RESUMO
Domestic birds such as Gallus gallus, Meleagris gallopavo, Anser anser and Numida meleagris are widely distributed throughout the world and maintain contact with humans and other animal species considered reservoirs of both Visceral Leishmaniasis (VL) and American Tegumentary Leishmaniasis (ATL), including dogs and cats; wild canids, marsupials; and synanthropic animals such as rodents and chiroptera. Therefore, this study aimed to detect the presence of anti-Leishmania spp. antibodies in birds from a rural area of the municipality of Santa Maria, southern Brazil. From May to December 2022, 262 blood samples were collected from 244 chickens, 8 turkeys, 7 guinea fowl and 3 geese, distributed in 27 rural properties in 6 districts. All the sites visited presented positive birds for the presence of Leishmania spp. Thus, it is inferred that, contact with this protozoan can induce the production of antibodies, suggesting that these animals can be used as sentinels for the circulation of this agent. In addition, the blood of these animals is a preferred food source for insects of the subfamily Phlebotominae, which can be used them as bioindicators of the presence of these phlebotomes.
Assuntos
Canidae , Doenças do Gato , Doenças do Cão , Leishmania , Leishmaniose Cutânea , Humanos , Animais , Cães , Gatos , Aves Domésticas , Brasil , Doenças do Gato/parasitologia , Galinhas , Doenças do Cão/parasitologia , Leishmaniose Cutânea/veterinária , GansosRESUMO
Bovine trypanosomosis, caused by Trypanosoma vivax, is a disease that originated in Africa and currently affects cattle in several South American countries, including almost all Brazilian states. Despite the reports on T. vivax infection in southern Brazil, data on its circulation status is currently unavailable. In this study, we aimed to detect anti-Trypanosoma spp. IgG antibodies in cattle from Rio Grande do Sul and suggest areas with T. vivax transmission risk. A total of 691 serum samples from cattle in the intermediate regions of Rio Grande do Sul were analyzed using indirect immunofluorescence assay (IFA). The overall seroprevalence of anti-Trypanosoma antibodies in cattle was 24.6% (170/691). The detection rate ranged from 0-37.3%, with a high prevalence in the intermediate regions of Ijuí (37.3%), Uruguaiana (30.7%), and Passo Fundo (28.9%). Thus, these regions were suggested as possible bovine trypanosomosis risk areas due to the high seroprevalence. This is the first serological study to determine Trypanosoma spp. infection status in cattle from Rio Grande do Sul, providing data on the epidemiology of trypanosomosis in the state.
Assuntos
Doenças dos Bovinos , Trypanosoma , Tripanossomíase Bovina , Tripanossomíase , Bovinos , Animais , Brasil/epidemiologia , Estudos Soroepidemiológicos , Tripanossomíase/epidemiologia , Tripanossomíase/veterinária , Tripanossomíase Bovina/diagnóstico , Tripanossomíase Bovina/epidemiologia , Tripanossomíase Bovina/parasitologia , Trypanosoma vivax , Doenças dos Bovinos/diagnóstico , Doenças dos Bovinos/epidemiologiaRESUMO
Toxoplasmosis is a parasitic disease caused by infection with the protozoan Toxoplasma gondii. In 2018, the first cases of people with clinical signs of acute febrile syndrome were reported, and in the same year, the largest outbreak of human toxoplasmosis ever described in the literature was reported. In this sense, the present work sought to describe the evolution of the outbreak cases in the municipality of Santa Maria, Rio Grande do Sul State, Brazil, as well as the studies conducted and published during and after the outbreak in the municipality (the period between 2018 and 2023). In addition, the discussion of public policies and their modifications after the notification of this outbreak. As a result of this research, verifying the evolution of notified and confirmed cases, the possibility of detection and genotypic characterization of T. gondii and the possibility of co-infections and evaluation of the humoral response is possible. With regard to public policies, the importance of detecting the agent through the heel prick test and increasing the monitoring of water quality to prevent outbreaks.
