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1.
Exp Parasitol ; 206: 107771, 2019 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-31585116

RESUMO

A PCR targeting mitochondrial cytochrome oxidase subunit III (cox3) for molecular detection of Babesia gibsoni infection in dogs has been developed in this study. Fifty blood samples from suspected clinical cases from dogs, brought to the veterinary college clinics, were examined for presence of B. gibsoni using conventional diagnosis by microscopic examination of Giemsa stained thin blood smears. In addition, species specific PCRs targeting ITS-1 region (BgITS-1 PCR) and nested PCR targeting 18S ribosomal RNA gene (Bg18SnPCR) were carried out. A 634 bp PCR fragment of B. gibsoni cox3 gene was amplified in positive samples from three geographical locations of Satara, Wai and Pune in Maharashtra state of India. From analysis of the sequence of the B. gibsoni cox3 gene, we found that the Indian isolate had 96-98% similarity to the isolate from Japan and China. Post sequencing, de-novo diagnostic primer pair for species specific amplification of 164 bp fragment of B. gibsonicox3 was designed and the PCR was standardized. The diagnostic results of de-novo Bgcox3 PCR were compared with BgITS-1 PCR and Bg18S nPCR. Thin blood smears detected 22% (11/50) samples positive for small form of Babesia species. The BgITS-1 PCR detected 25% samples (15/50) as positive and Bg18S nPCR detected 80% (40/50) B. gibsoni positive samples. The de-novo Bgcox3 PCR detected 66% (33/50) samples positive for B. gibsoni (at 95% CI). The analytical sensitivity of cox3 PCR was evaluated as 0.000003% parasitaemia or 09 parasites in 100  µl of blood. The de-novo diagnostic cox3 PCR did not cross react with control positive DNA from other haemoprotozoa and rickettsia like B. vogeli, Hepatozoon canis, Trypanosoma evansi, Ehrlichia canis and Anaplasma platys. Statistically, cox3 PCR had better diagnostic efficiency than ITS-1 PCR in terms of sensitivity (p = 0.0006). No statistically significant difference between results of cox3 PCR and 18S nPCR was observed (p = 0.1760). Kappa values estimated for each test pair showed fair to moderate agreement between the observations. Specificity of Bgcox3 PCR was 100% when compared with microscopy or BgITS-1 PCR. Sensitivity of Bgcox3 PCR was 100% when compared with that of Bg18S nPCR.


Assuntos
Babesia/isolamento & purificação , Babesiose/diagnóstico , Doenças do Cão/diagnóstico , Complexo IV da Cadeia de Transporte de Elétrons/genética , Mitocôndrias/enzimologia , Animais , Babesia/classificação , Babesia/genética , Babesiose/parasitologia , Sequência de Bases , Reações Cruzadas , DNA Espaçador Ribossômico/química , Doenças do Cão/parasitologia , Cães , Eritrócitos/parasitologia , Funções Verossimilhança , Filogenia , Reação em Cadeia da Polimerase/veterinária , Valor Preditivo dos Testes , RNA Ribossômico 18S/análise , Sensibilidade e Especificidade , Alinhamento de Sequência/veterinária
2.
Vet Parasitol ; 273: 17-23, 2019 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-31442888

RESUMO

The protozoan parasite Tritrichomonas foetus may cause severe diarrhea in cats all over the world. In order to evaluate the methodology in coprological molecular diagnosis of feline tritrichomonosis, we compared previously published ("old") and newly developed ("novel") loop-mediated isothermal amplification (LAMP) (targeted to the T. foetus ß-tubulin and the elf1α 1 gene, respectively) as well as an old conventional and an old and novel real-time PCR (all targeted to overlapping regions of T. foetus rDNA) assays regarding their diagnostic sensitivities and specificities. Here, the novel real-time PCR yielded the best methodical performance in that a sensitivity with a detection limit of <0.1 trophozoites (corresponding to ca.<0.13 trophozoites per mg feces) and a maximal specificity for diagnosis of Tritrichomonas spp. was achieved. The other test systems exhibited either an approximately 10-times lower sensitivity (<1 trophozoite corresponding to ca.<1.3 trophozoites per mg feces) (conventional PCR and both LAMP assays) or a lower specificity (old real-time PCR). Conversely, the diagnostic performance assessed with clinical fecal samples from cats demonstrated identical sensitivities (8 of 20 samples tested were positive) for the novel PCR and both LAMP assays. Diagnostic sensitivities were significantly higher than those found for the old real-time (5 positive samples) and conventional PCR (6 positive samples), respectively. Accordingly, our data suggested the novel PCR and both LAMP assays to be well suited molecular tools for direct (i.e. without including an in vitro cultivation step) coprological diagnosis of tritrichomonosis in cats. Interestingly, relative high (novel LAMP, 7 positive samples) to at least moderate (old LAMP, 6 positive samples and 1 sample with equivocal score) diagnostic sensitivities were also achieved by testing clinical samples upon simple visual inspection of colorimetric changes during the LAMP amplification reactions. Accordingly, both LAMP assays may serve as practical molecular tools to perform epidemiological studies on feline (and bovine as well as porcine) tritrichomonosis under simple laboratory conditions.


