Available online at www.pacificjournals.

com
PACIFIC

P JOURNALS

Asiatic journal of BIOTECHNOLOGY RESOURCES

International Quarterly Journal of Bioscience

AJOBR

Asiatic Journal of Biotechnology Resources; 2011; 2(04)454-460

Short communication

Studies on microbial flora of fruit juices and cold drinks

Neha I. Bagde and P.M. Tumane
P.G.T.D. of Microbiology, Rastrasant Tukdoji Maharaj Nagpur University, Nagpur, Maharashtra, India *Correspondence: Neha I. Bagde, P.G.T.D. of Microbiology, Rastrasant Tukdoji Maharaj Nagpur University, Nagpur, Maharashtra, India, Contact No.9503486983, Email- pritneh@gmail.com Received 19 February 2011; accepted 16 March 2011 Available online: 31 March 2011

Keywords: Fruit juices, cold drinks, microorganisms E. coli, S. aureus

Abstract The good quality of food is sum of its physical and chemical quality. A total of three fruit juices like apple juice, pineapple juice, mosambi juice, which are commonly consumed two of carbonated and cold drinks, collected from local fruit juice stalls; of Nagpur city, were screened for various organisms present in them. In apple juice total bacterial count (TBC) was found between 20x105 to 1x105cfu/ml. Similarly in pineapple juice bacterial count were found between 15x105 to 1x10 5 cfu/ml and the mosambi juice has the bacterial count in the same range. In case of cold drinks due to various techniques which are well established were used for the process of bottling so there were very rare chances of finding a lot of bacterial count as in case of fruit juices, the bacterial count in case of cold drinks were found between 3x10 5 to 1x105cfu/ml. Mainly Staphylococcus aureus was detected in almost all the samples of fruit juices as well as of cold drinks. Escherichia coli obtained in all fruit juices but not in cold drinks. Many

Introduction In recent years the increasing consumer awareness has emphasized the need for microbiologically safe food. Since the human food supply consists basically of plants and animals or products derived from them, it is undesirable that our food supply can contain microorganisms in interaction with the food. When the micro-organisms involved are pathogenic, their association with our food is critical from a public health point of view. Serious health hazards due to presence of pathogenic microbes in food can lead to food poisoning outbreaks. In India the chance of transmissions of disease through fruit juices are due to unsatisfactory hygiene, adulteration practices. Micro-organisms are present both inside as well as outside of fruits and vegetable. At the time of consumptions the majorities of bacteria found on surface are usually Gram negative and belong to the Enterobacteriaceae. Many of those organism are usually non pathogenic to humans. The inner tissues of fruits are usually regarded as sterile. However, bacteria can be present in low number as a result of the uptake of water through certain irrigation or washing procedures.
Submitted online at submit@pacificjournals.com Asiatic Journal of Biotechnology Resources Asiatic J. Biotech Res. 2011; 2 (04)454-460 Punlished online at http://www.pacificjournals.com/ajobr Copyright 2011 Pacific Publishers International. All rights reserved. 0976-4992/AJOBR/$0.76-doi:03.2011/AJOBR-2011/02(04)/454-460

N. I. Bagde and P. M. Tumane

455

Cold drinks
Of the bottle beverages such as Limca, Coca-Cola, Mirinda etc. (Brand names) most of which are carbonate the commonly recognized flavors are cola, citrus flavors and sometimes herbal type such as root bears. Most of the beverages are prepared from extract or may be made from the juice and pulp of various fruits. All beverages are acidulated with edible acid such as citric, phosphoric, tartaric or lactic. They contain sugar, which imparts to them much of their flavors and their caloric value, although some dieters varieties do contain a non-caloric sweetener. Carmel and certified food color are used to enhance the appearance of the product. There is no evidence that carbonate beverages serve as a means for transmitting pathogenic organics. However, as thrust quenchers they should be free of undesirable organism in order to maintain a status as an adjacent to diet. Since the finished product will be have advantage, not only for public health control purpose, but also for the routine maintenance of plant sanitation. In the same way method of examination of ingredients will serve as an indicator of contamination and aid in the elimination of any undesired microorganism showing up in the finished product.
of these organisms can cause diseases in humans, which indicate that a large number of juices tested did not fulfill the guidelines for the microbiological quality of juices. It is therefore concluded that steps must be taken to improve the microbial quality of products. Copyright 2011 Pacific Publishers International. All rights reserved.

