ABORTION AND NEONATAL MORTALITY DUE TO TOXOPLASMA GONDII IN BIGHORN SHEEP (OVIS CANADENSIS).

Low lamb recruitment can be an obstacle to bighorn sheep (Ovis canadensis) conservation and restoration. Causes of abortion and neonate loss in bighorn sheep, which may affect recruitment, are poorly understood. Toxoplasma gondii is a major cause of abortion and stillbirth in domestic small ruminants worldwide, but no reports exist documenting abortion or neonatal death in bighorn sheep attributable to toxoplasmosis. Between March 2019 and May 2021, eight fetal and neonatal bighorn lamb cadavers from four western US states (Idaho, Montana, Nebraska, and Washington) were submitted to the Washington Animal Disease Diagnostic Laboratory for postmortem examination, histologic examination, and ancillary testing to determine the cause of abortion or neonatal death. Necrotizing encephalitis characteristic of toxoplasmosis was identified histologically in six of eight cases, and T. gondii infection was confirmed by PCR in five cases with characteristic lesions. Other lesions attributable to toxoplasmosis were pneumonia (3/5 cases) and myocarditis (2/5 cases). Protozoal cysts were identified histologically within brain, lung, heart, skeletal muscle, adipose tissue, or a combination of samples in all five sheep with PCR-confirmed T. gondii infections. Seroprevalence of T. gondii ranged from 40-81% of adult females sampled in the Washington population in October and November 2018-2021, confirming high rates of exposure before detection of Toxoplasma abortions in this study. Of 1,149 bighorn sheep postmortem samples submitted to Washington Animal Disease Diagnostic Laboratory between January 2000 and May 2021, 21 of which were from fetuses or neonates, a single case of chronic toxoplasmosis was diagnosed in one adult ewe. Recent identification of Toxoplasma abortions in bighorn sheep suggests that toxoplasmosis is an underappreciated cause of reproductive loss. Abortions and neonatal mortalities should be investigated through postmortem and histologic examination, particularly in herds that are chronically small, demographically stagnant, or exhibit reproductive rates lower than expected.

Prevalence and isolation of Toxoplasma gondii in goats slaughtered for human consumption in the semi-arid of northeastern Brazil.

The aim of this study was to determine the seroprevalence of anti-Toxoplasma gondii antibodies and factors associated with infection in goats, and to isolate protozoan strains in tissue samples from seropositive goats that were destined for human consumption in the state of Paraíba, northeastern Brazil. Serum samples from 229 slaughtered goats were tested using the indirect fluorescence antibody test (IFAT), with a cutoff point of 1:64. Epidemiological questionnaires were applied to the producers, to acquire information about the sanitary management used in their herds. Tissue samples from the animals were collected during slaughter, in order to perform bioassays in mice. The seroprevalence found was 21.39% (49/229), with antibody titers ranging from 1:64 to 1:32,768. The municipalities of origin, Patos (OR: 3.047; CI: 1.384-6.706) and Sousa (OR: 3.355; CI: 1.536-7.327), were considered to be factors associated with infection by T. gondii. Thirty-eight bioassays were performed in mice, using tissues from seropositive goats, with an isolation rate of 50% (19/38). There was no correlation between isolation rate and antibody titers. Only one mouse died, at 30 days post-infection, which demonstrated that the strains isolated had low virulence towards mice. It was concluded that there is high seroprevalence in goats in northeastern Brazil, as well as a high percentage of viable tissue cysts in slaughtered animals destined for human consumption. These results demonstrate that there is an imminent one health problem relating to toxoplasmosis, especially in the most populous municipalities in the study (Patos and Sousa), which were identified as factors associated with T. gondii infection in goats.

Acute exacerbation of pulmonary toxoplasmosis during corticosteroid therapy for immune thrombocytopenia: A case report and literature review.

