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1.
Pol J Vet Sci ; 22(4): 793-797, 2019 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-31867930

RESUMO

Salmonellosis is a public health concern worldwide and also causes huge losses to the piggery industry. A total of 457 fecal samples were collected from organized and unorganized farms including indigenous and crossbreed piglets of North East India. Salmonella isolates were serotyped, screened for their virulence genes, characterized for drug resistance pattern and representative isolates were cloned and sequenced for their partial length enterotoxin (stn) gene. A total of 8.31% Salmonella were identified with higher prevalence observed in unorganized compared to organized farms and higher detection level in cross breed compared to indigenous piglets. Salmonella typhimurium (65.78%) was found to be the predominant serovar and irrespective of serovars high number of isolates (68.4%) harboured enterotoxin gene. The isolates were multidrug resistant showing highest resistance against cefalexin (77.31%). Sequence analysis of stn gene showed two isolates having diverse sequence compared to other isolates. Our study revealed the significance of Salmonella as important pathogen with zoonotic potential between porcine and human populations. This is probably the first systematic study of Salmonella species associated with piglet diarrhea in India.


Assuntos
Diarreia/veterinária , Salmonelose Animal/microbiologia , Salmonella/classificação , Doenças dos Suínos/microbiologia , Animais , Antibacterianos/farmacologia , Diarreia/epidemiologia , Diarreia/microbiologia , Farmacorresistência Bacteriana , Fezes/microbiologia , Índia/epidemiologia , Salmonella/efeitos dos fármacos , Salmonella/genética , Salmonelose Animal/epidemiologia , Suínos
2.
Vet Microbiol ; 239: 108459, 2019 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-31767067

RESUMO

Helicobacter suis is a fastidious, Gram negative bacterium that colonizes the stomach of pigs and non-human primates. It has also been associated with gastric disease in humans. A combined agar and broth dilution method was used to analyze the activity of 15 antimicrobial agents against 20 and 15 H. suis isolates obtained from pigs and macaques, respectively. After 48 h microaerobic incubation, minimal inhibitory concentrations (MICs) were determined by software-assisted calculation of bacterial growth as determined by quantitative real-time PCR. A monomodal distribution of MICs was seen for ß-lactam antibiotics, macrolides, gentamicin, neomycin, doxycycline, metronidazole, and rifampicin. Presence of a bimodal distribution of MICs indicated that 2 porcine isolates did not belong to the wild type population (WTP) for fluoroquinolones. This was also the case for 1 porcine isolate for tetracycline, 1 porcine and 2 primate isolates for lincomycin, and 1 primate isolate for spectinomycin. Single nucleotide polymorphisms (SNPs) were present in the gyrA gene of the isolates not belonging to the WTP for fluoroquinolones and in ribosomal protein encoding genes of the isolates not belonging to the WTP for tetracycline and spectinomycin. MICs of ampicillin, tetracycline and doxycycline were higher for porcine H. suis isolates compared to primate isolates and in these porcine isolates SNPs were detected in genes encoding penicillin binding and ribosomal proteins. This study indicates that acquired resistance occasionally occurs in H. suis isolates and that zoonotically important porcine isolates may be intrinsically less susceptible to ß-lactam antibiotics and tetracyclines than primate isolates.


Assuntos
Antibacterianos/farmacologia , Infecções por Helicobacter/microbiologia , Helicobacter heilmannii/efeitos dos fármacos , Doenças dos Macacos/microbiologia , Doenças dos Suínos/microbiologia , Animais , DNA Girase/genética , Farmacorresistência Bacteriana , Helicobacter heilmannii/isolamento & purificação , Macaca/microbiologia , Testes de Sensibilidade Microbiana , Polimorfismo de Nucleotídeo Único , Suínos/microbiologia
3.
Vet Microbiol ; 239: 108457, 2019 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-31767083

RESUMO

Pork is one of the most common vehicles of non-typhoid foodborne Salmonella, with the slaughterhouse representing a key point for the infection of pigs and carcass contamination. By comparing matching samples taken from animals at the dirty (skin) and clean (inner and outer carcass surface) areas of the slaughterline, this study aimed to assess potential Salmonella contamination routes of pig carcasses within a Portuguese abattoir. Forty-four Salmonella isolates were retrieved from 120 pigs, and further characterized through pheno and genotypical methods. Most frequent serotypes found were Salmonella 4, [5],12:i:- (47.7%), Salmonella Rissen (40.9%) and Salmonella Derby (11.4%). Isolates were most commonly collected from the skin of pigs sampled at the dirty area (59.1%), followed by the inner (38.1%) and outer (9.1%) carcass surface sampled at the clean area. Most isolates (79.5%) were considered to be multidrug resistant and all harbored the virulence associated genes invA, invH, sopB, stn, slyA, phoP, phoQ and agfA. PFGE analysis revealed that most bacterial isolates belonging to the same serotype, recovered from animals from different farms, and slaughtered at separate days were genetically undistinguishable. Furthermore, our findings suggest that Salmonella Rissen might have an increased ability to endure on the slaughterhouse environment when compared with the other serotypes. Concluding, this study shows that the slaughterhouse may be a key point for the dissemination of resistant and virulent Salmonella strains, which stresses the importance of the implementation of good hygiene practices at the slaughterhouse and of the application of corrective measures to avoid cross-contamination.


