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Toxoplasmosis has been recognized as a major public health problem worldwide. The consumption of uncooked or undercooked meat infected with Toxoplasma gondii tissue cysts is one of the main means of transmission of this parasite. Although sheep, goats, and pigs are commonly infected with T. gondii, little information is available on the distribution of T. gondii tissue cysts in naturally infected meat. In this study, we investigated the distribution of viable T. gondii tissue cysts in shoulder muscles of naturally infected lambs and goats. Hearts and shoulders of 46 lambs and 39 goats from a local grocery store were tested for T. gondii infection. Animals were evaluated for the presence of anti–T. gondii antibodies in heart blood and clots by the modified agglutination test. Fourteen of the 85 animals (seven lambs and seven goats) were seropositive. Six to 12 samples weighing 5, 10, and 50 g were obtained from shoulder muscles of each seropositive animal and used for bioassay in mice. The distribution of viable T. gondii differed according to the size of the sample analyzed, but in general larger sample sizes resulted in higher isolation rates (P , 0.05). Results of the study revealed an uneven distribution of T. gondii in muscle samples of lambs and goats and that T. gondii can be transmitted by consumption of very small servings (5 and 10 g) of meat when it is consumed raw or is undercooked. [ABSTRACT FROM AUTHOR]

Abstract: Hepatozoon sp. stages were detected in histological sections of the muscles of 11 of 25 bobcats (Lynx rufus). Parasites were detected from the myocardium of 11, tongues of 4, and limb muscles of 5 of these animals, which had been hunted in Mississippi in 2017. The intensity of infection was highest in the heart. Only asexual stages (meronts) were detected. Three types of meronts (types 1, 2, and 3) were detected, based on structure and mode of division. Additionally, individual zoites were found in leukocytes in the blood vessels of the myocardium, but the stage was not identified. Based on genomic DNA characterized from paraffin-embedded myocardium sections from 1 bobcat using 18S rRNA gene, the Hepatozoon species from the bobcat was related to but distinct from other Hepatozoon spp. of felids. This is the first description of the development of Hepatozoon in the muscles of bobcats. A new name, Hepatozoon rufi, is proposed for this parasite in bobcats., (© American Society of Parasitologists 2024.)

Trichinella infections have been eliminated from pork where pigs are raised in biosecure facilities, but wildlife infections persist. Trichinella murrelli is the primary zoonotic species in wild carnivores in the United States, having been identified in several species of omnivores and carnivores. Here, we document its occurrence in seven of 21 (33.3%) red foxes (Vulpes vulpes) from six counties in Pennsylvania. Encysted Trichinella larvae were detected in muscle squashes (<5 g samples) of all seven foxes, and in histological sections of the tongue and limb muscle of three. Larvae from muscle squashes were pooled and tested in a multiplex PCR capable of differentiating all Trichinella species native to the USA; all samples contained only T. murrelli. This is the first identification of T. murrelli in red foxes from Pennsylvania, and the first such survey performed in the last three decades. Results indicate that Trichinella remains endemic in Pennsylvania wildlife and a threat to the health of those who consume wild game., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024. Published by Elsevier B.V.)

Muscles of 25 bobcats (Lynx rufus) from remote areas of Mississippi in 2017 were tested for parasites. Testing for Sarcocystis infections included microscopic examination of fresh unstained muscle squashes, pepsin digestion of hearts and tongues, and histological sections of paraffin-embedded tissues. Sarcocystis spp. infections were detected in the muscles of 21 (84%) by a combination of methods. Sarcocysts were detected in the unstained tongue squashes of 2 bobcats. Sarcocystis sp. bradyzoites were detected in the pepsin digests of 3 of 19 hearts, and 12 of 19 tongues. In paraffin-embedded histological sections, sarcocysts were detected in 7 of 25 hearts, 17 of 25 tongues, and 5 of 23 limb muscles. Based on the character of the cyst wall, at least 3 morphologic types of sarcocysts were detected: those with small spikes on the cyst wall, corresponding to Sarcocystis felis, those with long villar protrusions, corresponding to Sarcocystis neurona, and those lacking visible cyst wall protrusions, representing an unidentified type of sarcocyst. Myositis associated with sarcocysts was seen in the tongues of 3, and in the limb muscles of 1 bobcat. Multilocus genotyping of the DNA extracted from paraffin-embedded sections from 2 bobcats, employing 18S, 28S, COI, ITS-1, and 5.8S and rpoB genes, diagnosed Sarcocystis caninum, S. felis, Sarcocystis lutrae, and S. neurona. An encapsulated species of Trichinella was identified in the tongue of 1; it represents the first documented occurrences in bobcats from Mississippi. Taken together, these observations suggest intensive exposure of these wild carnivores to Trichinella tissue cysts, implies predation or scavenging on these tissues promotes parasite transmission, and raises caution concerning zoonotic risk when such meat is rendered for human consumption., (© American Society of Parasitologists 2024.)

