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Bacterial superinfection with venereal pathogens subsequent to topical therapy for Taylorella equigenitalis in stallions
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9th ICEID Abstracts / Journal of Equine Veterinary Science 32 (2012) S3-S95
virus detection by cell culture or PCR. While most of the recent studies were described in Argentina, the need to improve the presence of such virus in other countries still exists. In France, between March 2002 and March 2012, thirteen swabs were collected from the genital area of 4 stallions and 9 mares for EHV-3 investigation after an ECE suspicion. DNA was extracted using the QIAamp DNA mini kit. We used consensual herpes PCR method associated with sequencing from 2002 to 2005 and had developed a qPCR method (using SybrGreen) for samples tested from 2006 to 2012. The real-time PCR was tested against EHV-1, EHV-4, EHV-2, EHV-5 and AHV-5 and a range of pathogens often isolated from the equine reproductive tract. These included Equine viral arteritis virus, Taylorella equigenitalis, Klebsiella pneumoniae and Streptococcus zooepidemicus. The analytical sensitivity of the real-time PCR was compared to TCID50. The qPCR was validated for specificity and presented a sensitivity of 3.2 TCID50/50ml. Ten cases were confirmed by PCR analysis between March 2002 and March 2012 in France out of thirteen examinations. Six mares and four stallions were declared positive after confirmation of the presence of EHV-3 DNA by classical PCR (3 cases) and real-time PCR (7 cases). Different horse breeds were concerned by the disease (Comtois, Welsh Pony, Shetland pony, French Saddle horse, Thoroughbred) and were all between 3 and 27 years old. All cases are from sporadic infections that were observed in different French departments (35, 71, 72, 69, and 54) except that occurred in May 2005 which involved seven horses; even if only one stallion and one mare were confirmed positive by PCR. During this episode there is evidence of contamination of mare by a stallion without noticeable lesions. Our study presents a new efficient real-time PCR test which could be used for confirmation of ECE diagnosis and demonstrate that the virus circulate in France.
Bacterial superinfection with venereal pathogens subsequent to topical therapy for Taylorella equigenitalis in stallions M.L. Schulman, B. Keys, C.E. May, and A.J. Guthrie Faculty of Veterinary Science, University of Pretoria, Onderstepoort, South Africa
Venereal pathogens including Klebsiella pneumoniae and Pseudomonas aeruginosa may establish asymptomatic carrier status and associated subfertility after colonisation of the external genitalia of stallions due to a disturbance of normal commensal microflora subsequent to antimicrobial and disinfectant therapy [1]. An outbreak of Taylorella equigenitalis among the resident stallions at the Lipizzaner Centre in South Africa was managed by a currently-accepted treatment protocol of topical antibacterials and disinfectants [2, 3]. A predisposition to infection has been reported following similar therapy for T. equigenitalis [4]. A controlled study aimed to observe the effects of treatment on the bacterial microflora of the external genitalia of the T. equigenitalis-infected stallions. The protocol consisted of topical treatment of the penis
and prepuce of positive stallions with surfactant (5% docusate sodium solution), disinfectant (4% chlorhexidine gluconate solution) and either one of two topical antimicrobials (0.2% nitrofurazone or 1% silver sulphadiazine). Twenty one positive stallions randomly allocated to either of two treatment groups, Group 1 (nitrofurazone, n¼ 11) or Group 2 (silver sulphadiazine, n¼ 10), were treated daily for > 9d until swabbing negative for T. equigenitalis on daily qPCR, confirmed by bacteriology after 3 weeks. The mean interval (and range) for successful treatment overall was 15.1 d (9-28 d). Nine negative stallions quarantined without treatment served as controls. Four weeks after the last treatment, three sites (distal urethra, urethral fossa including sinus and lamina interna) were swabbed in all 30 stallions for bacteriology. The effects of treatment and antibacterial choice on bacterial cultures and predilection sites for bacterial pathogens were compared using standard statistical methods. Statistical significance in all cases was set at P < 0.05. There was a significant effect on the colonisation by bacterial pathogens following either treatment with a 54.5% and 80.0% occurrence in the stallions of Group 1 and 2 respectively compared with 11.1% in the Controls. The culture sites showed a similar distribution between the two organisms and with an occurrence of 36.0% (urethral fossa and sinus), 22.0% (lamina) and 11.1% (urethra). Current treatment protocols for T. equigenitalis significantly increase the probability for subsequent colonisation of the external genitalia by potential bacterial pathogens.
