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1.
Comp Immunol Microbiol Infect Dis ; 78: 101689, 2021 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-34225227

RESUMO

Leptospirosis is the most widespread zoonosis worldwide, and it can cause reproductive failures in livestock, while in humans may vary from a mild fever to multi-organ failure and death. Due to this, in this study, we evaluated the usefulness of the segment encoding LigB C-terminus region, only present in pathogenic as target for a diagnostic PCR. This new PCR yielded a 100 % positivity for pathogenic Leptospira species and no cross-reactivity was found with intermediate or non-pathogenic species, or with other microorganisms, demostrating its high analytical specificity. The estimated analytical sensitivity was higher in serum samples than in blood or urine samples (6-9 × 102 lept/mL and 6-9 × 105 and 6-9 × 106 lept/mL, respectively). Multiple sequence alignment of the target region from different pathogenic Leptospira species confirmed that this gene region is highly conserved among these species, with few single nucleotide polymorphisms. The ligb-ct PCR here developed appears as a useful tool for the molecular diagnosis of leptospirosis.


Assuntos
Leptospira , Leptospirose , Animais , Leptospira/genética , Leptospirose/diagnóstico , Leptospirose/veterinária , Fases de Leitura Aberta , Reação em Cadeia da Polimerase/veterinária , Zoonoses
2.
Vet Ital ; 57(1): 29-39, 2021 May 11.
Artigo em Inglês | MEDLINE | ID: mdl-34313096

RESUMO

Marek's disease (MD) is one of the most significant neoplastic diseases of poultry caused by Marek's disease virus (MDV), an oncogenic avian herpesvirus which is responsible for great economic losses to the poultry industry worldwide. MD is being manifested as an acute disease with lymphomas in multiple visceral organs. In the present study, an outbreak of MD was investigated in one of the poultry farms from Andhra Pradesh, India. The gross lesions in the affected birds included lymphomas in different visceral organs like liver, spleen, proventriculus, heart and ovaries. Histopathology revealed presence of uniform lymphoblastoid cell infiltration typical of MD. The isolation of the virus was carried out in duck embryo fibroblast cells. After three blind passages, the cell cultures revealed plaque formation typical of MDV. Further confirmation of the virus was carried out by PCR targeting 132 bp repeats of serotype­1 MDV and the oncogenes Meq and vIL­8 were amplified and sequenced. The nucleotide and phylogenetic analysis of the virus confirmed the virus as virulent serotype­ 1 MDV. The present outbreak suggests the need for change in the vaccination regimen of MD vaccination with appropriate serotype­ 1 MD vaccines in Indian poultry flocks as the HVT and bivalent vaccines are unable to protect the flocks against virulent MDV.


Assuntos
Surtos de Doenças/veterinária , Herpesvirus Galináceo 2/isolamento & purificação , Doença de Marek/epidemiologia , Doenças das Aves Domésticas/epidemiologia , Aves Domésticas , Animais , Surtos de Doenças/prevenção & controle , Índia/epidemiologia , Doença de Marek/virologia , Reação em Cadeia da Polimerase/veterinária , Doenças das Aves Domésticas/virologia , Vacinação/veterinária
3.
Vet Ital ; 57(1): 83-87, 2021 May 11.
Artigo em Inglês | MEDLINE | ID: mdl-34313103

RESUMO

Common pathogens of intensive poultry farms, either parasitic or bacterial, such as Coccidiaor Salmonella, are well known and strictly controlled by veterinary management. This case study reports an unusual case of runting stunting syndrome (RSS) observed on a Sicilian poultry farm of broiler chickens during 2019. The investigation was carried out on five chickens which present delayed in body weight and growth performance. Animals showed also difficulty in deambulation and diarrhea. At necropsy, intestinal lesions were detected in three of the five clinical cases. Gut samples were collected and analyzed to identify potential pathogens responsible for the RSS. Presence of viruses was detected by using quantitative reverse transcription PCR (RT­qPCR), while selected tissues were fixed and embedded in paraffin wax according to routine procedures. All histological sections were stained with hematoxylin­eosin. RT­qPCR successfully detected both Chicken astrovirus (CAstV) and Avian orthoreovirus (ARV). Histology evidenced severe specific lesions on the intestinal mucosa in liver and kidneys. Chicken astrovirus and Avian orthoreovirus RNA was also detected in cecal tonsils, kidney and liver, thus implying their possible primary role in inducing the disease. Further studies are needed to evaluate the role of other possible factors (low biosecurity measures, e.g.) and, most of all, the consequences in terms of economic losses and animal health impairment.


