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1.
Lett Appl Microbiol ; 71(4): 394-399, 2020 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-32557652

RESUMO

The aims of the present study were to examine the occurrence of Staphylococcus spp. in the tonsils of slaughtered pigs in a regional slaughterhouse in Greece, the antibiotic resistance of the Staphylococcus spp. isolates, and the enteroxigenicity of the S. aureus isolates. Staphylococcus spp. were isolated in 70 (48·61%) out of the total 144 tonsil samples. The predominant species was S. aureus in coagulase-positive staphylococci (CoPS), while the predominant species were Staphylococcus epidermidis and Staphylococcus saprophyticus in the coagulase-negative staphylococci (CoNS). Staphylococcus spp. isolates presented high antibiotic resistance frequencies to tetracycline (97·1%) or clindamycin (80·0%) and low antibiotic resistance frequencies to fusidic acid (14·3%). No methicillin-resistant S. aureus (MRSA) strains were identified, and all Staphylococcus spp. isolates were susceptible to vancomycin. Among the 26 S. aureus isolates, 21 (80·76%) possessed staphylococcal enterotoxin genes with seven different enterotoxin gene profiles. The predominant enterotoxin profile was seg, sei and sej with seven S. aureus isolates. The occurrence of multidrug resistant Staphylococcus spp. in pig tonsils indicate public health risk to pork consumers and handlers in developing antimicrobial resistance.


Assuntos
Farmacorresistência Bacteriana , Enterotoxinas/metabolismo , Tonsila Palatina/microbiologia , Infecções Estafilocócicas/veterinária , Staphylococcus/isolamento & purificação , Doenças dos Suínos/microbiologia , Matadouros/estatística & dados numéricos , Animais , Antibacterianos/farmacologia , Clindamicina/farmacologia , Grécia/epidemiologia , Staphylococcus aureus Resistente à Meticilina/efeitos dos fármacos , Testes de Sensibilidade Microbiana , Infecções Estafilocócicas/epidemiologia , Infecções Estafilocócicas/microbiologia , Staphylococcus/classificação , Staphylococcus/efeitos dos fármacos , Staphylococcus/genética , Suínos , Doenças dos Suínos/epidemiologia
2.
PLoS One ; 15(4): e0228122, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32243441

RESUMO

The present study aimed to specify diagnostics for peritonsillar abscesses (PTAs) and to clarify the role of minor salivary glands. This prospective cohort study included 112 patients with acute tonsillitis (AT) and PTA recruited at a tertiary hospital emergency department between February and October 2017. All patients completed a questionnaire concerning their current disease. Serum amylase (S-Amyl) and C-reactive protein (S-CRP) levels, tonsillar findings, and pus aspirate samples and throat cultures were analyzed. Eight of 58 PTA patients (13.8%) had no signs of tonsillar infection. The absence of tonsillar erythema and exudate was associated with low S-CRP (p<0.001) and older age (p<0.001). We also observed an inverse correlation between S-Amyl and S-CRP levels (AT, r = -0.519; PTA, r = -0.353). Therefore, we observed a group of PTA patients without signs of tonsillar infection who had significantly lower S-CRP levels than other PTA patients. These findings support that PTA may be caused by an etiology other than AT. Variations in the S-Amyl levels and a negative correlation between S-Amyl and S-CRP levels may indicate that minor salivary glands are involved in PTA development.


Assuntos
Abscesso Peritonsilar/epidemiologia , Glândulas Salivares Menores/microbiologia , Infecções Estreptocócicas/epidemiologia , Streptococcus pyogenes/isolamento & purificação , Tonsilite/complicações , Adolescente , Adulto , Idoso , Idoso de 80 Anos ou mais , Amilases/sangue , Proteína C-Reativa/análise , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Tonsila Palatina/microbiologia , Abscesso Peritonsilar/diagnóstico , Abscesso Peritonsilar/microbiologia , Abscesso Peritonsilar/patologia , Prevalência , Estudos Prospectivos , Glândulas Salivares Menores/patologia , Infecções Estreptocócicas/diagnóstico , Infecções Estreptocócicas/microbiologia , Infecções Estreptocócicas/patologia , Tonsilite/sangue , Tonsilite/epidemiologia , Tonsilite/microbiologia , Adulto Jovem
3.
Vet Microbiol ; 240: 108532, 2020 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-31902502

