Prevalencia de Doenças Infecciosas em Cães Resgatados de Rinha Artigo Científico

Infectious diseases in dogs rescued during dogfighting investigations SH Cannona JK Levya SK Kirkb1 PC Crawforda CM Leuteneggerc JJ Shusterd J Liuc and R Chandrashekarc aMaddies Shelter Medicine Program Department of Small Animal Clinical Sciences College of Veterinary Medicine University of Florida Gainesville FL 32610 USA bThe American Society for the Prevention of Cruelty to Animals New York NY 10128 USA cIDEXX Laboratories Inc West Sacramento CA 95605 USA dDepartment of Health Outcomes and Policy College of Medicine University of Florida Gainesville FL 32610 USA Abstract Dogs used for dogfighting often receive minimal preventive health care and the potential for spread of infectious diseases is high The purpose of this study was to describe the prevalence of infectious diseases in dogs rescued from fighting operations to guide medical protocols for their immediate and longterm care A total of 269 pit bulltype dogs were seized in a multistate investigation Fleas were present on most dogs but few ticks were observed Testing performed at intake included packed cell volume PCV serology and PCR for vectorborne pathogens and fecal analysis The most common infections were Babesia gibsoni 39 Candidatus Mycoplasma haematoparvum 32 Mycoplasma haemocanis 30 Dirofilaria immitis 12 and Ancylostoma 23 Anemia was associated with B gibsoni infection 63 of infected dogs Odds ratio25 P0001 but not with hemotropic mycoplasmas or Ancylostoma Pit bull heritage and dogfighting are known risk factors for B gibsoni infection possibly via blood transmission from bites and vertical transmission Hemotropic mycoplasmas have a similar risk pattern Empirical care for dogs from dogfighting cases should include broadspectrum internal and external parasiticides and monitoring for anemia Dogfighting case responders should be prepared for mass screening and treatment of B gibsoni and heartworm infections and should implement Corresponding author Tel 1 352 273 8722 levyjkufledu JK Levy 1Dr Kirks current address is Cat Depot Sarasota FL 34234 USA Publishers Disclaimer This is a PDF file of an unedited manuscript that has been accepted for publication As a service to our customers we are providing this early version of the manuscript The manuscript will undergo copyediting typesetting and review of the resulting proof before it is published in its final citable form Please note that during the production process errors may be discovered which could affect the content and all legal disclaimers that apply to the journal pertain Conflict of interest statement Christian Leutenegger Jiayou Liu and Ramaswamy Chandrashekar are employed by IDEXX Laboratories None of the authors has any other financial or personal relationships that could inappropriately influence or bias the content of the paper This material was presented in part and published as an abstract at the 2014 American College of Veterinary Internal Medicine ACVIM Forum Nashville Tennessee USA 47 June 2014 HHS Public Access Author manuscript Vet J Author manuscript available in PMC 2017 October 18 Published in final edited form as Vet J 2016 May 211 6469 doi101016jtvjl201602012 Author Manuscript Author Manuscript Author Manuscript Author Manuscript protocols to prevent transmission of infectious and zoonotic diseases in the shelter and following adoption Former fighting dogs and dogs with possible dog bite scars should not be used as blood donors due to the risk of vectorborne pathogens that can escape detection and for which curative treatment is difficult to document Keywords Anemia Babesia gibsoni Canine Dogfighting Hemotropic mycoplasma Pit bull Introduction Although dogfighting is a felony offense in all 50 states and under federal law this cruel bloodsport is still entrenched in communities across the United States Growing public awareness of these crimes has led to more frequent seizures of dogs by law enforcement agencies Lockwood 2011 2013 These agencies often partner with humane organizations to provide care for seized dogs which are considered legal evidence Dogs may be housed for months in traditional or temporary animal shelters while the legal cases proceed Lockwood 2013 Fighting dogs typically receive minimal preventive care and are kept chained outside in poor conditions Multiple dogs are housed in each dog yard and are exposed to other fighting dogs during breeding training and fights The potential for spread of infectious diseases is high Since dogfighting is an underground illegal activity little is known about the