Assuntos
Infecções por Astroviridae/veterinária , Avastrovirus/isolamento & purificação , Galinhas , Orthoreovirus Aviário/isolamento & purificação , Doenças das Aves Domésticas/diagnóstico , Infecções por Reoviridae/veterinária , Animais , Infecções por Astroviridae/complicações , Infecções por Astroviridae/diagnóstico , Avastrovirus/genética , Coinfecção , Diagnóstico Diferencial , Orthoreovirus Aviário/genética , Reação em Cadeia da Polimerase/veterinária , Doenças das Aves Domésticas/virologia , RNA Viral/análise , Infecções por Reoviridae/complicações , Infecções por Reoviridae/diagnóstico , Sicília
4.
Rev Bras Parasitol Vet ; 30(3): e009121, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-34259743

RESUMO

The dog is the main domestic reservoir of Leishmania and font of infection for the vector, constituting an important host for the transmission of the parasite to humans. Non-invasive collection of swab samples for leishmaniasis diagnosis has been a promising alternative. This study analyzed the positivity of polymerase chain reaction (PCR) for the diagnosis of canine leishmaniasis in conjunctiva samples. DNA extraction was performed using SDS 20% and PCR was performed using 13A/13B primers that amplify 120-bp of Leishmania kDNA. Of the 77 dogs analyzed, 50 (64.93%) had ocular changes: 25 (32.47%) dogs had periocular lesion, 41 (53.25%) dogs had purulent eye discharge, and 17 (22.08%) dogs had both signals. PCR was positive in 35 dogs (45.45%), and there was no significant difference between dogs with and without ocular signals (p=0.4074). PCR positivity was significant higher in dogs without periocular injury (p=0.0018). Conjunctive PCR, a less invasive, fast, and painless collection technique, is indicated to complement the diagnosis, especially in dogs without periocular injury, independent of the presence of purulent eye discharge.


Assuntos
Doenças do Cão , Leishmania infantum , Leishmania , Leishmaniose Visceral , Animais , Túnica Conjuntiva , DNA , DNA de Protozoário/genética , Doenças do Cão/diagnóstico , Cães , Leishmania/genética , Leishmania infantum/genética , Leishmaniose Visceral/veterinária , Reação em Cadeia da Polimerase/veterinária
5.
Theriogenology ; 172: 300-306, 2021 Sep 15.
Artigo em Inglês | MEDLINE | ID: mdl-34311221

RESUMO

Campylobacter fetus is a zoonotic pathogen found in cattle, in which it is one of the main causes of infectious infertility. Most diagnostic laboratories use PCR as quick easy tool for C. fetus identification. However, there is no standardized PCR assay for C. fetus detection and subspecies differentiation, hindering the comparison of results. In this study, we evaluated selected PCR assays targeting the 16S rRNA, gyrB, cpn60, cstA, cdtB and nahE genes for C. fetus identification and ISCfe1, sapB2, parA and virB11 for subspecies differentiation. Analytical sensitivity and specificity were assessed for each PCR assay, and the assays were then tested on 289 bull preputial samples that had also been analysed by 16S rRNA barcode metagenomics. In total, 41 C. fetus-positive samples were included. The P12 PCR assay targeting the gyrB gene performed best, detecting the pathogen in 95.1% of positive samples. For the discrimination of C. fetus subspecies, we were able to identify a proportion (85.4%) of the C. fetus-positive samples correctly as C. fetus venerealis with at least one subspecies-specific PCR, but C. fetus fetus was not detected in any of the samples tested. Remarkably, C. fetus subspecies amplification was observed following PCR on some samples (33.1%) considered C. fetus-negative, highlighting the need for rigorous criteria for discriminating between C. fetus subspecies, to improve understanding of the role of the two C. fetus subspecies in the epidemiology and pathogenesis of bovine infectious infertility.


