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.

The involvement of protozoan parasites in sheep abortions - A ten-year review of diagnostic results.

Abortions in sheep flocks affect animal health and lead to significant losses in productivity, with severe economic consequences. In recent years, the role of protozoan parasites as the cause of ovine abortions has been significant. Here, the diagnosis of infectious causes of abortions in sheep in Israel in the last decade is reviewed, focusing on parasitic pathogens. Analysis of the serological data of sheep sera (including aborted fetuses) submitted for diagnoses between 2010 and 2019 revealed overall seroprevalence of 67.4 % and 46.7 % for Neospora spp. and Toxoplasma gondii respectively, with high rates of co-exposure (32.4 %). The seroprevalence of T. gondii was higher in aborting ewes than in pre-sale examinations (48.2 % and 28.9 %, respectively (P < 0.001)). The seroprevalence of Neospora spp. was significantly higher than the seroprevalence of Toxoplasma (P < 0.001), and was similar in samples from aborting ewes and in samples from pre-sale examinations. In addition, the presence of anti-Neospora spp. antibodies was the most prominent finding diagnosed in aborted fetuses (22.9 % of aborted fetuses, significantly higher than any other organism, P < 0.001). The results of this study demonstrate that in endemic areas the seroprevalence of N. caninum in sheep may be high, and should be considered as an important cause of abortions. However, since the seroprevalence is high even in non-aborting ewes, in order to determine the causative agent of abortion in endemic flocks, a comprehensive epidemiological investigation is warranted.

Retrospective study of toxoplasmosis prevalence in pregnant women in Benin and its relation with malaria.

BACKGROUND: Globally distributed with variable prevalence depending on geography, toxoplasmosis is a zoonosis caused by an obligate intracellular protozoan parasite, Toxoplasma gondii. This disease is usually benign but poses a risk for immunocompromised people and for newborns of mothers with a primary infection during pregnancy because of the risk of congenital toxoplasmosis (CT). CT can cause severe damage to fetuses-newborns. To our knowledge, no study has been conducted in sub-Saharan Africa on toxoplasmosis seroprevalence, seroconversion and CT in a large longitudinal cohort and furthermore, no observation has been made of potential relationships with malaria. METHODS: We performed a retrospective toxoplasmosis serological study using available samples from a large cohort of 1,037 pregnant women who were enrolled in a malaria follow-up during the 2008-2010 period in a rural area in Benin. We also used some existing data to investigate potential relationships between the maternal toxoplasmosis serological status and recorded malaria infections. RESULTS: Toxoplasmosis seroprevalence, seroconversion and CT rates were 52.6%, 3.4% and 0.2%, respectively, reflecting the population situation of toxoplasmosis, without targeted medical intervention. The education level influences the toxoplasmosis serological status of women, with women with little or no formal education have greater immunity than others. Surprisingly, toxoplasmosis seropositive pregnant women tended to present lower malaria infection during pregnancy (number) or at delivery (presence) and to have lower IgG levels to Plasmodium falciparum Apical Membrane Antigen 1, compared to toxoplasmosis seronegative women. CONCLUSIONS: The high toxoplasmosis seroprevalence indicates that prevention against this parasite remains important to deploy and must be accessible and understandable to and for all individuals (educated and non-educated). A potential protective role against malaria conferred by a preexisting toxoplasmosis infection needs to be explored more precisely to examine the environmental, parasitic and/or immune aspects.

Posterior segment findings by spectral-domain optical coherence tomography and clinical associations in active toxoplasmic retinochoroiditis.

Toxoplasmic retinochoroiditis is a common, potentially blinding parasitic infection. We sought to define the spectrum and frequency of signs of active toxoplasmic retinochoroiditis by spectral domain optical coherence tomography (SD-OCT), and to identify clinical associations. Ninety eyes of 90 individuals presenting consecutively to a tertiary referral uveitis service with active toxoplasmic retinochoroiditis and gradable SD-OCT scans were evaluated prospectively. SD-OCT features were collated, and associations with lesion location, primary versus recurrent episode, serological status, human immunodeficiency virus infection and best-corrected Snellen visual acuity were explored. Active toxoplasmic retinochoroiditis presented with thickened (65%) and hyperreflective (61%) retina, choroidal thickening (55%) and hyporeflectivity (61%), hyperreflective vitreous dots (80%) and deposits (36%), and posterior hyaloid thickening (35%) on SD-OCT. Most signs occurred with similar frequency across clinical groups. Retinal hyporeflectivity (17%) was significantly associated with a visual acuity of 20/200 or worse at resolution. Our observations demonstrate that active toxoplasmic retinochoroiditis has diverse SD-OCT signs and that none are universally present. Retinal hyporeflectivity-suggesting liquefactive necrosis-predicts poor visual outcome.

