Direct evidence of cheetah (Acinonyx jubatus) as intermediate host of Toxoplasma gondii through isolation of viable strains.

Toxoplasma gondii causes lifelong infection in most definitive and intermediate hosts. Clinical cases of toxoplasmosis in captive cheetahs have been reported. However, there are few reports of viable T. gondii strains isolated from cheetahs. Here, T. gondii infection was investigated using molecular and serological assays in cheetahs from China. Modified agglutination test (MAT) (cut-off: 1:25) indicated that all six examined cheetahs (n = 6) showed T. gondii antibodies. Toxoplasma gondii DNA was detected in three out of five cheetahs. Two viable T. gondii strains were isolated from the striated muscles of two cheetahs using mice bioassay. They were designated as TgCheetahCHn1 and TgCheetahCHn2. Genetic characterization of DNA derived from tachyzoites was performed using RFLP-PCR of 10 markers. Toxoplasma gondii TgCheetahCHn1 is ToxoDB PCR-RFLP genotype #319, and the alleles of ROP18/ROP5 types were 3/7. TgCheetahCHn2 is ToxoDB genotype #9, and the alleles of ROP18/ROP5 were 3/6. The average survival time of TgCheetahCHn1-infected Swiss mice was 22 ± 1 days (n = 23), and the mice did not have detectable T. gondii-specific antibodies until 117 ± 30 days post-inoculation (n = 8), therefore, TgCheetahCHn1 had intermediate virulence. TgCheetahCHn2 was avirulent for Swiss mice. Few brain tissue cysts (0-50) were observed in the mice inoculated with TgCheetahCHn1 or TgCheetahCHn2. The results provide direct evidence of cheetah as intermediate host of T. gondii.

Toxoplasma gondii Survey in Waterfowl and Gulls from Eight USA States.

Sera from 391 waterbirds from eight USA states were tested for Toxoplasma gondii antibodies using the modified agglutination test. Fifteen different waterbird species (26.6%; n=104) were seropositive. Of the adults, 25.4% (n=52) showed a significantly higher T. gondii seroprevalence compared with juveniles (13.4%; n=17); however, sex was not a significant factor.

From TgO/GABA-AT, GABA, and T-263 Mutant to Conception of Toxoplasma.

Toxoplasma gondii causes morbidity, mortality, and disseminates widely via cat sexual stages. Here, we find T. gondii ornithine aminotransferase (OAT) is conserved across phyla. We solve TgO/GABA-AT structures with bound inactivators at 1.55 Å and identify an inactivator selective for TgO/GABA-AT over human OAT and GABA-AT. However, abrogating TgO/GABA-AT genetically does not diminish replication, virulence, cyst-formation, or eliminate cat's oocyst shedding. Increased sporozoite/merozoite TgO/GABA-AT expression led to our study of a mutagenized clone with oocyst formation blocked, arresting after forming male and female gametes, with "Rosetta stone"-like mutations in genes expressed in merozoites. Mutations are similar to those in organisms from plants to mammals, causing defects in conception and zygote formation, affecting merozoite capacitation, pH/ionicity/sodium-GABA concentrations, drawing attention to cyclic AMP/PKA, and genes enhancing energy or substrate formation in TgO/GABA-AT-related-pathways. These candidates potentially influence merozoite's capacity to make gametes that fuse to become zygotes, thereby contaminating environments and causing disease.

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.

Epidemiology and isolation of viable Toxoplasma gondii strain from macropods.

