P. Cynda Crawford

Influenza A Virus (H3N8) in Dogs with Respiratory Disease, Florida

Genetic and antigenic characterization established the uniqueness of this virus circulating in dogs.

Experimental Infection of Neonatal Foals with Rhodococcus equi Triggers Adult-Like Gamma Interferon Induction

ABSTRACT Rhodococcus equi is a facultative intracellular pathogen that causes pneumonia in young foals but does not induce disease in immunocompetent adult horses. Clearance of R. equi depends mainly on gamma interferon (IFN-γ) production by T lymphocytes, whereas the predominance of interleukin 4 (IL-4) is detrimental. Young foals, like neonates of many other species, are generally deficient in the ability to produce IFN-γ. The objective of this study was to compare the cytokine profiles, as well as cell-mediated and antibody responses, of young foals to those of adult horses following intrabronchial challenge with R. equi. The lymphoproliferative responses of bronchial lymph node (BLN) cells to concanavalin A were significantly higher in foals than in adult horses. In contrast, adult horses had significantly higher lymphoproliferative responses to R. equi antigens than did foals. Infected foals had significantly lower IL-4 mRNA expression but significantly higher IFN-γ expression and IFN-γ/IL-4 ratio in R. equi-stimulated BLN lymphocytes than did infected adults. Infection with R. equi in foals resulted in a significant increase in the percentage of T lymphocytes and CD4+ T lymphocytes in bronchoalveolar lavage fluid in association with a significant decrease in the percentage of these cell populations in BLNs. Infection of foals also resulted in a marked increase in serum immunoglobulin Ga (IgGa) and IgGb levels, resulting in concentrations in serum that were significantly higher than those of adult horses. This study demonstrates that the immune response to R. equi in foals is not biased toward IL-4 and is characterized by the predominant induction of IFN-γ.

Comparison of intratesticular injection of zinc gluconate versus surgical castration to sterilize male dogs

OBJECTIVE To compare castration of dogs by use of intratesticular injection of zinc gluconate with traditional surgical procedures in terms of acceptance by pet owners, ease of use, and short-term outcomes on Isabela Island of the Galápagos Islands. ANIMALS 161 privately owned male dogs admitted to a neuter program. PROCEDURES Medical records of male dogs neutered during a 4-week animal control campaign were reviewed to collect information regarding signalment, method of castration, complication rate, and treatment outcomes. RESULTS Of the 161 dogs admitted for castration, 58 were surgically castrated and 103 were treated with zinc gluconate. Dogs were returned to their owners for observation following castration. Wound dehiscence occurred in 2 skin incisions, representing 3.4% of the 58 dogs that underwent bilateral orchiectomy. Necrotizing zinc-gluconate injection-site reactions occurred in 4 dogs receiving injection volumes near the maximum label dose (0.8 to 1.0 mL), representing 3.9% of the zinc-gluconate procedures. Surgical wound complications were treated by superficial wound debridement and resuturing, in contrast to zinc-gluconate injection-site reactions, which all required orchiectomy and extensive surgical debridement, including scrotal ablation in 2 dogs. CONCLUSIONS AND CLINICAL RELEVANCE Low cost, ease of use, and cultural acceptance of a castration technique that does not require removal of the testes make zinc gluconate a valuable option for large-scale use in dogs, particularly in remote locations lacking sophisticated clinical facilities or skilled surgeons and staff. Further investigation is needed to identify risk factors in dogs for adverse reactions to zinc gluconate and to develop strategies for avoidance.

Prevalence of protective antibody titers for canine distemper virus and canine parvovirus in dogs entering a Florida animal shelter.

OBJECTIVE To determine the proportion of dogs entering an animal shelter with protective antibody titers (PATs) for canine distemper virus (CDV) and canine parvovirus (CPV) and identify factors associated with having a PAT. DESIGN Cross-sectional study. ANIMALS 431 dogs admitted to an open-admission municipal animal shelter in north central Florida with a history of infectious disease outbreaks. PROCEDURES Blood was collected from dogs on the day of admission to the shelter. Antibody titers for CDV and CPV were measured by virus neutralization and hemagglutination inhibition, respectively. Age, sex, neuter status, address of origin, source (stray or previously owned), health status (healthy or not healthy), and outcome (adoption, euthanasia, or reclaimed by owner) data were also collected. RESULTS Overall, 64.5% (278/431) of dogs had insufficient titers for antibodies against CDV, CPV, or both. A total of 153 (35.5%) dogs had PATs for both CDV and CPV, 33 (7.7%) had PATs for CDV but not CPV, 136 (31.5%) had PATs for CPV but not CDV, and 109 (25.3%) did not have PATs for either virus. Older dogs were more likely to have PATs for CDV and CPV. Neutered dogs were more likely to have PATs for CDV. Factors not associated with having a PAT included source, health status, and type of community from which the dog originated. CONCLUSIONS AND CLINICAL RELEVANCE Most dogs had insufficient antibody titers for CDV, CPV, or both at the time of admission to the animal shelter. Findings support current guidelines recommending vaccination of all dogs immediately upon admission to shelters, regardless of source or physical condition.

