Seroprevalence and risk factors of Toxoplasma gondii infection in goats from South Punjab Province, Pakistan.

Layyah District in South Punjab Province of Pakistan offers the most intensive caprine economy in the country; its Indus riverine and desert environment makes the area peculiar and worthy of specific investigations. This study aimed to investigate the prevalence of anti-Toxoplasma gondii (T. gondii) IgG-antibody in goats in serum samples and the potential risk factors. The prevalence of T. gondii infection was estimated using a two-stage sample design. All caprine farms in the study area were stratified by size, and from these 110 were randomly selected. Twelve goats (>1-year-old) were selected from each farm and a total of 1320 serum samples were collected and tested by ELISA. A questionnaire on the conditions and management practices of each farm was administered to 110 farmers. Four hundred and sixteen out of 1320 sera samples (31.5%) were found positive and 89% of the flock had at least one seropositive goat. The proportion of seropositive goats tested within each flock ranged from 8.3% to 83.3%. with several factors contributing to this heterogeneity. Goat age played a significant role in the presence of cats. Significant interactions were related to goat farms having floor of dirt and kitten presence. Moreover, age class, abortion history and water source supply were modulated by owner education levels. This is the first study to determine the prevalence of anti-T. gondii antibodies in goats sera in Layyah district and the largest carried out so far in Pakistan. The remarkable presence of T. gondii among goats in areas where goat farming plays a significant economic role may pose a production threat to the small-stock industry, as well as to public health and food safety. Therefore, investigations to identify high-risk goat populations are highly recommended in order to facilitate the implementation of local control strategies.

Molecular detection of Toxoplasma gondii and Neospora caninum in seabirds collected along the coast of Santa Catarina, Brazil.

Toxoplasma gondii and Neospora caninum are two closely related protozoans that infect a wide range of animals, including birds. However, the occurrence of N. caninum and T. gondii in seabirds is unknown. Therefore, this study aimed to determine the presence of T. gondii and N. caninum DNA in tissue samples of seabirds. Tissue samples of the pectoral muscles, heart, and brain were collected from 47 birds along the coastline of Santa Catarina State, SC, Brazil. The DNA was extracted from the tissues and screened using nested-PCR (nPCR) targeting internal transcribed spacer 1 (ITS1). T. gondii DNA was detected in tissues from seven seabirds (7/47, 14.8%), kelp gull (Larus dominicanus) (5/21), and Manx shearwater (Puffinus puffinus) (2/8). N. caninum DNA was detected in tissues of nine seabirds (9/47, 19.1%), the kelp gull (L. dominicanus) (4/21), Manx shearwater (P. puffinus) (2/8), neotropic cormorant (Phalacrocorax brasilianus) (1/4), brown booby (Sula leucogaster) (1/5), and white-chinned petrel (Procellaria aequinoctialis) (1/1); however, no co-infection was observed. In conclusion, this study showed the circulation of N. caninum and T. gondii in seabirds along the coastline of Santa Catarina State. Further studies are required to clarify the role of these birds in the epidemiology of neosporosis and toxoplasmosis.

Seroprevalence and risk factors of Toxoplasma gondii in sheep and goats of North West Province, South Africa.

BACKGROUND: The protozoan parasite Toxoplasma gondii causes toxoplasmosis, one of the most prevalent parasitic zoonotic diseases with significant economic and public health implications worldwide. Infection with the parasite has a significant adverse effect on sheep and goat production and can frequently go undetected in the herd, resulting in abortions and weak or dead offspring. Although there are few studies on seroprevalence and risk factors associated with T. gondii infections in livestock in other provinces of South Africa, there is no data in the North West province. Therefore, a cross-sectional study was conducted to investigate the seroprevalence of T. gondii and risk factors associated with exposure in sheep and goats of the North West province of South Africa. Sera from 439 livestock (164 sheep and 285 goats) were collected and analysed for the presence of T. gondii IgG antibodies using indirect ELISA (Enzyme-linked immunosorbent assay). An assessment of potential risk factors in farms associated with seropositivity was also conducted using a structured questionnaire. RESULTS: Out of the 439 tested sheep and goats, 13.9% (61/439) were positive for IgG antibodies against T. gondii. Sheep and goats had seroprevalences of 19.5% (32/164) and 10.5% (29/275) respectively. In the multivariable logistic regression model, the risk of acquiring T. gondii was significantly higher in the mixed breed [Odds ratio (OR) = 71.07; 95% confidence interval (CI): 266.8-1893.1; p < 0.011)] animals than white dorper sheep and in farms that burn or bury aborted material (OR = 42.04; CI: 179.9-982.5; p = 0.020) compared to those that only burn aborted material. The risk was lower for the farms in Kagisano-Molopo (OR = 0.00; CI: 0.0-25.4; p = 0.015) and Mahikeng (OR = 0.00; CI: 0.0-4.9; p < 0.001) local municipalities than Greater Taung local municipality, and for the animals that drink water from dams (OR = 0.03; CI: 0.2-58.8; p = 0.021) than those that drink from boreholes. CONCLUSION: The seroprevalence and risk factors associated with transmission observed show that T. gondii infection is widespread in sheep and goats of the North West province.