RATIONALE: Pulmonary toxoplasmosis (PT) is an infectious disease that can be fatal if reactivation occurs in the recipients of hematopoietic stem cell transplantation (HSCT) who were previously infected with Toxoplasma gondii. However, whether the toxoplasmosis reactivation is an actual risk factor for patients receiving immunosuppressive therapies without HSCT remains unclear. Therefore, reactivated PT is not typically considered as a differential diagnosis for pneumonia other than in patients with HSCT or human immunodeficiency virus (HIV). PATIENT CONCERNS: A 77-year-old man presented with fever and nonproductive cough for several days. He was hospitalized due to atypical pneumonia that worsened immediately despite antibiotic therapy. Before 4 months, he was diagnosed with immune thrombocytopenia (ITP) and received corticosteroid therapy. Trimethoprim-sulfamethoxazole (ST) was administered to prevent pneumocystis pneumonia resulting from corticosteroid therapy. DIAGNOSIS: The serological and culture test results were negative for all pathogens except T. gondii immunoglobulin G antibody. Polymerase chain reaction, which can detect T. gondii from frozen bronchoalveolar lavage fluid, showed positive results. Therefore, he was diagnosed with PT. INTERVENTION: ST, clindamycin, and azithromycin were administered. Pyrimethamine and sulfadiazine could not be administered because his general condition significantly worsened at the time of polymerase chain reaction (PCR) examination. OUTCOMES: The patient died of acute respiratory distress syndrome despite anti-T. gondii treatment. An autopsy revealed a severe organizing pneumonia and a small area of bronchopneumonia. LESSONS: PT should be considered as a differential diagnosis in patients with pneumonia, particularly in seropositive patients who receive immunosuppressive therapies even for other than HSCT or HIV.

Presence of atypical genotypes of Toxoplasma gondii isolated from cats in the state of Bahia, Northeast of Brazil.

In this study, 20 blood, heart, and brain samples were collected from euthanized cats at the Zoonosis Control Centers and Veterinary Clinics in the state of Bahia, Brazil. The sera were examined for anti-T. gondii antibodies using the indirect hemagglutination test. The brains and hearts of seven seropositive cats were ground, and peptide digestion was performed for bioassay in mice. Toxoplasma gondii was isolated in 5/7 (71.42%) of seropositive cats. In these isolates, the parasite was genotyped using the Polymerase chain reaction, associated with the DNA fragment polymorphism obtained by restriction enzyme PCR-RFLP technique with 11 markers (SAG1, 5'-SAG2, 3'-SAG2, alt. SAG2, SAG3, BTUB, GRA6, c22-8, c29-2, L358, PK1, Apico, and CS3) and 15 microsatellite markers (TUB-2, W35, TgM-A, B18, B17, M33, IV.1, XI.1, M48, M102, N60, N82, AA, N61, N83). The analysis of the isolates by PCR-RFLP revealed five distinct genotypes. Three of these genotypes have never been reported before; one corresponded to the TgDgCo13 genotype, and one incomplete genotype. In genotyping analysis using microsatellite markers, it was observed that the isolates showed atypical alleles in the typing and fingerprint markers. This revealed five atypical genotypes. The typing marker B17 showed the highest degree of atypia. This study is the first to report the genotyping of T. gondii obtained from naturally infected cats in Bahia, Northeast Brazil. The genotypes found in this study were different from those found in other studies conducted in Bahia, which included different species of animals. None of the clonal lineages I, II, or III were found. This study demonstrates the diversity of T. gondii in the study region, with the presence of unusual genotypes, reaffirming the genetic variability of the parasite in Brazil.

Comparison between Enzyme Immunoassays Performed on Samples of Dried Blood and Serum for Toxoplasmosis Prenatal Screening: Population-based Study.