Assuntos
Matadouros , Microbiologia de Alimentos , Salmonelose Animal/microbiologia , Salmonella/classificação , Salmonella/genética , Doenças dos Suínos/microbiologia , Animais , Resistência a Múltiplos Medicamentos , Genótipo , Carne/microbiologia , Fenótipo , Portugal , Sorotipagem , Suínos , Tempo , Fatores de Virulência/genética
4.
Vet Microbiol ; 239: 108448, 2019 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-31767090

RESUMO

This study was designed to detect Mycoplasma hyorhinis and M. hyosynoviae in oral fluids and determine their correlation with lameness scores in pigs. Thirty-seven nursery and/or finisher herds were included in this study. Oral fluids were collected by pen. Using species specific real-time PCR M. hyorhinis was detected in 97% of sampled herds, whereas 70% were positive for M. hyosynoviae. Lameness scores were determined for all pigs in each pen where oral fluids were collected. Lameness was identified in 3.9% of pigs across all sampled pens. No correlation was observed between lameness in pigs in a pen and detection of M. hyorhinis in oral fluid samples (p > 0.05), whereas a significant correlation was observed between M. hyosynoviae detection in oral fluids and lameness (p < 0.05). A negative correlation was observed between the proportion of lame pigs in the pen and Ct values for M. hyosynoviae in oral fluids (p < 0.05; r = -0.27). An age-related effect was observed with M. hyosynoviae detection in oral fluids, indicating an increased prevalence of the bacterium in finishers compared to nursery pigs. Under the conditions of this study, M. hyorhinis was frequently detected in oral fluids from nursery and finisher pigs regardless of the clinical presentation of lameness, whereas the detection of M. hyosynoviae varied depending on the age of sample pigs. Our results suggest that oral fluids may not be an informative diagnostic sample for M. hyorhinis associated lameness. However, the association of lameness and M. hyosynoviae detection in oral fluids warrants prospective population-based diagnostic studies.


Assuntos
Coxeadura Animal/complicações , Infecções por Mycoplasma/veterinária , Mycoplasma hyorhinis/genética , Mycoplasma hyosynoviae/genética , Saliva/microbiologia , Doenças dos Suínos/microbiologia , Fatores Etários , Animais , Coxeadura Animal/microbiologia , Infecções por Mycoplasma/complicações , Infecções por Mycoplasma/microbiologia , Reação em Cadeia da Polimerase em Tempo Real/veterinária , Suínos
5.
Vet Microbiol ; 237: 108408, 2019 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-31585650

RESUMO

Information on the epidemiology of C. difficile infection (CDI) in South-East Asian countries is limited, as is data on possible animal reservoirs of C. difficile in the region. We investigated the prevalence and molecular epidemiology of C. difficile in piglets and the piggery environment in Thailand and Malaysia. Piglet rectal swabs (n = 224) and piggery environmental specimens (n = 23) were collected between 2015 and 2016 from 11 farms located in Thailand and Malaysia. All specimens were tested for the presence of C. difficile with toxigenic culture. PCR assays were performed on isolates to determine the ribotype (RT), and the presence of toxin genes. Whole genome sequencing was used on a subset of isolates to determine the evolutionary relatedness of RT038 (the most prevalent RT identified) common to pigs and humans from Thailand and Indonesia. C. difficile was recovered from 35% (58/165) and 92% (54/59) of the piglets, and 89% (8/9) and 93% (13/14) of the environmental specimens from Thailand and Malaysia, respectively. All strains from Thailand, and 30 strains from Malaysia (23 piglet and 7 environmental isolates) were non-toxigenic. To our knowledge, this is the first and only report with a complete lack of toxigenic C. difficile among piglets, a feature which could have a protective effect on the host. The most common strain belonged to RT038 (ST48), accounting for 88% (51/58) of piglet and 78% (7/9) of environmental isolates from Thailand, and all 30 isolates tested from Malaysia. Piglet RT038 isolates from Thailand and Malaysia differed by only 18 core-genome single nucleotide variants (cgSNVs) and both were, on average, 30 cgSNVs different from the human strains from Thailand and Indonesia, indicating a common ancestor in the last two decades.


Assuntos
Infecções por Clostridium/veterinária , Clostridium difficile/genética , Doenças dos Suínos/microbiologia , Animais , Infecções por Clostridium/epidemiologia , Infecções por Clostridium/microbiologia , Microbiologia Ambiental , Variação Genética , Abrigo para Animais , Malásia/epidemiologia , Epidemiologia Molecular , Prevalência , Suínos , Tailândia/epidemiologia , Sequenciamento Completo do Genoma
6.
Vet Microbiol ; 237: 108419, 2019 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-31585655

RESUMO

Diarrhea is one of the major causes of neonatal mortality in pigs. In the present study, 31 pig farms with outbreaks of neonatal diarrhea were investigated in Catalonia (NE Spain) from February 2017 until June 2018. Two hundred and fifteen diarrheic samples from 1 to 7 days old piglets were tested for a panel of enteric pathogens. In 19 of the studied farms additional fecal samples from apparently healthy pen-mates were collected and tested for the same panel of infectious agents. Samples were bacteriologically cultured and tested by PCR for E. coli virulence factors genes, C. perfringens types A and C toxins (Cpα, Cpß, Cpß2) and C. difficile toxins (TcdA, TcdB). Moreover, Rotavirus A (RVA), Rotavirus B (RVB), Rotavirus C (RVC), porcine epidemic diarrhea virus (PEDV) and transmissible gastroenteritis virus (TGEV) were also determined by RT-qPCR. More than one pathogen could be detected in all of the outbreaks. Nevertheless, RVA was the only agent that could be statistically correlated with the outcome of diarrhea. For the other viruses and bacteria analyzed significant differences between the diseased pigs and the controls were not found. In spite of this, the individual analysis of each of the studied farms indicated that other agents such as RVB, RVC, toxigenic C. difficile or pathogenic E. coli could play a relevant role in the outbreak of diarrhea. In conclusion, the large diversity of agent combinations and disease situations detected in neonatal diarrhea outbreaks of this study stand for a more personalized diagnosis and management advice at a farm level.


