Sığır Abort Materyallerinden Salmonella Dublin İdentifikasyonu veFilogenetik Pozisyonlandırılması

Bu çalışmanın amacı Türkiye’nin dört farklı şehrinden (Ağrı, Erzincan, Erzurum ve Kars) alınan 367 adet sığır aborte fötus örneklerine ait karaciğer ve akciğer dokuları ile abomazum içeriklerinde Salmonella Dublin’i izole etmek ve moleküler yöntemlerle karakterizasyonunu sağlamaktır. Doku ve içeriklerin uygun inkübasyonu sonrasında Salmonella spp. identifikasyonu yapıldı. DNA ekstraksiyonu için Salmonella spp. şüpheli kolonilere kuru ısı bloğu üzerinde kaynatma metodu, Salmonella spp.’ların tür düzeyinde identifikasyonu ve S. Dublin teşhisi için ise multipleks PCR metodu kullanıldı. Multipleks PCR analizi sonucunda S. Dublin’e özgü bant profili gösteren bir izolatın 16S rDNA dizi analizi ile sekanslaması ve filogenetik analizi gerçekleştirildi. S. Dublin identifikasyonu ise BLAST analizine göre ve NCBI GenBank’tan elde edilen benzerlik skorlarına göre yapıldı. Sığır atık fötal örneklerinde S. Dublin’in moleküler prevalansı %0.82 olarak saptandı. İzolatın 16S rRNA sekans bulguları birçok S. Dublin suşu ile benzer olarak bulundu. İzolatın 16S rRNA gen bölgesinin sekansı Genbankasına MZ452230 erişim numarası ve Erzurum VKEM suş ismi ile kaydedilmiştir. Sonuç olarak sığır atık fötusundan S. Dublin tanımlanmış ve 16S rDNA gen bölgesinin dizi analizi ile de S. Dublin’e filogenetik pozisyon kazandırılmıştır.

Identification and Phylogenetic Positioning of Salmonella Dublin from Aborted Cattle Materials

This study was aimed to isolate, and characterize the Salmonella Dublin in the liver, lung tissues and the abomasum contents of 367 aborted cattle fetal samples obtained from four different cities (Ağrı, Erzincan, Erzurum and Kars) of Turkey by using molecular methods. After proper incubation of the tissues and contents, Salmonella spp. identification was performed. Colonies with suspicion of Salmonella were boiled on a dry heat block for DNA extraction, while multiplex PCR was used for species identification of Salmonella spp. and identification of S. Dublin. The multiplex PCR, 16S rDNA sequence, and phylogenetic analysis were performed on an isolate with a band profile specific to S. Dublin. The S. Dublin identification was performed according to BLAST analysis and similarity scores obtained from NCBI GenBank. Molecular prevalence of S. Dublin in the aborted fetal samples of cattle was 0.82%. The 16S rRNA sequence results of the isolate was found to be similar to many S. Dublin strains. The partial sequence of the 16S rRNA gene region of the isolate was recorded in the GenBank database with the name strain Erzurum VCRI and access number MZ452230. In conclusion, S. Dublin was identified in the aborted cattle fetuses and phylogenetic position was determined for S. Dublin using the sequence analysis of 16S rDNA gene region.

