Toxoplasma Gondii DNA Research Articles

Metagenomic analysis of atheroma plaques for identification of microorganisms indicates presence of Toxoplasma gondii as a possible etiological agent

BackgroundCardiovascular diseases (CVDs) are the leading cause of death worldwide. Vital organs like the heart are affected by the occlusion of blood vessels due to atherosclerotic plaque formation. However, the role of infectious agents has always been an essential subject of investigation. This study investigated the presence of microorganisms, including nanobacteria, in atherosclerotic plaques removed from human carotid arteries by microbiological and metagenomic examination. MethodsAtheroma plaque samples were obtained from 20 patients with carotid artery stenosis who had atherectomy by surgery or percutaneous intervention. Nanobacteria were grown by culturing homogenates of the atheroma plaques. Whole genome sequencing was done for samples. Because of the high percentage of Toxoplasma gondii (T. gondii) DNA, PCR investigation was applied to detect T. gondii DNA in the samples. ResultsA molecular analysis of nanobacteria revealed them to be made of human proteins, supporting the theory that they are not living organisms. According to sequencing results, samples showed that more than 50 % of the metagenomic sequences belonged to Toxoplasma gondii. PCR investigation indicated that T. gondii DNA was positive in 8 (40 %) of 20 plaques. ConclusionsFurther evidence regarding the role of T. gondii in the etiology of plaque formation may help determine the strategy for prevention and treatment of infections in preventing atheroma plaque formation in the future.

Toxoplasma gondii in rodents and shrews in Armenia, Transcaucasia

Toxoplasma gondii infections in small mammals are important because they serve as source of infection for the felids who excrete environmentally resistant oocysts in their feces. Here, the authors sought evidence for T. gondii infection in shrews and rodents in Armenia for the first time. Toxoplasma gondii DNA was detected in tissues of trapped animals using a specific PCR targeting gene with a non-coding fragment length of 529 bp. Toxoplasma gondii DNA was detected in 15 out of 137 (10.9%) samples from small mammals from 6 different localities of Armenia for the first time.

First Report on the Molecular Detection and Genetic Characterization of Toxoplasma gondii From Donkeys in Kenya.

The present study was conducted to determine the presence of Toxoplasma gondii in donkeys by molecular tests and genetic diversity analysis of the obtained DNA samples from central Kenya. A total of 363 blood samples were collected from donkeys in Meru and Kirinyaga Counties, and 96 samples that were previously seropositive for T. gondii using indirect ELISA were subjected to nested PCR based on the amplification of the internal transcribed spacer 1 (ITS-1) gene followed by DNA sequencing and phylogenetic analysis. Genotyping was performed on 15 selected positive samples using multilocus nested polymerase chain reaction restriction fragment length polymorphism (Mn-PCR-RFLP) with eight genetic markers ('SAG 2, 5'SAG 2, Alt. SAG 2, SAG 3, GRA 6, C29-2, BTUB and L358). Toxoplasma gondii DNA was detected in 36.5% (35/96) of the blood samples. The sequences obtained exhibited 98.2-99.5% homology with those deposited in GenBank. Phylogenetic analysis demonstrated that the obtained sequences are conserved and clustered with those of infecting animals from other regions of the world. Eighteen distinct T. gondii haplotypes were identified to be circulating in donkeys from central Kenya. The T. gondii DNA samples exhibited high haplotype diversity (Hd: 0.915) and limited genetic diversity (π = 0.01027). PCR-RFLP of T. gondii DNA-positive samples revealed three different genetic combinations that consisted of alleles I, II and III, indicating the dissemination of atypical genotypes. This study demonstrated that T. gondii is widespread in donkeys from Kenya and could be a possible source of infection in humans. These findings are important for designing control strategies for this parasite to improve the livestock sector, which is one of the main sources of livelihood for farmers in Kenya.