OBJECTIVE: Most prenatal screening programs for toxoplasmosis use immunoassays in serum samples of pregnant women. Few studies assess the accuracy of screening tests in dried blood spots, which are of easy collection, storage, and transportation. The goals of the present study are to determine the performance and evaluate the agreement between an immunoassay of dried blood spots and a reference test in the serum of pregnant women from a population-based prenatal screening program for toxoplasmosis in Brazil. METHODS: A cross-sectional study was performed to compare the immunoassays Imunoscreen Toxoplasmose IgM and Imunoscreen Toxoplasmose IgG (Mbiolog Diagnósticos, Ltda., Contagem, Minas Gerais, Brazil)in dried blood spots with the enzyme-linked fluorescent assay (ELFA, BioMérieux S.A., Lyon, France) reference standard in the serum of pregnant women from Minas Gerais Congenital Toxoplasmosis Control Program. RESULTS: The dried blood spot test was able to discriminate positive and negative results of pregnant women when compared with the reference test, with an accuracy of 98.2% for immunoglobulin G (IgG), and of 95.8% for immunoglobulin M (IgM). CONCLUSION: Dried blood samples are easy to collect, store, and transport, and they have a good performance, making this a promising method for prenatal toxoplasmosis screening programs in countries with continental dimensions, limited resources, and a high prevalence of toxoplasmosis, as is the case of Brazil.

OBJETIVO: A maioria dos programas de triagem pré-natal para toxoplasmose utiliza imunoensaios em amostras de soro de gestantes. Poucos estudos avaliam a acurácia dos testes de triagem em amostras de sangue seco, que são de fácil coleta, armazenamento e transporte. Este estudo teve como objetivo determinar o desempenho e avaliar a concordância entre um imunoensaio em sangue seco e um teste de referência em soro de gestantes de um programa de rastreamento pré-natal de base populacional para toxoplasmose no Brasil. MéTODOS: Realizou-se um estudo transversal para comparar os imunoensaios Imunoscreen Toxoplasmose IgM e Imunoscreen Toxoplasmose IgG (Mbiolog Diagnósticos, Ltda., Contagem, Minas Gerais, Brazil) em sangue seco com o padrão de referência ensaio fluorescente ligado a enzimas (enzyme-linked fluorescent assay, ELFA, BioMérieux S.A., Lion, França) no soro de gestantes do Programa de Controle de Toxoplasmose Congênita de Minas Gerais. RESULTADOS: O exame em sangue seco foi capaz de discriminar os resultados positivos e negativos das gestantes quando comparado ao teste de referência, com acurácia de 98,2% para imunoglobulina G (IgG), e de 95,8% para imunoglobulina M (IgM). CONCLUSãO: O sangue seco apresenta bom desempenho e é uma amostra de fácil coleta, armazenamento e transporte, o que o torna um método promissor para programas de triagem pré-natal de toxoplasmose em países com dimensões continentais, recursos limitados, e alta prevalência de toxoplasmose, como é o caso do Brasil.

Quantification of viable protozoan parasites on leafy greens using molecular methods.

Protozoan contamination in produce is of growing importance due to their capacity to cause illnesses in consumers of fresh leafy greens. Viability assays are essential to accurately estimate health risk caused by viable parasites that contaminate food. We evaluated the efficacy of reverse transcription quantitative PCR (RT-qPCR), propidium monoazide coupled with (q)PCR, and viability staining using propidium iodide through systematic laboratory spiking experiments for selective detection of viable Cryptosporidium parvum, Giardia enterica, and Toxoplasma gondii. In the presence of only viable protozoa, the RT-qPCR assays could accurately detect two to nine (oo)cysts/g spinach (in 10 g processed). When different proportions of viable and inactivated parasite were spiked, mRNA concentrations correlated with increasing proportions of viable (oo)cysts, although low levels of false-positive mRNA signals were detectable in the presence of high amounts of inactivated protozoa. Our study demonstrated that among the methods tested, RT-qPCR performed more effectively to discriminate viable from inactivated C. parvum, G. enterica and T. gondii on spinach. This application of viability methods on leafy greens can be adopted by the produce industry and regulatory agencies charged with protection of human public health to screen leafy greens for the presence of viable protozoan pathogen contamination.

Azathioprine-induced toxoplasma gondii infection in a patient with Crohn's disease with NUDT15 variation: A case report.