Assuntos
Animais Recém-Nascidos , Bactérias/isolamento & purificação , Diarreia/veterinária , Enterite/veterinária , Doenças dos Suínos/microbiologia , Vírus/isolamento & purificação , Animais , Infecções Bacterianas/epidemiologia , Infecções Bacterianas/microbiologia , Infecções Bacterianas/veterinária , Diarreia/microbiologia , Surtos de Doenças/veterinária , Enterite/epidemiologia , Enterite/microbiologia , Fazendas , Fezes/microbiologia , Prevalência , Espanha/epidemiologia , Suínos , Doenças dos Suínos/epidemiologia , Fatores de Virulência , Viroses/epidemiologia , Viroses/veterinária , Viroses/virologia
7.
Vet Res ; 50(1): 85, 2019 Oct 22.
Artigo em Inglês | MEDLINE | ID: mdl-31640784

RESUMO

The causative agent of ileitis, Lawsonia intracellularis, is commonly associated with diarrhea and reduced weight gain in growing pigs. The effect of in-feed probiotics on L. intracellularis infection dynamics was evaluated. In brief, 70 2.5-week-old-pigs were randomly divided into six groups with 10-20 pigs each. All pigs were fed an age appropriate base ration for the duration of the study, which was supplemented with one of three Bacillus strains including B. amyloliquefaciens (T01), B. licheniformis (T02) and B. pumilus (T03). Another group was orally vaccinated with a commercial live L. intracellularis vaccine (VAC) at 3 weeks of age. At 7 weeks of age, T01-LAW, T02-LAW, T03-LAW, VAC-LAW and the POS-CONTROL groups were challenged with L. intracellularis while the NEG-CONTROL pigs were not challenged. All pigs were necropsied 16 days later. By the time of inoculation, all VAC-LAW pigs had seroconverted and at necropsy 10-65% of the pigs in all other challenged groups were also seropositive. The results indicate a successful L. intracellularis challenge with highest bacterial DNA levels in POS-CONTROL pigs, VAC-LAW pigs and T01-LAW pigs. There was a delay in onset of shedding in T02-LAW and T03-LAW groups, which was reflected in less severe macroscopic and microscopic lesions, reduced intralesional L. intracellularis antigen levels and a lower area under the curve for bacterial shedding. Under the study conditions, two of the probiotics tested suppressed L. intracellularis infection. The obtained findings show the potential of probiotics in achieving antibiotic-free control of L. intracellularis.


Assuntos
Bacillus pumilus/química , Derrame de Bactérias/efeitos dos fármacos , Infecções por Desulfovibrionaceae/veterinária , Lawsonia (Bactéria)/efeitos dos fármacos , Probióticos/farmacologia , Doenças dos Suínos/tratamento farmacológico , Ração Animal/análise , Animais , Bacillus amyloliquefaciens/química , Bacillus licheniformis/química , Infecções por Desulfovibrionaceae/tratamento farmacológico , Infecções por Desulfovibrionaceae/microbiologia , Infecções por Desulfovibrionaceae/patologia , Dieta/veterinária , Lawsonia (Bactéria)/fisiologia , Distribuição Aleatória , Suínos , Doenças dos Suínos/microbiologia , Doenças dos Suínos/patologia
8.
Emerg Microbes Infect ; 8(1): 1524-1527, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31631781

RESUMO

The recent emergence of plasmid-mediated tigecycline resistance genes, tet(X3) and tet(X4), in animals and humans in China would pose a foreseeable threat to public health. To illustrate this paradigm shift in tigecycline resistance, here, covering the period 2008-2018, we retrospectively analysed a national strain collection of Escherichia coli (n = 2254), obtained from chickens and pigs, in six representative provinces of China. The gene tet(X4) was identified in five pig isolates collected in 2016 and 2018 from the provinces of Sichuan (3/15, 2018), Henan (1/25, 2018) and Guangdong (1/28, 2016), but not in the isolates prior to 2016. None of the isolates was detected harbouring tet(X3). All tet(X4)-positive E. coli exhibited high levels of tigecycline resistance (MICs, 16-64 mg/L), and two were confirmed as colistin resistant, harbouring chromosome-borne mcr-1 gene. The gene tet(X4) was detected on a plasmid in all five isolates, whereas a co-location of tet(X4) on the chromosome of one isolate was observed. Diverse host strains and novel plasmids related to the tet(X4) gene were observed. Our timely findings of the recent emergence of tet(X4) gene in food animal support the rapid surveillance and eradication of this gene before it is established.


