ammonia, pH, Nitrate) were tested using one way ANOVA. Comparison of means was done by using the Turkey test to find out the significance between the means. The parameters were calculated according to the equations given below. W2 W1 F W2 - Weight of 100 fries+ water bowl, W1- Weight of bowl +water, F Number of fry Mean Weight = (W2 W1) W1
Mean weight gain = W2 -Final mean weight, W1-Initial mean weight Mean length =
L1 + + Lx F L Length of individual fish, F Number of fish fry Speci ic growth rate (SGR %) =( 2 1 100)/( 2 1)
Loge - log to base e, T2- time of final weight in days, T1- time of initial weight in days Survival rate (SR %) = Number of fry that survived 100 Number of fry stocked in the tank
Cost of feed for 21 day rearing period was calculated based on the cost of production for Moina and microworm and based on the market price of Artemia (Brine shrimp). The amounts used for daily feeding was recorded and the cost for each treatment was calculated. Results and discussion The growth performance of the guppy in control tank and four treatment tanks is shown in Table 2.
148
Table 2: The performance of guppy in control and treatment tanks (The values given are means of three replicates mean of three replicates Parameter Mean wt gain (mg) Final length (mm) Specific growth rate No.of fish stocked No. of harvested Survival rate (%)
Mean weight gain (mg)
Cont. 17.66+ 1.15 12.70 + 0.26 13.93 + 0.29 250 209.67+ 5.51 83.60 T1 19.00+ 2.00 13.63 + 0.23 14.25 + 0.47 250 222.33+ 3.51 88.8
Treatment Tanks T2 T3 16.33 + 22.66 + 1.15 0.57 10.40+ 15.53+ 1.15 0.25 13.577+ 15.202+ 0.32 0.23 250 250 185.67+ 7.23 74.0 236.00+ 3.00 94.4
T4 18.33+ 1.15 13.33+ 1.06 15.202+ 0.23 250 166.67+ 10.50 66.4
25 20 15 10 5 0 control trt.1 trt.2 trt.3 trt.4
Mean weight gain
Treatment tanks
Figure 2: Mean weight gain (mg) of guppy fish fry in relation to treatments
Specific growth rate%
15.5 15 14.5 14 13.5 13 12.5 Control Trt.1 Trt.2 Trt.3 Trt.4
Specific growth rate
Treatment tanks Figure 3: Specific growth rate of guppy fish fry in relation to treatments Treatment 3 showed the best mean weight gain, specific growth rate, length gain and survival rate (Figures 1 and 2 and Table 2) compared to other treatments. Therefore the combination of Moina, microworm, Brine shrimp is selected as the best feed among the tested feed for guppy fry. All treatments except treatment 2 showed a better growth and specific growth rate than control which is the currently practiced feeding strategy of the aquarium. Mean comparisons showed that there is no significant differenc between control and Treatment 1, 2 or 4 but there was a significant difference between control
149
and the Treatment 3 (P<0.05). Treatment 2 showed the worst results while other treatments fared well in comparison to Control. Reduction of feed cost was highest in Treatment 1 (95.88%) and 47.36% in Treatment 3. Cost for feed during the rearing period for control, T1, T3 were Rs. 820.00, Rs. 33.71, Rs. 431.18 respectively. Lowest cost was observed in Treatment 1 where only Moina and microworms were used as feed. The cost reduction was 98% by using live feed as in Treatment 1 and 48% using feed as in Treatment 3 (all four types of feed). Conclusions The present study showed clearly that the cost incurred by aquarium traders using Artemia as live food for fry can be reduced using other low cost live food. Moina and microworms can be used in aquariums for feeding fry stages successfully. Different combinations of Moina and microworms could be used in this regard and the aquarium traders can select the suitable combination considering the cost. However, It is also advantages to feed Artemia once a day to gain better growth. Findings of this study will be useful for the development of fresh water ornamental fish farming in Sri Lanka. References Andrews, C. 1990. The ornamental fish trade and fish conservation. Journal of Fish Biology, 37: 53-59. Dahlgren, G.V. and V.P.E. Phang. 1985. Food and feeding behavior of the guppy, Poecilia reticulata (Pisces: Poeciliidae). Can. J. Zool. 59:684-701. Kim J., K.C. Masses, R.W. Hardy 1996. Adult Artemia as food for first feeding coho salmon (Oncorhynchus Kisutch). Aquaculture. 144:217-226.
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Preliminary Study on Effect of Different Feed Combinations on Captive Breeding of Anemonefish Amphiprion Clarkii
