Leptospira spp. in Cats in Estonia: Seroprevalence and Risk Factors for Seropositivity.

Leptospirosis is a zoonotic bacterial disease that affects humans and animals worldwide. Recently, more attention has been paid to Leptospira spp. infections in domestic cats. In this seroepidemiological study, we estimated the prevalence of anti-Leptospira spp. antibodies in domestic cats in Estonia and evaluated risk factors for the seropositivity. A total of 546 surplus feline plasma and serum samples, collected in collaboration with small animal clinics and an animal shelter in 2013 and 2015, were included in this study. The samples were tested for antibodies against Leptospira spp. using a microscopic agglutination test. The panel included Leptospira interrogans serovars Pomona, Icterohaemorrhagiae, Bratislava, Canicola, and Hardjo, and Leptospira kirschneri serovar Grippotyphosa. Titers ≥100 (positive reactions at dilutions ≥1/100) were considered positive. Anti-Leptospira spp. antibodies were detected in 12.8% of the cats. The percentage of cats that tested positive for antibodies against L. interrogans serovars Pomona, Icterohaemorrhagiae, Bratislava, Canicola, and Hardjo were 9.3%, 3.5%, 2.4%, 0.4%, and 0.2%, respectively, and the percentage of cats that tested positive for antibodies against L. kirschneri serovar Grippotyphosa was 7.3%. Of the seropositive cats, 46.5%, 35.2%, 12.7%, 4.2%, and 1.4% tested positive for 1, 2, 3, 4, and 5 serovars, respectively. The prevalence of anti-Leptospira spp. antibodies was 11.2% in pet cats and 16.3% in shelter cats. Among pet cats, the seroprevalence was over four times higher in cats that had access to the outdoors (17.2%) than in indoor cats (3.9%). Multivariable models, one based on data on pet cats only and another including also data on shelter cats, identified having access to the outdoors, being a shelter cat, and being from Western Estonia as the risk factors for seropositivity. Cats could be better protected from exposure to Leptospira spp. by not allowing them to roam freely outdoors.

Comparison of a commercial modified direct agglutination test and a commercial enzyme-linked immunosorbent assay for screening for antibodies against Toxoplasma gondii in naturally exposed domestic cats.

Domestic cats and other felids are definitive hosts for the zoonotic protozoan parasite Toxoplasma gondii. Serology is widely used in epidemiological studies conducted to estimate the proportion of domestic cats that have encountered the parasite. However, a limited number of such studies are available from some regions, including eastern parts of Europe and Russia. Various serological tests have been applied for T. gondii serology for feline samples. Seropositivity indicates previous exposure, and seropositive cats are presumed to have shed oocysts of the parasite earlier and to be chronically infected. In this study, we included a random sample of 200 sera and plasma samples from a larger sampling frame comprising samples from domestic cats from Estonia, where T. gondii is common. The samples, which had been previously screened for anti-T. gondii immunoglobulin G antibodies using a commercial modified direct agglutination test (DAT: Toxo-Screen DA; bioMérieux SA, Marcy-l'Étoile, France), were screened using a commercial enzyme-linked immunosorbent assay (ELISA: VectoToxo-antibodies [VektoTokso-antytila], VectorBest, Novosibirsk, Russian Federation). The cut-off for seropositivity with DAT was titer of 40. Of the 200 samples, 120 (60.0%) tested positive with DAT and 114 (57.0%) tested positive with ELISA; 112 samples (56.0%) tested positive with both tests. Percent agreement of 95.0% and Kappa 0.8971 indicated an almost perfect agreement between the screening results using the two methods. The results of this study can be useful for comparison, evaluation, and interpretation of results obtained with these two tests in seroepidemiological studies and may encourage more studies on the topic from eastern parts of Europe and Russia.

Toxoplasma gondii Seroprevalence in Horses from Ukraine: an Investigation Using Two Serological Methods.