Observational longitudinal study on Toxoplasma gondii infection in fattening beef cattle: serology and associated haematological findings.

Toxoplasmosis, caused by the protozoan parasite Toxoplasma gondii, is a globally distributed zoonotic infection with significant implications for human and animal health. This study investigated the prevalence of T. gondii infection in a population of beef cattle at three different stages of their productive lifespan and examined the impact of T. gondii serological status on blood parameters. A commercial beef fattening unit in Italy was the setting for this research, which involved a biosecurity assessment upon cattle arrival, blood sampling at three time points and Toxoplasma-specific serological testing using indirect fluorescent antibody tests (IFAT). Results revealed a dynamic pattern of T. gondii seropositivity in cattle, with an initial prevalence of 30.6% at arrival (T0) that increased to 44.6% at 14 days (T1) and then decreased slightly to 39.3% at slaughter after 5 months (T2). Interestingly, seroconversion was observed during the study, indicating ongoing infections, and antibody waning occurred in some animals. In terms of blood parameters, seropositive cattle exhibited significantly lower mean corpuscular volume (MCV) and a higher neutrophil-lymphocyte (N/L) ratio, suggesting an activation of the innate immune response. Furthermore, cattle with higher antibody titres displayed higher neutrophil counts. However, all blood parameters with a statistical significance were within the reference range. This study provides for the first time a longitudinal investigation on the serological status for T. gondii in naturally exposed beef cattle. These findings provide valuable insights into the clinico-pathological aspects of natural T. gondii exposure in cattle and underscore the importance of monitoring and managing T. gondii infection in livestock production systems.

Prevalence and factors associated with Leishmania spp. and Toxoplasma gondii infections in apparently healthy horses in Eastern Spain.

Leishmaniasis and toxoplasmosis are two of the most common parasitic zoonoses. Leishmaniasis is endemic to 98 countries around the world, whereas toxoplasmosis is widely distributed throughout the world, causing significant health expenditure. Horses can play a relevant role in the transmission of the disease, being a silent reservoir, as clinical signs are not common. Serum samples from 166 horses living in eastern Spain (Mediterranean basin) were analysed to determine the presence of antibodies against Leishmania spp. and T. gondii by ELISA (Enzyme-linked Immunosorbent Assay.) The risk factors evaluated were the geographical area and the relative humidity and average temperature, and epidemiological factors such as sex, reproductive status, age, breed, morphotype, living with other domestic animals, use and access to the outdoors. Seroprevalence of Leishmania spp. and T. gondii infection was found 28.92%, and 16.27% respectively, whereas co-infection of the two parasites was found only in two males. Leishmania seroprevalence was high in castrated males and several mesodolichomorphic equine breeds used for teaching, as well as in outdoor animals. The most elevated seroprevalence was found in winter with higher levels of rainfall, whereas high seroprevalence of T. gondii was found in crossbreeding animals and those used for breeding. High seroprevalence of Leishmania spp. and T. gondii was found in horses of the Mediterranean basin. These data suggest that horses can act as a silent reservoir and that this species has high potential for transmission to humans, outdoor animals and in geographical areas with high average rainfall.

In vitro assay to determine inactivation of Toxoplasma gondii in meat samples.