Assuntos
Toxoplasma , Toxoplasmose , Humanos , Brasil/epidemiologia , Anticorpos Antiprotozoários , Toxoplasmose/parasitologia , Toxoplasma/genética , Surtos de DoençasRESUMO
Toxoplasmosis affects various organisms, including humans. In 2018, the largest outbreak of human toxoplasmosis described so far was reported in southern Brazil, with 809 human cases reported, and water as the potentially primary source of infection. Therefore, in this study, we aimed to evaluate the seroprevalence of Toxoplasma gondii in naturally infected domestic cats before and after the human toxoplasmosis outbreak, as well as the potential for environmental contamination by the number of cats infected after the outbreak. We evaluated 381 serum samples from domestic cats in southern Brazil, using an indirect immunofluorescence assay, with samples considered positive at a titer of 1:20. We found that 73% (204/279) and 27% (75/279) of the samples analyzed before the outbreak were negative and positive, respectively. After the outbreak, 62% (69/112) were negative of the samples were and 38% (43/112) were positive. Notably, the proportion of positive samples before the outbreak before (27%) was significantly lower than that after the outbreak (38%; P = 0.020). Therefore, the increased seroprevalence of T. gondii in cats was probably correlated with the ingestion of contaminated water. Therefore, it is important to monitor animals, mainly definitive hosts, after toxoplasmosis outbreaks, considering that these animals can contaminate the environment and, consequently, humans.
Assuntos
Toxoplasma , Toxoplasmose , Humanos , Gatos , Animais , Estudos Soroepidemiológicos , Anticorpos Antiprotozoários , Surtos de Doenças/veterinária , Água , Toxoplasmose/epidemiologiaRESUMO
Toxoplasmosis is a worldwide disease caused by Toxoplasma gondii, which can infect diverse hosts, including dogs. Although T. gondii infection in dogs is usually subclinical, they are susceptible to infection and develop a specific immune response to the parasite. In 2018, the largest outbreak of human toxoplasmosis in the world occurred in Santa Maria, in southern Brazil; however, the impact of this outbreak on other hosts was not investigated at the time. Considering that dogs often share the same environmental sources of infection as humans, mainly water sources, and that in Brazil, the detection rates of anti-T. gondii immunoglobulin G (IgG) in dogs is very high, this study investigated the frequency of anti-T. gondii IgG in dogs in Santa Maria before and after the outbreak. A total of 2.245 serum samples were analyzed, 1159 collected before the outbreak and 1086 collected after the outbreak. Serum samples were tested for anti-T. gondii antibodies using an indirect immunofluorescence antibody test (IFAT). The infection detection of T. gondii was 16% (185/1159) before the outbreak and 43% (466/1086) after the outbreak. These results showed the infection of dogs with T. gondii and the high frequency of anti-T. gondii antibodies in dogs after the outbreak in humans in 2018, reinforcing water as a possible source of infection and the importance of including toxoplasmosis in the differential diagnosis of dogs.
Assuntos
Toxoplasmose , Humanos , Cães , Animais , Brasil/epidemiologia , Estudos Soroepidemiológicos , Toxoplasmose/epidemiologia , Anticorpos Antiprotozoários , Imunoglobulina G , Surtos de Doenças , Fatores de RiscoRESUMO
Parasites of the genus Sarcocystis can infect several species of animals and cause multiple diseases such as equine protozoal myeloencephalitis. Felines are considered hosts of this protozoa; therefore, the present study aimed to detect anti-Sarcocystis spp.-specific antibodies in domestic cats that were under clinical evaluation, using the indirect immunofluorescence antibody test. Anti-Sarcocystis-specific immunoglobulin Gs were detected in 24 out of 497 (4.82%) cat serum samples. These findings support the fact that natural Sarcocystis infections do occur in cats. Furthermore, it highlights the importance of domestic cats as both intermediate and definitive hosts in the Sarcocystis life cycle, maintaining the parasite and serving as a source of infection for various other animals. To the best of our knowledge, this is the first study to identify antibodies against the genus Sarcocystis in cats from a region in southern Brazil.
