RESUMEN
Toxoplasma gondii causes severe disease in congenitally infected fetuses. The severity of fetal infection is related to the gestational stage at the time of maternal infection, parasite burden, and genotypic characteristics. South America has a high incidence of congenital toxoplasmosis and has the highest genotypic diversity of the parasite. In Brazil, clinical toxoplasmosis in children is notorious, however there are very limited data regarding the strains recovered from congenital infections. In this study, T. gondii strains from two cases of severe congenital toxoplasmosis from the São Paulo metropolitan area were isolated (TgHumIMTBr2 and TgHumIMTBr3) and biologically and molecularly characterized using polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) and microsatellite analysis, revealing a new non-archetypal virulent genotype designated as #318. The other isolate, genotype #175, has already been described in domestic and wild animals in Brazil, but is now associated with acute toxoplasmosis in humans. These data reinforce the role of non-archetypal T. gondii genotypes in the severity of human congenital toxoplasmosis, highlighting the importance of studies focused on parasite isolation and genotyping for a better understanding of the virulence of isolates from human toxoplasmosis and contributing to the knowledge of the diversity of T. gondii in Brazil.
Asunto(s)
Toxoplasma , Toxoplasmosis Congénita , Brasil/epidemiología , Niño , Variación Genética , Genotipo , Humanos , Polimorfismo de Longitud del Fragmento de Restricción , Toxoplasma/genética , Toxoplasmosis Congénita/epidemiología , Toxoplasmosis Congénita/parasitologíaRESUMEN
BACKGROUND: Toxoplasmosis is a zoonotic disease that affects humans and warm-blooded animals. This study describes an outbreak of toxoplasmosis in howler monkeys (Alouatta sp.) and survival of capuchins (Sapajus apella), under the same environmental conditions. METHODS: Howler monkeys were submitted to post-mortem examination. Tissue samples were processed to histopathology and immunohistochemistry to detect lesions and tachyzoites of Toxoplasma gondii. Tissue samples were also frozen and submitted to PCR and genotyping of T. gondii. RESULTS: Typical lesions were observed in several organs including the liver, lymph node, and brain, with intralesional cysts and tachyzoites of T. gondii demonstrated by immunohistochemistry. T. gondii genomic sequences were amplified by PCR, and genotyping characterized the same T. gondii clone in all howler monkeys. CONCLUSIONS: Our results support the notion that some species of neotropical primates are highly susceptible to toxoplasmosis and the hypothesis that capuchins (S. apella) may be resistant.
Asunto(s)
Alouatta , Genotipo , Enfermedades de los Monos/parasitología , Toxoplasma/genética , Toxoplasmosis Animal/parasitología , Animales , Animales de Zoológico , Brasil/epidemiología , Femenino , Masculino , Enfermedades de los Monos/epidemiología , Sapajus apella/parasitología , Sapajus apella/fisiología , Toxoplasmosis Animal/epidemiologíaRESUMEN
Pulmonary toxoplasmosis is rare in immunocompetent patients. Herein, a Toxoplasma gondii strain isolated in Brazil from an immunocompetent patient who had severe pulmonary involvement was biologically and molecularly characterized for the first time. The TgHumIMTBr1 isolate was bioassayed in mice showing a virulent phenotype. Restriction fragment length polymorphism (RFLP) genotyping using 11 markers [SAG1, SAG2 (5´3´SAG2 and alt. SAG2), SAG3, BTUB, GRA6, c22-8, c29-2, L358, PK1, Apico and CS3] revealed a new non-archetypal genotype assigned as #312. Genotyping using ROP18/ROP5 markers exhibited the virulent combination of alleles 4 and 1. Microsatellite analysis using 15 markers (TUB2, W35, TgM-A, B18, B17, M33, IV.1, X1.1, N60, N82, AA, N61, N83, M48 and M102) revealed an atypical genotype with three unique alleles and a rare combination of alleles 246 (W35) and 203 (TgM-A) that is typical of the Amazon region. Non-archetypal genotypes with unique alleles may function in the occurrence of severe toxoplasmosis in immunocompetent patients in Brazil. Attempts to isolate or molecularly detect T. gondii for further genotyping studies would contribute to the understanding of causes related to the severity of toxoplasmosis in immunocompetent patients.
