RESUMO
The transplacental transmission of parasites and hemoparasites is crucial for understanding the epidemiology of diseases. This study aimed to assess the prevalence of hemopathogens in bovine fetuses at various gestational periods. Samples were obtained from a slaughterhouse in the state of Minas Gerais, Brazil, and a total of 236 fetuses were collected. DNA extracted from blood samples (145) and organ samples (a pool of brain and spleen) (236) underwent a nested PCR (nPCR) assay to detect Babesia spp., Theileria spp., Trypanosoma vivax, Anaplasma marginale, Anaplasma bovis, Anaplasma phagocytophilum, Ehrlichia minasensis, and hemotropic Mycoplasma spp. Additionally, serological analysis of 145 plasma samples was conducted using the indirect fluorescent antibody test-IFAT to detect IgG against Babesia bovis, Babesia bigemina, A. marginale, and Trypanosoma vivax. The observed prevalence of transplacental transmission was 19.3 %, 6.2 %, 42.7 % and 2.7 %, for A. marginale, B. bigemina, 'Candidatus M. haemobos', and Mycoplasma wenyonii, respectively. The prevalence of A. marginale by gestational trimester was 16 % (13/81) in the second trimester and 23 % (14/60) in the third trimester, with no positive samples in the first trimester. Regarding the species B. bovis and B. bigemina, all evaluated animals tested negative by nPCR, and no serological evidence for B. bovis was found by the IFAT. Babesia bigemina demonstrated an overall seroprevalence of 6.2 % (9/145), with 4.8 % (7/145) in the last trimester and 1.3 % (2/145) in the second trimester of pregnancy. In total, 42.7 % (62/145) of blood samples were positive for 'Candidatus M. haemobos', with 42 % (34/81) in the middle trimester, and 43 % (26/60) in the final trimester of pregnancy. Mycoplasma wenyonni was detected in 2.7 % (4/145) blood samples, all in coinfection with 'C. M. haemobos'. The prevalence by pregnancy trimester was 25 % (1/4) in the first trimester; 1.2 % (1/81) in the second trimester and 3.3 % (2/60) in the third trimester of pregnancy. Hemopathogen DNA was detected in fetus blood samples but not the brain or spleen samples. All the samples were negative for T. vivax, Theileria spp., Anaplasma spp. and Ehrlichia spp. Overall, in this study, approximately 70 % of fetuses were positive for one or more of the studied parasites. No significant associations were observed between pairs of pathogens, except 'C. M. haemobos' and A. marginale.
Assuntos
Doenças dos Bovinos , Mycoplasma , Animais , Brasil/epidemiologia , Bovinos , Feminino , Doenças dos Bovinos/epidemiologia , Doenças dos Bovinos/microbiologia , Doenças dos Bovinos/parasitologia , Mycoplasma/isolamento & purificação , Gravidez , Prevalência , Babesia/isolamento & purificação , Feto/microbiologia , Feto/parasitologia , Infecções por Mycoplasma/epidemiologia , Infecções por Mycoplasma/veterinária , Infecções por Mycoplasma/microbiologia , Theileria/isolamento & purificação , Trypanosoma vivax/isolamento & purificação , Transmissão Vertical de Doenças Infecciosas/veterinária , Anaplasma/isolamento & purificação , Babesiose/epidemiologia , Babesiose/parasitologia , Anaplasmose/epidemiologia , Anaplasmose/microbiologia , Ehrlichia/isolamento & purificaçãoRESUMO
The reintroduction of captive animals to the wild helps restore endangered species, but it risks pathogen transmission, harming wild populations. Such transmission can impact the genetic diversity and long-term viability of these populations. This study assessed parasite diversity and load in captive Pecari tajacu, a species native to the Americas and culturally significant to Brazilian indigenous culture, prior to reintroduction. Samples from 24 peccaries were analyzed for ectoparasites, hemopathogens, and stool parasites with direct and molecular analysis. Findings showed that various parasites were present. Two peccaries (8.3%) were infested by the adult tick Amblyomma sculptum. Six (25.0%) tested positive for Trypanosoma evansi, four (16.7%) for hemobacteria of the family Anaplasmataceae, twelve (50.0%) for hemotropic Mycoplasma, and seven (29.2%) for Leishmania braziliensis. Stool samples indicated multiple parasites, with sixteen (66.7%) peccaries infected by Strongylida order parasites, Spiruridae in three (12.5%), and Ascaris suum in one (4.2%) animal. Cysts of Balantidium sp. were found in twenty (83.3%), Entamoeba polecki in five (20.8%), and Iodamoeba bütschlii in two (8.3%) peccaries. To our current knowledge, this is the first global report of Leishmania braziliensis, Iodamoeba bütschlii, and Entamoeba polecki in P. tajacu, irrespective of the environment, including both captivity and wild conditions. Some of these parasites are common in domestic animals, and others are zoonotic, indicating potential interspecies pathogen transmission.