References [1] Lu K, Morressey PR. Infectious Diseases in Breeding Stallions. Clin Tech Equine Pract 2007;6:285-90. [2] Erdman MM, Creekmore LH, Fox PE, Pelzel AM, Porter-Spalding BA, Aalsburg AM, Cox LK, Morningstar-Shaw BR, Crom RL. Diagnostic and epidemiologic analysis of the 2008-2010 investigation of a multi-year outbreak of contagious equine metritis in the United States. Prev Vet Med 2011;101:219-28. [3] Kristula MA, Smith BI. Diagnosis and treatment of four stallions, carriers of the contagious metritis organism: case report. Therio 2004;61:595-601. [4] Swerczek TW. Aggravation of strangles, equine clostridial typhlocolitis (colitis X) and bacterial venereal diseases in the horse by antibacterial drugs. Proc 25th Ann Conv AAEP; 1979:305-11.
Effect of acyclovir, ganciclovir and cidofovir on equid herpesvirus 3 in vitro A. Vissani 1, S. Tordoya 1, L. Becerra 1, E. Thiry 2, and M. Barrandeguy 1 1 Instituto de Virología, CICVyA, INTA-Castelar, CC25, 1712 Castelar, Buenos Aires, Argentina, 2 Veterinary Virology and Animal Viral Diseases, Department of Infectious and Parasitic Diseases, Faculty of Veterinary Medicine, University of Liege, B-4000 Liege, Belgium Equine Coital Exanthema (ECE), which is caused by equid herpesvirus 3 (EHV-3), is a venereal, highly contagious disease, characterized by the formation of papules, vesicles, pustules and ulcers on the external genitalia of mares and stallions. EHV-3 infection is endemic in horses
9th ICEID Abstracts / Journal of Equine Veterinary Science 32 (2012) S3-S95
virus detection by cell culture or PCR. While most of the recent studies were described in Argentina, the need to improve the presence of such virus in other countries still exists. In France, between March 2002 and March 2012, thirteen swabs were collected from the genital area of 4 stallions and 9 mares for EHV-3 investigation after an ECE suspicion. DNA was extracted using the QIAamp DNA mini kit. We used consensual herpes PCR method associated with sequencing from 2002 to 2005 and had developed a qPCR method (using SybrGreen) for samples tested from 2006 to 2012. The real-time PCR was tested against EHV-1, EHV-4, EHV-2, EHV-5 and AHV-5 and a range of pathogens often isolated from the equine reproductive tract. These included Equine viral arteritis virus, Taylorella equigenitalis, Klebsiella pneumoniae and Streptococcus zooepidemicus. The analytical sensitivity of the real-time PCR was compared to TCID50. The qPCR was validated for specificity and presented a sensitivity of 3.2 TCID50/50ml. Ten cases were confirmed by PCR analysis between March 2002 and March 2012 in France out of thirteen examinations. Six mares and four stallions were declared positive after confirmation of the presence of EHV-3 DNA by classical PCR (3 cases) and real-time PCR (7 cases). Different horse breeds were concerned by the disease (Comtois, Welsh Pony, Shetland pony, French Saddle horse, Thoroughbred) and were all between 3 and 27 years old. All cases are from sporadic infections that were observed in different French departments (35, 71, 72, 69, and 54) except that occurred in May 2005 which involved seven horses; even if only one stallion and one mare were confirmed positive by PCR. During this episode there is evidence of contamination of mare by a stallion without noticeable lesions. Our study presents a new efficient real-time PCR test which could be used for confirmation of ECE diagnosis and demonstrate that the virus circulate in France.
