Genetic characterization of Toxoplasma gondii from human and chicken isolates from Argentina.

This study aimed to determine the nPCR-RFLP genotypes of newly obtained T. gondii isolates from human congenital toxoplasmosis cases in Argentina and to determine their allelic profiles for virulence genes ROP18/ROP5. In addition, the ROP18/ROP5 profiles were also determined for previously characterized T. gondii samples. Isolation from congenital toxoplasmosis cases was carried out in mouse bioassay from two placentas (P1 and P2). Genotyping for the new human isolates was performed by nPCR-RFLP using 10 markers. The samples analyzed for ROP18/ROP5 included the two newly obtained isolates (from the congenital toxoplasmosis cases) and nine previously genotyped T. gondii DNA samples from humans and chickens. The results for P1 and P2 named as TgHm18-02Arg and TgHm19-01Arg showed ToxoDB genotypes #14 (non-archetypal) and #2 (clonal type III), respectively. Non-archetypal #14 has been isolated from human cases before in Argentina. However, this is the first report of T. gondii clonal type III in a human case in the country. The ROP18/ROP5 combination was detected in nine samples: 3/3 (n = 1), 4/3 (n = 4), 4/4 (n = 3), and 3-4/4 (n = 1). Notably, the 4/4 profile was identified for the first time and exclusively in T. gondii samples from Misiones province (which borders southern Brazil). Further studies are required to corroborate the regionalization of the ROP18/ROP5 profiles in Argentina.

Clinical outcome and vertical transmission variability among canine Neospora caninum isolates in a pregnant mouse model of infection.

We compared the clinical outcome and vertical transmission of six canine Neospora caninum isolates using a pregnant BALB/c model. Four of the isolates were obtained from oocysts of naturally infected dogs (Nc-Ger2, Nc-Ger3, Nc-Ger6 and Nc-6Arg) and two were from diseased dogs with neurological signs (Nc-Bahia and Nc-Liv). The dams were inoculated with 2×106 tachyzoites of each isolate at day 7 of pregnancy. Morbidity, mortality and the antibody responses were evaluated in both the dams and the offspring, as was parasite transmission to the progeny. The mortality rates varied from 100% in Nc-Bahia and Nc-Liv-infected pups to 19% or less for those infected with the isolates from oocysts. The vertical transmission rates varied from 9 to 53% for N. caninum from oocysts, compared with 100% for the Nc-Liv and Nc-Bahia isolates. All dams showed specific IgG responses against tachyzoite and rNc-GRA7 antigens, confirming Neospora infection. The highest IgG levels were detected in mice inoculated with the Nc-Liv and Nc-Bahia isolates. These results demonstrate marked differences in virulence between the N. caninum isolates obtained from oocysts and neurologically affected dogs. This variability could help us to explain the differences in the outcome of the infection in definitive and intermediate hosts.

A 20-year serological survey of Toxoplasma gondii and Neospora caninum infection in dogs with neuromuscular disorders from urban areas in Argentina.

Toxoplasma gondii and Neospora caninum infections may be associated with neuromuscular disorders in dogs. The aim of this study was to assess the seroprevalence to these protozoan parasites in dogs with neuromuscular disease from urban areas of Buenos Aires province, Argentina, over a period of 20 years, and to evaluate the association of seropositivity and antibody titres with different variables such as sex, breed and age. For this, a total of 7238 serum samples from urban owned dogs were analysed by the indirect fluorescence antibody test (IFAT) for specific IgG antibodies. The observed seropositivity rates were 35.7 % for T. gondii and 25.7 % for N. caninum. Crossbred dogs had a significantly higher seroprevalence for T. gondii than purebred dogs (41 % vs. 29.3 %), while a trend towards significance was observed for N. caninum, which was slightly higher in purebred dogs (26 % vs. 23.6 %). Seroprevalence for both parasites increased with age and was higher in older animals. Regarding the distribution of specific antibody titres, the most frequent IFAT T. gondii titre found was 100 and for N. caninum it was ≥800. For toxoplasmosis, there was no association with age group, and low titres (50, 100 and 200) predominated in all groups. However, for neosporosis, age and titres were significantly associated for one age group, with dogs under 12 months of age having a higher proportion of high titres (400 and 800). The trend in the seroprevalence for T. gondii is increasing over the years and lower antibody titres predominate in the dogs studied, which may be more related to the presence of chronic infections and not necessarily to the clinical signs of the animals. Despite the generally low titres observed for toxoplasmosis in this study, it is important to highlight the high seroprevalence found in our region, as dogs can act as sentinels of environmental contamination and as indicators of possible human infection. In the case of neosporosis, although the trend in seroprevalence in dogs with signs appears to be decreasing over the years, our work shows that higher antibody titres predominate, and are probably related to the clinical signs presented by the dogs. This study provides the most recent epidemiological data and serological profiles of T. gondii and N. caninum infections in a large number of canine sera from urban areas in Argentina, providing relevant information for clinical veterinarians and epidemiologists in order to understand the circulation of the parasites.