RESUMO
The zoonotic virus SARS-CoV-2, which causes severe acute respiratory syndrome in humans (COVID-19), has been identified in cats. Notably, most positive cases were in cats that had close contact with infected humans, suggesting a role for humans in animal transmission routes. Previous studies have suggested that animals with immune depletion are more susceptible to SARS-CoV-2 infection. To date, there is limited evidence of SARS-CoV-2 infections in stray and free-range cats affected by other pathogens. In this study, we investigated infections caused by SARS-CoV-2, Leishmania spp., Toxoplasma gondii, Mycoplasma spp., Bartonella spp., Feline leukemia virus (FeLV), and Feline immunodeficiency virus (FIV) in stray cats from an urban park in Brazil during the COVID-19 pandemic. From February to September 2021, 78 mixed-breed cats were tested for SARS-CoV-2 and hemopathogens using molecular analysis at Américo Renné Giannetti Municipal Park, Belo Horizonte, Minas Gerais, Brazil. An enzyme-linked immunosorbent assay (ELISA) was used to detect IgG in T. gondii. None of the animals in this study showed any clinical signs of infections. The SARS-CoV-2 virus RNA was detected in 7.7 % of cats, and a whole virus genome sequence analysis revealed the SARS-CoV-2 Delta lineage (B.1.617.2). Phylogenetic analysis showed that SARS-CoV-2 isolated from cats was grouped into the sublineage AY.99.2, which matches the epidemiological scenario of COVID-19 in the urban area of our study. Leishmania infantum was detected and sequenced in 9 % of cats. The seroprevalence of T. gondii was 23.1 %. Hemotropic Mycoplasma spp. was detected in 7.7 % of the cats, with Mycoplasma haemofelis and Candidatus Mycoplasma haemominutum being the most common. Bartonella henselae and Bartonella clarridgeiae were detected in 38.5 % of the cats, FeLV was detected in 17,9 %, and none of the cats studied tested positive for FIV. This study reports, for the first time, the SARS-CoV-2 infection with whole-genome sequencing in stray cats in southeastern Brazil and co-infection with other pathogens, including Bartonella spp. and Feline leukemia virus. Our study observed no correlation between SARS-CoV-2 and the other detected pathogens. Our results emphasize the importance of monitoring SARS-CoV-2 in stray cats to characterize their epidemiological role in SARS-CoV-2 infection and reinforce the importance of zoonotic disease surveillance.
Assuntos
COVID-19 , Doenças do Gato , Coinfecção , Vírus da Imunodeficiência Felina , Gatos , Animais , Humanos , Coinfecção/epidemiologia , Coinfecção/veterinária , Brasil/epidemiologia , Estudos Soroepidemiológicos , Pandemias , Filogenia , COVID-19/epidemiologia , COVID-19/veterinária , SARS-CoV-2/genética , Vírus da Leucemia Felina , Doenças do Gato/epidemiologiaRESUMO
Determining the occurrence of Rickettsia spp. and Anaplasma phagocytophilum in municipalities with no case records is important to define surveillance strategies and is essential to reduce lethality in different regions. Therefore, an approach aimed at enhancing surveillance in municipalities with an unknown epidemiological situation was tested, according to the classification suggested by Resolution SMA/SES 07/01/16. Canine sera collected in the annual anti-rabies campaign were submitted to the indirect fluorescent antibody test for Rickettsia amblyommatis, R. belli, R. parkeri, R. rickettsii and A. phagocytophilum. Titers ≥1:64 and ≥1:320 were considered positive for Rickettsia spp. and A. phagocytophilum, respectively. For Rickettsia spp., 61.8% of dogs were seropositive, with 26% positive for more than one species, and 42.3% were seropositive for R. rickettsii. Dogs from the urban area presented 5.16 (CI 1.18; 7.69) times greater odds of seropositivity for R. parkeri (p = 0.037) and 3.39 (CI 1.04; 3.70) times greater odds for R. belli (p = 0.017). Considering the 1:40 cutoff point, 19.1% of dogs were reactive for A. phagocytophilum. Two (1%) dogs in rural areas were positive (titer 1:640). The results indicate all species ever tested in Lavras/MG, since the present study is the city's first report on the subject. According to classifications of the aforementioned Resolution, the results determine that the municipality of Lavras should be considered a "risk area" for Brazilian spotted fever(BSF). The methodology presented is efficient, straight forward to perform and inexpensive for diagnosing a risk situation for BSF and human granulocytic anaplasmosis. Moreover, its use can be applied throughout Brazil and other countries as a public health alert guideline.
Assuntos
Anaplasma phagocytophilum , Doenças do Cão , Infecções por Rickettsia , Rickettsia , Cães , Animais , Humanos , Infecções por Rickettsia/diagnóstico , Infecções por Rickettsia/epidemiologia , Infecções por Rickettsia/veterinária , Doenças do Cão/diagnóstico , Doenças do Cão/epidemiologia , Doenças do Cão/microbiologia , Estudos SoroepidemiológicosRESUMO
Despite previous reports of SARS-CoV-2 infection in dogs and cats worldwide, the type of swab sample used for its detection through RT-qPCR needs to be better compared and described. Accordingly, as part of a multicenter study in Brazil, the aim of the present study was to assess which rectal or oropharyngeal swabs would be more appropriate for detecting SARS-CoV-2 in cats and dogs, through viral load comparison. Pets of owners diagnosed with COVID-19 in the last 7 days were eligible. A total of 148 animals from four of the five Brazilian geographical regions were analyzed, among which 10/48 cats (20.83%) and 11/100 dogs (11.00%) were positive. The results suggested that oropharyngeal swabs should be considered for SARS-CoV-2 detection, particularly in cats, due to the higher cDNA viral load. Also, the genomic results showed similarities between SARS-CoV-2 animal variants and human variants that were circulating at the time of sampling, thus corroborating the existence of zooanthroponotic transmission. In conclusion, the present study highlighted the importance of SARS-CoV-2 monitoring among cats and dogs, as virus modification may indicate the possibility of mutations in animals and spillover back to owners. Thus, positive individuals should always self-isolate from their pets during COVID-19, to prevent trans-species transmission and mutation.
