Isolation and identification of microflora of farm ponds and gastrointestinal tract of western white shrimp (Litopenaeus vannamei) beside their evaluation as probiotic
Subject Areas : Molecular MicrobiologyMina Ziarati 1 , Mehran Avakh Keysami 2 , Farshid Kafilzadeh 3
1 - Department of Microbiology, Jahrom Branch, Islamic Azad University, Jahrom, Iran.
2 - Department of Aquaculture Biotechnology, Bushehr Agricultural Training Center, Bushehr, Iran.
3 - Department of Microbiology, Jahrom Branch, Islamic Azad University, Jahrom, Iran.
Keywords: Gastrointestinal tract, Litopenaeus vannamei, Pathogenic bacteria,
Abstract :
Background and Objectives: Nowadays, Shrimp culture has been prevailed in most of tropical countries; however, a variety of diseases have currently affected the business worldwide. Infectious bacterial agents are the most significant hygienic problems observed in the cultured shrimps . This study was aimed to isolate and identify microflora of farm ponds and western white shrimp gastrointestinal tract (Litopenaeus vannamei) in the Bushehr Province farm ponds and their survey as probiotic. Materials and Methods: The shrimp and pond water samples were collected from 10 farm ponds (located at Bushehr Province). After transferring the samples to laboratory, the shrimp samples were autopsied to access to their intestinal microflora. The isolated bacteria were identified based on biochemical tests, morphology and Biolog software. Antibacterial activities of the isolates against 6 common pathogenic bacteria in shrimps were then investigated using well diffusion and disk diffusion methods. Results: Overall, 12 bacterial genera were isolated from the farm ponds and shrimp gastrointestinal tract. The majority of bacterial species most abundantly isolated were Bacillus, Vibrio and Pseudomonas. Among the isolated bacteria, Bacillus, Micrococcus and Corynebacterium showed the most antibacterial activity against evaluated pathogenic bacteria. Conclusion: Almost all the isolated bacteria from shrimp gastrointestinal tract were found in farm ponds as well. The bacterium Bacillus sp., for instance, is able to inhibit the growth of pathogens through secreting antimicrobial products.
1. Barman P, Banerjee A, Bandyopadhyay P, Mondal KC, Das Mohapatra PK. Isolation, identification and molecular characterization of potential probiotic bacterium, Bacillus subtilis PPP 13 from Penaeus monodon. Biotechnol Bioinf Bioeng. 2011; 1(4): 473-482.
2. Lightner DV. Red disease of penaeid shrimp. In: Sindermann CJ, Lightner DV (eds.). Disease diagnosis and control in north American marine aquaculture. Developments in aquaculture and Fisheries science 17. Elsevier, Amsterdam. 1988; pp. 100-103.
3. Nash G, Charetana N, Cholada T, Anutra A, Phusit P, Pongcham R. Vibriosis and its control in pond-reared Penaeus monodon in Thailand In: Diseases in Asian aquaculture (Eds: Shariff M, Subasinghe R, Arthur JR). Fish Health section, Asian Fisheries society, Manila. Phillippines. 1992; pp. 143-145.
4. Keysami MA, Saad CR, Daud HM, Sijam K, Alimon AR. Effect of Probiotic on larval growth and development rate of Macrobrachium rosenbergii (deMan). Kustem 4th Annual Seminar held at Primula Beach Resort, Kuala Terengganu, Malaysia during May 2-3, 2005. Abstract book, p: 80.
5. Keysami MA, Saad CR, Daud HM, Sijam K, Alimon AR. Effects of putative probiotic bacterium on the survival and growth of juvenile Macrobrachium rosenbergii (deMan). The Asian Aquafeeds 2005 Seminar which is scheduled for April 12-13, 2005, in Kuala Lumpur, Malaysia. Abstract book, p: 44.
6. Zokaeifar H, Balcazar JL, Kamarudin MS, Sijam K, Arshad A, Saad CR. Selection and identification of non-pathogenic bacteria isolated from fermented pickles with antagonistic properties against two shrimp pathogens. J Antibiot. 2012; 65(6): 289-294.
7. Iranian Shrimp Research Center. Available at http://www.isrc.ir
8. MacFaddin JF. Biochemical tests for identification of medical bacteria. 2nd edn. Williams and Wilkins, Baltimore, MD. 1980; p: 527.
9. Keysami MA, Saad CR, Daud HM, Sijam K, Alimon AR. To screen three putative bacteria in juvenile Macrobrachium rosenbergii as probiotic based on in-vitro growth characteristics. MSA Golden Jubilee International Science Congress (ISC). August 3-6, 2005 Putra World Trade Centre, Kuala Lumpur, Malaysia. Abstract book, pp: 144-145.
10. Keysami MA, Saad CR, Daud HM, Sijam K, Alimon AR. A preliminary study to isolation and identification of the aerobic putative bacterial flora in juvenile giant freshwater prawn, Macrobrachium rosenbergii (de Man). Kustem 4th Annual Seminar held at Primula Beach Resort, Kuala Terengganu, Malaysia during May 2-3, 2005. Abstract book: 78.
11. Chythanya R, Karunasagar I. Inhibition of shrimp pathogenic Vibrios by marine Pseudomonas 1-2 strain. Aquacult. 2002; 208(1-2): 1-10.