P.R.A. Pathirana, S.C. Jayamanne , N.P.P.Liyanage . Uva Wellassa University. Badulla. Sri Lanka and J. P. R. P. Kumarasinghe Ceylon Aquatics (Pvt) Ltd., Galayaya, Pannala, Sri Lanka Introduction The marine ornamental fish trade began in the 1930s in Sri Lanka (Buckner, 2004). Harvesting marine species for home aquaria has started in 1980s (Andrews, 1990) and the exports have continued to increase in 1990s (Vallejo, 1990). The trade has expanded to a multi-million dollar business and 45 countries supply global markets an estimated 14-30 million fish annually. The largest suppliers are Indonesia and the Philippines, followed by Brazil, Maldives, Vietnam, Sri Lanka and Hawaii. Approximately 150 species of marine fishes are exported from Sri Lanka and all these come from the wild catches. Even though Sri Lanka has a vast potential for marine ornamental fish trade, it has not developed technology on breeding marine ornamental fish in captivity. Anemone fish, Amphiprion clarkii is a species which has a high demand among marine aquarists due to its attractive colours and behavioural display. The fish is caught from the wild destroying the natural habitats due to improper catching methods and may decrease the population. The genus Amphiprion represent the most important group of captive bred marine species (Olivia et.al, 2006) and the present study aimed to find the possibility of stimulating breeding in Amphiprion clarkii in captivity using two different feeds to reduce the pressure on the natural environment. Methodology Four glass tanks of the size (91.5 cm X 47 cm X 38 cm) were used for the study and all the tanks were set up in a same height providing equal amount of light and temperature. The bottom of each tank was filled with same amount of cleaned coral sand and gravel just enough to cover the bottom. Two cleaned clay pots were placed in each tank providing hiding places and a substrate to deposit their eggs. Each tank was connected to a triple pass type protein skimmer (400 l per hour) and a biological filter (Figure 1). All the tanks were supplied with aeration and were numbered. Purified and disinfected sea water was transported to Pannala from Marawila area. Each tank was filled with a volume of 129 l sea water and recirculation system was in operation throughout the study. Salinity of the water was adjusted around 30 31 ppt. Four pairs of anemone fish (Male: around 4 cm, Female around 8 cm) paired out naturally were obtained from the coral reef environment of Tricomalee sea. One pair of fish was introduced to each tank after circulating the water system for 24 hours. Two different feeds were prepared to feed the fish as formulated feed and the mussels. Mussel feed was prepared by grinding cleaned mussels and the formulated feed was prepared by grinding the cleaned ingredients; fish (50%), seaweeds and prawns (20%), cuttlefish (15%), mussel meat (15%) with garlic. Feed preparation was done bi-weekly. Tank number one and three were fed with a formulated frozen feed and tank number two and four were fed with mussel meat at ad
151
libitum for three times per day. Changing and siphoning of water was done once a week and salinity was adjusted around 30 ppt. Fish were observed three times per day for their behaviors and all the information was recorded. Tanks were specially checked for eggs in the morning and afternoon. Salinity and temperature in the tanks was checked three times a day using a digital salinity meter and a temperature meter (YSI 30) respectively. Ammonia levels were checked with an ammonia meter (HANNA: HI 96733) twice/month and the level of ammonia was maintained between 0 - 0.001 ppt. Data were collected for three months and were analyzed with two proportion z test in MINITAB 14 statistical package.
Figure 1: The water filtering and circulating system Results The fish that were fed with formulated feed diet started pre-spawning behavior after eight weeks after stocking while those fed with mussel did not show any spawning behavior. During the pre-spawning period fish rarely swam around the tank and at first, the female selected a specific place and stayed there and after several days same place was occupied by the male. Then, cleaning of the substratum was started by the female and later both male and female engaged in cleaning. The results indicated that formulated feed has a significant effect (P<0.05) on stimulating spawning behavior in A. clarkii. The pre spawning behavior was limited to a period of fourteen days but spawning did not materialize. The environmental factors, salinity, ammonia, nitrate and nitrite remained constant during the experimental period but the temperature has shown fluctuations. The temperature showed a significant effect on the pre-spawning behavior (P<0.05) and the pre-spawning behavior was interrupted when the temperature increased greater than 27 0C. Discussion The results of the study has shown that the formulated feed has a significant effect (P<0.05) on the breeding of Amphiprion clarkii. When the temperature level began to increase the pre spawning activities were stopped by the fish. According to the previous records, Dalia and Svedang (1997) has shown that water temperature has a very marked effect on the psychological and biochemical process in fish, and a raised temperature regime has a complex effect on fish reproductive, nerve and endocrine system. The
152
