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1.
Vet J ; 253: 105387, 2019 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-31685139

RESUMO

Diagnosing canine visceral leishmaniasis (CVL) is difficult because clinical signs of the disease are non-specific and a many infected animals in endemic areas, as in Brazil, are asymptomatic. Serological tests are the most common diagnostic methods employed, but most have limitations. For this reason, the implementation of a rapid, sensitive, and specific diagnostic test for CVL has become increasingly important. In this study, we adapted a chemiluminescent enzyme-linked immunosorbent assay (CL ELISA), using two multi-epitope recombinant proteins (PQ10 and PQ20) and a crude Leishmania antigen produced using promastigotes of L. infantum, as antigens to detect CVL infection in animals from Belo Horizonte. To investigate cross-reactions, samples from dogs with other infections (babesiosis, ehrlichiosis and Trypanosoma cruzi) were tested. Assay performance validations were conducted to analyse parameters such as variability, reproducibility, and stability. CL ELISA sensitivity/specificity with PQ10 antigen was 93.1%/80.0%; with the PQ20 protein 93.1%/96.6%; and with the crude antigen 75%/73.3%. Inter-assay variability and inter-operator coefficient of variation were <7% and <15%, with PQ10 and PQ20, respectively. The accuracy of the CL ELISA was classified as excellent for PQ10 (AUC = 0.95) and PQ20 (AUC = 0.98) and moderate for the crude antigen (AUC = 0.77). The kappa score for qualitative agreement between two plate lots was excellent for PQ10 (0.89) and good for PQ20 (0.65). PQ20 remained more stable than PQ10. The CL ELISA with recombinant proteins is a promising tool to diagnose CVL.


Assuntos
Anticorpos Antiprotozoários/sangue , Antígenos de Protozoários/imunologia , Doenças do Cão/diagnóstico , Leishmania infantum/imunologia , Leishmaniose Visceral/veterinária , Animais , Doenças do Cão/imunologia , Doenças do Cão/parasitologia , Cães , Ensaio de Imunoadsorção Enzimática/veterinária , Feminino , Leishmaniose Visceral/sangue , Leishmaniose Visceral/diagnóstico , Medições Luminescentes/veterinária , Masculino , Sensibilidade e Especificidade , Testes Sorológicos/veterinária
2.
BMC Vet Res ; 15(1): 369, 2019 Oct 25.
Artigo em Inglês | MEDLINE | ID: mdl-31653217

RESUMO

BACKGROUND: Mycoplasma bovis (M. bovis) is an emerging bovine pathogen, leading to significant economic losses in the livestock industry worldwide. Infection can result in a variety of clinical signs, such as arthritis, pneumonia, mastitis and keratoconjunctivitis, none of which are M. bovis-specific. Laboratory diagnosis is therefore important. Serological tests to detect M. bovis antibodies is considered an effective indicator of infection in a herd and often used as a herd test. Combined with clinical judgement, it can also be used to implement control strategies and/or to estimate the disease prevalence within a country. However, due to lack of harmonisation of approaches to testing, and serological tests used by different laboratories, comparisons of prevalence data between countries is often difficult. A network of researchers from six European countries designed and participated in an inter-laboratory trial, with the aim of evaluating the sensitivity (Se) and specificity (Sp) of two commercially available ELISA tests (ID Screen® ELISA (IDvet) and BIO K302 ELISA (BIO-X Diagnostics)) for diagnosis of M. bovis infection. Each laboratory received a blinded panel of bovine sera and tested independently, according to manufacturer's instructions. Western blot analyses (WB) performed by one of the participating laboratories was used as a third diagnostic test in the statistical evaluation of Se and Sp values using latent class analysis. RESULTS: The Se of WB, the ID Screen® ELISA and the BIO K302 ELISA were determined to be 91.8, 93.5 and 49.1% respectively, and corresponding Sp of the three tests were 99.6, 98.6 and 89.6%, respectively. CONCLUSIONS: The present study is, to our knowledge, the first to present an inter-laboratory comparison of the BIO K302 ELISA and the ID Screen® ELISA. Based on our results, the ID Screen® ELISA showed high consistency with WB and performed with higher precision and accuracy than the BIO K302 ELISA.


Assuntos
Doenças dos Bovinos/diagnóstico , Ensaio de Imunoadsorção Enzimática/veterinária , Infecções por Mycoplasma/veterinária , Animais , Western Blotting/veterinária , Bovinos , Doenças dos Bovinos/sangue , Ensaio de Imunoadsorção Enzimática/métodos , Análise de Classes Latentes , Infecções por Mycoplasma/sangue , Infecções por Mycoplasma/diagnóstico , Mycoplasma bovis/imunologia , Sensibilidade e Especificidade , Testes Sorológicos/veterinária
3.
Arq. bras. med. vet. zootec. (Online) ; 71(5): 1649-1658, set.-out. 2019. tab
Artigo em Português | LILACS, VETINDEX | ID: biblio-1038656

RESUMO

Este estudo teve o objetivo de comparar o perfil bioquímico sérico de machos e fêmeas da linhagem pesada de frango de corte, nas idades de quatro, 12 e 20 semanas, em uma unidade de produção industrial, no município de Uberlândia-MG. Após a pesagem das aves, foram coletadas amostras de sangue de 15 aves de cada sexo de cada faixa etária. Os soros obtidos foram avaliados em analisador automático para os seguintes parâmetros bioquímicos: proteína total, albumina, globulinas, ácido úrico, colesterol, triglicérides, gamaglutamiltransferase, aspartato aminotransferase, alanina aminotransferase, creatina quinase, fosfatase alcalina, cálcio e fósforo. Imediatamente após a coleta de cada amostra, avaliou-se a glicemia no sangue total, utilizando-se um glicosímetro. As alterações fisiológicas e metabólicas que as aves apresentaram na fase de recria refletiram na variação dos níveis bioquímicos séricos na maioria dos constituintes avaliados, os quais exibiram diferenças significativas (P<0,05), comparando-se sexo e idade.(AU)