Here, we report the first known outbreak of clinical protozoal myeloencephalitis in naturally infected raccoons by the parasite Sarcocystis neurona. The North American opossum (Didelphis virginiana) and the South American opossum (Didelphis albiventris) are its known definitive hosts. Several other animal species are its intermediate or aberrant hosts. The raccoon (Procyon lotor) is considered the most important intermediate host for S. neurona in the USA. More than 50% of raccoons in the USA have sarcocysts in their muscles, however clinical sarcocystosis in raccoons is rare. In 2014, 38 free-living raccoons were found dead or moribund on the grounds of the Saint Louis Zoo, Missouri, USA. Moribund individuals were weak, lethargic, and mildly ataxic; several with oculo-nasal discharge. Seven raccoons were found dead and 31 were humanely euthanized. Postmortem examinations were conducted on nine raccoons. Neural lesions compatible with acute sarcocystosis were detected in eight raccoons. The predominant lesions were meningoencephalitis and perivascular mononuclear cells. Histologic evidence for the Canine Distemper Virus was found in one raccoon. Schizonts and merozoites were present in the encephalitic lesions of four raccoons. Mature sarcocysts were present within myocytes of five raccoons. In six raccoons, S. neurona schizonts and merozoites were confirmed by immunohistochemical staining with S. neurona-specific polyclonal antibodies. Viable S. neurona was isolated from the brains of two raccoons by bioassay in interferon gamma gene knockout mice and in cell cultures seeded directly with raccoon brain homogenate. Molecular characterization was based on raccoon no. 68. Molecular characterization based on multi-locus typing at five surface antigens (SnSAG1-5-6, SnSAG3 and SnSAG4) and the ITS-1 marker within the ssrRNA locus, using DNA isolated from bradyzoites released from sarcocysts in a naturally infected raccoon (no. 68), confirmed the presence of S. neurona antigen type I, the same genotype that caused a mass mortality event in which 40 southern sea otters stranded dead or dying within a 3 week period in April 2004 with S. neurona-associated disease. An expanded set of genotyping markers was next applied. This study reports the following new genotyping markers at 18S rRNA, 28S rRNA, COX1, ITS-1, RON1, RON2, GAPDH1, ROP20, SAG2, SnSRS21 and TUBA1 markers. The identity of Sarcocystis spp. infecting raccoons is discussed., (Published by Elsevier Ltd.)

Consumption of undercooked meat is one of the main transmission routes for Toxoplasma gondii worldwide. In the South American Andes, the guinea pig (Cavia porcellus) is a domestic rodent representing one of the main sources of animal proteins for indigenous communities. Although T. gondii infects a wide range of rodents worldwide, the natural impact of the infection on guinea pig populations is still unknown. Our study conducted in guinea pigs that were bred in traditional systems located in the village of José María Hernández (Nariño, Colombia) revealed the presence of T. gondii antibodies in 33.3% (23 out of 69) guinea pigs evaluated, with a cut-off point of 25 for the modified direct agglutination test. Conventional PCR detection of the T. gondii-specific RE fragment (529 bp) in 207 collected tissues demonstrated the presence of T. gondii DNA in several organs, including the brain (16/69), muscle (12/69), and heart (4/69), with an overall molecular detection frequency of 27.5% (19 out of 69 guinea pigs). This is the first report of natural infection of guinea pigs with T. gondii, demonstrating their potential epidemiological role in transmitting the infection to autochthonous populations., Competing Interests: Declaration of Competing Interest The authors declare no conflict of interest., (Copyright © 2022 The Authors. Published by Elsevier B.V. All rights reserved.)

