Isolation and Genetic Characterization of Toxoplasma gondii from a Patas Monkey (Erythrocebus patas) in China.

Many cases of Toxoplasma gondii infection have been reported worldwide in non-human primates (NHPs), especially in captive New World monkeys. However, few studies on toxoplasmosis in Old World monkeys have been conducted. In this study, serological and molecular biological analyses were carried out to look for T. gondii antibodies and T. gondii infection in 13 NHPs from China. T. gondii infection was confirmed in 8 NHP cases. T. gondii antibodies were detected in 1/5 New World monkeys and in 4/7 Old World monkeys. T. gondii DNA was detected in 3/5 New World monkeys and 5/7 Old World monkeys. The one ring-tailed lemur was negative for both antibodies and DNA of T. gondii. The most common clinical manifestations of T. gondii infection were malaise, poor appetite, emaciation, and foamy nasal discharge. The most common histopathological findings were interstitial pneumonia, necrotic hepatitis, necrotizing myocarditis, lymphadenitis, and necrotic splenitis. One viable T. gondii strain was successfully isolated from the myocardium of a patas monkey (Erythrocebus patas) by bioassay in mice. T. gondii tachyzoites were obtained from cell cultures and were designated as TgMonkeyCHn2. The genotype of this strain belongs to ToxoDB genotype #9, and the allele of ROP18/ROP5 gene was 3/6. TgMonkeyCHn2 tachyzoites were avirulent in Swiss mice. To our knowledge, this is the first report of fatal toxoplasmosis in a patas monkey. T. gondii infection in patas monkeys may indicate environmental contamination by oocysts. The patas monkey is a new host record for T. gondii.

Low Prevalence of Toxoplasma gondii in Sheep and Isolation of a Viable Strain from Edible Mutton from Central China.

Sheep are highly susceptible to Toxoplasma gondii, and miscarriage is the main clinical feature. This study investigated 227 sheep samples (210 myocardial tissues from slaughterhouses, 6 ewe serum samples, 3 aborted fetuses, and 8 dead lambs from veterinary clinics) from central China for T. gondii infection. Antibodies against T. gondii were detected using the modified agglutination test (MAT). PCR was performed to detect T. gondii DNA in the tissue samples. The results showed that four samples were seropositive (MAT titer ≥ 1:100), with a seroprevalence of 1.8% (4/227). The seropositive samples included two myocardial samples from a slaughterhouse, one ewe and its aborted fetus from a veterinary clinic. The results revealed that 7 out of 207 (3.4%) sheep tissue samples were PCR-positive, including two myocardial tissue samples from slaughterhouses, three aborted fetuses, and two lambs from veterinary clinics. Toxoplasma gondii vertical transmission had occurred in two of three pairs of ewes and her pups. One viable T. gondii strain (TgSheepCHn14) was isolated from the myocardial tissues of sheep from a slaughterhouse. Tachyzoites were obtained from cell cultures at 70 days following seeding in the brains and lungs of mice. This strain was non-lethal to Swiss mice. The number of parasite brain cysts in mice decreased with time post-infection (p < 0.05). Overall, the prevalence of T. gondii in the sheep samples was low. Although the samples were scattered, and not from planned collections, the current study detected T. gondii antibodies and DNA in aborted fetuses, indicating that vertical transmission could occur and maintain the parasites in sheep herds without exogenous infection.

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.

Low Prevalence of Toxoplasma gondii in Dogs From Central China.

Background: Toxoplasma gondii can infect almost all warm-blooded animals, including humans and dogs. Humans can become infected with T. gondii by petting dogs that have eaten or contacted infected cat feces. The aim of this study was to evaluate T. gondii infections in dogs from central China. From 2015 to 2021, a total of 536 dog samples (195 fecal, 81 hearts, and 260 serum samples) from Henan Province were collected. Heart juice or serum samples (n = 341) were tested for T. gondii antibodies using the modified agglutination test (MAT). Fresh myocardium (n = 6) and blood (n = 2) samples were bioassayed in mice. Results: The present study showed that 4.40% (15/341) of the dogs were seropositive for T. gondii by MAT (cut-off, 1:25) and 4.10% (8/195) of dog feces contained T. gondii DNA. No T. gondii DNA was found in any myocardium (n = 81) or blood (n = 2) samples. The viable T. gondii strain was not isolated from any myocardium or blood samples (n = 8). Compared to the prevalence of T. gondii antibodies in dogs sampled from 2015 to 2018, the prevalence significantly declined from 2020 to 2021 (P < 0.05). Gender and age were not risk factors for dogs infected with T. gondii in this study. However, compared to other sources, dogs from Zhoukou City (close to the Yellow River) or from pet shops showed significantly higher prevalence for T. gondii (P < 0.05). Conclusion: A total of 4.29% dogs were infected by T. gondii (23/536, 8 of 195 fecal samples, 2 of 260 serum, and 13 of 81 heart juice samples). This is the first survey of T. gondii infection in dog feces from China. Dogs were exposed to T. gondii, and they could act as mechanical transmitters of T. gondii.