Assuntos
Sarcocystis , Sarcocistose , Animais , Gatos , Cavalos , Sarcocistose/veterinária , Sarcocistose/parasitologia , Brasil , Anticorpos Antiprotozoários , Técnica Indireta de Fluorescência para Anticorpo/veterináriaRESUMO
PURPOSE: The inspection of animal products is important for controlling parasitic zoonoses. Some processes that guarantee food safety to consumers such as carcass condemnation cause economic losses. This study aimed to detect Sarcocystis cysts in cattle hearts obtained from slaughterhouses and to evaluate sarcocyst viability after freezing treatment. METHODS: When myocardial tissues were minced and subjected to fresh examination, sarcocysts were observed in all analyzed tissues resulting in 21.73 cysts/g of tissue. Sarcocyst viability was verified after tissue freezing at 35 ± 2 °C and - 20 ± 2 °C for 0-12 h. After freezing, the tissues were minced, and sarcocysts were collected and stained with Tripan Blue. In addition, cysts were mechanically disrupted to check bradyzoite viability. RESULTS: Cysts and bradyzoites were unviable at - 35 °C for ≥ 3 h and - 20 °C for ≥ 8 h. CONCLUSION: These results suggest freezing treatment as an alternative to condemnation of cattle carcasses contaminated with Sarcocystis spp. Similar studies using freezing treatment with other animals infected by Sarcocystis must be conducted.
Assuntos
Sarcocystis , Sarcocistose , Animais , Bovinos , Sarcocistose/veterinária , Sarcocistose/parasitologia , Congelamento , Coração , ZoonosesRESUMO
This study aimed to detect the occurrence of infection by Leishmania spp.in bats from 34 municipalities of Rio Grande do Sul state (RS; southern Brazil) from 2016 to 2021. A total of 109 bats were provided by the Centro Estadual de Vigilância em Saúde of RS, including six species belonged to Molossidae family, six to Vespertilionidae family, and two to Phyllostomidae family. Leishmania spp. was identified using the nested-PCR method by amplifying the SSU rDNA ribosomal subunit gene into four organ pools: (1) the liver, spleen, and lymph node; (2) heart and lungs; (3) skin; and (4) bone marrow of each bat. Three (3/109, 2.7%) animals tested positive for Leishmania spp. The respective PCR-positive organs came from pools 1 and 3. Two bats (Tadarida brasiliensis) were from the municipality of Canoas, and sequences analysis confirms the species identification as Leishmania infantum. In the third bat (Molossus molossus), from Rio Grande, it was not possible to determine the protozoa species, being considered Leishmania spp. Our results indicate that bats can participate in the biological cycle of Leishmania spp. and perform as host, reservoir, and/or source of infection of the protozoa in different areas of RS. More studies will be needed to elucidate the role of these Chiropteras in the circulation of Leishmania spp. This is the first study reporting the occurrence of Leishmania spp. in bats in Rio Grande do Sul state, southern Brazil.
Assuntos
Quirópteros , Leishmania infantum , Animais , Brasil/epidemiologia , Quirópteros/parasitologia , DNA Ribossômico/genética , Leishmania infantum/genética , PrevalênciaRESUMO
Toxoplasmosis is a disease caused by T. gondii, a protozoa which affects humans and animals and is widely distributed worldwide. In humans, there is great concern due to the serious consequences that can occur in the infection of pregnant women and the newborn. The early diagnosis of gestational toxoplasmosis is important for treatment to be carried out in order to prevent vertical transmission or reduce damage. The diagnosis can be made through the detection of antibodies in pregnant women or neonates and PCR of amniotic fluid. Previous studies have also reported PCR of the placenta as a good diagnostic test. Our study evaluated the detection of T. gondii DNA in placenta samples from parturients seen at the University Hospital of Santa Maria, Southern Brazil and treated during the pregnancy. We performed PCR in forty samples and five were positive, representing 12.5%. When correlating the treatment time and the detection of DNA in the placentas, no significant result was found. The prevalence of positive samples was lower than in other studies in the literature. The data reaffirm the importance of carrying out the analysis of the placenta.
