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1.
Benef Microbes ; 11(5): 477-488, 2020 Sep 01.
Artigo em Inglês | MEDLINE | ID: mdl-32877228

RESUMO

Neonatal calf diarrhoea is one of the challenges faced by intensive farming, and probiotics are considered a promising approach to improve calves' health. The objective of this study was to evaluate the effect of potential probiotic lactobacilli on new-born dairy calves' growth, diarrhoea incidence, faecal score, cytokine expression in blood cells, immunoglobulin A (IgA) levels in plasma and faeces, and pathogen abundance in faeces. Two in vivo assays were conducted at the same farm in two annual calving seasons. Treated calves received one daily dose of the selected lactobacilli (Lactobacillus reuteri TP1.3B or Lactobacillus johnsonii TP1.6) for 10 consecutive days. A faecal score was recorded daily, average daily gain (ADG) was calculated, and blood and faeces samples were collected. Pathogen abundance was analysed by absolute qPCR in faeces using primers directed at Salmonella enterica, rotavirus, coronavirus, Cryptosporidium parvum and three Escherichia coli virulence genes (eae, clpG and Stx1). The faecal score was positively affected by the administration of both lactobacilli strains, and diarrhoea incidence was significantly lower in treated calves. No differences were found regarding ADG, cytokine expression, IgA levels and pathogen abundance. Our findings showed that oral administration of these strains could improve gastrointestinal health, but results could vary depending on the calving season, which may be related to pathogen seasonality and other environmental effects.


Assuntos
Doenças dos Bovinos/terapia , Diarreia , Lactobacillus johnsonii/metabolismo , Lactobacillus reuteri/metabolismo , Probióticos/uso terapêutico , Animais , Animais Recém-Nascidos , Bovinos , Doenças dos Bovinos/microbiologia , Doenças dos Bovinos/prevenção & controle , Infecções por Coronavirus/prevenção & controle , Infecções por Coronavirus/veterinária , Criptosporidiose/prevenção & controle , Citocinas/sangue , Indústria de Laticínios , Diarreia/prevenção & controle , Diarreia/terapia , Diarreia/veterinária , Infecções por Escherichia coli/prevenção & controle , Infecções por Escherichia coli/veterinária , Fezes/virologia , Trato Gastrointestinal/microbiologia , Imunoglobulina A/sangue , Infecções por Rotavirus/prevenção & controle , Infecções por Rotavirus/veterinária , Salmonelose Animal/prevenção & controle
2.
BMC Vet Res ; 16(1): 272, 2020 Aug 05.
Artigo em Inglês | MEDLINE | ID: mdl-32758221

RESUMO

BACKGROUND: Coronaviruses are notorious pathogens that cause diarrheic and respiratory diseases in humans and animals. Although the epidemiology and pathogenicity of coronaviruses have gained substantial attention, little is known about bovine coronavirus in cattle, which possesses a close relationship with human coronavirus. Bovine torovirus (BToV) is a newly identified relevant pathogen associated with cattle diarrhoea and respiratory diseases, and its epidemiology in the Chinese cattle industry remains unknown. RESULTS: In this study, a total of 461 diarrhoeic faecal samples were collected from 38 different farms in three intensive cattle farming regions and analysed. Our results demonstrated that BToV is present in China, with a low prevalence rate of 1.74% (8/461). The full-length spike genes were further cloned from eight clinical samples (five farms in Henan Province). Phylogenetic analysis showed that two different subclades of BToV strains are circulating in China. Meanwhile, the three BToV strains identified from dairy calves, 18,307, 2YY and 5YY, all contained the amino acid variants R614Q, I801T, N841S and Q885E. CONCLUSIONS: This is the first report to confirm the presence of BToV in beef and dairy calves in China with diarrhea, which extend our understanding of the epidemiology of BToVs worldwide.


Assuntos
Doenças dos Bovinos/virologia , Infecções por Torovirus/veterinária , Torovirus/isolamento & purificação , Animais , Bovinos , Doenças dos Bovinos/epidemiologia , China/epidemiologia , Diarreia/epidemiologia , Diarreia/veterinária , Diarreia/virologia , Fezes/virologia , Filogenia , RNA Viral , Análise de Sequência de DNA , Torovirus/genética , Infecções por Torovirus/epidemiologia , Infecções por Torovirus/virologia , Proteínas Virais/genética
3.
Aust Vet J ; 98(9): 429-437, 2020 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-32643274

RESUMO

Bovine viral diarrhoea virus (BVDV) is an economically significant disease affecting the Australian cattle industry, with losses stemming from decreased production and reproductive performance and control costs. However, these losses can be difficult to appreciate, particularly in endemic regions. Overall, there is a variable but high herd-level seroprevalence in Australia. Despite a potentially high financial burden of the disease, the onus for control ultimately falls on producers and strategies employed will vary between regions. A cross-sectional study, using a postal survey, was conducted in 2013 to evaluate the BVDV knowledge, attitudes and management practices utilised by Australian cattle producers. A total of 192 producers participated in the study, and results indicate that knowledge and attitudes towards disease risk are variable and can be improved. Producer knowledge of how persistently infected (PI) animals are produced was higher than that of disease outcomes or transmission pathways. Implementation of biosecurity practices was limited, with approximately half of respondents employing quarantine procedures for introduced stock and only 2% indicating they would antigen test introduced stock for BVDV. Approximately a third (36%) of producers reported engaging in BVDV control, with the majority of these using vaccination strategies over deliberate exposure to a PI. Knowledge of and engagement with BVDV control was positively influenced by the producer relationships with veterinarians. Findings from this study suggest that building on education and delivering a consistent message among stakeholders would likely improve producer awareness and understanding in relation to BVDV and support decision making in BVDV management.