Consumption of raw and undercooked meat is considered as an important source of Toxoplasma gondii infections. However, most non-heated meat products contain salt and additives, which affect T. gondii viability. It was our aim to develop an in vitro method to substitute the mouse bioassay for determining the effect of salting on T. gondii viability. Two sheep were experimentally infected by oral inoculation with 6.5 × 104 oocysts. Grinded meat samples of 50 g were prepared from heart, diaphragm, and four meat cuts. Also, pooled meat samples were either kept untreated (positive control), frozen (negative control) or supplemented with 0.6 %, 0.9 %, 1.2 % or 2.7 % NaCl. All samples were digested in pepsin-HCl solution, and digests were inoculated in duplicate onto monolayers of RK13 (a rabbit kidney cell line). Cells were maintained for up to four weeks and parasite growth was monitored by assessing Cq-values using the T. gondii qPCR on cell culture supernatant in intervals of one week and ΔCq-values determined. Additionally, 500 µL of each digest from the individual meat cuts, heart and diaphragm were inoculated in duplicate in IFNγ KO mice. Both sheep developed an antibody response and tissue samples contained similar concentrations of T. gondii DNA. From all untreated meat samples positive ΔCq-values were obtained in the in vitro assay, indicating presence and multiplication of viable parasites. This was in line with the mouse bioassay, with the exception of a negative mouse bioassay on one heart sample. Samples supplemented with 0.6 %-1.2 % NaCl showed positive ΔCq-values over time. The frozen sample and the sample supplemented with 2.7 % NaCl remained qPCR positive but with high Cq-values, which indicated no growth. In conclusion, the in vitro method has successfully been used to detect viable T. gondii in tissues of experimentally infected sheep, and a clear difference in T. gondii viability was observed between the samples supplemented with 2.7 % NaCl and those with 1.2 % NaCl or less.

Genotyping of Toxoplasma gondii in domestic animals from Campeche, México, reveals virulent genotypes and a recombinant ROP5 allele.

Toxoplasma gondii has at least 318 genotypes distributed worldwide, and tropical regions usually have greater genetic diversity. Campeche is a state located in the southeastern region of México and has favourable climate conditions for the replication and dissemination of this protozoan, similar to those in South American countries where broad genetic diversity has been described. Thus, in this study, 4 T. gondii isolates were obtained from tissues of stray dogs and free-range chickens in Campeche, México, and were genotyped by Mn-PCR-RFLP with 10 typing markers (SAG1, altSAG2, SAG3, BTUB, GRA6, c22-8, c29-2, L358, PK1 and Apico) and 5 virulence markers (CS3, ROP16, ROP17, ROP18 and ROP5) to provide new information about the distribution and virulence prediction of T. gondii genotypes. Two isolates of T. gondii genotype #116 and 2 of genotype #38 were obtained from stray dogs and chickens, respectively. The parasite load found in these species was between <50 and more than 35 000 tachyzoites per mg of tissue. Virulence marker genotyping revealed a recombinant 1&3 ROP5 RFLP pattern in 2 ToxoDB #116 isolates with no prediction of virulence in a murine model, while in the 2 ToxoDB #38 isolates, the ROP18/ROP5 combination predicted high virulence. Considering all the typed markers, there is a predominance of type I and III alleles, as constantly reported for the isolates characterized in various regions of México. It is crucial to determine their phenotype to corroborate the genetic virulence profile of the T. gondii isolates obtained in this study.

Seroepidemological investigation of Toxoplasma gondii and Trichinella spp. in pigs reared by tribal communities and small-holder livestock farmers in Northeastern India.

