RESUMO
The aim of this longitudinal study was to characterize the parasitemia of Neospora caninum and the associated immunological parameters in naturally infected beef cows for 10 months. The following groups were established: Neospora caninum seropositive pregnant cows (+Preg, n = 7), seropositive non-pregnant cows (+Npreg, n = 7), seronegative pregnant cows (-Preg, n = 4), and seronegative non-pregnant cows (-Npreg, n = 4). Several samples were obtained for absolute and relative leukocyte counting, cytokines IL-10, IL-12, α-TNF, and γ-IFN quantification, specific IgG, IgG1, and IgG2 and avidity and N. caninum DNA molecular detection and quantification. The +Preg group had a higher frequency and concentration of N. caninum DNA in PBMC in the last third of pregnancy compared to +Npreg (p <0.05), with 22 and 8% of detection, respectively. Parasitemia correlated positively with IgG titers and negatively with IgG1/IgG2 ratio (p <0.05). On day 222 of the assay, the +Preg group had the lowest total leukocyte counting (p <0.05). The +Preg group had a higher concentration of IgG and higher avidity in the last third of gestation compared to +Npreg (p <0.05). Avidity correlated with total IgG and IgG2 (p <0.05). All +Preg cows gave birth to clinically healthy but seropositive calves before colostrum intake, therefore, the congenital transmission was 100% efficient. Only a complete N. caninum genotype from a placenta and a partial genotype from cow #3 of the group +Preg were achieved by multilocus microsatellite analysis. Overall, N. caninum parasitemia is frequent in seropositive beef cows during the last third of gestation. This correlates with higher antibody levels and a decrease in total leukocyte counting. The precise timing of the parasitemia may be used for diagnosis purposes and/or for design strategies to avoid vertical transmission. Further studies are needed to identify the immune molecular mechanisms that favor parasitemia during gestation in chronically infected cattle.
RESUMO
Dogs play a potential role as reservoirs for zoonotic parasites, being especially problematic uncontrolled dog populations such as stray and farm dogs with access to populated areas. In order to investigate the prevalence of canine intestinal parasites in at-risk dog populations, we tested a total of 233 faecal samples shed by stray and dairy farm dogs from northern Spain. Telemann method was used to detect the presence of eggs and (oo)cysts of common dog intestinal parasites and Cryptosporidium was detected by PCR. One hundred and forty eight out of 233 samples (63.5%) were positive for at least one intestinal parasite, being Ancylostomidae (35.6%; 83/233) and Trichuris (35.2%; 82/233) the parasites most frequently identified. Cryptosporidium DNA was not detected in any of the faecal samples analysed. The overall prevalence was significantly higher in stray dogs than in farm dogs (72.5% vs 58.8%). Specifically, stray dogs had a significantly higher prevalence of Ancylostomatidae, Toxocara, Toxascaris and Taenidae. These dog populations are an important source of environmental contamination with intestinal parasite forms, which could be of significance to animal and human health.
Assuntos
Doenças do Cão , Fazendas , Enteropatias Parasitárias/veterinária , Animais , Criptosporidiose/epidemiologia , Cryptosporidium , Doenças do Cão/epidemiologia , Doenças do Cão/parasitologia , Cães , Fazendas/estatística & dados numéricos , Fezes/parasitologia , Humanos , Enteropatias Parasitárias/epidemiologia , Enteropatias Parasitárias/parasitologia , Prevalência , Espanha/epidemiologiaRESUMO
Abstract Dogs play a potential role as reservoirs for zoonotic parasites, being especially problematic uncontrolled dog populations such as stray and farm dogs with access to populated areas. In order to investigate the prevalence of canine intestinal parasites in at-risk dog populations, we tested a total of 233 faecal samples shed by stray and dairy farm dogs from northern Spain. Telemann method was used to detect the presence of eggs and (oo)cysts of common dog intestinal parasites and Cryptosporidium was detected by PCR. One hundred and forty eight out of 233 samples (63.5%) were positive for at least one intestinal parasite, being Ancylostomidae (35.6%; 83/233) and Trichuris (35.2%; 82/233) the parasites most frequently identified. Cryptosporidium DNA was not detected in any of the faecal samples analysed. The overall prevalence was significantly higher in stray dogs than in farm dogs (72.5% vs 58.8%). Specifically, stray dogs had a significantly higher prevalence of Ancylostomatidae, Toxocara, Toxascaris and Taenidae. These dog populations are an important source of environmental contamination with intestinal parasite forms, which could be of significance to animal and human health.