Assuntos
Placenta/parasitologia , Complicações Parasitárias na Gravidez/epidemiologia , Toxoplasma/isolamento & purificação , Toxoplasmose/epidemiologia , Adulto , Brasil/epidemiologia , Feminino , Humanos , Gravidez , Complicações Parasitárias na Gravidez/parasitologia , Prevalência , Toxoplasmose/parasitologia , Adulto JovemRESUMO
Search to a new alternative to control bovine ticks (Rhipicephalus microplus), the present study aimed to evaluate the acaricidal activity of organo-modified siloxane alone and in association with different commercial products or with piperonyl butoxide (BPO). Engorged females were subjected to an in vitro immersion test and 10 groups were used: control, 0.5% siloxane, 1% siloxane, 2% siloxane, 0.5% siloxane + 5% BPO, 1% siloxane + 5% BPO, 2% siloxane + 5% BPO, commercial product, 0.5% siloxane + commercial product, and 1% siloxane + commercial product. After immersion, engorged females were incubated for 14 days for oviposition and hatchability tests. Another immersion test was performed with 5% siloxane and 2.5% siloxane + 10% BPO to evaluate the histopathological changes. Then, engorged females were incubated for 0, 2, 4, 8, 12, 24, 36, 48, 60, 72, 84, 96, 120, 144, and 168 h and immersed in 10% formaldehyde for later analyses. The unassociated siloxane showed an acaricide efficacy of 93.88% at 2% concentration, and when associated with 5% BPO, it reached 100% efficacy at all tested concentrations. The tested commercial products showed enhanced efficacy when associated with siloxane. Histopathological analysis showed cell changes in both treatments and total cell disintegration after 120 h in the 5% siloxane group and after 96 h in the 2.5% siloxane + 10% BPO group. Therefore, siloxane alone or in combination is an alternative against R. microplus, and siloxane enhances the efficacy of available commercial products.
Assuntos
Acaricidas , Doenças dos Bovinos , Rhipicephalus , Infestações por Carrapato , Acaricidas/farmacologia , Animais , Bovinos , Feminino , Larva , SiloxanasRESUMO
The protozoan Toxoplasma gondii is a causative agent of toxoplasmosis, an important and widespread zoonotic disease. The transmission of this disease in humans includes ingestion of sporulated oocysts present in contaminated water or food. T. gondii oocysts are widely distributed and toxoplasmosis is considered a major food- and waterborne pathogen worldwide, making drinking water containing sporulated T. gondii oocysts a major source of contamination for people. In the first half of 2018, an unprecedented outbreak of toxoplasmosis was reported in the city of Santa Maria, southern Brazil. The temporal and spatial distribution of the cases strongly suggested a waterborne infection. Thus, the aim of this study was to investigate a possible involvement of treated water as a source of the outbreak. For this, piglets received potentially contaminated water ad libitum for 21 days and the infection was monitored by serology through IFAT and investigation of T. gondii DNA in tissues by PCR amplification of a 529 bp followed by mouse bioassays. All piglets receiving test water ad libitum for 21 days as well as positive controls seroconverted to T. gondii. T. gondii DNA was detected in 62.5% of the piglets that received test water. All mice inoculated with tissues from each positive piglet were PCR-positive. These results strongly indicated the presence of viable oocysts in the test water administered to the animals during the study.
Assuntos
Bioensaio , Oocistos , Toxoplasma/isolamento & purificação , Toxoplasmose/epidemiologia , Poluição da Água , Água/parasitologia , Animais , Brasil/epidemiologia , Surtos de Doenças , Humanos , Suínos , Toxoplasma/genética , Toxoplasmose/parasitologiaRESUMO
Giardiasis is an important and prevalent zoonosis in dogs and humans caused by Giardia spp. The close relationship between pets and humans has physical, emotional and social benefits. The dogs have an important role in Giardia duodenalis cycle and transmission. This study aimed to verify the occurrence of the parasite in dogs from Central Region, in Santa Maria, Rio Grande do Sul State, Brazil, from April to October 2018. Dog feces (230) were submitted to Faust coproparasitological and molecular analyses. The positive samples in the nested-PCR (β-giardin gene) were sent for DNA sequencing and phylogenetic analyses (Neighbor-Joining). The occurrence of G. duodenalis, was 5.6% (13/230) and 4.3% (10/230) detected by coproparasitological technique and nested-PCR, respectively. There was no difference in the sensitivity of the tests used. From the faecal samples analyzed, there were no differences among the variables: diagnostic techniques, local, sex, and age of the animals (p>0.05). Only in the stool examination methodology a difference was observed between the ages (p<0.05). G. duodenalis assemblages were C and D, frequently reported in dogs. The close relationship between dogs and people may allow co-infections of circulating parasites in the population, including Giardia spp. and increasing the risk of transmission of zoonotic agents.(AU)