Assuntos
Antibacterianos/farmacologia , Farmacorresistência Bacteriana , Infecções por Escherichia coli/veterinária , Proteínas de Escherichia coli/genética , Escherichia coli/isolamento & purificação , Doenças das Aves Domésticas/microbiologia , Doenças dos Suínos/microbiologia , Tigeciclina/farmacologia , Animais , Galinhas , China , Colistina/farmacologia , Escherichia coli/efeitos dos fármacos , Escherichia coli/genética , Escherichia coli/metabolismo , Infecções por Escherichia coli/microbiologia , Proteínas de Escherichia coli/metabolismo , Plasmídeos/genética , Plasmídeos/metabolismo , Estudos Retrospectivos , Suínos
9.
BMC Vet Res ; 15(1): 342, 2019 Oct 16.
Artigo em Inglês | MEDLINE | ID: mdl-31619295

RESUMO

BACKGROUND: The objective of this study was to assess the efficacy of a trivalent vaccine mixture and compare it to the respective monovalent vaccines against Mycoplasma hyopneumoniae, porcine circovirus type 2 (PCV2), and porcine reproductive and respiratory syndrome virus (PRRSV). RESULTS: Pigs that were triple challenged with M. hyopneumoniae, PCV2, and PRRSV following vaccination with the trivalent vaccine mixture exhibited a significantly better growth performance when compared to unvaccinated and challenged pigs. A statistical difference was not found when comparing pig populations which were vaccinated with the trivalent vaccine followed by a triple challenge and pigs vaccinated with monovalent M hyopneumoniae vaccine followed by mycoplasmal single challenge in the following areas: M. hyopneumoniae nasal shedding, the number of M. hyopneumoniae-specific interferon-γ secreting cells (IFN-γ-SC), and mycoplasmal lung lesion scores. Pigs vaccinated with the trivalent vaccine mixture followed by a triple challenge resulted in a similar reduction of PCV2 viremia, an increase in the number of PCV2-specific IFN-γ-SC and reduction in interstitial lung lesion scores when compared to pigs vaccinated with a PCV-2 vaccine and challenged with PCV2 only. Lastly, there was a significant difference in the reduction of PRRSV viremia, an increase in PRRSV-specific IFN-γ-SC and a reduction of interstitial lung lesion scores between pigs vaccinated with the trivalent vaccine mixture followed by a triple challenge and pigs vaccinated with a monovalent PRRSV vaccine followed by PRRSV challenge only. CONCLUSION: The trivalent vaccine mixture was efficacious against a triple challenge of M. hyopneumoniae, PCV2, and PRRSV. The trivalent vaccine mixture, however, did not result in equal protection when compared against each respective monovalent vaccine, with the largest vaccine occurring within PRRSV.


Assuntos
Circovirus/imunologia , Mycoplasma hyopneumoniae/imunologia , Vírus da Síndrome Respiratória e Reprodutiva Suína/imunologia , Doenças dos Suínos/prevenção & controle , Vacinação/veterinária , Animais , Vacinas Bacterianas/imunologia , Infecções por Circoviridae/imunologia , Infecções por Circoviridae/prevenção & controle , Feminino , Masculino , Pneumonia Suína Micoplasmática/imunologia , Pneumonia Suína Micoplasmática/prevenção & controle , Síndrome Respiratória e Reprodutiva Suína/imunologia , Síndrome Respiratória e Reprodutiva Suína/prevenção & controle , Sus scrofa , Suínos , Doenças dos Suínos/imunologia , Doenças dos Suínos/microbiologia , Doenças dos Suínos/virologia , Vacinas Combinadas/administração & dosagem , Vacinas Combinadas/imunologia , Vacinas Virais/imunologia
10.
J Vet Diagn Invest ; 31(6): 818-827, 2019 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-31646949

RESUMO

Rapid identification of the infecting Salmonella serovar from porcine diagnostic samples is vital to allow implementation of appropriate on-farm treatment and management decisions. Although identification at the serogroup level can be rapidly achieved at most veterinary diagnostic laboratories, final Salmonella serovar identification often takes several weeks because of the limited number of reference laboratories performing the complex task of serotyping. Salmonella serogroup B, currently the dominant serogroup identified from swine clinical samples in the United States, contains serovars that vary from highly pathogenic to minimally pathogenic in swine. We determined the frequency of detection of individual group B serovars at the Iowa State Veterinary Diagnostic Laboratory from 2008 to 2017, and validated a multiplex real-time PCR (rtPCR) to distinguish pathogenic serogroup B serovars from those of lesser pathogenicity. Our results indicate that, since 2014, Salmonella enterica ssp. enterica serovar 4,[5],12:i:- has been the dominant serovar identified from swine clinical samples at the ISU-VDL, with S. Typhimurium now the second most common serovar identified. We developed a rtPCR to allow rapid differentiation of samples containing S. 4,[5],12:i:- and S. Typhimurium from samples containing serovars believed to be of less pathogenicity, such as S. Agona and S. Derby. When combined with enrichment culture, this rtPCR has the ability to significantly improve the time to final serovar identification of the 2 most commonly identified pathogenic Salmonella serovars in swine, and allows rapid implementation of serovar-specific intervention strategies.