Bacterial superinfection with venereal pathogens subsequent to topical therapy for Taylorella equigenitalis in stallions M.L. Schulman, B. Keys, C.E. May, and A.J. Guthrie Faculty of Veterinary Science, University of Pretoria, Onderstepoort, South Africa
Venereal pathogens including Klebsiella pneumoniae and Pseudomonas aeruginosa may establish asymptomatic carrier status and associated subfertility after colonisation of the external genitalia of stallions due to a disturbance of normal commensal microflora subsequent to antimicrobial and disinfectant therapy [1]. An outbreak of Taylorella equigenitalis among the resident stallions at the Lipizzaner Centre in South Africa was managed by a currently-accepted treatment protocol of topical antibacterials and disinfectants [2, 3]. A predisposition to infection has been reported following similar therapy for T. equigenitalis [4]. A controlled study aimed to observe the effects of treatment on the bacterial microflora of the external genitalia of the T. equigenitalis-infected stallions. The protocol consisted of topical treatment of the penis
and prepuce of positive stallions with surfactant (5% docusate sodium solution), disinfectant (4% chlorhexidine gluconate solution) and either one of two topical antimicrobials (0.2% nitrofurazone or 1% silver sulphadiazine). Twenty one positive stallions randomly allocated to either of two treatment groups, Group 1 (nitrofurazone, n¼ 11) or Group 2 (silver sulphadiazine, n¼ 10), were treated daily for > 9d until swabbing negative for T. equigenitalis on daily qPCR, confirmed by bacteriology after 3 weeks. The mean interval (and range) for successful treatment overall was 15.1 d (9-28 d). Nine negative stallions quarantined without treatment served as controls. Four weeks after the last treatment, three sites (distal urethra, urethral fossa including sinus and lamina interna) were swabbed in all 30 stallions for bacteriology. The effects of treatment and antibacterial choice on bacterial cultures and predilection sites for bacterial pathogens were compared using standard statistical methods. Statistical significance in all cases was set at P < 0.05. There was a significant effect on the colonisation by bacterial pathogens following either treatment with a 54.5% and 80.0% occurrence in the stallions of Group 1 and 2 respectively compared with 11.1% in the Controls. The culture sites showed a similar distribution between the two organisms and with an occurrence of 36.0% (urethral fossa and sinus), 22.0% (lamina) and 11.1% (urethra). Current treatment protocols for T. equigenitalis significantly increase the probability for subsequent colonisation of the external genitalia by potential bacterial pathogens.
References [1] Lu K, Morressey PR. Infectious Diseases in Breeding Stallions. Clin Tech Equine Pract 2007;6:285-90. [2] Erdman MM, Creekmore LH, Fox PE, Pelzel AM, Porter-Spalding BA, Aalsburg AM, Cox LK, Morningstar-Shaw BR, Crom RL. Diagnostic and epidemiologic analysis of the 2008-2010 investigation of a multi-year outbreak of contagious equine metritis in the United States. Prev Vet Med 2011;101:219-28. [3] Kristula MA, Smith BI. Diagnosis and treatment of four stallions, carriers of the contagious metritis organism: case report. Therio 2004;61:595-601. [4] Swerczek TW. Aggravation of strangles, equine clostridial typhlocolitis (colitis X) and bacterial venereal diseases in the horse by antibacterial drugs. Proc 25th Ann Conv AAEP; 1979:305-11.
Effect of acyclovir, ganciclovir and cidofovir on equid herpesvirus 3 in vitro A. Vissani 1, S. Tordoya 1, L. Becerra 1, E. Thiry 2, and M. Barrandeguy 1 1 Instituto de Virología, CICVyA, INTA-Castelar, CC25, 1712 Castelar, Buenos Aires, Argentina, 2 Veterinary Virology and Animal Viral Diseases, Department of Infectious and Parasitic Diseases, Faculty of Veterinary Medicine, University of Liege, B-4000 Liege, Belgium Equine Coital Exanthema (ECE), which is caused by equid herpesvirus 3 (EHV-3), is a venereal, highly contagious disease, characterized by the formation of papules, vesicles, pustules and ulcers on the external genitalia of mares and stallions. EHV-3 infection is endemic in horses