Assuntos
Doenças do Gato/diagnóstico , Técnicas de Amplificação de Ácido Nucleico/veterinária , Reação em Cadeia da Polimerase/veterinária , Infecções Protozoárias em Animais/diagnóstico , Tritrichomonas foetus , Animais , Doenças do Gato/parasitologia , Gatos , Fezes/parasitologia , Limite de Detecção , Técnicas de Amplificação de Ácido Nucleico/normas , Reação em Cadeia da Polimerase/normas , Sensibilidade e Especificidade
3.
Rev Bras Parasitol Vet ; 28(3): 489-492, 2019 Aug 12.
Artigo em Inglês | MEDLINE | ID: mdl-31411313

RESUMO

Cryptosporidium is a protozoan parasite with a wide range of hosts, including humans. However, only a few Cryptosporidium species have been described in birds (C. meleagridis, C. baileyi, C. galli and C. avium). The aim of this study was to investigate the occurrence of Cryptosporidium spp. in feces of eared doves (Zenaida auriculata), followed by molecular characterization of the parasite. A total of 196 animals of both sexes were trap-captured; the animals were culled and the intestinal contents were collected for DNA extraction. After extraction, a nested-PCR (nPCR), which amplifies a fragment of the 18S rRNA gene of Cryptosporidium spp., was performed. The amplicons obtained were purified and sequenced. PCR analysis revealed that 30 animals (15.3%) were positive for Cryptosporidium spp. There was no significant sex-dependent enrichment of Cryptosporidium occurrence (p > 0.05). Only 15 out of the 30 positive samples were successfully sequenced and their species determined, of which, 13 (86.7%) and 2 (13.3%) were C. meleagridis and C. galli, respectively. Herein, we present for the first time a molecular characterization of Cryptosporidium from feces of eared doves (Z. auriculata) and propose that these birds are a potential source of C. meleagridis infection in humans.


Assuntos
Doenças das Aves/parasitologia , Columbidae/parasitologia , Cryptosporidium/isolamento & purificação , Animais , DNA de Protozoário/genética , Fezes/parasitologia , Feminino , Filogenia , Reação em Cadeia da Polimerase/veterinária , RNA Ribossômico 18S/genética
4.
Exp Parasitol ; 205: 107714, 2019 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-31279927

RESUMO

The objective of the present study was to evaluate the clinical signs, electrocardiographic signs and evolution of histopathological lesions in the heart of sheep experimentally infected by Trypanosoma vivax during the acute and chronic phases of infection as well as to investigate the presence of parasitic DNA in the heart using polymerase chain reaction (PCR). Twenty-two male sheep were divided into the following four groups: G1, which consisted of six sheep infected by T. vivax that were evaluated until 20 days post-infection (dpi; acute phase); G2, which consisted of six sheep infected by T. vivax that were evaluated until 90 dpi (chronic phase); and G3 and G4 groups, which each consisted of five uninfected sheep. At the end of the experimental period, electrocardiographic evaluations and necroscopic examinations were performed. Fragments of the heart were collected and stained by Hematoxylin-Eosin and Masson's trichrome, and the fragments were also evaluated by PCR for T. vivax. G2 animals presented clinical signs suggestive of heart failure and electrocardiogram alterations characterized by prolonged P, T and QRS complex durations as well as by a cardiac electrical axis shift to the left and increased heart rate. In these animals, mononuclear multifocal myocarditis and interstitial fibrosis were also observed. PCR revealed positivity for T. vivax in two G1 animals and in all G2 animals. Thus, these findings suggested that T. vivax is responsible for the occurrence of cardiac lesions, which are related to heart failure, electrocardiographic alterations and mortality of the infected animals.


Assuntos
DNA de Protozoário/isolamento & purificação , Insuficiência Cardíaca/veterinária , Coração/parasitologia , Doenças dos Ovinos/parasitologia , Trypanosoma vivax/patogenicidade , Tripanossomíase Africana/veterinária , Doença Aguda , Animais , Anticorpos Antiprotozoários/sangue , Doença Crônica/veterinária , Eletrocardiografia/veterinária , Técnica Indireta de Fluorescência para Anticorpo/veterinária , Insuficiência Cardíaca/mortalidade , Insuficiência Cardíaca/parasitologia , Imunoglobulina G/sangue , Masculino , Miocárdio/patologia , Parasitemia/veterinária , Pericardite/parasitologia , Pericardite/patologia , Pericardite/veterinária , Reação em Cadeia da Polimerase/veterinária , Distribuição Aleatória , Ovinos , Doenças dos Ovinos/mortalidade , Doenças dos Ovinos/patologia , Trypanosoma vivax/genética , Trypanosoma vivax/imunologia , Trypanosoma vivax/isolamento & purificação , Tripanossomíase Africana/complicações , Tripanossomíase Africana/mortalidade , Tripanossomíase Africana/patologia
5.
Vet Parasitol ; 271: 1-6, 2019 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-31303197

RESUMO

Heterakis gallinarum is a widely distributed cecal nematode that parasitizes gallinaceous birds including chickens and turkeys. H. gallinarum infection poses a problem for the poultry industry as the nematode egg serves as a vector for the protozoan parasite, Histomonas meleagridis, the causative agent of histomonosis. The only means of detecting H. gallinarum in the environment is microscopic identification of the eggs in soil or feces; however, H. gallinarum eggs are often mistaken for those of Ascaridia galli. Three primer sets were designed from sequences cloned from the H. gallinarum genome to develop a diagnostic PCR. Each of these primer sets amplified a single product from H. gallinarum, but were unable to amplify DNA from H. meleagridis, Ascaridia galli, or Cestode sp. H. gallinarum DNA was amplified from Lumbricus sp. (earthworms) and Alphitobius diaperinus (darkling beetles), confirming that the earthworm acts as a paratenic host for H. gallinarum and suggesting that the darkling beetle may be a carrier for this nematode.