Toxoplasma gondii genotypes and frequency in domestic cats from Romania

BackgroundToxoplasma gondii is a zoonotic protozoan parasite with a heteroxenus life cycle that involves felids as the definitive hosts and any warm-blooded animal, including humans, as intermediate hosts. Cats are key players in parasite transmission as they are capable of shedding high numbers of oocysts in their feces that contaminate the environment.MethodsThe study was performed on 31 domestic cats (31.23 ± 27.18 months old) originating from rural and urban areas (5.17:1) in the center and north-west Romania. Feces (n = 31), blood (n = 28), and heart samples (n = 27) were collected. Fecal samples were analyzed by flotation technique, and PCR (529 bp repetitive element). Fecal samples with T. gondii oocysts were bioassayed in mice. Serum samples were analyzed by modified agglutination test and ImmunoComb for the detection of specific anti-T. gondii IgG antibodies. Heart samples were bioassayed in mice, and analyzed by PCR. Toxoplasma gondii positive samples were genotyped by nPCR-RFLP targeting eleven genetic loci (SAG1, SAG2, alt-SAG2, SAG3, BTUB, GRA6, c22-8, c29-2, L358, PK1, and Apico).ResultsToxoplasma gondii oocysts were found in 2 out of 31 fecal samples collected from a 3-months old stray kitten, and a 4-years old female. In total, 17 out of 27 sera were positive for T. gondii IgG antibodies. The antibody titers in MAT ranged from 1:6 to 1:384. Toxoplasma gondii DNA was detected in 7 out of 27 heart samples, and four of them were positive also by bioassay. Six T. gondii DNA samples from bioassayed mice could be assigned to ToxoDB PCR-RFLP genotype #1 or #3 (Type II) and one T. gondii DNA from heart digest to genotype #2 (Type III). Both of these genotypes are common in Europe.ConclusionsOur results revealed that the infection with T. gondii is still high in cats from Romania. The oocysts shedded by these cats represent an important source of infection for intermediate hosts, including humans. Further studies on a wider range of cases are necessary for a more exhaustive definition of the T. gondii genotypes circulating in Romania.

Oral Sex as a Potential Route for Toxoplasma Gondii Transmission: Experiment with Human Semen and Laboratory Mice Model.

Toxoplasma gondii is one of the most widespread parasites in the human population globally. Several modes of its transmission have been proposed: some are well-researched and confirmed, others remain unconfirmed. One unconfirmed hypothesis pertains to potential transmission of Toxoplasma gondii via oral sex (fellatio) in humans. A recent study found tissue cysts in the semen of men with latent toxoplasmosis. Therefore, we aimed to test the hypothesis of Toxoplasma gondii transmission through oral sex experimentally. Eighty-two laboratory mice were orally administered semen samples from 41 men with latent toxoplasmosis. These semen samples were examined for the presence of Toxoplasma gondii DNA using PCR. We detected Toxoplasma gondii DNA in three of the 41 semen samples from men with latent toxoplasmosis. Oral administration of semen samples to laboratory mice did not result in parasite transmission. We have not demonstrated the transmission of Toxoplasma to mice by oral exposure to semen from infected men. While this does not conclusively rule out the possibility of such transmission in humans, the results suggest that, if it does occur, this mode of transmission is likely infrequent.

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

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

Toxoplasma gondii Does Not Inhibit the Assisted Colonization of Eastern Barred Bandicoots (Perameles gunnii) to Phillip Island, Victoria, Australia.

Eastern barred bandicoots (Perameles gunnii) are thought to be highly susceptible to disease caused by infection with the protozoan parasite Toxoplasma gondii. This study followed a population of 67 P. gunnii introduced onto the Summerland Peninsula, Phillip Island, Australia, where the prevalence of T. gondii infection in the feral cat population was known to be very high. Prior to release, bandicoots were tested for serologic exposure to T. gondii using the modified agglutination test. A subset of bandicoots was tested on four occasions after release onto the peninsula. No seroconversion was detected at any time point. A subset of bandicoots was radiotracked after release and at two additional trapping sessions to help monitor survival. Toxoplasma gondii DNA was not detected by PCR in eight carcasses recovered for necropsy. Fourteen founder bandicoots (21% of founders) were known to be alive at 500 d post-release. A total of 29 unmarked bandicoots were trapped over the study period, confirming that the bandicoots were successfully reproducing on the island. Body weight, packed cell volume, and total plasma protein were used as measures of individual animal health; population health was inferred from these data. Body weight was significantly associated with trip number, with a general trend of increasing weight after release onto the island. This study showed that eastern barred bandicoots were able to establish a new population despite a probably high environmental load of T. gondii.