Assuntos
Doença das Mucosas por Vírus da Diarreia Viral Bovina , Doenças dos Bovinos , Vírus da Diarreia Viral Bovina , Animais , Atitude , Austrália , Bovinos , Estudos Transversais , Diarreia/veterinária , Estudos Soroepidemiológicos
4.
Arch Virol ; 165(10): 2323-2333, 2020 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-32715325

RESUMO

To investigate the epidemic characteristics of porcine epidemic diarrhea virus (PEDV), 135 clinical samples (including intestinal tissues and feces) were collected from diseased piglets during outbreaks of diarrhea from 2015 to 2019 on farms in Henan and Shanxi provinces of China where swine had been immunized with attenuated PEDV (CV777). A total of 86 clinical samples (86/135, 63.7%) were positive for PEDV by RT-PCR, and subsequently, the complete spike (S) and ORF3 genes of 32 PEDV samples were sequenced. Phylogenetic analysis showed that the 32 PEDV strains obtained in this study belonged to group 2 (pandemic variant strains) and had a close relationship to 17 Chinese strains after 2010, two South Korean strains (KNU-1305 and KNU-1807), three American strains (PC22A-P140.BI, USA/Colorado/2013, and USA/OK10240-6/2017) and a Mexican strain (PEDV/MEX/QRO/02/2017), but differed genetically from a South Korean strain (SM98), a European strain (Br1/87), a Chinese strain (LZC), and a vaccine strain (CV777). G2-a subgroup strains were the dominant pandemic variant strains circulating in Henan and Shanxi provinces of China. Furthermore, a cross-recombination event was identified in the S region of the SX/TY2/2017 strain, and the putative parental strains were the epidemic strains CH/GDGZ/2012 and CH/YZ1/2015, identified in China in 2012 and 2015, respectively. These results provide further information about PEDV evolution, which could improve our understanding of the circulation of PEDV in Henan and Shanxi provinces. This information will also be helpful for developing new strategies for prevention and control of variant strains.


Assuntos
Infecções por Coronavirus/veterinária , Diarreia/veterinária , Surtos de Doenças , Genoma Viral , Vírus da Diarreia Epidêmica Suína/genética , Glicoproteína da Espícula de Coronavírus/genética , Doenças dos Suínos/epidemiologia , Proteínas Virais/genética , Animais , China/epidemiologia , Infecções por Coronavirus/epidemiologia , Infecções por Coronavirus/transmissão , Infecções por Coronavirus/virologia , Diarreia/epidemiologia , Diarreia/virologia , Fazendas , Fezes/virologia , Variação Genética , Intestinos/virologia , Filogenia , Vírus da Diarreia Epidêmica Suína/classificação , Vírus da Diarreia Epidêmica Suína/isolamento & purificação , Recombinação Genética , Suínos/virologia , Doenças dos Suínos/transmissão , Doenças dos Suínos/virologia
5.
Anal Chim Acta ; 1125: 57-65, 2020 Aug 15.
Artigo em Inglês | MEDLINE | ID: mdl-32674781

RESUMO

Porcine epidemic diarrhea virus (PEDV), porcine deltacoronavirus (PDCoV), and swine acute diarrhea syndrome-coronavirus (SADS-CoV) are three emerging and re-emerging coronaviruses (CoVs). Symptoms caused by these three viruses are extremely similar, including acute diarrhea, vomiting and even death in piglets. To date, strict biosecurity is still the most effective disease prevention and control measures, and the early detection of pathogens is the most important link. Here, we developed a microfluidic-RT-LAMP chip detection system for the first time, which could detected PEDV, PDCoV and SADS-CoV simultaneously, and had advantages of rapid, simple, sensitive, high-throughput, and accurate at point-of-care settings. The lowest detection limits of the microfluidic-RT-LAMP chip method are 101 copies/µL, 102 copies/µL and 102 copies/µL for PEDV, PDCoV and SADS-CoV, respectively. The whole detection procedure can be finished rapidly in 40 min without any cross-reaction with other common swine viruses. A total of 173 clinical swine fecal samples characterized with diarrheal symptoms were used to evaluate the performance of the newly developed system, which presented good stabilities (C.V.s<5%) and specificities (100%), and possessed sensitivities of 92.24%, 92.19% and 91.23% for PEDV, PDCoV and SADS-CoV respectively. In summary, the established microfluidic-RT-LAMP chip detection system could satisfy the demanding in field diagnoses, which was suitable for promotion in remote areas due to its fast, portable and cost-effective characters.


