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ASIAN J. EXP. BIOL. SCI.

VOL 4(4) 2013: 651-653

© Society of Applied Sciences

ORIGINAL ARTICLE

Morphological, Physiological and Biochemical Characterization of
Bacteria Isolated From Pineapple Juice
Neetu Singh, Rajesh Kumar, Sunita Khatak, Deepak Kumar Malik and Anita Grewal
Department of Biotechnology Engineering, University Institute of Engineering &Technology,Kurukshetra University
Kurukshetra-136119, Haryana
Received 09-09-2013
Revised 30-09-2013
Accepted 20-10-2013
ABSTRACT
In the present study eight bacterial cultures (NT1, NT2, NT4, NT5, NT6, NT7, NT8 and NT9) were isolated from pine apple
juice. Out of eight isolates only two bacterial cultures (NT6 and NT7) were gram positive. Rest of six isolates was gram
negative. The physiological characteristic of bacterial isolates was checked by their growth at different temperature, pH and
salt concentration. The bacterial isolated (NT1, NT2 and NT4) on the basis of their morphological, physiological
biochemical characteristics tentatively characterized as vibrio sp.
Keywords: Biochemical characterization, pine apple,

INTRODUCTION
Pineapple is considered as the second most economically important tropical fruit. Pineapple is the only source of
complex proteolytic enzyme used in the pharmaceutical market and as a meat-tenderizing agent. Pineapple fibre has
been processed into paper with remarkable qualities of thinness, smoothness and pliability [1]. The use of pineapple
for colds as well as for asthma is also well known. Alcoholic beverages can also be made from juice. Fruit and fruit
juices are commonly contaminated with bacteria, often from insect damaged. Flavoring, water and other chemicals
are all potential sources of microbial contamination. Gluconobacter is common spoilage agent of fruit juices.
Alicyclobacillus spoilage has become an important issue in heat-treated fruit juices, and the bacterium is commonly
found contaminating fruit in the field. Growth of the organism is associated with the production of antiseptic and
'smoky' taints within juice. There are few reports on the diversity of microbial inhabitants of pineapple plants. This
plant is colonized by endophytes like Gluconacetobacter diazotrophicus [2], Burkholderia sp. [3], related to
Azospirillum [4]. Keeping in view of some medicinal properties of pineapple juice our objective was to identify the
bacterial community of pine apple juice.

MATERIALSAND METHODS
Isolation of bacteria from pineapple juice
The dehydrated media and biochemical test kit were procured from Hi Media. The bacteria were isolated from pine
apple juice by spread plate method on nutrient agar plates. The isolated bacterial culture was maintained on nutrient
agar slants for further study. The Gram staining of isolated culture were performed by using Hi media staining kit
Physiological and biochemical characterization of isolated bacterial culture
The isolated bacterial strain were grown over nutrient agar plates at different temperature (4 ºC, 10 ºC, 20 ºC, 25 ºC, 30
ºC, 35 ºC, 40 ºC, 50 ºC, 60 ºC and 70 ºC), different pH (4, 6, 8 and 10) and different NaCl concentrations (4%, 6%, 8%
and 10%). The plates were incubated at 37 ºC for 72 hours and results were recorded. The biochemical
characterization of isolated culture was done by using Hi media identification kit.