infectious diseases carried by dogs from organized fighting rings Historically dogs seized in animal fighting investigations were routinely euthanized due to the belief that their heritage and training made them unsafe Recently however rescue organizations have begun assessing their health behavior and suitability for adoption in response to widespread rehoming interest Rescue operations often transport dogs around the country for adoption While this lifesaving trend is laudable there is potential risk for sending dogs harboring infectious diseases to unsuspecting owners or to regions where the infections are not currently endemic The purpose of the study reported here was to describe the prevalence of infectious diseases in dogs rescued from fighting operations This information will support an evidencebased foundation of medical protocols for biosecurity disease screening treatment and longterm followup care for this unique population Materials and methods Animals The study included 269 dogs seized from eight scenes in four states Alabama Georgia Mississippi and Texas during a federal dogfighting investigation in August and September 2013 The dogs were pit bulltype phenotypes only one of which had cropped ears Ages estimated based on dentition included neonates eight dogs 6 weeks juveniles 65 dogs 6 weeks to 59 months and adults 196 dogs 6 months A total of 52 of the dogs were female and 48 were male The dogs were triaged at the scenes vaccinated with a SC commercial rabies vaccine and transported to Florida They were housed in a climate Cannon et al Page 2 Vet J Author manuscript available in PMC 2017 October 18 Author Manuscript Author Manuscript Author Manuscript Author Manuscript controlled warehouse that served as a temporary animal shelter until the legal cases were completed Fleas were present on most dogs but few ticks were observed Ticks were not collected for species identification Upon intake to the temporary shelter dogs received intranasal vaccines containing modifiedlive Bordetella bronchiseptica parainfluenza and adenovirus2 subcutaneous vaccines containing modifiedlive distemper virus adenovirus2 parainfluenza and parvovirus and topical moxidectinimidacloprid for internal and external parasitism Adult dogs were housed individually in chainlink portable kennels Puppies were housed with littermates and nursing dams were housed with litters Adoptable dogs diagnosed with B gibsoni were treated with atovaquone 134 mgkg orally q 8 h with a fatty meal compounded into capsules Wedgewood Pharmacy and azithromycin 10 mgkg orally q 24 h for 10 days as previously described Kirk 2014 Dogs diagnosed with dirofilariasis were treated with a macrocyclic lactone monthly an oral doxycycline regimen consisting of 1 month on and 2 months off throughout their custody and with melarsomine following release from legal custody Kirk 2014 Sample collection Blood for routine health screening was collected by jugular or cephalic venipuncture into two EDTA tubes one serum separator tube and two heparinized microhematocrit tubes during examination the first week in custody Serum was harvested by centrifugation Fecal samples were collected after defecation within 4 days of intake Use of surplus blood and feces following routine health screening was approved by the University of Florida Institutional Animal Care and Use Committee Protocol 201308177 November 11 2013 Sample analysis One set of EDTA blood samples was tested onsite for Dirofilaria immitis heartworm antigen and for antibodies against Anaplasma phagocytophilum Anaplasma platys Borrelia burgdorferi Ehrlichia canis and Ehrlichia ewingii SNAP 4Dx Plus Test IDEXX Laboratories Serum was tested for B gibsoni antibodies by ELISA as described Goo et al 2008 with slight modifications In brief 96well plates were coated with fulllength rBgTRAP with 6Histag The plates were incubated with serum samples diluted to 1200 followed by color development with horseradish peroxidaseconjugated antidog IgG and optical density measured at 650 nm Samples were considered positive if the optical density was greater than the mean plus 3 standard deviations of the values from control samples collected from an Alaskan population of dogs determined to be free of B gibsoni by PCR The packed cell volume PCV was determined by centrifugation of the microhematocrit tubes For the purposes of this study anemia was defined as PCV 26 in neonates PCV 31 in juveniles and PCV 37 in adults Hoskins 2001 The second set of EDTA blood samples was tested at a commercial reference laboratory for vectorborne pathogens by realtime PCR TickVector