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., (Copyright © 2021 Elsevier Ltd. All rights reserved.)

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., (Published by Elsevier Ltd.)

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 wild canids and other carnivores for the past decade. Seroprevalence estimates of T. gondii worldwide were tabulated for each host. Seroprevalence in wild foxes was very high compared with farmed Arctic foxes. Economic and public health aspects of some of the carnivore species raised for fur and meat (raccoon dogs, mink) are discussed. Diagnostic efficacies of different serological methods and PCR methods are discussed. Clinical toxoplasmosis was observed mainly in carnivores concurrently infected with immunosuppressive Canine Distemper Virus infection. Abortion and blindness were noted in mink. Genetic diversity of isolates using DNA derived from 162 (89 viable T. gondii isolates and 73 DNA extracted from tissues) of wild carnivores from several countries is discussed. However, 69 of the 162 T. gondii isolates were strains from USA and these were genetically diverse with predominance of ToxoDB genotypes #4 and #5 (haplogroup 12). Only limited information is available concerning genotyping of T. gondii isolates from other countries; none of the 93 T. gondii isolates from other countries (Brazil, China, France, Grenada) were haplogroup 12., (Published by Elsevier B.V.)

Toxoplasma gondii infections are common in humans and animals worldwide. T. gondii causes mortality in several species of marine mammals, including threatened Southern sea otters (Enhydra lutris) and endangered Hawaiian monk seals (Monachus schauinslandi). Marine mammals are now considered sentinels for environmental exposure to protozoan agents contaminating marine waters, including T. gondii oocysts. Marine mammals also serve as food for humans and can result in foodborne T. gondii infections in humans. The present review summarizes worldwide information on the prevalence of clinical and subclinical infections, epidemiology, and genetic diversity of T. gondii infecting marine mammals in the past decade. The role of genetic types of T. gondii and clinical disease is discussed., (Published by Elsevier B.V.)

Toxoplasma gondii infections are common in humans and animals worldwide. Toxoplasma gondii infection in pigs continues to be of public health concern. Pigs are important for the economy of many countries, particularly, USA, China, and European countries. Among the many food animals, pigs are considered the most important for T. gondii transmission in USA and China because viable parasites have rarely been isolated from beef or indoor raised chickens. Besides public health issues, T. gondii causes outbreaks of clinical toxoplasmosis in pigs in China, associated with a unique genotype of T. gondii (ToxoDB genotype #9 or Chinese 1), rarely found in other countries. The safety of ready to eat pork products with respect to T. gondii infection is a matter of recent debate. Here, we review in detail seroprevalence, prevalence of viable and nonviable T. gondii, epidemiology, risk assessment, diagnosis, and curing of pork products containing T. gondii for the past decade. This review will be of interest to biologists, parasitologists, veterinarians, and public health workers., (Published by Elsevier B.V.)

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 dogs (Canis familiaris) from 2009-2020. Seroprevalence estimates of T. gondii worldwide were tabulated. Reports of high seroprevalence in canine population and high congenital transmission of T. gondii in dogs in Brazil are reviewed. Most reports from China were published in Chinese, and these reports are now summarized here. Dogs have an additional importance in some countries such as China, Vietnam, and Nigeria; whereas in many cities dog meat is sold commercially for human consumption and given to felids, and transmission of T. gondii could occur if meat is not cooked properly. Dogs can ingest T. gondii-infected cat feces and these oocysts remain viable after passage through the digestive tract of the dog; T. gondii DNA was found in feces of dogs from New York City parks in USA. Most clinical canine cases of toxoplasmosis were in immunosuppressed dogs, and ulcerative dermatitis was one of the main presentations. Genetic diversity based on PCR-RFLP markers using DNA derived from 133 viable T. gondii isolates from dogs from several countries is discussed. T. gondii strains from Asia and Americas were more genetically diverse than those from Africa. This review will be of interest to biologists, parasitologists, veterinarians, and public health workers., (Published by Elsevier B.V.)