Soil-Transmitted Parasites and Non-Pathogenic Nematodes in Different Regions of Porto Alegre City, Brazil: A Comparison between Winter and Summer

We assessed the prevalence of soil-associated parasites and non-pathogenic nematodes in eight public areas of Porto Alegre (Rio Grande do Sul state, southern Brazil), the most populous city in Rio Grande do Sul. Soil samplings were carried out during the winter of 2022 and summer of 2023: A total of 80 samples were collected in winter and 80 in summer (ten samples from each sampling site per season), totaling 160 soil samples. The frequency of microscopic non-pathogenic nematode larvae was significantly higher (p = 0.048) in winter (93.75%) than in summer (82.50%). Considering the pooled data from winter and summer (n = 160) for human pathogenic parasites, the following frequencies were observed (using microscopy analysis): hookworm (filariform) larvae (1.25%), hookworm (rhabditiform) larvae (11.25%), Strongyloides spp. (filariform) larvae (0.63%), Strongyloides spp. (rhabditiform) larvae (2.5%), hookworm eggs (10.63%), Ascaris spp. eggs (10.00%), and Trichuris spp. eggs (1.25%). Hookworm (rhabditiform) larvae were the most frequent parasitic structures (15.00%) in winter, and A. lumbricoides eggs were the most frequent parasitic structures (8.75%) in summer. No statistically significant difference was observed in the frequency of pathogenic parasites between the seasons (p > 0.05). Toxoplasma gondii DNA was assessed, but all soil samples tested negative in molecular analysis. Our results indicate that soil from many regions of Porto Alegre shows a high prevalence of soil-transmitted helminths, indicating the need for improvements in social conditions and environmental sanitation in the city. Our study also suggests that climate change may affect soil biodiversity, potentially harming non-pathogenic nematodes and favoring human pathogenic parasites.

High prevalence and diversity of Toxoplasma gondii DNA in feral cat feces from coastal California.

Toxoplasma gondii is a zoonotic parasite that can cause severe morbidity and mortality in warm-blooded animals, including marine mammals such as sea otters. Free-ranging cats can shed environmentally resistant T. gondii oocysts in their feces, which are transported through rain-driven runoff from land to sea. Despite their large population sizes and ability to contribute to environmental oocyst contamination, there are limited studies on T. gondii oocyst shedding by free-ranging cats. We aimed to determine the frequency and genotypes of T. gondii oocysts shed by free-ranging domestic cats in central coastal California and evaluate whether genotypes present in feces are similar to those identified in sea otters that died from fatal toxoplasmosis. We utilized a longitudinal field study of four free-ranging cat colonies to assess oocyst shedding prevalence using microscopy and molecular testing with polymerase chain reaction (PCR). T. gondii DNA was confirmed with primers targeting the ITS1 locus and positive samples were genotyped at the B1 locus. While oocysts were not visualized using microscopy (0/404), we detected T. gondii DNA in 25.9% (94/362) of fecal samples. We genotyped 27 samples at the B1 locus and characterized 13 of these samples at one to three additional loci using multi locus sequence typing (MLST). Parasite DNA detection was significantly higher during the wet season (16.3%, 59/362) compared to the dry season (9.7%; 35/362), suggesting seasonal variation in T. gondii DNA presence in feces. High diversity of T. gondii strains was characterized at the B1 locus, including non-archetypal strains previously associated with sea otter mortalities. Free-ranging cats may thus play an important role in the transmission of virulent T. gondii genotypes that cause morbidity and mortality in marine wildlife. Management of free-ranging cat colonies could reduce environmental contamination with oocysts and subsequent T. gondii infection in endangered marine mammals and people.