RESUMO

Most outbreaks of disease due to infection with Actinobacillus (A.) pleuropneumoniae are caused by pigs already pre-colonised in tonsillar tissue, where the pathogen is protected from exposure to antibiotic substances administered for treatment. As it has been shown recently under experimental conditions, A. pleuropneumoniae displays host tissue-specific metabolic adaptation. In this study, pairs of A. pleuropneumoniae field isolates were recovered from lung as well as from tonsillar and nasal tissue from 20 pigs suffering from acute clinical signs of pleuropneumonia and showing characteristic pathological lung alterations. Metabolic adaptation to the porcine lower and upper respiratory tract of 32 A. pleuropneumoniae serotype 2 field isolates was examined using Fourier transform infrared (FTIR) spectroscopy as a high resolution metabolic fingerprinting method. All strains showed metabolic adaptations to organ tissue reflected by hierarchical cluster analysis of FTIR spectra similar to those previously observed under experimental conditions. Notably, differences in antimicrobial resistance patterns and minimal inhibitory concentrations of isolates from different tissues in the same animal, but not in biofilm production capability in a microtiter plate assay were found. Overall, biofilm formation was observed for 71 % of the isolates, confirming that A. pleuropneumoniae field isolates are generally able to form biofilms, although rather in a serotype-specific than in an organ-specific manner. A. pleuropneumoniae serotype 6 isolates formed significantly more biofilm than the other serotypes. Furthermore, biofilm production was negatively correlated to the lung lesion scores and tonsillar isolates tended to be more susceptible to antimicrobial substances with high bioavailability than lung isolates.


Assuntos
Infecções por Actinobacillus/veterinária , Actinobacillus pleuropneumoniae/metabolismo , Adaptação Fisiológica , Biofilmes/crescimento & desenvolvimento , Pleuropneumonia/veterinária , Pneumonia Bacteriana/veterinária , Doenças Respiratórias/veterinária , Infecções por Actinobacillus/microbiologia , Actinobacillus pleuropneumoniae/efeitos dos fármacos , Actinobacillus pleuropneumoniae/crescimento & desenvolvimento , Animais , Antibacterianos/farmacologia , Brônquios/microbiologia , Pulmão/microbiologia , Pulmão/patologia , Testes de Sensibilidade Microbiana , Tonsila Palatina/microbiologia , Pleuropneumonia/microbiologia , Pneumonia Bacteriana/microbiologia , Doenças Respiratórias/microbiologia , Suínos/microbiologia , Doenças dos Suínos/microbiologia
4.
Transbound Emerg Dis ; 67(1): 308-317, 2020 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-31512795

RESUMO

The number of natural infections with Mycobacterium caprae in wildlife and in cattle in the Bavarian and Austrian alpine regions has increased over the last decade. Red deer (Cervus elaphus) have been recognized as maintenance reservoir; however, the transmission routes of M. caprae among and from naturally infected red deer are unknown. The unexpected high prevalence in some hot spot regions might suggest an effective indirect transmission of infection. Therefore, this study was undertaken to diagnose the occurrence of M. caprae in faeces and secretions of red deer in their natural habitat. A total of 2,806 red deer hunted in this region during 2014-2016 were included in this study. After pathological examination, organs (lymph nodes, lung, heart), excretions and secretions (faeces, urine, saliva and tonsil swabs) were further investigated by qPCR specific for Mycobacterium tuberculosis complex (MTC), M. bovis and M. caprae. Samples tested positive by qPCR were processed for culturing of mycobacteria. In total, 55 (2.0%) animals were confirmed positive for M. caprae by pathological examination, PCR and culturing of the affected organ material. With the exception of one sample, all of the secretion and excretion samples were negative for mycobacteria of the Mycobacterium tuberculosis complex (MTC). From one red deer, M. caprae could be isolated from the heart sac as well as from the faeces. Whole-genome sequencing confirmed that both strains were clonally related. This is the first confirmation that M. caprae can be shed with the faeces of a naturally infected red deer. However, further studies focusing on a higher number of infected animals, sample standardization and coordinated multiple sampling are necessary to improve the understanding of transmission routes under natural conditions.


