جداسازی و غربالگری باکتریوفاژهای دریایی به منظور کاربرد در پیشگیری از بروز بیماری ویبریوزیس در میگوی سفید غربی: DOR: 20.1001.1.17359880.1400.14.3.4.5
محورهای موضوعی : مجله پلاسما و نشانگرهای زیستیسمیرا زارعی 1 , نیما بهادر 2 , مریم میربخش 3 , محمد خلیل پذیر 4
1 - گروه میکروب شناسی، دانشکده علوم، کشاورزی و فناوری‌های نوین، واحد شیراز، دانشگاه آزاد اسلامی، شیراز
2 - گروه میکروب شناسی، دانشکده علوم، کشاورزی و فناوری‌های نوین، واحد شیراز، دانشگاه آزاد اسلامی، شیراز
3 - مؤسسه تحقیقات علوم شیلاتی کشور، سازمان تحقیقات، آموزش و ترویج کشاورزی، تهران، ایران
4 - پژوهشکده میگوی کشور، موسسه تحقیقات علوم شیلاتی کشور، سازمان تحقیقات، آموزش و ترویج کشاورزی، بوشهر، ایران
کلید واژه: میگوی سفید غربی, جداسازی و غربالگری, باکتریوفاژ, بیماری ویبریوزیس,
چکیده مقاله :
زمینه و هدف: یکی از بزرگ ترین تهدیداتی که صنعت آبزی‌پروری با آن مواجه است، بیماری‌های عفونی باکتریایی و استفاده از آنتی بیوتیک ها است. فاژدرمانی یک جایگزین قابل اعتماد برای آنتی‌بیوتیک‌ها برای غیرفعال‌کردن باکتری به عنوان عوامل اصلی بیماری‌زا در صنعت آبزی‌پروری می باشد. هدف از تحقیق حاضر غربالگری گونه های ویبریو در سواحل استان بوشهر و کنترل این عامل بیماری زای میگو با استفاده از فاژ بود.روش کار: جهت انجام این مطالعه آب در سال 1398 و از مناطق نزدیک به سواحل استان بوشهر نمونه برداری شد. باکتری های عامل ویبریوزیس با استفاده از آزمایش های بیوشیمیایی و توالی یابی مولکولی تعیین شد. انتخاب باکتری هدف با توجه به نتایج پروفایل آنتی بیوگرام باکتری های غالب و پاتوژن ها انجام گردید. جهت تعیین تیتر فاژ محیط کشت TCBS به کار برده شد و شمارش پلاگ ها 24 ساعت پس از گرمخانه گذاری در دمای 30 درجه سانتی گراد انجام گرفت.یافته ها: از آب های آلوده به فاضلاب بوشهر، 5 باکتری(Vibrio alginolyticus strain SeqID، Vibrio xuii strain Z-03Phage، Vibrio alginolyticus SZ/10 gene، Vibrio rotiferianus strain SRPR-Phage، Vibrio harveyi strain Z05) جداسازی شد. با توجه به نتایج تعیین پروفایل آنتی بیوگرام، قطر هاله ی عدم رشد و آزمون بیوشیمایی باکتری گونه های مورد مطالعه، باکتری Vibrio alginolyticus strain SeqID به عنوان گونه ی باکتری غالب انتخاب شد. بر اساس نتایج حاصل از آنالیز مولکولی 16srDNA سویه جداسازی شده باکتری با 100% شباهت با باکتری ویبریو آلجینولایتیکوس بود، براساس نتایج دامنه میزبانی، فاژ Vibrio alginolyticus strain Samira Phage قادر به تشکیل پلاگ بود.نتیجه گیری: باکتریوفاژ Vibrio alginolyticus strain Samira Phage جداسازی شده اثر باکتری کشی با دامنه ای وسیع در برابر باکتری های ویبریوی شناسایی شده از سواحل بوشهر داشت که نشان می دهد این باکتریوفاژ می تواند کاندیدی مناسبی برای استفاده در فاژ درمانی بر علیه بیماری ویبریوز باشد.
