September | October 2011 Feature title: Challenges associated with carrying out a meta-analysis of essential amino acid requirements

of fish
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F: Amino acid

Challenges associated with carrying out a meta-analysis of essential amino acid requirements of fish
by Guillaume Salze*, Margaret Quinton and Dominique P. Bureau, UG/OMNR Fish Nutrition Research Laboratory, Department of Animal and Poultry Science, University of Guelph, Guelph, ON, Canada, N1G 2W1 Email: gsalze@uoguelph.ca

igh cost of fishmeal, the volatility in the price of agricultural commodities and the stagnant price of aquaculture products are constraining aquafeed manufacturers to pay very close attention to the cost-effectiveness of their feeds.

fish and penaeid shrimp over the past 50 years and the body of knowledge on EAA requirements of aquaculture species is continuously expanding. One of the problems resides in keeping up with progress and developing a wholesome understanding of the state-of-the-art. The great diversity of methodological approaches used and animal species and ingredients studied as well as the multitude Nutritionists are required to formulate of opinions with regards to optimal levels to lower or narrower essential nutrient and modes of expression of EAA requirespecifications in order to minimise feed ments limits the ability of manufacturers to cost. At the same time, these feeds must meaningfully improve the cost-effectiveness sustain high growth, feed efficiency, health of feeds and/or adapt formulations to an and product quality of the animals at the ever changing commodities market. farm. A number of scientific reviews and Nutritionists also are relying on an publications have attempted to summarise increasingly diverse portfolio of economithe body of knowledge on EAA nutrition cal protein sources, with different amino and requirements of aquaculture species acid profiles. (Wilson, 1989; NRC, 1993; Cowey, 1994; Lall Thus, formulating cost-effective aquaand Anderson, 2005; Bureau and Encarnao, feeds requires increasingly precise infor2006; Hernandez-Llamas, 2009; NRC, 2011), mation on essential amino acid (EAA) and some concluded that the state-of-therequirements of aquaculture species. art on EAA nutrition of aquaculture species is still quite shallow. Dynamic field of research The choice of the mode of expression Aquaculture nutrition is a very dynamic (percent dry diet, percent crude protein, field of research. A very large g/kJ digestible energy, ideal protein, etc.) number of studof EAA requirement is a matter of much ies have been debate, and reflects the conflicting assumpconducted tions authors make when considering what on EAA affects the requirements or not (Bureau and nutriEncarnao, 2006; Bureau, 2008). tion of Together with methodological issues teleost (notably limitations of the Figure 1: Diagram illustrating the screening of studies, from experimental the total number found to the design used), number of suitable studies that the variability in constituted our working data set achieved growth

and feed efficiency, as well as differences in the mathematical and statistical approaches used to analyse data, these result in high variability in estimates of EAA requirements. Understanding the reasons underpinning this great inconsistency is important for developing more reliable and practical estimates of EAA requirement of aquaculture species. All these issues point toward a need for the systematic integration and analysis of information from the large number of studies that have been published so far on EAA requirements of aquaculture species. Statistical meta-analysis offers a mean to realize by integrating and standardizing information and allowing meaningful comparisons. The goal of this project was to carry out a meta-analysis of EAA requirement of fish through the construction of a dataset gathering all available data on EAA requirements of teleost fish.

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Figure 2: representation of EAA across the dataset

28 | InternatIOnal AquAFeed | September-October 2011

F: Amino acid

the accuracy of the non-linear regress i o n s analyses. Studies had to report information on w a t e r t e m perature , Figure 3: representation of species across the datase experiment duration, diet composition (dry-matter basis), initial The main objectives of this effort were and final individual body weight, and feed identifying factors that may affect estimates intake. Screening of the studies with of requirement, highlighting the shortcomthese selection criteria yielded a final ings in the existing body of knowledge, and dataset comprised 109 studies, which providing guidelines for future research. covered all 10 EAA in 28 teleost species (see Figure 1). Building a working dataset A comprehensive search of papers on EAA nutrition of commercially relevant A fragmented & diluted teleost species (for example, salmonidae, body of knowledge cychlidae, cyprinidae) published in peerIt is striking that less than half of the 249 reviewed journals and other technical puboriginal studies could be considered for the lications was carried out. working dataset. This search yielded 286 papers of which This highlights the limited scope of many 249 were original research studies focusing studies and/or scientific manuscripts and on EAA requirement of teleosts. the uneven quality of the research effort. As expected, a great variety of objecAn important cause of rejection of studies tives, experimental designs, and analytical was simply a lack of reported information, methodologies were employed in these which precluded us from calculating the studies. Selection criteria were therefore various variables. applied to the original dataset to identify Simple parameters, such as feed intake studies suitable for a meta-analysis. (feed served) and the dry matter content Amongst selection criteria, studies had of the diets, were frequently not reported to have at least five or more experimental by authors. Other major motives for diets with graded levels of an EAA. This rejecting studies included the use of too criterion was established in order to ensure few graded levels of nutrient studied or