Wallabies and other marsupials are highly susceptible to Toxoplasma gondii. In this study, 26 macropod samples were collected (8 red kangaroos, 4 Parma wallabies, 8 red-necked wallabies, 5 albino red-necked wallabies and 1 Eastern grey kangaroo), including tissue (n = 9) and serum (n = 17) samples. According to the modified agglutination test (MAT) results (cutoff 1:25), 50% (95% Cl: 32.06-67.94%) of the macropods had T. gondii antibodies. Among them, species, survival state, and sampling date were risk factors for T. gondii susceptibility (P < 0.05). T. gondii DNA was detected in two (cases #14 and #15) of the nine cases obtained from macropod tissues. One viable T. gondii strain (TgRooCHn4) was isolated from an albino red-necked wallaby (Macropus rufogriseus, case #14) via bioassay in mice. TgRooCHn4 belongs to ToxoDB genotype #3, using the 10 multilocus PCR-RFLP markers. The ROP18 and ROP5 gene allele types of TgRooCHn4 were 2/2, which was predicted to be non-lethal to mice. The virulence of TgRooCHn4 tachyzoites was avirulent in mice. Most macropods sampled from Hernan province in 2021 and 2022 were positive with T. gondii infection. A flood occurred in July 2021 in Zhengzhou from Henan province may promote the transmission of T. gondii oocysts. To our knowledge, this is the first T. gondii strain isolated from albino red-necked wallaby. However, further investigation is required to enhance our understanding of the transmission and prevention of toxoplasmosis in sensitive zoo animals.

Two viable Toxoplasma gondii isolates from red-necked wallaby (Macropus rufogriseus) and red kangaroo (M. rufus).

Wallabies and kangaroos are susceptible to Toxoplasma gondii. However, little information concerning T. gondii infection in captive macropods is available. Three dead macropods collected from a zoo exhibited no clinical symptoms associated with toxoplasmosis. Heart fluids were tested for T. gondii antibodies using a modified agglutination test. T. gondii DNA samples derived from macropod tissues were tested by Polymerase Chain Reaction. Viable T. gondii were isolated from myocardium of macropods via mouse bioassay. Tissues (brain, lungs, or mesenteric lymph nodes) from T. gondii-positive mice were seeded into Vero cell culture flasks. The virulence of the isolated T. gondii strains was evaluated in Swiss mice. The DNA from T. gondii tachyzoites obtained from cell cultures was characterized by 10 PCR-RFLP markers and the virulence genes, ROP18 and ROP5. T. gondii antibodies were identified in two of the three macropods (Macropod#5 and #7). T. gondii DNA was obtained from the heart and lungs of Macropod#7. Two viable T. gondii strains were isolated from the myocardium of Macropus rufogriseus (Macropod#5) and M. rufus (Macropod#7) via mouse bioassay and designated as TgRooCHn2 and TgRooCHn3, respectively. TgRooCHn2 was ToxoDB genotype#3, and TgRooCHn3 was ToxoDB genotyp#2. Both 104 TgRooCHn2 and TgRooCHn3 tachyzoites had intermediate virulence in mice. M. rufogriseus (Macropod#5) and M. rufus (Macropod#7) may have been in the initial stages of toxoplasmosis, due to a recent T. gondii infection with oocysts. This study is the first to document the T. gondii ToxoDB#3 isolate in macropods. T. gondii infection in captive macropods indicates the urgent need to control the transmission of this parasite in the environment, food and water of zoo animals.

Additional evidence of tigers (Panthera tigris altaica) as intermediate hosts for Toxoplasma gondii through the isolation of viable strains.

Toxoplasmosis is one of the most common zoonotic diseases in the world. Felines excrete Toxoplasma gondii oocysts, which play a key role in the transmission of this protozoon. Pathological diagnoses were performed on four carcasses of captive tigers collected from 2019 to 2021 in China, and T. gondii was surveyed using serology, molecular analysis, and aetiology. Striated muscle samples of the tigers (n = 4) were bioassayed in mice. DNA derived from T. gondii tachyzoites was isolated and characterized using PCR-RFLP. The pathological diagnoses revealed that ageing, declined immune function, liver, and kidney failures caused the deaths in the tigers examined. A modified agglutination test (cut-off: 1:25) revealed that IgG antibodies to T. gondii were 100% (4/4) in the captive tigers. Two viable T. gondii strains (TgTigerCHn3 and TgTigerCHn4) were isolated from tiger striated muscles and seeded on the Vero cell culture for further propagation. The genotypes of TgTigerCHn3 and TgTigerCHn4 were ToxoDB#20 and ToxoDB#2, respectively. The two strains were avirulent for Swiss mice, which matched the ROP18 and ROP5 gene alleles of TgtigerCHn3 (3/4) and TgtigerCHn4 (3/3). Few brain tissue cysts (0-213) were observed in the mice after inoculation with TgTigerCHn3 and TgTigerCHn4. This is the first documented isolation of T. gondii ToxoDB#20 and ToxoDB#2 from tigers. The results provide additional direct evidence of tiger as intermediate hosts for T. gondii. Tigers in the zoos may potentially transmit T. gondii to other animals and humans.