Assuntos
Reação em Cadeia da Polimerase em Tempo Real/veterinária , Salmonelose Animal/diagnóstico , Salmonella enterica/classificação , Sorotipagem/veterinária , Doenças dos Suínos/diagnóstico , Animais , Reação em Cadeia da Polimerase Multiplex/métodos , Reação em Cadeia da Polimerase Multiplex/veterinária , Reação em Cadeia da Polimerase em Tempo Real/métodos , Salmonelose Animal/microbiologia , Salmonella enterica/patogenicidade , Sorogrupo , Sorotipagem/métodos , Suínos , Doenças dos Suínos/microbiologia , Estados Unidos , Virulência
11.
Infect Immun ; 87(12)2019 12.
Artigo em Inglês | MEDLINE | ID: mdl-31548316

RESUMO

Swine erysipelas is caused by the Gram-positive pathogen Erysipelothrix rhusiopathiae The swine erysipelas live vaccine in Japan, the E. rhusiopathiae Koganei 65-0.15 strain (Koganei), has been reported to cause arthritis and endocarditis. To develop a vaccine with increased safety, we used a virulent Fujisawa strain to construct transposon mutants for a total of 651 genes, which covered 38% of the coding sequence of the genome. We screened the mutants for attenuation by inoculating mice with 108 CFU of each mutant and subsequently assessed protective capability by challenging the surviving mice with 103 CFU (102 times the 50% lethal dose) of the Fujisawa strain. Of the 23 attenuated mutants obtained, 6 mutants were selected and evaluated for protective capability in pigs by comparison to that of the Koganei strain. A mutant in the ERH_0432 (tagF) gene encoding a putative CDP-glycerol glycerophosphotransferase was found to be highly attenuated and to induce humoral and cell-mediated immune responses in conventional pigs. An in-frame deletion mutant of the gene, the Δ432 mutant, was constructed, and attenuation was further confirmed in germfree piglets; three of four piglets subcutaneously inoculated with 109 CFU of the Δ432 mutant showed no apparent clinical symptoms, whereas all four of the Koganei-inoculated piglets died 3 days after inoculation. It was confirmed that conventional pigs inoculated orally or subcutaneously with the Δ432 strain were almost completely protected against lethal challenge infection. Thus, the tagF homolog mutant of E. rhusiopathiae represents a safe vaccine candidate that can be administered via the oral and subcutaneous routes.


Assuntos
Vacinas Bacterianas/imunologia , Infecções por Erysipelothrix/prevenção & controle , Erysipelothrix/genética , Erysipelothrix/imunologia , Doenças dos Suínos/prevenção & controle , Transferases (Outros Grupos de Fosfato Substituídos)/genética , Animais , Elementos de DNA Transponíveis/genética , Erysipelothrix/patogenicidade , Infecções por Erysipelothrix/imunologia , Feminino , Camundongos , Suínos , Doenças dos Suínos/imunologia , Doenças dos Suínos/microbiologia , Vacinas Atenuadas/imunologia
12.
Lett Appl Microbiol ; 69(5): 373-378, 2019 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-31506974

RESUMO

Extended spectrum ß-lactamases (ESBL) producing Shiga toxin producing E. coli (STEC) are posing a constant threat to public health throughout the world leading to serious infections and raising key therapeutic issues. A total of 219 fecal samples were collected from piglets with diarrheoa, pig farmers and water sources in North East India; and were processed for isolation of Escherichia coli. The isolates were screened for antimicrobial resistance and suspected isolates for ESBLs production by double-disk synergy test (DDST). Escherichia coli isolates positive for DDST were subjected for detection of selected ESBL/beta-lactamase genes and virulence associated genes by PCR. By DDST, 337 (67·94%) E. coli isolates were detected as ESBLs producer, of which 211 (66·98%), 117 (70·91%) and 9 (56·25%) isolates were from piglets, humans and water sources respectively. A total of 64 (12·90%) isolates were recorded as STEC, of which 48 (9·68%), 6 (1·21%) and 10 (2·02%) were from human, piglets and water respectively. Majority of the STEC isolates (64·06%) possessed multiple virulence genes, of which 59·38% also harboured ESBL/beta-lactamase genes with 32·81% STEC isolates being positive for multiple ESBL/beta-lactamase genes. SIGNIFICANCE AND IMPACT OF THE STUDY: Multidrug resistant (MDR) enteric bacteria are global concern. Association of MDR traits in STEC isolates are another rising issue in human and animal health perspective. The interaction of such organisms among the human, domestic animals and adjoining water sources require to be analysed systematically. The present study exhibited the possible transmission of MDR-STEC among the human, domestic animals and water sources in the North eastern states of India. To the best of our knowledge, this is the first report of such kind in India.


Assuntos
Infecções por Escherichia coli/microbiologia , Infecções por Escherichia coli/veterinária , Água Doce/microbiologia , Escherichia coli Shiga Toxigênica/isolamento & purificação , Doenças dos Suínos/microbiologia , beta-Lactamases/metabolismo , Animais , Antibacterianos/farmacologia , Farmacorresistência Bacteriana Múltipla , Fezes/microbiologia , Humanos , Índia , Toxina Shiga/metabolismo , Escherichia coli Shiga Toxigênica/efeitos dos fármacos , Escherichia coli Shiga Toxigênica/genética , Escherichia coli Shiga Toxigênica/metabolismo , Suínos , beta-Lactamases/genética
13.
BMC Vet Res ; 15(1): 327, 2019 Sep 12.
Artigo em Inglês | MEDLINE | ID: mdl-31511007