Immunoglobulin concentrations in feline colostrum and milk, and the requirement of colostrum for passive transfer of immunity to neonatal kittens.

The purpose of this study was to clarify whether cats have a colostral and milk phase of lactation differentiated by concentrations of immunoglobulins, and whether colostrum ingestion by newborn kittens is essential for optimal transfer of passive immunity. Milk from specific pathogen-free queens was analyzed for IgG and IgA concentrations from parturition through 6 weeks of lactation. Serum IgG and IgA concentrations from birth through 8 weeks of age were determined for colostrum-fed kittens, colostrum-deprived kittens that were fed a milk replacer, and colostrum-deprived kittens that were fostered onto queens in the milk phase of lactation. The total IgG and IgA concentrations in milk were significantly higher on the day of parturition than on day 7 of lactation, indicating cats do have a colostral phase of lactation. The predominant immunoglobulin in both colostrum and milk was IgG. The serum IgG concentrations in colostrum-deprived kittens fostered on queens in the milk phase of lactation were similar to colostrum-deprived kittens fed a milk replacer, and the concentrations were significantly lower than in colostrum-fed kittens for the first 4 weeks of life. The serum IgA concentrations in both colostrum-deprived groups were significantly lower than colostrum-fed kittens on day 2 after parturition, but were similar thereafter. Colostrum-deprived kittens fostered onto queens in the milk phase of lactation had failure of passive transfer of maternal antibodies. Protective concentrations of immunoglobulins can be restored in kittens with failure of passive transfer of immunity by parenteral administration of adult cat serum, but not by fostering on queens in mid-lactation.

Effects of two commercially available immunostimulants on leukocyte function of foals following ex vivo exposure to Rhodococcus equi

The objective of this study was to determine the effect of immunostimulants on neutrophil, macrophage, and lymphocyte function following ex vivo exposure to Rhodococcus equi. Eighteen foals were randomly assigned to one of 3 treatment groups. Treatment consisted of inactivated Propionibacterium acnes (PA), inactivated parapoxvirus ovis (PPVO), or saline (control) administered on days 0 (7 days of age), 2, and 8. Bronchoalveolar lavage (BAL) fluid and blood were collected on days 0 (baseline), 12, 24 and 36. Intracellular replication of R. equi in macrophages, cytokine induction by R. equi-infected macrophages, phagocytic and oxidative burst activity of neutrophils, lymphoproliferative responses, and cytokine induction of proliferating lymphocytes were measured. Neutrophils from foals treated with PPVO had significantly greater ability to phagocytize R. equi and undergo oxidative burst on day 12 and day 24 compared to baseline values. On day 24, foals treated with PPVO had significantly greater phagocytosis and oxidative burst than foals treated with PA. Treatment with PA resulted in significantly less intracellular proliferation of R. equi within monocyte-derived macrophages on day 12 compared to control foals. The ability of R. equi to replicate in BAL macrophages decreased significantly (P=0.005) with time with lower replication in BAL macrophages of older foals compared to younger foals, regardless of treatment. On day 12, TNF-α induction in monocyte-derived macrophages and IL-12 p40 induction in BAL macrophages infected with R. equi was significantly higher in foals treated with PPVO than in controls. Lymphoproliferative responses and IFN-γ induction were not significantly different between groups.

Prevalence of and exposure factors for seropositivity to H3N8 canine influenza virus in dogs with influenza-like illness in the United States.

OBJECTIVE To estimate the seroprevalence of antibodies against H3N8 canine influenza virus (CIV) in a population of US dogs with influenza-like illness (ILI) and to identify factors associated with seropositivity. DESIGN Cross-sectional study. ANIMALS 1,268 pet and shelter dogs with ILI in 42 states. PROCEDURES Serum samples collected from dogs from 2005 through June 2009 were tested for H3N8 CIV antibodies with a hemagglutination inhibition assay. Intrinsic factors (age, breed, and sex), extrinsic factors (dogs housed in a shelter facility, boarding kennel, or other setting), and geographic region (southwest, west, Midwest, southeast, and northeast) were compared between seropositive and seronegative dogs to identify variables associated with seropositivity. RESULTS Most (750/1,268 [59%]) dogs in the study were from Colorado, Florida, or New York. The overall seroprevalence of antibodies against H3N8 CIV was 49% (618/1,268 dogs; 95% confidence interval, 46% to 51%). The annual prevalence of H3N8 CIV seropositivity increased from 2005 (44%) to 2006 (53%) and 2007 (62%), then decreased in 2008 (38%) and 2009 (15%). The likelihood of H3N8 CIV seropositivity was associated with geographic region (southeast during 2005, west and northeast during 2006 and 2007, and northeast during 2008) and exposure setting (dogs housed in a shelter facility or boarding kennel during 2005 and 2006). CONCLUSIONS AND CLINICAL RELEVANCE Results of this study suggested there is a need for continued surveillance for H3N8 CIV infection in dogs in the United States and that personnel in communal dog-housing facilities should formulate, implement, and evaluate biosecurity protocols to reduce the risk of CIV transmission among dogs.