A giardíase é uma zoonose importante e prevalente em cães e humanos, sendo causada por Giardia spp. A estreita relação entre animais de estimação e seres humanos traz benefícios físicos, emocionais e sociais. Os cães têm um papel importante no ciclo e transmissão de Giardia duodenalis. Este estudo teve como objetivo verificar a ocorrência do parasita em cães da Região Central, em Santa Maria, RS, Brasil, de abril a outubro de 2018. As fezes de cães (230) foram submetidas a técnica coproparasitológica de Faust e análises moleculares. As amostras positivas no nested-PCR (gene β-giardin) foram enviadas para sequenciamento de DNA e posterior análise filogenética (Neighbor-Joining). A ocorrência de G. duodenalis foi de 5,6% (13/230) e 4,3% (10/230) detectados pela técnica coproparasitológica e nested-PCR, respectivamente. Não houve diferença na sensibilidade dos testes utilizados. Das amostras fecais analisadas, não houve diferenças entre as variáveis: técnicas de diagnóstico, local, sexo e idade dos animais (p>0,05). Somente na metodologia de exame de fezes observou-se diferença entre as idades (p<0,05). As assemblages de G. duodenalis encontradas foram C e D, frequentemente relatadas em cães. A estreita relação entre cães e pessoas pode permitir co-infecções de parasitas circulantes na população, incluindo Giardia spp. e aumentando o risco de transmissão de agentes zoonóticos.(AU)
Assuntos
Animais , Cães , Filogenia , Reação em Cadeia da Polimerase , Giardíase , Cães/parasitologia , Animais de Estimação , GiardiaRESUMO
ABSTRACT: Giardiasis is an important and prevalent zoonosis in dogs and humans caused by Giardia spp. The close relationship between pets and humans has physical, emotional and social benefits. The dogs have an important role in Giardia duodenalis cycle and transmission. This study aimed to verify the occurrence of the parasite in dogs from Central Region, in Santa Maria, Rio Grande do Sul State, Brazil, from April to October 2018. Dog feces (230) were submitted to Faust coproparasitological and molecular analyses. The positive samples in the nested-PCR (-giardin gene) were sent for DNA sequencing and phylogenetic analyses (Neighbor-Joining). The occurrence of G. duodenalis, was 5.6% (13/230) and 4.3% (10/230) detected by coproparasitological technique and nested-PCR, respectively. There was no difference in the sensitivity of the tests used. From the faecal samples analyzed, there were no differences among the variables: diagnostic techniques, local, sex, and age of the animals (p>0.05). Only in the stool examination methodology a difference was observed between the ages (p 0.05). G. duodenalis assemblages were C and D, frequently reported in dogs. The close relationship between dogs and people may allow co-infections of circulating parasites in the population, including Giardia spp. and increasing the risk of transmission of zoonotic agents.
RESUMO: A giardíase é uma zoonose importante e prevalente em cães e humanos, sendo causada por Giardia spp. A estreita relação entre animais de estimação e seres humanos traz benefícios físicos, emocionais e sociais. Os cães têm um papel importante no ciclo e transmissão de Giardia duodenalis. Este estudo teve como objetivo verificar a ocorrência do parasita em cães da Região Central, em Santa Maria, RS, Brasil, de abril a outubro de 2018. As fezes de cães (230) foram submetidas a técnica coproparasitológica de Faust e análises moleculares. As amostras positivas no nested-PCR (gene -giardin) foram enviadas para sequenciamento de DNA e posterior análise filogenética (Neighbor-Joining). A ocorrência de G. duodenalis foi de 5,6% (13/230) e 4,3% (10/230) detectados pela técnica coproparasitológica e nested-PCR, respectivamente. Não houve diferença na sensibilidade dos testes utilizados. Das amostras fecais analisadas, não houve diferenças entre as variáveis: técnicas de diagnóstico, local, sexo e idade dos animais (p>0,05). Somente na metodologia de exame de fezes observou-se diferença entre as idades (p 0,05). As assemblages de G. duodenalis encontradas foram C e D, frequentemente relatadas em cães. A estreita relação entre cães e pessoas pode permitir co-infecções de parasitas circulantes na população, incluindo Giardia spp. e aumentando o risco de transmissão de agentes zoonóticos.
RESUMO
The present study aimed to describe a molecular analysis of environmental and pork samples, the isolation, genetic identification and immunohistochemistry (IHC) of Toxoplama gondii from placenta and amniotic fluid from five pregnant women that miscarried during a toxoplasmosis outbreak in 2018, Santa Maria, Rio Grande do Sul. Environmental and pork samples were submitted to polymerase chain reaction (PCR); placenta and amniotic fluid samples to histopathology, IHC, mouse bioassay and PCR. All samples were genotyped by PCR-RFLP with 11 loci. Histopathologic and IHC were compatibles with toxoplasmosis. All pregnants were positive in PCR and bioassay, the genotypes were compared, and all were equal suggesting a same source of infection. Among the environmental and food samples, a sludge sample from a water tank and two porks samples were positive in PCR, and the genotypes were different from the pregnant women isolates. It is concluded that obtain and compare isolates is essential to elucidate outbreak source.