Assuntos
Coronavirus/genética , Técnicas de Amplificação de Ácido Nucleico/métodos , RNA Viral/análise , Alphacoronavirus/genética , Alphacoronavirus/isolamento & purificação , Animais , Coronavirus/isolamento & purificação , Diarreia/diagnóstico , Diarreia/veterinária , Diarreia/virologia , Fezes/virologia , Dispositivos Lab-On-A-Chip , Limite de Detecção , Técnicas de Amplificação de Ácido Nucleico/instrumentação , Sistemas Automatizados de Assistência Junto ao Leito , Vírus da Diarreia Epidêmica Suína/genética , Vírus da Diarreia Epidêmica Suína/isolamento & purificação , RNA Viral/metabolismo , Reprodutibilidade dos Testes , Sensibilidade e Especificidade , Suínos
6.
BMC Vet Res ; 16(1): 264, 2020 Jul 29.
Artigo em Inglês | MEDLINE | ID: mdl-32727468

RESUMO

BACKGROUND: Neonatal diarrhea remains one of the main causes of morbi-mortality in dairy calves under artificial rearing. It is often caused by infectious agents of viral, bacterial, or parasitic origin. Cows vaccination and colostrum intake by calves during the first 6 h of life are critical strategies to prevent severe diarrhea but these are still insufficient. Here we report the field evaluation of a product based on IgY antibodies against group A rotavirus (RVA), coronavirus (CoV), enterotoxigenic Escherichia coli, and Salmonella sp. This product, named IgY DNT, has been designed as a complementary passive immunization strategy to prevent neonatal calf diarrhea. The quality of the product depends on the titers of specific IgY antibodies to each antigen evaluated by ELISA. In the case of the viral antigens, ELISA antibody (Ab) titers are correlated with protection against infection in calves experimentally challenged with RVA and CoV (Bok M, et al., Passive immunity to control bovine coronavirus diarrhea in a dairy herd in Argentina, 2017), (Vega C, et al., Vet Immunol Immunopathol, 142:156-69, 2011), (Vega C, et al., Res Vet Sci, 103:1-10, 2015). To evaluate the efficiency in dairy farms, thirty newborn Holstein calves were randomly assigned to IgY DNT or control groups and treatment initiated after colostrum intake and gut closure. Calves in the IgY DNT group received 20 g of the oral passive treatment in 2 L of milk twice a day during the first 2 weeks of life. Animals were followed until 3 weeks of age and diarrhea due to natural exposure to infectious agents was recorded during all the experimental time. RESULTS: Results demonstrate that the oral administration of IgY DNT during the first 2 weeks of life to newborn calves caused a delay in diarrhea onset and significantly reduced its severity and duration compared with untreated calves. Animals treated with IgY DNT showed a trend towards a delay in RVA infection with significantly shorter duration and virus shedding compared to control calves. CONCLUSIONS: This indicates that IgY DNT is an effective product to complement current preventive strategies against neonatal calf diarrhea in dairy farms. Furthermore, to our knowledge, this is the only biological product available for the prevention of virus-associated neonatal calf diarrhea.


Assuntos
Anticorpos Antibacterianos/uso terapêutico , Anticorpos Antivirais/uso terapêutico , Doenças dos Bovinos/terapia , Diarreia/veterinária , Imunoglobulinas/uso terapêutico , Imunoterapia , Animais , Animais Recém-Nascidos , Anticorpos Antiprotozoários , Bovinos , Doenças dos Bovinos/microbiologia , Doenças dos Bovinos/virologia , Criptosporidiose/prevenção & controle , Indústria de Laticínios , Diarreia/microbiologia , Diarreia/terapia , Diarreia/virologia , Imunização Passiva/métodos , Imunização Passiva/veterinária
7.
Virulence ; 11(1): 707-718, 2020 01 01.
Artigo em Inglês | MEDLINE | ID: covidwho-517705

RESUMO

With the outbreak of the recent severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) in 2019, coronaviruses have become a global research hotspot in the field of virology. Coronaviruses mainly cause respiratory and digestive tract diseases, several coronaviruses are responsible for porcine diarrhea, such as porcine epidemic diarrhea virus (PEDV), porcine deltacoronavirus (PDCoV), and emerging swine acute diarrhea syndrome coronavirus (SADS-CoV). Those viruses have caused huge economic losses and are considered as potential public health threats. Porcine torovirus (PToV) and coronaviruses, sharing similar genomic structure and replication strategy, belong to the same order Nidovirales. Here, we developed a multiplex TaqMan-probe-based real-time PCR for the simultaneous detection of PEDV, PDCoV, PToV, and SADS-CoV for the first time. Specific primers and TaqMan fluorescent probes were designed targeting the ORF1a region of PDEV, PToV, and SADS-CoV and the ORF1b region of PDCoV. The method showed high sensitivity and specificity, with a detection limit of 1 × 102 copies/µL for each pathogen. A total of 101 clinical swine samples with signs of diarrhea were analyzed using this method, and the result showed good consistency with conventional reverse transcription PCR (RT-PCR). This method improves the efficiency for surveillance of these emerging and reemerging swine enteric viruses and can help reduce economic losses to the pig industry, which also benefits animal and public health.