This study aimed to compare the levels of glucose in the blood and serum´s metabolites enzymes and minerals of poultry of heavy lineage of chicken at the age of four, twelve and twenty weeks in an industrial production unit in the city of Uberlândia-MG. After weighing the birds, blood samples were collected from 15 birds of each gender in the three ages. The serum obtained was evaluated in an automatic biochemical analyzer for the following parameters: total protein, albumin, globulin, uric acid, cholesterol, triglycerides, alkaline phosphatase, gamma-glutamyl transferase, aspartate aminotransferase, alanine aminotransferase, creatine kinase, calcium and phosphorus. Immediately after the collection of each sample, we evaluated glucose levels by means of a glycosimeter. The physiological and metabolic changes that birds present in the rearing age reflected in the variation of serum biochemical levels in most constituents evaluated, showing significant differences (P< 0.05) comparing age and gender.(AU)


Assuntos
Animais , Fenômenos Bioquímicos , Testes Sorológicos/veterinária , Galinhas/fisiologia , Galinhas/sangue
4.
PLoS Negl Trop Dis ; 13(9): e0007720, 2019 09.
Artigo em Inglês | MEDLINE | ID: mdl-31513599

RESUMO

Lipophosphoglycan (LPG) is the major surface glycoconjugate of Leishmania protozoan and has an important biological role in host-parasite interactions both in the midgut epithelium of the sand fly vector and in the vertebrate macrophages. Canine leishmaniasis (CanL) is a chronic infectious disease predominantly caused by Leishmania infantum. An early and accurate immunodiagnosis of the disease is crucial for veterinary clinical practice and for disease control. In this work, we evaluated L. infantum LPG as an antigen in an indirect enzyme-linked immunosorbent assay (ELISA) for CanL immunodiagnosis (LPG-ELISA) by testing serum samples from 97 naturally infected dogs with diverse clinical presentations ranging from subclinical infection to severe disease, as evaluated by veterinarian infectologists. Serum samples from healthy dogs from non-endemic areas (n = 68) and from dogs with other infectious diseases (n = 64) were used as controls for assay validation. The performance of the LPG-ELISA was compared with that of an ELISA using the soluble fraction of L. infantum total lysate antigen (TLA). LPG-ELISA presented a superior performance in comparison to TLA-ELISA, with 91.5% sensitivity, 98.5% specificity and 99.7% accuracy. A distinguishing feature of the LPG-ELISA compared to the TLA-ELISA was its higher ability to identify subclinical infection in clinically healthy dogs, in addition to the absence of cross-reactivity with other canine infectious diseases. Finally, LPG-ELISA was compared to TR DPP visceral canine leishmaniasis test, the immunochromatographic test recommended by the Brazilian Ministry of Agriculture. LPG-ELISA exhibited higher values of specificity (98.5% versus 93.1%) and sensitivity (91.5% versus 90.6%) compared to TR DPP. In conclusion, L. infantum-derived LPG was recognized by antibodies elicited during CanL in different infection stages and was shown to be a suitable antigen for specific clinical settings of veterinary diagnosis and for public health usage.


Assuntos
Leishmaniose/veterinária , Animais , Antígenos de Protozoários/análise , Doenças do Cão/diagnóstico , Doenças do Cão/imunologia , Doenças do Cão/parasitologia , Cães , Ensaio de Imunoadsorção Enzimática/métodos , Glicoesfingolipídeos/imunologia , Leishmania infantum/imunologia , Leishmania infantum/isolamento & purificação , Leishmaniose/sangue , Leishmaniose/diagnóstico , Testes Sorológicos/veterinária
5.
Vet Parasitol ; 274: 108920, 2019 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-31493694

RESUMO

Visceral leishmaniasis (VL) is a zoonosis caused by the parasite Leishmania infantum and the dog is its main reservoir in rural and urban areas. The diagnosis of infection is mainly based on the presence of anti-Leishmania IgG antibodies in the serum of infected dogs. In this study, the sensitivity and specificity of qualitative rapid tests (RTs) dual path platform (DPP) Bio-Manguinhos, rapid enzyme-linked immunosorbent assay (ELISA) IDEXX, Kalazar Detect and ALERE, as well as quantitative ELISA Bio-Manguinhos and in-house indirect immunofluorescence assay (IFA) tests were analyzed in sera from infected and uninfected dogs. Serial dilutions of the in-house IFA were compared with RTs and ELISA Bio-Manguinhos. The results showed that none of the tests reached 100% sensitivity and specificity. There was no statistical difference between the analyzed RTs. The most sensitive test was the DPP Bio-Manguinhos (97.9%), while the rapid ELISA IDEXX showed higher specificity (100%). In the treatment setting of infected and/or diseased animals, quantitative tests for monitoring the evolution of antibody titers are required, which indicates the maintenance of in-house IFA in animal handling. Furthermore, we demonstrate that the RTs present higher sensitivity in serum samples with superior antibody titers obtained in the in-house IFA. However, the RTs exhibited false negatives in samples with low titers of antibodies. Among the RTs, only the DPP Bio-Manguinhos presented better performance in this situation. Therefore, the use of RTs for the diagnosis of VL in dogs with low titers of antibodies, such as asymptomatic, should be carefully evaluated.