[Argentine consensus of congenital toxoplasmosis]. / Consenso argentino de toxoplasmosis congénita 2020.

Mother-to-child transmission in Toxoplasma gondii infection occurs only when the infection is acquired for the first time during pregnancy. Diagnosis of maternal infection and the newborn is achieved by a combination of serological tests, clinical features and ultrasound images. An early diagnosis of maternal infection allows treatment that offers a reduction both in transmission rate and risk of congenital damage. The aim of this expert consensus was to review the scientific literature which would enable an update of the clinical practice guideline of prevention, diagnosis and treatment of congenital toxoplasmosis in our country.

La transmisión vertical de la infección por Toxoplasma gondii ocurre cuando la madre se infecta por primera vez en el transcurso del embarazo. El diagnóstico de la infección materna y la del recién nacido se logra con el conjunto de pruebas serológicas, hallazgos clínicos y ecográficos. El reconocimiento temprano de la infección materna permite un tratamiento que reduce la tasa de transmisión y el riesgo de daño en el producto de la concepción. El objetivo de este consenso de expertos fue revisar la literatura científica para actualizar las recomendaciones de práctica clínica respecto de la prevención, el diagnóstico y el tratamiento de la toxoplasmosis congénita en nuestro país.

Isolation of Neospora caninum from a beef cattle fetus from Argentina: Immunopathological and molecular studies.

Neospora caninum is an important abortifacient agent affecting mainly cattle worldwide. The aim of the present work was to describe the histopathological findings in a naturally infected beef cow and its midterm fetus caused by a genetically defined N. caninum isolate in Argentina. A N. caninum seropositive multiparous Aberdeen Angus pregnant cow and its fetus in the sixth month of gestation were submitted for histopathological, immunohistochemical, serological, and molecular studies and parasite isolation. The cow belonged to a beef herd under extensive management, with a N. caninum seroprevalence of 11%, and low level of annual abortion rate (≤ 5%). The dam had mild lymphocytic infiltrate in CNS, heart and uterus and no parasites were detected by Immunohistochemistry (IHC). No parasitic DNA was detected in the dam's brain, and gamma interferon knockout mice inoculated with brain material did not become infected. Clusters of tachyzoites and parasitic DNA were detected in the placenta by IHC and PCR, respectively. However, isolation from the placenta was unsuccessful. The fetus developed specific antibodies and an inflammatory response was detected in multiple organs. Furthermore, in vitro and in vivo isolation was achieved from gamma interferon knockout mice inoculated with CNS from the fetus. Multilocus-microsatellite typing revealed a genetically defined N. caninum isolate similar to the previously reported as MLG 72. We report the first N. caninum isolate from beef cattle in Argentina.

Toxoplasma gondii and Neospora caninum infections in synanthropic rodents from Argentina.

Toxoplasma gondii and Neospora caninum are closely related coccidian parasites (phylum Apicomplexa). This is the first study from urban synanthropic rodent species that involved serological and molecular diagnosis of T. gondii and N. caninum infection, and genotyping of T. gondii in Argentina. A total of 127 rodent samples were trap captured: Mus musculus (n = 78), Rattus norvegicus (n = 26) and Rattus rattus (n = 23). Antibodies against T. gondii and N. caninum were detected by IFAT in 32.8% (40/122) and 0.8% (1/122) of rodent samples, respectively, demonstrating contact with these protozoans. Additionally, T. gondii DNA was detected in 3.3% (4/123) of rodent central nervous system samples and 2 samples were genotyped by multilocus nPCR-RFLP. Neospora caninum DNA was not detected by PCR. The 2 genotyped samples were type III allele for all markers except for SAG-1 (type I for Rat1Arg and type II/III for Rat2Arg) and were identified as #48 and #2 (likely) according to the allele combinations reported on Toxo DB (Toxo-DB). The results of the present study revealed a wide distribution of T. gondii and less for N. caninum, in synanthropic rats and mice in the studied area.

