ISSN 0974-0775

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GREEN FARMING
(International Journal of Applied Agricultural & Horticultural Sciences)
(Abbreviation : Green Farming Int. J.)

Volume 6 Number 3 May-June 2015 Bimonthly

Previous issue :
CONTENTS Vol. 6, No. 2, pp. 222-442
Research Papers
Determining path of productivity in derived F1 involving diverse heterotic groups of cotton (G. hirsutum L. )
? H.G. KENCHARADDI, R.R. HANCHINAL and S.S. PATIL ......... 443
Genetic variability in Gossypium hirsutum cotton for seed cotton yield & fiber quality traits under organic cultivation
? I.S. KATAGERI, H.B. BABALAD, SURESH HANDI and R.F. CHANNAGOUDRA ......... 448
Heterosis & combining ability for grain yield and its component traits of newly developed inbred lines of
maize (Zea mays L.)
? K.L. NAVEEN KUMAR and G. SHANTA KUMAR ......... 452
Combining ability analysis for yield and yield contributing characters in pigeonpea (Cajanus cajan L. Mill sp.)
? A.H. GEDAM, A.B. BAGADE, A.B. JADHAV and I.A. MADRAP ......... 457
Variability, correlation and path analysis studies in sugarcane (Saccharum spp.)
? N.V. KHENI, S.C. MALI, M.M. PANDYA and Y.A. VIRADIYA ......... 460
Correlation, path and genetic divergence analysis in fodder bajra [Pennisetum glaucum (L.) R. BR.]
? HARINARAYAN BIND, D.N. VISHWAKARMA, BAUDH BHARTI, SANJEEV KUMAR, J.K. VERMA
and M.K. PANDEY ......... 464

Heterosis studies for yield and yield attributing characters in rice (Oryza sativa L.) for South Gujarat
? NAVIN CHANDER GAHTYARI, P.I. PATEL, DAYANAND and BALWAN ......... 468
Half diallel mating to study combining ability and heterosis for yield & yield contributing traits in bittergourd
? M. MAHESH, R.V.S.K. REDDY and P. SAIDAIAH ......... 474
Heterotic effects for seed yield & component characters in Indian mustard (Brassica juncea L.) C. & C.
? U.J. PATEL, K.V. PATEL, M.P. PATEL and J.A. PATEL ......... 478
Genetic variability, correlation and path analysis studies in safflower (Carthamus tinctorius L.)
? G.R. GOPAL, A.B. BAGADE, N.G. GAWANDE and S.B. DOIJAD ......... 483
Genetic diversity through metroglyph analysis & estimation of protein, iron and zinc in F5 progenies of a
wide cross of mungbean x blackgram
? SUDHIR KUMAR, D.P. SINGH and N.K. SINGH ......... 487
Varietal differences in fruit physico-chemical properties of strawberry (Fragaria x ananassa Duch.)
under mid hills condition of Himachal Pradesh
? SOURABH GARG, GIRISH SHARMA and MUKESH MEENA ........ 492
Identification and validation of informative SSR markers for genetic purity testing in hybrid rice (KRH-4)
? C. PUSHPA, RAME GOWDA, N. NETHRA and N. SHIVAKUMAR ........ 496
Multiplication & conservation of Rauvolfia serpentina L. - A threatened medicinal plant using tissue
culture techniques
? H.V. KALPANDE, V.S. PATIL, V.V. KALPANDE and MRIGANK SHEKHAR ........ 500
Genetic diversity analysis of soybean (Glycine max L.) cultivars revealed by PCR based molecular markers
? H.V. KALPANDE, A.W. MORE, J.D. DESHMUKH and A.N. NAIKNAWARE ........ 505