Assuntos
Doenças Transmissíveis Emergentes/veterinária , Infecções por Coronaviridae/veterinária , Coronaviridae/isolamento & purificação , Reação em Cadeia da Polimerase , Doenças dos Suínos/diagnóstico , Animais , Coinfecção/diagnóstico , Coinfecção/veterinária , Doenças Transmissíveis Emergentes/diagnóstico , Coronaviridae/genética , Infecções por Coronaviridae/diagnóstico , Diarreia/diagnóstico , Diarreia/veterinária , Fases de Leitura Aberta/genética , Reação em Cadeia da Polimerase/normas , RNA Viral/genética , Reprodutibilidade dos Testes , Sensibilidade e Especificidade , Suínos
8.
Virulence ; 11(1): 707-718, 2020 01 01.
Artigo em Inglês | MEDLINE | ID: mdl-32490723

RESUMO

With the outbreak of the recent severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) in 2019, coronaviruses have become a global research hotspot in the field of virology. Coronaviruses mainly cause respiratory and digestive tract diseases, several coronaviruses are responsible for porcine diarrhea, such as porcine epidemic diarrhea virus (PEDV), porcine deltacoronavirus (PDCoV), and emerging swine acute diarrhea syndrome coronavirus (SADS-CoV). Those viruses have caused huge economic losses and are considered as potential public health threats. Porcine torovirus (PToV) and coronaviruses, sharing similar genomic structure and replication strategy, belong to the same order Nidovirales. Here, we developed a multiplex TaqMan-probe-based real-time PCR for the simultaneous detection of PEDV, PDCoV, PToV, and SADS-CoV for the first time. Specific primers and TaqMan fluorescent probes were designed targeting the ORF1a region of PDEV, PToV, and SADS-CoV and the ORF1b region of PDCoV. The method showed high sensitivity and specificity, with a detection limit of 1 × 102 copies/µL for each pathogen. A total of 101 clinical swine samples with signs of diarrhea were analyzed using this method, and the result showed good consistency with conventional reverse transcription PCR (RT-PCR). This method improves the efficiency for surveillance of these emerging and reemerging swine enteric viruses and can help reduce economic losses to the pig industry, which also benefits animal and public health.


Assuntos
Doenças Transmissíveis Emergentes/veterinária , Infecções por Coronaviridae/veterinária , Coronaviridae/isolamento & purificação , Reação em Cadeia da Polimerase , Doenças dos Suínos/diagnóstico , Animais , Coinfecção/diagnóstico , Coinfecção/veterinária , Doenças Transmissíveis Emergentes/diagnóstico , Coronaviridae/genética , Infecções por Coronaviridae/diagnóstico , Diarreia/diagnóstico , Diarreia/veterinária , Fases de Leitura Aberta/genética , Reação em Cadeia da Polimerase/normas , RNA Viral/genética , Reprodutibilidade dos Testes , Sensibilidade e Especificidade , Suínos
9.
Emerg Infect Dis ; 26(7): 1610-1612, 2020 07.
Artigo em Inglês | MEDLINE | ID: mdl-32568058

RESUMO

We characterized novel coronaviruses detected in US bottlenose dolphins (BdCoVs) with diarrhea. These viruses are closely related to the other 2 known cetacean coronaviruses, Hong Kong BdCoV and beluga whale CoV. A deletion in the spike gene and insertions in the membrane gene and untranslated regions were found in US BdCoVs (unrelated to severe acute respiratory syndrome coronavirus 2).


Assuntos
Golfinho Nariz-de-Garrafa/virologia , Infecções por Coronavirus/veterinária , Diarreia/veterinária , Gammacoronavirus/classificação , Gammacoronavirus/genética , Animais , Infecções por Coronavirus/virologia , Diarreia/virologia , Gammacoronavirus/isolamento & purificação , Gammacoronavirus/fisiologia , Genes Virais , Genoma Viral , Mutação , Filogenia , Deleção de Sequência , Glicoproteína da Espícula de Coronavírus/genética , Proteínas da Matriz Viral/genética
10.
Trop Anim Health Prod ; 52(5): 2761-2768, 2020 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-32488696

RESUMO

Group A rotavirus (RVA) and bovine coronavirus (BCoV) are the two main viral enteropathogens associated with neonatal calf diarrhea. The aim of the present survey was to investigate the epidemiology and the role of RVA and BCoV in the presentation of dairy and beef calf diarrhea in Lerma Valley of Salta province, within the Northwest region of Argentina. Stool samples of calves with or without diarrhea younger than 2 months of age were collected from 19 dairy farms and 20 beef farms between the years 2014 and 2016. Stool samples were screened for RVA and BCoV detection by ELISA. Heminested multiplex RT-PCR was used for RVA typing and RT-PCR to confirm BCoV. Positive samples were submitted to sequencing analysis. Bovine RVA and BCoV were circulating in 63% (12/19) and 10.52% (2/19) of the dairy farms, respectively, where 9.5% (46/484) of the calves were positives to RVA and 0.4% (2/484) to BCoV. In beef herds, RVA was detected in 40% (8/20) of the farms and in 6.75% (21/311) of the calves, without positives cases of BCoV. Molecular analysis showed that in dairy farms, G6P[11] and G10P[11] were the prevalent RVA strains, while in beef farms, G10P[11] was the prevalent. The main finding was the detection for the first time of a G15P[11] causing diarrhea in beef calves of Argentina that represents a new alert to be consider for future vaccine updates. Analysis of detected BCoV showed that it is related to the other circulating strains of Argentina.