Assuntos
Infecções por Ascaridida/veterinária , Ascaridídios/genética , Monitoramento Ambiental/métodos , Reação em Cadeia da Polimerase/veterinária , Doenças das Aves Domésticas/diagnóstico , Doenças das Aves Domésticas/parasitologia , Animais , Ascaridídios/parasitologia , Infecções por Ascaridida/diagnóstico , Besouros/parasitologia , DNA de Helmintos/genética , Oligoquetos/parasitologia , Doenças das Aves Domésticas/transmissão , Infecções Protozoárias em Animais/transmissão , Solo/parasitologia , Trichomonadida/fisiologia
6.
Prev Vet Med ; 169: 104696, 2019 Aug 01.
Artigo em Inglês | MEDLINE | ID: mdl-31311632

RESUMO

Canine babesiosis is a serious disease among tick-borne haemoprotozoan diseases, globally. The present study was envisaged for carrying out thorough investigation of the disease among working dogs of organised kennels situated in different agro-climatic zones of India as comprehensive understanding of the disease from this country was pertinently lacking. During the study period of three years (2012-2014), 330 dogs suspected for babesiosis were examined for clinicopathology by their physical examination, haematological and biochemical parameters estimation, while the detection of apicomplexan parasites was confirmed by using various diagnostic techniques i.e. by conventional microscopy, by two different Babesia specific 18S rRNA based PCR protocols (conventional/simple PCR and nested PCR assays) followed by sequencing of obtained PCR amplicons for Babsesia spp. identification. Out of 330 clinical cases screened 5.15% (17/330), 9.09% (30/330) and 15.45% (51/330) were found to be positive in microscopic examination, simple- and nested- PCR assay, respectively. Comparative statistical analyses of these diagnostic assay results revealed that significant difference exists among the three diagnostic methodologies and thus it is recommended that the nested PCR technique be relied upon as a screening molecular assay and also for epidemiological studies of the disease in this country. Phylogenetic analysis based on 18S rRNA depicted the monophyletic nature and clonal expansion among all the B. gibsoni, under study. Sequencing results of PCR amplicons revealed that B. gibsoni has predominantly established itself over B. vogeli as former was incriminated in 47 cases while latter was confirmed in only four animals. Based on the clinical severity, these 51 affected animals were classified into three main groups' of 17 animals each viz., apparently healthy-, simple or uncomplicated babesiosis- and atypical or complicated babesiosis- group. Haematological and biochemical profiling of these dogs confirmed the characteristics findings of infection by both the Babesia spp. It was observed that the infection by small form of Babesia (B. gibsoni) is posing a significant therapeutic challenge and chemosterilization by commonly prescribed anti-protozoal drugs was not achieved as clinical relapses were often observed. The clinical signs, sequence based confirmation and severity of the infection suggested that there is a positive selection of B. gibsoni (smaller form) over B. vogeli (larger form) in this country and raises serious concerns as prognosis in former is considered to be poor compared to latter. Thus, these findings have opened new paradigms for planning of pragmatic control strategies against this emerging canine health problem.


Assuntos
Babesia/genética , Babesiose/epidemiologia , Babesiose/genética , Doenças do Cão/epidemiologia , Doenças do Cão/genética , Animais , Babesia/isolamento & purificação , Babesiose/sangue , Babesiose/patologia , Doenças do Cão/sangue , Doenças do Cão/parasitologia , Cães , Feminino , Abrigo para Animais , Índia/epidemiologia , Masculino , Epidemiologia Molecular , Técnicas de Amplificação de Ácido Nucleico/veterinária , Reação em Cadeia da Polimerase/veterinária , RNA Ribossômico 18S/genética , Análise de Sequência de DNA
7.
Prev Vet Med ; 169: 104708, 2019 Aug 01.
Artigo em Inglês | MEDLINE | ID: mdl-31311635

RESUMO

Brucella spp. commonly infect humans in various regions worldwide. Human brucellosis mainly spreads through the consumption of contaminated raw dairy products and meat from domestic livestock (water buffalo, goats, sheep, cattle, pigs and camels). In this regard, the origin and routes of transmission of this bacterium should be carefully determined in order to control the source of infection. This study aimed to evaluate the rate of Brucella spp. contamination of camel milk samples sent for analysis to the national brucellosis laboratory during 2018 in Iran. For this purpose, 96 milk samples from 96 dairy camel herds were randomly collected from two provinces and investigated for the presence of Brucella spp contaminations by both bacterial culture method and polymerase chain reaction (PCR). No clinical manifestation of brucellosis was reported in camels from which milk samples were collected. Using the culture method, three milk samples (3%) originating from two camels of Isfahan province (4%) and one camel from the Semnan province (2%), were contaminated with Brucella abortus. According to PCR analyses, B. abortus gene was detected in 14 (14.5%) milk samples, including 9 and 5 samples from Isfahan (18%) and Semnan (11%) province, respectively. PCR method revealed significant differences (p = 0.02) in the level of contamination with B. abortus between milk samples collected from two regions. These results represent the first report regarding the isolation of B. abortus from raw camel milk in Iran and highlight the importance to screen apparent healthy camels. Therefore, the consumption of raw camel milk may contribute to the spread of human brucellosis in endemic regions.