Quantitative analysis of lycopene content in selected F6 lines of tomato during post-harvest storage condition
? SUJEET KUMAR, P.H. RAMANJINI GOWDA, N.M. MALLIKARJUNA and SUKANTA SINHA ........ 509
Contd. ....
Effect of provenance and micronutrients on plant growth, seed yield and quality of maize hybrid (GH-0727)
? K.S. SIDDARUDH, M.R. ESHANNA, M.S. HARISH, N. GANGARAJU and S.R. SWAPNAKUMARI ........ 513
Effect of different levels of phosphorus on germination, drymatter production & sheath moisture
at different stages of sugarcane as influenced by cane trash & mycorrhizae
? T. USHA RANI, D. BALAGURAVAIAH and M. BHARATA LAXMI ........ 517
Effect of planting geometry and nutrient management in maize + mashbean intercropping system on
growth, productivity, nutrient removal, quality and nodulation
? ASHISH DWIVEDI, ADESH SINGH, R.K. NARESH, DINESH KUMAR, ROOP KISHORE and PREM NATH ........ 521
Integrated cultural management on the yield and yield traits of pigeonpea hybrids and varieties
? M.G. MULA, D.P. THAKARE, S.P. MEHTRE, A. RATHORE and ANILKUMAR VEMULA ........ 525
Yield & nutrient dynamics of organic nutrient management in scented rice under dates of transplanting &
seeding density followed by lentil (Lens esculenta)
? M.K. MANDAL, M. PRAMANICK, PINTOO BANDOPADHYAY and A. PHONGLOSA ........ 529
Response of seaweed saps on rice productivity (Oryza sativa L.)
? P.C. KANWAR, S.K. DWIVEDI, A. PAL and K.K. LODHI ........ 534
Influence of nutrient and mulch management practices on strawberry cv. Chandler under rainfed condition
? S.K. MAHATO, B. PARIYAR, BINOY CHHETRI and C.P. SURESH ........ 538

Yield, quality parameters & water use of summer sesame as influenced by irrigation scheduling & fertility levels
? R.B. THANKI, R.M. SOLANKI, J.M. MODHAWADIA, P.J. PRAJAPATI, B.S. GOHIL and N.B. BABARIA ........ 544

Effect of organic and integrated nutrient management on nitrogen and potassium dynamics, wheat yield
and nutrient uptake under Basmati-wheat sequence
? Z.A. BHAT, S.A. PADDER, G.S. SAROA, D.K. BENBI and O.P. CHOUDARY ........ 549

Screening of vigour status among aromatic rice through accelerated test

? VIKRAM JEET SINGH, SRIHIMA GAMPALA, S.K. CHAKRABORTI, G.R. MANJUNATH & K.P. VISHWAJITH ........ 555
Effect of different levels of phosphorus and sulphur on growth, yield and nutrient uptake of
semi-rabi pigeonpea (Cajanus cajan (L.) Millsp)

? MAHESH CHAND AGARWAL V.R. BHATT, M.B. VIRADIYA and B.S. KHERAWAT ........ 560
Response of biofertilisers and organic manures on growth, flowering, yield and quality of chrysanthemum
? NEELIMA PALAGANI, A.V. BARAD MANISHA SHINDE and NILIMA BHOSALE ........ 565
Effect of organic nutrient sources on forage yield in rainfed farming system
? S.K. SINGH, A. DEY and M.K. MEENA ........ 569
Effect of mulching on flowering, pod yield, water use & water use efficiency of French bean (Phaseolus vulgaris)
? B.V.G. PRASAD, S. CHAKRAVORTY, B.K. SAREN and D. PANDA ........ 573
Growth, yields, and nutrient uptake of finger millet (Eleusine coracana) as influenced by fertilizer
management through fertilizer briquettes
? A.V. BULBULE, P.N. GAJBHIYE and R.D. NIGADE ........ 576
A high yielding durum wheat variety UAS-428 released for cultivation to areas of Peninsular India
? SUMA S. BIRADAR, RUDRA V. NAIK, R.R. HANCHINAL, B.N. PATIL, I.K. KALAPPANAVAR
and S.A. DESAI ........ 580
Isolation & characterization of efficient exopolysaccharide producing bacteria from rhizosphere of wheat,
maize and mungbean crop grown in rainfed condition
? DEVENDRA SINGH, GEETA SINGH and K. ANNAPURNA ........ 583
Isolation and characterization of Gluconacetobacter sp. from fruit sources
? SUDHEERKUMAR TALAWAR, B. NARAYANASWAMY and S.N. RAVINDRA ........ 587

Evaluation of osmotolerant rhizobacteria for alleviation of water deficit stress in mustard