Resumo Os cães desempenham um importante papel como reservatório de parasitos zoonóticos, sendo especialmente problemáticas as populações descontroladas, como a de cães errantes e de fazenda, com acesso às áreas povoadas. Para investigar a prevalência de parasitos intestinais em populações caninas de risco, foram analisadas 233 amostras fecais provenientes de cães de fazendas leiteiras e errantes do norte da Espanha. O método Telemann foi utilizado para detectar ovos, cistos e oocistos dos parasitos caninos mais comuns e para a detecção de Cryptosporidium foi utilizada a técnica da PCR. Cento e quarenta e oito de 233 amostras analisadas (63,5%) foram positivas para pelo menos um parasito intestinal, sendo Ancyostomatidae (35,6%; 83/233) e Trichuris sp. (35,2%; 82/233) os parasitos identificados com maior frequência. O DNA de Cryptosporidium sp. não foi detectado em nenhuma das amostras fecais analisadas. A prevalência geral foi significativamente maior em cães errantes do que em cães de fazenda (72,5% vs 58,8%). Especificamente, os cães errantes tiveram prevalência maior para Ancylostomatidae, Toxocara, Toxascaris e Taenidae. Essas populações de cães são importantes fontes de contaminação ambiental, pois eliminam formas de vida desses parasitos, que podem ter impacto na saúde animal e humana.
Assuntos
Animais , Cães , Doenças do Cão/parasitologia , Doenças do Cão/epidemiologia , Fazendas/estatística & dados numéricos , Enteropatias Parasitárias/veterinária , Espanha/epidemiologia , Prevalência , Criptosporidiose/epidemiologia , Cryptosporidium , Fezes/parasitologia , Enteropatias Parasitárias/parasitologia , Enteropatias Parasitárias/epidemiologiaRESUMO
Dogs play a potential role as reservoirs for zoonotic parasites, being especially problematic uncontrolled dog populations such as stray and farm dogs with access to populated areas. In order to investigate the prevalence of canine intestinal parasites in at-risk dog populations, we tested a total of 233 faecal samples shed by stray and dairy farm dogs from northern Spain. Telemann method was used to detect the presence of eggs and (oo)cysts of common dog intestinal parasites and Cryptosporidium was detected by PCR. One hundred and forty eight out of 233 samples (63.5%) were positive for at least one intestinal parasite, being Ancylostomidae (35.6%; 83/233) and Trichuris (35.2%; 82/233) the parasites most frequently identified. Cryptosporidium DNA was not detected in any of the faecal samples analysed. The overall prevalence was significantly higher in stray dogs than in farm dogs (72.5% vs 58.8%). Specifically, stray dogs had a significantly higher prevalence of Ancylostomatidae, Toxocara, Toxascaris and Taenidae. These dog populations are an important source of environmental contamination with intestinal parasite forms, which could be of significance to animal and human health.(AU)
Os cães desempenham um importante papel como reservatório de parasitos zoonóticos, sendo especialmente problemáticas as populações descontroladas, como a de cães errantes e de fazenda, com acesso às áreas povoadas. Para investigar a prevalência de parasitos intestinais em populações caninas de risco, foram analisadas 233 amostras fecais provenientes de cães de fazendas leiteiras e errantes do norte da Espanha. O método Telemann foi utilizado para detectar ovos, cistos e oocistos dos parasitos caninos mais comuns e para a detecção de Cryptosporidium foi utilizada a técnica da PCR. Cento e quarenta e oito de 233 amostras analisadas (63,5%) foram positivas para pelo menos um parasito intestinal, sendo Ancyostomatidae (35,6%; 83/233) e Trichuris sp. (35,2%; 82/233) os parasitos identificados com maior frequência. O DNA de Cryptosporidium sp. não foi detectado em nenhuma das amostras fecais analisadas. A prevalência geral foi significativamente maior em cães errantes do que em cães de fazenda (72,5% vs 58,8%). Especificamente, os cães errantes tiveram prevalência maior para Ancylostomatidae, Toxocara, Toxascaris e Taenidae. Essas populações de cães são importantes fontes de contaminação ambiental, pois eliminam formas de vida desses parasitos, que podem ter impacto na saúde animal e humana.(AU)
Assuntos
Animais , Cães , Cães/parasitologia , Intestinos/parasitologia , Criptosporidiose/diagnóstico , Ancylostoma , TrichurisRESUMO
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 18â¯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.