Prevalence and genotype of Toxoplasma gondii in stranded Hawaiian cetaceans.

Toxoplasma gondii is a significant threat to endangered Hawaiian wildlife including birds and marine mammals. To estimate the prevalence of T. gondii in stranded cetaceans from 1997 to 2021 in Hawai'i, we tested tissues from 37 stranded spinner dolphins Stenella longirostris and 51 stranded individuals that represented 18 other cetacean species. DNA from cetacean tissue extracts were screened using a nested polymerase chain reaction (PCR) assay targeting the Toxoplasmatinae internal transcribed spacer 1 of the nuclear ribosomal DNA. A positive result was obtained in 9 tissues examined for each of 2 spinner dolphins out of 525 tissue samples analyzed by PCR. The PCR-positive spinner dolphins had disseminated acute toxoplasmosis with necrosis, inflammation, and intralesional protozoal cysts and tachyzoites in multiple organs. Discrete positive immunostaining for T. gondii was observed in all tissues tested including the adrenal gland, brain, liver, and lung. Both positive spinner dolphins were negative for cetacean morbillivirus. The T. gondii genotyping was performed by restriction fragment length polymorphism (PCR-RFLP) based on 10 genetic markers. The PCR-RFLP analysis revealed the T. gondii belonged to PCR-RFLP-ToxoDB genotype #24, previously detected in wild pig Sus scrofa in O'ahu, bobcats Lynx rufus from Mississippi, USA, and chickens Gallus gallus from Costa Rica and Brazil. These cases represent the first report of this genotype in aquatic mammals and the second and third reports of fatal disseminated T. gondii infection in stranded spinner dolphins from Hawai'i. Nearshore species, like spinner dolphins, may be at increased risk of mortality from this parasite in marine coastal waterways via sewage systems, storm water drainage, and freshwater runoff.

Toxoplasma gondii clonal type III is the dominant genotype identified in Grenadian pigs.

BACKGROUND: Toxoplasma gondii is a widespread zoonotic protozoan parasite capable of infecting all warm-blooded animals. Although the genotypes of T. gondii in pigs have been reported worldwide, there is no information on the genotypes and diversity of T. gondii in pigs in Grenada, West Indies. OBJECTIVES: The aims of the present study were to isolate, genotype and determine the diversity of T. gondii genotypes in pigs. METHODS: We carried out a modified agglutination test (MAT) on blood from 149 pig hearts collected from a local meat market. Myocardial tissue homogenate from pigs that tested positive for T. gondii was homogenized and inoculated into mice for isolation of the parasite. We collected mouse tissues and extracted DNA for genotyping based on 11 polymerase chain reaction-restriction fragment length polymorphism markers (SAG1, SAG2, alt. SAG2, SAG 3, BTUB, GRA6, L358, PK1, C22-8, C 29-2 and Apico). RESULTS: Out of the 149 pig hearts, 31 (20.8%) tested positive for T. gondii on MAT. Bioassays in mice yielded 12 isolates designated TgpgGr1 to TgpgGr12. Molecular characterisation of T. gondii revealed four genotypes as follows: ToxoDB #2-clonal type III (seven isolates); ToxoDB #7 (three isolates); ToxoDB #13 (one isolate); ToxoDB #30 (1 isolate). Overall, ToxoDB #2 was the most common (58%). Toxo database (DB) # 13, which causes interstitial pneumonia in affected mice, has also been reported. CONCLUSION: The genetic diversity of T. gondii in pigs in Grenada is lower than that in other surrounding Caribbean areas.