Assuntos
Infecções por Campylobacter , Doenças dos Bovinos , Animais , Infecções por Campylobacter/diagnóstico , Infecções por Campylobacter/veterinária , Campylobacter fetus/genética , Bovinos , Doenças dos Bovinos/diagnóstico , Feto , Masculino , Reação em Cadeia da Polimerase/veterinária , RNA Ribossômico 16S/genética
6.
Ticks Tick Borne Dis ; 12(5): 101708, 2021 09.
Artigo em Inglês | MEDLINE | ID: mdl-34175736

RESUMO

Cattle fever ticks, Rhipicephalus microplus and R. annulatus have been eradicated from the United States and inspectors from the U.S. Department of Agriculture (USDA), Animal Plant Health Inspection Service (APHIS), Cattle Fever Tick Eradication Program (CFTEP) monitor the quarantine zone along the Texas border to prevent the introduction of livestock carrying cattle fever ticks from Mexico. Stray livestock apprehended by CFTEP in the zone are checked for ticks and tested for infectious disease-causing pathogens but are not evaluated for evidence of infection with tick-borne pathogens. We tested blood samples collected from stray cattle by CFTEP inspectors for evidence of infection with tick-borne pathogens. As a comparison group representing U.S. resident cattle, we tested blood samples that had been sent to the Texas A&M Veterinary Medical Diagnostic Laboratory (TVMDL) for unrelated testing. Both sets of blood samples were evaluated using the same specific and broad-spectrum PCR assays. For the border cattle the overall prevalence of infection with one or more tick-borne pathogen was 58.5 % (79/135) with many co-infections, including 30 cattle positive for Babesia bovis and/or Babesia bigemina (22.2 %) and 77 cattle positive for Anaplasma marginale (57 %), three of these animals were also positive for Borrelia theileri. No resident cattle represented by the TVMDL samples were infected with either of the Babesia spp., or with Borrelia theileri, but three were positive for Theileria orientalis and 7.3 % (7/96) were positive for A. marginale. These data show that cattle originating in Mexico have a higher prevalence of infection with tick-borne pathogens relative to resident U.S. cattle and specifically, a proportion are infected with bovine Babesia, which is absent from U.S. cattle populations. Consequently, these stray cattle may be a reservoir of tick-borne pathogens and if populations of Boophilus ticks become reestablished in areas where they had previously been eradicated, could pose a significant risk to the U.S. Cattle industry.


Assuntos
Anaplasmose/epidemiologia , Babesiose , Doenças dos Bovinos/epidemiologia , Coccidiose/veterinária , Doenças Transmitidas por Carrapatos/epidemiologia , Anaplasma/isolamento & purificação , Anaplasma marginale/isolamento & purificação , Animais , Vetores Aracnídeos/microbiologia , Vetores Aracnídeos/parasitologia , Babesia/isolamento & purificação , Babesiose/epidemiologia , Borrelia/isolamento & purificação , Bovinos , Doenças dos Bovinos/microbiologia , Doenças dos Bovinos/parasitologia , Coccidiose/epidemiologia , Reservatórios de Doenças/microbiologia , Reservatórios de Doenças/parasitologia , Vetores de Doenças , México , Reação em Cadeia da Polimerase/veterinária , Infecções Protozoárias em Animais/epidemiologia , Rhipicephalus/microbiologia , Rhipicephalus/parasitologia , Texas , Theileria/isolamento & purificação , Theileriose/epidemiologia
7.
Parasitol Res ; 120(7): 2343-2350, 2021 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-34110501

RESUMO

Arthropod vectors are frequently exposed to a diverse assemblage of parasites, but the consequence of these infections on their biology and behavior are poorly understood. We experimentally evaluated whether the ingestion of a common protozoan parasite of avian hosts (Haemoproteus spp.; Haemosporida: Haemoproteidae) impacted the survivorship of Culex quinquefasciatus (Say) (Diptera: Culicidae). Blood was collected from wild northern cardinals (Cardinalis cardinalis) in College Station, Texas, and screened for the presence of Haemoproteus spp. parasites using microscopic and molecular methods. Experimental groups of Cx. quinquefasciatus mosquitoes were offered Haemoproteus-positive cardinal blood through an artificial feeding apparatus, while control groups received Haemoproteus-negative cardinal blood or domestic canary (Serinus canaria domestica) blood. Culex quinquefasciatus mosquitoes exposed to Haemoproteus infected cardinal blood survived significantly fewer days than mosquitoes that ingested Haemoproteus-negative cardinal blood. The survival of mosquitoes fed on positive cardinal blood had a median survival time of 18 days post-exposure and the survival of mosquitoes fed on negative cardinal blood exceeded 50% across the 30 day observation period. Additionally, mosquitoes that fed on canary controls survived significantly fewer days than cardinal negative controls, with canary control mosquitoes having a median survival time of 17 days. This study further supports prior observations that Haemoproteus parasites can be pathogenic to bird-biting mosquitoes, and suggests that Haemoproteus parasites may indirectly suppress the transmission of co-circulating vector-borne pathogens by modulating vector survivorship. Our results also suggest that even in the absence of parasite infection, bloodmeals from different bird species can influence mosquito survivorship.


