Evaluation of pathogenic serovars of Leptospira interrogans in dairy cattle herds of Shahrekord by PCR
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Abstract
Background and objectives: Leptospirosis is an important zoonotic disease caused by Leptospira interrogans. Leptospirosis leads to economical losses in dairy farm industry. The objective of this study was to evaluate the pathogenic serovars of Leptospira interrogans in dairy cattle herds of Shahrekord by PCR.
Materials and Methods: Two hundred samples (100 urine and 100 blood) were collected from 100 cows randomly and delivered to the laboratory. Samples were stored at −20 °C. DNA was extracted and purified from the plasma and urine samples and concentrated on diatoms in the presence of guanidine thiocyanate (GuSCN). PCR products were detected and identified as Leptospira by ilumination of the expected size of DNA bands after staining of the agarose gel with ethidium bromide gels. PCR products were purified and sequenced.
Results: The results showed that 28% of urine samples and 23% of plasma samples were contaminated. The major serotypes were Icterohaemorrhagiae (50%) and Pomona (37.5%). The urine samples of 17 cows were positive for Leptospira without positive plasma samples. This indicated that these cows are reservoirs in dairy herds of Shahrekord and dangerous for human health. The plasma samples of twelve cows were positive for Leptospira without positive urine samples.
Conclusions: Leptospira serotypes can be maintained in relatively dry regions and must be considered when dealing with leptospirosis in dairy farms of Shahrekord and human health.
Radostits OM, Gay CC, Hinchcliff KW, Constable PD. (2007) Veterinary Medicine: a textbook of the diseases of cattle, horses, sheep, pigs and goats.10th ed. London, New York.
Ahmed N, Devi SM, Valverde M, Vijayachari P, Machangu RS, Ellis WA, et al. Multilocus sequence typing method for identification and genotypic classification of pathogenic Leptospira species. Ann Clin Microbiol Antimicrob 2006; 5: 1-10.
Palaniappan RUM, Chang YF, Chang CF, Pan MJ, Yang CW, Harpending P, et al. Evaluation of lig-based conventional and real time PCR for the detection of pathogenic leptospires. Mol Cell Probes 2005; 19: 111-117.
Ellis WA. Bovine leptospirosis in the tropics: prevalence,pathogenesis and control. Prev Vet Med 1984; 2 : 411-421.
Levett PN, Branch SL, Whittington CU, Edwards CN, Paxton H. Two methods for rapid serological diagnosis of acute leptospirosis. Clin Vaccine Immuno 2001; 8:349-351.
Honarmand HR, Mansour Ghanaei F, Heydarzadeh A. Isolation and Serotyping of endemic leptospires of eastern part of flat area of Guilan province, Iran. J Gorgan Uni Medl Sci 2009; 11: 53-59.
Levett PN, Morey RE, Galloway RL, Turner DE, Steigerwalt AG, Mayer LW. Detection of pathogenic leptospires by real-time quantitative PCR. J Med Microbiol 2005; 54: 45-49.
Bolin CA, Alt DP. Use of a monovalent leptospiral vaccine to prevent renal colonization and urinary shedding in cattle exposed to Leptospira borgpetersenii serovar hardjo. Am J Vet Res 2001; 62: 995-1000.
Ebrahimi A, Nasr Z, Kojouri GA. Seroinvestigation of bovine leptospirosis in Shahrekord district, central Iran. Iran J Vet Res 2004; 5: 110-113.
Thiermann AB. Leptospirosis: current developments and trends. J Am Vet Med Assoc 1984; 184: 722-725.
Mendoza L, Prescott JF. Serodiagnosis of leptospirosis in pigs using an axial filament enzyme-linked immunosorbent assay. Vet Microbiol 1992; 31: 55-70.
Surujballi O, Henning D, Marenger R, Howlett C. Development of a monoclonal antibody-based competitive enzyme-linked immunosorbent assay for the detection of Leptospira borgpetersenii serovar hardjo type hardjobovis antibodies in bovine sera. Can J Vet Res 1997; 61: 267-274.
Jouglard SDD, Simionatto S, Seixas FK, Nassi FL, Dellagostin OA. Nested polymerase chain reaction for detection of pathogenic leptospires. Can J Microbiol 2006; 52: 747-752.
