The Isolation and Examination of the Host Specificity of Local Bacteriophages for Use Against Brucella abortus

Year 2023, Volume: 12 Issue: 1, 80 - 85, 22.06.2023

Songül Ötkün Sevil Erdenlig Gürbilek Ahmet Murat Saytekin

https://doi.org/10.31196/huvfd.1293366

Abstract

Bacteriophages are viruses that infect bacteria. Although their effects on bacteria have been known for many years, the discovery of antibiotics has limited the widespread use of bacteriophages against bacterial infections. However, it is now essential to reconsider using phage therapies due to antimicrobial-resistant bacteria and uncontrolled bacterial zoonotic diseases becoming a global problem. This study aimed to isolate lytic phages against B. abortus, the most common agent that causes bovine brucellosis, which has become a worldwide animal and public health problem. This performed isolation is from cattle farms known to be previously or currently infected, and the study also evaluates the lytic effects of these phages on B. abortus, B. melitensis, B. suis biotypes, B. ovis and B. neotomae and B. abortus field isolates in our culture collection. In this process, seven local brucella-specific phages were identified by evaluating 112 samples via general phage isolation and purification. The lyticity of the isolated bacteriophages were analyzed with international reference: B. abortus (bv 1, 2, 3, 4, 5, 6, 9), B. melitensis (bv 1, 2, 3), B. suis (bv 1, 2, 3, 4, 5) biovars, B. ovis, B. neotomae and B. abortus field strains (n:20). It was found that 85% of B. abortus strains produced a lysis pattern like Tbilisi Φ through local phages. In terms of the lysis results, three different B. abortus specific phages were isolated (98 Φ, 104 Φ, and (P35, P70, P94/1, P94/2, P94/3) phages). It is thought that the applying cocktails prepared from these phages to fight against brucellosis will significantly contribute to controlling the disease. Since 15% of the field isolates were found to be rough strains, it is recommended that R/C Φ are included in the prepared cocktails.

Keywords

Bacteriophage, phage therapy, B. abortus, brucellosis

Brucella abortus’a Karşı Yerel Bakteriyofaj İzolasyonu ve Konakçı Spesifitesinin Değerlendirilmesi

Abstract

Bakteriyofajlar bakterileri enfekte eden viruslardır. Bakteriler üzerindeki etkileri uzun yıllardır bilinmekle birlikte bakteriyel enfeksiyonlara karşı yaygın olarak kullanımı antibiyotiklerin keşfi nedeniyle arka planda kalmıştır. Küresel bir sorun haline gelen antimikrobiyal dirençli bakteriler ve kontrol altına alınamayan bakteriyel zoonotik hastalıklar nedeniyle günümüzde faj terapileri bir tercihten ziyade zorunluluk haline gelmeye başlamıştır. Bu çalışmada, ülkemizde ve dünyanın birçok ülkesinde önemli bir hayvan ve halk sağlığı sorunu olan sığır brusellosisinin en yaygın etkeni olan B. abortus’a karşı daha önce ya da halen infekte olduğu bilinen sığır çiftliklerinden litik faj izolasyonunun yapılması ve bu fajların B. abortus, B. melitensis B. suis biyotiplerinde, B. ovis ve B. neotomae’de ve kültür koleksiyonumuzda bulunan B. abortus saha izolatlarında litik etkilerinin değerlendirilmesi amaçlanmıştır. Genel faj izolasyon ve saflaştırma yöntemleri uygulanarak 112 örnekten yedi adet yerel Brucella spp. spesifik faj tespit edildi. İzole edilen bakteriyofajlar, uluslararası referans B. abortus (bv 1,2,3,4,5,6,9), B. melitensis (bv 1,2,3), B. suis (bv 1,2,3,4,5) biyovarları, B. ovis, B. neotomae ve B. abortus saha suşları (n:20) ile litik etkinlikleri yönünden analiz edildi. B. abortus suşlarının %85’i yerel fajlarla Tbilisi Φ’na benzer lizis modeli meydana getirdi. Çalışma sonunda şekillenen lizis tablosu dikkate alındığında, 3 farklı B. abortus spesifik faj izolasyonunun yapıldığı anlaşıldı [98 Φ, 104 Φ, ve (P35, P70, P94/1, P94/2, P94/3) fajları]. Bu fajlardan hazırlanan faj karışımlarının brusellosis ile mücadelede uygulanmasının hastalığın kontrolüne önemli katkılar sağlayacağı düşünülmektedir. Çalışmada saha izolatlarının %15’i rough suşlar olarak tespit edildiğinden hazırlanan faj kokteyllerin içine R/C Φ’nın katılmasının da yerinde bir karar olacağı kanısına varıldı.