Assuntos
Culex/fisiologia , Culex/parasitologia , Haemosporida/fisiologia , Mosquitos Vetores/fisiologia , Mosquitos Vetores/parasitologia , Passeriformes/parasitologia , Animais , Doenças das Aves/parasitologia , Doenças das Aves/transmissão , Canários/sangue , Canários/parasitologia , DNA de Protozoário/sangue , Passeriformes/sangue , Reação em Cadeia da Polimerase/veterinária , Probabilidade , Modelos de Riscos Proporcionais , Infecções Protozoárias em Animais/parasitologia , Infecções Protozoárias em Animais/transmissão , Texas
8.
J Equine Vet Sci ; 102: 103458, 2021 07.
Artigo em Inglês | MEDLINE | ID: mdl-34119210

RESUMO

Having considered that the current methods are costly and time-consuming, we designed an only 3 pairs primer-based PCR test to accurately identify the species and gender in horses, donkeys, mules and hinnies. Through a thorough sequence comparison between horse and donkey's highly similar genomes, and a vast amount of preliminary confirmation, we found that three fragments, CNGB3 gene on an autosome, displacement loop region on mitochondrial DNA and SRY genes on chromosome Y, within these equine DNA, are enough to enable us achieving our goal. The PCR test described here would be an economical, fast and accurate alternative for the most commonly-used methods, polymerase chain reaction-restriction fragment length polymorphism, microsatellite assay, and sequencing.


Assuntos
Equidae , Cromossomo Y , Animais , Equidae/genética , Cavalos/genética , Repetições de Microssatélites/genética , Reação em Cadeia da Polimerase/veterinária , Polimorfismo de Fragmento de Restrição
9.
Parasitol Res ; 120(7): 2631-2640, 2021 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-34152467

RESUMO

Bio-invasions are a major threat to biodiversity and ecosystems globally and may contribute to the proliferation of emerging infectious diseases. We examined the prevalence and phylogenetic diversity of avian haemosporidian parasites infecting the non-native house sparrows (Passer domesticus) and the native southern grey-headed sparrows (Passer diffusus). Blood samples from 104 sparrows (74 house sparrows and 30 southern grey-headed sparrows) mist-netted inside and around the Kruger National Park were used. Genomic DNA was extracted from each blood sample and subjected to nested PCR analyses, Sanger sequencing and phylogenetic analyses. Overall, 35.57% (37/104) of the birds sampled were infected with at least one haemosporidian parasites. Southern grey-headed sparrows had a higher parasite prevalence (60%) than house sparrows (24.3%). A total of 16 parasite lineages were identified, of which eight were novel lineages. Whereas Haemoproteus spp. showed the highest lineage diversity, Leucocytozoon spp. were the most prevalent parasites, albeit with significant differences between sparrow species. A single Plasmodium sp. infection was recorded in a southern grey-headed sparrow. In support of the enemy release hypothesis, we found that prevalence on non-native house sparrows was lower than prevalence recorded in their region of origin and also that they were infected only by indigenous parasites lineages.


Assuntos
Doenças das Aves/epidemiologia , Doenças das Aves/parasitologia , Haemosporida , Infecções por Protozoários/epidemiologia , Pardais/parasitologia , Animais , Ecossistema , Feminino , Haemosporida/genética , Espécies Introduzidas , Funções Verossimilhança , Masculino , Parasitemia/epidemiologia , Parasitemia/parasitologia , Parasitemia/veterinária , Filogenia , Plasmodium/genética , Reação em Cadeia da Polimerase/veterinária , Prevalência , Infecções por Protozoários/parasitologia , África do Sul/epidemiologia , Clima Tropical
10.
Comp Immunol Microbiol Infect Dis ; 77: 101677, 2021 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-34116387

RESUMO

OBJECTIVE: Toxoplasma gondii is a protozoan parasite that is widely prevalent in most warm-blooded vertebrates. Humans mainly become infected by eating raw or undercooked meat. This study was designed to investigate the infection of cattle with T. gondii in Jahrom, southern Iran. METHODS: Tissue samples consisting of heart, diaphragm, and tongue were collected from 125 slaughtered cattle. DNA samples were extracted from the homogenized tissues. T. gondii was detected and genotyped using nested-polymerase chain reaction (Nested-PCR) and polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) based on GRA6 and SAG2 (3', 5' terminal regions) genes, respectively. RESULTS: The prevalence of T. gondii DNA was 56% in cattle. The most infected tissue was the diaphragm (54.4%) followed by the heart (48.8%) and tongue (43.2%). Type II was the most prevalent genotype (70%) among T. gondii isolates. CONCLUSION: In this study, the high prevalence of T. gondii infection in cattle meat indicates the important role of cattle in the transmission of infection to humans. Therefore, incorporating the correct method of consuming meat in health education programs is crucial to prevent human infection.