Assuntos
Brucella abortus/isolamento & purificação , Camelus/microbiologia , Leite/microbiologia , Animais , Brucella abortus/genética , Brucelose/diagnóstico , Brucelose/epidemiologia , Brucelose/veterinária , Primers do DNA , Microbiologia de Alimentos , Irã (Geográfico)/epidemiologia , Reação em Cadeia da Polimerase/veterinária
8.
Prev Vet Med ; 169: 104697, 2019 Aug 01.
Artigo em Inglês | MEDLINE | ID: mdl-31311638

RESUMO

Malignant theileriosis of sheep and goats caused by Theileria lestoquardi is considered to be among the most important tick borne diseases in the Sudan. Information on the prevalence of the disease in different parts of the Sudan is limited. The purpose of this study was to estimate the prevalence of the disease in five states of the Sudan using molecular and serological assays. A total of 393 blood and serum samples from clinically asymptomatic sheep were analysed using nested reverse line blot (nRLB) and loop mediated isothermal amplification (LAMP), as well as an enzyme-linked immunosorbent assay (ELISA). The results indicated a sero-prevalence of 33.8% while RLB and LAMP assays revealed molecular prevalences of 29.5 and 22.6% respectively. The prevalence of Theileria lestoquardi varied significantly according to the geographical origin of the infected animals, whereas age and gender did not have a significant effect. RLB data indicated that T. lestoquardi usually occurred as a co-infection with the non-pathogenic Theileria ovis. Using RLB as a gold standard, a sensitivity of 68.1% and a specificity of 96.4% were recorded for LAMP and a sensitivity of 75.9% and a specificity of 83.8% for ELISA. The Kappa coefficient between nRLB and LAMP indicated a significant level of agreement (0.692), but only moderate concordance (0.572) between nRLB and ELISA. The results of the present study confirm and extend earlier findings regarding the widespread of T. lestoquardi infections in sheep in the Sudan. The data provide evidence that should enable the veterinary authorities to deploy appropriate control measures.


Assuntos
Doenças dos Ovinos/epidemiologia , Doenças dos Ovinos/parasitologia , Theileria/isolamento & purificação , Theileriose/epidemiologia , Animais , Ensaio de Imunoadsorção Enzimática/veterinária , Feminino , Geografia , Masculino , Reação em Cadeia da Polimerase/veterinária , Prevalência , Ovinos , Doenças dos Ovinos/sangue , Sudão/epidemiologia , Theileriose/sangue
9.
Prev Vet Med ; 169: 104712, 2019 Aug 01.
Artigo em Inglês | MEDLINE | ID: mdl-31311647

RESUMO

Surra is a zoonotic disease caused by Trypanosoma evansi, affecting the health and production of the livestock significantly. There are several methods to diagnose this disease, which have different principles, sensitivity, and specificity. Among them, the serological techniques using T. evansi as antigen are powerful tools for its epidemiological surveillance. However, they are poorly used due to inefficient in vitro propagation of T. evansi, which requires the use of laboratory animals for antigen production. In the present study, whole cell lysate of T. brucei brucei propagated in vitro was used as an antigen for the detection of anti-T. evansi immunoglobulin G in cattle through an indirect-ELISA. Based on a total of 45 samples from non-infected and 45 samples from T. evansi infected cattle, the sensitivity and specificity were estimated as 100% and 97.7%, respectively. After the validation, serological and molecular surveys were carried out in 710 cattle samples from two endemic Colombian regions (Antioquia and Arauca departments) for T. evansi where molecular prevalences of ˜7.0% were detected through the year and sporadic outbreaks of T. vivax infections have been associated to low prevalence of this species (<1%). A total of 424 (59.7%) samples were positive by indirect-ELISA T. b. brucei, while PCR test for T. evansi and T. vivax, showed 49 (6.9%) and no positive samples, respectively. Interestingly, categories of animals aged>1 year, Bos taurus breed, and those raised under intensive farming system exhibited a higher seroprevalence to T. evansi (P < 0.05). The results displayed a new alternative for antibody detection anti-T. evansi in livestock, using parasites propagated in vitro as antigen, which presents the advantage of higher standardization potential, and avoid the use of live animal for antigen production. A larger availability of this ELISA will generate useful information for a better understanding of the epidemiologic aspects, as well as for the management and control of these diseases in Colombia. However, the ability of the test to detect and/or cross react with T. vivax infections remains to be investigated.