? BANDEPPA, SANGEETA PAUL and B.K. KANDPAL ........ 590
Contd. ....
Seed dormancy breaking methods in foxtail millet (Setaria italica) genotypes
? ANIL SEBASTIAN, S.N. VASUDEVAN, N.M. SHAKUNTALA, SANGEETA MACHA, R.P. PATIL and V.B. WALI ........ 594

Effect of some male gametocides on growth attributes in soybean

? S. ABIRAMI, A. KALAMANI and T. KALAIMAGAL ........ 598
Economic feasibility of intercropping of linseed (Linum usitatissimum L.) and chickpea under rainfed condition
? S.A. BIRADAR, K. AJITH KUMAR, RAJANNA B. and G.V. SHUBHA ........ 601
PCR detection of endosymbionts in whitefly collected from cassava and cassava based crops
? JITENDRA KUMAR, R. RABINDRAN and PREM KUMAR ........ 604
Influence of standardized feed additives on economic traits of mulberry silkworm hybrids CSR2 x CSR4 and
ND7 x CSR2 (Jayalakshmi)
? S.N. SUDHAKARA and NEELU NANGIA ........ 607

Biosafety studies against Chrysoperla eggs, larvae and ladybird beetle (Coccinellid) adults
? ASMITA SURADKAR and S.P. UKEY ........ 611
Evaluation for effectiveness of organic inputs for the management of Lipaphis erysimi Kalt. in mustard crop
? SURENDER KUMAR SHARMA, PUNAM, J.P. SAINI and RAKESH ........ 614
Dissipation dynamics of profenophos in polyhouse and open fields in tomato and evaluation methods for
removal of pesticide residues
? SUDHAKAR S. KELAGERI, CH. SREENIVASA RAO, I.B. BIRADAR, VASUDEVA NAIK K. and SUJATA ........ 617
Mass multiplication and shelf-life study of coconut water liquid formulation of Pseudomonas aeruginosa M.
? MANISHA S. SHINDE, A.N. SABALPARA, NEELIMA PALAGANI, R.R. WAGHUNDE and A.J. DESHMUKH ........ 622

Standardisation and preparation of accelerated fermentation of ‘idli’ batter using soy residue okara
? DEEPIKA G. KAMBLE and NITIN G. SURADKAR ........ 626
Suitability of drying methods for Roselle (Hibiscus sabdariffa L.) calyces
? R.R. ANDHALE, SYED IMRAN HASHMI, P.N. SATWADHAR and H.W. DESHPANDE ........ 630
Evaluation of cultivars and packing materials for quality of guava jelly bar during storage
? VENKATA SATISH KUCHI, RAVI GUPTA, RAJESH GUPTA and S.S. RATHORE ........ 634
Effect of post-harvest treatments on physical parameters changes in banana during ripening under
different storage conditions
? S.M. TALANE, B.V. PADHIAR, S.S. MINGIRE, A.Y. SAVVASHE and K.N. DHAWALE ........ 638
Effect of varieties & pre-treatments on physico-chemical composition of banana crisps
? K. VENKATA SUBBAIAH, S.L. JAGADEESH and R. MANJULA ........ 642

Effect of low fat-sugar free levels on sensory evaluation and economics of softy ice-cream
? A.S. CHAVAN, R.R. SHELKE, P.A. KAHATE and S.R. MUNNARWAR ........ 646
Rainfall analysis for crop planning in Thrissur district of Kerala
? B. AJITHKUMAR, G. SUBRAMANYAM and ARJUN VYSAKH ........ 650
Power generation using briquettes from de-oiled jatropha seed cake through gasification
? D. PADHEE, H. RAHEMAN, J.S. NIKHADE and B.L. SINHA ........ 653
Effect of income and prices on production and marketing behavior of maize farmer
? B. SOWJANYA, D.V. SUBBA RAO and R. VIJAYA KUMARI ........ 657
Performance evaluation of animal operated farm yard manure applicator
? M. CHOWDA REDDY and K.V. PRAKASH ........ 660

Strategic Vision : 27
Capacity development : A key ingredient towards sustainable agricultural growth
? ROSANA P. MULA Back Inner Page
 Green Farming Vol. 6 (3) : 622-625 ; May-June, 2015 Research Paper