Assuntos
Doenças dos Bovinos , Toxoplasma , Toxoplasmose Animal , Animais , Bovinos , Doenças dos Bovinos/epidemiologia , DNA de Protozoário/genética , Genótipo , Irã (Geográfico)/epidemiologia , Reação em Cadeia da Polimerase/veterinária , Polimorfismo de Fragmento de Restrição , Toxoplasma/genética , Toxoplasmose Animal/diagnóstico , Toxoplasmose Animal/epidemiologia
11.
BMC Vet Res ; 17(1): 191, 2021 May 13.
Artigo em Inglês | MEDLINE | ID: mdl-33985499

RESUMO

BACKGROUND: Tetratrichomonas gallinarum is parasitic protozoa with a wide host range. However, its lethal infection is rare reported. CASE PRESENTATION: Here, we described the first lethal cases of T. gallinarum infection in black swans in China. Five black swans died within a week in succession without obvious symptoms except mild diarrhea. At necropsy, severe lesions were observed in caeca with thickened caecal walls and hemorrhages in the mucosa. A large number of moving trophozoites were found in the contents of the cecum by microscopic examination. The livers were enlarged with multiple bleeding spots on the surface. Histopathology of the livers showed mononuclear cell infiltration and moderate hyperplasia of fibrous tissue. The histopathology of the cecum showed that the villi of the cecum were edematous. Finally, the presence of T. gallinarum was determined by specific PCR andin-situ hybridization assay. Additionally, common pathogens that can cause similar symptoms were excluded. CONCLUSIONS: The death of the black swan was caused by T. gallinarum, suggesting that the parasite might be a new threat to the Cygnus birds.


Assuntos
Doenças das Aves/parasitologia , Infecções Protozoárias em Animais/patologia , Trichomonadida/isolamento & purificação , Animais , Anseriformes , Doenças das Aves/patologia , Doenças do Ceco/parasitologia , Doenças do Ceco/patologia , China , Hibridização In Situ/veterinária , Fígado/parasitologia , Fígado/patologia , Reação em Cadeia da Polimerase/veterinária , Trichomonadida/genética
12.
Vet Res ; 52(1): 68, 2021 May 12.
Artigo em Inglês | MEDLINE | ID: mdl-33980312

RESUMO

Glaesserella parasuis strains were characterized by serotyping PCR, vtaA virulence marker Leader Sequence (LS)-PCR, clinical significance, and geographic region. Overall, the serovars 4, 5/12, 7, 1, and 13 were the most commonly detected. Serovars of greatest clinical relevance were systemic isolates that had a higher probability of being serovar 5/12, 13, or 7. In comparison, pulmonary isolates had a higher likelihood of being serovars 2, 4, 7, or 14. Serovars 5/12 and 13 have previously been considered disease-associated, but this study agrees with other recent studies showing that serovar 7 is indeed associated with systemic G. parasuis disease. Serovar 4 strains illustrated how isolates can have varying degrees of virulence and be obtained from pulmonary, systemic, or nasal sites. Serovars 8, 9, 15, and 10 were predominantly obtained from nasal samples, which indicates a limited clinical significance of these serovars. Additionally, most internal G. parasuis isolates were classified as virulent by LS-PCR and were disease-associated isolates, including serovars 1, 2, 4, 5/12, 7, 13, and 14. Isolates from the nasal cavity, including serovars 6, 9, 10, 11, and 15, were classified as non-virulent by LS-PCR. In conclusion, the distribution of G. parasuis serovars remains constant, with few serovars representing most of the strains isolated from affected pigs. Moreover, it was confirmed that the LS-PCR can be used for G. parasuis virulence prediction of field strains worldwide.