Assuntos
Coccidiose/veterinária , Feto/parasitologia , Genótipo , Doenças das Cabras/diagnóstico , Repetições de Microssatélites , Neospora/genética , Animais , Argentina , Coccidiose/diagnóstico , Coccidiose/parasitologia , Feminino , Doenças das Cabras/parasitologia , Cabras , Tipagem de Sequências Multilocus/veterinária , Gravidez , Complicações Parasitárias na Gravidez/diagnóstico , Complicações Parasitárias na Gravidez/parasitologia , Complicações Parasitárias na Gravidez/veterináriaRESUMO
Coxiella burnetii, an intracellular bacterium, is the agent of Q fever/coxiellosis, a worldwide zoonosis. Dairy animals are the primary reservoirs of C. burnetii, and although the disease is usually asymptomatic or subclinical, abortion is a serious clinical outcome among small ruminants. This study was conducted to investigate C. burnetii seroprevalence and infection In a flock of dairy goats in Brazil. Serum samples from 312 goats collected from a dairy goat flock with a history of reproductive failure were tested by a commercial ELISA (LSIVet Ruminant Q Fever - Serum/Milk; Thermo Fisher Scientific, Lissieu, France) for anti-C. burnetii IgG antibodies. Samples of cotyledons from 23 placentas were analyzed by nested PCR for the presence of the bacterial DNA. ELISA seroreactivity was found in 55.1% (172/312; 95% CIâ¯=â¯49.4%-60.7%) of the serum samples analyzed. C. burnetii DNA was detected in 8.7% (2/23) of the placental samples tested, where both animals were also seropositive. This study reports the first description of C. burnetii infection in an abortion outbreak in goats in Brazil. The results point out to the importance of including this disease in animal and public health surveillance programs as well as into the list of abortive diseases in goats in Brazil.
Assuntos
Coxiella burnetii/isolamento & purificação , Indústria de Laticínios , Doenças das Cabras/microbiologia , Cabras/microbiologia , Febre Q/veterinária , Animais , Brasil/epidemiologia , Cruzamento , Coxiella burnetii/genética , DNA Bacteriano/análise , Surtos de Doenças/veterinária , Ensaio de Imunoadsorção Enzimática/veterinária , Feminino , Doenças das Cabras/epidemiologia , Leite/microbiologia , Placenta/microbiologia , Reação em Cadeia da Polimerase/veterinária , Gravidez , Febre Q/epidemiologia , Febre Q/microbiologia , Reprodução , Estudos Soroepidemiológicos , Zoonoses/epidemiologiaRESUMO
The purpose of this study was to perform genotypic characterization and to evaluate the virulence of Toxoplasma gondii obtained from aborted fetuses in an abortion outbreak in goats from northeastern Brazil. Brain samples from 32 fetuses were submitted to mouse bioassay for T. gondii isolation. Two isolates were obtained and subjected to genotypic characterization. Isolate virulence was evaluated using murine model in different doses (from 105 to 101 tachyzoites/mL). In genotyping, both isolates were classified as clonal lineage type II (genotype #1 ToxoDB) and showed to be virulent for mice. This is the first description of genotype #1 in cases of goat abortion, showing the circulation of virulent T. gondii isolate producing reproductive disorders in pregnant goat.