Comprehensive RealPCR Panel Canine IDEXX Laboratories including Anaplasma phagocytophilum msp2 p44 DQ519570 A platys groEL heat shock protein AY848753 Babesia spp ssrRNA AF271082 Bartonella spp citrate synthase gene AJ439406 Mycoplasma haemocanis Cannon et al Page 3 Vet J Author manuscript available in PMC 2017 October 18 Author Manuscript Author Manuscript Author Manuscript Author Manuscript ribosomal RNA ssrRNA AF197337 Candidatus Mycoplasma haematoparvum ribosomal RNA ssrRNA AY383241 Ehrlichia canis disulfide oxidoreductase dsb gene AF403710 E ewingii disulfide oxidoreductase dsb gene AY428950 E chaffeensis disulfide oxidoreductase dsb gene AF403711 Hepatozoon canis ssrRNA AF176835 H americanum ssrRNA AF176836 Leishmania spp glycoprotein gp63 YO8156 Neorickettsia risticii ribosomal RNA 16S RNA AF184082 and Rickettsia rickettsia GroEL heat shock protein AJ293326 Realtime PCR was performed with six quality controls including quantitative PCRpositive controls PCRnegative controls negative extraction controls quantitative DNA internal sample quality control targeting the host 18S rRNA gene complex an internal positive control spiked into the lysis solution and an environmental contamination monitoring control Samples positive by PCR for Babesia spp were submitted for speciesspecific realtime PCR testing including B canis heat shock protein 70 AB248735 B canis vogeli heat shock protein 70 EF527401 B canis rossi heat shock protein 70 AB248738 B felis ITS2 AY965742 B gibsoni heat shock protein 70 AB248731 and B conradae ITS2 AY965742 All assays were designed and validated according to industry standards1 Fecal samples were processed by zinc sulfate centrifugation to screen for ova and parasites at a commercial laboratory IDEXX Laboratories and tested for Giardia spp antigen by ELISA SNAP Giardia Test IDEXX Laboratories Statistical analysis Statistical analysis was conducted in two phases In phase 1 two authors SC JL calculated the prevalence of vectorborne and intestinal pathogens For the purposes of statistical analysis dogs positive for Babesia gibsoni by either PCR or serology were considered infected Descriptive statistics for PCV and anemia were calculated and asymptotic Χ2 tests were used to test for unadjusted bivariate associations between the presence of anemia and positive results for Babesia gibsoni hemotropic mycoplasmas or Ancylostoma A value of P 005 was considered significant All calculations were made with statistical software SigmaStat for Windows 35 Systat Software Because the prevalence of vectorborne pathogens and intestinal parasitism varied among the eight investigation scenes further analysis of PCV was conducted to control for the cluster effect of scene by a third author JS The MantelHaenszel method Mantel and Haenszel 1959 was used to calculate the odds ratio OR and 95 confidence interval 95 CI for anemia defined as the ratio of the odds of anemia when the vectorborne pathogen was present to that if it was absent This also provided a Pvalue adjusted for clusters Multiple linear regression was used to compare PCV results by calculating the average difference PCV when pathogen was present minus PCV when pathogen was absent adjusted for scene and for coinfections with any of three other major pathogens A value of P 005 was considered significant All calculations in this phase were conducted with statistical software Statistical Analysis System SAS software version 93 SAS Institute 1See Applied Biosystems User Bulletin 3 httptoolsthermofishercomcontentsfsmanualscms041001pdf Accessed 26 February 2016 Cannon et al Page 4 Vet J Author manuscript available in PMC 2017 October 18 Author Manuscript Author Manuscript Author Manuscript Author Manuscript Results Pathogens Vectorborne pathogens were present in most dogs 164269 61 Table 1 The most common vectorborne pathogens were B gibsoni and hemotropic mycoplasmas the prevalence of which varied widely across the eight investigation scenes Fig 1 No dogs had evidence of infection with Anaplasma spp Bartonella spp Ehrlichia spp Hepatozoon spp Leishmania spp Neorickettsia risticii Rickettsia rickettsii or Babesia spp other than B gibsoni Results of PCR and serologic testing for B gibsoni were in agreement for 93 of dogs including 84 that were positive by both PCR and serology and 165 that were negative by both PCR and serology Two dogs were PCRpositive and serologynegative and 18 were PCRnegative and serologypositive For the purposes of statistical analysis dogs that were positive by either test 104 dogs 39 were considered infected One dog that was PCR negative and serologypositive