Toxoplasma gondii and Trichinella spp. are critical tissue-dwelling foodborne zoonotic parasites associated with pork consumption and pig rearing. Despite being a major pig-rearing region in the country, Northeastern India has not undergone any investigation regarding the presence of T. gondii and Trichinella spp. in pigs. Therefore, this study aims to determine the seroprevalence of T. gondii and Trichinella spp. and identify associated risk factors in pigs reared by tribal communities and small-holder livestock farmers in the northeastern region of India. In a cross-sectional serological survey, 400 pigs from 400 households across five northeastern states of India underwent testing for the seroprevalence of porcine toxoplasmosis and trichinellosis. Serum samples (80 from each state) were analyzed using commercially available ELISA assays. Data on backyard farm characteristics and various management aspects were collected, and risk factors linked with prevalence were analyzed through univariate and multivariate logistic regression analysis. The findings revealed that the apparent and true prevalence of anti-T. gondii antibodies were 45% (40.12-49.88, 95% CI) and 45.7% (40.7-50.69, 95% CI), respectively. As for anti- Trichinella antibodies, both the apparent and true prevalence were 0.75% (-0.1-1.6, 95% CI). The univariate and multivariate analyses indicated that age above 24 months (OR 7.20, 95% CI 2.45-23.71), exposure to cats (OR = 5.87, 95% CI 2.55-14.05), and farms operating for breeding purposes (OR = 5.60, 95% CI 3.01-11.04) were significant risk factors associated with the seroprevalence of T. gondii. This study marks the initial documentation of the seroprevalence of T. gondii and Trichinella spp. in pigs reared by tribal communities in Northeastern India. The results emphasize the significance of these parasites as foodborne zoonotic threats in the region, potentially posing substantial public health risks, especially within tribal and rural communities. The insights derived from this research could be valuable in formulating targeted preventive and control strategies against T. gondii and Trichinella spp. in pigs, not only in this region but also in areas with similar rearing practices.

First detection of Toxoplasma gondii Africa 4 lineage in a population of carnivores from South Africa.

Introduction: There have only been a few molecular studies conducted on the detection of T. gondii in tissues of carnivores in South Africa, with no data on the genetic diversity of this parasite. That is why the aim of this study was to detect and genotype T. gondii DNA in tissues of selected wild and domestic carnivores in South Africa. Methods: Samples were collected from 80 animals of 20 species (mainly road-killed) in the four provinces of Limpopo (n=57), Mpumalanga (n=21), Gauteng (n=1) and Free State (n=1) during the period 2014-2018. Samples of brain (n=31), heart (n=4), liver (n=40), spleen (n=2) and lung (n=3) were used to detect T. gondii by real-time PCR targeting a 529 bp repeating fragment of T. gondii DNA. Samples that were positive in real-time PCR were genotyped using 15 microsatellite markers. Results: T. gondii DNA was detected in 4 (5 %) samples: in the brain from a Black-backed Jackal (Canis mesomelas), in the liver from a African Wildcat (Felis silvestris lybica) and in the liver and heart of two Rusty-spotted Genets (Genetta maculata) respectively. The DNA sample from Black-backed Jackal was genotyped and characterized as belonging to the type Africa 4 lineage (equivalent to RFLP genotype ToxoDB#20), that is a widespread lineage in Africa. Discussion: This is the first genetic characterization of T. gondii isolated from a wild carnivore on the African continent and the first report of T. gondii in Black-backed Jackal. The Africa 4 lineage was also confirmed in the region of Southern Africa for the first time.

The role of species ecology in predicting Toxoplasma gondii prevalence in wild and domesticated mammals globally.

Macroecological approaches can provide valuable insight into the epidemiology of globally distributed, multi-host pathogens. Toxoplasma gondii is a protozoan that infects any warm-blooded animal, including humans, in almost every habitat worldwide. Toxoplasma gondii infects its hosts through oocysts in the environment, carnivory of tissue cysts within intermediate host prey and vertical transmission. These routes of infection enable specific predictions regarding the ecological and life history traits that should predispose specific taxa to higher exposure and, thus infection rates of T. gondii. Using T. gondii prevalence data compiled from 485 studies representing 533 free-ranging wild mammalian species, we examined how ecological (habitat type, trophic level) and life history (longevity, vagility, gestation duration and torpor) traits influence T. gondii infection globally. We also compared T. gondii prevalence between wild and domesticated species from the same taxonomic families using data compiled from 540 studies of domestic cattle, sheep, and pigs. Across free-ranging wildlife, we found the average T. gondii prevalence was 22%, which is comparable to the global human estimate. Among ecological guilds, terrestrial species had lower T. gondii prevalence than aquatic species, with freshwater aquatic taxa having an increased prevalence compared to marine aquatic species. Dietary niches were also influential, with carnivores having an increased risk compared to other trophic feeding groups that have reduced tissue cyst exposure in their diet. With respect to influential life history traits, we found that more vagile wildlife species had higher T. gondii infection rates, perhaps because of the higher cumulative risk of infection during movement through areas with varying T. gondii environmental loads. Domestic farmed species had a higher T. gondii prevalence compared to free-ranging confamilial wildlife species. Through a macroecological approach, we determined the relative significance of transmission routes of a generalist pathogen, demonstrating an increased infection risk for aquatic and carnivorous species and highlighting the importance of preventing pathogen pollution into aquatic environments. Toxoplasma gondii is increasingly understood to be primarily an anthropogenically-associated pathogen whose dissemination is enhanced by ecosystem degradation and human subsidisation of free-roaming domestic cats. Adopting an ecosystem restoration approach to reduce one of the world's most common parasites would synergistically contribute to other initiatives in conservation, feline and wildlife welfare, climate change, food security and public health.