BACKGROUND: Horses are recognized as important hosts for the zoonotic parasite Toxoplasma gondii, mainly because meat of infected horses can be a source of human T. gondii infections. However, the baseline knowledge on equine T. gondii infections is limited and lacking from many countries. This seroepidemiological study was set in Ukraine, a country where little is known about T. gondii prevalence in any host species. METHODS: We investigated 78 serum samples from horses, collected from Kyiv and Lviv regions of Ukraine, using a commercial enzyme-linked immunosorbent assay (ELISA) and a commercial modified direct agglutination test (DAT). The ELISA is intended to detect the presence of total antibodies against T. gondii, and the DAT detects specific anti-T. gondii immunoglobulin G antibodies. The cutoff titer for DAT seropositivity was a titer of 40. RESULTS: Two horses were omitted from statistical analyses due to disagreeing ELISA results. Altogether 16 of the remaining 76 horses were seropositive with the ELISA, yielding an apparent seroprevalence estimate of 21.1% (95% confidence interval 13.0-31.3). Five horses tested positive with both ELISA and DAT, 11 horses tested positive with ELISA only, and one with DAT only. For 64 of the 76 horses, the results of ELISA and DAT agreed; the percent agreement was 84.2 and the Kappa statistic was - 1.5208, indicating poor agreement. CONCLUSION: Our results add to the knowledge on the global epidemiology of T. gondii and exposure level of horses to the major zoonotic parasite, as well as to the discussion of challenges of applying serology to estimate exposure to T. gondii in horses.

Seroprevalence of Toxoplasma gondii in domestic pigs, sheep, cattle, wild boars, and moose in the Nordic-Baltic region: A systematic review and meta-analysis.

BACKGROUND: Toxoplasma gondii is an important foodborne zoonotic parasite. Meat of infected animals is presumed to constitute a major source of human infection and may be a driver of geographical variation in the prevalence of anti-T. gondii antibodies in humans, which is substantial in the Nordic-Baltic region in northern Europe. However, data on seroprevalence of T. gondii in different animal species used for human consumption are scattered. METHODS: We conducted a systematic review of seroprevalence studies and meta-analysis to estimate the seroprevalence of T. gondii in five animal species that are raised or hunted for human consumption in the Nordic-Baltic region: domestic pigs (Sus scrofa domesticus), sheep (Ovis aries), cattle (Bos taurus), wild boars (Sus scrofa), and moose (Alces alces). We searched for studies that were conducted between January 1990 and June 2018, and reported in articles, theses, conference abstracts and proceedings, and manuscripts. Subgroup analyses were performed to identify variables influencing the seroprevalence. FINDINGS: From a total of 271 studies identified in the systematic review, 32 were included in the meta-analysis. These comprised of 13 studies on domestic pigs, six on sheep, three on cattle, six on wild boars, and four on moose. The estimated pooled seroprevalence of T. gondii was 6% in domestic pigs (CI95%: 3-10%), 23% in sheep (CI95%: 12-36%), 7% in cattle (CI95%: 1-21%), 33% in wild boars (CI95%: 26-41%), and 16% in moose (CI95%: 10-23%). High heterogeneity was observed in the seroprevalence data within each species. In all host species except wild boars, the pooled seroprevalence estimates were significantly higher in animals >1 year of age than in younger animals. Not all studies provided information on animal age, sensitivity and specificity of the serological method employed, and the cut-off values used for defining an animal seropositive. CONCLUSIONS: A substantial proportion of animals raised or hunted for human consumption in the region had tested positive for T. gondii. This indicates widespread exposure to T. gondii among animals raised or hunted for human consumption in the region. Large variations were observed in the seroprevalence estimates between the studies in the region; however, studies were too few to identify spatial patterns at country-level.

Toxoplasma gondii seroprevalence in free-ranging moose (Alces alces) hunted for human consumption in Estonia: Indicator host species for environmental Toxoplasma gondii oocyst contamination.

In Estonia, northeastern Europe, antibodies against Toxoplasma gondii are common in many host species, including wildlife, domestic animals, and humans. Our nationwide study aimed to estimate T. gondii seroprevalence and its geographical distribution, and to evaluate plausible risk factors for seropositivity in free-ranging moose (Alces alces) hunted for human consumption in 2015. We screened sera or plasma from 463 moose for presence of anti-T. gondii immunoglobulin G antibodies with a commercial direct agglutination test, using a cut-off titer 40 for seropositivity. Altogether 111 moose tested seropositive, yielding a seroprevalence estimate of 23.97%. Seropositive moose were detected in all the 13 counties where the samples originated from. Based on a multivariable logistic regression model, 'being female', 'being adult', and 'being from the northern part of the country' were significant risk factors, with odds ratios of 2.91, 3.07, and 3.11, respectively, and there was interaction between the variables 'being female' and 'being from the northern part of the country'. A substantial proportion of the moose investigated had been exposed to T. gondii. Presence of the parasite in edible tissues of the moose was not shown in the present study, but moose hunted in Estonia should be considered a potential T. gondii infection source to other hosts, including humans. Seropositivity indicates previous exposure, and because the seroprevalence was higher in adults than in calves and because moose are herbivores, the exposure was likely exposure to and ingestion of T. gondii oocysts. The results can thus be interpreted to indicate that the environment in Estonia was widely contaminated with T. gondii oocysts, in particular in the northern part of the country. Investigation of samples from a free-ranging herbivorous host that is hunted in large numbers appears useful in revealing environmental distribution patterns of T. gondii.

