RESUMO
Neospora caninum is recognised for causing cattle abortion, provoking severe economic losses in the livestock industry worldwide. The aim of the present study was to evaluate the reactivation and foetal infection in pregnant heifers inoculated with live N. caninum tachyzoites before puberty. A total of 15 30-month-old pregnant heifers were allocated into four groups: animals inoculated with live tachyzoites of NC-Argentina LP1 isolate before puberty and challenged with live tachyzoites of NC-1 strain at 210 days of gestation (DG) (Group A); animals mock inoculated before puberty and challenged with NC-1 strain at 210 DG (Group B), animals inoculated before puberty but not subsequently challenged (Group C); and noninfected and nonchallenged animals (Group D). The results of this study showed that 100% of animals infected before puberty (Groups A and C) suffered reactivation of the infection at the seventh month of gestation. In addition, in three and two calves from Groups A and C, respectively, congenital infection was confirmed. Interestingly, we provide evidence that the use of live N. caninum tachyzoites in young animals as a strategy to induce protection is neither safe nor effective.
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
Neospora caninum is a major reproductive disease in cattle worldwide. In the Argentinian Humid Pampa, the seroprevalence, incidence of abortions, and economic losses due to neosporosis are considerably higher in dairy than in beef cattle. Despite this, we recently demonstrated that N. caninum subpopulations are indistinctly distributed in both dairy and beef production systems. The association between genotypic characteristics defined by microsatellite analysis and the virulence of the different strains-particularly with regard to the severity and extension of histological lesions-is largely unknown. Herein, we used a morphometric approach to analyze encephalic lesions in 62 bovine fetuses spontaneously infected by N. caninum. Morphometric parameters (average size of focal lesions, number of foci/cm2 and the percentage of the section affected by lesions) were compared according to the N. caninum subpopulations found in our previous microsatellite genotyping analysis, animal biotype (beef versus dairy), and fetal age (second stage of gestation versus third stage). The average size of the lesions differed significantly among fetuses with different gestational ages; however, no significant differences among animal biotypes or genotypic patterns were found. Further research into the genetic, molecular, and husbandry factors that could account for this greater impact in Argentinian dairy herds is needed.
Assuntos
Aborto Animal , Doenças dos Bovinos , Coccidiose , Neospora , Animais , Anticorpos Antiprotozoários , Argentina , Bovinos , Doenças dos Bovinos/epidemiologia , Doenças dos Bovinos/parasitologia , Coccidiose/veterinária , Feminino , Feto/parasitologia , Feto/patologia , Neospora/genética , Neospora/imunologia , Gravidez , Estudos SoroepidemiológicosRESUMO
Neospora caninum is an apicomplexan protozoan and a major cause of abortion in cattle worldwide. In the Argentinian Humid Pampa, bovine neosporosis causes severe economic losses. Despite this, information on the genetic structure of N. caninum in this region is limited. Therefore, this study aimed to genetically characterize N. caninum isolates associated with bovine abortion in the Humid Pampa region. For this purpose, spontaneous bovine fetal tissues submitted for diagnosis to the Veterinary Diagnostic Service at INTA Balcarce during 2008-2019 were assessed by PCR, indirect fluorescent antibody test (IFAT), and histologic analysis. PCR-positive samples were tested by multilocus microsatellite genotyping (MLGs) using 9 microsatellite markers. Thirty-one different genotypes were identified from 32 samples with at least seven markers. Argentinian MLGs were grouped into two clonal clusters when analyzed using eBURST network and principal coordinate analysis. No segregation based on the year of collection, animal biotype, or geographic origin was observed. In addition, the presence of linkage disequilibrium supported the clonal propagation of Argentinian MLGs. One Argentinian subpopulation was associated with isolates from Spain, Uruguay, Brazil, and Mexico, and the other one was linked to isolates from Scotland, Spain, and Germany. These findings reveal the presence of two clonal subpopulations of N. caninum in the Humid Pampa.