Fruit juices
Fruit juice and drinks are nutritious which offer great taste and health benefits. Most fruit juices and drink brought from grocery and supermarket shelves are pasteurized. However a small percentage of juices are unpasteurized. This means that there is a chance the product may contain bacteria harmful to our health. Unpasteurized fruit and drinks have posed serious health risks in recent years. Worldwide, health concern has led to the popularization of natural fruit juice as a healthy alternative to other beverages. Fruit juices and drinks are nutritious which offer great taste and health benefits. (Suaad and Eman, 2008) The spoilage of food is most after due to contamination with aerobic acid-tolerant bacteria as well as yeasts and molds. Thus, enumeration of these microorganisms is an important aspect of evaluating the microbiolog y quality of acidic foods. Fruit juice are the unfermented, but fermentable, liquid obtained from the edible part of sound appropriately mature and fresh fruits or fruits maintained in fresh condition by physical mean or other suitable treatments. Juices can be obtained by mechanical extractions process or by reconstitution of concentrated fruit juice with potable water. Juices can be cloudy or clear and must have the essential characteristics typical of the juice of the fruit from which it comes. Diluting and
Copyright 2010 Pacific Publishers Asiatic J. Biotech Res. 2011; 2(04)454-460

456

N. I. Bagde and P. M. Tumane

Figure 1. Colonies of S.aureus on MSA.

Figure 2. Colonies of E.coli on EMB Agar.

or blending are a common practice as many fruit juices are either too acid or too strongly flavored to be pleasant for consumption. Fruit juice drinks typically contain no less than 20% fruit juice.

Concentrated fruit juice

It is the product that complies with the definition of fruitjuice, except that water has been physically removed, e.g. by evaporation. In the production of juice that is to be concentrated appropriate technological processes are used and may include simultaneous diffusion of the pulp, cells, or fruit pulp by water provided that the water extracted juice is added in line to the primary juice, before the concentration process. Two or more kind of fruit juice concentrates can be mixed together. Fruit juices concentrates may have added or restored aromatic substances, volatile flavor components, pulp and cells, all of which must be recovered from the same kind of fruit and be obtained by physical means. The microbiology of fruit juice concentrates is greatly influenced by their reduced water activity. Freshly squeezed fruit and vegetable juices have little or no process step that reduce pathogen level, if contaminated, such as no kill step. Freshly squeezed juices are simply prepared by extracting, usually mechanical means, the liquid and pulp of mature fruit or vegetable. The final product is an unfermented, unclarified, untreated juice, ready for consumption. During the process, contamination from raw materials, equipment or food handlers could be easily transferred to final product. If pathogens such as Salmonellae were present in freshly squeezed juice individuals may be exposed. Pathogenic organisms can enter fruits and vegetables through damaged surfaces, such as punctures, wounds, cuts and split. This damage can occur during maturation or during harvesting
Asiatic J. Biotech Res. 2011; 2(04)454-460 Copyright 2010 Pacific Publishers

N. I. Bagde and P. M. Tumane

457

and processing. A pathogen that has become internalized with in a fruit or vegetable must be able to survive in the product until it reaches the consumer in order to become a public health hazards. Most fruit juice is sufficiently acidic to inhibit the growth of pathogenic organisms. Studies conducted on the internalization survival, or growth of micro-organism can be present and survive in a wide range of fruit.

Sources of contamination
Fruit & fruit juices are commonly contaminated with bacteria, often from insect damaged. Fallen fruit thus be avoided were possible for all of the risks. Flavoring, water and other chemical are all potential sources of microbial contamination. Bacterial that have been associated with spoilage in soft drink industries include Acetobacter, Bacillus, Clostridium, Gluconobacter, Lactobacillus, Leccuonostoc, Saccharobacter, Zymobacter and Zymomonas . Gluconobacter is common spoilage agent of fruit juices. Quality control

The general purpose of quality control is to ensure that a maximum amount of the product being process reaches the desired level of quality with minimum variation and that this is achieved as economically as possible. The products of natural raw material are never exactly the same, so control is necessary to keep product quality within the standards set raw materials showed be purchased from reliable supplies who hold a current food manufactures registration. Quality control generally involves inspections of three kind of material such as 1. Raw Materials, 2. Materials in process and 3. Finished product. If effective raw material and process controls are not put in place and only examination of the finished product is done, then quality control stops and becomes merely an inspection .A good control system rejects substandard ingredients before the process begins and once, it has begun, prevents wastage of good raw material.

Materials and methods

Sample collection and processing Fruit juice samples were collected in sterile screw cap containers from local market of Nagpur city. All samples were homogeneously mixed before the microbial examination.Table1 Diluted serially and aliquots of the appropriate dilutions were then plated on nutrient agar plate and on different selective media for the isolation of
Copyright 2010 Pacific Publishers Asiatic J. Biotech Res. 2011; 2(04)454-460

458

N. I. Bagde and P. M. Tumane

Table 1. Representing sample categories with types.

Sample category Fruit juices

Specific types Apple juice pineapple juice Mosambi juice Coke Orange flavor (2) (2) (2) (2) (2)

Cold drinks

Table 2. Enumeration of bacteria from fruit juice sample.