Molecular detection of Toxoplasma gondii in placentas of women who received therapy during gestation in a toxoplasmosis outbreak.

Toxoplasmosis is a disease caused by T. gondii, a protozoa which affects humans and animals and is widely distributed worldwide. In humans, there is great concern due to the serious consequences that can occur in the infection of pregnant women and the newborn. The early diagnosis of gestational toxoplasmosis is important for treatment to be carried out in order to prevent vertical transmission or reduce damage. The diagnosis can be made through the detection of antibodies in pregnant women or neonates and PCR of amniotic fluid. Previous studies have also reported PCR of the placenta as a good diagnostic test. Our study evaluated the detection of T. gondii DNA in placenta samples from parturients seen at the University Hospital of Santa Maria, Southern Brazil and treated during the pregnancy. We performed PCR in forty samples and five were positive, representing 12.5%. When correlating the treatment time and the detection of DNA in the placentas, no significant result was found. The prevalence of positive samples was lower than in other studies in the literature. The data reaffirm the importance of carrying out the analysis of the placenta.

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.

Detection of Pneumocystis jirovecii and Toxoplasma gondii in patients with lung infections by a duplex qPCR assay.

Pneumocystis pneumonia (PCP) and pulmonary toxoplasmosis (PT) are caused by Pneumocystis jirovecii and Toxoplasma gondii. The clinical symptoms and imaging of PCP and PT are indistinguishable. A duplex qPCR was developed to differentiate between these two pathogens. In testing 92 clinical samples to validate the performance of this method for P. jirovecii detection, it identified 31 positive samples for P. jirovecii infection, consistent with clinical diagnosis. Among the remainder of the 61 clinical samples with suspected PCP, yet showing as negative by the conventional PCR diagnosis approach, 6 of them proved positive using our new assay. Our new approach also produced similar results in identification of T. gondii infections, giving a result of 2 positive and 20 negative in clinical samples. An investigation was undertaken on the prevalence of P. jirovecii and T. gondii infections using 113 samples from lung infection patients. 9% (10/113) were shown to be positive with infections of P. jirovecii, 2% with T. gondii (2/113) and 5% (6/113) were co-infected with both pathogens. Although this duplex qPCR can detect individual P. jirovecii and T. gondii infection, and co-infection of both pathogens, further large-scale investigations are needed to validate its performance, especially in T. gondii detection. Our assay provides a rapid and accurate tool for PCP and PT diagnosis in immunocompromised population and clinical surveillance of these infections in patients with no immune defects.

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.

Detection of toxoplasmosis in pets and stray cats through molecular and serological techniques in Khyber Pakhtunkhwa, Pakistan.

BACKGROUND: Toxoplasma gondii is an important parasite that belongs to the phylum Apicomplexa, distributed globally, causing major health issues for a wide range of hosts, including humans, native and wild animals. METHODS: In the present study, we detected IgG and IgM antibodies through an ELISA kit and DNA of T. gondii through PCR in 197 pets and stray cats in Peshawar, Charsadda, Mardan, and Kohat districts of Khyber Pakhtunkhwa (Pakistan) to estimate the existence of feline toxoplasmosis. RESULTS: The current study revealed that stray cats have a significant infection rate of T. gondii (74.6%) as compared to pet cats (25.4%). In all the four districts, the prevalence of T. gondii was pointedly higher in district Kohat (95.5%) in the feline population. In comparison to the female (75.18%) and male (both pets and stray) cats have a maximum infection of (81.66%) non-significantly. The prevalence of T. gondii was observed to be significantly higher (91.66%) in the older and greater than 4 year old population of cats as compared to the younger ones. In poor health condition, the cat populations has a higher risk of infection of 92.3% as compared to healthy and poor body condition (73.91%) and (82.6%) respectively. The chronic and reactivated chronic conditions of toxoplasmosis were higher (58.37%) as compared to the acute condition. CONCLUSION: It has been concluded that toxoplasmosis is widely spread in the studied population.The outcomes of the present study show that T. gondii infection has a significant impact on the type of cat, age, and area, which implies a serious threat to human beings. Therefore, genotyping of T. gondii strains from different hosts is needed to forecast the current approach for prevention and control of this zoonotic parasite.

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.

Toxoplasma gondii Tyrosine-Rich Oocyst Wall Protein: A Closer Look through an In Silico Prism.