Toxoplasma gondii and Trichinella infections in wild boars (Sus scrofa) from Northeastern Patagonia, Argentina.

Wild boar (Sus scrofa) was introduced in many countries of the world and is recognized as carrier of many infectious diseases. Wild game meat consumption is recognized as a source of transmission of Toxoplasma gondii and Trichinella spp. The aim of the present study was to evaluate the prevalence of antibodies to T. gondii and Trichinella spp. in free-range wild boars in Northeastern Argentine Patagonia. Between 2014 and 2018, 144 blood samples and 423 muscle samples from 423 carcasses were collected. To detect T. gondii IgG, 144 sera were processed by an immunofluorescent antibody test, and to detect anti-Trichinella IgG, 125 sera and 304 muscle juice samples were processed by ELISA. Detection of first stage larvae in muscle was performed by artificial digestion. A total of 423 wild boars muscle samples were negative to Trichinella spp. by artificial digestion. Antibodies to Trichinella spp. were detected in 2.4% (3/125) of serum samples and in 1.64% (5/304) of meat juice samples. Antibodies to T. gondii infection were detected in 12.5% (18/144) of the serum samples. This is the first study to reveal the presence of antibodies to T. gondii in wild boars from Argentina. The present results suggest that consumption of raw or undercooked wild boar meat could represent a potential source risk for toxoplasmosis in humans and that Trichinella spp. is infrequent and/or that it circulates in low burdens among wild boars in Northeastern Patagonia.

Fatal canine encephalitozoonosis in Latin America, first report.

Encephalitozoon cuniculi is an obligate, intracellular microsporidian organism capable of establish infection in a wide variety of animals. In carnivores it may cause a sporadic, severe disease in the first few months of life, which usually culminates with the death of the animal. The objective of this study was to report a natural fatal case of encephalitozoonosis in a puppy from Argentina. Clinical signs included reduced appetite, depression, vocalizing, weight loss, weakness, convulsions and recumbency. No significant gross lesions were noticed at necropsy. Microscopically, severe, diffuse, lymphocytic encephalitis was seen. Large cytoplasmic vacuoles containing spores, morphologically compatible with E. cuniculi, were present within endothelial cells of brain and kidney, in renal tubular epithelium and hepatocytes. Encephalitozoon cuniculi DNA was detected by PCR in the kidney. Antibody titers to E. cuniculi in serum from the surviving puppies and the dam were ≥1:200. This report contributes to our understanding of neurologic disease in puppies. Encephalitozoonosis should be considered as a differential diagnosis in cases of fatal encephalitis in puppies.

Microsatellite pattern analysis of Neospora caninum from a naturally infected goat fetus.

Neospora caninum is an apicomplexan protozoan responsible for abortion in ruminants. The present study aimed to diagnose an abortion from an Anglo Nubian goat from a dairy herd located in Buenos Aires province, Argentina. The goat delivered a fetus of approximately 3 months gestation that was studied by indirect fluorescent antibody test (IFAT), Immunoblot (IB), histopathology (HP), immunohistochemistry (IHC), and molecular assays (PCR, sequencing and microsatellite genotyping). Interferon gamma knock-out mice were inoculated with a pool of tissues for isolation attempts. The mother had IFAT titers of 1:3200 and 1:400 for N. caninum and Toxoplasma gondii, respectively, as well as positive IB reactions, whereas the fetus was seronegative to both parasites by IFAT and IB. The fetus had severe multifocal necrotizing myocarditis and hepatitis, moderate interstitial pneumonia, and nephritis. Myocardium sample resulted positive by IHC, evidencing clusters of N. caninum tachyzoites within myocardiocytes associated with histopathological lesions. Neospora caninum-DNA was detected by PCR in heart, liver, lungs, kidney, and muscle from the fetus, and was negative for T. gondii by PCR. NC-5 and 18 S rRNA gene fragment sequences showed 100% identity with N. caninum. Inoculated mice bled 30 days post-inoculation resulted seronegative to N. caninum and T. gondii by IFAT, and showed no clinical signs. Multilocus-microsatellite genotyping revealed a genetic profile that differed from previously reported N. caninum genotypes, with unique MS21 and MS10 alleles. These findings indicate that N. caninum was efficiently transmitted from the mother to the fetus. We report the first case of direct detection of N. caninum in a goat fetus in Argentina and N. caninum microsatellite genotyping in naturally infected goat.