Asunto(s)
Enfermedades Pulmonares Parasitarias/parasitología , Toxoplasma/clasificación , Toxoplasmosis/parasitología , Adulto , Alelos , Animales , Brasil , Variación Genética , Genotipo , Humanos , Inmunocompetencia , Masculino , Ratones , Filogenia , Reacción en Cadena de la Polimerasa , Polimorfismo de Longitud del Fragmento de Restricción , Toxoplasma/genética , Toxoplasma/aislamiento & purificaciónRESUMEN
The present study evaluated the distribution and viability of Toxoplasma gondii tissue cysts in the organs and Brazilian commercial cuts of experimentally infected pigs. The pigs were infected with 3 × 103 oocysts of the T. gondii isolate TgCkBr57 (Type BrII). Mouse bioassays were performed on the brain, retina, tongue, diaphragm, and heart as well as the following muscle cuts: loin (longissimus), coppa (longissimus, spinalis dorsi, rhomboideus), tenderloin (psoas major), outside flat (biceps femoris), topside (semimembranosus), and top sirloin (gluteus medius). Toxoplasma gondii was isolated from the coppa, heart, diaphragm, and tongue of three pigs; from the tenderloin, outside flat, and brain of two pigs; and from the top sirloin and loin of one pig. Thus, the viability of T. gondii cysts was observed in all of the organs and cuts evaluated (except for the topside and retina), demonstrating the broad distribution of this parasite in pig organs and commercial meat cuts, and the importance of this species as a source of human infection.
Asunto(s)
Carne/parasitología , Enfermedades de los Porcinos/parasitología , Porcinos/parasitología , Toxoplasma/aislamiento & purificación , Toxoplasmosis Animal/parasitología , Animales , Encéfalo/parasitología , Brasil , Diafragma/parasitología , Femenino , Corazón/parasitología , Humanos , Ratones , Oocistos/aislamiento & purificación , Retina/parasitología , Lengua/parasitologíaRESUMEN
Toxoplasmosis is a disease with a worldwide distribution that affects a wide variety of animal species, though with rare descriptions in chickens. We describe the clinical, epidemiological, pathological, and molecular aspects of a toxoplasmosis outbreak in domestic chickens and guinea fowl in southern Brazil. The flock was composed of 47 domestic chickens and 29 guinea fowl. Of these, 22 birds showed clinical signs of lethargy, anorexia, and neurological signs over a clinical course of 24-72 h, and 15 died. Epidemiological data were obtained through fieldwork performed at the chicken farm and necropsies of six birds. Gross lesions were absent at necropsy, and histopathological findings included inflammatory infiltrate of macrophages, lymphocytes, and plasma cells and necrosis in several tissues associated with intralesional Toxoplasma gondii. Immunohistochemistry for T. gondii was positive. Additionally, restriction fragment length polymorphism (RFLP) analysis with 11 markers (SAG1, SAG2 (5'3'SAG2 and alt. SAG2), SAG3, BTUB, GRA6, c22-8, c29-2, L358, PK1, Apico, and CS3) and microsatellite (MS) analysis with 15 markers (TUB2, W35, TgMA, B18, B17, M33, IV.1, XI.1, N60, N82, AA, N61, N83, M48, and M102) were performed. PCR-RFLP revealed T. gondii genotype ToxoDB-PCR-RFLP #280, and MS analysis also showed a unique genotype. This is the first description of this genotype in chickens and adds to the evidence suggesting considerable genotypic diversity of T. gondii in Brazil.
Asunto(s)
Brotes de Enfermedades/veterinaria , Galliformes/parasitología , Enfermedades de las Aves de Corral/patología , Toxoplasma/aislamiento & purificación , Toxoplasmosis Animal/patología , Animales , Brasil/epidemiología , Pollos/parasitología , Genotipo , Reacción en Cadena de la Polimerasa , Polimorfismo de Longitud del Fragmento de Restricción , Enfermedades de las Aves de Corral/epidemiología , Enfermedades de las Aves de Corral/parasitología , Toxoplasma/genética , Toxoplasmosis Animal/epidemiología , Toxoplasmosis Animal/parasitologíaRESUMEN
Toxoplasmosis is a zoonotic disease caused by the protozoan Toxoplasma gondii. Infections occur via the ingestion of oocysts, consumption of cysts containing bradyzoites, and transplacental transmission of tachyzoites. Diversity in T. gondii strains may affect the outcome of clinical toxoplasmosis. The consumption of horse meat is a common practice in some parts of the world. The objectives of the present study were to isolate and genotype T. gondii from horses from an abattoir in the state of Rio Grande do Sul in southern Brazil that exports horse meat to Europe. Antibodies to T. gondii were found in 32.5% (13/40) of the horses using the modified agglutination test (MAT) with a cut-off of 1:25. Tissues from the 13 seropositive horses were bioassayed in mice, and one isolate, designated TgHorseBrRS1, was obtained. PCR-RFLP of the isolate revealed the ToxoDB-RFLP #228 genotype, a typical non-archetypal Brazilian genotype, and microsatellite analysis showed a unique non-archetypal genotype. This study showed that horses from Brazil can harbor viable T. gondii in their tissues, suggesting that recommendations to consumers should be made, especially in European countries where consumption of raw horse meat is common.