Toxoplasma gondii Infection in the Eurasian Beaver (Castor fiber) in Switzerland: Seroprevalence, Genetic Characterization, and Clinicopathologic Relevance.

Toxoplasma gondii is a coccidian parasite able to infect all warm-blooded animals and humans. Rodents are one of the most important intermediate hosts for T. gondii, but little is known about infection in beavers and its clinical relevance. Toxoplasmosis was not considered an important waterborne disease until recently, but with increased outbreaks in humans and animals this perspective has changed. Serum samples from 247 Eurasian beavers (Castor fiber) collected from 2002 to 2022 were tested for antibodies to T. gondii by a commercial ELISA. Antibodies to T. gondii were found in 113 (45.8%) beavers. Higher weight and proximity to urban areas were found to be significant predictors for seropositivity. Additionally, T. gondii DNA was detected in 23/41 brain tissue samples by real-time PCR. Histopathologic examination of brain sections revealed inflammatory changes in 26/40 beavers, mainly characterized by encephalitis, meningitis, choroid plexitis, or a combination of them. In six of these cases the lesions were in direct association with parasitic stages. With an adapted nested PCR multilocus sequence typing and in silico restriction fragment length polymorphism analysis approach, three different T. gondii genotypes were detected in brain samples: the clonal Type II strain (ToxoDB 1), a Type II variant (ToxoDB 3), and a novel genotype exhibiting both Type II and I alleles in a further animal. Toxoplasma gondii infections in beavers have epidemiologic and clinical significance. The high seroprevalence indicates frequent contact with the parasite, and as competent intermediate hosts they may play an important role, contributing to maintaining the life cycle of T. gondii in semiaquatic habitats. In addition, although most beavers appear to develop subclinical to chronic disease courses, acute and fatal outcomes, mainly characterized by encephalitis and generalized infection, do also occur.

Distribution and prevalence of antibodies to Trichinella spp. and Toxoplasma gondii in wild pigs (Sus scrofa) in the United States.

Invasive wild pigs (Sus scrofa) are a reservoir for over 100 viral, bacterial, and parasitic pathogens that are transmissible to humans, livestock, domestic animals, and wildlife in North America. Numerous historical local surveys and results from a nation-wide survey (2006-2010) indicated that wild pigs in the United States act as reservoirs for Trichinella spp. and Toxoplasma gondii, two zoonotic pathogens of importance for human and animal health. Since that time, wild pig populations have expanded and increased in density in many areas. Population expansion of wild pigs creates opportunities for the introduction of pathogens to new areas of the country, increasing health risks. The goal of this study was to investigate the current geographic distribution and prevalence of Trichinella spp. and T. gondii antibodies in wild pigs using serum samples collected from 2014 to 2020. Serum samples from 36 states were tested for antibodies to Trichinella spp. (n = 7467) and T. gondii (n = 5984) using commercially available enzyme-linked immunosorbent assays. Seroprevalence for Trichinella spp. (12.4%, 927/7467) and T. gondii (40.8%, 2444/5984) are significantly higher compared to a previous 2006-2010 study across all regions. Results from this study also showed a lower seroprevalence (4.8%) for Trichinella spp. in the West region compared to the other regions (South: 13.4%; Midwest: 18.4%; Northeast: 19.1%). There were new detection records for antibodies to Trichinella spp. in 11 states, mostly in the West, Midwest, and Northeast regions compared to a previous study in 2014. Males and juveniles were less likely to be positive for Trichinella spp. antibodies, compared to females and older animals, respectively. Seroprevalence was similar for T. gondii across the regions (31.8-56%) with some states having particularly high seroprevalence (e.g., Hawaii 79.4% and Pennsylvania 68%). There were new T. gondii antibody detection records for 12 states, mostly in the West, Midwest, and Northeast regions. Adults were more likely than juveniles and subadults to be seropositive. These data confirm that the distribution and prevalence of antibodies for Trichinella spp. and T. gondii are increasing in the United States, likely driven by wild pig population growth and range expansion.