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.

First Report of Sarcocystis Masoni in a Captive Alpaca (Vicugna Pacos) From China.

Background: Sarcocystosis is a parasitic disease caused by intracellular protozoan parasite of the genus Sarcocystis. Tissue samples of alpacas (n = 4) from Henan province (China) were screened for Sarcocystis spp. infection by histological examination, pepsin digestion, and molecular assays. Results: Sarcocystis spp. was detected in heart, liver, spleen, lung, and kidney of an alpaca by molecular assays. Many sarcocysts with inflammation responses were observed in this alpaca myocardium, and they showed a high similarity to Sarcocystis masoni by sequence analysis. Conclusion: This study is the first to demonstrate Sarcocystis spp. infection in alpaca from China. The higher parasite load in the alpaca myocardium indicated that it had contact with an environment contaminated with sporocysts, and that the alpaca was susceptible to Sarcocystis spp.

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.

Lower seroprevalence of Toxoplasma gondii in swine from central China after an outbreak of African swine fever.

Toxoplasma gondii is widespread worldwide and can infect swine. This study evaluated the seroprevalence of T. gondii in swine from central China after an outbreak of African swine fever (ASF). A total of 2683 swine serum samples were collected from farms in four provinces. Of the serum samples, 1.42% (38/2683) (95% CI, 1.03-1.94) tested positive for T. gondii IgG antibody by a modified agglutination test (MAT) (cut-off: 1:25). Comparing with the results of previous studies, specifically our survey from before the outbreak, the seroprevalence of T. gondii in swine from central China was significantly decreased after the occurrence of ASF (OR = 7.679, 2015-2017 vs. 2019-2020). In general, the proportion of seropositive animals increased with the age of the swine, indicating post-natal transmission of T. gondii. Furthermore, there was a significant difference in seroprevalence between suckling pigs and weaned pigs (p < 0.05). This is the first large-scale investigation of T. gondii infection in swine after an ASF outbreak in China. The lower seroprevalence of T. gondii in swine after ASF may be due to stricter biosecurity measures on the farms, but results indicated swine exposure to zoonotic parasites despite these measures. This highlights that pigs must be considered a potential source of human T. gondii infections.

TITLE: Baisse de la séroprévalence de Toxoplasma gondii chez les porcs du centre de la Chine après une épidémie de peste porcine africaine. ABSTRACT: Toxoplasma gondii est répandu dans le monde entier et peut infecter les porcs. Cette étude a évalué la séroprévalence de T. gondii chez les porcs du centre de la Chine après une épidémie de peste porcine africaine (PPA). Au total, 2 683 échantillons de sérum de porc ont été prélevés dans des fermes de quatre provinces. Parmi les échantillons de sérum, 1,42 % (38/2683) (IC à 95 %, 1,03­1,94) étaient positifs pour les anticorps IgG de T. gondii par un test d'agglutination modifié (MAT) (seuil : 1:25). En comparaison avec les résultats d'études précédentes, en particulier notre enquête d'avant l'épidémie, la séroprévalence de T. gondii chez les porcs du centre de la Chine a été significativement diminuée après l'apparition de la PPA (OR = 7,679, 2015-2017 vs 2019-2020). En général, la proportion d'animaux séropositifs augmentait avec l'âge des porcs, indiquant une transmission postnatale de T. gondii. De plus, il y avait une différence significative entre la séroprévalence chez les cochons de lait et les cochons sevrés (p < 0,05). Il s'agit de la première enquête à grande échelle sur l'infection à T. gondii chez le porc après l'épidémie de PPA en Chine. La séroprévalence plus faible de T. gondii chez les porcs après la PPA peut être due à des mesures de biosécurité plus strictes dans les fermes, mais les résultats ont indiqué une exposition des porcs aux parasites zoonotiques malgré ces mesures. Cela souligne que les porcs doivent être constamment considérés comme une source potentielle d'infection humaine à T. gondii.

