Toxoplasma gondii is one of the most prevalent zoonotic protozoan parasites worldwide and affects the vast majority of warm-blooded animal species, including humans. Postnatal infection in humans occurs through the ingestion of sporulated T. gondii oocysts or via the oral intake of parasite tissue cysts during the consumption of raw or undercooked meat. In this regard, given their high exposure to oocysts, chickens (Gallus domesticus) raised on the ground constitute a potential source of T. gondii. For the first time in Spain, a survey was undertaken in commercial retail free-range poultry. A total of 50 thighs from different animals were analysed. The samples were homogenised and an acid pepsin digestion procedure was applied prior to molecular analysis. Toxoplasma gondii DNA was isolated from meat by qPCR. Two sets of primers were used for DNA amplification targeting the specific sequence of a 529 bp repeat element and another set of primers was utilised for the surface antigen protein-1 gene. DNA extracted from 5 out of 50 tissue samples was positive for both genes by qPCR amplification. The 10% prevalence of Toxoplasma infection found in commercial free-range chickens raises public health issues.

Four-week-old chickens were inoculated orally with 1,000 or 100,000 oocysts of the ME-49 or GT-1 strain of Toxoplasma gondii, and their antibody responses were measured, using the direct modified agglutination test, latex agglutination test, indirect hemagglutination test, ELISA, and the Sabin-Feldman dye test. Antibodies against T gondii were detected by use of the modified agglutination test and ELISA within 2 weeks of oocyst inoculation, and antibodies persisted until termination of the study by postinoculation day 68. The latex agglutination test was insensitive in detecting T gondii antibodies, and antibodies were not detected by use of the dye and indirect hemagglutination tests. Of tissues bioassayed in mice for tissue cysts by pepsin digestion of individual organs of chickens on postinoculation day 68, tissue cysts were found in the brain of all 5, heart of 3, and leg muscles of 2, but not in the liver and breast muscles. None of the birds developed clinical toxoplasmosis.

Kittens are the principal disseminators of Toxoplasma gondii. They can shed > 10(8) oocysts in the feces after initial infection with bradyzoites in tissue cysts. Thereafter, most kittens develop protective immunity and do not shed oocysts again if they are reinfected. Bradyzoites of a T gondii mutant, designated T-263, were used to vaccinate kittens. Their use did not result in oocyst shedding, but successfully prevented 84% (31/37) of the kittens from shedding oocysts when challenge exposed with a normal isolate of T gondii. Vaccination of outdoor-roaming cats and kittens would be a useful public health measure to prevent transmission of toxoplasmosis near homes, on farms, and in zoos. It is anticipated that several years will be required for a lyophilized bradyzoite vaccine to be ready for licensing and possible commercial availability.

OBJECTIVE To determine the extent of environmental exposure to heteroxenous coccidia from wild canid feces in southeastern Ohio. SAMPLE 285 presumed wild canid fecal samples collected across an ecological system in southeastern Ohio. PROCEDURES Morphological classification and molecular analysis were used to determine the canid genus for collected fecal samples. Microscopic and molecular analysis were used to detect coccidian oocysts and DNA. Several variables were analyzed for associations with coccidian DNA detection or prevalence. RESULTS Coccidian DNA was detected in 51 of 285 (17.9%) fecal samples. Of those positive samples, 1% (95% confidence interval, 0.4% to 3%) had positive results for Hammondia heydorni and none had positive results for Neospora caninum, for an estimated environmental N caninum prevalence of 0% (95% confidence interval, 0% to 7%)/1-km2 hexagonal area evaluated. Morphological classification revealed that 78.9% (225/285) of fecal samples were from coyotes and 17.2% (49/285) were from foxes. No difference in proportions of coccidian DNA-positive fecal samples was identified among canid species. Environmental temperature and fecal freshness were associated with coccidian DNA detection. Land use type, relative canid density, and cattle density were not associated with the prevalence of coccidian DNA-positive samples. CONCLUSIONS AND CLINICAL RELEVANCE The low prevalence of coccidia shed in wild canid feces in this study, including the estimated 0% environmental prevalence of N caninum, suggested that the role of the oocyst environmental phase in coccidia transmission to ruminants is likely minor in rural southeastern Ohio.

During the United States Department of Agriculture (USDA) National Animal Health Monitoring System’s (NAHMS) 2007‐2008 beef study, 567 producers from 24 US States were offered the opportunity to collect fecal samples from weaned beef calves and have them evaluated for the presence of parasite eggs (Phase 1). Participating producers were provided with instructions and materials for sample collection. Up to 20 fresh fecal samples were collected from each of the 99 participating operations. Fresh fecal samples were submitted to one of 3 randomly assigned laboratories for evaluation. Upon arrival at the laboratories, all samples were processed for the enumeration of strongyle, Nematodirus, and Trichuris eggs using the modified Wisconsin technique. The presence or absence of coccidian oocysts and tapeworm eggs was also noted. In submissions where the strongyle eggs per gram exceeded 30, aliquots from 2 to 6 animals were pooled for DNA extraction. Extracted DNA was subjected to genus level polymerase chain reaction (PCR) identification for the presence of Ostertagia, Cooperia, Haemonchus, Oesophagostomum, and Trichostrongylus. In this study, 85.6% of the samples had strongyle type, Nematodirus, and Trichuris eggs. Among the samples evaluated, 91% had Cooperia, 79% Ostertagia, 53% Haemonchus, 38% Oesophagostomum, 18% Nematodirus, 7% Trichuris, and 3% Trichostrongylus. The prevalence of coccidia and tapeworm eggs was 59.9% and 13.7%, respectively.