Mass multiplication and shelf-life study of coconut water

liquid formulation of Pseudomonas aeruginosa Migula
a1* a2 b3 c4
MANISHA S. SHINDE , A.N. SABALPARA , NEELIMA PALAGANI , R.R. WAGHUNDE
d5
and A.J. DESHMUKH
a b
Deptt. of Plant Pathology, N. M. College of Agriculture, Deptt. of Floriculture and Landscape Architecture,
c
ASPEE College of Horticulture and Forestry, Bharuch College of Agriculture, Navsari Agricultural University,
d
Navsari - 396 450, College of Horticulture, S.D. Agricultural University, Sardarkrushinagar - 385 506 (Gujarat)

Received : 28 June 2014 ; Revised accepted : 02 April 2015

ABSTRACT
Seven media were tested for mass multiplication of Pseudomonas aeruginosa, among the natural media, coconut water
(143.66) induce maximum colony count. In sterile condition, among the different concentrations of coconut water tested, 30
per cent concentration showed significantly more population of P. aeruginosa (389). In unsterile condition among the
different concentrations of coconut water tested, 30 per cent concentration showed significantly higher population of P.
aeruginosa (455.33) as compared to rest. The shelf-life study done in conical flask and HDPE bottles, in conical flask at
fifteen days of storage of coconut water formulation, the population of P. aeruginosa at 10-7 dilution was recorded as 94.00. It
was found to increase upto 30 days and reached at it's highest (128.00). Then after it was observed to decrease upto 180
days (2.00). The population recorded at 180 days of storage (2.00) at room temperature (25±2oC). While in HDPE bottles, at
fifteen days of storage of coconut water formulation the population of P. aeruginosa at 10-7 dilution was reached at 99.00 then
increasing upto 30 days and reached at it's highest (125.00), then after it was observed to decrease upto180 days (1.00). The
population recorded (1.00) at 180 days of storage at room temperature (25±2oC). The highest cfu/ml was recorded at 30 days
of storage (1.25 x 1010 cfu/ml ) while the lowest (1 x 108 cfu/ml) at 180 days of storage.

Keyword : Coconut water, Mass multiplication, Natural media, Pseudomonas aeruginosa, Shelf-life.

INTRODUCTION
Liquid formulation based on chemical is available, but the
natural source based formulations are cheap and eco-friendly
which are not available. Development of liquid formulation which
ensures quality product in liquid over conventional carrier based
biofertilizers and biopesticides. These liquid formulations
facilitate long shelf-life up to 2 years, no need of carriers,
convenience of handling, storage and trans-portation, easy
quality control, more export potentials and also being preferred
by the community of farmers as well as manufacturers. In liquid
formulation, the microbial organism are present in a dormant
cyst form and after application in the field, the dormant form
gives to active cell. This help to increase the shelf life of liquid
formulation for more than 1 year (Vendan and Thangaraju,
2007). Recently, Kolombet et al., (2008) reported the extended
shelf-life of a Trichoderma asperellum in liquid formulation.
Coconut liquid endosperm, though nutritive in tender stages,
become depleted of nutrient as the nut matures. Mature coconut
when split open for making copra, the coconut water is
discarded as an agricultural waste which can be used as a good
medium for mass multiplication of bioagents (Jayalakshmy et
1,3 2 were transferred to each flask after cooling and incubate at
Ph.D. Scholar*(manisha5476@gmail.com), Director & Dean,
4,5
Asstt. Professor
al., 1988 and Nathanael, 1996). Hence, in present study cheap
natural media i.e. coconut water based liquid formulation of P.
aeruginosa was prepared.
MATERIALS AND METHODS
Natural source for mass multiplication of P. aeruginosa
in liquid state : Lab trial were conducted during 2012-2013 at
Biopesticide unit, Dept. of Plant Pathology, N.M. College of
Agriculture, N.A.U., Navsari. Lab trial were laid out in a CRD with
three replications and seven treatments.
Treatment detail : Sterilized conical flasks of 250 ml were
taken and 100 ml distilled sterilized water was filled in each
flasks. Ten ml of coconut water, jeevamruth, banana
pseudostem sap, cow dung extract and sugar cane juice were
prepared as described earlier which were added in each of the
above flasks, respectively. Further, these flasks were
autoclaved for 20 min at 121°C temperature and 15 lbs. Nutrient
broth and king's B broth were also kept for comparisons.
9
Inoculation of 0.1 ml of P. aeruginosa suspension (2 × 10 cfu/ml)
0 0
28 ±2 C temperature.