temperature presumably effect on both GtH (Growth Hormone) secretion and the responsiveness of target organ to hormonal stimulation. Increased temperature affects the fat synthesis, metabolism and endocrine system which results in the failure of the generative processes. The histological analysis revealed high frequencies of egg resorption and the gonads developed arhythmically (Dalia and Svedang, 1997) . Most of fish including Amphiprion clarkii, are external fertilizers. The external environment should have the ability to protect the eggs with suitable conditions. When the environmental factors are suitable, gametes are released to the environment by fish after having several behaviors (Pre spawning behaviors). These behaviors are induced by the endocrine state of the fish. According to Dalia and Svedang (1997) if the environmental temperature is raised, it is affected negatively to the endocrine system of the fish. According to Wood and McDonald (1996) there is a close association between reproductive behaviors and endocrine state, and any environmental factor (i.e. Temperature) that interferes with normal endocrine functions may also disrupt behavioral processes. Conclusions The formulated feed used in this study is a good source of nutrients as a brooder feed in breeding anemonefish, Amphiprion clarkii in captivity. The water circulating system which was used in tanks kept low levels of dissolved compounds and ammonia is efficiently removed by the system. The fish can survive without any disturbances up to 300C, but the spawning activities has not taken place in temperatures above 270C and higher temperatures affected spawning activities of Amphiprion clarkii negatively. The results are encouraging and need further research to succeed in breeding of Amphiprion clarkii in captivity.
References Andrews, C., 1990. The ornamental fish trade and fish conservation. J. Fish. Biol. 37: 53-59 Bruckner A.W., 2004. The importance of the marine ornamental reef fish trade in the wider Caribbean. NOAA Fisheries, Office of Habitat Conservation, 1315 East West Highway, Silver Spring, MD 20910, USA Dalia, L. D. and Svedang, H. 1997. A Review on Fish Reproduction With Special Reference to Temperature Anomalies. 10:14-17 Olivia, J., Fernando K., Raja and Balasubramanian T. 2006. Studies on Spawning in Clown Fish Amphiprion sebae With Various Feed Combination Under Recirclating Aquarium Conditions. 376-381. Vallejo, V. B., 1997. Survey and revive of the Philippine marine aquarium fish industry. 10: 25-26. Wood, E. 1985. Exploitation of Coral Reef Fishs from the Aquarium Trade. 121 Wood, M. C. and McDonald G. 1996. Temperature Effects on the Reproductive Performances of Fish. 159-176.
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Production of Tuna Fish Oil by Utilizing Tuna (Thunnus Albacares) Processing by-Products
M.S.S. Kumara Uva Wellassa University, Badulla, Sri Lanka G. Rajapakshe Jay Sea Foods (Pvt.) Ltd, Ja-Ela, Sri Lanka and S.C. Jayamanne Uva Wellassa University, Badulla, Sri Lanka Introduction Sri Lanka is surrounded by a coastline of approximately 1700 Km, and belongs to an Exclusive Economic Zone (EEZ) of 517,000 Sq Km. About 50,000 people in the country are directly involved in the fishery industry. The total marine fish production in 2010 was recorded about 332,260 Metric tons (Mt), while Tuna contributed 88903 Mt to the total (Fisheries year Book, 2010). From the total yield about one-third of the catch of fish is not used for direct human consumption but for the production of fishery by products (Balios, 2003). Every year thousands of tons of fish by-products of high nutrient content are discarded by fish processing plants through the world although they can be utilized for other purposes. Crude tuna oil is produced from tuna waste by steam followed by purification, wet rendering, alkali digestion, acid silage Soxhlet like methods (Bimbo, 1990).Tuna fish oil has been considered as an available source of long chain polyunsaturated Omega 3 and Omega 6 fatty acids, especially eicosapentaenoic acid (EPA) and Docosa Hexaenoic Acid (DHA). Tuna oil differs from other fish oils in the ratio of the C20:5 n-3 (EPA) to the C22:6 n-3 (DHA) fatty acids. It means that the ratio of EPA: DHA in tuna oil around 1:4 is similar to that of human breast milk . This study attempted to find out the feasibility of producing Tuna fish oil using fish waste. Methodology Heads of yellow fin tuna (Thunnus albacares) weighing more than 20 kg were obtained from tuna processing plant in Ja-ela, Sri Lanka Selected tuna heads were separately crushed up using a mechanical crusher to get fine ground particles. Tuna fish oil was separated using Soxhlet method, wet rendering method and alkali digestion method respectively. In Soxhlet method 100 g of sample of prepared tuna head sample was transferred into extracting thimbles. Oil was extracted with petroleum ether (chloroform and methanol at the ratio of 2:1). The temperature was maintained between 60 OC to 80 OC for 6 hours and extracted fish oil was weighed. Wet rendering method was carried out according to the Bimbo (1990). A sample of 100 g of prepared tuna head were mixed with 20 % water and heated by using a water bath maintaining the temperatures 75 OC, 85 OC and 95 OC each with heating times of 10 minutes, 20 minutes and 30 minutes followed by pressing. In Alkali digestion method, crushed tuna head samples were taken (100 g) and mixed with 10 % of distilled water and digested for 30 minutes at 50 60 OC. After 30 minutes 20 mL of 2% Sodium hydroxide was added slowly to avoid soap formation. The mixture was then cooked at a temperature of 80 OC up to 30 minutes with constant 154