Assuntos
COVID-19 , Doenças do Gato , Doenças do Cão , Humanos , Gatos , Cães , Animais , SARS-CoV-2/genética , COVID-19/diagnóstico , COVID-19/epidemiologia , COVID-19/veterinária , Brasil/epidemiologia , Doenças do Gato/diagnóstico , Doenças do Gato/epidemiologia , Doenças do Cão/diagnóstico , Doenças do Cão/epidemiologiaRESUMO
BACKGROUND: visceral leishmaniasis (VL) is a critical public health problem in over ninety countries. The control measures adopted in Brazil have been insufficient when it comes to preventing the spread of this overlooked disease. In this context, a precise diagnosis of VL in dogs and humans could help to reduce the number of cases of this disease. Distinct studies for the diagnosis of VL have used single recombinant proteins in serological assays; however, the results have been variable, mainly in relation to the sensitivity of the antigens. In this context, the development of multiepitope-based proteins could be relevant to solving such problem. METHODS: a chimeric protein (rMELEISH) was constructed based on amino acid sequences from kinesin 39 (k39), alpha-tubulin, and heat-shock proteins HSP70 and HSP 83.1, and tested in enzyme-linked immunosorbent (ELISA) for the detection of L. infantum infection using canine (n = 140) and human (n = 145) sera samples. RESULTS: in the trials, rMELEISH was able to discriminate between VL cases and cross-reactive diseases and healthy samples, with sensitivity and specificity values of 100%, as compared to the use of a soluble Leishmania antigenic extract (SLA). CONCLUSIONS: the preliminary data suggest that rMELEISH has the potential to be tested in future studies against a larger serological panel and in field conditions for the diagnosis of canine and human VL.
RESUMO
Vector-borne pathogens (VBPs) are primarily transmitted by arthropod vectors, but secondary ways of transmission have been described, including via venereal route. Nonetheless, there is still limited research on possible sexual transmission of VBPs in dogs. We molecularly investigated the presence of vector-borne pathogens in semen from dogs living in an area where these agents are endemic. Upon PCR testing, seven out of 22 (31.8%) semen samples tested positive for at least one VBP, whereas simultaneous positivity to two or more pathogens was detected in three (13.6%) dogs. Among pathogens detected in semen, Trypanosoma cruzi (n = 1) and Leishmania infantum (n = 3) were identified to species level by restriction fragment length polymorphism analysis. Attempts to sequence PCR products from other pathogens were unsuccessful, but coupled epidemiological and molecular data suggest the presence of Anaplasma platys (n = 5), Babesia vogeli (n = 1) and Ehrlichia canis (n = 1) in semen from dogs. Further experimental studies would be needed to confirm the sexual transmission hypothesis for these VBPs and also the possible implications of these findings for canine reproduction.
Assuntos
Vetores de Doenças , Sêmen , Cães , Animais , Brasil/epidemiologia , Vetores Artrópodes , Ehrlichia canis/genéticaRESUMO
Background: Hemoparasitoses are extremely important in the clinical routine because they affect a large number of dogs. In spite ofthe abundance of studies on this topic, hormonal alterations caused by infection with these agents are still poorly known. Therefore,the goal of this work was to assess the serum levels of thyroid hormones of dogs infected with Ehrlichia canis (E. canis) alone, anddogs infected with E. canis and Babesia canis vogeli (B. vogeli) and/or Anaplasma platys (A. platys) before and after treatment withdoxycycline chlorohydrate. This study also aimed at checking for presence of euthyroid sick syndrome (ESS) in these animals.Materials, Methods & Results: The concentrations of the thyroid hormones total triiodothyronine (TT3), total tetraiodothyronine (TT4),free tetraiodothyronine (FT4), and canine thyroid-stimulating hormone (cTSH) were assessed by chemiluminescence in 12 dogs. Nestedpolymerase chain reaction (nPCR) was used to confirm diagnoses. The dogs were divided into 2 groups: G1, which comprised animalsinfected by E. canis alone, and G2, which included animals simultaneously infected by E. canis and B. vogeli and/or A. platys. Theserum concentrations of the thyroid hormones were measured at two time points: before (D1) and after (D2) the 28-day treatment withgeneric doxycycline chlorohydrate (DC) at a dose of 10 mg/kg SID. On D2, another nPCR was carried out to check the efficacy of thetreatment. On D2, in both groups, all dogs became negative for E. canis; however, 8 animals remained infected or were reinfected byother hemoparasites. On D1, 4 dogs in G1 exhibited low TT3 in conjunction with low TT4; one of the dogs had increased TT3 alone,and another dog had an increased TT3 accompanied by decreased TT4. In G2, on D1, one dog exhibited high TT3 accompanied by adecreased concentration of TT4; 2 dogs had decreased TT4; 2 dogs had increased TT3; and one dog had both TT3 and TT4 decreased...(AU)
Assuntos
Animais , Cães , Cães/parasitologia , Síndromes do Eutireóideo Doente/veterinária , Ehrlichiose/veterinária , Doxiciclina/uso terapêutico , Reação em Cadeia da Polimerase/veterináriaRESUMO
Amblyomma sculptum is a tick that has medical and veterinary importance as, in Brazil, it is the main vector of Rickettsia rickettsii, a disease affecting humans. The presence of ticks was observed outside a residence in a peri-urban area of the Atlantic Forest region in Brazil, as well as on two dogs that lived there. Eighteen A. sculptum adults were seen walking on a cemented pillar at the porch of the house and sheltering inside the pillar's crevices; meanwhile on the dogs, only Rhipicephalus sanguineus sensu lato ticks were found. It is hypothesized that as the dogs circulated in the forest regions, they might have carried A. sculptum to the residence. This situation highlights the role of dogs as possible carriers of Brazilian spotted fever (BSF) tick vectors into human habitation. Strategies for the prevention and control of BSF should consider the hypothesis that ticks infected with R. rickettsii can be harbored in human dwellings in peri-urban areas.