Terpstra WJ, Schoone GJ, Ter Schegget J. Detection of leptospiral DNA by nucleic acid hybridisation with 32P- and biotin-labelled probes. Jo Medl Microbiol 1986; 22:23-28.
Terpstra WJ, Schoone GJ, Ligthart GST, Schegget J. Detection of Leptospira interrogans in clinical specimens by in situ hybridization using biotin-labelled DNA probes. Microbiol 1987; 133: 911-914.
LeFebvre RB, Thiermann AB, Foley J. Genetic and antigenic differences of serologically indistinguishable leptospires of serovar hardjo. J Clin Microbiol 1987; 25:2094-2097.
Alt DP, Zuerner RL, Bolin CA. Evaluation of antibiotics for treatment of cattle infected with Leptospira borgpetersenii serovar hardjo. J Am Vet Med Assoc 2001; 219: 636-639.
Gerritsen MJ, Olyhoek T, Smits MA, Bokhout BA.Sample preparation method for polymerase chain reaction-based semiquantitative detection of Leptospira interrogans serovar hardjo subtype hardjobovis in bovine urine. J Clin Microbiol 1991; 29: 2805-2808.
Taylor MJ, Ellis WA, Montgomery JM, Yan KT, McDowell SWJ, Mackie DP. Magnetic immuno capture PCR assay (MIPA): detection of Leptospira borgpetersenii serovar hardjo. Vet Microbiol 1997; 56 (1-2): 135-145.
Wagenaar J, Zuerner RL, Alt D, Bolin CA. Comparison of polymerase chain reaction assays with bacteriologic culture, immunofluorescence, and nucleic acid hybridization for detection of Leptospira borgpetersenii serovar hardjo in urine of cattle. Am J Vet Res 2000;61:316-320.
Wagenaar JA, Segers RPAM, Van der Zeijst BAM.Rapid and specific detection of pathogenicLeptospira species by amplification of ribosomal sequences. Mol Biotechnol 1994; 2: 1-14.
Talpada MD, Garvey N, Sprowls R, Eugster AK, Vinetz JM. Prevalence of leptospiral infection in Texas cattle: implications for transmission to humans. Vector Borne Zoonotic Dis 2003; 3: 141-147.
Nassi F, Seixas FK, Jouglard SDD, Simionatto S, Silva EF, Seyffert N, et al. Leptospirosis diagnosis using Nested-PCR. Braz J Microbiol 2003; 34: 90-92.
Brown PD, Gravekamp C, Carrington DG, Van de Kemp H, Hartskeerl RA, Edwards CN, et al. Evaluation of the polymerase chain reaction for early diagnosis of leptospirosis. J Med Microbiol 1995; 43: 110-114.
Pavan ME, Cair F, Brihuega B, Samartino L. Multiple- locus variable-number tandem repeat analysis (MLVA) of Leptospira interrogans serovar Pomona from Argentina reveals four new genotypes. Comp Immunol Microbiol Infect Dis 2008; 31: 37-45.
Jolley KA, Feil EJ, Chan MS, Maiden MCJ. Sequence type analysis and recombinational tests (START). Bioinformatics 2001; 17: 1230-1231.
Van Eys GJ, Gravekamp C, Gerritsen MJ, Quint W, Cornelissen MT, ScHegget JT, et al. Detection of leptospires in urine by polymerase chain reaction. J Clin Microbiol 1989; 27 : 2258-2262.
Kiamarzi G. seroepidemiology of leptospirosis in cowa in Shahrekord. Thesis of DVM, Azad University, Shahrekord 1998.
Rodrigues CG, Müller EE, Freitas JC. Bovine leptospirosis, serology at dairy farms in Londrina region, Paran State, Brazil. Cienc Rural 1999; 29: 309-314.
Hajj Hajikolaei MR, Ghorbanpour M, Gharibi D, Abdollahpour G. Serologic study on leptospiral infection in sheep in Ahvaz, southwestern Iran. Iran J Vet Res 2007; 8: 333-336.
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| Issue | Vol 3 No 3 (2011) | |
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| Leptospira Shahrekord Cattle PCR | ||
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