Toxoplasma gondii seroprevalence in breeding pigs in Estonia.

BACKGROUND: Toxoplasma gondii is a widespread occurring parasite infecting warm-blooded animals, including pigs and humans. The aims of this study were to estimate the prevalence of anti-T. gondii antibodies and to evaluate risk factors for T. gondii seropositivity in breeding pigs raised in Estonia. Sera from 382 pigs were tested with a commercial direct agglutination test, using a cut-off titer of 40 for seropositivity, for the presence of anti-T. gondii immunoglobulin G antibodies. RESULTS: Twenty-two (5.8%) of the 382 pigs tested seropositive for T. gondii, and 6 of the 14 herds had at least one seropositive pig. The proportion of seropositive pigs within the herds ranged between 0 and 43%. Gender appeared as a significant factor, with sows having 5.6 times higher odds to be seropositive to T. gondii than boars. Seroprevalence did not increase with age. CONCLUSIONS: Anti-T. gondii antibodies were present in a substantial proportion of breeding pig herds in Estonia. On the other hand, the presence of herds without seropositive pigs illustrates that porcine T. gondii infections can be avoided even in a country where the parasite is endemic and common in several other host species.

Toxoplasma gondii seroprevalence varies by cat breed.

Toxoplasma gondii is a widespread zoonotic parasite that is relevant for veterinary and public health. The domestic cat, the definitive host species with the largest worldwide population, has become evolutionarily and epidemiologically the most important host of T. gondii. The outcome of T. gondii infection is influenced by congenital and acquired host characteristics. We detected differences in T. gondii seroprevalence by cat breed in our previous studies. The aims of this study were to estimate T. gondii seroprevalence in selected domestic cat breeds, and to evaluate whether being of a certain breed is associated with T. gondii seropositivity, when the age and lifestyle of the cat are taken into account. The studied breeds were the Birman, British Shorthair, Burmese, Korat, Norwegian Forest Cat, Ocicat, Persian, and Siamese. Plasma samples were analyzed for the presence of immunoglobulin G antibodies against T. gondii with a commercial direct agglutination test at dilution 1:40. The samples were accompanied by owner-completed questionnaires that provided background data on the cats. Overall, 41.12% of the 1121 cats tested seropositive, and the seroprevalence increased with age. The Burmese had the lowest seroprevalence (18.82%) and the Persian had the highest (60.00%). According to the final multivariable logistic regression model, the odds to test seropositive were four to seven times higher in Birmans, Ocicats, Norwegian Forest Cats, and Persians when compared with the Burmese, while older age and receiving raw meat were also risk factors for T. gondii seropositivity. This study showed that T. gondii seroprevalence varies by cat breed and identified being of certain breeds, older age, and receiving raw meat as risk factors for seropositivity.

Seroprevalence of and Risk Factors for Toxoplasma gondii Infection in Cats in Estonia.

In Estonia, northeastern Europe, Toxoplasma gondii seroprevalence in humans has not declined, in contrast to many other countries. The reasons for this are unknown. Domestic cats are important hosts in the epidemiology of the parasite, but information on local feline T. gondii infections has been lacking. An epidemiological cross-sectional study was conducted to estimate the seroprevalence of T. gondii and the risk factors associated with seropositivity in cats in Estonia. Surplus from blood samples that had been collected for unrelated diagnostic purposes from 306 pet cats and 184 shelter cats were analyzed for anti-T. gondii immunoglobulin G antibodies by using a direct agglutination test. Two questionnaires were designed to reveal relevant risk factors for seropositivity. The overall seroprevalence of T. gondii in cats in Estonia was 60.8%. Older age, outdoor access, hunting, living outside the city in the countryside, and not being a purebred cat were among the risk factors associated with seropositivity. T. gondii is highly prevalent in domestic cats in Estonia. This suggests that the environment has been contaminated with T. gondii. Seropositivity indicates previous oocyst shedding, and most of the cats had outdoor access. The increase in T. gondii seroprevalence with age indicates acquired infections, and most of the risk factors were lifestyle-related. Cat owners could diminish the risk of T. gondii infection and also limit the spread of the parasite by not allowing their cats to roam free.