Asunto(s)
Enfermedades de los Caballos/parasitología , Carne/parasitología , Toxoplasma/aislamiento & purificación , Toxoplasmosis Animal/parasitología , Mataderos/estadística & datos numéricos , Animales , Bioensayo , Brasil , Europa (Continente) , Inocuidad de los Alimentos , Genotipo , Caballos , Humanos , Ratones , Oocistos/clasificación , Oocistos/genética , Oocistos/aislamiento & purificación , Polimorfismo de Longitud del Fragmento de Restricción , Toxoplasma/clasificación , Toxoplasma/genéticaRESUMEN
The biological and genetic diversity of Neospora caninum is very limited because of availability of only a few viable isolates worldwide. This study describes the isolation and biological and molecular characterization of a new viable isolate of N. caninum (NC-SP1), from a cattle in Brazil. Approximately 400 g of brain from a naturally infected adult male cattle from an abattoir was fed to a 2-month-old dog. Neospora-like oocysts were observed on day 7 post-inoculation (PI) and the duration of oocyst shedding was 14 days. The DNA obtained from oocysts was characterized molecularly and the final sequence was 99% identical to homologous sequences of N. caninum available in GenBank®. For bioassay, gerbils (Meriones unguiculatus) were orally inoculated with 10 100 and 1000 oocysts; all gerbils remained clinically normal but developed N. caninum antibodies 14 days PI. Cell culture isolation was successful using the brain homogenate from one of the gerbils and tachyzoites were observed 24 days PI. Microsatellite genotyping revealed a unique genetic profile for this new reference isolate.
Asunto(s)
Enfermedades de los Bovinos/parasitología , Coccidiosis/veterinaria , Neospora/aislamiento & purificación , Animales , Anticuerpos Antiprotozoarios/sangre , Bioensayo/veterinaria , Encéfalo/parasitología , Brasil , Bovinos , Coccidiosis/parasitología , ADN Protozoario/química , Perros , Heces/parasitología , Femenino , Técnica del Anticuerpo Fluorescente Indirecta/veterinaria , Técnicas de Genotipaje/veterinaria , Gerbillinae , Masculino , Repeticiones de Microsatélite , Neospora/genética , Neospora/inmunología , Oocistos/genética , Oocistos/inmunología , Oocistos/aislamiento & purificación , Reacción en Cadena de la Polimerasa/veterinaria , Polimorfismo de Longitud del Fragmento de Restricción , Suero/parasitologíaRESUMEN
Toxoplasmosis is a widely distributed disease that infects birds and mammals, including humans. Acute clinical course of toxoplasmosis is considered to be rare among domestic rabbits (Oryctolagus cuniculus). The aim of this study was to present the first report of fatal acute disease caused by Toxoplasma gondii type BrIII genotype, a typical Brazilian clonal lineage, in a domestic rabbit. T. gondii was identified in histological sections of spleen and liver tissue, and these tissues were also immunohistochemically positive for T. gondii. After the histopathological and immunohistochemical confirmation of T. gondii, the genotype of this pathogen was determined via PCR-RFLP with 11 markers (SAG1, SAG2, SAG3, BTUB, GRA6, c22-8, c29-2, L358, PK1, Apico, and CS3) and via microsatellite (MS) analysis with 15 markers (TUB2, W35, TgMA, B18, B17, M33, IV.1, X1.1, M48, M102, N60, N82, AA, N61, and N83). This study shows that type BrIII genotype, circulating in Brazil in different hosts, can cause acute disease in a naturally infected animal host. The described case also involves the first reported occurrence of the 291 allele for the typing marker TUB2 in a type BrIII strain, emphasizing the genetic diversity of T. gondii in Brazil.