Assuntos
Doenças dos Bovinos/parasitologia , Coccidiose/veterinária , Repetições de Microssatélites/genética , Neospora/genética , Animais , Argentina , Bovinos , Doenças dos Bovinos/genética , Feminino , Feto , Técnica Indireta de Fluorescência para Anticorpo , Variação Genética/genética , Genótipo , Desequilíbrio de Ligação/genética , Neospora/imunologia , Reação em Cadeia da Polimerase , GravidezRESUMO
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
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
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
The aim of the present study was to characterize the specific immune response in prepubertal female calves inoculated with Neospora caninum. Forty-eight N. caninum-seronegative 6-month-old Angus female calves were randomly allocated into two groups: group A calves were inoculated subcutaneously (sc) with 1 × 106 tachyzoites of the low virulence NC-Argentina LP1 isolate in sterile phosphate-buffered saline (PBS); group B calves were mock inoculated sc with sterile PBS. Calves from group A developed a specific immune response characterized by the production of IgG antibodies and the expression of IFN-γ and TNF-α cytokines. Animals did not present any febrile reaction or reactions at the site of inoculation. Although chronic N. caninum infection was developed in 50% of calves of group A after inoculation, according to the presence of antibodies against rNc-SAG4, antigen characteristic of bradyzoites, N. caninum antibodies dropped below the cut-off of ELISA from day 210 post-inoculation onwards. Future trials using the same group of inoculated animals will allow the characterization of the evolution of the immune response during pregnancy and to determine whether the immunization with the local isolate is able to prevent congenital transmission and to protect against heterologous challenges.
Assuntos
Anticorpos Antiprotozoários/sangue , Antígenos de Protozoários/imunologia , Doenças dos Bovinos/imunologia , Coccidiose/veterinária , Neospora/imunologia , Animais , Bovinos , Doenças dos Bovinos/parasitologia , Coccidiose/imunologia , Coccidiose/parasitologia , Citocinas/metabolismo , Feminino , Imunização/veterinária , Neospora/patogenicidade , Distribuição AleatóriaRESUMO
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
We carried out an inter-laboratory trial to compare the serological tests commonly used for the detection of specific Neospora caninum antibodies in cattle in Ibero-American countries. A total of eight laboratories participated from the following countries: Argentina (n = 4), Brazil (n = 1), Peru (n = 1), Mexico (n = 1), and Spain (n = 1). A blind panel of well-characterized cattle sera (n = 143) and sera representative of the target population (n = 351) was tested by seven in-house indirect fluorescent antibody tests (IFATs 1-7) and three enzyme-linked immunosorbent assays (ELISAs 1-3; two in-house and one commercial). Diagnostic performance of the serological tests was calculated and compared according to the following criteria: (1) the "Pre-test information," which uses previous epidemiological and serological data; (2) the "Majority of tests," which classifies a serum as positive or negative according to the results obtained by most tests evaluated. Unexpectedly, six tests showed either sensitivity (Se) or specificity (Sp) values lower than 90%. In contrast, the best tests in terms of Se, Sp, and area under the ROC curve (AUC) values were IFAT 1 and optimized ELISA 1 and ELISA 2. We evaluated a high number of IFATs, which are the most widely used tests in Ibero-America. The significant discordances observed among the tests regardless of the criteria employed hinder control programs and urge the use of a common test or with similar performances to either the optimized IFAT 1 and ELISAs 1 and 2.
Assuntos
Doenças dos Bovinos/diagnóstico , Coccidiose/veterinária , Ensaio de Imunoadsorção Enzimática/veterinária , Técnica Indireta de Fluorescência para Anticorpo/veterinária , Neospora/isolamento & purificação , Testes Sorológicos/veterinária , Animais , Anticorpos Antiprotozoários/análise , Argentina , Brasil , Bovinos , Coccidiose/diagnóstico , Ensaio de Imunoadsorção Enzimática/métodos , Técnica Indireta de Fluorescência para Anticorpo/métodos , México , Peru , Testes Sorológicos/métodos , EspanhaRESUMO
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
BACKGROUND: This work studied the natural infection of Neospora caninum during the first gestation of heifers in a dairy farm in animals consuming a ration co ntaminated naturally with Zearalenone (ZEA), and to find out effect of mycotoxin in the levels of estrogen (E) and progesterone (P4) and that's relation to the infection to N. caninum and in the abortions. METHODS: The study was conducted in a dairy farm located in El Llano municipality, in Aguascalientes, Mexico, in 2013. Two groups were formed, the group "A" with 20 seronegative animals to N. caninum, and group "B" with 20 seropositive. Once a month was determined the levels of total IgG to N. caninum, the serum concentration of E and P4, and the level of ZEA in the ration; in cases of abortion, fetal brain samples were taken to identify the presence of N. caninum DNA. RESULTS: In group "A", was observed two subgroups: seronegative (60%) and seroconverted (40%), and three abortions. In group "B", all animals maintain their serostatus, and three animals aborted. All abortions were positive for N. caninum DNA. The level of ZEA in the ration has an average of 426 µg/kg; during the gestation did not identify that animals suffer any alteration in the levels of E or P4. No statistical differences among the studied variables (levels of E and P) in time (nine months of gestation) were detected. It was not identified any interaction with the natural exposure to ZEA intake in any of the groups under study. CONCLUSION: The chronic ingestion of ZEA does not affect serum concentrations of E and P4 during gestation of heifers under study and cannot be related to the infection for N. caninum and the abortion.