Unifying Virulence Evaluation in Toxoplasma gondii: A Timely Task.

Toxoplasma gondii, a major zoonotic pathogen, possess a significant genetic and phenotypic diversity that have been proposed to be responsible for the variation in clinical outcomes, mainly related to reproductive failure and ocular and neurological signs. Different T. gondii haplogroups showed strong phenotypic differences in laboratory mouse infections, which provide a suitable model for mimicking acute and chronic infections. In addition, it has been observed that degrees of virulence might be related to the physiological status of the host and its genetic background. Currently, mortality rate (lethality) in outbred laboratory mice is the most significant phenotypic marker, which has been well defined for the three archetypal clonal types (I, II and III) of T. gondii; nevertheless, such a trait seems to be insufficient to discriminate between different degrees of virulence of field isolates. Many other non-lethal parameters, observed both in in vivo and in vitro experimental models, have been suggested as highly informative, yielding promising discriminatory power. Although intra-genotype variations have been observed in phenotypic characteristics, there is no clear picture of the phenotypes circulating worldwide; therefore, a global overview of T. gondii strain mortality in mice is presented here. Molecular characterization has been normalized to some extent, but this is not the case for the phenotypic characterization and definition of virulence. The present paper proposes a baseline (minimum required information) for the phenotypic characterization of T. gondii virulence and intends to highlight the needs for consistent methods when a panel of T. gondii isolates is evaluated for virulence.

Are foxes (Vulpes spp.) good sentinel species for Toxoplasma gondii in northern Canada?

BACKGROUND: In changing northern ecosystems, understanding the mechanisms of transmission of zoonotic pathogens, including the coccidian parasite Toxoplasma gondii, is essential to protect the health of vulnerable animals and humans. As high-level predators and scavengers, foxes represent a potentially sensitive indicator of the circulation of T. gondii in environments where humans co-exist. The objectives of our research were to compare serological and molecular assays to detect T. gondii, generate baseline data on T. gondii antibody and tissue prevalence in foxes in northern Canada, and compare regional seroprevalence in foxes with that in people from recently published surveys across northern Canada. METHODS: Fox carcasses (Vulpes vulpes/Vulpes lagopus, n = 749) were collected by local trappers from the eastern (Labrador and Québec) and western Canadian Arctic (northern Manitoba, Nunavut, and the Northwest Territories) during the winters of 2015-2019. Antibodies in heart fluid were detected using a commercial enzyme-linked immunosorbent assay. Toxoplasma gondii DNA was detected in hearts and brains using a magnetic capture DNA extraction and real-time PCR assay. RESULTS: Antibodies against T. gondii and DNA were detected in 36% and 27% of foxes, respectively. Detection of antibodies was higher in older (64%) compared to younger foxes (22%). More males (36%) than females (31%) were positive for antibodies to T. gondii. Tissue prevalence in foxes from western Nunavik (51%) was higher than in eastern Nunavik (19%). At the Canadian scale, T. gondii exposure was lower in western Inuit regions (13%) compared to eastern Inuit regions (39%), possibly because of regional differences in fox diet and/or environment. Exposure to T. gondii decreased at higher latitude and in foxes having moderate to little fat. Higher mean infection intensity was observed in Arctic foxes compared to red foxes. Fox and human seroprevalence showed similar trends across Inuit regions of Canada, but were less correlated in the eastern sub-Arctic, which may reflect regional differences in human dietary preferences. CONCLUSIONS: Our study sheds new light on the current status of T. gondii in foxes in northern Canada and shows that foxes serve as a good sentinel species for environmental circulation and, in some regions, human exposure to this parasite in the Arctic.