The aim of this study was to screen Vibrio species off the coast of Bushehr province and control this shrimp pathogen using phage.For this study, water was sampled in 1398 from areas close to the coast of Bushehr province. Bacteria that cause Vibriosis were determined using biochemical tests and molecular sequencing. Bacterial selection was performed according to the results of antibiogram profiles of dominant bacteria and pathogens. TCBS was used to determine the phage titer of the culture medium and the plugs were counted 24 hours after incubation at 30 ° C.5 bacteria (Vibrio alginolyticus strain SeqID, Vibrio xuii strain Z-03Phage, Vibrio alginolyticus SZ / 10 gene, Vibrio rotiferianus strain SRPR-Phage, Vibrio harveyi strain Z05 were isolated from Bushehr wastewater contaminated waters). Vibrio alginolyticus strain SeqID was selected as the dominant bacterial species according to the results of antibiogram profile determination, growth halo diameter and bacterial biochemical test of the studied species. Based on the results of 16srDNA molecular analysis, the isolated strain of the bacterium was 100% similar to Vibrio alginolyticus. Based on the results of the host domain, Vibrio alginolyticus strain Samira Phage was able to form plugs.The isolated bacteriophage Vibrio alginolyticus strain Samira Phage had a bactericidal effect with a wide range against Vibrio bacteria identified from the shores of Bushehr, which indicates that this bacteriophage can be a good candidate for use in phage therapy Be against viberosis.Keywords: Isolation and screening, Bacteriophage, Vibriosis, Western white shrimp
1.Amatul-Samahah, MA., Omar, W., Ikhsan, NFM., Azmai, MNA., Zamri-Saad, M., Ina-Salwany, MY. (2020). Vaccination trials against vibriosis in shrimp: A review. Aquaculture Reports, 18 (3);16-22.1.
2.Bell, JD., Munro, JL., Nash, WJ., Rothlisberg, PC., Loneragan, NR., Ward, RD. (2005). Stock enhancement initiatives. restocking and stock enhancement of marine invertebrate fisheries. Academic Press, 22(5); 143–196.
3.Benyajirapatch, D., Rengpipat, S. (2014). Isolation and characterization of bacteriophages specific to Vibrio cholera. Aquaculture Reports, 13(9);19-26.
4.Choudhury, TG,, Maiti, B., Venugopal, MN., Karunasagar, I. (2012). Effect of total dissolved solids and temperature on bacteriophage therapy against Luminous vibriosis in Shrimp. Applied Microbiology and Biotechnology, 54(9);12-16.
5.Culot. A., Grosset, N., Gautier, M. (2019). Overcoming the challenges of phage therapy for industrial aquaculture: A review. Aquaculture, 513; 734423.
6.Ding, T., Sun, H., Pan, Q., Zhao, F., Zhang, Z., Ren, H, (2020). Isolation and characterization of Vibrio parahaemolyticus bacterio phage vB_VpaS_PG07. Virus Research, 286; 198080.
7.Hou, D., Huang, Z., Zeng, S., Liu, J., Wei, D., Deng, X. (2018). Intestinal bacterial signatures of white feces syndrome in shrimp. Applied Microbiology and Biotechnology, 102; 3701–3709.
8.Hu, F., Guo, Y., Yang, Y., Zheng, Y., Wu, S., Jiang, X. (2019). Resistance reported from china antimicrobial surveillance network (CHINET) in 2018. European Journal of Clinical Microbiology & Infectious Diseases, 38; 2275–2281.
9.Ibrahim, WNW., Leong, LK., Razzak, LA., Musa, N., Danish-Daniel, M., Zainathan, SC. (2021). Virulence properties and pathogenicity of multidrug-resistant Vibrio harveyi associated with luminescent vibriosis in pacific white shrimp, Penaeus vannamei. Journal of Invertebrate Pathology, 107-94.
10.Janda, JM., Newton, AE., Bopp, CA. (2015). Vibriosis. clinics in laboratory medicine. Journal of Invertebrate Pathology, 35; 273–288.