INTRODUCTION: Azathioprine (AZA) has been widely used for the treatment of various immune-related diseases and has become a mainstay in the treatment of inflammatory bowel disease. However, patients with genetic mutations may experience severe adverse events when treated with azathioprine. Most of the previous literature focused on the TPMP gene-related adverse reactions, herein, we report a case of Crohn's disease patient with nucleoside diphosphate-linked moiety X motif 15 gene (NUDT15) variation and wild-type TPMP gene who developed toxoplasma gondii infection after azathioprine treatment. PATIENT CONCERNS: A 56-year-old Crohn's disease patient developed toxoplasma gondii infection within 2 months after the administration of azathioprine; however, he had no relevant high-risk factors. DIAGNOSIS: Subsequent genetic testing revealed that the patient was heterozygous for NUDT15. Therefore, it was reasonable to consider that the patient's genetic mutation resulted in reduced tolerance to azathioprine, leading to low immunity and eventually toxoplasma infection. INTERVENTIONS: AZA was then discontinued; after anti-infection, antipyretic and other supportive treatments were administered, the patient's condition gradually improved. OUTCOMES: The patient was followed up at 1, 3, and 6 months after discharge; fortunately, he was in good health. CONCLUSION: We report a case of Crohn's disease in a patient who developed severe pneumonia caused by toxoplasma gondii infection due to the administration of AZA, with normal TPMP gene but NUDT15 gene mutation. This indicates that NUDT15 variation may contribute to severe adverse events in patients treated with azathioprine, and we suggest that NUDT15 genotype be detected before the use of azathioprine in order to provide personalized therapy and reduce side effects.

Endoparasite prevalence and infection risk factors among cats in an animal shelter in Estonia.

Cats are important hosts for different zoonotic parasites that can be hazardous to human health. To date, few studies have attempted to identify the factors affecting parasitic infections in shelter animals. This study aims to analyse the presence of endoparasites in shelter cats in Tartu, Estonia, and identify factors affecting endoparasite prevalence and intensity. The risk factors considered were age, location (urban vs rural cats) and time spent in shelter. In total, 290 faecal samples were collected from cats at an animal shelter in 2015-2016 and investigated for endoparasites using the concentration flotation technique. In total, 138 shelter cats (47.6%) were infected with endoparasites and their overall prevalence was: Toxocara cati (36.6%), Cystoisospora spp. (12.4%), Taeniidae gen. sp. (4.1%), Toxoplasma gondii/Hammondia hammondi (3.4%), Eucoleus aerophilus (2.1%), Cryptosporidium spp. (2.1%), Ancylostoma sp. (0.7%) and Giardia sp. (0.7%). Coinfections occurred in 38 cats (13.1%) most frequently of T. cati and Cystoisospora spp. (4.5%), Cystoisospora spp. and T. gondii/H. hammondi (2.1%). Where species identification of cestode and nematode samples was not possible according to morphology, genetic analysis of the mitochondrial cox1 gene was carried out. DNA was successfully analysed for 6 out of 13 samples that required genetic identification, revealing Ancylostoma tubaeforme in one nematode sample and Hydatigera taeniaeformis in five cestode samples. Cats from rural areas had significantly higher endoparasite prevalence than cats from urban areas. Helminth prevalence decreased to some extent due to anthelmintic treatment in cats available for adoption (held ≥15 days in the shelter), whereas the prevalence of infection with protists increased significantly in these animals. It is important to note that the analysis revealed lower infection intensity for quarantine cats (held 1-14 days in the shelter) compared with cats available for adoption. The relatively high prevalence of endoparasites (including zoonotic) in shelter cats ready for adoption suggests that current anthelminthic procedures require improvements.

Outbreaks of clinical toxoplasmosis in humans: five decades of personal experience, perspectives and lessons learned.