RESUMO
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.
Assuntos
Aborto Animal/parasitologia , Doenças dos Bovinos/parasitologia , Coccidiose/veterinária , Variação Genética , Neospora/genética , Complicações Parasitárias na Gravidez/veterinária , Animais , Bovinos , Coccidiose/parasitologia , Feminino , Feto/parasitologia , Genótipo , México , Repetições de Microssatélites/genética , Neospora/isolamento & purificação , Gravidez , Complicações Parasitárias na Gravidez/parasitologiaRESUMO
Here, we assessed outcome of experimental infection by Neospora caninum in goats intravenously inoculated with 10(6) tachyzoites of the Nc-Spain7 isolate at 40 (G1), 90 (G2) and 120 (G3) days of gestation. Infected goats had fever between 5 and 9 days post inoculation (dpi); all were seropositive at the time of abortion/birth. Foetal death occurred in G1 from 10 to 21 dpi (n = 7) and in G2 from 27 to 35 dpi (n = 4). Goats in G2 also had seropositive stillbirth (n = 1) and healthy kids (n = 2). G3 goats (n = 7) had 3 seropositive and 3 seronegative weak kids, and 2 seronegative healthy kids. Parasite DNA detection in placentomes was 100% in G2, 85.7% in G3 and in G1 was detected only in placentomes from the goats with foetal losses from 17 dpi (100%). Parasites were detected in foetal/kid brain (>85.7%) and liver (≥ 50%) of G2 and G3, and in G1 after 17 dpi (100%). The highest parasite loads were detected in the placentomes of G1 from 17 dpi and G2, and in foetal tissues of G1 from 17 dpi and G3. Multifocal necrotic lesions were observed in the placentas of the three groups, but they were larger and more frequent in G1 and G2. Similar lesions were observed in foetal tissues, but they were more frequent in G3. These findings suggest that, as observed in cattle and sheep, the clinical consequences of N. caninum in pregnant goats are dependent in part on the time of gestation when animals were infected.
Assuntos
Coccidiose/veterinária , Doenças das Cabras/transmissão , Transmissão Vertical de Doenças Infecciosas/veterinária , Neospora/fisiologia , Animais , Coccidiose/parasitologia , Coccidiose/transmissão , Feminino , Idade Gestacional , Doenças das Cabras/parasitologia , Cabras , Carga Parasitária/veterinária , Placenta/parasitologia , GravidezRESUMO
The Neospora caninum microsatellite markers were applied to clinical samples. Genotyping technology involving fluorescently labelled DNA fragment analysis was used in combination with DNA sequencing for markers with complex repetitive sequences. Nineteen DNA samples from 15 brains and four hearts of naturally infected non-aborted zebuine foetuses from abattoirs in Goiás, Brazil. N. caninum had been detected in these foetuses by nested-PCR of the internal transcribed spacer-1 rRNA region, and the samples were analysed using these microsatellites. Seven complete or nearly complete allele profiles were obtained from six foetuses. Three distinct profiles of N. caninum were identified in a unique microregion (Meia Ponte) of Goiás. Two alleles for the same marker were detected in a unique foetus that was probably infected with two different strains. A new allele for one of the microsatellites is described. The multilocus analysis performed here revealed a preliminary means of discriminating between individual strains according to their geographical origins. These are the first results that have been obtained regarding the molecular characterisation of strains of N. caninum from infected zebuine foetuses in South America and reveal for the first time that there are genotypic differences in the strains that are responsible for foetal transmission in zebuine foetuses.