During the United States Department of Agriculture (USDA) National Animal Health Monitoring System’s (NAHMS) 2007‐2008 beef study, producers from 24 states were offered the opportunity to evaluate their animals for internal parasites and for overall responses to treatment with anthelmintics. A lapse of 45 d was required between initial sampling and any previous treatments. Choice of anthelmintic (oral benzimidazoles, and both injectable and pour-on endectocides) was at the discretion of the producer so as not to alter the local control programs. Fresh fecal samples were collected from 20 animals, or from the entire group if less than 20, then randomly assigned to 1 of 3 participating laboratories for examination. Analyses consisted of double centrifugation flotation followed by enumeration of strongyle, Nematodirus, and Trichuris eggs (the presence of coccidian oocysts and tapeworm eggs was also noted). Where strongyle eggs per gram (epg) exceeded 30, aliquots from 2 to 6 animals were pooled for egg isolation and polymerase chain reaction (PCR) analysis for the presence of Ostertagia, Cooperia, Haemonchus, Oesophagostomum, and Trichostrongylus. Results from 72 producers (19 States) indicated that fecal egg count reductions were < 90% in 1/3 of the operations. All operations exhibiting less than a 90% reduction had used pour-on macrocyclic lactones as the anthelmintic treatment. While some of these less than expected reductions could have been the result of improper drug application, PCR analyses of the parasite populations surviving treatment, coupled with follow-up studies at a limited number of sites, indicated that less than expected reductions were most likely due to anthelmintic resistance in Cooperia spp. and possibly Haemonchus spp.

Proteins in sporulated and unsporulated oocysts of Eimeria maxima were characterized, using monoclonal antibodies (MAB), ELISA, sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), and protein (western) immunoblotting techniques. Three MAB (EM1, EM2, and EM4) were produced against proteins of sporulated oocysts. The ELISA results indicated that EM1 was reactive with sporulated oocyst proteins, EM2 was reactive with sporulated and unsporulated oocyst proteins, and EM4 was reactive with unsporulated oocysts and proteins. Separation of proteins in E maxima sporulated and unsporulated oocysts by SDS-PAGE indicated that sporulated oocysts had proteins of approximately 200 kilodaltons (kD) and distinct protein bands at 21.5 and 45 kD. Using SDS-PAGE, unsporulated oocysts had less-distinct high molecular weight protein bands (> 200 kD), compared with sporulated oocysts, and a distinct protein band at 31 kD. Use of all 3 MAB yielded negative results in western blot analysis of fractions obtained by SDS-PAGE.

OBJECTIVE To evaluate whether anti-inflammatory doses of cyclosporine activate Toxoplasma gondii in chronically infected cats or potentiate infection in cats exposed for the first time. ANIMALS 30 T gondii–negative cats. PROCEDURES Cats were assigned to 1 of 3 groups (10 cats/group). Group 1 (control) cats were administered a placebo for 126 days; group 2 cats were administered a placebo for 84 days, followed by cyclosporine at 7.5 mg/kg/d, PO, for 42 days; and group 3 cats were administered cyclosporine at 7.5 mg/kg/d, PO, for 126 days. Cats were orally inoculated with T gondii on day 42. Results for fecal flotations, PCR assays, and histologic examinations and IgM and IgG titers were analyzed. Cyclosporine concentrations were measured on selected days. RESULTS All cats were infected by T gondii and developed signs of self-limiting gastrointestinal tract infection. Group 3 had the highest incidence and severity of CNS and pulmonary histopathologic findings typical of toxoplasmosis. One cat in group 3 died of systemic toxoplasmosis; that cat had a cyclosporine concentration of 1,690 ng/mL. Group 2 cats infected with T gondii before cyclosporine administration did not have repeated oocyst shedding. Group 3 cats shed fewer oocysts for a shorter time than did control cats of group 1. CONCLUSIONS AND CLINICAL RELEVANCE Oral administration of cyclosporine in accordance with the protocol for this study did not potentiate the enteroepithelial phase of T gondii infection. Cats with high cyclosporine blood concentrations at the time of primary T gondii infection may be at risk of developing systemic toxoplasmosis.

Four 1-year-old steers were each inoculated orally with 10,000 Toxoplasma gondii oocysts of the GT-1 strain and euthanatized on postinoculation days (PID) 350, 539, 1191, and 1201. Samples (500 g) of tongue, heart, semimembranosus and semitendinosus muscles (roast), intercostal muscles (ribs), longismus muscles (tenderloin), brain, kidneys, liver, and small intestine were bioassayed for T. gondii by feeding to cats and examination of cat feces for shedding of oocysts. Toxoplasma gondii was recovered by bioassays in cats from the 3 steers necropsied PID 350, 539, and 1191, but not from the steer euthanatized on PID 1201. Cats shed oocysts after ingesting tongue from 2 steers, heart from 3 steers, liver from 2 steers, and roast, ribs, brain, and intestines from 1 steer each. Taxoplasma gondii was not isolated from any of the other bovine tissues. In addition to tissues bioassayed in cats, homogenates of mesenteric lymph nodes, lungs, spinal cord, spleen, and eyes were bioassayed in mice for T. gondii infection. Toxoplasma gondii was not recovered from the 135 mice inoculated with tissue from each of the 4 steers. All 4 inoculated steers developed high T. gondii antibody titers (greater than or equal to 1:8,000) in the agglutination test, using formalin-fixed whole tachyzoites. In the steer euthanatized on PID 1201, agglutinating T. gondii antibody titers decreased from 1:4,000 to 1:320 between 2 and 5 months after inoculation and to 1:20 by 19 months after inoculation.