Comparison of molecular techniques for the detection of Toxoplasma gondii in raw bovine milk from small rural properties in Brazil

This study aimed to research Toxoplasma gondii DNA in 102 samples of raw bovine milk from expansion tanks, in small properties located in different cities of the Midwest region of São Paulo, Brazil. For this, polymerase chain reaction (PCR) was performed with the primers TOX4/TOX5 for cPCR (conventional PCR), TgNP1/TgNP2 gene for nested PCR and the Tg18s58F/Tg18s348R for nested PCR. It was possible to detect T. gondii DNA in 18 (17.65 %) milk samples from the 102 tanks, corresponding to 4.90 % for TOX4/TOX5 primers, 12.74 % for TgNP1/TgNP2 gene and 0.98 % for Tg18s58F/Tg18s348R gene. The results showed that the TgNP1 and TgNP2 genes were more efficient to detect T. gondii DNA, and also indicated the importance of raw bovine milk as a source of human infections caused by this protozoan, being a public health problem. It is important to continue studies involving T. gondii from bovine milk considering the need for proper pasteurization, and for better comprehension regarding the epidemiology of this protozoan.

Detection of Toxoplasma gondii DNA in milk of dairy cows from southern Brazil.

Consumption of unpasteurized cow's milk may be a transmission route for some pathogenic microorganisms, but there is little information about the risk of Toxoplasma gondii infection. Blood and milk samples were collected in a paired and random fashion from 106 dairy cows and bulk-tank milk samples were also collected from each of the six farms, in southern Brazil. Serum anti-T.gondii antibodies (IgG) were detected by an indirect fluorescent antibody test (IFAT) with a cutoff point of 1:64. Nested PCR targeting the ITS1 was performed on milk samples to detect the Sarcocystidae family, confirmed to be T.gondii by Sanger sequencing. The occurrence of anti-T.gondii antibodies in the herds was 14.1%, (15/106) with seropositive cows in all herds. Antibody titers in positive samples ranged from 64 to 128. T.gondii DNA was detected in 2.8% (03/106) of the milk samples. The ITS1 sequences generated in this study were ON809793 - ON809794 and the sequencing revealed 98-100% identity with T. gondii DNA sequences deposited in GenBank. All cows PCR positive for T.gondii in milk were negative for IgG antibodies in serum, suggesting that naturally infected cows may shed T. gondii in milk in the acute phase of infection. The results of this study demonstrate that T. gondii DNA may be detected in raw cow's milk, so the potential risks of lactogenic infection should be considered. The presence of T. gondii DNA in milk does not confirm that the protozoa are viable and infective, and further investigations into the role of cow's milk in the epidemiology of toxoplasmosis are needed.

Molecular surveillance of Toxoplasma gondii in raw milk and Artisan cheese of sheep, goat, cow and water buffalo origin

This study is aimed at investigating the molecular prevalence of Toxoplasma gondii in raw milk and cheese of different animal species (sheep, goat, cow and water buffalo) in Kayseri Province, Türkiye, to provide a preliminary assessment for contamination risk. A total of 200 milk and cheese samples were analysed by real‐time PCR. Toxoplasma gondii DNA was detected in two (8%) ewes and one (4%) goat raw milk sample, while none of the cheese samples were positive. These results indicated that the presence of T. gondii DNA in raw milk samples sold in Kayseri Province might be a risk factor for public health.

Molecular Detection of Toxoplasma gondii, Neospora caninum and Encephalitozoon spp. in Vespertilionid Bats from Central Europe.

Bats may carry various viruses and bacteria which can be harmful to humans, but little is known about their role as a parasitic source with zoonotic potential. The aim of this study was to test wild bats for the presence of selected parasites: Toxoplasma gondii, Neospora caninum and microsporidia Encephalitozoon spp. In total, brain and small intestine tissues of 100 bats (52 Myotis myotis, 43 Nyctalus noctula and 5 Vespertilio murinus) were used for the DNA isolation and PCR detection of the abovementioned agents. Toxoplasma gondii DNA was detected by real-time PCR in 1% of bats (in one male of M. myotis), while all bats were negative for N. caninum DNA. Encephalitozoon spp. DNA was detected by nested PCR in 25% of bats, including three species (twenty-two M. myotis, two N. noctula and one V. murinus). Positive samples were sequenced and showed homology with the genotypes Encephalitozoon cuniculi II and Encephalitozoon hellem 2C. This is the first study on wild vespertilionid bats from Central Europe and worldwide, with a relatively high positivity of Encephalitozoon spp. detected in bats.