Assuntos
Doenças dos Bovinos/virologia , Coronavirus Bovino/isolamento & purificação , Diarreia/veterinária , Infecções por Rotavirus/veterinária , Rotavirus/isolamento & purificação , Animais , Animais Recém-Nascidos , Argentina , Bovinos , Coronavirus Bovino/genética , Diarreia/virologia , Ensaio de Imunoadsorção Enzimática/veterinária , Fezes/virologia , Genótipo , Rotavirus/genética , Infecções por Rotavirus/virologia
11.
J Vet Diagn Invest ; 32(4): 513-526, 2020 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-32484424

RESUMO

Bovine coronaviruses (BoCVs) have been found in respiratory tissues in cattle and frequently associated with bovine respiratory disease (BRD); however, pathogenesis studies in calves are limited. To characterize the pathogenesis and pathogenicity of BoCV isolates, we used 5 different BoCV strains to inoculate colostrum-deprived calves, ~ 2-5 wk of age. Later, to determine if dual viral infection would potentiate pathogenicity of BoCV, calves were inoculated with BoCV alone, bovine viral diarrhea virus (BVDV) alone, or a series of dual-infection (BVDV-BoCV) schemes. A negative control group was included in all studies. Clinical signs and body temperature were monitored during the study and samples collected for lymphocyte counts, virus isolation, and serology. During autopsy, gross lesions were recorded and fixed tissues collected for histopathology and immunohistochemistry; fresh tissues were collected for virus isolation. Results suggest increased pathogenicity for isolate BoCV OK 1776. Increased body temperature was found in all virus-inoculated groups. Lung lesions were present in calves in all dual-infection groups; however, lesions were most pronounced in calves inoculated with BVDV followed by BoCV inoculation 6 d later. Lung lesions were consistent with mild-to-moderate interstitial pneumonia, and immunohistochemistry confirmed the presence of BoCV antigen. Our studies demonstrated that BVDV-BoCV dual infection may play an important role in BRD pathogenesis, and timing between infections seems critical to the severity of lesions.


Assuntos
Anticorpos Antivirais/sangue , Doença das Mucosas por Vírus da Diarreia Viral Bovina/virologia , Coronavirus Bovino/isolamento & purificação , Vírus da Diarreia Viral Bovina Tipo 1/isolamento & purificação , Doenças Respiratórias/veterinária , Animais , Doença das Mucosas por Vírus da Diarreia Viral Bovina/patologia , Bovinos , Colostro , Diarreia/veterinária , Vírus da Diarreia Viral Bovina/imunologia , Feminino , Gravidez , Doenças Respiratórias/patologia , Doenças Respiratórias/virologia
12.
Mol Cell Probes ; 53: 101618, 2020 10.
Artigo em Inglês | MEDLINE | ID: mdl-32534013

RESUMO

Viral canine diarrhea has high morbidity and mortality and is prevalent worldwide, resulting in severe economic and spiritual losses to pet owners. However, diarrhea pathogens have similar clinical symptoms and are difficult to diagnose clinically. Thus, fast and accurate diagnostic methods are of great significance for prevention and accurate treatment. In this study, we developed a one-step multiplex TaqMan probe-based real-time PCR for the differential diagnosis of four viruses causing canine diarrhea including, CPV (Canine Parvovirus), CCoV (Canine Coronavirus), CAstV (Canine Astrovirus), and CaKoV (Canine Kobuviruses). The limit of detection was up to 102 copies/µL and performed well with high sensitivity and specificity. This assay was optimized and used to identify possible antagonistic relationships between viruses. From this, artificial pre-experiments were performed for mixed infections, and a total of 82 canine diarrhea field samples were collected from different animal hospitals in Zhejiang, China to assess the method. The virus prevalence was significantly higher than what previously reported based on RT-PCR (Reverse Transcription-Polymerase Chain Reaction). Taken together, these results suggest that the method can be used as a preferred tool for monitoring laboratory epidemics, timely prevention, and effective monitoring of disease progression.


Assuntos
Sondas de DNA , Diarreia/veterinária , Doenças do Cão/virologia , Reação em Cadeia da Polimerase em Tempo Real/veterinária , Animais , Avastrovirus , Coronavirus Canino , Diarreia/diagnóstico , Diarreia/virologia , Doenças do Cão/diagnóstico , Cães , Kobuvirus , Parvovirus Canino , Reação em Cadeia da Polimerase em Tempo Real/métodos , Reprodutibilidade dos Testes , Sensibilidade e Especificidade
13.
PLoS One ; 15(5): e0232890, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32392237

RESUMO

Diarrhea caused by Escherichia coli in calves is an important problem in terms of survivability, productivity and treatment costs. In this study, 88 of 150 diarrheic animals tested positive for E. coli. Of these, 54 samples had mixed infection with other bacterial and/or parasitic agents. There are several diarrheagenic E. coli pathotypes including enteropathogenic E. coli (EPEC), Shiga-toxin producing E. coli (STEC), enterotoxigenic E. coli (ETEC) and necrotoxigenic E. coli (NTEC). Molecular detection of virulence factors Stx2, Cdt3, Eae, CNF2, F5, Hly, Stx1, and ST revealed their presence at 39.7, 27.2, 19.3, 15.9, 13.6, 9.0, 3.4, and 3.4 percent, respectively. As many as 13.6% of the isolates lacked virulence genes and none of the isolate had LT or CNF1 toxin gene. The odds of isolating ETEC from male calves was 3.6 times (95% CI: 1.1, 12.4; P value = 0.042) that of female calves, whereas the odds of isolating NTEC from male calves was 72.9% lower (95% CI: 91.3% lower, 15.7% lower; P value = 0.024) than that in females. The odds of isolating STEC in winter was 3.3 times (95% CI: 1.1, 10.3; P value = 0.037) that of spring. Antibiograms showed 48 (54.5%) of the isolates to be multi-drug resistant. The percent resistance to tetracycline, streptomycin, ampicillin, and trimethoprim-sulfamethoxazole was 79.5, 67.0, 54.5, and 43.0, respectively. Ceftazidime (14.8%), amoxicillin-clavulanic acid (13.6%) and aztreonam (11.3%) showed the lowest resistance, and none of the isolates was resistant to imipenem. The results of this study can help improve our understanding of the epidemiological aspects of E. coli infection and to devise strategies for protection against it. The prevalence of E. coli pathotypes can help potential buyers of calves to avoid infected premises. The antibiograms in this study emphasizes the risks associated with the random use of antibiotics.