Assuntos
Surtos de Doenças , Placenta/parasitologia , Complicações na Gravidez , Toxoplasma/classificação , Toxoplasma/genética , Toxoplasmose/epidemiologia , Toxoplasmose/parasitologia , Brasil/epidemiologia , Suscetibilidade a Doenças , Meio Ambiente , Feminino , Humanos , Gravidez , Vigilância em Saúde Pública , Toxoplasma/isolamento & purificação , Toxoplasmose/diagnósticoRESUMO
Reproductive tests in cattle are of great economic importance, given the impact it can have on the production system and may be caused by agents. Neospora caninum and Bovine Viral Diarrhea virus (BVDV) are considered of great importance as reproductive and should be considered responsible for keeping animals persistently infected. The present study included 479 calf serum samples for export in the state of Rio Grande do Sul (RS). All samples were screened for BVDV by an ELISA antigen. BVDV antigen-positive ELISA samples were isolated from BVDV in cell culture. An indirect immunofluorescence (IFT) technique was used to detect anti-N. caninum antibodies. Of the 479 export-treated serum samples, 361 were positive for BVDV antigens by ELISA and/or viral isolation test (361/479-75.36%), and 109 IFT-positive samples for N. caninum (109/479-22.75%). Despite detection of antibodies anti-N. caninum did not differ statistically between naturally infected BVDV and non-BVDV infected animals suggesting that there is no interference of BVDV infection on infection or detection rate of animals with N. caninum, positive animals in viral isolation and high DO in BVDV-Ag ELISA. may present active disease and consequent immunosuppression, contributing to a potential reactivation of N. caninum.(AU)
Testes reprodutivos em bovinos são de grande importância econômica, dado o impacto que podem ter no sistema de produção e podem ser causados por agentes. O Neospora caninum e o vírus da Diarreia Viral Bovina (BVDV) são considerados de grande importância como reprodutivos e devem ser considerados responsáveis por manter os animais persistentemente infectados. O presente estudo incluiu 479 amostras de soro de bezerro para exportação no estado do Rio Grande do Sul (RS). Todas as amostras foram rastreadas para BVDV por um antígeno ELISA. As amostras de ELISA positivas para o antigénio BVDV foram isoladas a partir de BVDV em cultura de células. Uma técnica de imunofluorescência indireta (IFT) foi utilizada para detectar anticorpos anti-N caninum. Das 479 amostras de soro tratadas para exportação, 361 foram positivas para antígenos de BVDV por ELISA e/ou teste de isolamento viral (361/479-75,36%) e 109 amostras positivas para IFT para N. caninum (109/479-22,75%). Apesar da detecção de anticorpos anti-N. caninum não diferiu estatisticamente entre animais infectados naturalmente BVDV e não BVDV sugerindo que não há interferência da infecção pelo BVDV na infecção ou taxa de detecção de animais com N. caninum, animais positivos em isolamento viral e alta DO em BVDV-Ag ELISA, pode apresentar doença ativa e consequente imunossupressão, contribuindo para uma potencial reativação de N. caninum.(AU)
Assuntos
Animais , Bovinos , Coccidiose/veterinária , Vírus da Diarreia Viral Bovina/isolamento & purificação , Neospora/isolamento & purificação , Coinfecção/veterinária , Coinfecção/epidemiologiaRESUMO
Molecular detection of Eimeria species in fecal samples can be useful for experimental and diagnostic purposes. However, the parasite quantity presence in feces and the oocyst wall are an obstacle in DNA extraction protocols. Therefore, adequate sampling and effective disruption of the oocysts are essential to improve the accuracy of DNA detection by PCR. The aims of this study were to evaluate the suitability of six protocols for DNA extraction from Eimeria spp. present in bovine and sheep. Twenty pools of fecal samples from cattle (10 pools) and sheep (10 pools) were distributed to six DNA extraction protocols: commercial kit, commercial kit with modification, DNAzol, cetyl-trimethyl ammonium bromide (CTAB), glass beads and commercial kit for fecal samples. Fecal samples were submitted to DNA extraction and PCR. Among the protocols tested, CTAB was determined to be most suitable for DNA extraction from oocysts (90% of DNA detection by PCR); DNAzol and CTAB resulted in higher DNA detection from bovine samples (80%). CTAB and commercial kit with modification improved PCR detection of Eimeria spp. in sheep samples, with positive amplification of DNA in all tested samples.(AU)