later gave birth to B gibsoniinfected puppies This dam was retested via PCR after diagnosis in her puppies and she tested positive for B gibsoni Coinfections were more common than isolated infections A total of 76269 28 dogs were coinfected with both B gibsoni and one or both canine hemotropic mycoplasmas Table 2 There was a strong association between infection with B gibsoni and infection with Candidatus Mycoplasma haematoparvum P 00001 OR73 95 CI39136 Enteric pathogens were also commonly identified in dogs from which feces were obtained 71189 37 of dogs harbored at least one parasite Table 3 Anemia The PCV at intake ranged from 21 to 28 in neonates median 26 from 14 to 45 in juveniles median 29 and from 10 to 52 in adults median 38 In this population 28 25 neonates 4865 74 juveniles and 82196 42 adults were anemic One emaciated young adult dog had a PCV of 10 at intake The dog which had Ancylostoma on fecal analysis but was negative for vectorborne pathogens was clinically stable One week after intake and treatment with moxidectinimidacloprid the dogs PCV increased to 15 Another dog infected with B gibsoni developed a hemolytic crisis and clinical deterioration requiring a blood transfusion when PCV fell from 29 on intake to 13 3 weeks later The presence of anemia was significantly associated with B gibsoni infection in the unadjusted analysis P 0007 Table 4 and remained significant P 0001 when adjusted for the cluster effect of investigation scene Table 5 Anemia was not significantly associated with hemotropic mycoplasmas P 03 or Ancylostoma spp P 06 Median PCV was significantly lower in dogs infected solely with B gibsoni PCV 28 range 16 43 than in dogs free of vectorborne pathogens PCV 34 range 1050 P 0007 Fig 2 Median PCVs in dogs infected with hemotropic mycoplasmas or with coinfections of B gibsoni and hemotropic mycoplasmas were not significantly lower than those in dogs free of these infections P 005 Fig 2 When adjusted for investigation scene clusters and Cannon et al Page 5 Vet J Author manuscript available in PMC 2017 October 18 Author Manuscript Author Manuscript Author Manuscript Author Manuscript the presence of other vectorborne pathogens PCV was significantly decreased by a mean of 34 standard error 14 P 001 in dogs infected with B gibsoni and PCV was significantly increased by 25 standard error 12 P 003 in dogs infected with Mycoplasma haemocanis Discussion Dogs seized simultaneously from eight different scene investigations in four states during a federal investigation of organized dogfighting commonly had infectious diseases particularly B gibsoni 39 hemotropic mycoplasmas 51 Ancylostoma spp 23 Giardia spp 14 and Dirofilaria immitis 12 Most dogs kept in fighting operations in the USA are pit bulltype dogs Lockwood 2013 Although there is no universally accepted definition of a pit bull many are derived from the American Pit Bull Terrier American Staffordshire terrier Staffordshire bull terrier American bulldog and other guarding breeds Most dogs reported to be infected with B gibsoni in the USA are pit bulltype dogs or have a history of fighting with a pit bull Of 131 B gibsoniinfected dogs across the USA 122 93 were reported to be American Pit bull terriers Birkenheuer et al 2005 Of 15 B gibsoniinfected dogs of other breeds 912 had a history of having been recently bitten by another dog all of which were described as American Pit bull terriers In a study of B gibsoni infection in kennels 1429 48 American Pit bull terriers or American Staffordshire terriers from breeding operations were infected compared to only 328 11 shelter dogs of other breeds when tested by PCR Birkenheuer et al 2003 Similarly 1833 55 American Pit bull terriers from two kennels in the Southeastern USA were positive for B gibsoni by PCR whereas none of the 87 dogs of other breeds from a veterinary hospital a shelter or a Foxhound breeding kennel in the same region were infected Macintire et al 2002 In another study the prevalence of B gibsoni infection in pit bulltype dogs confiscated from dogfighting operations and housed in shelters was 34 Yeagley et al 2009 similar to the 39 prevalence in the current study The dogs with scars on the head and front limbs which are common in dogs used for organized dogfighting were 55 times more likely to be infected with B gibsoni than were dogs without scars Internationally B gibsoni is also found almost exclusively in breeds used for fighting In Romania a total of 1314 93 infected dogs were American Pit bull terriers or American Staffordshire terriers Imre et al 2013 and in Japan 3235 91 infected dogs were