Assuntos
Aborto Animal/parasitologia , Surtos de Doenças/veterinária , Doenças das Cabras/parasitologia , Toxoplasma/classificação , Toxoplasmose Animal/parasitologia , Aborto Animal/epidemiologia , Animais , Anticorpos Antiprotozoários/sangue , Bioensaio/veterinária , Encéfalo/embriologia , Encéfalo/parasitologia , Brasil/epidemiologia , Linhagem Celular , Chlorocebus aethiops , Genótipo , Técnicas de Genotipagem/veterinária , Doenças das Cabras/epidemiologia , Cabras , Camundongos , Filogenia , Reação em Cadeia da Polimerase/veterinária , Polimorfismo de Fragmento de Restrição , Toxoplasma/genética , Toxoplasma/imunologia , Toxoplasma/patogenicidade , Toxoplasmose Animal/complicações , Toxoplasmose Animal/epidemiologia , VirulênciaRESUMO
Bovine neosporosis has become a disease of international concern as it is among the main causes of abortion in cattle. Viable N. caninum has been isolated from brains of fetuses and neonatal calves, and there is no report of isolation of tachyzoites from kidney. Also, detailed information about the genetic diversity of N. caninum is scarce. N. caninum tachyzoites were isolated from the kidney and the brain of an aborted 4-month-old bovine foetus. The parasite was confirmed to be N. caninum by PCR. The tachyzoites of the new isolate, named BNC-PR4, were propagated in Vero cell cultures. Pathogenicity of the parasite was examined in BALB/c mice. Mice inoculated intraperitoneally with BNC-PR4 failed to yield clinical signs of disease and did not induce severe brain lesions, suggesting a bovine isolate with low virulence. The N. caninum-positive DNA sample was further analyzed by multilocus microsatellite (MS) genotyping for MS4, MS5, MS6A, MS6B, MS7, MS8, MS10, MS12, and MS21. Multilocus-microsatellite genotyping revealed a unique genetic profile that differed from previously reported isolates.
Assuntos
Feto Abortado/parasitologia , Encéfalo/parasitologia , Doenças dos Bovinos/parasitologia , Coccidiose/veterinária , Rim/parasitologia , Neospora/isolamento & purificação , Aborto Animal/parasitologia , Animais , Encéfalo/embriologia , Brasil , Bovinos , Doenças dos Bovinos/embriologia , Chlorocebus aethiops , Coccidiose/embriologia , Coccidiose/parasitologia , DNA de Protozoário/química , DNA de Protozoário/isolamento & purificação , Indústria de Laticínios , Feminino , Rim/embriologia , Camundongos , Camundongos Endogâmicos BALB C , Neospora/genética , Neospora/patogenicidade , Células Vero , VirulênciaRESUMO
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.
Assuntos
Doenças dos Bovinos/parasitologia , Coccidiose/veterinária , Neospora/isolamento & purificação , Animais , Anticorpos Antiprotozoários/sangue , Bioensaio/veterinária , Encéfalo/parasitologia , Brasil , Bovinos , Coccidiose/parasitologia , DNA de Protozoário/química , Cães , Fezes/parasitologia , Feminino , Técnica Indireta de Fluorescência para Anticorpo/veterinária , Técnicas de Genotipagem/veterinária , Gerbillinae , Masculino , Repetições de Microssatélites , Neospora/genética , Neospora/imunologia , Oocistos/genética , Oocistos/imunologia , Oocistos/isolamento & purificação , Reação em Cadeia da Polimerase/veterinária , Polimorfismo de Fragmento de Restrição , Soro/parasitologiaRESUMO
The cyst-forming protozoan parasite Neospora caninum is one of the main causes of bovine abortion worldwide and is of great economic importance in the cattle industry. Recent studies have revealed extensive genetic variation among N. caninum isolates based on multilocus microsatellite genotyping. Currently, the most extensive study reported is based on the N. caninum genotyping of 96 samples from four countries on two continents (Spain, Argentina, Germany and Scotland) that demonstrate different clusters of multilocus genotypes (MLGs) implicated in cattle abortions as well as the population sub-structuring of N. caninum, which is partially associated with the geographical origin. The aim of this study was to genotype N. caninum from aborted bovine foetuses that originated from Mexico within the region of Aguascalientes and to investigate their genetic diversity. Parasite DNA was detected in 27 out of the 63 analysed foetuses recovered from 10 different herds. Complete or nearly complete profiles based on 9 microsatellite markers were obtained from 11 samples. Diverse N. caninum MLGs were implicated in the occurrence of abortion in each herd. All of the Mexican MLGs differed from the MLGs previously determined for the Argentinean, Spanish, German and Scottish N. caninum populations. The Mexican MLGs failed to cluster by eBURST analyses. The MLG relationships using PCoA showed a close genetic relationship between the Spanish population and a portion of the Mexican population, but a more distant genetic relationship with the Argentinean genotypes. These results demonstrate the genetic diversity of N. caninum in the studied areas that differed from other populations of N. caninum around the world.