Toxoplasmosis is a global threat with significant zoonotic concern. The present in silico study was aimed at determination of bioinformatics features and immunogenic epitopes of a tyrosine-rich oocyst wall protein (TrOWP) of Toxoplasma gondii. After retrieving the amino acid sequence from UniProt database, several parameters were predicted including antigenicity, allergenicity, solubility and physico-chemical features, signal peptide, transmembrane domain, and posttranslational modifications. Following secondary and tertiary structure prediction, the 3D model was refined, and immunogenic epitopes were forecasted. It was a 25.57 kDa hydrophilic molecule with 236 residues, a signal peptide, and significant antigenicity scores. Moreover, several linear and conformational B-cell epitopes were present. Also, potential mouse and human cytotoxic T-lymphocyte (CTL) and helper T-lymphocyte (HTL) epitopes were predicted in the sequence. The findings of the present in silico study are promising as they render beneficial characteristics of TrOWP to be included in future vaccination experiments.

Cross-sectional association of Toxoplasma gondii exposure with BMI and diet in US adults.

Toxoplasmosis gondii exposure has been linked to increased impulsivity and risky behaviors, which has implications for eating behavior. Impulsivity and risk tolerance is known to be related with worse diets and a higher chance of obesity. There is little known, however, about the independent link between Toxoplasma gondii (T. gondii) exposure and diet-related outcomes. Using linear and quantile regression, we estimated the relationship between T. gondii exposure and BMI, total energy intake (kcal), and diet quality as measured by the Health Eating Index-2015 (HEI) among 9,853 adults from the 2009-2014 National Health and Nutrition Examination Survey. Previous studies have shown different behavioral responses to T. gondii infection among males and females, and socioeconomic factors are also likely to be important as both T. gondii and poor diet are more prevalent among U.S. populations in poverty. We therefore measured the associations between T. gondii and diet-related outcomes separately for men and women and for respondents in poverty. Among females <200% of the federal poverty level Toxoplasmosis gondii exposure was associated with a higher BMI by 2.0 units (95% CI [0.22, 3.83]) at median BMI and a lower HEI by 5.05 units (95% CI [-7.87, -2.24]) at the 25th percentile of HEI. Stronger associations were found at higher levels of BMI and worse diet quality among females. No associations were found among males. Through a detailed investigation of mechanisms, we were able to rule out T. gondii exposure from cat ownership, differing amounts of meat, and drinking water source as potential confounding factors; environmental exposure to T. gondii as well as changes in human behavior due to parasitic infection remain primary mechanisms.

High prevalence of anti-Toxoplasma gondii antibodies in beef cattle in Midwestern Brazil.

The present study aimed to determine the seroprevalence of anti-Toxoplasma gondii antibodies using an indirect immunofluorescence assay (IFA-IgG) in 3,814 cows aged ≥ 24 months belonging to 353 beef farms in the Mato Grosso state. Additionally, we aimed to identify the possible risk factors associated with seropositivity in Mato Grosso, which encompasses three biomes (Amazon, Cerrado, and Pantanal) of Brazil. Analysis of 3,814 samples observed that 1,307 animals were positive for anti-T. gondii antibodies (IFA-IgG ≥ 64), with an animal-level seroprevalence of 34.27%. Herd-level seroprevalence was 92.07%. In the animal-level model, cows raised in the Amazon and Pantanal biomes and breed studies on European and Zebu or hybrid were found to be at risk for T. gondii seropositivity. Prevalence of anti-T. gondii antibodies in cows destined for human consumption was found to be widely distributed throughout the entire study area. Further studies are required to assess the impact of beef in the possible transmission of toxoplasmosis to humans residing in the region and establish additional control measures for this protozoan mainly on beef herds raised in the Amazon and Pantanal biomes, where the highest values of seroprevalence were observed.

Prevalence and risk factors of Toxoplasma gondii and Leishmania spp. infections in apparently healthy dogs in west Shewa zone, Oromia, Ethiopia.