Assuntos
Doenças dos Bovinos/parasitologia , Coccidiose/veterinária , Transmissão Vertical de Doenças Infecciosas/veterinária , Neospora/genética , Matadouros , Animais , Brasil/epidemiologia , Bovinos , Doenças dos Bovinos/epidemiologia , Coccidiose/epidemiologia , Coccidiose/parasitologia , DNA de Protozoário/química , DNA de Protozoário/genética , Feminino , Feto/parasitologia , Genótipo , Geografia , Repetições de Microssatélites/genética , Tipagem de Sequências Multilocus/veterinária , Neospora/classificação , Neospora/isolamento & purificação , Reação em Cadeia da Polimerase/veterinária , Análise de Sequência de DNA/veterináriaRESUMO
The objective of this study was to investigate the presence of Toxoplasma gondii- and Neospora caninum-specific antibodies in domestic South American camelids (SAC) (llamas and alpacas) from the Peruvian Andes through a cross-sectional study. A wide panel of serum samples collected from 1,845 llamas and 2,874 alpacas from the two main SAC production areas of Peru was selected. Immunofluorescence antibody technique was employed to detect and titrate specific anti-T. gondii and anti-N. caninum immunoglobulins G in serum samples. The association between T. gondii and N. caninum seroprevalence and the geographical origin (Central and South Peruvian Andes) was evaluated. Anti-T. gondii antibodies were found in 460 (24.9 %) llamas and 706 (24.6 %) alpacas, whereas anti-N. caninum antibodies were detected in 153 (8.3 %) llamas and 425 (14.8 %) alpacas. Toxoplasma gondii infection was strongly associated with the South Peruvian Andes where moderate climate conditions, larger human population, compared to the Central region, and the presence of wildlife definitive hosts could favor horizontal transmission to SAC. In contrast, N. caninum infection was not associated with the geographical region. These results indicate that T. gondii and N. caninum infections are highly and moderately widespread, respectively, in both species of domestic SAC studied in the sampled areas and appropriate control measures should be undertaken to reduce the prevalence of both parasitic infections.
Assuntos
Animais Domésticos/parasitologia , Camelídeos Americanos/parasitologia , Coccidiose/epidemiologia , Coccidiose/veterinária , Toxoplasmose Animal/epidemiologia , Animais , Anticorpos Antiprotozoários/sangue , Estudos Transversais , Técnica Direta de Fluorescência para Anticorpo/veterinária , Geografia , Neospora/imunologia , Razão de Chances , Peru/epidemiologia , Densidade Demográfica , Estudos Soroepidemiológicos , Toxoplasma/imunologiaRESUMO
To determine the impact of farming over vicuña population in Peru, serum samples were collected from 207 vicuñas (126 captive vicuñas and 81 free-ranging vicuñas) and 614 domestic South American camelids (571 alpacas and 43 llamas), in ten Andean communities at the Salinas y Aguada Blanca reserve, province of Arequipa, southern Peru. Samples were tested for the presence of leptospirosis, foot and mouth disease (FMD), bovine viral diarrhea (BVD), bovine herpesvirus type 1 (BHV-1), brucellosis, bluetongue disease (BT), paratuberculosis, and neosporosis. Serological results showed that 1.9% (4/207) of vicuñas, 18.6% (106/571) of alpacas, and 23.3% (10/43) of llamas were positive to one or more Leptospira serovars. One percent of vicuñas (2/207) and 2.4% of domestic camelids (15/614) had Neospora caninum antibodies tested by ELISA, but only two vicuñas and two alpacas were confirmed by Western blot. Epidemiological evaluation found an association of leptospirosis to sex and age (p < 0.001), with female subjects older than 2.5 years at higher risk of infection. Interestingly, antibodies against Leptospira serovars were only found in captive vicuñas. This is the first study where health status of free-ranging and captive vicuñas has been compared. Results indicate minimal to nil presence of FMD, BVD, BHV-1, brucellosis, BT, paratuberculosis, and neosporosis allied to health disorders in our sample. The detection of seropositive animals against Leptospira, however, unveils the likely significance of leptospirosis in wild and domestic South American camelids, the impact of mixed husbandry over vicuña population and the risk to human health.