stirring until the head muscle particles were turned semi colloidal state. After 30 minutes oil was separated using centrifugation. Free fatty acid analysis was made by extracting free fatty acids in hot alcohol and titrating with standard alkali medium. Results Results of the analysis of oil, using different extraction techniques, are presented in Table 1. Table 3 Yield of oil extracted using different extraction methods Parameter Yield Free FA Value Soxhlet method 13.655 +/-0.113 .621 +/- .061 Wet Rendering 3.046 +/-0.509 .125 +/- .074 Alkali Digestion 5.296 +/-0.611 .0008 +/- .0012
Soxhlet method recorded the highest oil yield of 13.65 +/-0.113. followed by Alkali digestion method (5.29 +/-0.611). Wet rendering method showed the lowest oil yield (3.04 +/-0.509). All the mean values were subjected to One-way ANOVA to find the significant of the methods. A mean separation was done by using tukey test to find whether there is significant difference among the methods. Free fatty acid content of tuna fish oils separated by different methods is presented in Table 1. The lowest free fatty acid value was recorded from oil recovered by the alkali digestion method (0.004%). Wet rendering method was the next oil extraction method which reported the low free fatty acid value (0.125%). Soxhlet method recorded the highest free fatty acid value (0.62%).
0.7
Free fatty acid value
Free fatty acid values of different extraction methods 0.62
0.6 0.5 0.4 0.3 0.2 0.1 0 1 2 Extraction methods 3 0.125 0.0049
Figure 5 Free fatty acid value of soxhlet(1),wet rendering(2) and alkali digestion method(3).
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Discussion In this study tuna fish oil was extracted by using different methods and quality was determined by finding the free fatty acid values of the oil. According to the results of this study highest oil yield was obtained using Soxhlet method. Lipids will dissolve in a variety of solvents. Solvents such as chloroform and methanol with somewhat higher polarity have high dissolving property and Soxhlet method has reported the highest oil yield. In alkali digestion method fish tissues are digested with the help of alkali medium (sodium hydroxide 2%) and liberate oil from the fish tissues easily. Application of heat accelerate the digestion process to some extent, but release of complex lipids to the solution is lower than that of the Soxhlet method (Tanikwa, 1971). Wet rendering method was carried out according to the method of Bimbo (1990). The highest oil yield was observed at 85 OC for 20 minute treatment and lowest oil yield observed in lower temperature treatment (75 OC) and also high temperature treatment for long time (95 OC for 20 min, 30 min) method. This is possible due to the fact that lipid cells were ruptured to a great extent with 85 OC (Bimbo (1990). Among the different time temperature combinations the optimum condition for tuna oil extraction was observed to be heating the sample at 85OC for 20 min. The highest free fatty acid percentage in oil was recorded from Soxhlet method and alkali digestion method recorded the lowest free fatty acid value. According to the oil prepared at higher temperatures had higher free fatty acid amount. This results indicated that the hydrolysis of ester bonds of triglyceride occurred less at lower temperatures as well as oil can undergo hydrolysis in the presence of moisture and heat. In Soxhlet method oil is kept at somewhat higher temperature for long time (50 -60 OC for 6 hours) and hydrolysis of triglyceride was high resulting high free fatty acid value. Second highest free fatty acid value was recorded in wet rendering method, because a high temperature (85 OC for 20 min) was used. Alkali digestion method gave the lowest free fatty acid value. In alkali digestion method Sodium hydroxide add to digest the fish tissues so it will neutralize some amount of free fatty acid in the oil. Hence the Soxhlet method is recommended for extracting oil from fish. References Bimbo, A.P. 1990 . Production of fish oil in M.E. stansby,fsh oil in nutrition (pp.141180). New york:Reinholdpublishing Co.ltd
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Proceedings of the Research Symposium of Uva Wellassa University, December 15-16, 2011