Assuntos
Doenças do Cão , Ixodidae , Rhipicephalus sanguineus , Rickettsia , Febre Maculosa das Montanhas Rochosas , Amblyomma , Animais , Brasil , Doenças do Cão/epidemiologia , Cães , Rickettsia rickettsii/genética , Febre Maculosa das Montanhas Rochosas/epidemiologia , Febre Maculosa das Montanhas Rochosas/veterináriaRESUMO
This study aimed to identify members of the Sarcocystidae family in naturally infected wild birds at a rescue center in the state of Minas Gerais, southeastern Brazil. The heart and brain of 44 wild birds were evaluated by bioassay in mice to detect T. gondii, and extracted DNA was used for nested PCR of the 18S ribosomal DNA gene to detect members of the Sarcocystidae family. The positive samples were sequenced, assembled, edited and compared with sequences deposited in GenBank. Toxoplasma gondii was isolated from six (13.6%) out of 44 birds. Toxoplasma gondii DNA was identified in 10/44 (22.7%) of the birds. The amplified sequences exhibited 100% similarity with the DNA of the ME49 strain of T. gondii. Sarcocystis DNA (99% similarity) was identified in 5/44 (11.4%) of the birds. T. gondii and Sarcocystis spp. are common in wild birds in Minas Gerais, Brazil.
Assuntos
Doenças das Aves , Coccidiose , Sarcocystidae , Animais , Bioensaio , Doenças das Aves/epidemiologia , Doenças das Aves/parasitologia , Aves , Brasil , Coccidiose/epidemiologia , DNA de Protozoário , Camundongos , Reação em Cadeia da Polimerase , RNA Ribossômico 18S/genética , Sarcocystidae/genética , Sarcocystis/genética , Toxoplasma/genética , Toxoplasmose Animal/epidemiologiaRESUMO
This study aimed to identify members of the Sarcocystidae family in naturally infected wild birds at a rescue center in the state of Minas Gerais, southeastern Brazil. The heart and brain of 44 wild birds were evaluated by bioassay in mice to detect T. gondii, and extracted DNA was used for nested PCR of the 18S ribosomal DNA gene to detect members of the Sarcocystidae family. The positive samples were sequenced, assembled, edited and compared with sequences deposited in GenBank. Toxoplasma gondii was isolated from six (13.6%) out of 44 birds. Toxoplasma gondii DNA was identified in 10/44 (22.7%) of the birds. The amplified sequences exhibited 100% similarity with the DNA of the ME49 strain of T. gondii. Sarcocystis DNA (99% similarity) was identified in 5/44 (11.4%) of the birds. T. gondii and Sarcocystis spp. are common in wild birds in Minas Gerais, Brazil.(AU)
O objetivo deste estudo foi identificar membros da família Sarcocystidae em aves silvestres de vida livre naturalmente infectadas e resgatadas no estado de Minas Gerais, Brasil. Coração e cérebro de 44 aves silvestres foram avaliados por bioensaio em camundongos para detecção de T. gondii e extração de DNA para Nested-PCR do gene 18S do DNA ribossomal de membros da família Sarcocystidae. As amostras positivas foram sequenciadas, analisadas, editadas e comparadas com sequências depositadas no GenBank. Toxoplasma gondii foi isolado de seis (13,6%) das 44 aves. DNA de T. gondii foi identificado em 10/44 (22,7%) das 44 aves. As sequências amplificadas exibiram 100% de similaridade com o DNA da cepa ME49 de T. gondii. DNA de Sarcocystis (99% de similaridade) foi identificado em 5/44 (11,4%) das 44 aves. T. gondii e Sarcocystis spp. são encontrados, comumente, em aves silvestres no estado de Minas Gerais, Brasil.(AU)
Assuntos
Animais , Aves Domésticas/parasitologia , Sarcocystidae/patogenicidade , Animais Selvagens/parasitologia , Toxoplasma , Reação em Cadeia da PolimeraseRESUMO
Background: Hemoparasitoses are extremely important in the clinical routine because they affect a large number of dogs. In spite ofthe abundance of studies on this topic, hormonal alterations caused by infection with these agents are still poorly known. Therefore,the goal of this work was to assess the serum levels of thyroid hormones of dogs infected with Ehrlichia canis (E. canis) alone, anddogs infected with E. canis and Babesia canis vogeli (B. vogeli) and/or Anaplasma platys (A. platys) before and after treatment withdoxycycline chlorohydrate. This study also aimed at checking for presence of euthyroid sick syndrome (ESS) in these animals.Materials, Methods & Results: The concentrations of the thyroid hormones total triiodothyronine (TT3), total tetraiodothyronine (TT4),free tetraiodothyronine (FT4), and canine thyroid-stimulating hormone (cTSH) were assessed by chemiluminescence in 12 dogs. Nestedpolymerase chain reaction (nPCR) was used to confirm diagnoses. The dogs were divided into 2 groups: G1, which comprised animalsinfected by E. canis alone, and G2, which included animals simultaneously infected by E. canis and B. vogeli and/or A. platys. Theserum concentrations of the thyroid hormones were measured at two time points: before (D1) and after (D2) the 28-day treatment withgeneric doxycycline chlorohydrate (DC) at a dose of 10 mg/kg SID. On D2, another nPCR was carried out to check the efficacy of thetreatment. On D2, in both groups, all dogs became negative for E. canis; however, 8 animals remained infected or were reinfected byother hemoparasites. On D1, 4 dogs in G1 exhibited low TT3 in conjunction with low TT4; one of the dogs had increased TT3 alone,and another dog had an increased TT3 accompanied by decreased TT4. In G2, on D1, one dog exhibited high TT3 accompanied by adecreased concentration of TT4; 2 dogs had decreased TT4; 2 dogs had increased TT3; and one dog had both TT3 and TT4 decreased...