BACKGROUND: In urban settings, the presence of a high density of the human population and contact with domestic and/or stray animals such as dogs and cats can be risk factors for the transmission of zoonotic protozoa parasites. Toxoplasma gondii (T. gondii) and Leishmania spp. are zoonotic protozoon parasites with significant health burdens worldwide. METHODS: A cross-sectional study was used to investigate the antibody prevalence and risk factors of T. gondii and Leishmania spp. infections in 385 randomly selected dogs of Ambo, Bako, and Gojo towns of West Shewa Zone, Oromia regional state, Ethiopia. A questionnaire survey was administered to households to collect data on potential risk factors. Dog sera samples were assayed for T. gondii IgG antibodies using the direct agglutination test while Leishmania spp. specific antibodies tested using an indirect enzyme-linked immunosorbent assay (ELISA). Logistic regression was used for data analysis. RESULTS: Overall, T. gondii and Leishmania spp. infection seroprevalence was found to be 82.86% (95% confidence interval (CI): 78.71-86.49%) and 92.47% (95% CI: 89.36-94.90%), respectively. Seropositivity for both T. gondii and Leishmania spp. was found in 82.58% of the dogs. None of the investigated factors were associated with Leishmania spp. seropositivity (p > 0.05). The seroprevalence of T. gondii was significantly different among the study towns (p = 0.003). The risk of T. gondii infection was 2.71 times higher in adult dogs than juvenile dogs (p = 0.043). Dogs kept simultaneously with other domestic animals had increased odds of T. gondii seropositivity compared to those with no other domestic animals (Adjusted Odds ratio: 1.96, p = 0.021). However, altitude, sex, breed, housing, feeding, educational level of head of the household, and dog's living area were not significantly associated with T. gondii seropositivity (p > 0.05). CONCLUSIONS: The high seropositivity and the simultaneous presence of antibodies of T. gondii and Leishmania spp. in dogs suggest the widespread nature of these parasites in the environment and the high potential of transmission to other animals and humans. Further epidemiological studies, isolation and molecular characterization of the parasites, and educational campaigns are suggested.

Herd-level contamination of Neospora caninum, Toxoplasma gondii and Brucella in milk of Iranian dairy farms.

The bulk milk examination is a reliable screening tool for monitoring the quality of milk in the farms. The infection to Neospora caninum, Toxoplasma gondii and Brucella sp. Was evaluated in bulk milk samples of dairy farms in Hamedan province, West part of Iran. All the dairy farms (n = 149) were examined for N. caninum, T. gondii and Brucella infections using milk ring test (MRT), microbiology, serology (Enzyme-linked Immunosorbent Assay), and molecular techniques. Based on molecular methods, Brucella-infection was negative in all farms; while, 55 %, 5.4 % and 2.7 % of samples were positive for N. caninum, T. gondii and mix infection, respectively. The highest Neospora-infection was detected in the farms with history of abortion in fall and winter. There was significant association between Neospora-infection and the presence of dogs and rodents in the farms, herd size, and age of the animals. Also, a significant association was seen between Toxoplasma-infection and the presence of cats and rodents in the farms, as well as age of the animals. Average total bacterial count (TBC) was calculated 1.14 × 106±1.1 × 106. The highest TBC was in the farms from Central locations of studied area (5.7 × 106±2.24 × 106), farms with more than 120 animals (7.9 × 106±2.8 × 106), and farms with ≥50-months age (1.74 × 106±6.3 × 105) in spring and summer (6.9 × 106±3.7 × 106). The number of somatic cells was estimated between 1 × 104 and 2 × 106 (Average = 4.2 × 105±3.39 × 105). The current study was a comprehensive evaluation of Neospora, Toxoplasma and Brucella infections in milk samples of Iranian dairy farms for the first time. Neospora-infection is responsible for economic losses in the region. Health education and milk pasteurization are so helpful for inhibiting the milk borne diseases. To reduce the risk factors, predict and design the appropriate schemes like redundant of heterogeneous animals are recommended.

Effector Vγ9Vδ2 T cell response to congenital Toxoplasma gondii infection.

A major γδ T cell population in human adult blood are the Vγ9Vδ2 T cells that are activated and expanded in a TCR-dependent manner by microbe-derived and endogenously derived phosphorylated prenyl metabolites (phosphoantigens). Vγ9Vδ2 T cells are also abundant in human fetal peripheral blood, but compared with their adult counterparts they have a distinct developmental origin, are hyporesponsive toward in vitro phosphoantigen exposure, and do not possess a cytotoxic effector phenotype. In order to obtain insight into the role of Vγ9Vδ2 T cells in the human fetus, we investigated their response to in utero infection with the phosphoantigen-producing parasite Toxoplasma gondii (T. gondii). Vγ9Vδ2 T cells expanded strongly when faced with congenital T. gondii infection, which was associated with differentiation toward potent cytotoxic effector cells. The Vγ9Vδ2 T cell expansion in utero resulted in a fetal footprint with public germline-encoded clonotypes in the Vγ9Vδ2 TCR repertoire 2 months after birth. Overall, our data indicate that the human fetus, from early gestation onward, possesses public Vγ9Vδ2 T cells that acquire effector functions following parasite infections.