RESUMO

BACKGROUND: Mycoplasma hyopneumoniae (M. hyopneumoniae) is the primary pathogen of porcine enzootic pneumonia, which has been associated with economic losses due to reduced daily weight gain and feed efficiency. Although it has a small genome and no more than 1000 genes, M. hyopneumoniae can be cultured in cell free media. However, some proteins were not expressed or were only expressed in negligible amounts under culture conditions. Nevertheless, some of these proteins can be expressed at a high level and induce a strong and rapid immune response after M. hyopneumoniae infection. The unexpressed or less expressed proteins may play critical roles in pathogenesis and/or immune response. In order to find the differentially expressed proteins of M. hyopneumoniae between culture condition and infected animals, we established an indirect ELISA for the detection of humoral immunodominant proteins which can discriminate between inactivated bacterin-induced hyperimmune sera and convalescent sera by using Mhp366 protein which did not react with sera from bacterin-immunized pigs, but revealed a strong immunoreaction with porcine convalescent sera. RESULTS: The checkerboard titration method was done by using porcine convalescent sera as positive sera and inactivated bacterin-induced hyperimmune sera as negative sera. The bacterial lysates of fusion proteins and free GST protein without dilution were the optimal coating antigens. The optimal blocking buffer was PBS with 10% FBS and 2.5% skimmed milk. In the checkboard ELISAs, when the sera were diluted at 1:500 and the HRP-labeled rabbit anti-pig IgG were diluted at 1:20000, most positive result was obtained for the assay. CONCLUSIONS: This established indirect ELISA can be used as a tool for the detection of humoral immunodominant proteins of M. hyopneumoniae which can discriminate between inactivated bacterin-induced hyperimmune sera and convalescent sera.


Assuntos
Vacinas Bacterianas/imunologia , Ensaio de Imunoadsorção Enzimática/veterinária , Mycoplasma hyopneumoniae/imunologia , Pneumonia Suína Micoplasmática/imunologia , Animais , Proteínas de Bactérias/imunologia , Vacinas Bacterianas/administração & dosagem , Vacinas Bacterianas/sangue , Ensaio de Imunoadsorção Enzimática/métodos , Mycoplasma hyopneumoniae/química , Pneumonia Suína Micoplasmática/sangue , Suínos , Doenças dos Suínos/microbiologia
14.
J Anim Sci ; 97(11): 4503-4508, 2019 Nov 04.
Artigo em Inglês | MEDLINE | ID: mdl-31545364

RESUMO

An infection model with enterotoxigenic Escherichia coli (ETEC) harboring the F4 fimbriae can be used to assess the impacts that various challenges associated with weaning (e.g., dietary, psychological, environmental) have on the expression of postweaning diarrhea. The objective of this study was to develop a novel inoculation method for administering an ETEC culture that would induce a higher proportion of ETEC-F4 diarrhea, in pigs that genetically showed ETEC-F4 susceptibility or resistance. The study was designed as a factorial arrangement of treatments with the factors being 1) partially susceptible or resistant to ETEC-F4 based on genetic testing, and 2) 4 challenge treatments, being a) a conventional liquid broth method using a drenching gun [Positive control (PC)], b) a Syringe method, c) a Capsule method, and d) Negative control [pigs not challenged (NC)]. At 21 ± 3 d of age (mean ± SEM), 48 male castrate pigs (Large White × Landrace) weighing approximately 7.0 ± 1.18 kg were allocated to 4 treatment groups in 2 replicate pens (6 pigs per pen). Initial ETEC-F4 susceptibility was based on a DNA marker test and each treatment group had 9 partially susceptible and 3 resistant pigs. On days 7 and 8 after weaning, pigs were challenged with ETEC (serotype O149:K88; toxins LT1, ST1, ST2, and EAST). On each inoculation day the PC pigs were orally dosed with 9 mL 7.12 × 109 colony-forming unit (CFU), the Syringe pigs with 0.8 mL 6.72 × 109 CFU, the Capsule pigs were orally administered 2 capsules containing 0.8 mL 3.28 × 109 CFU, and the NC pigs 1 mL of phosphate-buffered saline (PBS) solution. Approximately 72 h after infection, 44, 22, 78, and 0% of partially susceptible pigs in the PC, the Syringe, the Capsule, and the NC group had developed ETEC-F4 diarrhea (P = 0.007). Partially susceptible pigs had a higher diarrhea index (DI) compared to resistant pigs (31.5 vs. 4.8, P < 0.001). The NC group had a lower DI compared to the PC and Capsule pigs (3.9, 38.1, and 40.3, respectively, P < 0.005). Following infection, genetically resistant pigs in the Capsule group had a DI of zero and the partially susceptible pigs had a DI of 55.6 (P = 0.014). This study showed that genetically screening pigs and using a Capsule to deliver ETEC-F4 can increase cases of diarrhea and the efficiency of the challenge model. Taken together, these methods have the potential to reduce the number of pigs needed in future experimental infection studies.


Assuntos
Diarreia/veterinária , Modelos Animais de Doenças , Escherichia coli Enterotoxigênica/fisiologia , Infecções por Escherichia coli/veterinária , Doenças dos Suínos/microbiologia , Suínos , Animais , Infecções por Escherichia coli/microbiologia , Loci Gênicos , Masculino , Desmame
15.
Int J Food Microbiol ; 310: 108310, 2019 Nov 16.
Artigo em Inglês | MEDLINE | ID: mdl-31476581