Seroprevalence of Toxoplasma gondii and Neospora caninum infections in goats from two Argentinean provinces.

The aim of this study was to compare the seroprevalence of Toxoplasma gondii and Neospora caninum in goats from two Argentinean provinces raised under different management conditions. A total of 2922 serum samples from adult goats of Córdoba (n=2187) and Buenos Aires provinces (n= 735), Argentina, were assayed by indirect fluorescence antibody test (IFAT, cut-off 1:100) for antibodies to T. gondii and N. caninum. Seroprevalence was 40.8% (CI 39.0%-42.6%) and 5.5% (CI 4.7%-6.4%) for T. gondii and N. caninum, respectively. The seropositivity for both infections was higher in goats from dairy farms, resulting as follows: for T. gondii 32.7% (CI 30.8%-34.8%) in extensive farms and 59.3% (CI 56.1%-62.6%) in dairy farms and for N. caninum 4.1% (CI 3.2%-4.9%) in extensive farms and 8.8% (CI 6.9%-10.7%) in dairy farms. This is the first extensive seroepidemiology investigation for T. gondii and N. caninum in goats in Argentina.

Consenso Argentino de toxoplasmosis congénita 2020 / Argentine Consensus of congenital toxoplasmosis

Resumen La transmisión vertical de la infección por Toxoplasma gondii ocurre cuando la madre se infecta por primera vez en el transcurso del embarazo. El diagnóstico de la infección materna y la del re cién nacido se logra con el conjunto de pruebas serológicas, hallazgos clínicos y ecográficos. El reconocimiento temprano de la infección materna permite un tratamiento que reduce la tasa de transmisión y el riesgo de daño en el producto de la concepción. El objetivo de este consenso de expertos fue revisar la literatura científica para actualizar las recomendaciones de práctica clínica respecto de la prevención, el diagnóstico y el tratamiento de la toxoplasmosis congénita en nuestro país.

Abstract Mother-to-child transmission in Toxoplasma gondii infection occurs only when the infection is acquired for the first time during pregnancy. Diag nosis of maternal infection and the newborn is achieved by a combination of serological tests, clinical features and ultrasound images. An early diagnosis of maternal infection allows treatment that offers a reduction both in transmission rate and risk of congenital damage. The aim of this expert consensus was to review the scientific literature which would enable an update of the clinical practice guideline of prevention, diagnosis and treatment of congenital toxoplasmosis in our country.

Seroprevalence of Toxoplasma gondii in sows from slaughterhouses and in pigs from an indoor and an outdoor farm in Argentina.

The objective of the present study was to determine the prevalence of Toxoplasma gondii antibodies from slaughter sows and from pigs raised at an indoor and an outdoor swine farm. Serum samples were obtained from 230 slaughter sows belonging to 83 farms distributed in 5 provinces. Blood samples were collected monthly from pigs of different ages from an intensive management indoor farm (farm 1). A cross-sectional study was carried-out from an outdoor farm (farm 2). All sera were tested for T. gondii antibodies by the modified agglutination test (MAT), using formalin-fixed tachyzoites as antigen. An antibody titer > or =1:25 was considered positive. Antibodies to T. gondii were detected in 87 (37.8%) of 230 sows sera. Distribution among provinces was: 37.1% from Santa Fe, 62.8% from Buenos Aires, 3.3% from San Luis, 58.7% from La Pampa and 24% from Córdoba. Four of 88 (4.5%) serum samples from farm 1 had antibodies to T. gondii and none of the negative pigs seroconverted. However, 45 of 112 samples from farm 2 were positive (40.2%) with the following distribution: sows 100%; nursery 40%; growers 13.8% and fatteners 20%. It is concluded that the prevalence of T.gondii antibodies among sows seems to be quite variable. T. gondii prevalence was related to the facilities and management of the farm.

Seroepidemiology of beef and dairy herds and fetal study of Neospora caninum in Argentina.

The purpose of the present work was to study the epidemiology of Neospora caninum in beef and dairy herds in the Humid Pampas of Argentina. The seroprevalence of N. caninum was evaluated in 2414 serum samples of cows from beef and dairy farms. An indirect fluorescent antibody test (IFAT) was used to determine specific antibodies. The sera was screened at a dilution >or=1:200 and >or=1:600 in cows with reproductive disease antecedents and without them, respectively. Cows without history of reproductive diseases from nine beef and fifteen dairy farms were grouped according to the percentage (> or or

Neospora caninum is a cause of perinatal mortality in axis deer (Axis axis).