Asunto(s)
Conejos/parasitología , Toxoplasma/clasificación , Toxoplasmosis Animal/parasitología , Enfermedad Aguda , Alelos , Animales , Brasil , Resultado Fatal , Variación Genética , Genotipo , Hígado/parasitología , Hígado/patología , Repeticiones de Microsatélite/genética , Reacción en Cadena de la Polimerasa/veterinaria , Polimorfismo de Longitud del Fragmento de Restricción , Bazo/parasitología , Bazo/patología , Toxoplasma/genética , Toxoplasma/aislamiento & purificación , Toxoplasmosis Animal/diagnóstico , Toxoplasmosis Animal/patologíaRESUMEN
Genotyping and virulence studies of Toxoplasma gondii are essential to investigate the pathogenesis of strains circulating worldwide. In this study, eight T. gondii isolates obtained from a congenitally infected newborn, a calf, two cats, three dogs, and a wallaby from five states of México were genotyped by Mn-PCR-RFLP with 11 typing markers (SAG1, SAG2 5'3', alt. SAG2, SAG3, BTUB, GRA6, c22-8, c29-2, L358, PK1 and Apico), five virulence markers (CS3, ROP16, ROP17, ROP18 and ROP5), 15 microsatellite markers (TUB-2, W35, TgM-A, B18, B17, M33, IV.1, XI.1, M48, M102, N60, N82, AA, N61, N83), and sequencing. A phylogenetic network was built to determine the relationship between Mexican isolates and those reported worldwide. Six different genotypes were identified by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP), ToxoDB #8, #10, #28 (n = 3), #48, #116, and #282. Genotyping by microsatellite analysis differentiated the three PCR-RFLP genotype #28 isolates into two strains, revealing a total of seven microsatellite genotypes. Three different allele combinations of ROP18/ROP5 virulence markers were also found, 3/3, 1/1, and 4/1. The last two combinations are predicted to be highly virulent in the murine model. According to the phylogenetic network, the T. gondii strains studied here are related to archetypal strains I and III, but none are related to the strains previously reported in México. The genotypes identified in this study in different species of animals demonstrate the great genetic diversity of T. gondii in México. The ToxoDB-PCR-RFLP #28 genotype was found in three isolates from different hosts and states. Additionally, four of the isolates are predicted to be highly virulent in mice. The next step will be to perform in vitro and in vivo assays to determine the phenotype of these T. gondii isolates in murine models.
Asunto(s)
Toxoplasma , Toxoplasmosis Animal , Animales , Ratones , Perros , Genotipo , Filogenia , México , Polimorfismo de Longitud del Fragmento de Restricción , Variación GenéticaRESUMEN
In the present study, 51 strains of Toxoplasma gondii (T. gondii) were isolated from free-range chickens in the state of Mato Grosso, Midwestern Brazil, upon conducting bioassays in mice, and genotyped them using PCR-restriction fragment length polymorphism (PCR-RFLP) and 11 markers, including SAG1, SAG2 (5'3'SAG2 and alt. SAG2), SAG3, BTUB, GRA6, c22-8, c29-2, L358, PK1, Apico, and CS3. Fifty isolates were completely genotyped revealing 17 genotypes of T. gondii as follows: 12 matched using ToxoDB PCR-RFLP with the previously reported genotypes, including #6 type BrI (n = 4), #8 type BrIII (n = 7), #11 type BrII (n = 3), #14 (n = 1), #19 (n = 1), #41 (n = 1), #99 (n = 1), #109 (n = 4), #116 (n = 1), #140 (n = 2), #166 (n = 9), #190 (n = 1); and five genotypes have not been described before [#313 (n = 6), #314 (n = 1), #315 (n = 1), #316 (n = 1), #317 (n = 1)]. Moreover, mixed infections were identified in five isolates (TgCkBrMT8, TgCkBrMT9, TgCkBrMT33, TgCkBrMT38, and TgCkBrMT41). Additionally, genotype #190 was reported for the first time in chickens from Brazil. Our results corroborate with previous studies on T. gondii isolates identified in chickens from Brazil, thereby confirming their diversity, a typicality, and possibility of co-infection due to diï¬erent T. gondii strains present in the country.