Possible Cross-Reactivity of Feline and White-Tailed Deer Antibodies against the SARS-CoV-2 Receptor Binding Domain.

In late 2019, a novel coronavirus began circulating within humans in central China. It was designated SARS-CoV-2 because of its genetic similarities to the 2003 SARS coronavirus (SARS-CoV). Now that SARS-CoV-2 has spread worldwide, there is a risk of it establishing new animal reservoirs and recombination with native circulating coronaviruses. To screen local animal populations in the United States for exposure to SARS-like coronaviruses, we developed a serological assay using the receptor binding domain (RBD) from SARS-CoV-2. SARS-CoV-2's RBD is antigenically distinct from common human and animal coronaviruses, allowing us to identify animals previously infected with SARS-CoV or SARS-CoV-2. Using an indirect enzyme-linked immunosorbent assay (ELISA) for SARS-CoV-2's RBD, we screened serum from wild and domestic animals for the presence of antibodies against SARS-CoV-2's RBD. Surprisingly prepandemic feline serum samples submitted to the University of Tennessee Veterinary Hospital were ∼50% positive for anti-SARS RBD antibodies. Some of these samples were serologically negative for feline coronavirus (FCoV), raising the question of the etiological agent generating anti-SARS-CoV-2 RBD cross-reactivity. We also identified several white-tailed deer from South Carolina with anti-SARS-CoV-2 antibodies. These results are intriguing, as cross-reactive antibodies toward SARS-CoV-2 RBD have not been reported to date. The etiological agent responsible for seropositivity was not readily apparent, but finding seropositive cats prior to the current SARS-CoV-2 pandemic highlights our lack of information about circulating coronaviruses in other species. IMPORTANCE We report cross-reactive antibodies from prepandemic cats and postpandemic South Carolina white-tailed deer that are specific for that SARS-CoV RBD. There are several potential explanations for this cross-reactivity, each with important implications to coronavirus disease surveillance. Perhaps the most intriguing possibility is the existence and transmission of an etiological agent (such as another coronavirus) with similarity to SARS-CoV-2's RBD region. However, we lack conclusive evidence of prepandemic transmission of a SARS-like virus. Our findings provide impetus for the adoption of a One Health Initiative focusing on infectious disease surveillance of multiple animal species to predict the next zoonotic transmission to humans and future pandemics.

Isolation, genotyping and virulence determination of a Toxoplasma gondii strain from non-human primate from China.

Toxoplasma gondii infects almost all warm-blooded animals, including humans and non-human primates. Many cases of T. gondii infection in non-human primates have been reported worldwide. In this study, 15 monkeys were collected from zoos in Henan Province between 2016 and 2019. A modified agglutination test (MAT) (cut-off: 1:8) showed that 46.7% (7/15) of the heart juices had T. gondii IgG antibody transformation. One viable T. gondii strain was successfully isolated from the myocardium of a rhesus monkey by bioassay in mice. This strain was designated as TgMonkeyCHn1. The DNA of T. gondii tachyzoites was obtained using cell cultures, and the genotype of this strain was determined by PCR-RFLP with 10 markers and the virulence genes ROP5 and ROP18. The genotype and ROP18/ROP5 (3/6) of TgMonkeyCHn1 did not match any known genotypes. In addition, the TgMonkeyCHn1 formed low number of tissue cysts and was non-lethal to mice. To our knowledge, this is the first T. gondii strain isolated from Old World monkeys. Rhesus monkey is a new host record for T. gondii.

Toxoplasma gondii contamination at an animal agriculture facility: Environmental, agricultural animal, and wildlife contamination indicator evaluation.