Assuntos
Antígenos de Protozoários/isolamento & purificação , Ensaio de Imunoadsorção Enzimática/veterinária , Reação em Cadeia da Polimerase/veterinária , Trypanosoma brucei brucei/isolamento & purificação , Animais , Bovinos , Colômbia , Ensaio de Imunoadsorção Enzimática/métodos , Imunoglobulina G , Gado , Reação em Cadeia da Polimerase/métodos
10.
Onderstepoort J Vet Res ; 86(1): e1-e8, 2019 Jun 13.
Artigo em Inglês | MEDLINE | ID: mdl-31291731

RESUMO

Tick-borne diseases (TBDs) caused by Theileria, Babesia, Anaplasma and Ehrlichia species are common in tropical and subtropical regions. In this study, we investigated the presence and genetic diversity of Theileria spp., Anaplasma ovis, B. ovis, E. ruminantium and Anaplasma spp. in sheep from the Machakos and Homa Bay counties of Kenya. In order to improve the diagnosis and control of ovine TBDs, a total of 76 blood samples from apparently healthy sheep were screened using a polymerase chain reaction (PCR). The assays were conducted using primers based on Theileria spp. 18S rRNA, Anaplasma ovis Major surface protein-4 (AoMSP4), B. ovis 18S rRNA, E. ruminantium pCS20 and Anaplasma spp. 16S rRNA. The overall infection rates for Theileria spp., A. ovis, E. ruminantium and Anaplasma spp. were 39/76 (51.3%), 26/76 (34.2%), 6/76 (7.9%) and 31/76 (40.8%), respectively. The overall co-infection was 47/76 (61.8%). All Theileria spp. positive samples were confirmed to be of Theileria ovis on sequencing. A phylogenetic analysis of the 18S rRNA gene sequences of T. ovis revealed that all isolates of this study clustered with T. ovis sequences extracted from the GenBank suggesting this gene is highly conserved. E. ruminantium pCS20 sequences were in the same clade on the phylogenetic tree. However, three AoMSP4 sequences from this study appeared in the same clade, while one sequence formed a separate branch revealing genetic divergence. The 16S rRNA sequencing revealed uncharacterised Anaplasma spp. and A. ovis. The phylogenetic analyses of the uncharacterised Anaplasma spp. revealed that the two sequences from this study appear in an independent clade from other sequences extracted from the GenBank. This study provides important information regarding the occurrence of tick-borne pathogens and their degree of genetic diversity among sheep in Kenya, which is useful for the diagnosis and control of TBDs.


Assuntos
Doenças dos Ovinos/epidemiologia , Doenças Transmitidas por Carrapatos/veterinária , Anaplasma/genética , Anaplasma/isolamento & purificação , Anaplasmose/epidemiologia , Anaplasmose/virologia , Animais , Ehrlichia/genética , Ehrlichia/isolamento & purificação , Ehrlichiose/epidemiologia , Ehrlichiose/veterinária , Ehrlichiose/virologia , Feminino , Variação Genética , Quênia/epidemiologia , Masculino , Filogenia , Reação em Cadeia da Polimerase/veterinária , Ovinos , Doenças dos Ovinos/microbiologia , Theileria/genética , Theileria/isolamento & purificação , Theileriose/epidemiologia , Theileriose/microbiologia , Doenças Transmitidas por Carrapatos/epidemiologia , Doenças Transmitidas por Carrapatos/microbiologia
11.
J Vet Diagn Invest ; 31(4): 634-639, 2019 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-31179891

RESUMO

A cluster of 4 bovine abortions caused by Coxiella burnetii occurred in a dairy herd in Uruguay during a 2-mo period. Case 1 consisted of a placenta from an aborted cow; cases 2-4 were fetuses and their placentas. Grossly, the placenta from one aborted cow had moderate, diffuse reddening of the cotyledons and loss of translucency of the intercotyledonary areas. No gross lesions were observed in the other 3 placentas. Microscopically, 2 of 4 placentas had fibrinonecrotizing placentitis with abundant intratrophoblastic gram-negative coccobacilli. C. burnetii was identified intralesionally by immunohistochemistry (IHC) in all 4 placentas, and by PCR and DNA sequencing in 3 placentas analyzed by these techniques. One fetus had mild neutrophilic alveolitis with multinucleate syncytial cells; no gross or microscopic lesions were observed in the other 2 fetuses examined. The lungs of the 3 fetuses were negative for C. burnetii by IHC. Tests performed to investigate other possible causes of abortions in the 4 cases were negative. C. burnetii causes Q fever in humans and coxiellosis in animals. Clusters of abortions in cattle by C. burnetii have not been reported previously, to our knowledge; this bacterium has been considered an opportunistic pathogen associated only with sporadic abortion in cattle. We present herein a cluster of 4 bovine abortions caused by C. burnetii in a dairy farm during a period of 2 mo and a review of the literature on C. burnetii infection in cattle.


Assuntos
Aborto Animal/microbiologia , Doenças dos Bovinos/microbiologia , Coxiella burnetii/isolamento & purificação , Complicações Infecciosas na Gravidez/veterinária , Febre Q/veterinária , Aborto Animal/epidemiologia , Animais , Bovinos , Coxiella burnetii/genética , Feminino , Feto/microbiologia , Feto/patologia , Humanos , Imuno-Histoquímica , Placenta/microbiologia , Placenta/patologia , Reação em Cadeia da Polimerase/veterinária , Gravidez , Complicações Infecciosas na Gravidez/epidemiologia , Complicações Infecciosas na Gravidez/microbiologia , Febre Q/complicações , Febre Q/epidemiologia , Febre Q/microbiologia , Uruguai/epidemiologia
12.
Parasitol Res ; 118(8): 2419-2429, 2019 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-31230161