Assuntos
Doenças do Cão/sangue , Leishmania infantum , Leishmaniose Visceral/veterinária , Testes Sorológicos/veterinária , Animais , Doenças do Cão/diagnóstico , Doenças do Cão/parasitologia , Cães , Leishmaniose Visceral/sangue , Leishmaniose Visceral/diagnóstico , Sensibilidade e Especificidade
6.
Res Vet Sci ; 126: 4-8, 2019 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-31415928

RESUMO

Canine leishmaniosis (CanL) is one of the most important parasitic diseases found in several countries worldwide. Dogs are considered important domestic reservoirs of the parasites, being relevant in the maintenance of transmission cycle of the disease between sandflies and humans. However, the prevalence of asymptomatic infection is considerably higher than that of apparent clinical illness in the infected animals; thus making promptly necessary to diagnose the infection in these animals, which could help to allow to the adoption of more efficient control measures against disease. Parasitological tests, which are considered as gold standard to demonstrate the infection and diagnose the disease, present problems related with their sensitivity. Also, the sample´s collect is considered invasive. As consequence, serological tests could be applied as an additional tool to detect the asymptomatic and symptomatic CanL. For this purpose, distinct recombinant antigens have been studied; however, problems in their sensitivity and/or specificity have been still registered. The present review focus in advances in the identification of new diagnostic targets applied for the CanL diagnose, represented here by recombinant single, combined or chimeric proteins, as well as by peptides that mimic epitopes (mimotopes); which were selected by means of immunoproteomics and phage display.


Assuntos
Doenças do Cão/diagnóstico , Leishmania infantum/imunologia , Leishmaniose/veterinária , Testes Sorológicos/veterinária , Animais , Bacteriófagos , Doenças do Cão/parasitologia , Cães , Epitopos , Humanos , Leishmaniose/diagnóstico , Peptídeos , Proteínas Recombinantes , Sensibilidade e Especificidade
7.
J Vector Borne Dis ; 56(2): 154-158, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31397391

RESUMO

Background & objectives: Cattle population is relatively dense in Nasarawa State (Nigeria) particularly in Keffi and its environs, where there are more Hausa/Fulani settlers whose main occupation is farming and herding. Unfortunately, the area is purportedly described as a "horde of tsetse fly species" which transmits trypanosomes that cause severe disease in humans, livestock and wildlife species. This study was targeted at examining trypanosome species prevalent among cattle breeds reared in Keffi metropolis. Methods: A total of 110 cattle, purely based on availability were screened within five working days for trypanosomes infestation using haematocrit centrifugation technique and buffy coat technique. The breeds of cattle examined included White Fulani (64), Sokoto Gudali (26), N'dama (16) and Muturu (4); reared in Jarmai, Gauta and Keffi North districts of Keffi Local Government Area, Nasarawa State, Nigeria. Data collected were analysed using simple descriptive statistics. Results: It was observed that 18 (16.4%) out of 110 cattle screened were infested with 5 (4.55%) Trypanosoma con- golense and 13 (11.82%) T. vivax. The T. congolense positive cases were 4 (3.64%) in White Fulani and 1(0.91%) in Sokoto Gudali breeds whereas, T. vivax occurrence was 9 (8.18%) in White Fulani breed and 4 (3.64%) in Sokoto Gudali breed. The N'dama and Muturu breeds were absolutely not infested and no mixed infestation was recorded in any of the breeds. Interpretation & conclusion: Trypanosoma vivax and T. congolense are the predominant trypanosome species in the study area affecting mainly Sokoto Gudali and White Fulani breeds. Since, N'dama and Muturu breeds were observed to be trypano-tolerant; intensive breeding strategy, strain upgrading mechanisms and genetic modifications could be adopted to ensure other cattles' survival and prevent disease transmission in the area and beyond.


Assuntos
Testes Sorológicos/veterinária , Trypanosoma/classificação , Tripanossomíase Bovina/diagnóstico , Moscas Tsé-Tsé/parasitologia , Animais , Bovinos , Nigéria/epidemiologia , Trypanosoma vivax , Tripanossomíase Africana , Tripanossomíase Bovina/epidemiologia
8.
Pesqui. vet. bras ; 39(8): 649-654, Aug. 2019. tab
Artigo em Inglês | LILACS, VETINDEX | ID: biblio-1040727

RESUMO

Canine monocytic ehrlichiosis (CME) is an infectious disease caused by the bacterium Ehrlichia canis and transmitted by Rhipicephalus sanguineus sensu lato, a tick with worldwide distribution. When not diagnosed and treated early, disease can be severe. Currently, the disease is confirmed by serological or molecular assays. The objective of this study was to compare a serological assay based on immunochromatography (SPEED® EHRLI immunochromatographic test; BVT, France) and a molecular assay (a screening PCR followed by a nested PCR specific for E. canis) for the diagnosis of E. canis in suspected dogs from Buenos Aires city and southern Greater Buenos Aires, Argentina. Blood samples from 20 clinically healthy dogs (Control Group) and from 80 sick dogs suspected of having CME (Groups 1 to 4) were tested in parallel. Neither the immunochromatographic test nor the PCR assay was able to detect the presence of E. canis in the Control Group. In the group which had been previously tested by serology, the agreement between the tests was low (kappa: 0.200), whereas in the group which had been previously tested by PCR, the concordance between the tests was adequate (kappa: 0.650). The concordance between the tests evaluated in the total population studied was moderate (kappa: 0.496). The results of our study suggest that the use of rapid serological tests as a first approach, together with subsequent confirmation by PCR, will improve the diagnosis of CME.(AU)