Neospora caninum is a worldwide distributed protozoan that may cause neuromuscular disease in dogs and reproductive failure in domestic and wild ruminants. One axis fawn (Axis axis) and four neonates from the same deer herd died at a zoo in Argentina within a four-month period. The fawn presented with dilatation of the anal sphincter at birth and incontinence, developed weakness and ataxia and died at 14 days of age. At necropsy, a mega formation of the distal large intestine was observed. Microscopically, non-suppurative encephalitis, suppurative bronchopneumonia, fibrin necrotic enteritis and degenerative changes in the liver were observed in hematoxilin and eosin-stained tissue sections, and thick-walled N. caninum-like cysts were observed in fresh brain samples. Serologic studies for N. caninum revealed an IFAT titer of 1:6400 in the fawn and 1:25, 1:400, 1:3200 and 1:6400 in the neonates. N. caninum DNA was detected in brain samples from the fawn and from one neonate by PCR, and the parasite was isolated in vitro from the fawn' brain after passage through gerbils (Meriones unguiculatus) and gamma-interferon knock-out mice. N. caninum DNA obtained from the fawn, neonate and isolated parasites showed the same microsatellite pattern. This suggests a common infection source for both animals. The diagnosis of N. caninum infection was confirmed, suggesting its association with perinatal mortality in captive axis deer. To the best of our knowledge, this is the first report of clinical disease associated to N. caninum infection in axis deer and of isolation of the parasite from this wild ruminant species.

Toxoplasma gondii and Neospora caninum infections in synanthropic rodents from Argentina / Infecções por Toxoplasma gondii e Neospora caninum em roedores sinantrópicos da Argentina

Abstract Toxoplasma gondii and Neospora caninum are closely related coccidian parasites (phylum Apicomplexa). This is the first study from urban synanthropic rodent species that involved serological and molecular diagnosis of T. gondii and N. caninum infection, and genotyping of T. gondii in Argentina. A total of 127 rodent samples were trap captured: Mus musculus (n = 78), Rattus norvegicus (n = 26) and Rattus rattus (n = 23). Antibodies against T. gondii and N. caninum were detected by IFAT in 32.8% (40/122) and 0.8% (1/122) of rodent samples, respectively, demonstrating contact with these protozoans. Additionally, T. gondii DNA was detected in 3.3% (4/123) of rodent central nervous system samples and 2 samples were genotyped by multilocus nPCR-RFLP. Neospora caninum DNA was not detected by PCR. The 2 genotyped samples were type III allele for all markers except for SAG-1 (type I for Rat1Arg and type II/III for Rat2Arg) and were identified as #48 and #2 (likely) according to the allele combinations reported on Toxo DB (Toxo-DB). The results of the present study revealed a wide distribution of T. gondii and less for N. caninum, in synanthropic rats and mice in the studied area.

Resumo Toxoplasma gondii e Neospora caninum são parasitas coccídeos intimamente relacionados (filo Apicomplexa). Este é o primeiro estudo de espécies de roedores sinantrópicos urbanos, o qual envolveu diagnósticos sorológicos e moleculares da infecção por T. gondii e N. caninum e genotipagem de T. gondii na Argentina. Um total de 127 amostras de roedores foram obtidas: Mus musculus (n = 78), Rattus norvegicus (n = 26) e Rattus rattus (n = 23). Anticorpos contra T. gondii e N. caninum foram detectados pela IFAT em 32,8% (40/122) e 0,8% (1/122) das amostras de roedores, respectivamente, demonstrando contato com esses protozoários. Adicionalmente, o DNA de T. gondii foi detectado em 3,3% (4/123) das amostras do sistema nervoso central de roedores e duas amostras foram genotipadas por nPCR-RFLP multilocus. O DNA de N. caninum não foi detectado por PCR. As 2 amostras genotipadas eram do tipo III para todos os marcadores, exceto para SAG-1 (tipo I para Rat1Arg e tipo II / III para Rat2Arg) e foram identificadas como # 48 e # 2 (provavelmente) de acordo com as combinações de alelos relatadas no Toxo DB (Toxo-DB). Os resultados do presente estudo indicam uma ampla distribuição de T. gondii e menor para N. caninum , em ratos e camundongos sinantrópicos na área estudada.