Tosa dogs or American Pit bull terriers Miyama et al 2005 The known competent tick vectors Haemaphysalis longicornis and Haemaphysalis bispinosa for B gibsoni in those countries are not endemic in the USA thus proposed mechanisms of transmission include other undefined biological vectors horizontal transmission through blood and saliva exchange during fighting Ayoob et al 2010 Birkenheuer et al 2005 Jefferies et al 2007 and vertical transmission from infected dams to puppies Abu et al 1973 Fukomoto et al 2005 Itoh and Itoh 1990 Approximately onethird of the dogs in this study were positive for canine hemotropic mycoplasmas by PCR The prevalence of hemotropic mycoplasmas varies widely in different regions of the world Prevalence has been reported at 06 for Mycoplasma haemocanis and 08 for Candidatus Mycoplasma haematoparvum in the USA Compton Cannon et al Page 6 Vet J Author manuscript available in PMC 2017 October 18 Author Manuscript Author Manuscript Author Manuscript Author Manuscript et al 2012 154 for hemoplasma in France Wengi et al 2008 12 for hemoplasma in Switzerland Wengi et al 2008 and 51 for Mycoplasma haemocanis and 18 for a Candidatus Mycoplasma haemominutumlike organism in Brazil de Faria Valle et al 2014 Hemotropic mycoplasma infection was not associated with anemia age or gender in one study Wengi et al 2008 whereas the presence of biological vectors old age dog bite wounds and neoplastic diseases were identified as risk factors in another study de Faria Valle et al 2014 Coinfection of hemotropic mycoplasmas and B gibsoni was reported in pit bulltype dogs rescued following Hurricane Katrina Levy et al 2011 A study conducted in Japan revealed 41 of dogs were PCR positive for Mycoplasma haemocanis Of the 913 dogs included 73 were Tosa dogs and 25 342 of those were positive for Mycoplasma haemocanis Sasaki et al 2008 These two studies combined with the current study suggest that breeds used for fighting are at increased risk for hemotropic mycoplasma infection which may share similar modes of transmission with B gibsoni Because the incidence of clinical disease attributed to hemotropic mycoplasma infections is low in immunocompetent animals and because treatment has not been shown to eliminate infection HulmeMoir et al 2010 Sykes et al 2004 the dogs in this study population were not treated for hemotropic mycoplasma infections A total of 23 of the adult dogs 12 had heartworm infection which is consistent with reports of prevalence among stray dogs in this region Bowman et al 2009 Levy et al 2011 Tzipory et al 2010 Wang et al 2014 The presence of heartworm infection created a risk for transmission to other dogs housed in the temporary shelter or in future receiving shelters or adoptive homes so all dogs were treated monthly with chemoprophylaxis Because legal custody of the dogs was not transferred to the receiving organization until after legal proceedings were completed adulticide therapy was not administered at the time of diagnosis Instead doxycycline was administered daily for 1 month on and 2 months off throughout their custody Approximately half of the dogs in this population were anemic at intake The presence of anemia was significantly associated with B gibsoni infection but not with hemotropic mycoplasmas or Ancylostoma In addition to the pathogens evaluated PCV could be affected by other unmeasured factors such as external parasitism and nutritional deficiencies Treatment at intake with moxidectinimidacloprid for flea control may have resulted in reduced detection of parasites in fecal analyses resulting in an underestimation of the impact of intestinal parasitism on PCV Despite the presence of B gibsoni or hemotropic mycoplasmas in 61 of dogs only one dog a lactating female developed a hemolytic crisis requiring a blood transfusion This dog was emaciated on intake and was actively nursing three puppies Three weeks after admission into the temporary shelter the dog was lethargic anorexic and had pale mucous membranes A complete blood count and differential revealed regenerative anemia with PCV of 13 PCV at intake was 29 no blood parasites were observed microscopically although this dog was positive for B gibsoni by both PCR and serology at intake Results of infectious disease screening of this population indicate that dogs seized in dogfighting investigations are at increased risk of infection with Babesia gibsoni and hemotropic mycoplasmas In addition fleas dirofilariasis Giardia and Ancylostoma were Cannon et al Page 7 Vet J Author manuscript available in PMC 2017 October 18 Author Manuscript Author Manuscript Author