Toxoplasma gondii is a parasite of significant public health importance. We attempted to detect T. gondii contamination and assess advantages and disadvantages of contamination indicators through surveilling soil, wildlife, cats (Felis catus), and cows (Bos taurus) on a farm in Tennessee, U.S. in 2016 and 2017. Twenty-two soil samples were collected from the farm and subjected to oocyst flotation, DNA extraction, and polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) targeting 18S ribosomal RNA (18S rRNA) gene to detect and identify T. gondii. Three samples (13.6%) were positive for the parasite; however, T. gondii DNA was not consistently detected from repeated tests. Blood samples were collected from small mammals, cats, and mesopredators live-trapped on the farm, and serum from 30 of the farm's cows were obtained. Serological testing by the modified agglutination test (MAT; cutoff 1:50) found 2.5% (1/40) of small mammals, 52.9% (9/17) of raccoons (Procyon lotor), and 50% (1/2) of domestic cats were seropositive for T. gondii antibodies. No antibodies were found in 16 opossums (Didelphis virginiana), two skunks (Mephitis mephitis), and 30 cows. Small mammal tissue samples were subjected to PCR-RFLP detection. Four out of 29 (13.7%) tissue samples were positive for T. gondii; however, T. gondii DNA was not consistently detected during repeated PCR-RFLP testing. Our results indicate the ability to detect T. gondii varies greatly by contamination indicator. We found detection of soil oocysts to be challenging, and results suggest limited utility of the method performed. The ability to detect T. gondii in animals was highly variable among species. Our research emphasizes the importance of a holistic approach when surveilling for T. gondii to compensate for shortcomings of each contamination indicator. Future research should be conducted to further investigate the most effective T. gondii surveillance methods and species with increased sample sizes at other agricultural facilities.

Recent epidemiologic, clinical, subclinical and genetic diversity of Toxoplasma gondii infections in bats.

The protozoan Toxoplasma gondii infects virtually all warm-blooded animals, including bats. Depending on the diet, bats are classified as frugivorous, insectivorous, omnivorous, hematophagous, nectarivorous and carnivorous. The seroprevalence of T. gondii was higher in insectivores bats than fructivores bats. Owls, eagles, and cats can be predators of bats. Bats may be important in the epidemiology of T. gondii because they can be sentinels and can spread infection. Viable T. gondii has been isolated from brain, heart and pectoral muscle of bats. ToxoDB genotypes #9, #10, #6, #19, #69, #162 were identified from bats tissues. Genotypes #9 is prevalent in Asia. Genotypes #6 is widely distributed across Africa and Brazil. This result was matched with genotypes distribution from other hosts. The present review summarizes worldwide information on the seroprevalence, molecular epidemiology, isolation, genotypes and clinical cases of T. gondii infection in bats. Further studies are needed to verify the validity of serological and molecular tests, and the transmission routes of T. gondii infection in bats.

Toxoplasma gondii Strain and Dose Effects on Feed Conversion Rate, Body Weight, Serum Antibodies Response, and Systemic Distribution in Intraperitoneally Infected Domestic Turkey Poults.

Toxoplasmosis, caused by the protozoan parasite Toxoplasma gondii, is an important foodborne zoonosis affecting a wide range of hosts, including birds. This study investigated the seroconversion, feed conversion rate, weight gain, and parasite tissue tropism as a function of parasite dose and virulence in turkeys. Twenty-five 4-wk-old female domestic turkeys (Meleagris gallapavo) were intraperitoneally infected with two different strains and two doses (105 and 108 tachyzoites/ml) of T. gondii tachyzoites, resulting in four treatment groups. A fifth group of 10 additional birds was intraperitoneally injected with sterile phosphate-buffered saline as a negative control. All birds remained subclinical except for three birds in the two high-dose groups (108 tachyzoites/ml). Survival rate was 88% (22/25). A 92% seroconversion rate was detected in T. gondii-infected birds using a modified agglutination test. Antibody titers as well as weight gain were related to the dose and strain of T. gondii used. Feed conversion rate was higher in the high-dose groups compared with low-dose and control groups, while weight gain was significantly lower at 14 days postinfection in the group infected with 108 tachyzoites/ml of virulent T. gondii strain. Gross lesions were detected in the pancreas and lungs of only one bird, and histopathologic findings varied depending on strain and dose. The organs that most frequently contained T. gondii DNA as detected by quantitative PCR were the brain and the heart, followed by the bursa of Fabricius and the lungs. This study confirmed that turkeys can be infected with T. gondii, and turkeys can show signs of infection when exposed to high doses. Given the increased practice of outdoor-raised livestock and wildlife consumption, continual experimental infection of T. gondii in wild and domestic animals should be pursued.