A detecção molecular de espécies de Eimeria em amostras fecais pode ser útil para fins experimentais e de diagnóstico. No entanto, a quantidade de parasitas nas fezes e a parede do oocisto são um obstáculo nos protocolos de extração de DNA. Portanto, uma amostragem adequada e a ruptura efetiva dos oocistos são essenciais para melhorar a precisão da detecção de DNA por PCR. Os objetivos deste estudo foram avaliar seis protocolos para extração de DNA de Eimeria spp. em amostras de bovinos e ovinos. Foram distribuídos 20 grupos de amostras fecais de bovinos (10 grupos) e ovinos (10 grupos) em seis protocolos de extração de DNA: kit comercial, kit comercial com modificação, DNAzol, brometo de cetil-trimetil amônio (CTAB), pérolas de vidro e kit comercial para amostras fecais. As amostras fecais foram submetidas à extração de DNA e PCR. Entre os protocolos testados, CTAB foi considerado o mais adequado para extração de DNA de oocistos (90% de detecção de DNA por PCR); DNAzol e CTAB resultaram em maior detecção de DNA em amostras de bovinos (80%). CTAB e kit comercial com modificação melhoraram a detecção por PCR de Eimeria spp. em amostras de ovinos, amplificação positiva de DNA em todas as amostras testadas.(AU)
Assuntos
Animais , Bovinos , Doenças dos Ovinos/parasitologia , Doenças dos Bovinos/parasitologia , Coccidiose/diagnóstico , Coccidiose/veterinária , Eimeria/genética , Reação em Cadeia da Polimerase/veterinária , OocistosRESUMO
Serological techniques can detect antibodies against Sarcocystis spp., Neospora caninum and Toxoplasma gondii antigens in single or mixed infections. Immunofluorescent antibody tests (IFAT) is considered the gold standard technique for Sarcocystosis diagnostic in cattle serum and a positive IFAT result reflects Sarcocystis spp. infection. Therefore, the aims of the present study were to compare IFAT and Dot-blot for sarcocystosis diagnostic in experimentally infected mice and to investigate serological cross-reactions with N. caninum and T. gondii in these methods. Mice (Mus musculus) were inoculated intraperitoneally with bradizoites of Sarcocystis spp. or tachyzoites of N. caninum or T. gondii. Serum samples were obtained and analyzed by IFAT and Dot-blot for the three protozoa. Serum from N. caninum and T. gondii experimentally infected mice were tested by IFAT and reacted only to N. caninum or T. gondii antigens, respectively. Specific antibodies against Sarcocystis spp. were present in all animals experimentally infected with this protozoan, with IFAT titers from 10 to 800. Serum samples from mice experimentally infected with Sarcocystis spp., N. caninum and T. gondii and tested by Dot-blot demonstrated no cross reaction between protozoa. A Dot-blot using Sarcocystis spp. antigen appears to be a good alternative to IFAT in the serological diagnosis of Sarcocystosis.(AU)
As técnicas sorológicas podem detectar anticorpos contra os antígenos de Sarcocystis spp., Neospora caninum e Toxoplasma gondii em infecções únicas ou mistas. O teste de anticorpos imunofluorescentes (IFAT) é considerado a técnica padrão-ouro para o diagnóstico de sarcocistose no soro de bovinos e um resultado positivo de IFAT reflete Sarcocystis spp. infecção. Portanto, os objetivos do presente estudo foram comparar IFAT e Dot-blot para diagnóstico de sarcocistose em camundongos infectados experimentalmente e investigar reações cruzadas sorológicas com N. caninum e T. gondii nesses métodos. Os camundongos (Mus musculus) foram inoculados intraperitonealmente com bradizoítos de Sarcocystis spp. ou taquizoítos de N. caninum ou T. gondii. As amostras de soro foram obtidas e analisadas por IFAT e Dot-blot para os três protozoários. O soro de N. caninum e T. gondii infectados experimentalmente foram testados por IFAT e reagiram apenas aos antígenos de N. caninum ou T. gondii, respectivamente. Anticorpos específicos contra Sarcocystis spp. estavam presentes em todos os animais experimentalmente infectados com este protozoário, com títulos de IFAT de 10 a 800. Amostras de soro de camundongos infectados experimentalmente com Sarcocystis spp., N. caninum e T. gondii e testadas por Dot-blot não demonstraram reação cruzada entre protozoários. Um Dot-blot usando Sarcocystis spp. O antígeno parece ser uma boa alternativa ao IFAT no diagnóstico sorológico da sarcocistose.(AU)