Assuntos
Doenças dos Bovinos/microbiologia , Cervos/microbiologia , Reservatórios de Doenças/microbiologia , Mycobacterium bovis/fisiologia , Tuberculose Bovina/microbiologia , Animais , Derrame de Bactérias , Bovinos , Doenças dos Bovinos/epidemiologia , Estudos Transversais , Fezes/microbiologia , Geografia , Alemanha/epidemiologia , Linfonodos/microbiologia , Mycobacterium bovis/classificação , Mycobacterium bovis/genética , Tonsila Palatina/microbiologia , Polimorfismo de Nucleotídeo Único , Reação em Cadeia da Polimerase em Tempo Real/veterinária , Saliva/microbiologia , Tuberculose Bovina/epidemiologia
5.
J Vet Med Sci ; 82(1): 9-13, 2020 Jan 10.
Artigo em Inglês | MEDLINE | ID: mdl-31748445

RESUMO

Streptococcus suis strains isolated from porcine endocarditis and tonsils in the Tokai area of Japan during 2004-2007 and 2014-2016 (n=114) were tested for antimicrobial susceptibility and distribution of selected resistance genes. No strains showed resistance to penicillin, ampicillin, cefotaxime, meropenem, vancomycin, and levofloxacin. High resistance to tetracycline (80.7%), clindamycin (65.8%), erythromycin (56.1%), and clarithromycin (56.1%) was observed. In chloramphenicol and sulfamethoxazole-trimethoprim, there was a trend towards increased resistance between the first (2004-2007) and second (2014-2016) periods. tet(O) and erm(B) genes were the most frequently detected, and tet(M) and mef(A/E) genes were only detected in strains isolated during 2014-2016. These results indicate that chloramphenicol and sulfamethoxazole-trimethoprim resistance, and tet(M) and mef(A/E) genes emerged in S. suis of this area after 2014.


Assuntos
Farmacorresistência Bacteriana , Infecções Estreptocócicas/veterinária , Streptococcus suis/genética , Animais , Antibacterianos/farmacologia , Endocardite Bacteriana/microbiologia , Endocardite Bacteriana/veterinária , Genótipo , Japão , Testes de Sensibilidade Microbiana , Tonsila Palatina/microbiologia , Fenótipo , Streptococcus suis/isolamento & purificação , Suínos , Doenças dos Suínos/microbiologia
6.
APMIS ; 128(3): 211-219, 2020 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-31692060

RESUMO

The aim of this study was to analyze the prevalence of Staphylococcus aureus in the tonsils of children subjected tonsillectomy due to recurrent tonsilitis and to determine the spa types of the pathogens, carriage of virulence genes and antimicrobial resistance profiles. The study included 73 tonsillectomized children. Bacteria, including S. aureus were isolated from tonsillar surface prior to tonsillectomy, recovered from tonsillar core at the time of the surgery, and from posterior pharynx 2-4 weeks after the procedure. Staphylococcus aureus isolates were compared by spa typing, tested for antimicrobial susceptibility and for the presence of superantigenic toxin genes (sea-seu, eta, etb, tst, lukS/lukF-PV) by multiplex polymerase chain reaction. Seventy-three patients (mean 7.1 ± 4.1 years, 61.6% male) were assessed. The most commonly isolated bacteria were S. aureus. The largest proportion of staphylococcal isolates originated from tonsillar core (63%), followed by tonsillar surface (45.1%) and posterior pharynx in tonsillectomized children (18.2%, p = 0.007). Five (6.3%) isolates were identified as MRSA (mecA-positive). Up to 67.5% of the isolates synthesized penicillinases (blaZ-positive isolates), and 8.8% displayed MLSB resistance. The superantigenic toxin genes were detected in more than half of examined isolates (56.3%). spa types t091, t084, and t002, and clonal complexes (CCs) CC7, CC45, and CC30 turned out to be most common. Staphylococcus aureus associated with RT in children showed pathogenicity potential and considerable genetic diversity, and no clones were found to be specific for this condition although further studies are needed.