Assuntos
Animais , Cães , Cães/parasitologia , Síndromes do Eutireóideo Doente/veterinária , Doxiciclina/uso terapêutico , Ehrlichiose/veterinária , Reação em Cadeia da Polimerase/veterináriaRESUMO
Vaccination against Anaplasma marginale has been considered an important control strategy for bovine anaplasmosis. Recently, mice immunized with rMSP1 a linked to carbon nanotubes (MWNT) showed significant immune responses, generating a new possibility for use of an inactivated vaccine. The objective of this study was to investigate the cellular and humoral responses in calves immunized with MWNT+rMSP1a , associated with inactivated vaccine of A. marginale produced in vitro, and evaluate the toxic effects of the MWNT on renal and hepatic function. rMSP1a was covalently linked to MWNT. Inactivated vaccine (AmUFMG2) was produced by cultivating A. marginale in IDE8 cells. Twenty-four Holstein calves were divided (four groups) and immunized subcutaneously with PBS and non-carboxylated MWNT (control, G1), AmUFMG2 (G2), MWNT+rMSP1a (G3), and AmUFMG2 with MWNT+rMSP1a (G4). Blood samples were collected for total leukocyte counts, biochemical profiling and evaluation of the cellular and humoral response. Immunization with MWNT+rMSP1a induced increase in the total number of leukocytes, NK cells, in the lymphocyte populations and higher levels of antibodies compared to calves immunized only with AmUFMG2. Furthermore, MWNT did not induce changes in the biochemical profile. These data indicate that MWNT+rMSP1a were able to induce the immune responses more efficiently than AmUFMG2 alone, without generating toxicity.(AU)
Vacinação contra Anaplasma marginale tem sido considerada uma importante estratégia de controle da anaplasmose bovina. Recentemente, camundongos imunizados com rMSP1a funcionalizada à nanotubos de carbono (MWNT) apresentaram resposta imune significante, gerando nova possibilidade para o uso da vacina inativada. O objetivo desse estudo foi investigar a resposta celular e humoral em bezerros imunizados com MWNT+rMSP1a, associado com a vacina inativada de A. marginale produzida in vitro, e avaliar os efeitos tóxicos dos MWNT nas funções hepática e renal. rMSP1 a foi ligada covalentemente aos MWNT. Vacina inativada (AmUFMG2) foi produzida através do cultivo de A. marginale em células IDE8. Vinte e quatro bezerros Holandeses foram divididos (quatro grupos) e imunizados subcutaneamente com: PBS e MWNT não-carboxilados (controle, G1), AmUFMG2 (G2), MWNT+rMSP1 a (G3), e AmUFMG2 com MWNT+rMSP1a (G4). Amostras de sangue foram coletadas para contagem de leucócitos, perfil bioquímico e avaliação da resposta celular e humoral. Imunização com MWNT+rMSP1a induziu aumento dos leucócitos totais, células NK, na população de linfócitos e altos níveis de anticorpos comparado com animais imunizados apenas com AmUFMG2. Além disso, MWNT não induziu alterações no perfil bioquímico. Esses dados indicam que MWNT+rMSP1a foram capazes de induzir eficientemente a resposta imune comparado com AmUFMG2 sozinho, sem gerar toxicidade.(AU)
Assuntos
Animais , Bovinos , Anaplasma marginale , Anaplasmose/imunologia , Proteína 1 de Superfície de Merozoito , Nanotubos de Carbono , Imunidade Humoral , Vacinas de Produtos InativadosRESUMO
Abstract Vaccination against Anaplasma marginale has been considered an important control strategy for bovine anaplasmosis. Recently, mice immunized with rMSP1 a linked to carbon nanotubes (MWNT) showed significant immune responses, generating a new possibility for use of an inactivated vaccine. The objective of this study was to investigate the cellular and humoral responses in calves immunized with MWNT+rMSP1a , associated with inactivated vaccine of A. marginale produced in vitro, and evaluate the toxic effects of the MWNT on renal and hepatic function. rMSP1a was covalently linked to MWNT. Inactivated vaccine (AmUFMG2) was produced by cultivating A. marginale in IDE8 cells. Twenty-four Holstein calves were divided (four groups) and immunized subcutaneously with PBS and non-carboxylated MWNT (control, G1), AmUFMG2 (G2), MWNT+rMSP1a (G3), and AmUFMG2 with MWNT+rMSP1a (G4). Blood samples were collected for total leukocyte counts, biochemical profiling and evaluation of the cellular and humoral response. Immunization with MWNT+rMSP1a induced increase in the total number of leukocytes, NK cells, in the lymphocyte populations and higher levels of antibodies compared to calves immunized only with AmUFMG2. Furthermore, MWNT did not induce changes in the biochemical profile. These data indicate that MWNT+rMSP1a were able to induce the immune responses more efficiently than AmUFMG2 alone, without generating toxicity.