Asunto(s)
Enfermedades de los Roedores , Toxoplasma , Toxoplasmosis Animal , Animales , Brasil/epidemiología , Pollos , Variación Genética , Genotipo , Ratones , Toxoplasma/genética , Toxoplasmosis Animal/epidemiologíaRESUMEN
Toxoplasma gondii infections are usually asymptomatic in pigs, and an acute clinical disease is rare in this host. This study aimed to determine the pathological and molecular aspects of an outbreak of fatal systemic toxoplasmosis in finishing pigs in Brazil. The outbreak occurred on a commercial finishing pig farm in the state of Santa Catarina in southern Brazil. The farm had 1500 pigs and 3.8% of mortality rate during the outbreak. The pigs had fever, anorexia, apathy, and locomotor deficits. Seven pigs were necropsied. Gross findings included multifocal to coalescent pale areas in skeletal muscles, lymphadenomegaly, hepatosplenomegaly, and non-colapsed lungs. The histological findings included granulomatous lymphadenitis, hepatitis and splenitis, necrotizing myositis, and lymphoplasmacytic interstitial pneumonia. Lung and liver lesions were occasionally accompanied by T. gondii parasitic structures. Positive immunolabeling for T. gondii tachyzoites and encysted bradyzoites was detected in all examined pigs. PCR-RFLP (11 markers) and microsatellite analysis (15 markers) identified the non-archetypal genotype #278 in pigs. This is the first report of systemic toxoplasmosis in pigs with muscle lesions and additionally shows the diversity of disease-causing T. gondii genotypes circulating in animals in Brazil.
Asunto(s)
Enfermedades de los Porcinos , Toxoplasma , Toxoplasmosis Animal , Porcinos , Animales , Toxoplasma/genética , Toxoplasmosis Animal/diagnóstico , Toxoplasmosis Animal/epidemiología , Toxoplasmosis Animal/parasitología , Polimorfismo de Longitud del Fragmento de Restricción , Genotipo , Brasil/epidemiología , Brotes de Enfermedades , Enfermedades de los Porcinos/epidemiología , Enfermedades de los Porcinos/parasitologíaRESUMEN
Studies on infectious and emerging diseases caused by bats have been increasing worldwide due to their well-recognised status as a reservoir species for various infectious agents as well as their close relationship to humans and animals. This study reports the molecular frequency and diversity of the parasites belonging to the Sarcocystidae family in bats in São Paulo state, Brazil. A total of 2892 tissue samples (brain and pectoral muscle/heart homogenates) from 1921 bats belonging to 36 species were collected, and the Sarcocystidae protozoan 18S ribosomal RNA encoding genes (18S rDNA) were detected by nested PCR and Sanger sequencing. The relative prevalence of Sarcocystidae species was 4.7% (91/1921) among 16 bat species, including insectivorous (n = 65), frugivorous (n = 13) and nectarivorous (n = 11) bats. From 66 sequenced positive samples, 50 were found to be suitable for analysis. Ten samples from insectivorous and nectarivorous bats showed 100% similarity with Neospora caninum (n = 1), Hammondia hammondi (n = 1), Cystoisospora canis (n = 1), Nephroisospora eptesici (n = 1), Sarcocystis (Frenkelia) glareoli (n = 1), and Toxoplasma gondii (n = 5). The 45 non-T. gondii samples revealed 15 different 18S rDNA alleles with identities varying from 96.1 to 100% with several Sarcocystidae species, which might suggest that bats can harbour a large variety of Sarcocystidae organisms. From the five T. gondii-positive tissue samples, three samples from two different bat specimens of the insectivorous Eumops glacinus were characterised using 11 PCR-restriction fragment length polymorphism (RFLP) markers, revealing the non-archetypal ToxoDB genotypes #6 (type BrI), which is one of the most prevalent in different hosts and regions from Brazil, and #69. We recommend the inclusion of T. gondii as a differential diagnosis for rabies and other neurological syndromes in bats.
RESUMEN
Aimed with this study to evaluate vertical transmission of Neospora caninum in naturally infected sheep and to monitor the kinetics of antibodies against this protozoon in their lambs. Therefore, 48 pregnant ewes, from five herds, were divided into two groups: G1 - positive for anti-N. caninum antibodies, with 19 animals; and G2 - seronegative, with 29 animals. Blood samples were taken from the ewes and their lambs, immediately after birth, before ingesting colostrum, and 2, 7, 14, 21, 28, 35, 42, 49 and 56 days after birth. Analysis on serum antibodies was performed using the indirect immunofluorescent antibody test. Among the 19 seropositive mothers, six (31.6%) gave birth to lambs seropositive before ingesting colostrum and it was found that these lambs remained positive until the end of the study (56 days). Only one of the lambs, from a ewe that presented an antibody titer of 200, seroconverted after ingestion of colostrum. All the lambs that had been born from negative mothers remained negative throughout the experimental period. It was concluded that transplacental transmission was an important form of diffusion of N. caninum in the herds studied and that seropositive lambs maintained circulating antibodies during the period analyzed.