RESUMO

From September 2012 to May 2018, blood samples from 364 raptors (mostly adults) were collected and screened for trypanosomes and haemosporidians by microscopic examination and nested polymerase chain reactions (PCR). Trypanosoma spp. were identified in 15 birds from eight different species. Light microscopy revealed 14 cases of infection with Trypanosoma cf. corvi, including one each in black-shouldered kite (Elanus caeruleus, n = 49), Brahminy kite (Haliastur indus, n = 50), and spotted owlet (SO, Athene brama, n = 27); two mountain hawk-eagles (Spizaetus nipalensis, n = 3); and three each in Asian barred owlets (ABO, Glaucidium cuculoides, n = 27), barn owls (BO, Tyto alba, n = 65) and collared scops owls (CSO, Otus lettia, n = 41). In addition, one case of infection with T. avium was identified in an oriental scops owl (OSO, Otus sunia, n = 2). All infected raptors showed very low parasitemia levels. The PCR detected more three positives in one CSO, one Japanese sparrowhawk (Accipiter gularis), and one OSO. The sensitivity and specificity of the PCR method were 93.3% and 99.1%, respectively. The overall infection rate was very low (4.9%). The highest infection rate was recorded in cold-dry season (9.9%). Coinfection of Plasmodium with trypanosomes was found in all three ABOs. Coinfection with Haemoproteus spp. was found in one BO, three CSOs, and one SO. Coinfection with Haemoproteus spp. and Leucocytozoon danilewskyi was found in the OSO. Microfilarias were detected in one ABO and one CSO. The ultrastructure of trypomastigotes of T. cf. corvi in an ABO revealed fine structures. All small subunit ribosomal RNA (SSU rRNA) sequences belong to two clades: T. avium and T. corvi-culicavium complex/group. SSU rRNA gene amplification was not successful in one BO. The raptors with trypanosome infections showed normal hematological values and healthy appearance. Furthermore, this is the first report of T. avium in a nocturnal raptor from Thailand.


Assuntos
Doenças das Aves/parasitologia , Aves Predatórias/parasitologia , Trypanosoma/crescimento & desenvolvimento , Trypanosoma/genética , Tripanossomíase/veterinária , Animais , Haemosporida/genética , Haemosporida/isolamento & purificação , Plasmodium/genética , Plasmodium/isolamento & purificação , Reação em Cadeia da Polimerase/veterinária , Aves Predatórias/classificação , Tailândia , Trypanosoma/classificação , Trypanosoma/isolamento & purificação , Tripanossomíase/parasitologia
13.
J S Afr Vet Assoc ; 90(0): e1-e5, 2019 May 30.
Artigo em Inglês | MEDLINE | ID: mdl-31170779

RESUMO

Equid herpesvirus type 1 is primarily a respiratory tract virus associated with poor athletic performance that can also cause late gestation abortion, neonatal foal death and encephalomyelopathy. Horizontal transmission is well described, whereas evidence of vertical transmission of equid herpesvirus type 1 associated with the birth of a healthy foal has not been demonstrated. This study sampled a population of Thoroughbred mares (n = 71), and their healthy neonatal foals and foetal membranes, to test for the presence of both equid herpesvirus types 1 and 4 using a quantitative polymerase chain reaction assay. Foetal membrane swabs and tissue samples were taken immediately post-partum, and venous blood samples and nasal swabs were obtained from both mare and foal 8 h after birth. Neither equid herpesvirus type 1 nor equid herpesvirus type 4 nucleic acid was detected in any sample, and it was concluded that there was no active shedding of equid herpesvirus types 1 and 4 at the time of sampling. Consequently, no evidence of vertical transmission of these viruses could be found on this stud farm during the sampling period.


Assuntos
Animais Recém-Nascidos/virologia , Infecções por Herpesviridae/veterinária , Herpesvirus Equídeo 1/isolamento & purificação , Herpesvirus Equídeo 4/isolamento & purificação , Doenças dos Cavalos/virologia , Animais , Sangue/virologia , Feminino , Infecções por Herpesviridae/transmissão , Doenças dos Cavalos/transmissão , Cavalos , Transmissão Vertical de Doença Infecciosa/veterinária , Mucosa Nasal/virologia , Placenta/virologia , Reação em Cadeia da Polimerase/veterinária , Gravidez , África do Sul/epidemiologia
14.
J Med Microbiol ; 68(7): 1003-1011, 2019 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-31172912

RESUMO

PURPOSE: The present study aimed to establish pretreatment protocols as well as real-time and droplet digital polymerase chain reaction (PCR) methodologies to detect and quantify Erysipelothrix rhusiopathiae (ER) DNA in blood samples from infected chickens, as tools for routine diagnostics and monitoring of experimental infections. Chicken blood is a problematic matrix for PCR analysis because nucleated erythrocytes contribute large amounts of host DNA that inhibit amplification. METHODOLOGY: Using artificially spiked samples of fresh chicken blood, as well as blood samples from three experimental infection studies, the performance of pretreatment protocols, including choice of blood stabilization agent, centrifugation speeds and Ficoll gradient separation, was evaluated. The results were compared with those from traditional culture-based protocols combined with matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS).Results/Key findings. Simple preparations producing cell-free samples performed well on artificial spike-in samples, providing high sensitivity. However, performance was poor in clinical samples or artificial samples where the bacteria were incubated for 4 h or more in fresh blood prior to DNA extraction. In these samples, a Ficoll separation protocol that creates samples rich in lymphocytes, monocytes and thrombocytes prior to DNA extraction was far more effective. CONCLUSIONS: Our results indicate that ER bacteria undergo rapid phagocytosis in chicken blood and that analysis of a blood fraction enriched for phagocytic cells is necessary for reliable detection and quantification. The presented results explain the poor performance of PCR detection reported in previously published experimental ER infection studies, and the proposed solutions are likely to have broader implications for PCR-based veterinary diagnostics in non-mammalian host species such as poultry and fish.