11.Jun JW, Kim HJ, Yun SK, Chai JY, Park SC (2014) Eating oysters without risk of vibriosis: application of a bacteriophage against Vibrio parahaemolyticus in oysters. International journal of food microbiology 188:31–35
12.Kais, SM. (2019). Climate change: vulnerability and resilience in commercial shrimp aquaculture in bangladesh. oceanography and coastal informatics: breakthroughs in research and practice. IGI Global, pp 152–177.
13.Kalatzis, PG., Bastías, R., Kokkari, C., Katharios, P. (2016). Isolation and characterization of two lytic bacteriophages, φSt2 and φGrn1; phage therapy application for biological control of Vibrio alginolyticus in aquaculture live feeds. PloS one, 11; e0151101.
14.Kalatzis, PG., Bastías, R., Kokkari, C., Katharios, P. (2016). Isolation and characterization of two lytic bacteriophages, φSt2 and φGrn1; phage therapy application for biological control of Vibrio alginolyticus in aquaculture live feeds. PloS one, 11; e0151101.
15.Karunasagar, I., Shivu, M., Girisha, S., Krohne, G., Karunasagar, I. (2007). Biocontrol of pathogens in shrimp hatcheries using bacteriophages. Aquaculture, 268; 288–292.
16.Letchumanan, V., Chan, K-G., Pusparajah, P., Saokaew, S., Duangjai, A., Goh, B-H. (2016). Insights into bacteriophage application in controlling Vibrio species. Frontiers in Microbiology, 7;11-14.
17.Lomelí-Ortega, CO., Martínez-Díaz, SF. (2014). Phage therapy against Vibrio parahaemolyticus infection in the whiteleg shrimp(Litopenaeus vannamei) larvae. Aquaculture, 434; 208–211.
18.Luna, L., Hernández, D., Silva, HV., Cobos, MA., González, SS., Cortez, C. (2019). Isolation, biochemical characterization, and phylogeny of a cellulose-degrading ruminal bacterium. Revista Colombiana de Ciencias Pecuarias, 32;117–125.
19.Luo, P., He, X., Liu, Q., Hu, C. (2015). Developing universal genetic tools for rapid and efficient deletion mutation in Vibrio species based on suicide T-vectors carrying a novel counterselectable marker, vmi480. PLoS One, 10; e0144465.
20.Martínez-Díaz, SF., Hipólito-Morales, A. (2013). Efficacy of phage therapy to prevent mortality during the vibriosis of brine shrimp. Aquaculture, 400–401; 120–124.
21.Mateus, L., Costa, L., Silva, YJ., Pereira, C., Cunha, A., Almeida, A. (2014). Efficiency of phage cocktails in the inactivation of Vibrio in aquaculture. Aquaculture, 424–425; 167–173.
22.Matsuzaki, S., Tanaka, S., Koga, T., Kawata, T. (1992). A broad‐host‐range vibriophage, KVP40, isolated from sea water. Microbiology and Immunology, 36; 93–97.
23.Mirbakhsh, M., Zorriehzahra, S., Yeganeh, V., Ghaednia, B., Dashtiannasab, A., Mohammadi Baghmollai, E. (2016). Molecular identification of bacterial and fungal pathogens in certain disease-free shrimp. Journal of Sea Research, 76 (8);231-240.
24.Onarinde, BA., Dixon, RA. (2018). Prospects for biocontrol of Vibrio parahaemolyticus Contamination in Blue Mussels(Mytilus edulus) a year-long study. Frontiers in Microbiology 9:1043.
25.Phumkhachorn, P., Rattanachaikunsopon, P. (2010). Isolation and partial characterization of a bacteriophage infecting the shrimp pathogen Vibrio harveyi. African Journal of Microbiology Research, 4; 1794–1800.
26.Plaza, N., Castillo, D., Pérez-Reytor, D., Higuera, G., García, K., Bastías, R. (2018). Bacteriophages in the control of pathogenic vibrios. Electronic Journal of Biotechnology 31; 24–33.
27.Ramasamy, P. (2019). Phage therapy for control of bacterial diseases. Crustacea, P.89-90.