PDF

___

1. Holschbach CL, Peek SF: Salmonella in dairy cattle. Vet Clin North Am Food Anim Pract, 34 (1): 133-154, 2018. DOI: 10.1016/j.cvfa.2017.10.005
2. Nielsen LR: Review of pathogenesis and diagnostic methods of immediate relevance for epidemiology and control of Salmonella Dublin in cattle. Vet Microbiol, 162 (1): 1-9, 2013. DOI: 10.1016/j.vetmic.2012.08.003
3. Hezil D, Zaidi S, Benseghir H, Zineddine R, Benamrouche N, Ghalmi F: Salmonella Dublin associated with abortion in dairy cattle in Algiers and comparison of different diagnostic methods. Afr J Clin Exp Microbiol, 22 (2): 211-222, 2021. DOI: 10.4314/ajcem.v22i2.14
4. Peek SF, Mcguirk SM, Sweeney RW, Cummings KJ: Infectious diseases of the gastrointestinal tract. Rebhun’s Diseases of Dairy Cattle. 3rd ed., 249-356, 2018. DOI: 10.1016/B978-0-323-39055-2.00006-1
5.Hinton M: The diagnosis of salmonella abortion in cattle with particular reference to Salmonella Dublin. A review. Epidemiol Infect, 79 (1): 25-38, 1977. DOI: 10.1017/s0022172400052815
6. Sodoma E, Mitterhuemer S, Krassnig G, Stellnberger K, Reisp K, Schmoll F, Dünser M: Infektiös bedingte Aborte beim Rind-eigene Erfahrungen und Untersuchungen aus dem Jahr 2018 (Januar-September). Tierärztl Prax Ausg G: Grosstiere Nutztiere, 47, 143-150, 2019. DOI: 10.1055/ a-0896-0945
7. Anderson ML: Infectious causes of bovine abortion during mid-to late-gestation. Theriogenology, 68 (3): 474-486, 2007. DOI: 10.1016/j. theriogenology.2007.04.001
8. Carrique-Mas J, Willmington J, Papadopoulou C, Watson E, Davies R: Salmonella infection in cattle in Great Britain, 2003 to 2008. Vet Rec, 167 (15): 560-565, 2010. DOI: 10.1136/vr.c4943
9. Noakes DE, Parkinson TJ, England GC: Subfertility and infertility. In, Veterinary Reproduction and Obstetrics 9th ed., 393-646, Saunders Elseiver, England, 2018.
10. OIE Manual of Standards for Diagnostic Tests and Vaccines for Terrestrial Animals: Ch. 3.9.8. Salmonellosis (NB: Version adopted in May 2016), 2018.
11. Babacan O, Karadeniz H: Çiğ tavuk etlerinden izole edilen Salmonella spp. suşlarının antibiyotik duyarlılıklarının araştırılması. Vet Hekim Der Derg, 90 (2): 105-114, 2019. DOI: 10.33188/vetheder.497569
12. Zhai L, Kong X, Lu Z, Lv F, Zhang C, Bie X: Detection of Salmonella enterica serovar Dublin by polymerase chain reaction in multiplex format. J Microbiol Methods, 100, 52-57, 2014. DOI: 10.1016/j.mimet.2014.02.014
13. Fredriksson NJ, Hermansson M, Wilén BM: The choice of PCR primers has great impact on assessments of bacterial community diversity and dynamics in a wastewater treatment plant. PloS One, 8 (10): e76431, 2013. DOI: 10.1371/journal.pone.0076431
14. Petti CA, Brandt ME, Church DL, Emler S, Simmon K, Zelazny AM: Interpretive criteria for identification of bacteria and fungi by DNA target sequencing. In, CLSI guideline MM18, 2nd ed., 7-117, Clinical and Laboratory Standards Institute, Philadephia, 2018.
15. Nishimaki T, Sato K: An Extension of the Kimura two-parameter model to the natural evolutionary process. J Mol Evol, 87 (1): 60-67, 2019. DOI: 10.1007/s00239-018-9885-1
16. Barkallah M, Gharbi Y, Hassena AB, Slima AB, Mallek Z, Gautier M, Greub G, Gdoura R, Fendri I: Survey of infectious etiologies of bovine abortion during mid-to late gestation in dairy herds. PLoS One, 9 (3): e91549, 2014. DOI: 10.1371/journal.pone.0091549
17. Anderson ML, Blanchard PC, Barr BC, Hoffman RL: A survey of causes of bovine abortion occurring in the San Joaquin Valley, California. J Vet Diagn Invest, 2 (4): 283-287, 1990. DOI: 10.1177/104063879000200405
18. Halimi HA, Seifi HA, Rad M: Bovine salmonellosis in Northeast of Iran: Frequency, genetic fingerprinting and antimicrobial resistance patterns of Salmonella spp. Asian Pac J Trop Biomed, 4 (1): 1-7, 2014. DOI: 10.1016/S2221-1691(14)60199-4
19. Kirkbride CA: Bacterial agents detected in a 10-year study of bovine abortions and stillbirths. J Vet Diagn Invest, 5 (1): 64-68, 1993. DOI: 10.1177/104063879300500114
20.Derdour SY, Hafsi F, Azzag N, Tennah S, Laamari A, China B, Ghalmi F: Prevalence of the main infectious causes of abortion in dairy cattle in Algeria. J Vet Res, 61 (3): 337-343, 2017. DOI: 10.1515/jvetres-2017-0044
21. Reichel MP, Wahl LC, Hill FI: Review of diagnostic procedures and approaches to infectious causes of reproductive failures of cattle in Australia and New Zealand. Front Vet Sci, 5:222, 2018. DOI: 10.3389/ fvets.2018.00222
22. Regional Veterinary Laboratories Report - November 2014: Vet Ireland J, 5, 79-84, 2015.
23. Geisbauer E, Stellnberger K, Krassnig G, & Dünser M: Salmonella dublin outbreak in cattle. Animal Hygiene and Sustainable Livestock Production. Proceedings of the XVth International Congress of the International Society for Animal Hygiene, Vienna, Austria, 575, 3-7 July2011.
24. Jerrett IV, McOrist S, Waddington J, Browning JW, Malecki JC, McCausland IP: Diagnostic studies of the fetus, placenta and maternal blood from 265 bovine abortions. Cornell Vet, 74 (1): 8-20, 1984.
25. Karatas Yeni D, Akca D: Evaluation of the analytical efficiency of RealTime PCR in the diagnosis of Brucellosis in cattle and sheep. Kafkas Univ Vet Fak Derg, 27 (4): 503-509, 2021. DOI: 10.9775/kvfd.2021.25776
26.Drancourt M, Bollet C, Carlioz A, Martelin R, Gayral JP, Raoult D: 16S ribosomal DNA sequence analysis of a large collection of environmental and clinical unidentifiable bacterial isolates. J Clin Microbiol, 38 (10): 3623- 3630, 2000. DOI: 10.1128/JCM.38.10.3623-3630.2000