Assuntos
Doenças dos Bovinos/epidemiologia , Doenças dos Bovinos/microbiologia , Diarreia/veterinária , Infecções por Escherichia coli/veterinária , Criação de Animais Domésticos , Animais , Búfalos , Bovinos , Diarreia/epidemiologia , Diarreia/microbiologia , Farmacorresistência Bacteriana Múltipla/genética , Escherichia coli/efeitos dos fármacos , Escherichia coli/genética , Escherichia coli/isolamento & purificação , Infecções por Escherichia coli/epidemiologia , Proteínas de Escherichia coli/genética , Feminino , Masculino , Testes de Sensibilidade Microbiana , Fatores de Risco , Estações do Ano , Fatores de Virulência/genética
14.
Arch Virol ; 165(7): 1577-1583, 2020 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-32388597

RESUMO

Bovine torovirus (BToV) is a diarrhea-causing pathogen. In this study, 92 diarrheic fecal samples from five farms in four provinces in China were collected and tested for BToV using a RT-PCR assay, and 21.73% samples were found to be BToV positive. Moreover, two complete BToV genome sequences (MN073058 and MN073059) were obtained from the clinical samples, which were 28,297 and 28,301 nucleotides in length, respectively. Sequence analysis showed that the two isolates shared 10 identical amino acid mutations in the S protein compared to the complete S sequences of BToV available in the GenBank database. In addition, seven consecutive amino acid mutations were found from aa 1,486 to 1,492 in the S protein of isolate MN073058. Moreover, the two isolates shared one identical amino acid mutation in the receptor binding sites of the HE protein. To the best of our knowledge, this is the first report on the epidemic and genomic characterization of BToV in China, which is helpful for further understanding the genetic evolution of BToV.


Assuntos
Doenças dos Bovinos/virologia , Diarreia/veterinária , Infecções por Torovirus/veterinária , Torovirus/isolamento & purificação , Animais , Bovinos , Diarreia/virologia , Fezes/virologia , Genoma Viral , Genômica , Filogenia , Torovirus/classificação , Torovirus/genética , Infecções por Torovirus/virologia , Proteínas Virais/genética
15.
PLoS One ; 15(4): e0231723, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32343704

RESUMO

Understanding the immune responses against Porcine epidemic diarrhea virus (PEDV) is important to prevent infection and to design control strategies. We evaluated both systemic and mucosal immune responses to PEDV in pigs and assessed if prior exposure to virus protects against re-infection. Three-week-old pigs were infected with PEDV and immune response in blood, intestine, and mesenteric lymph node (MLN) was evaluated. At 30 dpi, virus exposed pigs were challenged with a field isolate of PEDV and immune response at 5 d post challenge was evaluated. We found that PEDV RNA persists in the intestine even after fecal shedding of the virus was stopped at 28 dpi and pigs previously exposed to PEDV are protected from virus shedding after re-infection. PEDV infection induced both humoral and cell mediated immune response with an increase in PEDV specific IgA and IgG antibodies in intestine and serum. Flow cytometry analysis showed a significantly higher frequency of B cells and lower frequency of T cells at 4 dpi. The frequency of CD4/CD8 double positive (DP) memory T cells was significantly increased in the MLN of challenged animals. These studies may provide further insights into understanding the mucosal immune response to PEDV and its role in protection against disease.


Assuntos
Anticorpos Antivirais/análise , Infecções por Coronavirus/imunologia , Diarreia/imunologia , Vírus da Diarreia Epidêmica Suína/imunologia , Animais , Anticorpos Antivirais/imunologia , Anticorpos Antivirais/metabolismo , Linfócitos B/imunologia , Infecções por Coronavirus/sangue , Infecções por Coronavirus/veterinária , Infecções por Coronavirus/virologia , Diarreia/sangue , Diarreia/veterinária , Diarreia/virologia , Resistência à Doença/imunologia , Fezes/microbiologia , Imunidade Celular , Imunidade Humoral , Imunidade nas Mucosas , Imunoglobulina A/análise , Imunoglobulina A/imunologia , Imunoglobulina A/metabolismo , Imunoglobulina G/análise , Imunoglobulina G/imunologia , Imunoglobulina G/metabolismo , Mucosa Intestinal/imunologia , Mucosa Intestinal/metabolismo , Vírus da Diarreia Epidêmica Suína/genética , Vírus da Diarreia Epidêmica Suína/isolamento & purificação , RNA Viral/isolamento & purificação , Suínos , Linfócitos T/imunologia , Eliminação de Partículas Virais
16.
PLoS One ; 15(4): e0230708, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32271786