RESUMO

Pigs infected with Salmonella are an important source of contamination at slaughterhouses. We characterized the distribution, virulence genotypes and antimicrobial-resistance phenotypes for Salmonella isolates that were collected from different stages of a pork production chain. Each of ten pig lots were sampled for feed (n = 10), water (n = 10), barn floor (n = 10), lairage floor (n = 10), mesenteric lymph nodes (n = 100), tonsils (n = 100), processing environment (n = 120), pork cuts (n = 40) and carcasses after bleeding (n = 100), after singeing (n = 100), after evisceration (n = 100), and after final rinsing (n = 100). Salmonella was isolated according to ISO 6579, and after confirmation the isolates were subjected to serogrouping, macro-restriction digests and pulsed-field gel electrophoresis (PFGE), detection of virulence-related genes and antimicrobial-resistance phenotyping. Salmonella was recovered from barn floors from 3 pig farms (3/10), lairage floors (7/10), carcasses after bleeding (2/100) and final washing (1/100), palatine tonsils (45/100), mesenteric lymph nodes (43/100), utensils (3/120) and cuts (4/40). The most prevalent serogroup was O: 4 (82%) followed by O:3 (7.7%); O:9 (5.1%); O:8 (2.6%) and O:7 (2.6%). Recovered strains (n = 109) were classified into 24 different pulsotypes (XbaI restriction digest), which were arranged into five different clusters. Fourteen different virulence genotypes were observed based on 15 loci, and all isolates were positive for invA, sitC, pagC and tolC. There was a high prevalence of antimicrobial resistance against streptomycin (90.5%), tetracycline (88.1%), ampicillin (81.0%), chloramphenicol (71.4%), and ciprofloxacin (50.0%). No strain was resistant to ertapenem, meropenem or kanamycin. A majority (80.9%) of isolates were considered multidrug resistant (resistant to ≥3 antibiotic classes). This study provides valuable insight about the epidemiology of Salmonella in swine production, and despite the low presence of this pathogen in carcasses and meat cuts, the majority of isolates was multidrug resistant.


Assuntos
Resistência Microbiana a Medicamentos/genética , Variação Genética , Salmonella , Virulência/genética , Matadouros , Animais , Antibacterianos/farmacologia , Brasil , Eletroforese em Gel de Campo Pulsado , Genótipo , Carne/microbiologia , Tonsila Palatina/microbiologia , Prevalência , Salmonella/classificação , Salmonella/efeitos dos fármacos , Salmonella/genética , Salmonella/isolamento & purificação , Salmonelose Animal/microbiologia , Sorogrupo , Suínos , Doenças dos Suínos/microbiologia
16.
Vet Res ; 50(1): 69, 2019 Sep 23.
Artigo em Inglês | MEDLINE | ID: mdl-31547880

RESUMO

Haemophilus parasuis is part of the microbiota of the upper respiratory tract in swine. However, virulent strains can cause a systemic disease known as Glässer's disease. Several virulence factors have been described in H. parasuis including the virulence-associated trimeric autotransporters (VtaAs). VtaA2 is up-regulated during infection and is only found in virulent strains. In order to determine its biological function, the vtaA2 gene was cloned with its native promotor region in pACYC184, and the transformed Escherichia coli was used to perform functional in vitro assays. VtaA2 was found to have a role in attachment to plastic, mucin, BSA, fibronectin and collagen. As other VtaAs from H. parasuis, the passenger domain of VtaA2 contains collagen domains. In order to examine the contribution of the collagen repeats to VtaA2 function, a recombinant vtaA2 without the central collagen domains was obtained and named vtaA2OL. VtaA2OL showed similar capacity than VtaA2 to adhere to plastic, mucin, BSA, fibronectin and plasma but a reduced capacity to adhere to collagen, suggesting that the collagen domains of VtaA2 are involved in collagen attachment. No function in cell adhesion and invasion to epithelial alveolar cell line A549 or unspecific binding to primary alveolar macrophages was found. Likewise VtaA2 had no role in serum or phagocytosis resistance. We propose that VtaA2 mediates adherence to the host by binding to the mucin, found in the upper respiratory tract mucus, and to the extracellular matrix proteins, present in the connective tissue of systemic sites, such as the serosa.


Assuntos
Aderência Bacteriana/genética , Infecções por Haemophilus/veterinária , Haemophilus parasuis/fisiologia , Doenças dos Suínos/microbiologia , Fatores de Virulência/genética , Animais , Escherichia coli/genética , Infecções por Haemophilus/microbiologia , Suínos , Virulência/genética
17.
BMC Vet Res ; 15(1): 272, 2019 Aug 01.
Artigo em Inglês | MEDLINE | ID: mdl-31370843

RESUMO

BACKGROUND: Ceftiofur Sodium is widely used in China. Our aim was to determine Ceftiofur Sodium activity and optimize dosing regimens against the pathogen Haemophilus parasuis using an in vitro and ex vivo pharmacokinetics/pharmacodynamics modeling approach. By adopting these strategies, we wanted to extend the effective life of Ceftiofur Sodium in reduce drug-resistance in pigs. RESULTS: We established an H. parasuis infection model in pigs, and assessed the pharmacokinetics of Ceftiofur Sodium in both healthy and infected animals. After Ceftiofur Sodium (10 mg/kg, i.m.) administration to healthy and H. parasuis-infected pigs, plasma based desfuroylceftiofur (a metabolite of Ceftiofur Sodium) was measured by High Performance Liquid Chromatography. The pharmacokinetics of Ceftiofur Sodium (desfuroylceftiofur) was consistent with a two-compartment open model, with first-order absorption. We observed no significant differences (P > 0.05) in pharmacokinetic parameters between healthy and infected pigs. Pharmacodynamics data showed that Ceftiofur Sodium was highly inhibitory against H. parasuis, with MIC, MBC, and MPC values of 0.003125, 0.0125 and 0.032 µg/mL, respectively. Desfuroylceftiofur in plasma also had strong bactericidal activity. Almost all H. parasuis cultured in plasma medium of Ceftiofur Sodium-inoculated healthy pigs, at each time point, were killed within 24 h. A weaker antibacterial activity was measured in infected-pig plasma medium at 18, 24, 36, and 48 h, after Ceftiofur Sodium inoculation. Pharmacokinetic parameters were combined with ex vivo pharmacodynamic parameters, and the bacteriostatic effect (36.006 h), bactericidal effect (71.637 h) and clearance (90.619 h) within 24 h, were determined using the Hill equation. Dose-calculation equations revealed the optimal dose of Ceftiofur Sodium to be 0.599-1.507 mg/kg. CONCLUSIONS: There were no significant differences in Ceftiofur Sodium pharmacokinetic parameters between healthy and infected pigs, although pharmacokinetics/pharmacodynamics fitting curves showed obviously differences. The optimal dose of Ceftiofur Sodium was lower than recommended (3 mg/kg), which may provide improved treatments for Glässers disease, with lower drug-resistance possibility.