Foodborne protozoan parasites in fresh mussels and oysters purchased at retail in Canada

Studies worldwide have reported the presence of protozoan parasites in a variety of commercial bivalve shellfish. The uptake of these parasites by shellfish occurs during filter feeding in faecally-contaminated waters. The objective of the present study was to determine the prevalence of Giardia, Cryptosporidium and Toxoplasma in fresh, live shellfish purchased in three Canadian provinces as part of the retail surveillance activities led by FoodNet Canada (Public Health Agency of Canada). Packages containing mussels (n = 253) or oysters (n = 130) were purchased at grocery stores in FoodNet Canada sentinel sites on a biweekly basis throughout 2018 and 2019, and shipped in coolers to Health Canada for testing. A small number of packages were not tested due to insufficient quantity or poor quality. Following DNA extraction from homogenized, pooled tissues, nested PCR and DNA sequencing were used to detect parasite-specific sequences. Epifluorescence microscopy was used to confirm the presence of intact cysts and oocysts in sequence-confirmed PCR-positive samples. Giardia duodenalis DNA was present in 2.4 % of 247 packages of mussels and 4.0 % of 125 packages of oysters, while Cryptosporidium parvum DNA was present in 5.3 % of 247 packages of mussels and 7.2 % of 125 packages of oysters. Toxoplasma gondii DNA was only found in mussels in 2018 (1.6 % of 249 packages). Parasite DNA was detected in shellfish purchased in all three Canadian provinces sampled, and there was no apparent seasonal variation in prevalence. While the present study did not test for viability, parasites are known to survive for long periods in the marine environment, and these findings suggest that there is a risk of infection, especially when shellfish are consumed raw.

First description of congenital toxoplasmosis after maternal coinfection with Toxoplasma gondii and severe acute respiratory syndrome coronavirus 2: a case report

BackgroundCongenital toxoplasmosis can be associated with serious clinical consequences from fetus to adulthood. Hence, early detection is required to minimize severe sequelae through appropriate therapy. We describe the first case of a congenital toxoplasmosis after maternal coinfection with Toxoplasma gondii and severe acute respiratory syndrome coronavirus 2 and the challenging serological diagnosis of the disease in this context.Case presentationA Caucasian boy was born at 27 weeks 2 days of gestation by cesarean section due to maternal COVID-19-related respiratory failure. Postpartum serological screening of the mother revealed a previously unrecognized active Toxoplasma gondii infection. The premature child initially tested negative for anti- Toxoplasma gondii immunoglobulin A and M antibodies 1, 2 and 4 weeks after birth, whereas immunoglobulin G antibodies were only weakly positive with no evidence of child-specific production. Neither neurological nor ophthalmological abnormalities were detected. Approximately 3 months after birth, serological testing indicated a congenital toxoplasmosis by presence of immunoglobulin A and M, in combination with a child-specific immunoglobulin G synthesis. Additionally, cerebrospinal fluid was tested positive for Toxoplasma gondii DNA. Although no clinical manifestations of congenital toxoplasmosis were detected, an antiparasitic therapy was initiated to minimize the risk of late sequelae. There were no hints for a transplacental transmission of severe acute respiratory syndrome coronavirus 2.ConclusionThis case raises the awareness of possible coinfections with the risk of transplacental transmission in cases of maternal coronavirus disease 2019. The report emphasizes the need for screening vulnerable patients for toxoplasmosis in general and especially in the context of pregnancy. It becomes evident that prematurity can complicate the serological diagnosis of congenital toxoplasmosis due to a delayed antibody response. Repeated testing is recommended to carefully monitor children at risk and especially those with a history of preterm birth.