Assuntos
Infecções por Haemophilus/veterinária , Haemophilus parasuis/genética , Doenças dos Suínos/epidemiologia , Animais , Ásia/epidemiologia , Europa (Continente)/epidemiologia , Infecções por Haemophilus/epidemiologia , Infecções por Haemophilus/microbiologia , América do Norte/epidemiologia , Reação em Cadeia da Polimerase/veterinária , Prevalência , Estudos Soroepidemiológicos , Sorotipagem/veterinária , Sus scrofa , Suínos , Doenças dos Suínos/microbiologia
13.
J Vet Diagn Invest ; 33(4): 684-694, 2021 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-33955287

RESUMO

We developed a sandwich ELISA that detects Clostridium botulinum C and D toxins and reverse-transcription real-time PCRs (RT-rtPCRs) that detect botulinum C and D toxin genes, respectively, to replace the mouse bioassay. The toxin genes were closely associated with the toxin molecules and used as surrogates for the presence of toxin. Samples (638) from 103 clinical cases of birds (302) with suspected botulinum toxicity came from wild birds and poultry (9 cases). Samples tested included blood serum, other body fluids, various tissues, gut contents, maggots, water, and sediment. Botulism was diagnosed in 34 cases (all of which had positive samples in the ELISA, the C toxin gene RT-rtPCR, or both assays). Botulism was suspected in 16 cases (each of which had 1 positive sample either in the ELISA or the C toxin gene RT-rtPCR). In the remaining 53 cases, no samples were positive, but botulism could not be excluded in 32 of these cases, whereas there was no indication of botulism or another diagnosis in 21 cases. The D toxin gene was not detected in any of the clinical samples. No C or D toxin genes were detected in 71 pooled cloacal swabs from 213 healthy migratory birds. The use of an ELISA that detects botulinum C and D toxins in combination with a RT-rtPCR for the botulinum C toxin gene can help confirm the diagnosis of botulism in birds.


Assuntos
Doenças das Aves/diagnóstico , Toxinas Botulínicas/isolamento & purificação , Botulismo/veterinária , Patos , Ensaio de Imunoadsorção Enzimática/veterinária , Reação em Cadeia da Polimerase/veterinária , Animais , Animais Selvagens , Toxinas Botulínicas/genética , Botulismo/diagnóstico , Galinhas , Ensaio de Imunoadsorção Enzimática/métodos , Doenças das Aves Domésticas/diagnóstico , Austrália Ocidental
14.
Dis Aquat Organ ; 144: 175-185, 2021 May 06.
Artigo em Inglês | MEDLINE | ID: mdl-33955855

RESUMO

Systemic phaeohyphomycosis, aka 'fluid belly', is one of the most important emergent diseases in sturgeon Acipenser spp. aquaculture. The etiologic agent is the saprobic, dematiaceous fungus Veronaea botryosa. Effective vaccines and chemotherapeutic treatments are currently unavailable. Additionally, the fungus is a slow-growing organism, taking from 10-15 d for colonies to be observed in agar media. To this end, a specific quantitative PCR (qPCR) targeting the V. botryosa ß-tubulin gene was developed and validated. The specificity of the assay to V. botryosa was initially confirmed in silico and in vivo against common fungal fish pathogens, including closely related members of the order Chaetothyriales (Exophiala spp.) and other black pigmented fungi (Alternaria spp. and Cladosporium spp.), as well as tissues from uninfected sturgeon. The assay possessed high clinical specificity (100%) and clinical sensitivity (74%) in detecting V. botryosa DNA in splenic tissues from laboratory-infected sturgeon. Using V. botryosa genomic DNA as a template, the limit of detection was equivalent to 10 conidia, and the method was found suitable for the detection of fungal DNA in fresh and formalin-fixed tissues. In addition, the presence of non-target DNA from white sturgeon did not influence assay sensitivity. The developed qPCR assay is a sensitive, specific, and rapid diagnostic method for the detection and quantification of V. botryosa DNA from white sturgeon tissues.


Assuntos
Ascomicetos , Feoifomicose , Animais , Ascomicetos/genética , Peixes , Feoifomicose/veterinária , Reação em Cadeia da Polimerase/veterinária
15.
Trop Anim Health Prod ; 53(2): 322, 2021 May 14.
Artigo em Inglês | MEDLINE | ID: mdl-33988782

RESUMO

Bovine tuberculosis is an economically important disease with very high zoonotic potential. Single intradermal cervical tuberculin test (SICT) is considered a gold standard assay for the diagnosis of bovine tuberculosis. However, bovines especially buffaloes may produce a false negative result when the animal becomes cell-mediated immune (CMI) anergic in the advanced stage of the disease. In the present study, ELISA and PCR assays were successfully demonstrated to be useful in diagnosing tuberculosis especially in the CMI anergic buffaloes infected with Mycobacterium bovis. ELISA and PCR assays are able to detect 8.94% and 8.13%, respectively, more animals as positive in comparison to standard SICT assay in a selected population of 123 buffaloes. The moderate agreement between SICT and ELISA (k: 0.528; 0.249-0.807), a substantial agreement between SICT and PCR (k: 0.648; 0.364-0.931), and high agreement between ELISA and PCR (k: 0.856; 0.697-1.0) highlight that ELISA and PCR, if used in parallel with SICT, will provide better sensitivity over single assay. Reduction of false negative reactors may help in minimizing the zoonotic threat from bovine tuberculosis especially in disease endemic region where human and livestock interface is quite high.


