Association of Wolbachia with heartworm disease in cats and dogs.

Although the presence of adult Dirofilaria immitis in the pulmonary arteries and its associated arteritis and thromboembolic disease can explain some of the manifestations of canine and feline heartworm disease, the cause of other findings remains unclear. Cats with D. immitis antibodies but lacking adult parasites in the pulmonary arteries frequently develop histological lesions of the airways, resulting in a condition termed Heartworm-Associated Respiratory Disease. All D. immitis parasites harbor Wolbachia pipientis bacteria and D. immitis-infected animals can have circulating Wolbachia antibodies and pro-inflammatory Wolbachia antigens (WSP) deposited in tissues. Little is known about the role that Wolbachia plays in lung disease of animals naturally infected with D. immitis. The purpose of this study was to determine the contribution of Wolbachia to the pathogenesis of natural heartworm disease in cats and dogs. We hypothesized that animals having sufficient Wolbachia burden to be detected in lung tissue by immunohistochemistry and/or polymerase chain reaction (PCR) would have more severe pulmonary disease than those with bacteria below the limits of detection. We further hypothesized that animals that were immunoreactive to pro-inflammatory WSP would have more severe pulmonary lesions than those that were seronegative for WSP antibodies. Blood and lung tissue samples were collected from cats and dogs representing three different D. immitis infection statuses: heartworm-free, heartworm-exposed, heartworm-infected. There was a positive but weak correlation between the magnitude of D. immitis antibody titers and WSP titers in cats (r=0.57, p<0.001) and in dogs (r=0.39, p<0.001). Pulmonary lesions were more common in HW-infected animals than in HW-free animals. Pulmonary arteriolar occlusion was more common in HW-infected cats (57%; p=0.003) than in HW-infected dogs (17%). Although pulmonary lesions were most common in HW-infected animals, there was no clear additive effect when either Wolbachia DNA/WSP was detected in lung tissue or when circulating Wolbachia antibodies were detected. There were no significant differences in the magnitude of pulmonary lesion scores within each HW-infection status group regardless of whether Wolbachia DNA/WSP or antibodies were detected. The relationship between Wolbachia and lung pathology in heartworm-infected animals remains to be determined. The lack of clear evidence for a role of Wolbachia in heartworm disease creates a dilemma for veterinarians treating animals in D. immitis-endemic areas. Although the indiscriminant use of antibiotics should be avoided, many clinicians prescribe doxycycline based on the favorable responses observed in human filarial diseases and promising results from the first published studies of doxycycline use in D. immitis-infected dogs.

Relationship between separation time of plasma from heparinized whole blood on plasma biochemical analytes of loggerhead sea turtles (Caretta caretta).

Concentrations and activities of selected biochemicals of loggerhead sea turtles (Caretta caretta) were determined for plasma that was separated from whole blood samples that were kept up to 96 hr post collection (PC) in a refrigerator. Blood samples collected from seven juvenile captive loggerhead sea turtles were added to tubes containing lithium heparin and were placed on ice. Equal amounts of anticoagulated whole blood from the lithium heparin tubes were then aliquoted into plastic tubes and stored as whole blood under refrigeration until they were centrifuged at 0, 4, 24, 48, and 96 hr PC. Plasma was removed and the analytes that were measured were alkaline phosphatase (ALP), aspartate aminotransferase (AST), gamma glutamyl transferase (GGT), creatine kinase (CK), sodium, chloride, potassium, magnesium, calcium, phosphorus, cholesterol, glucose, urea nitrogen, uric acid, total protein, albumin, and globulin. Compared with values at 0 time, the only analyte to be significantly different at 24 hr PC was GGT (activity decreased by 25%). Compared with values at 0 time, significant differences at 96 hr PC were only seen in AST (2% increase), GGT (25% decrease), glucose (7% decrease), and uric acid (25% increase). Although a statistically significant difference was found in concentrations of phosphorus and cholesterol over time by repeated measures analysis of variance (ANOVA), the follow-up multiple comparison procedure could not define the specific time points at which significant differences occurred. For all other analytes, significant differences over the time course of the study were not found. In these instances, the power of the ANOVA was sufficient (> or = 0.80) to detect any arithmetic differences of a clinically relevant magnitude. Although plasma should be separated from the cellular component of blood as soon as possible PC, in a field situation in which a centrifuge is unavailable, samples can be stored in a portable cooler up to 24 hr without appreciable change in select biochemical analytes.

Prevalence of anthelmintic resistant cyathostomes on horse farms.

OBJECTIVE: To determine prevalence of anthelmintic resistance in cyathostome nematodes of horses in the southern United States. DESIGN: Cross-sectional study. ANIMALS: 786 horses on 44 farms and stables in Georgia, South Carolina, Florida, Kentucky, and Louisiana. PROCEDURE: Fecal egg count (FEC) reduction tests were performed on 44 large farms and stables. Horses on each farm were treated with an oral paste formulation of fenbendazole, oxibendazole, pyrantel pamoate, or ivermectin at recommended label dosages. A mixed linear model was fitted to the percentage reduction in FEC, accounting for differences among farms, states, ages, treatments, and treatment by state interactions. RESULTS: By use of a conservative measure of resistance (< 80% reduction), the percentage of farms with anthelmintic-resistant cyathostomes was 97.7%, 0%, 53.5%, and 40.5% for fenbendazole, ivermectin, oxibendazole, and pyrantel pamoate, respectively. Mean percentage reductions in FEC for all farms were 24.8%, 99.9%, 73.8%, and 78.6% for fenbendazole, ivermectin, oxibendazole, and pyrantel pamoate, respectively. Pairwise contrasts between states for each treatment revealed that in almost all instances, there were no significant differences in results between states. CONCLUSIONS AND CLINICAL RELEVANCE: The prevalence of resistance found in this study was higher than that reported previously, suggesting that anthelmintic resistance in equine cyathostomes is becoming a major problem. Furthermore, data from these 5 southern states, which are geographically and physiographically distinct, were remarkably similar. This suggests that drug resistance in cyathostomes is highly prevalent throughout the entire southern United States and probably nationwide.

UNUSUAL POPULATION GENETICS OF A PARASITIC NEMATODE: mtDNA VARIATION WITHIN AND AMONG POPULATIONS.

Very little is known about the distribution of genetic variance within and among populations of parasitic helminths. In this study we used mitochondrial DNA (mtDNA) restriction fragment analysis to describe the population genetic structure of Ostertagia ostertagi, a nematode parasite of cattle, in the United States. Estimates of within-population mtDNA diversity are 5 to 10 times greater than typical estimates reported for species in other taxa. Although populations are genetically differentiated for a key life-history trait, greater than 98% of the total genetic diversity is partitioned within populations, and the geographic distribution of individual mtDNA haplotypes suggests high gene flow among populations.