Seroprevalence and risk factors associated with Toxoplasma gondii in pigs in Haryana, India.

AIMS: Toxoplasmosis is one of the most common food-borne parasitic zoonosis, caused by Toxoplasma gondii, an obligate intracellular protozoan parasite. A cross-sectional study was carried out to determine the seroprevalence of T. gondii and associated risk factors in pigs in Haryana, India. METHODS AND RESULTS: Serum samples were collected from 429 pigs from three agroclimatic zones (I-III) of Haryana and analysed for the presence of antibodies against T. gondii using a commercial indirect enzyme-linked immunosorbent assay (ELISA). Anti-T. gondii antibodies were detected in 106 animals (24.7%), with the highest seropositivity in zone II (31.3%) followed by zone III (24.4%) and zone I (18.3%). Risk factors associated with higher seropositivity in pigs were farm size (higher in large-sized farms), age (higher in pigs >1 year of age), sex (higher in males), type of feeding (higher in combination of homemade and hotel waste) and housing (higher in free-ranging pigs). CONCLUSIONS: The findings of the study testify to the exposure of pigs (of all agro-climatic zones) to T. gondii. Hence, the observations are of significant medical and veterinary importance for devising and implementing control measures to check the dissemination of toxoplasmosis to pigs and eventually to humans.

Isolation of Toxoplasma gondii in cell culture: an alternative to bioassay.

Toxoplasma gondii is an apicomplexan protozoan parasite that can infect mammals and birds. The infection can cause acute toxoplasmosis and death in susceptible hosts. Bioassay using cats and mice has been the standard for the isolation of T. gondii from infected hosts for the past several decades. However, bioassay is labor-intensive, expensive, and involves using laboratory animals. To search alternative approaches and o work towards replacement of animal experiments, we summarized the key literature and conducted four experiments to isolate T. gondii in vitro by cell culture. A few heart tissue samples from animals with the highest antibody titers in a given collection were used for T. gondii isolation. These experiments included samples from five out of 51 wild ducks, four of 46 wild turkeys, six of 24 white-tailed deer, as well as from six kangaroos that had died with acute toxoplasmosis in a zoo. These experiments resulted in three isolates from five chronically infected wild ducks (60%), four isolates from four chronically infected wild turkeys (100%), one isolate from six chronically infected white-tailed deer (17%), and four isolates from six kangaroos with acute toxoplasmosis (67%). In addition, five isolates from the five chronically infected wild ducks were obtained by bioassay in mice, showing a 100% success rate, which is higher than the 60% rate by direct cell culture. These T. gondii isolates were successfully propagated in human foreskin fibroblast (HFF) or Vero cells, and genotyped by multilocus PCR-RFLP markers. The results showed that it is practical to isolate T. gondii directly in cell culture. Although the cell culture approach may not be as sensitive as the bioassay, it does provide an alternative that is simple, cost-effective, ethically more acceptable, and less time-sensitive to isolate T. gondii. In this paper we propose a procedure that may be applied and further optimized for isolation of T. gondii.

Molecular characterization and epidemiology of Toxoplasma gondii isolates from free-range chickens in the southwest region of Goiás: new genotypes.