In a national survey of fresh, unfrozen, American pasture-raised lamb and pork, the prevalence of viable Toxoplasma gondii was determined in 1500 samples selected by random multistage sampling (750 pork, 750 lamb) obtained from 250 retail meat stores from 10 major geographic areas in the USA. Each sample consisted of a minimum of 500g of meat purchased from the retail meat case. To detect viable T. gondii , 50g meat samples of each of 1500 samples were bioassayed in mice. Viable T. gondii was isolated from 2 of 750 lamb samples (unweighted: 0.19%, 0.00-0.46%; weighted: 0.04%, 0.00-0.11%) and 1 of 750 pork samples (unweighted: 0.12%, 0.00-0.37%; weighted: 0.18%, 0.00-0.53%) samples. Overall, the prevalence of viable T. gondii in these retail meats was very low. Nevertheless, consumers, especially pregnant women, should be aware that they can acquire T. gondii infection from ingestion of undercooked meat. Cooking meat to an internal temperature of 66°C kills T. gondii ., Competing Interests: Conflict of interest None.

The importance of birds in the biological cycle of Neospora caninum is not clear. We report unsuccessful Neospora infection in chickens (Gallus gallus domesticus) using two isolates of N. caninum. In experiment #1, 30 White Leghorn chickens were orally inoculated with viable N. caninum oocysts (NC-SP1 isolate, 200 oocysts per bird) via the crop at 21days of age. Groups of three birds were euthanised at intervals of 7days (a total of 9weeks) and one group was challenged with the same oocyst dose at 37daysp.i. and observed for 11weeks. Blood samples were collected weekly, and sera were tested using IFAT. Chicken tissues were collected for PCR, quantitative PCR and immunohistochemistry. Two dogs approximately 45days of age were fed with tissues from chickens euthanised at 138 and 159daysp.i. The results indicated that the chickens were resistant to neosporosis as revealed by failure to seroconvert, to detect parasite DNA or N. caninum antigen by immunohistochemistry in inoculated bird tissues, and by no oocyst excretion by the dogs fed avian tissues. Similar results were obtained in experiment #2, in which 34 1-week-old chickens were each s.c. inoculated with 100,000 tachyzoites of the NcWTDMn1 isolate of N. caninum. The chickens were euthanised on days 7, 15, 22, 28, 36 and 60p.i. At necropsy, all tissues and serum from each bird were collected. All chickens remained asymptomatic, and N. caninum antigen was not detected by immunohistochemistry. Seven chickens euthanised at day 60p.i. demonstrated low (1:25 dilution) levels of antibodies by using the Neospora agglutination test. Two 12-week-old dogs fed tissues pooled from 10 inoculated chickens euthanised at day 60p.i. did not excrete N. caninum oocysts. This investigation indicates that chickens are resistant to experimental infection by N. caninum., (Copyright © 2017 Australian Society for Parasitology. Published by Elsevier Ltd. All rights reserved.)

Chickens are considered important in the epidemiology of Toxoplasma gondii. Chicken hearts (n = 1185) obtained from grocery stores were tested for T. gondii infection. Antibodies to T. gondii were assayed in fluid removed from the heart cavity using the modified agglutination test (MAT) at 1:5, 1:25, and 1:100 dilutions. MAT antibodies were detected in 222 hearts at 1:5 dilution and 8 hearts at 1:25 dilution, but none were positive at 1:100 dilution. Seropositive (n = 230, 19.4%) chicken hearts were bioassayed in mice and seronegative (n = 157) chickens were bioassayed in cats. Viable T. gondii was not isolated from any hearts by bioassays in mice. The 2 cats fed 60 and 97 hearts did not excrete T. gondii oocysts. The results indicate a low prevalence of viable T. gondii in chickens from grocery stores. Molecular typing of 23 archived T. gondii strains isolated from free-range chickens from Ohio and Massachusetts using the 10 PCR-RFLP markers including SAG1, SAG2 (5'-3'SAG2 and altSAG2), SAG3, BTUB, GRA6, c22-8, c29-2, L358, PK1, and Apico revealed that seven were ToxoDB PCR-RFLP genotype #1, 11 were genotype #2, one was genotype #3, three were genotype #170, and one was mixed genotype. These results indicate that the clonal genotypes #1 (type II), #2 (type III), and #3 (type II variant) are common in free-range chickens.