BACKGROUND: The protozoan parasite Toxoplasma gondii has a worldwide distribution and a very wide host range, infecting most warm-blooded hosts. Approximately 30% of humanity is infected with T. gondii, but clinical toxoplasmosis is relatively infrequent. Toxoplasmosis has a wide range of clinical symptoms involving almost all organ systems. In most persons that acquire infection postnatally, symptoms (when present) are mild and mimic other diseases such as flu, Lyme disease, Q fever, hematological alterations, or mumps. It is likely that clinical disease is more common than reported. The ingestion of infected meat or food and water contaminated with oocysts are the two main modes of postnatal transmission of Toxoplasma gondii. The infective dose and the incubation period of T. gondii infection are unknown because there are no human volunteer experiments. METHODS: Here, I have critically reviewed outbreaks of clinical toxoplasmosis in humans for the past 55 years, 1966-2020. Information from oocyst-acquired versus meat-acquired infections was assessed separately. RESULTS: Most outbreaks were from Brazil. There were no apparent differences in types or severity of symptoms in meat- versus oocyst-acquired infections. Fever, cervical lymphadenopathy, myalgia, and fatigue were the most important symptoms, and these symptoms were not age-dependent. The incubation period was 7-30 days. A genetic predisposition to cause eye disease is suspected in the parasites responsible for three outbreaks (in Brazil, Canada, and India). Only a few T. gondii tissue cysts might suffice to cause infection, as indicated by outbreaks affecting some (but not all) individuals sharing a meal of infected meat. CONCLUSIONS: Whether the high frequency of outbreaks of toxoplasmosis in humans in Brazil is related to environmental contamination, poor hygiene, socioeconomic conditions, or to genotypes of T. gondii needs investigation.

Comparison between Enzyme Immunoassays Performed on Samples of Dried Blood and Serum for Toxoplasmosis Prenatal Screening: Population-based Study / Comparação entre ensaios imunoenzimáticos realizados em amostras de sangue seco e soro para triagem prénatal da toxoplasmose: Estudo populacional

Abstract Objective Most prenatal screening programs for toxoplasmosis use immunoassays in serum samples of pregnant women. Few studies assess the accuracy of screening tests in dried blood spots, which are of easy collection, storage, and transportation. The goals of the present study are to determine the performance and evaluate the agreement between an immunoassay of dried blood spots and a reference test in the serum of pregnant women from a population-based prenatal screening program for toxoplasmosis in Brazil. Methods A cross-sectional study was performed to compare the immunoassays Imunoscreen Toxoplasmose IgM and Imunoscreen Toxoplasmose IgG (Mbiolog Diagnósticos, Ltda., Contagem, Minas Gerais, Brazil)in dried blood spots with the enzymelinked fluorescent assay (ELFA, BioMérieux S.A., Lyon, France) reference standard in the serum of pregnant women from Minas Gerais Congenital Toxoplasmosis Control Program. Results The dried blood spot test was able to discriminate positive and negative results of pregnant women when comparedwith the reference test, with an accuracy of 98.2% for immunoglobulin G (IgG), and of 95.8% for immunoglobulin M (IgM). Conclusion Dried blood samples are easy to collect, store, and transport, and they have a good performance,making this a promisingmethod for prenatal toxoplasmosis screening programs in countries with continental dimensions, limited resources, and a high prevalence of toxoplasmosis, as is the case of Brazil.

Resumo Objetivo A maioria dos programas de triagem pré-natal para toxoplasmose utiliza imunoensaios em amostras de soro de gestantes. Poucos estudos avaliam a acurácia dos testes de triagem em amostras de sangue seco, que são de fácil coleta, armazenamento e transporte. Este estudo teve como objetivo determinar o desempenho e avaliar a concordância entre um imunoensaio em sangue seco e um teste de referência em soro de gestantes de um programa de rastreamento pré-natal de base populacional para toxoplasmose no Brasil. Métodos Realizou-se um estudo transversal para comparar os imunoensaios Imunoscreen Toxoplasmose IgM e Imunoscreen Toxoplasmose IgG (Mbiolog Diagnósticos, Ltda., Contagem, Minas Gerais, Brazil) em sangue seco com o padrão de referência ensaio fluorescente ligado a enzimas (enzyme-linked fluorescent assay, ELFA, BioMérieux S.A., Lion, França) no soro de gestantes do Programa de Controle de Toxoplasmose Congênita de Minas Gerais. Resultados O exame em sangue seco foi capaz de discriminar os resultados positivos e negativos das gestantes quando comparado ao teste de referência, com acurácia de 98,2% para imunoglobulina G (IgG), e de 95,8% para imunoglobulina M (IgM). Conclusão O sangue seco apresenta bom desempenho e é uma amostra de fácil coleta, armazenamento e transporte, o que o torna um método promissor para programas de triagem pré-natal de toxoplasmose em países com dimensões continentais, recursos limitados, e alta prevalência de toxoplasmose, como é o caso do Brasil.