185
623 Shinde et al. Green Farming 6 (3)

Treatments : There were seven treatments (Table 1). water was added. The bottles were taken fitted with inner and
0
cap. These were sterilized in autoclave at 121 C temperature.
Observations : Colony forming units were counted after 48 9
Bacterial suspension (0.1 ml having 2 x 10 cfu/ml) of P.
hrs. of incubation. Samples were drawn after 48 hrs of
aeruginosa was added in the bottle using sterilized syringe
inoculation and the population of the P. aeruginosa were
aseptically. The bottles then were kept at room temperature for
counted, using viable plate counts on King's B Medium (KBM)
incubation. Four bottles were kept, which served as repetitions.
plates. For that 1ml of the suspension was taken and serial
dilution was made and 0.1 ml aliquot from 10-7 dilution were RESULTS AND DISCUSSION
spreaded on King's B medium and kept for incubation at room
Natural source for mass multiplication of P. aeruginosa
temperature (28±2 0C). Cfu/ml was calculated.
in liquid state : The study on the effect of different synthetic
To decide effective concentration of coconut water -7
and non-synthetic media used after 48 hrs. at 10 dilution
for mass multiplication of P. aeruginosa : In previous revealed that significantly higher colony count (494.00) was
trial, coconut water was found to be the best natural media for obtained in king's B broth as compared to rest. Next best
mass multiplication of P. aeruginosa. Moreover coconut water treatment in order of merit was nutrient broth (458.33). Among
is easily available and it is cheap source for natural mass the natural media, coconut water (143.66) induce maximum
multiplication media. Therefore coconut water was selected as a colony count and statistically at par with sugarcane juice
medium for mass multiplication of P. aeruginosa. This trial was (142.33) and cow dung extract (140.33). Jeevamruth (128.00)
taken under in two sets, i.e. under sterilized and non sterilized and banana pseudostem sap (73.66) were also good (Table 1).
conditions. Table 1. Effect of different media on population of
Sterilized condition : Sterilized conical flasks of 250 ml were P. aeruginosa
taken and 100 ml of distilled sterilized water was filled in each Population of Cfu/ml
Media
flask. Coconut water was added @ 10, 20, 30, 40 and 50 ml, in P. aeruginosa* after 48 hr.
the flasks treated as treatments. Further, these flasks were 10
Sugarcane juice (10 %) 142.33 1.42 x 10
sterilized in autoclave at 1210C for 15 min. After cooling, 0.1 ml of 9
Banana pseudostem sap (10 %) 73.66 7.37 x 10
P. aeruginosa suspension (2 × 109 cfu/ml) was transferred to 10
Jeevamruth (10 %) 128.00 1.28 x 10
each flask and they were incubated at 280±20C temperature. 10
Coconut water (10 %) 143.66 1.44 x 10
Treatment detail : There were seven treatments (Table 2). Cow dung extract (10 %) 140.33 1.40 x10
10