Resumo Vacinação contra Anaplasma marginale tem sido considerada uma importante estratégia de controle da anaplasmose bovina. Recentemente, camundongos imunizados com rMSP1a funcionalizada à nanotubos de carbono (MWNT) apresentaram resposta imune significante, gerando nova possibilidade para o uso da vacina inativada. O objetivo desse estudo foi investigar a resposta celular e humoral em bezerros imunizados com MWNT+rMSP1a, associado com a vacina inativada de A. marginale produzida in vitro, e avaliar os efeitos tóxicos dos MWNT nas funções hepática e renal. rMSP1 a foi ligada covalentemente aos MWNT. Vacina inativada (AmUFMG2) foi produzida através do cultivo de A. marginale em células IDE8. Vinte e quatro bezerros Holandeses foram divididos (quatro grupos) e imunizados subcutaneamente com: PBS e MWNT não-carboxilados (controle, G1), AmUFMG2 (G2), MWNT+rMSP1 a (G3), e AmUFMG2 com MWNT+rMSP1a (G4). Amostras de sangue foram coletadas para contagem de leucócitos, perfil bioquímico e avaliação da resposta celular e humoral. Imunização com MWNT+rMSP1a induziu aumento dos leucócitos totais, células NK, na população de linfócitos e altos níveis de anticorpos comparado com animais imunizados apenas com AmUFMG2. Além disso, MWNT não induziu alterações no perfil bioquímico. Esses dados indicam que MWNT+rMSP1a foram capazes de induzir eficientemente a resposta imune comparado com AmUFMG2 sozinho, sem gerar toxicidade.
Assuntos
Animais , Bovinos , Portadores de Fármacos , Vacinas Bacterianas/imunologia , Doenças dos Bovinos/microbiologia , Doenças dos Bovinos/prevenção & controle , Nanotubos de Carbono , Anaplasma marginale/imunologia , Imunogenicidade da Vacina , Anaplasmose/prevenção & controle , Imunidade Humoral , Imunidade CelularRESUMO
Vaccination against Anaplasma marginale has been considered an important control strategy for bovine anaplasmosis. Recently, mice immunized with rMSP1 a linked to carbon nanotubes (MWNT) showed significant immune responses, generating a new possibility for use of an inactivated vaccine. The objective of this study was to investigate the cellular and humoral responses in calves immunized with MWNT+rMSP1a , associated with inactivated vaccine of A. marginale produced in vitro, and evaluate the toxic effects of the MWNT on renal and hepatic function. rMSP1a was covalently linked to MWNT. Inactivated vaccine (AmUFMG2) was produced by cultivating A. marginale in IDE8 cells. Twenty-four Holstein calves were divided (four groups) and immunized subcutaneously with PBS and non-carboxylated MWNT (control, G1), AmUFMG2 (G2), MWNT+rMSP1a (G3), and AmUFMG2 with MWNT+rMSP1a (G4). Blood samples were collected for total leukocyte counts, biochemical profiling and evaluation of the cellular and humoral response. Immunization with MWNT+rMSP1a induced increase in the total number of leukocytes, NK cells, in the lymphocyte populations and higher levels of antibodies compared to calves immunized only with AmUFMG2. Furthermore, MWNT did not induce changes in the biochemical profile. These data indicate that MWNT+rMSP1a were able to induce the immune responses more efficiently than AmUFMG2 alone, without generating toxicity.
Assuntos
Anaplasma marginale/imunologia , Anaplasmose/prevenção & controle , Vacinas Bacterianas/imunologia , Doenças dos Bovinos/microbiologia , Doenças dos Bovinos/prevenção & controle , Portadores de Fármacos , Imunogenicidade da Vacina , Nanotubos de Carbono , Animais , Bovinos , Imunidade Celular , Imunidade HumoralRESUMO
In Brazil, Trypanosoma vivax is present in several states. The disease is endemic in Pantanal and Minas Gerais. In Paraná there is still no report of the parasite, but due to the state borders with Mato Grosso do Sul, São Paulo, Paraguay and Argentina, it is believed that the protozoan circulates in the region without diagnosis. The objective of this study was to investigate the prevalence of T. vivax in dairy cattle in the western region of Paraná. For this purpose, 600 blood samples and 400 serum samples were collected from dairy cattle, distributed in 60 and 40 properties, respectively. While buffy coat smears were performed on blood samples, serum samples were used in Indirect Immunofluorescence Reaction. All samples, in both techniques, presented negative results for T. vivax. These results indicate that the studied hemoprotozoan is not circulating among the cattle in the western region of Paraná. However, future work evolving beef cattle must be carried out and preventive measures should be adopted in order to avoid the entry of the parasite in the State of Paraná.(AU)
No Brasil o Trypanosoma vivax está presente em diversos Estados. No Pantanal e em Minas Gerais a doença é endêmica. No Paraná ainda não há relato do parasito, porém como o Estado possui fronteiras com Mato Grosso do Sul, São Paulo, Paraguai e Argentina acredita-se que o protozoário circula na região sem diagnóstico. O objetivo desse trabalho foi pesquisar a prevalência do T. vivax em bovinos leiteiros da região Oeste do Paraná, sendo esse o primeiro estudo com o protozoário no Estado. Para isso, foram coletadas 600 amostras de sangue total e 400 amostras de soro sanguíneo de bovinos leiteiros, distribuídos em 60 e 40 propriedades, respectivamente. Foi realizado esfregaço de papa leucocitária das amostras de sangue e Reação de Imunofluorescência Indireta das amostras de soro. Todas as amostras, em ambas as técnicas, apresentaram resultado negativo para o T. vivax. Esses resultados indicam que o hemoprotozoário pesquisado não está circulando entre os bovinos de leite da região Oeste do Paraná. Porém, novos trabalhos com bovinos de corte devem ser realizados e medidas preventivas devem ser adotadas para evitar a entrada do parasito no Estado do Paraná.(AU)