Assuntos
Doenças dos Bovinos , Mycobacterium bovis , Tuberculose Bovina , Tuberculose , Animais , Búfalos , Bovinos , Ensaio de Imunoadsorção Enzimática/veterinária , Reação em Cadeia da Polimerase/veterinária , Sensibilidade e Especificidade , Tuberculina , Teste Tuberculínico/veterinária , Tuberculose/diagnóstico , Tuberculose/veterinária , Tuberculose Bovina/diagnóstico
16.
Rev Bras Parasitol Vet ; 30(2): e000621, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-33978118

RESUMO

Molecular methods such as Copro-PCR stand out in the diagnosis of T. gondii, because they are highly sensitive and specific, and can distinguish T. gondii from other morphologically similar coccids. The purpose was the detection of Toxoplasma gondii copro-prevalence by polymerase chain reaction in 149 fecal samples from stray and domiciled cats, using three distinct markers (B5-B6, 18S and 529bp RE). Oocysts of T. gondii/H. hammondi were detected in 15.4% by parasitology fecal tests (PFT), and 4% of these oocysts were positively identified as T. gondii by Copro-PCR. The presence of T. gondii genetic material was detected in 16.1%, but 12% of the samples that tested positive by Copro-PCR were negative in PFT. Samples with discordant results were subjected to a new Copro-PCR with 18S marker and a 529, and of the 17 samples, 9 contained T. gondii genetic material. A comparison of the PFT and the molecular methods showed the latter was more sensitive, since it detected 22.1% while the PFT detected 15.4%. Demonstrating the high sensitivity and specificity of the Copro-PCR, particularly with the association of primers (k=0.809), but also confirms the importance of using molecular techniques in laboratories, since Copro-PCR was able to detect samples considered negative by PFT.


Assuntos
Doenças do Gato , Toxoplasma , Toxoplasmose Animal , Animais , Doenças do Gato/diagnóstico , Doenças do Gato/epidemiologia , Gatos , DNA de Protozoário , Fezes , Oocistos , Reação em Cadeia da Polimerase/veterinária , Toxoplasma/genética , Toxoplasmose Animal/diagnóstico , Toxoplasmose Animal/epidemiologia
17.
Rev Bras Parasitol Vet ; 30(2): e027920, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-33978119

RESUMO

The present study aimed to evaluate a methodology for active surveillance of visceral leishmaniasis by detecting Leishmania DNA in organs of wild road-killed animals from November 2016 to October 2018 in the North of Paraná, Brazil. The collection points of road-killed wild animals were georeferenced. The animals were autopsied and samples of bone marrow, lymph node, liver, spleen, and ear skin were collected. Genomic DNA was extracted and subjected to PCR for amplification of Leishmania spp. 18S, kinetoplastic DNA (kDNA), HSP70, and ITS1 genes, and DNA sequencing was performed. The primers used for the amplification of kDNA, ITS1, and HSP70 genes presented non-specific results. Of the 66 mammals collected from 24 different municipalities, one Southern Tamandua (Tamandua tetradactyla) presented DNA of Leishmania spp. in lymph nodes by 18S PCR. DNA sequencing confirmed the results of the subgenus, Viannia, identification. We suggest using the methodology showed in the present study in the active and early surveillance of visceral leishmaniasis in a non-endemic area.


Assuntos
Leishmania , Leishmaniose Visceral , Leishmaniose , Animais , Brasil , Leishmania/genética , Leishmaniose/diagnóstico , Leishmaniose/epidemiologia , Leishmaniose/veterinária , Leishmaniose Visceral/diagnóstico , Leishmaniose Visceral/epidemiologia , Leishmaniose Visceral/veterinária , Mamíferos , Reação em Cadeia da Polimerase/veterinária
18.
Vet Rec ; 188(9): e143, 2021 05.
Artigo em Inglês | MEDLINE | ID: mdl-33960475