A ehrlichiose monocítica canina (CME) é uma doença infecciosa transmitida pelo carrapato Rhipicephalus sanguineus sensu lato com distribuição mundial causada por Ehrlichia canis, que pode produzir uma doença grave se não foi diagnosticada e tratada precocemente. A confirmação da doença é feita diretamente pela detecção do DNA fazendo a reação em cadeia da polimerase (PCR) ou indiretamente por métodos sorológicos. O objetivo deste estudo foi comparar o método sorológico baseado na imunocromatografia e a técnica de PCR para o diagnóstico de E. canis em cães suspeitos da Cidade de Buenos Aires e da região sul da Grande Buenos Aires. As amostras de sangue de 20 cães clinicamente saudáveis (Grupo Controle) e de 80 cães com suspeita clínica de CME (Grupo 1-4) foram avaliadas em paralelo. O diagnóstico serológico foi feito pelo teste imunocromatográfico SPEED® EHRLI (BVT, França). Para a detecção molecular, foi utilizada uma PCR de triagem para amplificar um fragmento de 345 pb do gene que codifica a subunidade 16S do rRNA da família Anaplasmataceae. As amostras positivas depois foram processadas pela PCR aninhada específica para E. canis. No Grupo Controle, a presença de E. canis não foi detectada por PCR ou anticorpos específicos com o teste imunocromatográfico. No grupo em que a sorologia foi solicitada inicialmente (1 e 2), a concordância entre os testes foi baixo (kappa: 0,200) enquanto que no grupo onde o teste inicialmente solicitado foi a PCR, a concordância entre os testes era adequado (kappa: 0,650). A concordância entre os testes avaliados na população total estudada foi moderada (kappa: 0,496). Em conclusão, os resultados do nosso estudo sugerem que o uso de testes serológicos rápidos inicialmente, juntamente com a confirmação subsequente por PCR, permitirá melhorar o diagnóstico de CME.(AU)


Assuntos
Animais , Cães , Ehrlichiose/diagnóstico , Ehrlichiose/veterinária , Ehrlichia canis/isolamento & purificação , Argentina , Testes Sorológicos/veterinária , Reação em Cadeia da Polimerase/veterinária , Cromatografia de Afinidade/veterinária
9.
PLoS Negl Trop Dis ; 13(7): e0007594, 2019 07.
Artigo em Inglês | MEDLINE | ID: mdl-31306417

RESUMO

Though scantly investigated, Leishmania infantum infection and clinical cases of leishmaniasis in cats have been recently reported in several countries of the Mediterranean basin, with large variability in prevalence data. A major limitation in the comparability of the data available is attributed to the differences in diagnostic techniques employed and cat populations sampled. The aim of this study was to assess the prevalence of L. infantum infection in owned cats across Italy by serological and molecular tests and the identification of potential risk factors. Blood samples from 2,659 cats from northern (n = 1,543), central (n = 471) and southern (n = 645) Italy were tested for antibodies against L. infantum, by an immunofluorescence antibody test and for the parasites' DNA, by real-time PCR. Samples were additionally screened for feline leukemia virus (FeLV) and feline immunodeficiency virus (FIV) proviral DNAs. An overall cumulative L. infantum prevalence of 3.9% was recorded by serology (3.3%) and/or qPCR (0.8%), with a higher rate (10.5%) in southern Italy. The risk of L. infantum infection in cats was significantly associated to the geographical areas (South vs North and Centre; p<0.0001), age class (from 19 months to 6 years old vs ≤18 months old, p = 0.0003), neutering status (not neutered vs neutered, p = 0.0028) and FIV infection (p = 0.0051).Though the role of cats in the epidemiology of L. infantum is still debated, our findings indicate that cats are exposed to and/or infected by this protozoan, mainly in endemic regions of Italy. Hence, a standardization of procedures for a prompt diagnosis of L. infantum infection in cats and for screening cat population is crucial for a better understanding of the epidemiology of feline leishmaniasis, and of the potential role of cats in the transmission cycle of zoonotic visceral leishmaniasis.


Assuntos
Doenças do Gato/epidemiologia , Doenças do Gato/parasitologia , Leishmania infantum/isolamento & purificação , Leishmaniose Visceral/epidemiologia , Leishmaniose Visceral/veterinária , Animais , Anticorpos Antiprotozoários/sangue , Gatos , Feminino , Vírus da Imunodeficiência Felina/genética , Vírus da Imunodeficiência Felina/isolamento & purificação , Itália/epidemiologia , Leishmania infantum/genética , Leishmaniose Visceral/diagnóstico , Leishmaniose Visceral/parasitologia , Vírus da Leucemia Felina/genética , Vírus da Leucemia Felina/isolamento & purificação , Masculino , Técnicas de Diagnóstico Molecular/veterinária , Análise Multivariada , Prevalência , Reação em Cadeia da Polimerase em Tempo Real/veterinária , Análise de Regressão , Fatores de Risco , Testes Sorológicos/veterinária , Inquéritos e Questionários
10.
PLoS One ; 14(6): e0219054, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31247024