ASIAN J. EXP. BIOL. SCI. VOL 4 (4) 2013

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8 and 10 % NaCl All the culture were not growing All the culture were growing Culture NT4. The growth of isolated cultre at different pH is also shown in Table 2. 10 °C. NT2. EXP. pH and% NaCl Conditions Growth 4°C. 50°C. 6 . SCI. All isolated culture was not grown at temperature (4 °C. Biochemical characteristics of isolated bacterial strains Biochemical tests Citrate utilization Lysine and ornithine utilization Urease and H2S production Phenylalanine deamination Nitrate reduction 652 Culture NT1 + - Culture NT2 - Culture NT4 + + + Culture NT5 + + - Culture NT8 + + - Culture NT9 + + - ASIAN J. NT5. 25°C. The biochemical characstics (citrate utilization. nitrate reduction and H2S production) of bacterial culture (NT1. The characterization of bacteria with biochemical test kit is tentative. adonitol and sorbitol) is shown in Table 4. He reported that cocci species constitute the major contaminants in the juices. NT7. The morphological characteristics of all the isolated culture is shown in Table 1. arabinose. NT6 and NT9 were growing Table 3. BIOL. NT4. 10°C. Morphological characteristics of isolated bacterial strains Colony charactertics Configuration Margin Elevation Surface Colour Opacity Gram’s reaction Cell shape Culture NT1 Circular Entire Elevated Rough Yellow Opaque Rod Culture NT2 Circular Entire Flat Smooth Yellow Opaque Rod Culture NT4 Circular Entire Flat Smooth Cream Opaque Rod Culture NT5 Circular Entire Elevated Rough Whitish Opaque Coccus Culture NT6 Circular Entire Elevated Smooth Yellow Transparent + Coccus Culture NT7 Circular Entire Flat Smooth Red Opaque + Coccus Culture NT8 Circular Entire Flat Smooth Orange Transparent Coccus Culture NT9 Irregular Entire Elevated rough Lemon Transparent Coccus Table 2. NT6. Morphological. pH10 4. 50 °C and 70 °C) and salt concentration (4. NT5.Neetu Singh et al. pH8. NT8 and NT9) were isolated on nutrient agar plate by using spread plate method. ornithine utilization. lactose. urease.. Growth of isolated bacterial strains at different temperature. NT8 and NT9) is shown in Table 3 and charbohydrate metabolism (glucose. Physiological and Biochemical Characterization of Bacteria Isolated From Pineapple Juice RESULTSAND DISCUSSION The 8 different type of bacterial culture (NT1. NT2. lysine. VOL 4 (4) 2013 . Table 1. On the basis of these biochemical tests bacterial culture NT1. The isolation of microorganism from natural pineapple juice was performed [5]. 8 and 10 % NaCl) as shown in Table 2. 6 . NT2 and NT4 was tentatively identified as vibrio sp. NT4. The need for microbiological examination in this field can be found in public health aspects. 35°C and pH6. distribution and wide spread use of juice. NT5. phenylalanine deamination. 70 °C.

. G. EXP. Philippines. Teixeira. Physiological and Biochemical Characterization of Bacteria Isolated From Pineapple Juice 4.com ASIAN J. V. Weber. Microb.. Olubukola. B.. L. Salgado. Appl. Pina.M.O. Environ. Department of Biotechnology Engineering. Ecol. (2000). Cruz. Baldani. K. Souza. Corresponding Author: Anita Grewal. (2011). Baldani. Baldani.B.I. O. 16S ribosomal DNA characterization of nitrogen-fixing bacteria isolated from banana and pineapple. J. Natural endophytic occurrence of Acetobacter diazotrophicus in pineapple.B. E.. F. O. Mellado.. J. Cristales. NT2 and NT4 was tentatively identified as vibrio sp. physiological and biochemical characstics bacterial culture NT1... Life Science Journal. Kirchhoff. E. Morphological. A. M. Carbohydrate metabolism of isolated bacterial strains Biochemical tests Glucose and arabinose Lactose and sorbitol Adonitol Culture NT1 - Culture NT2 - Culture NT4 + - Culture NT5 + Culture NT8 - Culture NT9 - CONCLUSION The 8 different type of bacterial culture were isolated form pineapple juice by using spread plate method. Plant Soil.E. L.M. Obashola. and Ramokoni. J. E. On the basis morphological.Neetu Singh et al. Microbiol. 67: 2375–2379. The need for microbiological examination can be found in public health aspects. and Ramírez..R. and Pedrosa.D.Amon Foundation.I. Isolation and characterization of diazotrophic bacteria from banana and pineapple. J. H a r y a n a .1 3 6 1 1 9 . 8: 18-24. T. Email: anitapunia_17@rediffmail. J...F. Microbiological quality control study of some processed fruit juices b y conventional approach. 39: 49–55. BIOL.R.G. University Institute of E n g i n e e r i n g & Te c h n o l o g y. Manila.T.S... Weber. SCI. L. (1999). 210: 103–113. (1991).L. K u r u k s h e t r a U n i v e r s i t y K u r u k s h e t r a .. Dbereiner. Dobereiner.. VOL 4 (4) 2013 653 . distribution and wide spread use of juice.O. REFERENCES [1] [2] [3] [4] [5] Montinola. Hernandez.. (2001).R.