RESUMO

BACKGROUND: Bovine leptospirosis is mainly characterized by reproductive disorders. Contamination of the oocyte was previously demonstrated in vitro, resulting in some apparent damage. However, it is not clear whether it occurs under natural conditions. The present study aimed to characterize the presence of pathogenic Leptospira DNA in the ovarian follicles of non-pregnant cows. METHODS: Follicular fluid samples were collected from 65 animals and subjected to lipL32 PCR and secY sequencing. RESULTS: In total, seven of 65 (10.8%) were positive, indicating a possible early infection of the oocyte. Moreover, secY sequencing identified L. interrogans and L. santarosai, both very closely related to bovine strains from the Sejroe serogroup (Hardjoprjitno and Guaricura). We demonstrated that ovarian follicles can also be infected. CONCLUSIONS: It was hypothesised that ovarian infection can contribute to embryonic death, causing reproductive failure and estrus repetition. In the present study, we show that the organism identified in the follicle is closely related to one that is known to be associated with reproductive disorders.


Assuntos
Doenças dos Bovinos/microbiologia , DNA Bacteriano/isolamento & purificação , Líquido Folicular/microbiologia , Leptospira/genética , Leptospirose/veterinária , Animais , Bovinos , Feminino , Leptospirose/microbiologia , Reação em Cadeia da Polimerase/veterinária , Sorogrupo
19.
BMC Vet Res ; 17(1): 170, 2021 Apr 17.
Artigo em Inglês | MEDLINE | ID: mdl-33865378

RESUMO

BACKGROUND: Mycoplasma bovis is an important pathogen of cattle worldwide. Many different clinical manifestations of infection can occur, including respiratory disease, arthritis, and mastitis, causing heavy losses to beef and dairy industries. Because Mycoplasma species are slow-growing and fastidious, traditional identification methods are not cost- or time-effective, and improved methods are sought to streamline laboratory processes. High-resolution melting PCR (HRM-PCR) and matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) are 2 relatively recent tools that are rapid and inexpensive to use; we tested 9 isolates of M. bovis using both assays. The HRM-PCR assay used universal mycoplasma primers for the 16S-23S intergenic spacer region (IGSR). RESULTS: The resulting melting profiles of the field isolates were indistinguishable from the reference strain, indicating accurate identification. For the MALDI-TOF MS, each M. bovis isolate was accurately identified. Mycoplasma arginini and Mycoplasma alkalescens isolates did not identify as M. bovis when tested by either assay. CONCLUSIONS: Our work shows that either assay could be used to identify unknown M. bovis isolates. For future work, the MALDI-TOF MS library should be expanded to include more mycoplasmas, and the HRM-PCR assay should be tested on additional mycoplasmas to ensure that the melting profiles are sufficiently distinctive.


Assuntos
Mycoplasma bovis/isolamento & purificação , Reação em Cadeia da Polimerase/veterinária , Espectrometria de Massas por Ionização e Dessorção a Laser Assistida por Matriz/veterinária , Animais , Proteínas de Bactérias/análise , Mycoplasma , Mycoplasma bovis/química , Mycoplasma bovis/genética , Reação em Cadeia da Polimerase/métodos , RNA Ribossômico 16S/genética , RNA Ribossômico 23S/genética , Espectrometria de Massas por Ionização e Dessorção a Laser Assistida por Matriz/métodos
20.
Zoonoses Public Health ; 68(5): 544-548, 2021 08.
Artigo em Inglês | MEDLINE | ID: mdl-33822468

RESUMO

Blastocystis is a common protist colonizing the gastrointestinal tract of humans and various animals. The first subtyping of Blastocystis isolates in pigs and humans in Serbia revealed unusual avian-specific subtype ST6 in humans. In total, 48 pig faecal specimens collected on seven pig farms and 50 human faecal specimens positive to Blastocystis by microscopic examination were selected for the study. Eleven randomly selected PCR-positive pig samples and 10 samples from human patients (with gastrointestinal complaints) were subjected to SSU rDNA sequencing. Three subtypes were identified (ST3, ST5 and ST6) by phylogenetic analysis. ST5 was found in all pig samples; while in human samples, we detected ST3 and ST6. The latter subtype is relatively uncommon in Europe and highly adapted to avian hosts; therefore, the possibility of sporadic zoonotic transmission to human patients should not be ignored.


Assuntos
Infecções por Blastocystis/veterinária , Blastocystis/classificação , Doenças dos Suínos/parasitologia , Zoonoses , Animais , Blastocystis/isolamento & purificação , Infecções por Blastocystis/epidemiologia , Infecções por Blastocystis/parasitologia , Humanos , Filogenia , Reação em Cadeia da Polimerase/veterinária , Sérvia/epidemiologia , Suínos
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