Manuscript Author Manuscript also common Thus caregivers in dogfighting cases should be alerted to risk of infections observe dogs for clinical signs and consider mass screening for vectorborne pathogens Pregnant dogs lactating dogs and puppies are often seized during dogfighting investigations these populations may be at greater risk of exhibiting clinical signs from infections and thus warrant closer monitoring for anemia Dogs seized from dogfighting investigations should not be used as blood donors due to high risk of subclinical infections with vectorborne pathogens Blood donor candidates that have suspected fighting breed heritage or scars from dog bites should receive repeated screening by both PCR and serology to rule out B gibsoni in addition to routine disease screening panels Wardrop et al 2005 prior to donating blood Conclusions Pit bull heritage and dogfighting are known risk factors for B gibsoni infection possibly via blood or saliva transmission from bites and vertical transmission Hemotropic mycoplasmas appear to follow a similar risk pattern suggesting a common mode of transmission Mild to moderate anemia was common in seized dogs but many anemia cases were not associated with documented infections or parasitism other than fleas Empirical care for all dogs seized in dogfighting investigations should include broadspectrum internal and external parasiticides routine vaccinations and monitoring for anemia Dogfighting case responders should be prepared for mass screening and treatment of B gibsoni and heartworm infections and should implement protocols to prevent transmission of infectious and zoonotic diseases in the shelter and following adoption Former fighting dogs and dogs with possible dog bite scars should not be used as blood donors due to the risk of vectorborne pathogens that can escape detection and for which curative treatment is difficult to document Acknowledgments This study was supported by grants from Maddies Fund and by the Lois Kugler Small Animal Research Trust PCR testing B gibsoni serology and SNAP 4DxPlus test kits were contributed by IDEXX Laboratories This work was supported in part by the NIHNCATS Clinical and Translational Science Award to the University of Florida UL1 TR000064 References Abu M Hara I Naito I Shibauchi O Babesia infections in puppies probably due to transplacental transmission Journal of Veterinary Medicine 1973 609203206 Ayoob AL Hackner SG Prittie J Clinical management of canine babesiosis Journal of Veterinary Emergency and Critical Care 2010 207789 PubMed 20230437 Birkenheuer AJ Correa MT Levy MG Breitschwerdt EB Geographic distribution of babesiosis among dogs in the United States and association with dog bites 150 cases 20002003 Journal of the American Veterinary Medical Association 2005 227942947 PubMed 16190594 Birkenheuer AJ Levy MG Stebbins M Poore M Breitschwerdt E Serosurvey of antiBabesia antibodies in stray dogs and American pit bull terriers and American staffordshire terriers from North Carolina Journal of the American Animal Hospital Association 2003 39551557 PubMed 14736721 Bowman D Little SE Lorentzen L Shields J Sullivan MP Carlin EP Prevalence and geographic distribution of Dirofilaria immitis Borrelia burgdorferi Ehrlichia canis and Anaplasma phagocytophilum in dogs in the United States Results of a national clinicbased serologic survey Veterinary Parasitology 2009 160138148 PubMed 19150176 Cannon et al Page 8 Vet J Author manuscript available in PMC 2017 October 18 Author Manuscript Author Manuscript Author Manuscript Author Manuscript Compton SM Maggi RG Breitschwerdt EB Candidatus Mycoplasma Haematoparvum and Mycoplasma haemocanis infections in dogs from the United States Comparative Immunology Microbiology and Infectious Diseases 2012 35557562 de Faria Valle S Messick JB Pires dos Santos A Kreutz LC Blatt Duda NC Machado G Corbellini LG Biondo AW Diaz Gonzalez FH Identification occurrence and clinical findings of canine hemoplasmas in southern Brazil Comparative Immunology Microbiology and Infectious Diseases 2014 37259265 Fukumoto S Suzuki H Igarashi I Xuan X Fatal experimental transplacental Babesia gibsoni infections in dogs International Journal for Parasitology 2005 3510311035 PubMed 15979628 Goo YK1 Jia H Aboge GO Terkawi MA Kuriki K Nakamura C Kumagai A Zhou J Lee EG Nishikawa Y Igarashi I Fujisaki K Xuan X Babesia gibsoni Serodiagnosis of infection in dogs by an enzymelinked immunosorbent assay with recombinant BgTRAP Experimental Parasitology 2008 118555560 PubMed 18155197 Hoskins JD Veterinary Pediatrics Dogs and Cats from Birth to Six Months 