Artículo regular­Efectos de la cepa y la dosis de Toxoplasma gondii en la tasa de conversión alimenticia, el peso corporal, la respuesta de los anticuerpos séricos y la distribución sistémica en pavipollos domésticos infectados por vía intraperitoneal. La toxoplasmosis, causada por el parásito protozoario Toxoplasma gondii, es una zoonosis importante transmitida por los alimentos que afecta a una amplia gama de huéspedes, incluidas las aves. Este estudio investigó la seroconversión, la tasa de conversión alimenticia, el aumento de peso y el tropismo en los tejidos por el parásito en función de la dosis del parásito y su virulencia en pavos. Se infectaron intraperitonealmente veinticinco pavos domésticos hembras de 4 semanas (Meleagris gallapavo) con dos cepas diferentes y dos dosis (105 y 108 taquizoítos/ml) de taquizoítos de T. gondii, lo que resultó en cuatro grupos de tratamiento. Se inyectó intraperitonealmente un quinto grupo de 10 aves adicionales con solución salina amortiguada con fosfato estéril como control negativo. Todas las aves permanecieron subclínicas excepto tres aves en los dos grupos de dosis alta (108 taquizoítos/ml). La tasa de supervivencia fue del 88% (22/25). Se detectó una tasa de seroconversión del 92% en aves infectadas con T. gondii utilizando una prueba de aglutinación modificada. Los títulos de anticuerpos y el aumento de peso se relacionaron con la dosis y la cepa de T. gondii utilizada. La tasa de conversión alimenticia fue mayor en los grupos de dosis alta en comparación con los grupos de dosis baja y el control, mientras que el aumento de peso fue significativamente menor a los 14 días después de la infección en el grupo infectado con 108 taquizoítos/ml de cepa virulenta de T. gondii. Se detectaron lesiones macroscópicas en el páncreas y los pulmones de una sola ave y los hallazgos histopatológicos variaron según la cepa y la dosis. Los órganos que con mayor frecuencia contenían ADN de T. gondii detectado por PCR cuantitativa fueron el cerebro y el corazón, seguidos de la bolsa de Fabricio y los pulmones. Este estudio confirmó que los pavos pueden infectarse con T. gondii y que los pavos pueden mostrar signos de infección cuando se exponen a dosis altas. Dada la práctica cada vez mayor de consumo de ganado y vida silvestre criados al aire libre, se debe continuar evaluando la infección experimental continua de T. gondii en animales silvestres y domésticos.

Recent aspects on epidemiology, clinical disease, and genetic diversity of Toxoplasma gondii infections in Australasian marsupials.