RESUMO

Neonatal calf diarrhea (NCD) is a major problem to calf health worldwide, in terms of both morbidity and mortality. A five-point ordinal scale clinical assessment scoring (CAS) chart was utilized to assess calves suffering from NCD-related clinical abnormalities (acidosis and dehydration) on commercial farms. The objective of this research was to determine the predictive capability of this CAS chart against gold standard blood gas parameters, designed to assist farmers in the accurate assessment of the clinical consequences of NCD. A total of 443 diarrheic and non-diarrheic calves were enrolled in the study. The CAS chart rated a calf's health from no clinical signs to varying degrees of clinical severity on a 0 (clinically normal) to 4 (grave) scale, based on clinical indicators including calf demeanour, ear position, mobility, suckle reflex, desire-to-feed, and enophthalmos. Blood gas analysis was conducted for individual calves, consisting of pH, base excess, Na+, K+, Ca2+, Cl-, glucose, total hemoglobin, bicarbonate, anion gap, and strong ion difference. Statistical evaluation was performed by comparison of the CAS score with blood gas profiles using ordinal logistic regression and a non-parametric estimation of the Receiver Operating Characteristics (ROC). The ROC analysis indicated that the CAS chart had acceptable specificity (>95%) with low sensitivity (<60%) in differentiating clinically normal from acidotic/dehydrated cases. Assessment of individual severity classes indicated that the chart can predict and differentiate both clinically normal and advanced cases from the other severity classes (peak estimations >80%) but had reduced accuracy in differentiating mild and moderate cases (peak estimations >50%). The chart, as presented, provides a simple tool to differentiate clinically normal from calves suffering the consequences of diarrhea, but fails to accurately differentiate severity for NCD related acidosis and dehydration. Further efforts are required to enhance the sensitivity and differential diagnostic value of this type of chart.


Assuntos
Animais Recém-Nascidos , Doenças dos Bovinos/diagnóstico , Técnicas e Procedimentos Diagnósticos/veterinária , Diarreia/diagnóstico , Design de Software , Acidose/sangue , Acidose/diagnóstico , Acidose/veterinária , Animais , Gasometria/normas , Gasometria/veterinária , Bovinos , Doenças dos Bovinos/sangue , Doenças dos Bovinos/patologia , Desidratação/sangue , Desidratação/diagnóstico , Desidratação/veterinária , Técnicas e Procedimentos Diagnósticos/normas , Diarreia/sangue , Diarreia/patologia , Diarreia/veterinária , Feminino , Masculino , Valor Preditivo dos Testes , Prognóstico , Projetos de Pesquisa , Índice de Gravidade de Doença
17.
J Vet Sci ; 21(2): e17, 2020 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-32233127

RESUMO

Dehydration, electrolyte disturbance, and acid-base imbalance are the most significant consequences of diarrhea in calves. We aimed to determine blood gas, hematological, electrolyte, and biochemical values and investigate the relationship between the physical status and blood parameters in Korean native calves (KNCs) with diarrhea. One hundred eighty KNCs with diarrhea (age < 75 days) were investigated. Blood samples were collected from the external jugular vein and analyzed using a portable clinical blood gas analyzer. The measured parameters were statistically compared according to the status of physical activity, dehydration, or prognosis. The mean values of parameters in the Calves with diarrhea showed metabolic acidosis, hyponatremia, and azotemia. The mean values of potassium, chloride, hematocrit, and hemoglobin were in the upper limit of their reference ranges. More than 75% of the calves had metabolic acidosis caused by bicarbonate loss, and 63.6% had high blood urea nitrogen (BUN) values. Moreover, BUN showed the highest correlation with the physical activity status and dehydration. pH, base excess of the extracellular fluid (BE), anion gap, potassium, hematocrit, bicarbonate, and hemoglobin were closely correlated with physical deterioration and dehydration (p < 0.001). BUN, pH, BE, and anion gap were closely correlated with physical deterioration and dehydration. These correlations between clinical symptoms and blood gas parameters can be clinically relevant in predicting the status of parameters according to clinical symptoms.


Assuntos
Doenças dos Bovinos , Diarreia/veterinária , Animais , Animais Recém-Nascidos , Análise Química do Sangue/veterinária , Bovinos , Doenças dos Bovinos/sangue , Doenças dos Bovinos/fisiopatologia , Diarreia/sangue , Diarreia/fisiopatologia , Feminino , Testes Hematológicos/veterinária , Masculino , República da Coreia
18.
J Vet Sci ; 21(2): e25, 2020 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-32233133

RESUMO

Porcine epidemic diarrhea virus (PEDV) emerged into Canada in January 2014, primarily affecting sow herds. Subsequent epidemiological analyses suggested contaminated feed was the most likely transmission pathway. The primary objective of this study was to describe general biosecurity and management practices implemented in PEDV-positive sow herds and matched control herds at the time the virus emerged. The secondary objective was to determine if any of these general biosecurity and farm management practices were important in explaining PEDV infection status from January 22, 2014 to March 1, 2014. A case herd was defined as a swine herd with clinical signs and a positive test result for PEDV. A questionnaire was used to a gather 30-day history of herd management practices, animal movements on/off site, feed management practices, semen deliveries and biosecurity practices for case (n = 8) and control (n = 12) herds, primarily located in Ontario. Data was analyzed using descriptive statistics and random forests (RFs). Case herds were larger in size than control herds. Case herds had more animal movements and non-staff movements onto the site. Also, case herds had higher quantities of pigs delivered, feed deliveries and semen deliveries on-site. The biosecurity practices of case herds were considered more rigorous based on herd management, feed deliveries, transportation and truck driver practices than control herds. The RF model found that the most important variables for predicting herd status were related to herd size and feed management variables. Nonetheless, predictive accuracy of the final RF model was 72%.