The purpose of this study was to isolate Toxoplasma gondii from tissues of free-range chickens in the southwestern region of Goiás, to detect and molecularly characterize the genetic material of the parasite, and to determine the seroprevalence of the protozoan parasite in these animals. A seroprevalence of T. gondii antibodies of 76% (19/25) was found among the chickens, while genetic material from their tissues was detected in 56% (14/25). A total of 14 isolates was obtained in the bioassay, ten of which were considered acute, eight were considered isolates of high virulence lethal to mice, and four of low virulence, considered non-lethal but with the ability to chronify the infection. Seven of the ten isolates showed significant morphometric differences from the RH strain, in terms of nucleus-complex-apical distance, length and width. Genotyping of the acute isolates was performed by RFLP-PCR, using 11 genetic markers: SAG1, SAG2 (3'SAG2 and 5'SAG2), alt.SAG2, SAG3, BTUB, GRA6, c22-8, c29-2, L358, PK1, and APICO. The results were compared and classified according to the genotypes listed on the ToxoDB Platform, where different profiles were observed indicating the presence of two known genotypes (#7 and #63) and five new genotypes (NEW 3, NEW4, NEW5, NEW6, NEW 7). The results showed high seroprevalence, isolation rate, molecular detection and genotypic variations of T. gondii in free-range chickens in the southwestern region of Goiás.

Global epidemiology and spatial distribution of Toxoplasma gondii in goats: Protocol for a systematic review and Bayesian hierarchical meta-analysis.

BACKGROUND: Toxoplasma gondii, a cosmopolitan protozoan parasite causes toxoplasmosis in humans and many species of domestic and wild animals. T. gondii instigates significant economic losses in sheep and goat farming industry and can lead to abortion, stillbirth, congenital malformations and neonatal losses. The objective of this protocol is to evaluate worldwide seroprevalence of T. gondii exposure in goats using Bayesian hierarchical meta-analysis and geographic information system (GIS). METHODS: A comprehensive literature search will be conducted using search engines, including Web of Science, ScienceDirect, Scopus, PubMed, ProQuest, EMBASE, PROSPERO Register and, Google Scholar without date and language restrictions. The authors search for cross-sectional studies that determine the seroprevalence of T. gondii in goats. Two reviewers will independently screen, selected studies; also, they will extract data, and assess the risk of bias. In case(s) of disagreement, a consensus will be reached with the help of a third author. The Bayesian hierarchical meta-analysis will use to estimate country and worldwide true seroprevalence of T. gondii, which is consist of the sensitivity and specificity of the applied serological assays. The obtained data will be used to identify country-level risk factors associated with T. gondii exposure using GIS in the ArcGIS software. DISCUSSION: The systematic review produced from this protocol will provide the true prevalence rate and spatial distribution T. gondii exposure in goats both regionally and globally using Bayesian hierarchical and GIS analysis. SYSTEMATIC REVIEW REGISTRATION: PROSPERO registration number: CRD42020107928.

Molecular confirmation of the systemic toxoplasmosis in cat.

INTRODUCTION AND OBJECTIVE: Systemic toxoplasmosis with tissue-spread parasites occurring in intermediate hosts may also occur in immunocompromised cats (e.g., infected with FLV or FIV). To the best of our knowledge, no reports have been published on the detection and genotyping of T. gondii DNA in cats with extraintestinal toxoplasmosis in Poland. The article describes the case of the sudden death of 3 out of 4 cats in a cattery, and the detection and molecular characterization of T. gondii DNA detected in the tissues of one of the dead cats. MATERIAL AND METHODS: Samples of brain, lungs, heart, and liver of the cat that died suddenly were examined for the presence of T. gondii DNA (B1 gene) by nested PCR and real-time PCR. DNA positive samples were also genotyped at 12 genetic markers using multiplex multilocus nested PCR-RFLP (Mn-PCR-RFLP) and multilocus sequence typing (MLST). RESULTS: A total of 9 out of the 20 DNA samples were successfully amplified with nested and/or Real-time PCR. DNA from 3 out of 5 types of tested samples were genotyped (brain, heart and muscle). Mn-PCR-RFLP and MLST results revealed type II (and II/III at SAG1) alleles at almost all loci, except a clonal type I allele at the APICO locus. This profile corresponds to the ToxoDB#3 genotype, commonly identified amongst cats in Central Europe. CONCLUSIONS: To the best of our knowledge, this is the first study describing the genetic characteristics of T. gondii population determined in a cat in Poland. These data confirm the importance of this host as a reservoir for this pathogen, and demonstrate the genotypic variation of this parasite. Veterinarians should take into account that cats may develop disseminated toxoplasmosis, and that it is a systemic disease which may lead to the death of the cat, and to transmission of the pathogen to other domestic animals and to humans.