Prevalence of Dirofilaria immitis, Ehrlichia canis, and Borrelia burgdorferi in pet dogs, racing greyhounds, and shelter dogs in Florida

Arthropod vectors of canine infectious diseases are present throughout Florida. Since crowded housing has the potential to bring vectors and infected dogs into close proximity, it is possible that prevalence of infection is higher in intensely housed dogs. In this study, the seroprevalence of Dirofilaria immitis, Ehrlichia canis, and Borrelia burgdorferi in dogs residing in two types of intensive housing, greyhound kennels and animal shelters, was compared to dogs residing in low-intensity housing, private homes. Serum was collected from a cross-section of 1500 adult dogs from Florida, including 500 pet dogs referred to the Veterinary Medical Center of the College of Veterinary Medicine at the University of Florida, 500 racing greyhounds, and 500 dogs residing in animal shelters. Serum was tested for D. immitis antigen, E. canis antibodies, and B. burgdorferi antibodies by ELISA. Seroprevalence of D. immitis was significantly higher (14.6%) in shelter dogs and in pet dogs (1.4%) than in racing greyhounds (0.2%) (P<0.04). There were no significant differences in the seroprevalence of E. canis (0.4-1.6%) or B. burgdorferi (0-0.8%) among the groups. There was no association of sex or age with D. immitis infection, but pit bull type dogs were more than twice as likely to be infected than other breeds (P=0.003). Evidence for vector-borne infections, particularly D. immitis, was found in dogs throughout the state. The prevalence was greatest for D. immitis infection in shelter dogs, likely due to lack of preventive medications prior to impoundment. Although heartworm infection is considered to be a treatable condition, insufficient resources in shelters may lead to euthanasia of infected dogs that would otherwise be considered adoptable.

A Diagnostic Algorithm for Detection of Antibodies to Influenza A Viruses in Dogs in Italy (2006–2008)

Since 2004, there have been several reports of Influenza A virus (FLUAV) infection in dogs. Dogs have been infected with equine influenza H3N8, avian influenza H3N2 and H5N1, and the pandemic H1N1 virus. Because ofrecent avian and equine influenza outbreaksin Italy, the objectives of the present study were to estimate the level of exposure of Italian dogs to influenza A viruses and to assess a diagnostic algorithm for detection of FLUAV exposure in dogs. Sera collected from 6,858 dogs from 2006 to 2008 were screened in a competitive enzyme-linked immunosorbent assay (cELISA) for antibodies to the highly conserved influenza A nucleoprotein. Samples positive in the cELISA were confirmed by testing in hemagglutination inhibition (HI) and fluorescent antibody test (FAT). Two seropositive dogs had antibodies to H3 hemagglutinin proteins, consistent with exposure torecent canine and equine subtype H3N8 viruses. Using a Bayesian model, the sensitivity and specificity of the cELISA were estimated as 93.98% (probability intervals [PI]: 81.67–99.08%) and 98.71% (PI: 98.43–98.96%), respectively. After accounting for the imperfect sensitivity and specificity ofthe cELISA, the Bayesian posterior prevalence of FLUAV exposure among tested Italian dogs was 0.5% (PI: 0.1–1.4%). The study results indicate that screening with a cELISA for influenza A nucleoprotein antibody, followed by confirmatory testing with HI and/or FAT, is a highly sensitive and highly specific approach for diagnosing FLUAV exposure in dogs.

Effects of anesthesia and surgery on serologic responses to vaccination in kittens

OBJECTIVE To determine the effects of anesthesia and surgery on serologic responses to vaccination in kittens. DESIGN Prospective controlled trial. ANIMALS 32 specific-pathogen-free kittens. PROCEDURES Kittens were assigned to 1 of 4 treatment groups: neutering at 7, 8, or 9 weeks of age or no neutering. All kittens were inoculated with modified-live virus vaccines against feline panleukopenia virus (FPV), feline herpesvirus (FHV), and feline calicivirus (FCV) at 8, 11, and 14 weeks of age and inactivated rabies virus (RV) at 14 weeks of age. Serum antibody titers against FPV, FHV, and FCV were determined at 8, 9, 11, 14, and 17 weeks of age; RV titers were determined at 14 and 17 weeks of age. RESULTS Serologic responses of kittens neutered at the time of first vaccination (8 weeks) were not different from those of kittens neutered 1 week before (7 weeks) or 1 week after (9 weeks) first vaccination or from those of kittens that were not neutered. In total, 31%, 0%, 69%, and 9% of kittens failed to develop adequate titers against FPV, FCV, FHV, and RV, respectively, by 17 weeks of age. CONCLUSIONS AND CLINICAL RELEVANCE Neutering at or near the time of first vaccination with a modified-live virus vaccine did not impair antibody responses in kittens. Many kittens that were last vaccinated at 14 weeks of age had inadequate antibody titers at 17 weeks of age. Kittens may be vaccinated in the perioperative period when necessary, and the primary vaccination series should be extended through at least 16 weeks of age.