Assuntos
Galinhas/microbiologia , DNA Bacteriano/genética , Infecções por Erysipelothrix/microbiologia , Erysipelothrix/genética , Reação em Cadeia da Polimerase/veterinária , Doenças das Aves Domésticas/microbiologia , Animais , Erysipelothrix/isolamento & purificação , Infecções por Erysipelothrix/diagnóstico , Eritrócitos/citologia , Eritrócitos/microbiologia , Reação em Cadeia da Polimerase/métodos
15.
Microbiol Immunol ; 63(8): 328-333, 2019 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-31209913

RESUMO

Ticks are one of the arthropods that play an important role in the transmission of numerous pathogens to livestock and humans. We investigated the presence of tick-borne bacteria in 23 Amblyomma varanense that fed on a water monitor (Varanus salvator) in Indonesia. Anaplasmataceae and borreliae were detected by PCR in 17.4% and 95.7% of ticks, respectively. "Candidatus Rickettsia sepangensis", spotted fever group of Rickettsia, was detected in 21.7% of ticks. The water monitor is a common reptile that is widely encountered in city areas in Asian countries. Our results suggested that Am. varanense on water monitor in Indonesia harbored several kinds of bacteria.


Assuntos
Anaplasma/isolamento & purificação , Borrelia/isolamento & purificação , Ixodidae/microbiologia , Lagartos/microbiologia , Rickettsia/isolamento & purificação , Anaplasma/classificação , Anaplasma/genética , Animais , Borrelia/classificação , Borrelia/genética , DNA Bacteriano , DNA Ribossômico/genética , Feminino , Indonésia , Masculino , Filogenia , Reação em Cadeia da Polimerase/métodos , Reação em Cadeia da Polimerase/veterinária , Rickettsia/classificação , Rickettsia/genética , Análise de Sequência de DNA/veterinária , Doenças Transmitidas por Carrapatos/microbiologia
17.
Parasitol Res ; 118(7): 2079-2086, 2019 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-31187226

RESUMO

Cryptosporidiosis of calves is caused by the enteroprotozoan Cryptosporidium spp. The disease results in intense diarrhea of calves associated with substantial economic losses in dairy farming worldwide. The aim of this study was to determine calf, herd, and within-herd Cryptosporidium prevalence and identify Cryptosporidium species and subtypes in calves with diarrhea in intensive dairy herds in central Argentina. A total of 1073 fecal samples were collected from 54 randomly selected dairy herds. Cryptosporidium-oocysts were isolated and concentrated from fecal samples using formol-ether and detected by light microscopy with the modified Ziehl-Neelsen technique. Overall prevalence of oocyst-excreting calves was found to be 25.5% (274/1073) (95% C.I. 22.9; 28.1%). Of the herds studied, 89% (48/54) included at least one infected calf, whereas within-herd prevalence ranged from the absence of infection to 57% (20/35). A highly significant association was found between the presence of diarrhea and C. parvum infection (χ2 = 55.89, p < 0.001). For species determination, genomic DNA isolated from oocyst-positive fecal samples was subjected to PCR-RFLP of the 18S rRNA gene resulting exclusively in Cryptosporidium parvum identification. C. parvum isolates of calves displaying diarrhea and high rate of excretion of oocysts were subtyped by PCR amplification and direct sequencing of the 60 kDa glycoprotein (GP60) gene. Altogether five GP60 subtypes, designated IIaA18G1R1, IIaA20G1R1, IIaA21G1R1, IIaA22G1R1, and IIaA24G1R1 were identified. Interestingly, IIaA18G1R1 and IIaA20G1R1 were predominant in calves with diarrhea and high infection intensity. Notably, IIaA24G1R1 represents a novel, previously unrecognized C. parvum subtype. The subtype IIaA18G1R1, frequently found in this study, is strongly implicated in zoonotic transmission. These results suggest that calves might be an important source for human cryptosporidiosis in Argentina.


Assuntos
Doenças dos Bovinos/epidemiologia , Criptosporidiose/epidemiologia , Cryptosporidium parvum/classificação , Cryptosporidium/classificação , Diarreia/veterinária , Animais , Argentina/epidemiologia , Bovinos , Doenças dos Bovinos/parasitologia , Criptosporidiose/parasitologia , Criptosporidiose/transmissão , Cryptosporidium/genética , Cryptosporidium/isolamento & purificação , Cryptosporidium parvum/genética , Cryptosporidium parvum/isolamento & purificação , Diarreia/epidemiologia , Diarreia/parasitologia , Fezes/parasitologia , Feminino , Glicoproteínas/genética , Humanos , Oocistos , Reação em Cadeia da Polimerase/veterinária , Polimorfismo de Fragmento de Restrição , Prevalência , Zoonoses
18.
Onderstepoort J Vet Res ; 86(1): e1-e6, 2019 May 28.
Artigo em Inglês | MEDLINE | ID: mdl-31170781