BACKGROUND: Toxoplasma gondii infections are common in humans and animals worldwide. Among all intermediate hosts of T. gondii, captive marsupials from Australia and New Zealand are highly susceptible to clinical toxoplasmosis. However, most free-range marsupials establish chronic T. gondii infection. Infected marsupial meat may serve as a source of T. gondii infection for humans. Differences in mortality patterns in different species of kangaroos and other marsupials are not fully understood. Lifestyle, habitat, and the genotype of T. gondii are predicted to be risk factors. For example, koalas are rarely exposed to T. gondii because they live on treetops whereas wallabies on land are frequently exposed to infection. METHODS: The present review summarizes worldwide information on the prevalence of clinical and subclinical infections, epidemiology, and genetic diversity of T. gondii infecting Australasian marsupials in their native habitat and among exported animals over the past decade. The role of genetic types of T. gondii and clinical disease is discussed. RESULTS: Fatal toxoplasmosis has been diagnosed in captive Australasian marsupials in Argentina, Chile, China, Germany, Hungary, Japan, Spain, Turkey, and the USA. Most deaths occurred because of disseminated toxoplasmosis. Genetic characterization of T. gondii strains isolated from fatal marsupial infections identified Type III as well as atypical, nonclonal genotypes. Fatal toxoplasmosis was also diagnosed in free-ranging wombats (Vombatus ursinus) in Australia. Genetic characterization of DNA amplified directly from host tissues of subclinical culled kangaroos at slaughter identified many mixed-strain infections with both atypical and recombinant genotypes of T. gondii. CONCLUSIONS: Most Australasian marsupials in their native land, Australia and New Zealand, have high prevalence of T. gondii, and kangaroo meat can be a source of infection for humans if consumed uncooked/undercooked. The genotypes prevalent in kangaroos in Australia and New Zealand were genetically distinct from those isolated or genotyped from most macropods in the USA and other countries. Thus, clinical toxoplasmosis in marsupials imported from Australia is most likely to occur from infections acquired after importation.

Toxoplasma gondii prevalence in carnivorous wild birds in the eastern United States.

Toxoplasma gondii is an important zoonotic protozoan parasite that can infect all warm-blooded animals including mammals and birds. Raptors can be intermediate hosts for T. gondii and the infection may be dependent on their feeding habits. In this study, we investigated the seroprevalence of T. gondii in ten raptor species from Florida, Pennsylvania, and Tennessee followed by a parasite bioassay on select seropositive samples. From a total of 155 raptors, we detected T. gondii antibodies using a modified agglutination test (cutoff 1:25) in 32 (20.6%) birds. The T. gondii seroprevalence was 44.8% in Falconiformes (13/29), 75% in Strigiformes (15/20), and 3.8% in Ciconiiformes (4/106). All Ciconiiformes samples (hearts and sera) were collected from Pennsylvania during nuisance wildlife removal projects and all birds were apparently healthy. Falconiform and Strigiform samples were collected from an exotics clinic in Tennessee and a rehabilitation center in Florida. All sampled birds were dead or euthanatized due to failure of rehabilitation or treatment. There was no statistically significant difference in T. gondii seroprevalence between Tennessee and Florida in the tested raptors. There was also no statistically significant difference in T. gondii exposure between males and females or adults and subadults. Mice bioassay attempts using fresh brain and/or heart tissue were performed on four seropositive birds. We isolated viable T. gondii tachyzoites from one red-shouldered hawk (Buteo lineatus) and genotyped the isolate using polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) of ten genetic markers. The isolated strain was designated as TgHawkFL1, which is ToxoDB PCR-RFLP genotype #28. Further research is needed to investigate the prevalence of T. gondii in raptors in the United States to obtain a better understanding of the life cycle, wildlife population impacts, and transmission dynamics of the parasite.

Recent epidemiologic, clinical, and genetic diversity of Toxoplasma gondii infections in non-human primates.

Toxoplasma gondii infections are common in humans and animals worldwide. The present review summarizes worldwide information on the prevalence of clinical and subclinical infections, epidemiology, diagnosis, and genetic diversity of T. gondii in non-human primates (NHP) for the past decade. Seroprevalence estimates of T. gondii worldwide were tabulated for each host. Risk factors associated with T. gondii infections are evaluated. New World NHP in captivity are highly susceptible to T. gondii infection with high mortality associated with disseminated toxoplasmosis. T. gondii can be transmitted to NHP in contact with symptomatic NHP. Therefore, precautions should be taken to prevent transmission of T. gondii to humans while handling symptomatic NHP. There were no reports of clinical toxoplasmosis in Old World NHP. Among the different genera of New World NHP, susceptibility to clinical toxoplasmosis varies a great deal; however, factors affecting this susceptibility are not fully understood. Genetic characteristics of T. gondii strains from monkeys is summarized.