EDITORS DESK

F: Aqua News Amino acid

be taken improve Aquaculture centre welcomes CFP toreforms the quality and Simple steps can ously highlighted he new Astec Aquaculture sented by Maria Damanaki, relevance of futurethe need for a in the sector, we ver y much Bureau, D.P., 2008. Toward a better definition coordinated approach to support welcome the EUs intention to Business & Science Centre Commissioner for Maritime studies and allow of essential amino acid requirements of fish, located on Nor th East Affair s and Fisheries, Astec us the develop moreof this dynamic remove administrative and legisto development International Aquafeed. Perendale Publishers Ltd, but fragmented industry, 14-17. so we lative barriers to trade. Englands Northumberland coast- welcomes the EUs recognition precise estimate pp. Mr Haddrick adds, Astec shares line, is a purpose-built facility, of the importance of developing of are par ticularly suppor tive of EAA requireCowey, C.B., the views requirements the Experiments which provides a unique and sup- ecologically viable aquaculture as ments.EUs plans to establish a new 1994. Amino acidof Commissioner of fish: a critical appraisal of that values. Aquaculture Advisor y Council Damanaki presentthere is great portive environment for all types should not ideally to give than and Aquaculture 124, 1-11. of aquaculture related business use fewer advice six guidance on potential within the aquaculture sector. industry-related issues.Lall, S.P., Anderson, S., 2005. Amino acid nutrition and research activities. experimental diets, We look forward also of salmonids: It has a specific focus on helping and The repor tsuf- states that Dietary requirements and to seeing report Figure enterprises arginine requirements for a information for aqua- how the framework develops fledgling 5: Estimatedto become ficientbetter framework bioavailability. Cahiers Options Mditerranennes rainbow trout grow, and the 63, 73-90. established andaccording to as mode of expression on culture composidiet will increase production and strongly suggest the EU grasp and supply seafood in the EU, this opportunity to ensure aquasuch, provides a unique combition, growthof perHernandez-Llamas, A., 2009. Conventional and reduce dependence onalternative dose-response models to estimate imported culture plays an integral part in formances and husbandry information. 14nation of plug-in for arginine requirement and 23 percent and go facilities fish and boost growth nutrient requirements of aquaculture species. in coastal developing a more sustainable The BLM has now been clearly demof and specialist business suppor t rainbow trout). approach and rural areas. services. onstrated to underestimates requireThis shows that much remains to be Aquaculture 292, 207-213. to fisheries manageBy 2014, Member States will ment. Astec reconcile the existing data and ments, and thus should be avoided. done to has a very unique offering NRC, 1993. Nutrient Requirements of Fish. that appeals to underpinning variability. Instead, the QM or draft national strategic plans to BQM offer a determine factors marine scientists National Acad. Press, Washington, DC. remove administrative barriers More inforMation: and industry alike a year round points better balance between accuracy and Nevertheless, several valuable and uphold ease supply of near tropical temperNRC, 2011. Astec practicality (that is, parsimony and environmental, social Nutrient requirements of fish arose from the data standardisation. and shrimp. National Academies and economic standards for the TheLyne Sands, Woodhorn Press, ature, flow-throughmodels was impactof fit). The accuracy of the seawater, farmed-fish industr adds the Northumberland Most importantly, studies should be y, Washington, D.C., 360 pp. edenablingshape of the curveaquatic meant by the a wide range of they are NE63 9NW part proposals to manage clear to plants and animals to be culti-based onof itsdesigned so results fit areport. diminishfit. Since all of these models are Wilson, R.P., 1989. Protein and amino acid United Kingdom fish stocks.ing return pattern. The As the dietary aquaculture ofTel: +44 1670 852771nutrition. In: vated there. graded UKs first requirements fishes. Progress in fish the law of diminishing returns, they attempt Kevin levels chief executive Further to the recent Common should include to fit curves shaped as those illustrated in Haddrick, of the test EAAbusiness incubator, with a focus Email: info@astecaquaculture.com Shiau, S.-Y. (Ed.), Proceedings of the fish nutrition businesses Website: Fisheries Policy Reform, pre- of Astec says, We have previ- on supporting start-up symposium, pp. 51-77. www.astecaquaculture.com clearly adequate levels so an obvious Figure 4. plateau can be Therefore experiments should be observed. designed to produce such result curves. COMPLETE PLANTS AND MACHINES FOR THE FISH FEED INDUSTRY Concomitantly, For example, data points of a given study equally clearly will approach the horizontal line if there is deficient levels little difference between treatments, which Good Flat Die Pelleting Presses Agricultural should also be will cause the model to fail. Practice n intern iels Otto Damholt g o u n for small, to Our meta-analysis shows that if therfinal d i included a t i o n a l