Assuntos
Farmacorresistência Bacteriana/genética , Variação Genética/genética , Staphylococcus aureus/genética , Tonsilite/microbiologia , Adolescente , Anti-Infecciosos/uso terapêutico , Criança , Pré-Escolar , Feminino , Humanos , Masculino , Testes de Sensibilidade Microbiana/métodos , Tonsila Palatina/microbiologia , Prevalência , Infecções Estafilocócicas/tratamento farmacológico , Infecções Estafilocócicas/microbiologia , Staphylococcus aureus/efeitos dos fármacos , Tonsilectomia/métodos , Tonsilite/tratamento farmacológico , Virulência/genética , Fatores de Virulência/genética
7.
Genes (Basel) ; 10(12)2019 11 28.
Artigo em Inglês | MEDLINE | ID: mdl-31795199

RESUMO

Salmonella is one of the most common food-borne pathogens. It can be transmitted between chickens, as well as to people by contaminated poultry products. In our study, we distinguished chickens with different resistances mainly based on bacterial loads. We compared the cecal tonsil transcriptomes between the susceptible and resistant chickens after Salmonella infection, aiming to identify the crucial genes participating in the antibacterial activity in the cecal tonsil. A total of 3214 differentially expressed genes (DEGs), including 2092 upregulated and 1122 downregulated genes, were identified between the two groups (fold change ≥ 2.0, padj < 0.05). Many DEGs were mainly involved in the regulation of two biological processes: crosstalk between the cecal tonsil epithelium and pathogenic bacteria, such as focal adhesion, extracellular-matrix-receptor interaction, and regulation of the actin cytoskeleton and host immune response including the cytokine-receptor interaction. In particular, the challenged resistant birds exhibited strong activation of the intestinal immune network for IgA production, which perhaps contributed to the resistance to Salmonella infection. These findings give insight into the mRNA profile of the cecal tonsil between the two groups after initial Salmonella stimulation, which may extend the known complexity of molecular mechanisms in chicken immune response to Salmonella.


Assuntos
Resistência à Doença , Perfilação da Expressão Gênica/veterinária , Tonsila Palatina/microbiologia , Salmonella typhimurium/imunologia , Animais , Estudos de Casos e Controles , Galinhas , Regulação da Expressão Gênica , Redes Reguladoras de Genes , Tonsila Palatina/química , Tonsila Palatina/imunologia , Análise de Sequência de RNA/veterinária
8.
Sci Rep ; 9(1): 20130, 2019 12 27.
Artigo em Inglês | MEDLINE | ID: mdl-31882880

RESUMO

Streptococcus mutans is known to be a major causative agent of dental caries, and strains expressing the cell surface collagen-binding Cnm protein contribute to the development of several systemic diseases. A relationship between tonsillar immunity and glomerulonephritis has been recognized in IgA nephropathy (IgAN), and specific pathogens may have effects on tonsillar immunity (mucosal immunity). Here, we present findings showing a relationship between the presence of Cnm-positive S. mutans strains in the tonsils of IgAN patients and IgAN condition/pathogenesis. Analyses of tonsillar specimens obtained from patients with IgAN (n = 61) and chronic tonsillitis (controls; n = 40) showed that the Cnm protein-positive rate was significantly higher in IgAN patients. Among IgAN patients, the tonsillar Cnm-positive group (n = 15) had a significantly higher proportion of patients with high urinary protein (>1.5 g/gCr) and lower serum albumin level than the Cnm-negative group (n = 46). Additionally, Cnm protein and CD68, a common human macrophage marker, were shown to be merged in the tonsils of IgAN patients. These findings suggest that Cnm-positive S. mutans strains in the tonsils may be associated with severe IgAN.


Assuntos
Suscetibilidade a Doenças , Glomerulonefrite por IGA/etiologia , Tonsila Palatina/imunologia , Tonsila Palatina/microbiologia , Streptococcus mutans/imunologia , Adulto , Biomarcadores , Biópsia , Suscetibilidade a Doenças/imunologia , Feminino , Glomerulonefrite por IGA/diagnóstico , Glomerulonefrite por IGA/metabolismo , Humanos , Masculino , Pessoa de Meia-Idade , Tonsila Palatina/patologia , Infecções Estreptocócicas/complicações , Infecções Estreptocócicas/imunologia , Infecções Estreptocócicas/microbiologia , Tonsilite/complicações , Tonsilite/imunologia , Tonsilite/microbiologia , Tonsilite/patologia
9.
J Clin Microbiol ; 58(1)2019 12 23.
Artigo em Inglês | MEDLINE | ID: mdl-31694973