Asunto(s)
Coccidiosis , Neospora , Enfermedades de las Ovejas , Animales , Anticuerpos Antiprotozoarios , Brasil , Coccidiosis/veterinaria , Femenino , Transmisión Vertical de Enfermedad Infecciosa/veterinaria , Cinética , Embarazo , OvinosRESUMEN
Through international trades, Europe, Africa and South America share a long history of exchanges, potentially of pathogens. We used the worldwide parasite Toxoplasma gondii to test the hypothesis of a historical influence on pathogen genetic diversity in Benin, a West African country with a longstanding sea trade history. In Africa, T. gondii spatial structure is still non-uniformly studied and very few articles have reported strain genetic diversity in fauna and clinical forms of human toxoplasmosis so far, even in African diaspora. Sera from 758 domestic animals (mainly poultry) in two coastal areas (Cotonou and Ouidah) and two inland areas (Parakou and Natitingou) were tested for T. gondii antibodies using a Modified Agglutination Test (MAT). The hearts and brains of 69 seropositive animals were collected for parasite isolation in a mouse bioassay. Forty-five strains were obtained and 39 genotypes could be described via 15-microsatellite genotyping, with a predominance of the autochthonous African lineage Africa 1 (36/39). The remaining genotypes were Africa 4 variant TUB2 (1/39) and two identical isolates (clone) of Type III (2/39). No difference in terms of genotype distribution between inland and coastal sampling sites was found. In particular, contrarily to what has been described in Senegal, no type II (mostly present in Europe) was isolated in poultry from coastal cities. This result seems to refute a possible role of European maritime trade in Benin despite it was one of the most important hubs during the slave trade period. However, the presence of the Africa 1 genotype in Brazil, predominant in Benin, and genetic analyses suggest that the triangular trade was a route for the intercontinental dissemination of genetic strains from Africa to South America. This supports the possibility of contamination in humans and animals with potentially imported virulent strains.
Asunto(s)
Comercio , Variación Genética , Enfermedades de las Aves de Corral/transmisión , Toxoplasma/genética , Toxoplasmosis Animal/parasitología , Toxoplasmosis Animal/transmisión , África Occidental/epidemiología , Animales , Benin , Pollos/parasitología , Europa (Continente)/epidemiología , Genética de Población , Genotipo , Geografía , Humanos , Ratones/parasitología , Repeticiones de Microsatélite , Filogenia , Filogeografía , Polimorfismo de Longitud del Fragmento de Restricción , Enfermedades de las Aves de Corral/parasitologíaRESUMEN
We aimed to describe the genetic diversity of Toxoplasma gondii strains isolated from domestic animals, wildlife and humans in the Midwestern Brazil. For this purpose, fragments of tissue samples (heart, brain and lung) from 35 dogs, four cats, 105 wildlife, and amniotic fluids from eight pregnant women were collected and submitted to mouse bioassay test. In a total, 22 isolates from nine dogs, one cat, ten wild animals and two women were obtained. The DNA was extracted from T. gondii isolates (lungs and brains of infected mice) and from "primary samples" (aliquots of tissue homogenate from wild animals and amniotic fluids from pregnant women) in order to screen using a Polymerase Chain Reaction (PCR) targeting a repeated 529-base pairs fragment of the T. gondii genome. All positive PCR samples were genotyped using restriction fragment length polymorphism (RFLP) analysis. To the best of our knowledge, this was the first study to report isolates of T. gondii from Leopardus pardalis, Crax fasciolata, and Dasyprocta azarae. Moreover, multilocus PCR-RFLP revealed 11 T. gondii RFLP genotypes, comprising nine previously described, including the archetypal lineage #2 type III (nâ¯=â¯1); two clonal Brazilian lineages, #6 type BrI (nâ¯=â¯1) and #8 type BrIII (nâ¯=â¯5); #14 (nâ¯=â¯2), #41 (nâ¯=â¯1), #108 (nâ¯=â¯1), #140 (nâ¯=â¯2), #166 (nâ¯=â¯4), #190 (nâ¯=â¯1), one potentially mixed, and two new described genotypes in two isolates. Our results confirmed the high diversity of T. gondii strains in Brazil, including identical genotypes circulating among humans, domestic dogs and wildlife.