3 Saunders Philadelphia PA 2001 p 594 HulmeMoir KL Barker EN Stonelake A Helps CR Tasker S Use of realtime quantitative polymerase chain reaction to monitor antibiotic therapy in a dog with naturally acquired Mycoplasma haemocanis infection Journal of Veterinary Diagnostic Investigation 2010 22582 587 PubMed 20622229 Imre M Farkas R Ilie MS Imre K Darabus G Survey of babesiosis in symptomatic dogs from Romania occurrence of Babesia gibsoni associated with breed Ticks and Tickborne Diseases 2013 4500502 PubMed 23994336 Itoh N Itoh S A case of canine babesiosis possibly developed by transplacental infection Journal of the Japanese Veterinary Medical Association 1990 43275276 Jefferies R Ryan UM Jardine J Broughton DK Robertson ID Irwin PJ Blood Bull Terriers and Babesiosis further evidence for direct transmission of Babesia gibsoni in dogs Australian Veterinary Journal 2007 85459463 PubMed 17970851 Kirk S Efficacy of azithromycin and compounded atovaquone for treatment of Babesia gibsoni in a largescale dogfighting case Proceedings of the American College of Veterinary Internal Medicine ACVIM Forum Nashville Tennessee USA 47 June 2014 2014 Levy JK Lappin MR Glaser AL Birkenheuer AJ Anderson TC Edinboro CH Prevalence of infectious diseases in cats and dogs rescued following Hurricane Katrina Journal of the American Veterinary Medical Association 2011 238311317 PubMed 21281213 Lockwood R Dogfighting Toolkit for Law Enforcement Addressing Dogfighting in Your Community US Department of Justices Office of Community Policing Services and American Society for Prevention of Cruelty to Animals ASPCA 2011 Lockwood R Animal Fighting In Miller L Zawistowski S editors Shelter Medicine for Veterinarians and Staff 2 John Wiley Sons Inc Ames Iowa USA 2013 p 441452 Macintire DK Boudreaux MK West GD Bourne C Wright JC Conrad PA Babesia gibsoni infection among dogs in the southeastern United States Journal of the American Veterinary Medical Association 2002 220325329 PubMed 11829262 Mantel N Haenszel W Statistical aspects of the analysis of data from retrospective studies of disease Journal of the National Cancer Institute 1959 22719748 PubMed 13655060 Miyama T Sakata Y Shimada Y Ogino S Watanabe M Itamoto K Okuda M Verdida RA Zuan Z Nagasawa H et al Epidemiological Survey of Babesia gibsoni Infection in Dogs in Eastern Japan The Journal of Veterinary Medical Science 2005 67467471 PubMed 15942130 Sasaki M Ohta K Matsuu A Hirata H Ikadai H Oyamada T A molecular survey of Mycoplasma haemocanis in dogs and foxes in Aomori Prefecture Japan The Journal of Protozoology Research 2008 185760 Sykes JE Bailiff NL Ball LM Foreman O George JW Fry MM Identification of a novel hemotropic mycoplasma in a splenectomized dog with hemic neoplasia Journal of the American Veterinary Medical Association 2004 22419461951 PubMed 15230449 Cannon et al Page 9 Vet J Author manuscript available in PMC 2017 October 18 Author Manuscript Author Manuscript Author Manuscript Author Manuscript Tzipory N Crawford PC Levy JK Prevalence of Dirofilaria immitis Ehrlichia canis and Borrelia burgdorferi in pet dogs racing greyhounds and shelter dogs in Florida Veterinary Parasitology 2010 171136139 PubMed 20399018 Wang D Bowman DD Brown HE Harrington LC Kaufman PE McKay T Nelson CT Sharp JL Lund R Factors influencing US canine heartworm Dirofilaria immitis prevalence Parasites Vectors 2014 7118 PubMed 24411014 Wardrop KJ Reine N Birkenheuer A Hale A Hohenhaus A Crawford C Lappin MR Canine and feline blood donor screening for infectious disease Journal of Veterinary Internal Medicine 2005 19135142 PubMed 15715062 Wengi N Willi B Boretti FS Cattori V Riond B Meli ML Reusch CE Lutz H HofmannLehmann R Realtime PCRbased prevalence study infection follow up and molecular characterization of canine hemotropic mycoplasmas Veterinary Microbiology 2008 126132141 PubMed 17656047 Yeagley TJ Reichard MV Hempstead JE Allen KE Parsons LM White MA Little SE Meinkoth JH Detection of Babesia gibsoni and the canine small Babesia Spanish isolate in blood samples obtained from dogs confiscated from dogfighting operations Journal of the American Veterinary Medical Association 2009 235535539 PubMed 19719443 Cannon et al Page 10 Vet J Author manuscript available in PMC 2017 October 18 Author Manuscript Author Manuscript Author Manuscript Author Manuscript Highlights Babesia gibsoni and hemotropic mycoplasmas are common in dogs in fighting operations Anemia is common and is most strongly associated with B gibsoni infection Fighting dogs should not be blood donors due to the risk of transmitting pathogens Cannon et al Page 