10
Unsterilized condition Kings B broth 494.00 4.94 x 10
10
Nutrient broth 458.33 4.58 x 10
Experimental detail : Fresh coconut water was added directly S. Em± 1.08
in the flasks with 100 ml distilled sterilized water. Other treatment CD at 5% 3.28
details and observations recorded are as per earlier trial. CV% 0.80
*Population of P. aeruginosa after 48 hr.10-7 dilution
Shelf-life study of coconut water formulation of P.
In all the synthetic as well as non-synthetic media tested
aeruginosa : An experiment was conducted to check the 9
produced a range of 73.66 to 494.00 population (7.37 x 10
shelf-life of coconut water formulation of P. aeruginosa during 10
cfu/ml to 4.94 x 10 cfu/ml) of the bacterium. This is above the
the year 2012-2013. Two sets were kept for shelf-life study i.e. in 8
standard of any liquid formulation count (1 x 10 cfu/ml) (Anon,
conical flasks and in HDPE bottles.
1968) for bioinoculant/biofertilizers as per the standards of ISI-
Conical flasks : Sterilized 150 ml flask were taken and filled Insecticide-1968 act. Coconut water, sugarcane juice, cow dung
with 100 ml distilled sterilized water. Fresh coconut water (30 ml) extract, jeevamruth or banana pseudostem sap can be used for
was then added in the flasks which were then plugged with non mass multiplication of the bacteria. Among these, coconut water
absorbent cotton, sterilized in autoclave at 1210C temperature. is the best natural source as it is the most effective, easily
Bacterial suspension (2 x 109 cfu/ml) of P. aeruginosa was added available and also cost effective..
(0.1 ml) by sterilized syringe aseptically. The flasks were The present study carried out here was for the first time in
incubated at room temperature for 6 months. Such 4 flasks were south Gujarat however the use of coconut water as a source of
kept, which served as repetitions. Total 15 sets were kept to take mass multiplication media for Pseudomonas spp. was in line
observations at 15 days interval. Samples were drawn after 15 with the work carried out by Anith (2009) at Vellayani and
days interval and the population of the P. aeruginosa were Mathew et al. (2010) at Kerala.
counted, using viable plate counts on King's B Medium (KBM)
plates. Mass multiplication of P. aeruginosa in coconut water :
In previous experiment, coconut water was proved as a better
HDPE bottle : Milky white virgin HDPE bottles (65±5 g) were natural liquid media for the mass multiplication of P. aeruginosa.
filled with 200 ml distilled sterilized water and 60ml fresh coconut Hence, the trial on testing of coconut water at different

186
May-June 2015 Mass multiplication & shelf-life of coconut water liquid formulation of P. aeruginosa 624

concentrations was carried out to decide application dose. The Table 3. Effect of different concentration of coconut water
trial was conducted in both, steriled and unsteriled conditions. on population of P. aeruginosa in unsterile condition
Concentration Population of Cfu/ml
Steriled condition : King's B broth resulted significantly
-7 (%) P. aeruginosa* after 48 hr.
higher population of P. aeruginosa at 10 dilution (495.33) as
10
compared to the rest. Next best treatment was nutrient broth 10 228.67 2.28 x 10
(451.67) in supporting the growth. 10
20 284.00 2.84 x 10
10
Among the different concentrations of coconut water tested, 30 455.33 4.55 x 10
10
30 per cent concentration showed significantly more population 40 370.00 3.70 x 10
of P. aeruginosa (389) as compared to the rest. Next best 10
50 327.00 3.27 x 10
treatment in order of merit was 40 per cent (354.33), the rest 10
Nutrient broth 455.00 4.55 x 10
concentration viz., 50 (297.33), 20 (201.33) and 10 per cent 10
Kings B broth 498.00 4.98 x 10
(182.67) also supported good growth. The highest colony
S. Em± 1.62
forming per ml (3.89 x 1010 cfu/ml) was recorded at 30 per cent CD at 5% 4.93
concentration while it was minimum (1.82 x 1010 cfu/ml) at 10 per CV% 0.75
cent. The minimum population recorded at 10 per cent is *Population of P. aeruginosa after 48 hr.10-7 dilution
sufficient as per ISI standard. Hence, the same may be 50 percent coconut water or 25 per cent coconut water
recommended. (Table 2). supplemented with sugar @15 g/l, as cheap nutritional liquid
Table 2. Effect of different concentrations of coconut medium for mass multiplication of Trichoderma harzianum and
water on population of P. aeruginosa in sterile condition Pseudomonas fluorescens. Mathew et al. (2010) also
suggested coconut water 50 per cent diluted in water for the
Concentration Population of Cfu/ml
(%) P. aeruginosa* After 48 hr.
mass multiplication of Pseudomonas fluorescens.