Toxoplasma gondii infection in sheep from Romania

BackgroundToxoplasmosis is a widespread zoonosis caused by the intracellular protozoan parasite Toxoplasma gondii. Limited epidemiological information is available about the prevalence of T. gondii in sheep in Romania, and a high incidence would have implications for both the economy and public health. To our knowledge, no studies are available about the T. gondii strains circulating in lambs. The objective of this study was to assess the prevalence of T. gondii in sheep (serology), lambs (serology, bioassay, PCR) and sheep abortions (PCR) in Romania. Moreover, the study aimed to perform the genetic characterization of T. gondii isolates from lambs.MethodsSerum samples collected from 2650 sheep (2067 adults and 583 lambs) were tested for anti-T. gondii antibodies (IgG) using a commercial ELISA kit. Likewise, 328 pairs of diaphragmatic muscle-serum samples were collected from lambs aged between 2 and 4 months. Lamb serum samples were analyzed using MAT for anti-T. gondii antibody detection. The diaphragm tissue samples from MAT-positive lambs (at a dilution ≥ 1:25) were bioassayed in mice. The T. gondii strains were genotyped using 15 microsatellites markers. Additionally, brain and heart samples from 76 sheep abortions were analyzed for T. gondii DNA by polymerase chain reaction (PCR) targeting the 529-bp repeat region (REP529).ResultsThe results showed that more than half of the tested sheep were T. gondii seropositive (53.5%). The seroprevalence was significantly higher in adults (61.1%) than in lambs (26.4%). The seroprevalence of T. gondii infection in slaughtered lambs, by MAT, was 37.5% (123/328). There were bioassayed in mice 56 diaphragmatic tissues from 123 seropositive lambs. Toxoplasma gondii strains were isolated from 18 (32.1%) lambs intended for human consumption. All T. gondii strains were confirmed by PCR. Six strains were genotyped using 15 microsatellite markers and belonged to genotype II. Toxoplasma gondii DNA was detected in 11.8% (9/76) of sheep abortions.ConclusionsThe present study showed the presence of T. gondii in sheep in all the regions considered in the study. The high prevalence of T. gondii infection in sheep and lambs, demonstrated by serology, molecular analysis and bioassay, highlighted that there is an important risk of human infection in consuming raw or undercooked sheep/lamb meat.Graphical

Investigation of Toxoplasma gondii and Neospora caninum in different tissues of aborted foetuses of sheep in Van Province, Türkiye: Analysis by nested PCR, histopathological and immunohistochemical methods

Toxoplasma gondii and Neospora caninum are protozoon parasites from the intracellular apicomplexan family. Toxoplasma gondii is the cause of health and economic problems in the sheep industry worldwide. Neospora caninum is usually reported in cows and leads to infections causing abortions; however, its prevalence in sheep is not clear. The present study aimed to investigate the prevalence and pathology of T. gondii and N. caninum by PCR, histopathological and immune-histochemical methods in aborted sheep foetuses collected at different sheep flocks in the Van Province, Türkiye, in 2021. Firstly, the DNA of T. gondii and N. caninum were investigated by PCR in the brain, heart, and peritoneal fluid samples from 42 sheep foetuses. Toxoplasma gondii DNA was proved in 35.7% (15/42) of foetuses whereas N. caninum DNA was not determined in any of the samples. Histopathologically, all T. gondii positive brain tissue samples showed lymphohistiocytic multifocal encephalomyelitis and additional findings included necrotizing myocarditis in the positive heart samples. Toxoplasma gondii tachyzoites were identified in the lesions (diffuse or focal mononuclear cell infiltration in the meninges, and microglia proliferation, myocarditis with oedema) by anti-T. gondii antibodies by the immunohistochemical method. Based on our results, we can conclude that T. gondii is an important agent in sheep abortions and the PCR method is a suitable method for diagnosis which can also be used in heart tissue in pathological studies.

Detection of Toxoplasma Gondii DNA in Uterine and Malignant Prostate Tissues of Cancer Patients

The current study aimed to find the Toxoplasma gondii parasite in uterine and prostate tissue models infected with cancer, which were collected from the cities of Kut and Nasiriyah, and to identify infected organs using Real-time PCR based on (B1) gene and histological examination, and the other is to identify histological changes in tissues. Cancer as a result of infection with the parasite. By examining the histological sections of samples positive for the parasite, (120) hundred and twenty tumor tissue samples fixed with formalin as well as embedded with paraffin wax were collected from incoming patients and subjected to surgical intervention for samples located in the tissue laboratories of Al-Zahra and Al-Nasiriyah Teaching Hospital and previously diagnosed with prostate and uterine cancer a month ago October 2022 to January 2023 in my province of Wasit and Dhi Qar / Iraq, whose ages ranged from (25-93 years), as it included (40) samples of the prostate for males and (80) samples of uterine tissue from females. Positive samples were taken using real-time PCR technology, and tissue sections were prepared to diagnose parasite presence and to identify histological changes in infected tissue models. The total number of samples positive for T. gondii parasite, based on the tissue sections, was 53 (44.16%) distributed according to organs, the percentage of infection was recorded in the prostate 5 (35.71%) and the uterus 19 (48.71%). These results indicated the possible role of infection with the Toxoplasma gondii parasite in causing different types of infection cancer. It is concluded that infection with the Toxoplasma gondii parasite and its presence inside the cells has a role in the development of different types of histopathological changes, which in the future turn into possible cancers in the affected organs