RESUMO

The aim of this study was to determine whether Chlamydia trachomatis could be detected in saliva and if infection is specific to an anatomical site in the oropharynx. Men who have sex with men (MSM) who were diagnosed with oropharyngeal chlamydia at the Melbourne Sexual Health Centre in 2017-2018 were invited to participate upon returning for treatment. Swabs at the tonsillar fossae and posterior oropharynx and a saliva sample were collected. Throat samples were tested for C. trachomatis by the Aptima Combo 2 assay. The bacterial loads of C. trachomatis in all samples were assessed by quantitative PCR (qPCR) detecting the ompA gene. We calculated the positivity and bacterial load of C. trachomatis for all samples. Forty-two MSM were included. The median age was 28 years (interquartile range [IQR], 24 to 33 years). Thirty-two participants (76.2%; 95% confidence interval [CI], 60.5% to 87.9%) had C. trachomatis detected by qPCR at both the tonsillar fossae and the posterior oropharynx, followed by 9.5% (n = 4; 95% CI, 2.7% to 22.6%) positive at the posterior oropharynx only and 4.8% (n = 2; 95% CI, 0.58% to 16.2%) positive at the tonsillar fossae only. Twenty-nine MSM had C. trachomatis detected in saliva (69.0%; 95% CI, 52.9% to 82.3%). The median C. trachomatis load in saliva was 446 copies/ml (IQR, 204 to 1,390 copies/ml), that in the tonsillar fossae was 893 copies/swab (IQR, 390 to 13,224 copies/ml), and that in the posterior oropharynx was 1,204 copies/swab (IQR, 330 to 16,211). There was no significant difference in C. trachomatis load between the tonsillar fossae and the posterior oropharynx (P = 0.119). Among MSM with oropharyngeal chlamydia, nearly three-quarters had chlamydia DNA detected in saliva, although the viability and implications for transmission are unknown.


Assuntos
Carga Bacteriana , Portador Sadio/epidemiologia , Portador Sadio/microbiologia , Infecções por Chlamydia/microbiologia , Chlamydia trachomatis , Homossexualidade Masculina , Faringite/epidemiologia , Faringite/microbiologia , Austrália/epidemiologia , Infecções por Chlamydia/epidemiologia , Chlamydia trachomatis/classificação , Chlamydia trachomatis/isolamento & purificação , Humanos , Masculino , Tonsila Palatina/microbiologia , Vigilância da População , Saliva/microbiologia , Comportamento Sexual
10.
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
11.
Eur J Clin Microbiol Infect Dis ; 38(10): 1901-1906, 2019 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-31292789

RESUMO

Erysipelas is a common skin infection causing significant morbidity. At present there are no established procedures for bacteriological sampling. Here we investigate the possibility of using cultures for diagnostic purposes by determining the perianal colonization with beta-hemolytic streptococci (BHS) in patients with erysipelas. Patients with erysipelas and a control group of patients with fever without signs of skin infection were prospectively included and cultures for BHS were taken from the tonsils, the perianal area, and wounds. BHS were grouped according to Lancefield antigen, species-determined, and emm-typed. Renewed cultures were taken after four weeks from patients with erysipelas and a positive culture for BHS. 25 patients with erysipelas and 25 with fever were included. In the group with erysipelas, 11 patients (44%) were colonized with BHS, ten patients were colonized in the perianal area, and one patient in the throat. In contrast, only one patient in the control group was colonized (p = 0.005 for difference). All of the patients with erysipelas colonized with BHS had an erythema located to the lower limb. The BHS were then subjected to MALDI-TOF MS and most commonly found to be Streptococcus dysgalactiae. Renewed cultures were taken from nine of the 11 patients with BHS and three of these were still colonized. Streptococcus dysgalactiae colonizes the perianal area in a substantial proportion of patients with erysipelas. The possibility of using cultures from this area as a diagnostic method in patients with erysipelas seems promising.