12. Rattanachuay P, Kantachote D, Suntinanalert P. Selection of proteolytic bacteria with ability to inhibit Vibrio harveyi during white shrimp (Litopenaeus vannamei) cultivation. Technol. 2007; 29(2): 235-243
13. Keysami MA, Saad CR, Daud HM, Sijam K, Alimon AR. Screening putative bacteria of Macrobrachium rosenbergii juvenile as potential applications for control of pathogenic bacteria. Kustem 4th Annual Seminar held at Primula Beach Resort, Kuala Terengganu, Malaysia during May 2-3, 2005. Abstract book: 122.
14. Phatarpekar PV, KenkreVD, Sreepada RA, Desai UM, Achuthakutty CT. Bacterial flora associated with larval rearing of the giant freshwater prawn, Macrobrachium rosenbergii. Aquacult. 2002; 203(3-4): 279-291.
15. Miyamoto G, Brock J, Nakamura R, Nakagawa L, Shimojo R, Sato V, Akita G. A preliminary microbiological and water quality survey of two Hawaiian prawn (Macrobrachium rosenbergii) hatcheries. In: Rogers GL, Day R, Lim A (Editors), Proc. First Int. Conf. Warm Water Aquaculture-Crustacea. Brigham Young University Hawaii Campus, Laie, HI. 1983; pp. 429-458.
16. Anderson IG, Shamsudin MN, Nash G. A preliminary study on the aerobic heterotrophic bacterial flora in giant freshwater prawn, Macrobrachium rosenbergii, hatcheries in Malaysia. Aquacult. 1989; 81(3-4): 213-223.
17. Sahul Hameed AS. A study of the aerobic heterotrophic bacterial flora of hatchery-reared eggs, larvae and post-larvae of Penaeus indicus. Aquacult 117(3-4): 1993; 195-204.
18. Llobrera AT, Gacutan RQ. Bacteria from sea water used in Penaeus monodon larval cultures. Quarterly Research Report, Aquaculture Department, SEAFDEC. 1977; 1(2): 38-40.
19. Keysami MA, Saad CR, Daud HM, Sijam K, Alimon AR. Isolation and identification of putative bacterial flora in juvenile Macrobrachium rosenbergii. MSA Golden Jubilee International Science Congress (ISC), Putra World Trade Centre, Kuala Lumpur, Malaysia, August 3-6, 2005. Abstract book: 92.
20. Janarthanam K, George MR, John KR, Jeyaseelan MJP. In vivo and in vitro biocontrol of vibrio using indigenous bacterium Bacillus spp. Indian J Geomarine Sci. 2012; 41(1): 83-89.
21. Liu CH, Cheng W, Hsu JP, Chen JC. Vibrio alginolyticus infection in the white shrimp Litopenaeus vannamei confirmed by polymerase chain reaction and 16s rDNA sequencing. Dis Aquat Organ. 2004; 61: 169-174.
22. Sung HH, Hsu SF, Chen CK, Ting YY, Chao WL. Relationships between disease outbreak in cultured tiger shrimp and composition of vibrio communities in pond water and shrimp hepatopancreas during cultivation. Aquacult. 2001; 192(2): 101-110.
23. Song YL, Cheng W, Wang CH. Isolation and characterization of Vibrio damsela infection form cultured shrimp in Taiwan. J Invertebr Pathol. 1993; 61: 24-31.
24. Venkat HK, Sahu NP, Jain KK. Effect of feeding Lactobacillus-based probiotics on the gut microflora,growth and survival of postlarva of Macrobrachium rosenbergii(de Man). Aquacult Res. 2004; 35(5): 501-507.
25. Bengmark S. Ecological control of the gastrointestinal tract. The role of probiotic flora. Gut. 1998; 42(1): 2-7.
26. Rengpipat S, Rukpratanporn S. Piyatiratitivorakul S, Menasaveta P. Immunity enhancement in black tiger shrimp (penaeus mondon )by a probiont bacterium (Bacillus S11). Aquacult. 2000; 191: 271-288.
27. Gomez-Gil B, Roque A, Turnbull JF. The use and selection of probiotic bacteria for use in the culture of larval aquatic organism. Aquacult. 2000; 191(1): 259-270.
28. Zokaeifar H, Balcázar JL, Saad CR, Kamarudin MS, Sijam K, Arshad A, Nejat N. Effects of Bacillus subtilis on the growth performance, digestive enzymes, immune gene expression and disease resistance of white shrimp, Litopenaeus vannamei. Fish Shellfish Immunol. 2012; 33(4): 683-689.
29. Zimmerman SB, Schwartz CD, Monaghan RL, Pelak BA, Weissberger B, Gilfillan EC, Mochales S, Hernandez S, Currie SA, Tejera E. Difficidin and oxydifficidin: novel broad spectrum antibacterial antibiotics produced by Bacillus subtilis. I. Production, taxonomy and antibacterial activity. J Antibiot. 1987; 40(12): 1677-1681.
30. Alavandi SV, Vijayan KK, Santiago TC, Poornima M, Jithendran KP, Ali SA, Rajan JJ. Evaluation of Pseudomonas sp. PM 11 and Vibrio fluvialis PM 17 on immune indices of tiger shrimp, Penaeus monodon. Fish Shellfish Immunol. 2004; 17(2): 115-120.
31. Vijayan KK, Bright Singh IS, Jayaprakash NS, Alvandi SV, Pai SS, Preetha R, Rajan JJS, Santiago TC. A brackishwater isolate of Pseudomonas PS-102, a potential antagonistic bacterium against pathogenic vibrios in penaeid and non-penaeid rearing systems. Aquacult. 2006; 251(2-4): 192-200.