The protozoan Toxoplasma gondii and the metazoan Trichinella spp. infect virtually all warm-blooded animals, including birds, humans, livestock, and marine mammals. Both parasitic infections can cause serious illness in human beings and can be acquired by ingesting under-cooked meat harboring infective stages. Approximately 3500 black bears (Ursus americanus) are legally-harvested each year in Pennsylvania, USA during the November hunting season. Among animals found infected with T. gondii, the prevalence of T. gondii is the highest among black bears in the USA; however, little is currently known of epidemiology of toxoplasmosis in this host species. Serum samples were collected during the winters of 2015 and 2016 from adult female bears and their nursing cubs or yearlings while they were still in their dens. Additionally, archived sera from bear samples collected throughout the year, including hunter-harvested bears in November and trapped bears in the summer, were serologically tested. Antibodies to T. gondii were assayed by the modified agglutination test (MAT, cut-off 1:25) and antibodies to Trichinella spp. were assayed using a commercial enzyme-linked immunosorbent assay (ELISA). Overall, T. gondii antibodies were found in 87.6% (206/235) of adults, and 44.1% (30/68) of yearlings. In March 2015/2016 sampling, antibodies to T. gondii were found in 94% (30/32) adult female bears while in their den. Antibodies were detected in 5% (3/66) of the nursing cubs in the dens of these sows. One positive cub had a MAT titer of 1:160 and two were positive at the 1:25 dilution but not at 1:50. The adult females of these cubs had MAT titers ranging from 1:400 to 1:3200. Antibodies to Trichinella spp. were found in 3% (6/181) of adults and 3.6% (1/28) of yearlings; these 7 bears were also seropositive for T. gondii. No antibodies to Trichinella spp. were detected in the sera of 44 nursing cubs tested. The finding of T. gondii antibodies in only 3 of 66 cubs, and higher antibody titers in their respective sows indicates that the colostrally-acquired antibodies wane to undetectable levels by 8-10 weeks, while the cubs are still in the den. The results indicate that there is no transplacental transmission of T. gondii, that antibodies acquired from colostrum are largely undetectable by the time cubs emerge from the den, and nearly that 50% of bears acquire infection postnatally by 10 months of age. This is the first report of disappearance of transcolostral antibodies of any infection in bears., (Copyright © 2016. Published by Elsevier B.V.)

Little is known of the epidemiology of toxoplasmosis in Minnesota. Here, we evaluated Toxoplasma gondii infection in 50 wild bobcats (Lynx rufus) and 75 other animals on/near 10 cattle farms. Antibodies to T. gondii were assayed in serum samples or tissue fluids by the modified agglutination test (MAT, cut-off 1:25). Twenty nine of 50 bobcats and 15 of 41 wildlife trapped on the vicinity of 10 farms and nine of 16 adult domestic cats (Felis catus) and six of 14 domestic dogs resident on farms were seropositive. Toxoplasma gondii oocysts were not found in feces of any felid. Tissues of all seropositive wild animals trapped on the farm were bioassayed in mice and viable T. gondii was isolated from two badgers (Taxidea taxus), two raccoons (Procyon lotor), one coyote (Canis latrans), and one opossum (Didelphis virginiana). All six T. gondii isolates were further propagated in cell culture. Multi-locus PCR-RFLP genotyping using 10 markers (SAG1, SAG2 (5'-3'SAG2, and alt.SAG2), SAG3, BTUB, GRA6, c22-8, c29-2, L358, PK1, and Apico), and DNA from cell culture derived tachyzoites revealed three genotypes; #5 ToxoDataBase (1 coyote, 1 raccoon), #1 (1 badger, 1 raccoon, 1 opossum), and #2 (1 badger). This is the first report of T. gondii prevalence in domestic cats and in bobcats from Minnesota, and the first isolation of viable T. gondii from badger., (Published 2016. This article is a U.S. Government work and is in the public domain in the USA.)