RESUMO

Schmallenberg virus (SBV), an orthobunyavirus infecting ruminants, emerged in 2011 in Central Europe, spread very rapidly throughout the continent and established an endemic status, thereby representing a constant threat not only to the European livestock population, but also to neighboring countries. Hence, in endemically infected regions, the maintenance and regular verification of diagnostics is needed and in not yet affected regions, suitable diagnostic systems should be established to be prepared for a potential introduction of the disease. In addition, also for the trade of animals into free regions, highly reliable and sensitive diagnostics are of utmost importance. Therefore, a laboratory proficiency trial was initiated to allow for performance evaluations of test systems available for SBV-diagnostics, but also for evaluation of veterinary diagnostic laboratories performing those tests. Ten serum samples (six seropositive, four seronegative) were provided for serological analysis, four of the seropositive samples were provided undiluted, while the remaining samples represented 1/2 and 1/4 dilutions of one of the aforementioned samples in negative serum. Ten further sera (five virus-positive, five negative) were sent to the participants to be analyzed by SBV genome detection methods. A total of 48 diagnostic laboratories from 15 countries of three continents (Europe, Asia, North America) and three kit manufacturers participated in the SBV proficiency test, thereby generating 131 result sets, corresponding to 1310 individual results. The sample panel aimed for serological analysis was tested 72 times; the applied diagnostic methods comprised different commercial ELISAs and standard micro-neutralization tests. The sample set aimed for genome detection was analyzed in 59 approaches by various commercial or in-house (real-time) RT-PCR protocols. Antibody or genome positive samples were correctly identified in every case, independent of the applied diagnostic test system. For seronegative samples, three incorrect, false-positive test results were produced. Virus-negative samples tested false-positive in two cases. Thus, a very high diagnostic accuracy of 99.58% and 99.66% was achieved by the serological and virological methods, respectively. Hence, this ring trial demonstrated that reliable and robust SBV-diagnostics has been established in veterinary diagnostic laboratories in affected and non-affected countries.


Assuntos
Infecções por Bunyaviridae/veterinária , Testes Sorológicos/veterinária , Animais , Anticorpos Antivirais/sangue , Infecções por Bunyaviridae/diagnóstico , Infecções por Bunyaviridae/imunologia , Bovinos , Doenças dos Bovinos/diagnóstico , Doenças dos Bovinos/imunologia , Doenças dos Bovinos/virologia , Doenças Endêmicas/veterinária , Ensaio de Imunoadsorção Enzimática/veterinária , Europa (Continente) , Genoma Viral , Laboratórios , Testes de Neutralização/veterinária , Orthobunyavirus/genética , Orthobunyavirus/imunologia , Reação em Cadeia da Polimerase em Tempo Real/veterinária , Reprodutibilidade dos Testes , Ruminantes , Testes Sorológicos/métodos , Testes Sorológicos/estatística & dados numéricos , Ovinos , Doenças dos Ovinos/diagnóstico , Doenças dos Ovinos/imunologia , Doenças dos Ovinos/virologia , Carneiro Doméstico
11.
Vet Clin Pathol ; 48(2): 305-309, 2019 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-31179564

RESUMO

BACKGROUND: Ehrlichia canis (E canis) infection has been documented in a few small canine case series in Greece. However, there is limited information on the prevalence of exposure to, or the potential risk factors associated with E canis seroreactivity in a large native canine population. OBJECTIVES: The objectives of this study were to evaluate E canis seroprevalence in dogs admitted to a veterinary teaching hospital, and to investigate the potential association between seropositivity and signalment, health status, the serologic assays used, and selected clinical and clinicopathologic abnormalities. METHODS: The medical records of 850 client-owned dogs, tested using three in-office serologic assays, were retrospectively reviewed. RESULTS: The E canis seroprevalence was significantly higher in sick (54.9%) compared with healthy (33.9%) dogs. Seropositivity differed significantly between the serologic assays used in this study (ImmunoComb vs SNAP 3Dx/SNAP 4Dx). Dogs presenting with bleeding tendencies, anemia, thrombocytopenia, leukopenia, pancytopenia, and hyperproteinemia were more likely to be E canis seropositive, and the median hematocrit (HCT), white blood cell (WBC), neutrophil, and platelet values were significantly lower in seropositive compared with seronegative dogs. CONCLUSIONS: A high E canis seroprevalence was documented in a canine population living in an endemic area. Selected clinicopathologic variables might be useful indicators of E canis exposure and could allow the prioritization of serologic testing in the clinical setting.


Assuntos
Doenças do Cão/epidemiologia , Ehrlichia canis/imunologia , Ehrlichiose/veterinária , Animais , Doenças do Cão/microbiologia , Cães , Ehrlichiose/epidemiologia , Ehrlichiose/microbiologia , Feminino , Masculino , Estudos Retrospectivos , Fatores de Risco , Estudos Soroepidemiológicos , Testes Sorológicos/veterinária
12.
PLoS One ; 14(5): e0217290, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31116794

RESUMO

In order to monitor the occurrence of zoonotic agents in pig herds as well as to improve herd health management, the development of new cost-effective diagnostic methods for pigs is necessary. In this study, a protein microarray-based assay for the simultaneous detection of immunoglobulin G (IgG) antibodies against different zoonotic agents and pathogens causing production diseases in pigs was developed. Therefore, antigens of ten different important swine pathogens (Toxoplasma gondii, Yersinia enterocolitica, Salmonella spp., Trichinella spp., Mycobacterium avium, Hepatitis E virus, Mycoplasma hyopneumoniae, Actinobacillus pleuropneumoniae, the porcine reproductive and respiratory syndrome virus, Influenza A virus) were spotted and covalently immobilized as 'antigen-spots' on microarray chips in order to test pig serum for the occurrence of antibodies. Pig serum was sampled at three German abattoirs and ELISA tests for the different pathogens were conducted with the purpose of creating a panel of reference samples for microarray analysis. To evaluate the accuracy of the antigens on the microarray, receiver operating characteristic (ROC) curve analysis using the ELISA test results as reference was performed for the different antigens. High area under curve values were achieved for the antigens of two zoonotic agents: Toxoplasma gondii (0.91), Yersinia enterocolitica (0.97) and for three production diseases: Actinobacillus pleuropneumoniae (0.77), Mycoplasma hyopneumoniae (0.94) and the porcine reproductive and respiratory syndrome virus (0.87). With the help of the newly developed microarray assay, collecting data on the occurrence of antibodies against zoonotic agents and production diseases in pig herds could be minimized to one measurement, resulting in an efficient screening test.