Asunto(s)
Toxoplasma/clasificación , Toxoplasma/aislamiento & purificación , Animales , Animales Domésticos/parasitología , Animales Salvajes/parasitología , Brasil , Femenino , Variación Genética , Genotipo , Humanos , Ratones , Embarazo , Toxoplasma/genéticaRESUMEN
Dogs are hosts of the protozoans Toxoplasma gondii, which causes an important public health disease, and Neospora caninum. Studies that have evaluated toxoplasmosis and neosporosis for prolonged periods in dog populations are rare. We analyzed infection by both parasites in a domestic dog population over three consecutive years in São Paulo state, Brazil. In the 1st, 2nd and 3rd years of collection, 181, 193 and 172 domiciles were visited, and blood samples of 331, 371 and 348 dogs were collected for antibody serology, respectively. The seroprevalence of T. gondii in each year was 27.2%, 22.5% and 43.9%, respectively, and that of N. caninum was 7.8%, 4.8% and 6.8%, respectively. The incidence rates for T. gondii in the 2nd and 3rd collections were 13.2% and 30.0%, and those for N. caninum were 3.3% and 4.4%, respectively. Positive and negative serological conversions for both agents occurred at high frequencies during the study period. This study reveals the canine population's serological profile and demonstrates the constant exposure of dogs to the investigated pathogens, indicating the need for prevention and control measures in the region.
Asunto(s)
Coccidiosis , Enfermedades de los Perros , Neospora , Toxoplasma , Toxoplasmosis Animal , Animales , Anticuerpos Antiprotozoarios/sangre , Brasil/epidemiología , Coccidiosis/sangre , Coccidiosis/epidemiología , Coccidiosis/parasitología , Coccidiosis/veterinaria , Enfermedades de los Perros/epidemiología , Perros , Incidencia , Población Rural , Estudios Seroepidemiológicos , Toxoplasmosis Animal/sangre , Toxoplasmosis Animal/epidemiologíaRESUMEN
The accidental ingestion of treated recreational water is an important transmission route of waterborne protozoa worldwide. The present study aimed to provide the first evaluation of swimming pools in Brazil, analysing the presence of pathogenic protozoa (Toxoplasma gondii, Cryptosporidium spp. and Giardia spp.) by parasitological and molecular methods. A total of 57 samples were collected from 21 public swimming pools, either directly from the pool or filter backwash water and concentrated using the membrane filtration technique. Giardia cysts and Cryptosporidium oocysts were visualized by direct immunofluorescence assay after purification by immunomagnetic separation. Toxoplasma gondii oocysts were detected by autofluorescence visualization using the supernatant discarded during the purification step as a sample. Positive samples were submitted to molecular analysis. The molecular markers were used: SSU-rRNA, tpi, gdh and bg, for Giardia DNA amplification, and 18S rRNA gene fragment amplification was used for the Cryptosporidium oocysts. The 529-bp repeat element (REP529) fragment and the 35-fold repetitive B1 gene were employed as a target for T. gondii. Amplified products were submitted to sequencing and phylogenetic analysis. Giardia cysts were detected in 19.0% and Cryptosporidium oocysts in 9.5% of swimming pools. In one swimming pool (4.7%), both protozoa were detected on at least one occasion. Structures similar to T. gondii oocysts were detected in 33.3% of the samples, ranging from one to 23 per slide. Giardia was confirmed by DNA amplification in three swimming pools; Giardia duodenalis Assemblage A was identified by the phylogenetic positioning of the ß-giardin gene. Toxoplasma gondii DNA was detected in 14.2% of swimming pools. The present study represents the first report of the occurrence of T. gondii oocysts in swimming pools. Recreational activity in swimming pools contaminated by chlorine-resistant protozoa can represent a high risk of infection for bathers and swimmers.