Assuntos
Cefalosporinas , Infecções por Haemophilus/veterinária , Modelos Biológicos , Animais , Antibacterianos/farmacocinética , Antibacterianos/farmacologia , Antibacterianos/uso terapêutico , Cefalosporinas/farmacocinética , Cefalosporinas/farmacologia , Cefalosporinas/uso terapêutico , Infecções por Haemophilus/tratamento farmacológico , Infecções por Haemophilus/microbiologia , Haemophilus parasuis/efeitos dos fármacos , Suínos , Doenças dos Suínos/tratamento farmacológico , Doenças dos Suínos/microbiologia
18.
Vet Microbiol ; 235: 270-279, 2019 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-31383312

RESUMO

Lawsonia intracellularis is an obligate intracellular microorganism and the causative agent of porcine proliferative enteropathy. Due to its obligate intracellular nature, characterization of antigens and proteins involved in host-pathogen interaction and immune recognition have been difficult to achieve using conventional microbiological techniques. In this work, we used 2-dimensional gel electrophoresis coupled with Western-immunoblotting, mass spectrometry and bioinformatics to identify bacterial proteins that interact in vitro with pig intestinal cells (IPEC-1), have immunogenic properties and the potential to be used as subunit vaccine antigens. We detected eleven immunogenic bacterial proteins from which fliC (LI0710), LI1153 (annotated by NCBI as Putative protein N), and LI0649 (annotated as autotransporter) were predicted to be expressed on the outer membrane while LI0169 (oppA; annotated as ABC dipeptide transport system) was predicted to be periplasmic with a transmembrane domain forming a central pore through the plasma membrane. Genes coding for these four proteins were cloned and expressed in Escherichia coli and the corresponding recombinant proteins were purified using affinity chromatography. Porcine hyperimmune serum against whole Lawsonia lysate established that all four recombinant proteins were immunogenic. Further, rabbit hyperimmune sera generated against the vaccine strain of L. intracellularis and rabbit serum specific for each recombinant protein showed an inhibitory effect on the attachment and penetration of live, avirulent L. intracellularis, thus indicating that each protein is a potential neutralizing antibody target and a candidate for subunit vaccine formulation.


Assuntos
Anticorpos Antibacterianos/imunologia , Anticorpos Neutralizantes/imunologia , Proteínas de Bactérias/imunologia , Vacinas Bacterianas/imunologia , Infecções por Desulfovibrionaceae/veterinária , Lawsonia (Bactéria)/imunologia , Animais , Proteínas de Bactérias/genética , Western Blotting , Linhagem Celular , Biologia Computacional , Infecções por Desulfovibrionaceae/imunologia , Infecções por Desulfovibrionaceae/prevenção & controle , Feminino , Intestinos/citologia , Intestinos/microbiologia , Espectrometria de Massas , Proteômica , Coelhos , Proteínas Recombinantes/imunologia , Suínos , Doenças dos Suínos/imunologia , Doenças dos Suínos/microbiologia , Doenças dos Suínos/prevenção & controle , Vacinas de Subunidades/imunologia
20.
BMC Vet Res ; 15(1): 294, 2019 Aug 14.
Artigo em Inglês | MEDLINE | ID: mdl-31412870

RESUMO

BACKGROUND: Glaesserella parasuis (G. parasuis) is an influential pathogen of the pig, which induces high morbidity and mortality in naive pig populations in the pig industry. Accurate and rapid detection of the agent is important for disease control. In this study, a simple recombinase polymerase amplification (RPA) with a Lateral flow (LF) strip (RPA-LF-GPS) was developed to detect G. parasuis. RESULTS: The RPA-LF-GPS can specifically detect G. parasuis a limit of 100 CFU from other common related pathogens causing arthritis in the pig. The RPA-LF-GPS assay can use boiled synovial fluid samples as a template with the same sensitivity as other DNA extraction methods. In the detection of clinic positive synovial fluid sample, RPA-LF-GPS is equally sensitive (98.1%) compared with that of PCR (90.4%) (P > 0.05). The whole procedure of the RPA-LF-GPS assay could be finished in 1 hour without professional equipment. CONCLUSIONS: RPA-LF-GPS assay is a rapid and simple method for point-of-care diagnostic testing for G. parasuis infection.


Assuntos
Infecções por Haemophilus/veterinária , Haemophilus parasuis , Recombinases/metabolismo , Doenças dos Suínos/microbiologia , Líquido Sinovial/microbiologia , Animais , Infecções por Haemophilus/microbiologia , Técnicas de Amplificação de Ácido Nucleico/veterinária , Reação em Cadeia da Polimerase/veterinária , Recombinases/química , Recombinases/genética , Sensibilidade e Especificidade , Suínos , Doenças dos Suínos/diagnóstico
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