Assuntos
Imunoglobulina G/sangue , Programas de Rastreamento/veterinária , Análise Serial de Proteínas/veterinária , Doenças dos Suínos/diagnóstico , Zoonoses/diagnóstico , Animais , Anticorpos Antibacterianos/sangue , Anticorpos Antiprotozoários/sangue , Anticorpos Antivirais/sangue , Programas de Rastreamento/métodos , Miniaturização , Análise Serial de Proteínas/métodos , Testes Sorológicos/métodos , Testes Sorológicos/veterinária , Sus scrofa/imunologia , Suínos , Doenças dos Suínos/imunologia , Toxoplasma/imunologia , Trichinella/imunologia , Yersinia enterocolitica/imunologia , Zoonoses/imunologia
13.
Trop Anim Health Prod ; 51(7): 1969-1974, 2019 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-31041722

RESUMO

BACKGROUND AND AIM: Burkholderia mallei, the etiologic agent of the disease known as glanders. Clinical and bacteriological diagnosis of glanders is difficult in the early stages of the disease. Currently, mallein (allergic hypersensitivity test) is used for the diagnosis of glanders. The mallein test requires an experienced laboratory person and lasts 48 h. Therefore, in order to quickly diagnose the disease, especially in areas (such as the borders of the country) that cannot be kept animals, new methods should be used to identify the disease. The Rose Bengal is a serological diagnostic test and has been recommended by the World Organization for Animal Health (OIE). In this study, the Rose Bengal test (RBT) was evaluated for the diagnosis of equine glanders, and its diagnostic was compared with mallein test. MATERIALS AND METHODS: Sera from 70 naturally infected culture-positive horses, 3 equines that were sensitized by injecting antigen and 110 healthy equines were tested. Specificity and sensitivity of RBT and mallein test when testing culture-positive equines were calculated. RESULTS: Diagnosis of glanders with both methods yield the same results, but Rose Bengal test is much faster than mallein test for diagnosis of equine glanders. CONCLUSION: By comparative RBT with mallein test, it can be considered, RBT test has been used for rapid detection of glanders with features such as, ease of use and can be applicable without specialized equipment and trained personnel. Because the RBT is simpler and rapid to perform, the inclusion of the test as a supplementary test for the diagnosis of glanders in field conditions is recommended.


Assuntos
Burkholderia mallei/isolamento & purificação , Mormo/diagnóstico , Rosa Bengala/química , Testes Sorológicos/veterinária , Animais , Burkholderia mallei/química , Cavalos , Fatores de Tempo
14.
Parasit Vectors ; 12(1): 228, 2019 May 14.
Artigo em Inglês | MEDLINE | ID: mdl-31088526

RESUMO

BACKGROUND: Histomonosis, caused by the protozoan parasite Histomonas meleagridis, is a severe disease especially in turkeys where it can cause high mortalities. Recently, outbreaks were described in which turkey hens showed no clinical signs despite high mortalities in toms, from which they were separated only by a wire fence. The present study investigated three similar outbreaks of histomonosis whereby in two of them only a few hens were being affected and none in the third. Hens from all flocks were kept until end of production and slaughtered as scheduled. However, in all three cases, the disease progressed in toms reaching nearly 100% within two weeks. METHODS: Following diagnosis of the disease, tissue samples were obtained from toms and hens at necropsy. Environmental dust, cloacal swabs and blood were taken on three successive farm visits within compartments of hens and toms and tested by real-time PCR or ELISA. The DNA from a total of 18 samples positive for H. meleagridis was further subjected to conventional PCR utilizing the 18S rRNA primers and sequenced for phylogenetic analysis. RESULTS: All tissue samples and some cloacal swabs were tested positive. Dust samples confirmed the presence of H. meleagridis DNA that spread within entire houses up to 6 weeks after the first clinical signs of histomonosis. Sequence analysis of the 18S rRNA locus demonstrated the presence of the same strain in birds of both sexes within each of the turkey houses. Investigation of serum samples two weeks post-initial diagnosis and prior to euthanasia resulted in antibody detection in 73% of toms and 70% of hens. Until the end of the investigation the number of positive hens per farm increased up to 100% with mean OD-values approaching those noticed in toms prior to euthanasia. CONCLUSIONS: For the first time it could be demonstrated that turkey hens kept in the same house as toms became infected during fatal outbreaks in toms. This highlights the value of different diagnostics methods in order to trace the parasite in connection with the host response. The strange phenomenon that only single hens succumb to the diseases despite being infected requires further investigations.