The Pathology of Fatal Avian Malaria Due to Plasmodium elongatum (GRW6) and Plasmodium matutinum (LINN1) Infection in New Zealand Kiwi (Apteryx spp.).

Avian malaria caused by Plasmodium species is a known cause of mortality in avifauna worldwide, however reports within New Zealand kiwi (Apteryx spp.) are scant. Postmortem reports from kiwi were obtained from the Massey University/Te Kunenga ki Pūrehuroa School of Veterinary Science Pathology Register from August 2010-August 2020. Gross lesions were described from postmortem reports, and archived H.E.-stained slides used for histological assessment. Nested PCR testing was performed on formalin-fixed paraffin-embedded tissue samples to assess the presence of Plasmodium spp. and Toxoplasma gondii DNA and cases with a PCR-positive result were sequenced to determine the lineage involved. Of 1005 postmortem reports, 23 cases of confirmed or suspected avian malaria were included in this study. The most consistent gross lesions included splenomegaly, hepatomegaly, and interstitial pneumonia with oedema. Histological lesions were characterised by severe interstitial pneumonia, pulmonary oedema, interstitial myocarditis, hepatic sinusoidal congestion and hypercellularity, and splenic macrophage hyperplasia and hyperaemia/congestion with numerous haemosiderophages. Cytoplasmic meronts were consistently found within endothelial cells of a variety of tissues, and within tissue macrophages of the liver, lung and spleen. A diagnosis of avian malaria was confirmed via PCR testing in 13 cases, with sequencing revealing P. matutinum (LINN1) and P. elongatum (GRW6) as the species involved. This is the largest case series describing the pathology of avian malaria as a cause of mortality in endemic New Zealand avifauna.

Seroprevalence and genetic characterization of Toxoplasma gondii in domestic pigs intended for human consumption in Cuba.

Domestic pigs are considered as one of the main intermediate hosts in the zoonotic transmission of Toxoplasma gondii in many countries. Serological and molecular studies are warranted to better understand the epidemiology and transmission patterns of this parasite worldwide. To date, seroepidemiological information on T.gondii in domestic pigs in Cuba is very scarce and there are no reports of T.gondii genotypes circulating in this country. Here, we aimed to estimate the seroprevalence of T.gondii and provide genetic characterization of the strains circulating in slaughtered pigs intended for human consumption in Central Cuba. Seroprevalence was determined in 450 serum samples from slaughtered pigs in Villa Clara province using ELISA. Anti-Toxoplasma gondii IgG antibodies were detected in 100 animals (22.2%, 95% CI: 18.5-26.2). Conventional PCR of the 529-bp marker of T.gondii was performed in hearts and diaphragm tissues of all ELISA-seropositive pigs. Toxoplasma gondii DNA was detected in four animals. Further genetic characterization of the positive DNA samples was performed by multilocus PCR-RFLP and PCR-sequencing typing tools. Molecular analysis revealed four different genetic profiles that were combinations of type I, II, III and u-1 alleles, suggesting the circulation of non-clonal genotypes of T.gondii in domestic pigs in Cuba. Our results indicate that T.gondii is widely distributed in slaughtered pigs in this country, which might have important implications for public health. To the best of our knowledge, this is the first report on genetic characterization of T.gondii in Cuba. Although preliminary, the results suggest a high genetic diversity of T.gondii in the study region. Further studies based on parasite isolation are needed to definitively identify the genotypes circulating and characterize the virulence of strains detected in pigs in Cuba, and to assess the risk of zoonotic transmission from pork products in this country.