Bacterial count x 10 5cfu/ml Samples Apple juice Pineapple juice Mosambi juice Cold drink 1 Cold drink 2 Minimum 2x10 5 1x10 5 1x10 5 1x10 5 2x10 5 Maximum 20x105 18x105 15x105 3x105 5x105 Mean 11x105 9x105 8x105 2x105 4x105

food borne microorganisms to assay their colony-forming unit. The plates were incubated at 370C for 24 hrs. Identification of organism Morphological and biochemical characteristics Identification of organisms was carried on basis of Morphological, Biochemical and cultural characteristics. Bacterial isolates were analysed for Gram character and their motility and Various biochemical tests were performed by inoculating small portion of well-isolated colony into a series of media and performed Sugar Fermentation, Indole Test, Methyl Red Test, Voges Proskauer Test, Triple Sugar Iron Agar, Urease Test, Citrate Utilization Test, Catalase Test. Cultural characteristic Additionally many tests were performed to assess the biochemical or metabolic activities, specific to each bacterial isolate by inoculating series of a respective differential test media prepared as per the standard procedure by observing their culture characteristic on MacConkey agar, Baired Parker Agar, Eosin Methylene Blue Agar, Bismuth sulphite Agar (BSA), Xylose- Lysine Deoxycholate Agar (XLD), Deoxycholate Citrate Agar (DCA) [Himedia 1998].
Asiatic J. Biotech Res. 2011; 2(04)454-460 Copyright 2010 Pacific Publishers

N. I. Bagde and P. M. Tumane

459

Result and discussion

Isolation of micro organisms from fruit juices From the experiment carried out, it was observed that from apple juice 3 bacterial isolates were obtained and they were maintained on nutrient Agar slants. Similarly from pineapple juice 2 bacterial isolates were obtained and from mosambi Juice 5 bacterial isolates were obtained. They were maintained on nutrient agar slant. Total bacterial counts of sample examined ranged between 20x10 5to 1x10 5 in cases of fruit juices. In case of cold drinks total bacterial count were found in the range between 3x10 5 to1x10 5.Staphylococcus aureus was detected in almost all the samples of fruit juices as well as of cold drinks. Escherichia coli obtained in all fruit juices but not in cold drinks while in one, Salmonella typhi in one. The objective of present study was to isolate microorganism present in natural fruit juice and cold drinks. Among isolates three were obtained from apple juice, five isolates from mosambi juice, and two isolates from pineapple juice and four from cold drinks. The need for microbiological examination in this field can be found in public health aspects of production, distribution and wide spread use of beverage products and the need for determination of the causes and prevention of spoilage. Bottled beverages are consumed as thirst quenchers. As much they should meet or be better than the requirement for drinking water as regards the various types of organisms. Whenever, the finished beverages do not meet the requirements for drinking water, according to Public Health Service Drinking Water Standards, there is definite indication of unsatisfactory raw material or very faulty processing methods. In most cases the primary source and carrier for coliform organism is the water supply employed, but in rare instances they have been found human contamination sources within the processing plant which lead to excessively high counts of the organism. High mold or bacteria counts may be indicative of poorly cleaned equipment on reflective of the condition of fruit used and high yeast or lactic acid forming bacteria may be an indication of fermentation of the product. The use of good quality raw fruit is essential to the production of a high quality product of low microbial count. Wolford and Berry (1948) have shown that juices produced from soft rot fruit contains many times the total bacteria number found in juice from sound fruit. Faville and Hill (1952) found that strains of species of the genera Leuconostoc, Aerobacter, Lactobacillus, Xanthomonas and Achromobacter may grow in raw fruit and that excessively high counts in juice may indicate indequent inspection and incomplete elimination of unsound fruit.
Copyright 2010 Pacific Publishers Asiatic J. Biotech Res. 2011; 2(04)454-460

460

N. I. Bagde and P. M. Tumane

Brokaw (1953)has pointed out the vital role of plant sanitation in maintaining quality in juice and concentrate production. The study carried out by Joy E-Lewis et. al (2006)on the street vended fruit juices in Vishakhapatnam city, India, indicates that all street vended fruit juices showed contamination with fecal coliforms and faecal streptococci. It is contended that contamination is mainly due to poor quality of water used in dilution as well as prevailing unhygienic conditions related to washing of utensils and maintenance of premises.

References
1. Brokaw, C. H. (1953) The role of sanitization in quality control of citrus Concentrates. Food Engr, 25 (7); 94-95. 2. Faville L. W. & Hill E.C. (1952) Acid tolerant bacteria in Citrus Juice Ibid, 17:281-287. 3. Lewis Joy .E., C. Kalavati, B. Rajanna.(2006) A case study in Vishakhapatnam city, India. Journal of Food Safety, 8, 35-38. 4. Suaads, Alwakeel and Eman Abdullah Hamed (2008). Microbial growth and chemical analysis of Botteled fruit juices and drinks in Riyadh, Saudi Arabia. Research J of Microbiology, 3, 315-325. 5. Wolford E.R. & Berry J. A. (1948). Condition of Oranges as affecting bacterial content of frozen juice with emphasis on coliform organism. Food research,13: 172-178.

Asiatic J. Biotech Res. 2011; 2(04)454-460

Copyright 2010 Pacific Publishers