RESUMO

Reports on the occurrence of Campylobacter spp. in dogs in South Africa are non-existent. This study investigated the prevalence of Campylobacter spp. in 481 dogs visiting four rural community veterinary clinics in South Africa. Dogs were screened for Campylobacter spp. by culture and polymerase chain reaction (PCR), and logistic regression analysis was performed to assess the association between sex, clinic, breed and age and the occurrence of Campylobacter spp. in dogs. The prevalence of Campylobacter spp. was 41.50% (95% confidence interval [CI], 37.39% - 46.04%). Campylobacter jejuni, C. upsaliensis and C. coli were detected in 29.31% (95% CI, 25.42% - 33.54%), 13.10% (95% CI, 10.37% - 16.42%) and 5.41% (95% CI, 3.71% - 7.82%) of dogs, respectively. Dogs carrying more than one species of Campylobacter spp. accounted for 6.23% (95% CI, 4.40% - 8.78%). Campylobacter upsaliensis and C. jejuni were detected in 3.74% (95% CI, 2.37% - 5.86%), whereas C. coli and C. jejuni were found in 2.49% (95% CI, 1.42% - 4.34%) of dogs. Age and clinic were the risk factors significantly associated with Campylobacter spp. occurrence, while age, breed and clinic were predictors of C. jejuni carriage. Furthermore, age was the only risk factor associated with a higher likelihood of carrying C. upsaliensis. The prevalence of Campylobacter spp. C. jejuni and C. upsaliensis increased significantly as dogs grew older. In addition, the odds of carrying Campylobacter spp. were higher in the Staffordshire bull terrier breed compared to crossbreed dogs. In conclusion, this study shows that dogs visiting rural community veterinary clinics in South Africa are reservoirs of Campylobacter spp. and may be potential sources of Campylobacter spp. for humans living in close proximity of the dog populations under study.


Assuntos
Infecções por Campylobacter/veterinária , Campylobacter jejuni/isolamento & purificação , Campylobacter upsaliensis/isolamento & purificação , Doenças do Cão/epidemiologia , Animais , Infecções por Campylobacter/epidemiologia , Infecções por Campylobacter/etiologia , Infecções por Campylobacter/prevenção & controle , Campylobacter jejuni/genética , Campylobacter upsaliensis/genética , Estudos Transversais , Doenças do Cão/etiologia , Doenças do Cão/prevenção & controle , Cães , Feminino , Hospitais Veterinários , Masculino , Reação em Cadeia da Polimerase/veterinária , Prevalência , Fatores de Risco , População Rural , África do Sul/epidemiologia
19.
Rev Bras Parasitol Vet ; 28(2): 320-324, 2019 Jun 06.
Artigo em Inglês | MEDLINE | ID: mdl-31188944

RESUMO

Trypanosomiasis caused by Trypanosoma evansi can seriously affect both domestic and wild animals. This article reports on an outbreak of canine trypanosomiasis on a farm in the Pantanal region of Brazil. The farm had 38 dogs, 20 of which died before receiving veterinary care. The remaining 18 dogs were underwent anamnesisn, clinical examination, hematological and biochemical evaluations. Blood smears and PCR analysis were performed for the diagnosis. The treatment protocols used according to the clinical recovery or parasitological cure of the dogs, using diminazene diaceturate, isometamidium chloride or quinapyramine sulfate. Post-treatment parasitological evaluation was performed by the microhematocrit technique. 7/18 dogs were PCR positive for T. evansi (confirmed by sequencing). There was clinical findings, which were consistent with both the acute and chronic stages of the disease in dogs. The infected dogs all exhibited at least one clinical sign of the disease. The hematological findings were compatible with trypanosomiasis, highlighting the hypochromic microcytic anemia as the main outcome. No treatment protocol was fully effective and the prolonged use of diminazene diaceturate caused the death of an animal. The trypanosomiasis can cause high rates of morbidity and mortality in dogs and difficulty in establishment an effective and safe therapeutic protocol.


Assuntos
Diminazena/análogos & derivados , Doenças do Cão/diagnóstico , Fenantridinas/uso terapêutico , Compostos de Quinolínio/uso terapêutico , Tripanossomíase/diagnóstico , Animais , Brasil/epidemiologia , Diminazena/uso terapêutico , Surtos de Doenças , Doenças do Cão/tratamento farmacológico , Doenças do Cão/epidemiologia , Cães , Feminino , Masculino , Reação em Cadeia da Polimerase/veterinária , Tripanossomíase/tratamento farmacológico , Tripanossomíase/epidemiologia
20.
Rev Bras Parasitol Vet ; 28(2): 303-305, 2019 Jun 13.
Artigo em Inglês | MEDLINE | ID: mdl-31215604

RESUMO

Leishmania spp. are important agents of human and animal leishmaniases that have an important impact on public health. In this study, we aimed to detect the circulation of Leishmania spp. in cattle from a visceral leishmaniasis non-endemic area of the state of São Paulo, Brazil. DNA was extracted from blood samples from 100 heifers in the municipality of Pirassununga and was amplified using primers specific for the first internal transcriber spacer (ITS1), to assess the presence of trypanosomatids. The assays revealed that one sample presented bands of between 300 and 350 base pairs. In GenBank, this sample matched 100% with Leishmania infantum (314 base pairs). The results suggest that cattle can be infected by Leishmania infantum in Brazil.


Assuntos
Doenças dos Bovinos/diagnóstico , DNA Espaçador Ribossômico/genética , Leishmania infantum/genética , Leishmaniose Visceral/veterinária , Animais , Bovinos , Leishmaniose Visceral/diagnóstico , Reação em Cadeia da Polimerase/veterinária
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