Assuntos
Criação de Animais Domésticos/métodos , Infecções por Coronavirus/veterinária , Diarreia/veterinária , Enterite/veterinária , Doenças dos Suínos/prevenção & controle , Animais , Infecções por Coronavirus/prevenção & controle , Infecções por Coronavirus/virologia , Diarreia/prevenção & controle , Diarreia/virologia , Enterite/prevenção & controle , Enterite/virologia , Fazendas , Ontário , Vírus da Diarreia Epidêmica Suína/fisiologia , Suínos , Doenças dos Suínos/virologia
19.
Trop Anim Health Prod ; 52(5): 2743-2747, 2020 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-32279241

RESUMO

Porcine epidemic diarrhea (PED) is a devastating enteric disease causing economic losses in many countries including the Philippines. To control PED, apart from oral administration of minced intestinal materials, there are still no effective control methods. The ability of porcine epidemic diarrhea vaccine RNA particle platform (PED-RP) to induce antibody in colostrum and milk samples was investigated in two pig herds with a differing PED status in the Philippines. Herd-A was naïve but herd-B was endemically infected with PED. Sera, colostrum, and milk samples were collected prior to and following vaccination, and assayed for the presence of antibody by viral neutralization (VN) and IgG and IgA levels by ELISA spike protein. The results from both herds, compared to the non-vaccinated control group, demonstrated significantly increased VN titers and IgG and IgA levels in colostrum and milk samples of sows at 0, 7, 14, and 21 days post parturition. Additionally, piglets from vaccinated sows had VN titers, and IgG and IgA levels are significantly higher than those from non-vaccinated sows. In conclusion, the results of the study demonstrate that PED-RP can be used to induce a satisfactory antibody response in colostrum and milk, as measured by VN titers and IgG and IgA levels.


Assuntos
Anticorpos Antivirais/sangue , Infecções por Coronavirus/veterinária , Vírus da Diarreia Epidêmica Suína/imunologia , Doenças dos Suínos/prevenção & controle , Vacinas Virais/imunologia , Animais , Colostro/imunologia , Infecções por Coronavirus/prevenção & controle , Diarreia/veterinária , Ensaio de Imunoadsorção Enzimática/veterinária , Feminino , Leite/química , Filipinas , Gravidez , RNA , Distribuição Aleatória , Suínos , Vacinação/veterinária , Vacinas de Produtos Inativados/imunologia
20.
J Anim Sci ; 98(5)2020 May 01.
Artigo em Inglês | MEDLINE | ID: mdl-32253427

RESUMO

Fifty-six piglets (6.26 ± 0.64 kg BW) were weaned at 21 d and randomly assigned to one of the eight dietary treatments with seven replicate pens for a 14-d experimental period. The eight experimental diets were prepared via a 2 × 4 factorial arrangement with citric acid (CA; 0% and 0.3%) and dietary electrolyte balance (dEB, Na + K - Cl mEq/kg of the diet; -50, 100, 250, and 400 mEq/kg). Varying dEB values were obtained by altering the contents of calcium chloride and sodium bicarbonate. An interaction (P < 0.05) between dEB and CA in diarrhea score and the number of goblet cell in jejunum were observed. Ileum pH significantly decreased in weaned piglets fed 250 mEq/kg dEB diet compared with those fed -50 and 400 mEq/kg dEB diets (P < 0.05). Supplementation of 0.3% CA decreased the number of goblet cell in the ileal crypt (P < 0.05) and the relative mRNA expression of cystic fibrosis transmembrane conductance regulator, tumor necrosis factor-α, interferon-γ (IFN-γ), interleukin-1ß (IL-1ß), interleukin-10 (IL-10), zona occludens-1, and Claudin-1 (P < 0.05). Increasing dEB values increased the number of goblet cells in the jejunal crypt (P < 0.05). A 250-mEq/kg dEB diet decreased the relative mRNA expression of IFN-γ, IL-1ß, and IL-10 (P < 0.05) than 100-mEq/kg dEB diet. The interaction between dEB and CA on the relative abundances of Cyanobacteria and Saccharibacteria was observed (P < 0.05). Supplementation of 0.3% CA increased relative abundances of and Streptococcus hyointestinalis. Piglets fed 250-mEq/kg diet increased relative abundances of Firmicutes and Lactobacillus rennini, and decreased the relative abundance of Proteobacteria, Veillonella, Actinobacillus minor, and Escherichia-Shigella.In conclusion, supplementation of 0.3% CA resulted in differential expression of inflammatory cytokines, ion transporters, and tight junction proteins, and changes in the microbial community composition. A 250-mEq/kg dEB diet reduced gastrointestinal pH and promoted the enrichment of beneficial microbes in the gut microbiota, thereby suppressing inflammation and harmful bacteria. However, the addition of CA to diets with different dEB values did not promote intestinal function in weaned piglets.


Assuntos
Ácido Cítrico/farmacologia , Diarreia/veterinária , Suplementos Nutricionais/análise , Microbioma Gastrointestinal , Doenças dos Suínos/metabolismo , Equilíbrio Hidroeletrolítico , Animais , Citocinas/metabolismo , Diarreia/metabolismo , Diarreia/microbiologia , Dieta/veterinária , Intestinos/microbiologia , Intestinos/fisiologia , Masculino , Distribuição Aleatória , Suínos , Doenças dos Suínos/microbiologia , Desmame
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