(a)

(b)

(95%)

poor growth performance achieved during the trial. The large number of species studied and the large number of EAA resulted in a very fragmented dataset (see Figures 2 and 3). For example, the dataset only includes four studies on phenylalanine requirements, which were all conducted on different species. Great differences in the body weight of fish used (<1g to more than 600g) introduce some challenges for standardisation of data. Almost half of the studies did not report information on carcass composition. Protein gain (retention), an important response variable for EAA requirements, could only be computed for a limited number of studies. Finally, only 16 percent of the studies in the working data set included some evaluation of protein digestibility. Consideration of digestibility would greatly strengthen and refine our understanding of EAA requirements in fish by taking account of some of the variability due to bioavailability of EAA in different ingredients. In order to standardise the data onto a common ground, two different modes of expression (percentage of EAA of interest in the dry diet, amount of EAA per MJ of digestible energy) were computed and two different growth response variables (weight gain per kg of metabolic body weight and thermal-unit growth coefficient). Therefore, for each study we obtained four pairs of variables, each of which was analysed using four mathematical models: the broken-line model (BLM), the quadratic model (QM), the broken-quadratic model (BQM) and the saturation kinetic model (SKM) (see Figure 4) in each study to allow estimation of EAA requirements.

Nutricin Acucola, Universidad Autnoma de Nuevo Len, Monterrey, Nuevo Len, Mexico, pp. 29-54.

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(c)
(95%)

(d)

(95%)

Figure 4: Diagrams illustrating the typical shape of the four models: (a) brokenline model (BLM), (b) quadratic model (QM), (c) broken-quadratic model (BQM), and (d) saturation kinetic model (SKM). Dashed line indicates the determination of the requirement for each model

Figure 5 presents the computed arginine requirement for rainbow trout using six studies for which models fitted correctly, depending on the three modes of expression. It illustrates that even after selecting suitable studies and standardising data, very large discrepancies in estimates between the studies remain. Variations around estimates based on ingested EAA remain high (coefficient of variation between 20 and 35 percent overall,

ensure sufficient body weightjoins Hamlet Protein sales management in the of the slowest-growing group in 70 position the animal d i f f e sector s is higher than the percent of of fastest nutrition r e n c eto the company. sales group, between treatgrowing director. then the probability of ments receiving obtaining a good fit decreases significantly. The strengthening of suboptimal levels. When using the SKM, this number is closer Hamlet Proteins commercial organisation to 50 percent. Additionally, if the curve only comes as a result of References increases but does not plateau, then the significant increase Bureau, D.P., models can only guess where the curves in the demand for Encarnao, P., plateaus or peaks. Since the require- H a m l e t Adequately s 2006. P r o t e i n ment is estimated by the EAA level products during 2010 defining the amino Niels the curve plateaus or peaks, such andacid requirements 2011, and to where Otto Damholt of fish: The case an estimation should not be considered support the continued example of lysine. Based reliable. in Horsens, Denmark, quest to deliver the compaIn: Cruz Surez, nys strategic goals in the high Niels Otto Damholt will be incorpoStudies which simultaneously protein market. responsible for Hamlet Proteins quality rated very deficient and obviously adequate soyaL.E., Ricque Marie, D., Tapia Salazar, M., sales and customer clear diminishing levels of EAA produced a ser vice Nieto Lpez, M.G., outside NAFTA and will be return pattern and consequently had More inforMation: vastly Villarreal Cavazos, repor ting to chief commercial improved chances of estimating the requireSren Bank D.A., Puello Cruz officer of Hamlet Protein, Sren Chief Commercial Officer ment accurately. y Armando Garcia Bank. Hamlet Protein tega, A.C. Or Niels Otto and Conclusions Damholt, aged Saturnvej 51 (Eds.), Advances 47, joins Hamlet Protein recommendations from DK - 8700 Horsens en Nutricin Denmark aIn this study regional directorimportant position as we highlighted Acucola VIII Tel: +45 75 631020 in Danisco Animal estimates variations in requirementNutrition,despite +45 75 VIII Simposium an Fax: 631025 Internacional de br inging 20 year our back- Website: attempt to standardise s ofworking dataset. www.hamletprotein.com

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