Comparative Immunological Response and Pathobiology of Mice Inoculated with Toxoplasma gondii Isolated from Different Hosts.

Toxoplasma gondii is an obligate intracellular parasite that has a worldwide distribution and can infect almost all warm-blood animals. Serological tests are the main detection methods for T. gondii infection in animals and humans. Little is known of biological behavior, antibody responses, and virulence of T. gondii strains in mice from China. Here we document antibody responses, tissue cyst burden, and mouse virulence of T. gondii strains isolated from different hosts in China. All T. gondii strains formed tissue cysts in the brains of mice and positively correlated with the T. gondii antibody titer (R2 = 0.3345). These results should aid in the diagnosis and characterization of T. gondii isolates.

Toxoplasma gondii infection in white spoonbills (Platalea leucorodia) from Henan Province, China.

Toxoplasma gondii oocysts in the environment are a threat to humans and animals. This study is aimed to evaluate the prevalence of T. gondii in white spoonbills and isolate viable T. gondii from white spoonbills. In 28.6% (2/7) of white spoonbills, T. gondii antibodies were found in heart juice by the modified agglutination test (cut-off: 1:4). T. gondii DNA was detected in tissues of 42.9% (3/7) white spoonbills. One viable T. gondii strain, named TgSpoonbillCHn1, was isolated from the myocardium of a white spoonbill by bioassay in mice. DNA extracted from TgSpoonbillCHn1 tachyzoites was characterized by PCR-restriction fragment length polymorphism with ten markers and the virulence genes ROP5 and ROP18. The results revealed that it was ToxoDB#2 (Type III). The ROP18/ROP5 genotype combination predicts that this strain is avirulent for mice, which is supported by the infection experiments in mice. This is the first report of the isolation of viable T. gondii strain from white spoonbil. The prevalence of T. gondii in white spoonbills may be indicative of environmental contamination of oocysts. This report provides direct evidence of white spoonbill as an intermediate host of T. gondii.

Isolation and characterization of Toxoplasma gondii from captive caracals (Caracal caracal).

Toxoplasma gondii infects most warm-blooded animals, including humans. Felids can serve as both intermediate and definitive hosts for T. gondii. However, there is no direct evidence to prove the caracal (Caracal caracal) is an intermediate host for T. gondii. Here, we report T. gondii infection in two caracals in a zoo from China. Antibodies against T. gondii were found in both caracals by modified agglutination test (MAT) (cut-off titer: 1:25). Tissue cysts were observed in the leg and tongue muscles of caracal case# 1. These cysts were confirmed as T. gondii by immunohistochemical staining and T. gondii was detected by polymerase chain reaction (PCR). Viable T. gondii strain was isolated from the striated muscles of caracal case# 2 and designated as TgCaracalCHn1. DNA from tachyzoites obtained from cell cultures was characterized by RFLP-PCR using ten markers (SAG1, SAG3, SAG2, BTUB, c22-8, GRA6, c29-2, PK1, L358, and Apico) and the virulence genes (ROP5 and ROP18). The results indicate that this isolate belongs to ToxoDB genotye #2 (Type III). The virulence of this isolate was evaluated in BALB/c mice. A dose of 104 TgCaracalCHn1 tachyzoites was non-lethal to mice. Tissue cysts were found in brain tissues of infected mice. This result confirmed that the TgCaracalCHn1 is non-virulent to mice. Current study documents first isolation of viable T. gondii strain from caracal and also indicates that caracal can act as new intermediate host for T. gondii.

Low Prevalence of Antibodies Against Toxoplasma gondii in Chinese Populations.

Toxoplasma gondii has been found to infect almost all warm-blooded animals, including humans. In this study, a total of 3,275 human serum samples were collected from hospitals in five provinces of China. About 5.13% (168/3,275) (95% CI, 4.42-5.94) of the serum samples tested positive for T. gondii IgG antibody by a modified agglutination test (MAT) (cut-off: 1:20). Significant associations were detected between geographic location (OR = 1.763), age (OR = 3.072), infertility in women (OR = 2.4409) and T. gondii infection in humans (p < 0.05). To minimize infection, citizens need to be informed about the best practices for toxoplasmosis prevention, including eating well-cooked meat, drinking boiled water, washing vegetables and fruits, and being careful during contact with cats.