Epidemiologic and Public Health Significance of Toxoplasma gondii Infections in Venison: 2009-2020.

Toxoplasma gondii infections are common in humans and animals worldwide. The ingestion of food or water contaminated with oocysts excreted by infected cats or ingestion of uncooked or undercooked meat containing tissue cysts of T. gondii are the 2 major modes of transmission of T. gondii. Deer are a popular game. Recently, outbreaks of clinical toxoplasmosis were reported in humans in North America linked to ingestion of undercooked venison. Here, we review prevalence, persistence of infection, clinical disease, epidemiology, and public health risks of T. gondii infections in deer and other cervids for the past decade. Estimates of worldwide serological prevalence are summarized individually for each species of deer, elk, moose, and caribou. Genetic diversity of 112 viable isolates of T. gondii from cervids is discussed, including its public health significance. Prevalence of T. gondii in deer is very high. Any part of a deer, including liver, spleen, and muscles, should be cooked thoroughly before human consumption.

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.

Study on prevalence and liver function test enzymes of differently plumaged peafowls (Pavo cristatus) infected with Toxoplasma gondii in captivity / [Estudo sobre enzimas de prevalência e teste da função hepatica de pavões de plumage diferente (Pavo cristatus) infectados com Toxoplasma gondii em cativeiro]

O presente estudo foi realizado para determinar a prevalência geral de toxoplasmose em pavões de plumagem diferente e seu efeito nas enzimas de teste da função hepática dos hospedeiros. Um total de cem pavões de plumas diferenciais, como ombro preto (n = 52), azul (n = 28), branco (n = 10) e arlequim (n = 10) foram estudados no zoológico de Bahawalpur, no Paquistão, usando o Latex Agglutination Test (LAT) e ensaio imunossorvente ligado a enzima (ELISA). A prevalência geral por LAT e ELISA foi de 37% e 30%, respectivamente. Por LAT, observou-se uma prevalência não significativamente maior (P≥0,05) em gênero (37,77%) nos machos do que nas fêmeas (36,36%), enquanto os adultos apresentaram uma prevalência maior (37,97%) em relação aos jovens (33,33%). De acordo com o ELISA, uma prevalência significativamente (P <0,05) maior (35,55%) foi observada nos machos do que nas fêmeas (25,45%) e significativamente (P <0,05) maior prevalência (31,64%) foi registrada nos adultos do que nos jovens (23,80%). A análise do perfil bioquímico sérico mostrou que o nível de bilirrubina não teve elevação significativa nos hospedeiros infectados, em comparação aos não infectados, enquanto a concentração de albumina, alanina aminotransferase (ALT), aspartato aminotransferase (AST), fosfatase alcalina (ALP) foi significativamente (P <0,05) diferente nos hospedeiros infectados. Conclui-se que a toxoplasmose afeta as enzimas do teste da função hepática. Essa é uma pesquisa preliminar e requer mais pesquisas em todo o país, com populações e amostras maiores.(AU)

A streamlined clinical metagenomic sequencing protocol for rapid pathogen identification.