Assuntos
Animais , Bovinos , Trypanosoma vivax , Tripanossomíase/diagnóstico , Tripanossomíase/prevenção & controle , Doenças Endêmicas/prevenção & controle , BrasilRESUMO
In Brazil, Trypanosoma vivax is present in several states. The disease is endemic in Pantanal and Minas Gerais. In Paraná there is still no report of the parasite, but due to the state borders with Mato Grosso do Sul, São Paulo, Paraguay and Argentina, it is believed that the protozoan circulates in the region without diagnosis. The objective of this study was to investigate the prevalence of T. vivax in dairy cattle in the western region of Paraná. For this purpose, 600 blood samples and 400 serum samples were collected from dairy cattle, distributed in 60 and 40 properties, respectively. While buffy coat smears were performed on blood samples, serum samples were used in Indirect Immunofluorescence Reaction. All samples, in both techniques, presented negative results for T. vivax. These results indicate that the studied hemoprotozoan is not circulating among the cattle in the western region of Paraná. However, future work evolving beef cattle must be carried out and preventive measures should be adopted in order to avoid the entry of the parasite in the State of Paraná.
No Brasil o Trypanosoma vivax está presente em diversos Estados. No Pantanal e em Minas Gerais a doença é endêmica. No Paraná ainda não há relato do parasito, porém como o Estado possui fronteiras com Mato Grosso do Sul, São Paulo, Paraguai e Argentina acredita-se que o protozoário circula na região sem diagnóstico. O objetivo desse trabalho foi pesquisar a prevalência do T. vivax em bovinos leiteiros da região Oeste do Paraná, sendo esse o primeiro estudo com o protozoário no Estado. Para isso, foram coletadas 600 amostras de sangue total e 400 amostras de soro sanguíneo de bovinos leiteiros, distribuídos em 60 e 40 propriedades, respectivamente. Foi realizado esfregaço de papa leucocitária das amostras de sangue e Reação de Imunofluorescência Indireta das amostras de soro. Todas as amostras, em ambas as técnicas, apresentaram resultado negativo para o T. vivax. Esses resultados indicam que o hemoprotozoário pesquisado não está circulando entre os bovinos de leite da região Oeste do Paraná. Porém, novos trabalhos com bovinos de corte devem ser realizados e medidas preventivas devem ser adotadas para evitar a entrada do parasito no Estado do Paraná.
Assuntos
Animais , Bovinos , Doenças Endêmicas/prevenção & controle , Tripanossomíase/diagnóstico , Tripanossomíase/prevenção & controle , Trypanosoma vivax , BrasilRESUMO
Equine granulocytic anaplasmosis is caused by Anaplasma phagocytophilum, a gram negative, obligatory intracellular bacterium, member of Anaplasmataceae family, included in the Rickettsiales order. Little is known about the disease, transmission dynamics, genetic diversity and prevalence in Minas Gerais state, Brazil. This work aimed to do a serosurvey using indirect immunofluorescent assay (IFA) test and evaluation of buffy coat smears, and nested polymerase chain reaction (PCR) as diagnostic methods, to determine the disease situation in horses from two manga-larga marchador breeding farms located in the municipalities of Ataléia e São Vicente de Minas, in Minas Gerais state. It was found that 76% (131/172) of the animals were considered reactive for IFA test, and the total of 12.8% was positive at buffy coat smears analysis. At PCR analysis, it was found 1.94% of the samples positive to the infection. Those samples were sequenced and showed 96% of similarity to A. phagocytophilum from a Ixodes ricinus tick. There is a high frequency of animals with the evidence of contact to A. phagocytophilum on the two evaluated properties in this study, which was proved by positiveness in PCR analysis. New researches must be carried out to better understand the epidemiologic and clinical dynamic of the disease in the state of Minas Gerais.(AU)
A anaplasmose granulocítica equina é causada por uma bactéria gram-negativa, intracelular obrigatória, membro da família Anaplasmataceae, incluída na ordem Rickettsiales e denominada de Anaplasma phagocytophilum. Pouco se sabe sobre a doença, sua dinâmica de transmissão, diversidade genética e prevalência em Minas Gerais, Brasil. Este trabalho teve por objetivo realizar o levantamento sorológico utilizando a reação de imunofluorescência indireta, avaliação direta de capa leucocitária e nested reação em cadeia da polimerase (PCR) como métodos diagnósticos, a fim de avaliar a situação da doença em dois haras de criação de cavalos manga-larga marchador localizados nas cidades de Ataléia e São Vicente de Minas, no estado de Minas Gerais. Foi encontrada prevalência de 76% (131/172) de animais reativos para a reação de imunofluorescência indireta, quando todos os animais das duas propriedades e das duas coletas foram agrupados, e 12,8% dos animais foram positivos na avaliação da capa leucocitária. A reação de imunofluorescência indireta detectou 1,94% das amostras como positivas para o agente. Essas amostras foram submetidas ao sequenciamento de nucleotídeos, e foi observada similaridade de 96% com A. phagocytophilum proveniente de carrapatos Ixodes ricinus. Existe alta prevalência de animais positivos para a infecção por A. phagocytophilum, o que foi provado pela positividade dos animais à PCR. Novas pesquisas devem ser conduzidas a fim de entender a dinâmica epidemiológica e clínica da doença no estado de Minas Gerais.(AU)