Determination of parasitic burden in the brain tissue of infected mice in acute toxoplasmosis after treatment by fluconazole combined with sulfadiazine and pyrimethamine.

BACKGROUND/AIMS: One of the opportunistic pathogens which cause serious problems in the human immune system is Toxoplasma gondii, with toxoplasma encephalitis (TE) seen in patients affected by it. The treatment of these patients is limited, and if not treated on time, death will be possible. METHODS: In this study, the effects of the treatment with different doses of fluconazole (FLZ) in combination with the current treatment of acute toxoplasmosis on reducing the mortality rate and the parasitic load in the murine model in vivo were studied. The mice were treated with different doses of fluconazole alone, sulfadiazine, and pyrimethamine plus fluconazole. A day after the end of the treatment and 1 day before death, the mice's brains were collected, and after DNA extraction and molecular tests, the parasite burden was detected. RESULTS: This study showed that a 10-day treatment with 20 mg/kg of fluconazole combined with sulfadiazine and pyrimethamine 1.40 mg/kg per day affected acute toxoplasmosis and reduced the parasitic load significantly in brain tissues and also increased the survival rate of all mice in this group until the last day of the study, in contrast to other treatment groups. These results also indicate the positive effects of combined therapy on Toxoplasma gondii and the prevention of relapse. CONCLUSIONS: Reducing the parasitic burden and increasing the survival rate were more effective against acute toxoplasmosis in the combined treatment of different doses of fluconazole with current treatments than current treatments without fluconazole. In other words, combination therapy with fluconazole plus pyrimethamine reduced the parasitic burden in the brain significantly, so it could be a replacement therapy in patients with intolerance sulfadiazine.

Limited Impact of the Inhibitory Receptor TIGIT on NK and T Cell Responses during Toxoplasma gondii Infection.

Resistance to the parasite Toxoplasma gondii is mediated by NK and T cell production of IFN-γ, but the failure to contract this response can lead to severe T cell-dependent immunopathology. Although the cytokines IL-10 and IL-27 prevent immune hyperactivity during toxoplasmosis, inhibitory receptors, expressed by NK and T cells, are also implicated in this process. The inhibitory receptor TIGIT is expressed on NK and T cells and competes with the costimulatory receptor CD226 for binding of the ligand CD155. During toxoplasmosis, the activation of NK and T cells is associated with increased expression of CD226 and TIGIT, whereas DCs express increased levels of CD155. To determine if the loss of TIGIT impacts NK and T cell activities, wild-type and TIGIT knockout mice were infected with T. gondii During the acute stage of infection, wild-type and TIGIT knockout mice had comparable parasite burdens and similar NK and T cell responses. Likewise, during the chronic phase of this infection, the loss of TIGIT did not affect the magnitude or phenotype of the T cell response nor the ability to control pathogen load. These data suggest that during toxoplasmosis, despite upregulation of relevant ligands, TIGIT signaling does not limit NK and T cell activities. Thus, TIGIT-independent mechanisms dominate the restraint of the immune response during toxoplasmosis.

The prevalence of Toxoplasma gondii in sheep in China: A systematic review and meta-analysis.

Toxoplasmosis is a zoonotic disease caused by Toxoplasma gondii, which is widespread in warm-blooded animals and humans. Currently, many studies regarding T. gondii infection in sheep have been published worldwide. However, there is no meta-analysis of sheep infection in China. In this study, five databases were used to retrieve articles related to T. gondii in China. A total of 59 studies from 1987 to June 30, 2020, have been included. We estimated that the combined prevalence of T. gondii in the selected period was 8.5% (3197/28,099). In the analysis of publication year, the lowest positive rate after 2010 was 7.4% (1883/17,313). Geographically, the highest prevalence of T. gondii in sheep was recorded in Southwest China 19.2% (490/2080). Analysis according to age showed that the prevalence of infection in sheep older than 12 months was significantly (P < 0.05) higher than that in younger animals. The prevalence of T. gondii in sheep farmed by farmers was 7.2% (566/6336), which was higher than 5.3% (660/3121) of large-scale farming, suggesting a role for the feeding and husbandry. We also analyzed the impact of different geographic and climatic factors on the prevalence of T. gondii infection in sheep. The results showed that the prevalence was higher in low altitude, warm, humid, and high rainfall areas. We suggest that appropriate control programs should be formulated according to the differences in reproduction patterns and geographical conditions in different regions to reduce the prevalence of T. gondii among sheep in China.

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.