Metagenomic next-generation sequencing (mNGS) holds promise as a diagnostic tool for unbiased pathogen identification and precision medicine. However, its medical utility depends largely on assay simplicity and reproducibility. In the current study, we aimed to develop a streamlined Illumina and Oxford Nanopore-based DNA/RNA library preparation protocol and rapid data analysis pipeline. The Illumina sequencing-based mNGS method was first developed and evaluated using a set of samples with known aetiology. Its sensitivity for RNA viruses (influenza A, H1N1) was < 6.4 × 102 EID50/mL, and a good correlation between viral loads and mapped reads was observed. Then, the rapid turnaround time of Nanopore sequencing was tested by sequencing influenza A virus and adenoviruses. Furthermore, 11 respiratory swabs or sputum samples pre-tested for a panel of pathogens were analysed, and the pathogens identified by Illumina sequencing showed 81.8% concordance with qPCR results. Additional sequencing of cerebrospinal fluid (CSF) samples from HIV-1-positive patients with meningitis/encephalitis detected HIV-1 RNA and Toxoplasma gondii sequences. In conclusion, we have developed a simplified protocol that realizes efficient metagenomic sequencing of a variety of clinical samples and pathogen identification in a clinically meaningful time frame.

Disseminated Toxoplasmosis in a Narrow-Ridged Finless Porpoise (Neophocaena asiaeorientalis) with Transplacental Embryonal Transmission.

We describe a case of systemic toxoplasmosis in a female adult narrow-ridged finless porpoise (Neophocaena asiaeorientalis) found in May 2018 inside a gillnet set in the Ariake Sound, southern Japan. The main lesions observed were lymphoplasmacytic and focally necrotizing encephalitis, necrotizing to granulomatous adrenalitis, myocarditis, and inflammation in the intestinal wall, associated with protozoal tissue cysts and tachyzoites. Additionally, the individual had a 5.6 mm (crown-rump length) early-stage embryo in the left uterine horn, which had multifocal necrotizing lesions with intralesional tissue cysts and tachyzoites in the parenchyma. Immunohistochemistry and PCR and sequencing of the internal transcribed spacer 1 region confirmed a Toxoplasma gondii infection. Further genotyping revealed an atypical type II genotype with a type I pattern for the Apico locus. Narrow-ridged finless porpoises are an endangered coastal species already facing various anthropogenic threats. Toxoplasmosis, especially with its ability to transmit to an early-stage embryo, should be considered an emerging threat to this vulnerable species.

Serological Determination of Toxoplasma gondii among Sheep (Ovis aries) in Guilan Province, Iran.

Toxoplasma gondii is one of the most common foodborne protozoan parasite causing congenital infection, abortion, and stillbirth in humans and animals. The temperate and humid climate is one of the most important factors in the high prevalence of T. gondii. Sheep are among the important sources of meat production in Guilan province, Iran. Therefore, the consumption of raw and half-cooked meat is one of the major risk factors for T. gondii infection. Toxoplasmosis in patients with intact immune systems is usually asymptomatic; however, it but can be life-threatening in patients with a weak immune system (for example, patients with the human immunodeficiency viruses/acquired immunodeficiency syndrome or cancer and transplant recipients). Guilan is divided into three geographical regions of plains with a temperate climatic condition, hillsides with a semi-humid climate, and heights with cold mountainous climate. Climate situations play a role in the prevalence of toxoplasmosis. The present study aimed to investigate the seroprevalence of T. gondii infection among sheep in Guilan province, north of Iran. In the current cross-sectional study, a total of 400 sheep sera samples were tested for the determination of immunoglobulin G (IgG) antibody against T. gondii using the enzyme-linked immunosorbent assay. The samples were divided into different groups according to the geographical location and animal age. T. gondii antibody (i.e., IgG) was detected in 166 sheep (41.5%). The highest frequency of T. gondii infection (72.7%; n=56) was observed for the age group of &gt; 4 years; the difference was statistically significant in this regard (P=0.0001) in comparison to that reported for other groups. In addition, the seroprevalence of T. gondii was significantly higher in the plains (53.9%) than that of the hillsides and heights (P=0.0001). Consequently, the seroprevalence of T. gondii infection in Guilan was high indicating a significant relationship with geographical location and animal age.

Diffuse Unilateral Subacute Neuroretinitis Evolving With Submacular Granuloma.