Asunto(s)
Cryptosporidium/aislamiento & purificación , Giardia/aislamiento & purificación , Oocistos/aislamiento & purificación , Piscinas , Toxoplasma/aislamiento & purificación , Animales , Brasil , Técnica del Anticuerpo Fluorescente , Humanos , Factores de Riesgo , Agua/parasitologíaRESUMEN
The protozoan parasite Toxoplasma gondii can infect virtually all warm-blooded animals worldwide but little is known of its infection in the endangered giant anteaters (Myrmecophaga tridactyla). The present study found antibodies to T. gondii in 13 of 23 free-living M. tridactyla from the northwest region of São Paulo state, Brazil, by the Modified Agglutination Test (MAT, cut-off titer 1:25). Unfrozen tissues of seven M. tridactyla were bioassayed in mice and viable T. gondii (strain designated TgMytrBrSP1) isolated from one seropositive giant anteater. To our knowledge, this is a new host record for T. gondii. Genotyping using PCR-RFLP revealed the Brazilian clonal Type BrIII genotype, and a unique non-archetypal genotype was revealed by microsatellite analysis.
Asunto(s)
Euterios/parasitología , Toxoplasma/genética , Toxoplasmosis Animal/parasitología , Animales , Genotipo , Técnicas de Genotipaje , RatonesRESUMEN
Genotyping of Toxoplasma gondii remains a relevant topic of study, since genotypes can be related to the presentation and severity of toxoplasmosis. To date, 292 restriction fragment length polymorphism genotypes have been described around the world. Serosurveys in southeastern Mexico have documented exposure in over 70% of people and certain animals. Recently, we have described new genotypes and mixed infections in feral cats from Quintana Roo. Thus, the aim of this study was to genotype T. gondii and to describe its genetic variability, from naturally infected stray dogs of Chiapas, which has different geographical and climatic conditions from those found at the Yucatan Peninsula and the other parts of the country. Eleven stray dogs were captured and bled to obtain DNA, and then they were euthanized to perform necropsies and to collect target tissues. Diagnosis of T. gondii was done by quantitative real-time PCR (qPCR) and endpoint PCR. Genotyping was carried out, amplifying 12 polymorphic markers and 15 microsatellites. Atypical SAG3 gene products were cloned and sequenced. All blood samples of dogs were positive to T. gondii DNA by PCR. Two isolates were obtained from pooled heart and diaphragm tissue of two dogs. Two complete PCR-RFLP genotypes were identified (type BrIII and #28). Four animals had mixed infections. A new RFLP atypical allele for the SAG3 marker was observed; cloning and sequencing analysis of this locus revealed mixed infection by a strain identical to GT1, and one type I × II intragenic recombinant. The microsatellite analysis revealed that both isolates are atypical. Thus, atypical new genotypes of T. gondii and mixed infections were found in dogs of Chiapas. The results found here and in genotyping studies in México suggest that the southeastern region favours wide genetic diversity of T. gondii and the possible presence of virulent genotypes such as those found in central and South America.
Asunto(s)
Glicoproteínas de Membrana/genética , Proteínas Protozoarias/genética , Toxoplasma/genética , Toxoplasmosis Animal , Animales , Sangre/parasitología , ADN Protozoario/genética , Perros , Marcadores Genéticos , Variación Genética , Genotipo , Humanos , México/epidemiología , Repeticiones de Microsatélite/genética , Filogenia , Polimorfismo de Longitud del Fragmento de Restricción/genética , América del Sur , Toxoplasma/clasificación , Toxoplasma/aislamiento & purificación , Toxoplasmosis/epidemiología , Toxoplasmosis Animal/epidemiología , ZoonosisRESUMEN
Neospora caninum is a heteroxenous protozoa, whose definitive hosts are canids and intermediate hosts are herbivores, and is of great importance in cattle. The objectives of this study were to determine the prevalence of N. caninum in dairy cattle and dogs, to detect the presence of the protozoa at the molecular level in aborted fetuses, and to identify the risk factors associated with infection in properties in the western region of the state of Paraná. For this study, 600 bovine serum samples from 60 properties, 163 canine serum samples from 52 properties and 17 bovine fetuses from nine properties were collected. Data were collected using an epidemiological questionnaire to verify the risk factors. Serum samples were analyzed using the indirect fluorescent antibody test. Fetal tissues were analyzed using polymerase chain reaction and subsequent DNA sequencing. Of the bovine samples, 23.67% were positive for N. caninum. Among the canine samples, 11.66% were positive for N. caninum. Risk factors in cattle were history of abortion, low milk production, extensive breeding, and Jersey breed (p<0.05). Protozoan DNA was detected in 52.94% of the 17 fetuses and the sequencing presented high similarity with N. caninum.