10 Shelf-life of coconut water formulation of P. aeruginosa

10 182.67 1.82 x 10
10 : At fifteen days of storage of coconut water formulation, the
20 201.33 2.01 x 10 -7
10 population of P. aeruginosa at 10 dilution was recorded as
30 389.33 3.89 x 10
10 94.00. It was found to increase upto 30 days and reached at it's
40 354.33 3.54 x 10
10 highest (128.00). Then after it was observed to decrease upto
50 297.33 2.97 x 10
10 180 days (2.00). During storage period, the colour of the
Nutrient broth 451.67 4.52 x 10 formulation was found to change. The colour of formulation
10
Kings B broth 495.33 4.95 x 10 when it was prepared originally it was milky white. It changed to
S. Em± 2.00 light green to milky white during the storage of 15 to 30 days.
CD at 5% 6.07
CV% 1.02 During 45 to 60 days of storage the colour was light to dark
*Population of P. aeruginosa after 48 hr.10-7 dilution green while during 90 to 105 days of storage it again changed to
pale yellow. During 105 to 150 days it was pale yellow and upto
Unsteriled condition : King's B broth resulted significantly
180 days colour was changed to pale yellow to light brown.
higher population of P. aeruginosa at 10-7 dilution (498.00) as
During 165 to 180 days, the colour of the suspension was pale
compared to the rest. Next best treatment was nutrient broth
yellow to light brown. The highest cfu/ml was recorded at 30
(455.00) in supporting the growth. 10 8
days of storage (1.28 x 10 cfu/ml ) while the lowest (2 x 10
Among the different concentrations of coconut water tested, cfu/ml) at 180 days of storage. The cfu recorded after 180 days
30 per cent concentration showed significantly higher of storage is sufficient as per ISI standard. (Table 4).
population of P. aeruginosa (455.33) as compared to rest. Next
best treatment in order of merit was 40 per cent (370.00). The Shelf life in HDPE bottle : At fifteen days of storage of
-7
rest concentrations viz., 50 per cent (327.00), 20 per cent coconut water formulation the population of P. aeruginosa at 10
(284.00) and 10 per cent (228.67) also supported good growth. dilution was reached at 99.00 then increasing upto 30 days and
The highest colony forming unit per ml (4.55 x 1010 cfu/ml) was reached at it's highest (125.00), then after it was observed to
recorded at 30 per cent concentration while it was minimum decrease upto180 days (1.00). The population recorded (1.00)
o
(2.28 x 1010 cfu/ml) at 10 per cent. The minimum population at 180 days of storage at room temperature (25±2 C). The
10
recorded at 10 per cent is sufficient as per ISI standard. Hence, highest cfu/ml was recorded at 30 days of storage (1.25 x 10
8
the same may be recommended (Table 3). cfu/ml ) while the lowest (1 x 10 cfu/ml) at 180 days of storage.
The results are more or less similar with the Anandraj and The cfu recorded after 180 days of storage is sufficient as per ISI
Sarma (1997) who developed formulation of Trichoderma standard. (Table 5).
harzianum and Gliocladium virens by using different Fermentation of bacterium inside the HDPE bottles were
concentrations of coconut water. Mathew et al. (2010) reported started after two days of incubation, which created the problem

187 Green Farming

625 Shinde et al. Green Farming 6 (3)

Table 4. Effect of storage period on the viability of o

(25±2 C) up to 6 months without hampering the quality. Hence,
coconut water formulation of P. aeruginosa at room coconut water can be used for commercial purpose. For this,
temperature up to six months in conical flask more experiments for confirmation is suggested.
Days of Av. No. of colonies Cfu Colour The results of present study are similar with Mugilan et al.
-7
storage at 10 dilution per ml change (2011) who studied on multiplication of P. striata in vermiculite,
9 lignite and nutrient broth and found higher population of P. stiata
15 94 9.4 x 10 Light green to 9
milky white
in nutrient broth (28.3 x 10 cfu/ml) upto 60 days. Ramanujam et
30 128 1.28 x 10
10
Light green to al. (2012) recorded shelf life of P. putida PBA-5 in liquid (using
milky white glycerol (2.5 %), polyethylene glycol (1.5 %) and yeast extract (1
10
45 101 1.01 x 10 Light green %) and talc formulation by observing cfu/ml at various intervals
9 1
60 84 8.4 x 10 Dark green and found that liquid formulation gave higher cfu (7.61 x 10
9 1
75 60 6.0 x 10 Light yellow cfu/ml) as compared to the talc formulation (4.98 x 10 cfu/ml) up
9
90 42 4.2 x 10 Light yellow to 6 months.
9
105 29 2.9 x 10 Pale yellow It was first attempt made in the Gujarat to develop coconut
9
120 21 2.1 x 10 Pale yellow water based liquid formulation of P. aeruginosa for the native
9
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