DUSN is an infectious ocular disease that can lead to severe visual impairment and blindness. It usually occurs in young healthy individuals and depending on the stage of the disease, clinical presentation may range from mild vitritis and multifocal gray-white lesions in outer retina to optic atrophy.Parasites of different sizes and species have been proposed as the etiological agent of this disease. Thus, it is hypothesized that different infectious worms may be considered as the likely cause of a both autoimmune and toxic form of nematode retinopathy.Most patients present with already severe visual impairment and in the later stages of the disease, where the likelihood of improvement is low, despite therapy. In cases of early diagnosis, prompt treatment, whether with oral antihelmintic or direct photocoagulation of the worm, patients may show considerable visual improvement and have a more favorable prognosis.

Evaluation of Apoptosis, Proliferation, and Adhesion Molecule Expression in Trophoblastic Tissue of Women With Recurrent Spontaneous Abortion and Infected With Toxoplasma gondii.

Recurrent spontaneous abortion is an obstetric complication with undefined causes. Apoptosis, proliferation, and adhesion are considered important factors in the pathogenesis of abortion. This work aimed to determine Bax and Bcl-2 as a proapoptotic and antiapoptotic protein, Ki67 and P27kip as proliferative and antiproliferative proteins, and E-cadherin and CD44 as adhesion molecules in the trophoblastic tissues in cases with recurrent miscarriage. Immunohistochemistry and quantitative polymerase chain reaction analysis of Bax, Bcl-2, Ki67, P27kip, E-cadherin, and CD44 in paraffin-embedded sections of placental tissues obtained from 108 women were divided into 3 categories: 66 Toxoplasma gondii-positive women with recurrent abortion, 22 T. gondii-negative women with recurrent abortion, and 20 women with no history of abortion as a control group. The mean ratio of the expression of Bax and P27kip proteins was 35.3% and 36.1%, which is significantly higher than that of the second group (19.88 and 20.02%), and the third group (12.3% and 10.98%), while the mean ratio of the expression of Bcl-2, Ki67, E-cadherin, and CD44 proteins was 12.35%, 11.23%, 10.32%, and 9.97%, which is significantly lower than that of the second group (33.75%, 13.18%, 21.88%, and 23.29%) and that of the third group (38.58%, 39.27%, 37.98%, and 35.79%). The presence of proapoptotic protein (Bax) and antiproliferative protein (P27kip) at high levels and the presence of antiapoptotic protein (Bcl-2), proliferative protein (Ki67), and adhesion molecules (E-cadherin and CD44) in lower levels in the T. gondii-positive group clarify the mechanism involved in the induction of abortion and loss of pregnancy.

Characterization of a spontaneous cyst-forming strain of Toxoplasma gondii isolated from Tokachi subprefecture in Japan.

Apicomplexan parasite Toxoplasma gondii has three distinct clonal lineages: high, medium and low virulent strains, type I, II and III, respectively. T. gondii avoids the immune response by transforming from fast multiplying tachyzoite to slow multiplying bradyzoite, and establishing a chronic infection. In the present study, we isolated a new strain of T. gondii from cat feces in the Tokachi subprefecture, Hokkaido, Japan and named it as TgCatJpObi1 (Obi1) strain. Genotyping analysis of 12 loci revealed atypical characters close to type II, genotype 4 according to ToxoDB classification. Phenotypically, Obi1 strain shows slow growth rate and the ability of spontaneous cyst formation in both human foreskin fibroblast (HFFs) and mouse peritoneal macrophages in vitro without bradyzoite induction. Parasite virulence was assessed by means of mouse survival upon infection with either Obi1 or ME49 strains. Obi1 strain displayed no mortalities in comparison to type II clonal lineage, ME49 at LD50 to LD100 range (1 × 103-106 tachyzoites). Although virulence of Obi1 strain is significantly lower than that of ME49, nucleotide sequences analyses revealed that genes of virulence factors such as Gra15, Rop5, 16, 17, and 18 in Obi1 strain were 100% identical to those in the type II strain. Thus, characterization of a newly isolated strain, Obi1, is crucial to clarify the development of toxoplasmosis in both humans and animals.