Assuntos
Doenças das Aves Domésticas/mortalidade , Infecções Protozoárias em Animais/mortalidade , Trichomonadida/genética , Perus/parasitologia , Animais , Cloaca/parasitologia , Surtos de Doenças , Feminino , Masculino , Reação em Cadeia da Polimerase/veterinária , Doenças das Aves Domésticas/parasitologia , Infecções Protozoárias em Animais/patologia , RNA Ribossômico 18S/genética , Análise de Sequência de DNA , Testes Sorológicos/veterinária , Fatores Sexuais , Trichomonadida/isolamento & purificação
15.
Transbound Emerg Dis ; 66(5): 2175-2179, 2019 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-31144447

RESUMO

The native Eurasian wild boar (Sus scrofa) is a reservoir of Mycobacterium bovis, the causative agent of animal tuberculosis (TB), a chronic disease in livestock, companion animals and wild mammals. Cases of M. bovis infection in wild boar or feral pig have been reported worldwide, making early detection a priority in the eradication of the disease. Point-of-care diagnostic tests, such as low cost lateral flow assays, provide high specificity and sensitivity and can be performed on site, an essential requirement for a rapid screening of wildlife. A lateral flow assay, LFA, (INgezim TB CROM Ab) for the detection of M. bovis-specific antibodies in wild boar serum and blood has been developed based on MPB83, one of the major immunogenic antigens of the bacterium. A total of 140 samples of wild boar serum, well-characterized by Mycobacterium tuberculosis complex culture and TB compatible post-mortem lesions, have been analysed with LFA, and results were compared with one in-house and two commercial Enzyme-linked Immunosorbent Assays (ELISA), INgezim TB Porcine and INgezim Tuberculosis DR. In experimental samples, the achieved values of sensitivity of the different techniques ranged from 84.3% to 92.1% and the specificity was 100% in all of them. In field animals, specificity ranged from 96% to 100%, whereas sensitivity ranged from 48% to 64% in juvenile wild boar, increasing to 93.3%-100% in adult wild boar. In particular, the total sensitivity and specificity values obtained with the new LFA were 83% and 97%, respectively, indicating that INgezim TB CROM Ab could be used as a first approach for the surveillance of TB in wild boar, with a special applicability for animal-side testing.


Assuntos
Testes Diagnósticos de Rotina/veterinária , Mycobacterium bovis/isolamento & purificação , Testes Sorológicos/veterinária , Doenças dos Suínos/diagnóstico , Tuberculose/veterinária , Animais , Animais Selvagens , Testes Diagnósticos de Rotina/métodos , Testes Sorológicos/métodos , Espanha , Suínos , Tuberculose/diagnóstico
16.
Vet Parasitol Reg Stud Reports ; 16: 100278, 2019 04.
Artigo em Inglês | MEDLINE | ID: mdl-31027599

RESUMO

This study was carried out to evaluate the application of CATT/T. evansi, crude and recombinant (TeGM6-4r) antigen ELISAs in the diagnosis of camel trypanosomosis caused by two trypanosome species, T. evansi and T. vivax, in Sudan. Concurrently, the current situation of camel trypanosomosis was investigated based on the results of a serological analysis. The recombinant tandem repeat antigen TeGM6-4r is conserved among salivarian trypanosome species and was highly sensitive in the detection Trypanozoon, and T. vivax. It has been validated in the diagnosis of surra in cattle and water buffalo but not in camels. A comparative evaluation of a crude antigen ELISA and a recombinant antigen GM6 (rTeGM6-4r) ELISA was performed using 189 blood samples, which included 148 samples obtained from different camel herds in Eastern Sudan and 41 samples from camels that had been brought from Western Sudan to local markets. The results showed that the rTeGM6-4r ELISA detected the greatest number of positive samples (n = 118, 62%), while CATT/T. evansi and the crude antigen ELISA detected the lowest number of positive samples (n = 73, 39%). The kappa value of rTeGM6-4r as compared to TeCA ELISA was 0.5515, which indicated moderate agreement. We concluded that the rTeGM6-4r ELISA is the test of choice for use in screening camel for trypanosomosis caused by T. evansi and T. vivax in Sudan.


Assuntos
Anticorpos Antiprotozoários/sangue , Camelus/parasitologia , Ensaio de Imunoadsorção Enzimática/veterinária , Trypanosoma/imunologia , Tripanossomíase Africana/veterinária , Testes de Aglutinação/veterinária , Animais , Proteínas Recombinantes/imunologia , Estudos Soroepidemiológicos , Testes Sorológicos/veterinária , Sudão/epidemiologia , Trypanosoma/classificação , Trypanosoma vivax/imunologia , Tripanossomíase Africana/diagnóstico , Tripanossomíase Africana/epidemiologia , Tripanossomíase Africana/imunologia , Glicoproteínas Variantes de Superfície de Trypanosoma/imunologia
17.
J Vet Intern Med ; 33(3): 1434-1439, 2019 May.
Artigo em Inglês | MEDLINE | ID: mdl-31004383

RESUMO

Gastrointestinal (GI) pythiosis is a severe and often fatal disease in dogs that traditionally has been poorly responsive to medical treatment. Although aggressive surgical resection with wide margins is the most consistently effective treatment, lesion location and extent often preclude complete resection. Recently, it has been suggested that the addition of anti-inflammatory doses of corticosteroids may improve outcome in dogs with nonresectable GI pythiosis. This report describes 3 dogs with colonic pythiosis in which complete resolution of clinical signs, regression of colonic masses, and progressive decreases in serological titers were observed after treatment with itraconazole, terbinafine, and corticosteroids. This treatment protocol represents a promising treatment for dogs with GI pythiosis in which surgical intervention is not feasible.