Development of an Egg Based Sausage

Development of Ready to Eat Marinated Chicken Parts

Microbiological Quality Assessment of Raw Milk to Identify Sources of Contamination

vegetative pathogens and spores outgrowth

Microbial Cross contamination

Development of Chocolate- Malted Whey Beverage Using Liquid Cheese Whey

40 35 30 25 20 15 10 5 0 0 1 4 6 8 11 Storage period (days) Ginger incoporated milk Control

5% sugar 10% sugar

7.5% sugar Control

Figure 3. Variation in the sensory attributes of milk samples

Figure 4. variation of SPC in ginger flavored milk and control

Development of Egg Less Cake Incorporating Yoghurt

Locals Jamnapari Cross Total frequency of each pattern

Natural breeding AI program

Flock performance hatchability

30 25 20 15 10 5 0 1 2 3 weeks 4 5 Natural Breeding

Natural Breeding Aritificial Inseminati on

Body Weight Gain (g)

Feed Intake (g)

VAR00001 Dependent variable/Uniformity VAR00002 (Error) Stocking Density VAR00003 Temperature

0.45 0.24 0.24 0.43 0.93 0.42 0.51 0.310.34 2 3 4 5 6 7 8 9 Treatments

Feed Intake (g)

25 20 15 10 5 0 control trt.1 trt.2 trt.3 trt.4

Mean weight gain

15.5 15 14.5 14 13.5 13 12.5 Control Trt.1 Trt.2 Trt.3 Trt.4

Specific growth rate

Free fatty acid values of different extraction methods 0.62

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