11 Vet J Author manuscript available in PMC 2017 October 18 Author Manuscript Author Manuscript Author Manuscript Author Manuscript Fig 1 Prevalence of Babesia gibsoni A Candidatus Mycoplasma haematoparvum B and Mycoplasma haemocanis C infection in dogs from each of the eight scenes in a federal dogfighting investigation and prevalence from all scenes combined n number of dogs seized from each scene Cannon et al Page 12 Vet J Author manuscript available in PMC 2017 October 18 Author Manuscript Author Manuscript Author Manuscript Author Manuscript Fig 2 Packed cell volume PCV in dogs with and without vectorborne pathogen infections Boxes represent the 25th and 75th percentiles lines indicate the medians whiskers represent the 10th and 90th percentiles and circles indicate outliers The number of dogs in each column corresponds to the data presented in Table 2 Median PCV in dogs solely infected with B gibsoni Bg was significantly lower than in dogs free of infections Neg Neg negative for B gibsoni and hemotropic mycoplasmas Bg Babesia gibsoni MHc Mycoplasma haemocanis CMhp Candidatus Mycoplasma haematoparvum P0007 Cannon et al Page 13 Vet J Author manuscript available in PMC 2017 October 18 Author Manuscript Author Manuscript Author Manuscript Author Manuscript Author Manuscript Author Manuscript Author Manuscript Author Manuscript Cannon et al Page 14 Table 1 Prevalence of vectorborne pathogens in 269 dogs seized from eight scenes in a federal dogfighting investigation Test method Dogs tested n Dogs positive n Percent positive Babesia gibsoni Serology 261 102 38 Babesia gibsoni PCR 269 86 32 Candidatus Mycoplasma haematoparvum PCR 269 86 32 Mycoplasma haemocanis PCR 269 80 30 Dirofilaria immitis1 Serology 196 23 12 Anaplasma spp PCR 269 0 0 Anaplasma spp Serology 269 0 0 Bartonella spp PCR 269 0 0 Ehrlichia spp PCR 269 0 0 Ehrlichia spp Serology 269 0 0 Hepatozoon spp PCR 269 0 0 Leishmania spp PCR 269 0 0 Neorickettsia risticii PCR 269 0 0 Rickettsia spp PCR 269 0 0 Borrelia burgdorferi Serology 269 0 0 1Testing for D immitis was performed only on adult dogs 6 months and older Vet J Author manuscript available in PMC 2017 October 18 Author Manuscript Author Manuscript Author Manuscript Author Manuscript Cannon et al Page 15 Table 2 Babesia gibsoni and hemotropic mycoplasma coinfections in 269 dogs seized from eight scenes in a federal dogfighting investigation Babesia gibsoni infection includes dogs that had positive PCR andor serology test results n104 Dogs positive n Percent positive MHc only 37 14 Bg only 28 10 CMhp only 15 6 Bg CMhp 41 15 Bg MHc CMhp 22 8 Bg MHc 13 5 MHc CMhp 8 3 MHc Mycoplasma haemocanis Bg Babesia gibsoni CMhp Candidatus Mycoplasma haematoparvum Vet J Author manuscript available in PMC 2017 October 18 Author Manuscript Author Manuscript Author Manuscript Author Manuscript Cannon et al Page 16 Table 3 Prevalence of intestinal parasites in 189 dogs seized from eight scenes in a federal dogfighting investigation Test method Dogs tested n Dogs positive n Percent positive Giardia spp ELISA 162 22 14 Ancylostoma spp Zinc sulfate centrifugation 189 44 23 Isospora spp Zinc sulfate centrifugation 189 9 5 Trichuris vulpis Zinc sulfate centrifugation 189 6 3 Toxocara spp Zinc sulfate centrifugation 189 3 2 Vet J Author manuscript available in PMC 2017 October 18 Author Manuscript Author Manuscript Author Manuscript Author Manuscript Cannon et al Page 17 Table 4 Anemia in dogs with Babesia gibsoni Ancylostoma or hemotropic mycoplasma For the purposes of this study anemia was defined as packed cell volume PCV 26 in neonates PCV 31 in juveniles and PCV 37 in adults Pathogen Infection status n Anemia n Median PCV PCV range P Babesia gibsoni Uninfected 165 61 67 41 37 1050 Infected 104 39 65 63 34 1652 00007 Ancylostoma spp Uninfected 145 77 62 43 37 1452 Infected 44 23 23 52 35 1047 03014 Candidatus Mycoplasma haematoparvum Uninfected 183 68 90 49 34 1050 Infected 86 32 42 49 37 2052 1000 Mycoplasma haemocanis Uninfected 189 70 98 52 34 1052 01829 Infected 80 30 34 43 38 1949 Vet J Author manuscript available in PMC 2017 October 18 Author Manuscript Author Manuscript Author Manuscript Author Manuscript Cannon et al Page 18 Table 5 Risk of anemia in dogs with Babesia gibsoni Ancylostoma spp or hemotropic mycoplasmas adjusted for investigation scene Estimated OR 95 CI P twosided Babesia gibsoni 25 1543 0001 Ancylostoma spp 15 0831 023 Candidatus Mycoplasma haematoparvum 09 0515 063 Mycoplasma haemocanis 07 0413 026 OR odds ratio for anemia presentabsent CI confidence Interval Vet J Author manuscript available in PMC 2017 October 18