Keywords

Bakteriyofaj, faj terapisi, B. abortus, brusellosis

References

• Alton GG, Jones LM, Angus RD, Verger JM, 1988: Techniques For The Brucellosis Laboratory. Paris: INRA.
• Aslam S, Schooley RT, 2019: What’s old is new again: bacteriophage therapy in the 21st century. Antimicrob Agents Chemother, 64 (1), e01987-19.
• Campbell A, 2003: The future of bacteriophage biology. Nat Rev Genet, 4 (6), 471-7.
• Chachra D, Kaur H, Chandra M, Saxena HM, 2012: Isolation, electron microscopy and physicochemical characterization of a brucella phage against Brucella abortus vaccine strain S19. Internet J Microbiol, 10 (2), 1-7.
• Flores V, López-Merino A, Mendoza-Hernandez G, Guarneros G, 2012: Comparative genomic analysis of two brucella phages of distant origins. Genomics, 99 (4), 233-240.
• García R, Latz S, Romero J, Higuera G, García K, Bastías R, 2019: Bacteriophage production models: an overview. Front Microbiol, 10, 1187.
• Gupta V, Saxena HM 2017: Isolation and characterization of BpL1, a broad acting lytic bacteriophage against Brucella. Int J Curr Microbiol Appl Sci, 6 (11), 2486-2496.
• Erdenliğ-Gürbilek S, Arserim NB, Tel OY, 2022: Determination of serogroup and lytic activities of bacteriophages isolated from phage plaques in Staphylococcus aureus cultures identified from sheep milk with mastitis. Indian J of Anim Res, 1, 4.
• Hammerl JA, Göllner C, Jäckel C, Scholz HC, Nöckler K, Reetz, J, Dahouk, Hertwig S, 2017: Genetic diversity of Brucella reference and non-reference phages and its impact on Brucella-typing. Front Microbiol, 8, 408.
• Issabekov SS, Syrym NS, Sambetbayev AA, Alikhanov KD, Yespembetov BA, 2022: Prospects of bacteriophage collections in disinfectant applications. Veterinary World, 15 (1), 220-231.
• Jurač K, Nabergoj D, Podgornik A, 2019: Bacteriophage production processes. Appl Microbiol Biotechnol, 103 (2), 685-694.
• Khurana SK, Sehrawat A, Tiwari R, Prasad M, Gulati B, Shabbir MZ, Chhabra R, Karthik K, Patel SK, Pathak M, Yatoo MI, Gupta K, Sah R, Chaicumpa, W 2021: Bovine brucellosis–a comprehensive review. Veterinary Quarterly, 41 (1), 61-88.
• Li XM, Kang YX, Lin L, Jia EH, Piao DR, Jiang H, Zhang CC, He J, Chang YF, Guo XK, Zhu, Y, 2019: Genomic characterization provides new insights for detailed phage-resistant mechanism for Brucella abortus. Front Microbiol, 10, 917.
• Ling H, Lou X, Luo Q, He Z, Sun M, Sun J, 2022: Recent advances in bacteriophage-based therapeutics: Insight into the post-antibiotic era. Acta Pharm Sin B, 12 (12), 4348-4364.
• Mohan A, Saxena HM, 2020: Effect of phage targeting therapy of brucellosis on host antibody response in cattle. Phage (New Rochelle), 1 (4), 223-229.
• Morris JA, Corbel MJ, Phillip JIH, 1973: Characterization of three phages lytic for Brucella species. J of Gen Virol, 20 (1), 63-73.
• Pappas G, Papadimitriou P, Akritidis N, Christou L, Tsianos EV, 2006: The new global map of human brucellosis. Lancet Infec Dis, 6 (2), 91-99.
• Phongtang W, Choi GP, Chukeatirote E, Ahn J, 2019: Bacteriophage control of Salmonella Typhimurium in milk. Food Sci Biotechnol, 28, 297-301.
• Prajapati A, Ramchandran D, Verma H, Abbas M, Rawat M, 2014: Therapeutic efficacy of Brucella phage against Brucella abortus in mice model. Vet World, 7 (1), 34-37.
• Projahn M, Hammerl JA, Dieckmann R, Dahouk SA, 2020: A proof of principle for the detection of viable Brucella spp. in raw milk by qPCR targeting bacteriophages. Microorganisms, 8 (9), 1326.
• Sambrook J, Fritsch EF, Maniatis T, 1989: Molecular cloning: a laboratory manual, 2nd ed., Cold Spring Harbor Laboratory press, ABD.
• Saxena HM, 2021: Bacteriophage and its potential for therapeutic use in brucellosis among cattles. Research & Reviews: J Vet Sci Technol, 10 (2), 9–17.
• Shaheen AY, Sheikh AA, Rabbani M, Shehzad W, Abbas Z, Maqbool M, 2021: Isolation, propagation and biocontrol activity of indigenous bacteriophages against Brucella abortus. Intl J Agric Biol, 25, 1066‒1074.
• Tevdoradze E, Farlow J, Kotorashvili A, Skhirtladze N, Antadze I, Gunia S, Balarjishvili, Kvachadze L, Kutateladze M, 2015: Whole genome sequence comparison of ten diagnostic Brucella phages propagated on two Brucella abortus hosts. Virol J, 12 (1), 1-11.
• World Organisation for Animal Health, 2022: Chapter 3. 1. 4. Brucellosis (Infection with B. abortus, B. melitensis and B. suis). In Manual of Diagnostic Tests and Vaccines for Terrestrial Animals, Paris.