Genetic diversity of Toxoplasma gondii in South America: occurrence, immunity, and fate of infection.

Toxoplasma gondii is an intracellular parasite with a worldwide distribution. Toxoplasma gondii infections are of great concern for public health, and their impact is usually most severe in pregnant women and their foetuses, and in immunocompromised individuals. Displaying considerable genetic diversity, T. gondii strains differ widely according to geographical location, with archetypal strains predominantly found in the Northern Hemisphere and non-archetypal (atypical) strains, with highly diverse genotypes, found mainly in South America. In this review, we present an overview of the identification and distribution of non-archetypal strains of T. gondii. Special attention is paid to the strains that have been isolated in Brazil, their interaction with the host immunological response, and their impact on disease outcomes. The genetic differences among the strains are pivotal to the distinct immunological responses that they elicit. These differences arise from polymorphisms of key proteins released by the parasite, which represent important virulence factors. Infection with divergent non-archetypal strains can lead to unusual manifestations of the disease, even in immunocompetent individuals.

High prevalence and diversity of Toxoplasma gondii DNA in feral cat feces from coastal California.

Toxoplasma gondii is a zoonotic parasite that can cause severe morbidity and mortality in warm-blooded animals, including marine mammals such as sea otters. Free-ranging cats can shed environmentally resistant T. gondii oocysts in their feces, which are transported through rain-driven runoff from land to sea. Despite their large population sizes and ability to contribute to environmental oocyst contamination, there are limited studies on T. gondii oocyst shedding by free-ranging cats. We aimed to determine the frequency and genotypes of T. gondii oocysts shed by free-ranging domestic cats in central coastal California and evaluate whether genotypes present in feces are similar to those identified in sea otters that died from fatal toxoplasmosis. We utilized a longitudinal field study of four free-ranging cat colonies to assess oocyst shedding prevalence using microscopy and molecular testing with polymerase chain reaction (PCR). T. gondii DNA was confirmed with primers targeting the ITS1 locus and positive samples were genotyped at the B1 locus. While oocysts were not visualized using microscopy (0/404), we detected T. gondii DNA in 25.9% (94/362) of fecal samples. We genotyped 27 samples at the B1 locus and characterized 13 of these samples at one to three additional loci using multi locus sequence typing (MLST). Parasite DNA detection was significantly higher during the wet season (16.3%, 59/362) compared to the dry season (9.7%; 35/362), suggesting seasonal variation in T. gondii DNA presence in feces. High diversity of T. gondii strains was characterized at the B1 locus, including non-archetypal strains previously associated with sea otter mortalities. Free-ranging cats may thus play an important role in the transmission of virulent T. gondii genotypes that cause morbidity and mortality in marine wildlife. Management of free-ranging cat colonies could reduce environmental contamination with oocysts and subsequent T. gondii infection in endangered marine mammals and people.

Seroprevalence and associated factors with Neospora caninum infection in sheep and goats slaughtered in the state of Paraíba, Brazil.

The objective was to describe the seroprevalence of anti-Neospora caninum antibodies in goats and sheep slaughtered in the state of Paraíba and to identify possible associated factors with the infections. Two hundred twenty-nine samples from goats and two hundred five from sheep were analyzed by Indirect Immunofluorescence Reaction (IFAT) using a cutoff point of 1:50. The presence of anti-N. caninum antibodies was identified in 28.4% (65/ 229; 95% Confidence Interval: 22.6-34.2) of the goat samples and in 12.7% (26/ 205; 95% CI: 8.2 - 17.2) of the sheep samples. Contact between goats and dogs (Odds ratio 4.81; CI 1.13 - 2.67; p = 0.041) and cattle (OR. 1.87; CI 1.13 - 2.67; p = 0.002) was identified as a risk factor for goats and contact between sheep and dogs (OR 2.32; CI 1.58 - 3.14; p = 0.026) and history of abortion (OR 1.94; CI 1.28 - 2.90; p = 0.001) was considered a risk factor for sheep. The study revealed a high seroprevalence of anti-N. caninum antibodies in slaughtered goats and sheep in Paraíba. Risk factors such as contact with dogs/cattle and abortion history underscore the need for preventive measures to control infection and enhance animal health management.