Assuntos
Animais , Masculino , Camundongos , Bovinos/parasitologia , Testes Sorológicos/métodos , Doenças dos Bovinos , Sarcocystis , Sarcocistose/diagnóstico , Sarcocistose/veterinária , Testes Sorológicos/veterinária , Técnica Indireta de Fluorescência para AnticorpoRESUMO
Toxoplasma gondii is a protozoan that has great genetic diversity and is prevalent worldwide. In 2018, an outbreak of toxoplasmosis occurred in Santa Maria, Brazil, which was considered the largest outbreak ever described in the world. This paper describes the isolation and molecular characterization of Toxoplasma gondii from the placenta of two pregnant women with acute toxoplasmosis who had live births and were receiving treatment for toxoplasmosis during the outbreak. For this, placental tissue samples from two patients underwent isolation by mice bioassay, conventional PCR and genotyping using PCR-RFLP with twelve markers. Both samples were positive in isolation in mice. The isolate was lethal to mice, suggesting high virulence. In addition, the samples were positive in conventional PCR and isolates submitted to PCR-RFLP genotyping presented an atypical genotype, which had never been described before. This research contributes to the elucidation of this great outbreak in Brazil.
Assuntos
Coccidiostáticos/uso terapêutico , Placenta/parasitologia , Complicações Infecciosas na Gravidez/tratamento farmacológico , Toxoplasma/genética , Toxoplasmose/tratamento farmacológico , Animais , Brasil/epidemiologia , Modelos Animais de Doenças , Surtos de Doenças , Feminino , Genótipo , Humanos , Leucovorina/uso terapêutico , Nascido Vivo , Camundongos , Polimorfismo de Fragmento de Restrição , Gravidez , Complicações Infecciosas na Gravidez/epidemiologia , Complicações Infecciosas na Gravidez/parasitologia , Pirimetamina/uso terapêutico , Sulfadiazina/uso terapêutico , Toxoplasma/isolamento & purificação , Toxoplasmose/epidemiologia , Toxoplasmose/parasitologiaRESUMO
BACKGROUND: Reports of a lack of efficacy of most of the anthelmintic compounds for ruminants associated with the long-time necessity for creating new molecules have stressed the urgency to adopt alternative methods to control gastrointestinal parasites infection, such as strategies of sharing grazing areas. Therefore, this study aimed to evaluate nematode populations affecting cattle and sheep that share grazing areas before and after treatment with different anthelmintic compounds, and investigate the efficacy of anthelmintic treatment in these naturally infected ruminants at farms in the state of Rio Grande do Sul, Brazil. METHODS: The presence of co-infections by Haemonchus species was investigated by polymerase chain reaction (PCR) for groups treated with a benzimidazole. Farms were selected by: farmers' consent, presence of 42-60 (or more) calves and sheep per farm with counts of ≥ 200 eggs per gram of feces (EPG), availability of calves and lambs aging from 6 to 9 months, absence of anthelmintic treatment for both species for 60 days before the experimental period, and shared grazing areas between this species on each farm. Animals were distributed into six treatment groups for each ruminant species per farm and treated with: ivermectin, doramectin, moxidectin, levamisole, albendazole, and closantel. RESULTS: Levamisol was the most effective anthelmintic compound for both ruminant species. In general, Cooperia spp., Haemonchus spp., and Trichostrongylus spp. were the genus present after tested treatments that were ineffective. PCR showed the presence of Haemonchus species co-infections between cattle and sheep. CONCLUSION: Therefore, this study demonstrated the similarity between nematode population, the presence of multi-resistant nematodes, and the presence of Haemonchus species co-infections affecting different ruminant species that share pastures.