Assuntos
Testes Sorológicos/métodos , Reação em Cadeia da Polimerase/métodos , Imunofluorescência/métodos , Cavalos , Anaplasmose , Ixodes , Bactérias Gram-Negativas/patogenicidadeRESUMO
Equine granulocytic anaplasmosis is caused by Anaplasma phagocytophilum, a gram negative, obligatory intracellular bacterium, member of Anaplasmataceae family, included in the Rickettsiales order. Little is known about the disease, transmission dynamics, genetic diversity and prevalence in Minas Gerais state, Brazil. This work aimed to do a serosurvey using indirect immunofluorescent assay (IFA) test and evaluation of buffy coat smears, and nested polymerase chain reaction (PCR) as diagnostic methods, to determine the disease situation in horses from two manga-larga marchador breeding farms located in the municipalities of Ataléia e São Vicente de Minas, in Minas Gerais state. It was found that 76% (131/172) of the animals were considered reactive for IFA test, and the total of 12.8% was positive at buffy coat smears analysis. At PCR analysis, it was found 1.94% of the samples positive to the infection. Those samples were sequenced and showed 96% of similarity to A. phagocytophilum from a Ixodes ricinus tick. There is a high frequency of animals with the evidence of contact to A. phagocytophilum on the two evaluated properties in this study, which was proved by positiveness in PCR analysis. New researches must be carried out to better understand the epidemiologic and clinical dynamic of the disease in the state of Minas Gerais.(AU)
A anaplasmose granulocítica equina é causada por uma bactéria gram-negativa, intracelular obrigatória, membro da família Anaplasmataceae, incluída na ordem Rickettsiales e denominada de Anaplasma phagocytophilum. Pouco se sabe sobre a doença, sua dinâmica de transmissão, diversidade genética e prevalência em Minas Gerais, Brasil. Este trabalho teve por objetivo realizar o levantamento sorológico utilizando a reação de imunofluorescência indireta, avaliação direta de capa leucocitária e nested reação em cadeia da polimerase (PCR) como métodos diagnósticos, a fim de avaliar a situação da doença em dois haras de criação de cavalos manga-larga marchador localizados nas cidades de Ataléia e São Vicente de Minas, no estado de Minas Gerais. Foi encontrada prevalência de 76% (131/172) de animais reativos para a reação de imunofluorescência indireta, quando todos os animais das duas propriedades e das duas coletas foram agrupados, e 12,8% dos animais foram positivos na avaliação da capa leucocitária. A reação de imunofluorescência indireta detectou 1,94% das amostras como positivas para o agente. Essas amostras foram submetidas ao sequenciamento de nucleotídeos, e foi observada similaridade de 96% com A. phagocytophilum proveniente de carrapatos Ixodes ricinus. Existe alta prevalência de animais positivos para a infecção por A. phagocytophilum, o que foi provado pela positividade dos animais à PCR. Novas pesquisas devem ser conduzidas a fim de entender a dinâmica epidemiológica e clínica da doença no estado de Minas Gerais.(AU)
Assuntos
Testes Sorológicos/métodos , Reação em Cadeia da Polimerase/métodos , Imunofluorescência/métodos , Cavalos , Anaplasmose , Ixodes , Bactérias Gram-Negativas/patogenicidadeRESUMO
Visceral leishmaniasis (VL) is a neglected tropical disease with dogs serving as reservoirs for one of its etiological agents, Leishmania infantum. In Brazil, VL control involves culling of seropositive dogs, among other actions. However, the most employed serological tests lack accuracy, and are not able to detect canine visceral leishmaniasis (CVL) during the early stages of infection. Early detection of CVL is highly desirable in order to shorten the contact time between the infected reservoirs and the vectors. In this study, we investigated the ability of two multiepitope proteins, PQ10 and PQ20, to detect CVL at earlier stages than currently employed methods, including ITS-1 conventional PCR. Using serum samples from naturally infected dogs, we observed that ELISA-PQ10 and ELISA-PQ20 were able to detect Leishmania infection at earlier time points as compared with kDNA PCR-RFLP in anti-IgG and anti-IgM assays. Using sera from experimentally infected dogs, we monitored seroconversion using multiepitope proteins, ELISA-crude antigen, as well as ITS-1 conventional and real-time PCR. While seroconversion was detected by ELISA-crude antigen in 16.6% of the dogs, multiepitope proteins were able to detect seroconversion in more than 80% of them. Moreover, the ability of ELISA-PQ10 and ELISA-PQ20 to detect Leishmania infection at earlier time points as compared with conventional PCR was also confirmed in experimental infection dogs' sera. Immunofluorescence to Babesia canis and Ehrlichia canis did not show cross-reactions with ELISA-PQ10/PQ20 positive samples. Results of real-time PCR and ELISA with multiepitope proteins were very similar, with concordances between 80 and 100%. Furthermore, our findings indicated that PQ10 and PQ20 immunoassays can be related to parasite load. ELISA-PQ10 and ELISA-PQ20 are more sensitive diagnostic tools for early CVL detection as compared with other methods They could potentially be used in screening tests due to easy execution and low costs facilities.