Assuntos
Doenças do Cão/tratamento farmacológico , Itraconazol/uso terapêutico , Prednisona/uso terapêutico , Pitiose/tratamento farmacológico , Terbinafina/uso terapêutico , Animais , Anti-Inflamatórios/administração & dosagem , Anti-Inflamatórios/uso terapêutico , Antifúngicos/administração & dosagem , Antifúngicos/uso terapêutico , Colo/patologia , Doenças do Cão/microbiologia , Cães , Gastroenteropatias/tratamento farmacológico , Gastroenteropatias/microbiologia , Gastroenteropatias/veterinária , Itraconazol/administração & dosagem , Prednisona/administração & dosagem , Pythium/imunologia , Testes Sorológicos/veterinária , Terbinafina/administração & dosagem
18.
Vector Borne Zoonotic Dis ; 19(4): 290-294, 2019 04.
Artigo em Inglês | MEDLINE | ID: mdl-30932773

RESUMO

INTRODUCTION: Hantaviruses are a group of globally distributed rodent-associated viruses, some of which are responsible for human morbidity and mortality. Sin Nombre orthohantavirus, a particularly virulent species of hantavirus associated with Peromyscus spp. mice, is actively monitored by the Department of Public Health in California (CDPH). Recently, CDPH documented high (40%) seroprevalence in a potentially novel reservoir species, the cactus mouse (Peromyscus eremicus) in Death Valley National Park. METHODS: This study was performed in the extremely isolated Mojave Desert Amargosa River valley region of southeastern Inyo County, California, 105 km from Death Valley, approximately over the same time interval as the CDPH work in Death Valley (between 2011 and 2016). Similar rodent species were captured as in Death Valley and were tested for select hantaviruses using serology and RT-PCR to assess risk to human health and the conservation of the endemic endangered Amargosa vole. RESULTS: Among 192 rodents tested, including 56 Peromyscus spp., only one seropositive harvest mouse (Reithrodontomys megalotis) was detected. DISCUSSION: These data highlight the heterogeneity in the prevalence of hantavirus infection even among nearby desert communities and suggest that further studies of hantavirus persistence in desert environments are needed to more accurately inform the risks to public health and wildlife conservation.


Assuntos
Anticorpos Antivirais/sangue , Roedores/sangue , Roedores/virologia , Vírus Sin Nombre/imunologia , Animais , California , Reservatórios de Doenças/virologia , Testes Sorológicos/veterinária
19.
Transbound Emerg Dis ; 66(4): 1674-1692, 2019 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-30980699

RESUMO

Brucella-exposure and infection is increasingly recognized in marine mammals worldwide. To better understand the epidemiology and health impacts of Brucella spp. in marine mammals of Brazil, molecular (conventional PCR and/or real-time PCR), serological (Rose Bengal Test [RBT], Competitive [c]ELISA, Serum Agglutination Test [SAT]), pathological, immunohistochemical (IHC) and/or microbiological investigations were conducted in samples of 129 stranded or by-caught marine mammals (orders Cetartiodactyla [n = 124], Carnivora [n = 4] and Sirenia [n = 1]). Previous serological tests performed on available sera of 27 of the 129 animals (26 cetaceans and one manatee), indicated 10 seropositive cetaceans. Conventional PCR and/or real-time PCR performed in cases with available organs (n = 119) and/or blood or swabs (n = 10) revealed 4/129 (3.1%) Brucella-infected cetaceans (one of them with positive serology; the remaining three with no available sera). Pathological, IHC and/or microbiological analyses conducted in PCR/real-time PCR and/or seropositive cases (n = 13) revealed Brucella-type lesions, including meningitis/meningoencephalitis, pneumonia, necrotizing hepatitis, pericarditis and osteoarthritis in some of those animals, and positive IHC was found in all of them (excepting two live-stranded animals without available organs). Brucella spp. culture attempts were unsuccessful. Our results demonstrated exposure, asymptomatic, acute and chronic Brucella sp. infection in several cetacean species in the Brazilian coast, highlighting the role of this pathogen in stranding and/or death, particularly in Clymene dolphin (Stenella clymene) and short-finned pilot whale (Globicephala macrorhynchus) off Ceará State. Novel hosts susceptible to Brucella included the franciscana (Pontoporia blainvillei), the Guiana dolphin (Sotalia guianensis) and the spinner dolphin (Stenella longirostris). Additionally, three coinfection cases involving Brucella spp. and cetacean morbillivirus, Edwarsiella tarda and Proteus mirabilis were detected. To the best of our knowledge, this is the first long-term and large-scale survey of Brucella spp. in marine mammals of South America, widening the spectrum of susceptible hosts and geographical distribution range of this agent with zoonotic potential.


Assuntos
Brucella/fisiologia , Brucelose/veterinária , Cetáceos , Otárias , Sirênios , Animais , Brasil/epidemiologia , Brucelose/epidemiologia , Brucelose/microbiologia , Brucelose/patologia , Ensaio de Imunoadsorção Enzimática/veterinária , Reação em Cadeia da Polimerase/veterinária , Testes Sorológicos/veterinária
20.
Vet Clin North Am Small Anim Pract ; 49(4): 703-718, 2019 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-30975506

RESUMO

Vector-borne disease and idiopathic immune-mediated disease present similarly. Diagnostic panels that include multiple organisms help detect infection and identify coinfections. Comprehensive diagnostic panels that combine polymerase chain reaction (PCR) and serology should be used in initial screening to maximize sensitivity and identify infection. Repeat testing using PCR is warranted in dogs at high risk of infection with organisms that circulate in blood in low numbers or intermittently. Convalescent serologic testing can help diagnose acute infection. This article discusses the pathophysiology and epidemiology of the organisms, panel selection, and how to recognize when more aggressive testing for an organism is warranted.


Assuntos
Vetores Artrópodes , Doenças do Cão/diagnóstico , Reação em Cadeia da Polimerase/veterinária , Testes Sorológicos/veterinária , Animais , Doenças do Cão/sangue , Doenças do Cão/transmissão , Cães
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