28.Rasmussen, BB., Kalatzis, PG., Middelboe, M., Gram, L. (2019). Combining probiotic Phaeobacter inhibens DSM17395 and broad-host-range vibriophage KVP40 against fish pathogenic vibrios. Aquaculture, 513;734415.
29.Rezaie, N., Pourshafie, M. (2018). Increased resistance to tetracycline and erythromycin in Vibrio cholerae. clinical isolates isolated from patients with cholera disease during 2012-2013 outbreaks in IR Iran. Infection Epidemiology and Microbiology, 4; 93–98.
30.Ringø, E. (2020). Probiotics in shell fish aquaculture. Aquaculture and Fisheries, 5;1–27.
31.Rowley, AF., Pope, EC. (2012). Vaccines and crustacean aquaculture. A mechanistic exploration. Aquaculture, 334–337; 1–11.
32.Sasikala, D., Srinivasan, P. (2016). Characterization of potential lytic bacteriophage against Vibrio alginolyticus and its therapeutic implications on biofilm dispersal. Microbial Pathogenesis, 101; 24–35.
33.Shinn, A., Pratoomyot, J., Griffiths, D., Trong, T., Vu, NT., Jiravanichpaisal, P. (2018). Asian shrimp production and the economic costs of disease. Asian Fish Sci S., 31; 29–58.
34.Silva, YJ., Costa, L., Pereira, C., Mateus, C., Cunha, A., Calado, R. (2014). Phage therapy as an approach to prevent Vibrio anguillarum infections in fish larvae production. PLoS One, 9; e114197
35.Stalin, N., Srinivasan, P. (2017). Efficacy of potential phage cocktails against Vibrio harveyi and closely related Vibrio species isolated from shrimp aquaculture environment in the south east coast of India. Veterinary Microbiology, 207;83-96.
36.Sweet, MJ., Bateman, KS. (2015). Diseases in marine invertebrates associated with mariculture and commercial fisheries. Journal of Sea Research, 104; 16–32.
37.Sweet, MJ., Bateman, KS. (2016). Reprint of ‘diseases in marine invertebrates associated with mariculture and commercial fisheries. Journal of Sea Research, 113; 28–44.
38.Valente, C de S., Wan, AHL. (2021). Vibrio and major commercially important vibriosis diseases in decapod crustaceans. Journal of Invertebrate Pathology, 181; 107527
39.Vinod, MG., Shivu, MM., Umesha, K., Rajeeva, B., Krohne, G., Karunasagar, I. (2006). Isolation of Vibrio harveyi bacteriophage with a potential for biocontrol of luminous vibriosis in hatchery environments. Aquaculture, 255; 117–124.
40.Wang, D., Mbewe, N., Bels, LD., Couck, L., Stappen, GV., Broeck, WV. (2021). Pathogenesis of experimental vibriosis in blue mussel (Mytilus edulis) larvae based on accurate positioning of GFP-tagged Vibrio strains and histopathological and ultrastructural changes of the host. Aquaculture, 535; 736347.
41.Wang, A., Ran, C., Wang, Y., Zhang, Z., Ding, Q., Yang, Y. (2019). Use of probiotics in aquaculture of China. A review of the past decade. Fish & Shellfish Immunology, 86;734–755.
42.Xie, J., Bu, L., Jin, S., Wang, X., Zhao, Q., Zhou, S. (2020). Out break of vibriosis caused by Vibrio harveyi and Vibrio alginolyticus in farmed seahorse Hippocampus kuda in China. Aquaculture, 523; 735168.
43.Yang, M., Liang, Y., Huang, S., Zhang, J., Wang, J., Chen, H. (2020). Isolation and characterization of the novel phages vB_VpS_BA3 and vB_VpS_CA8 for lysing Vibrio parahaemolyticus. Frontiers in Microbiology, 11; 259.
44.Yuksel, SA., Thompson, KD., Ellis, AE., Adams, A. (2001). Purification of Piscirickettsia Salmonis And Associated Phage Particles. Diseases of Aquatic Organisms, 44; 231–235-309.
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