Assuntos
Portador Sadio/microbiologia , Erisipela/microbiologia , Streptococcus/isolamento & purificação , Adolescente , Adulto , Idoso , Idoso de 80 Anos ou mais , Canal Anal/microbiologia , Portador Sadio/epidemiologia , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Tonsila Palatina/microbiologia , Períneo/microbiologia , Prevalência , Estudos Prospectivos , Ferimentos e Lesões/microbiologia , Adulto Jovem
12.
Int J Syst Evol Microbiol ; 69(7): 2023-2027, 2019 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-31066656

RESUMO

A Gram-stain-negative, rod-shaped strain isolated from pig-production environments was identified as a new species within the genus Yersinia using multifaceted genomic and biochemical approaches. The genome of this strain was closed using a hybrid assembly approach combining both high accuracy short read sequencing data with long read sequencing technology. Phylogenetic analysis of the 16S rRNA gene showed ~98 % similarity to Yersinia kristensenii and ~98 % similarity to Yersinia enterocolitica. Average nucleotide identity (OrthoANI) values were calculated as 85.79 % to Y. kristensenii ATCC 33638T and 85.73 % to Y. enterocolitica ATCC 9610T thereby providing evidence that this isolate should be considered as a novel species. The type strain is CFS1934T (=NCTC 14222T=LMG 31076T).


Assuntos
Filogenia , Suínos/microbiologia , Yersinia/classificação , Animais , Técnicas de Tipagem Bacteriana , Composição de Bases , DNA Bacteriano/genética , Irlanda , Tonsila Palatina/microbiologia , RNA Ribossômico 16S/genética , Análise de Sequência de DNA , Yersinia/isolamento & purificação
14.
Mol Cell Probes ; 45: 14-18, 2019 06.
Artigo em Inglês | MEDLINE | ID: mdl-30930280

RESUMO

Actinobacillus pleuropneumoniae is the etiological agent of swine contagious pleuropneumoniae, which is distributed globally and associated with severe economic losses in the pig rearing industry. In this study, a real-time recombinase polymerase amplification assay (real-time RPA) based on the apxIVA gene was developed to rapid detect A. pleuropneumoniae. Real-time RPA was performed successfully in Genie III at the constant temperature of 39 °C for 20 min. The developed assay was highly specific for A. pleuropneumoniae, and the sensitivity at 95% probability was 536 fg of A. pleuropneumoniae genomic DNA. The real-time RPA for A. pleuropneumoniae was further evaluated on the 112 clinical swine lung and tonsil samples, and 25 (22.3%), 27 (24.1%), and 12 (10.7%) samples were positive for A. pleuropneumoniae by the real-time RPA, real-time PCR and bacterial isolation, respectively. With a real-time PCR as the reference method, the real-time RPA showed a diagnostic specificity of 98.8%, a diagnostic sensitivity of 88.9%, a positive predicative value of 96.0%, a negative predictive value of 96.5%, and a kappa value of 0.900. The above data demonstrated the well potentiality and usefulness of the developed real-time RPA assay in the reliable detection of A. pleuropneumoniae, especially in resource limited settings.


Assuntos
Infecções por Actinobacillus/diagnóstico , Actinobacillus pleuropneumoniae/isolamento & purificação , Técnicas de Amplificação de Ácido Nucleico/veterinária , Doenças dos Suínos/virologia , Infecções por Actinobacillus/veterinária , Actinobacillus pleuropneumoniae/genética , Animais , Proteínas de Bactérias/genética , Pulmão/microbiologia , Tonsila Palatina/microbiologia , Reação em Cadeia da Polimerase em Tempo Real/veterinária , Recombinases/metabolismo , Sensibilidade e Especificidade , Suínos
15.
J Clin Microbiol ; 57(7)2019 07.
Artigo em Inglês | MEDLINE | ID: mdl-30944194

RESUMO

Streptococcus suis is one of the most important zoonotic bacterial pathogens of pigs, causing significant economic losses to the global swine industry. S. suis is also a very successful colonizer of mucosal surfaces, and commensal strains can be found in almost all pig populations worldwide, making detection of the S. suis species in asymptomatic carrier herds of little practical value in predicting the likelihood of future clinical relevance. The value of future molecular tools for surveillance and preventative health management lies in the detection of strains that genetically have increased potential to cause disease in presently healthy animals. Here we describe the use of genome-wide association studies to identify genetic markers associated with the observed clinical phenotypes (i) invasive disease and (ii) asymptomatic carriage on the palatine tonsils of pigs on UK farms. Subsequently, we designed a multiplex PCR to target three genetic markers that differentiated 115 S. suis isolates into disease-associated and non-disease-associated groups, that performed with a sensitivity of 0.91, a specificity of 0.79, a negative predictive value of 0.91, and a positive predictive value of 0.79 in comparison to observed clinical phenotypes. We describe evaluation of our pathotyping tool, using an out-of-sample collection of 50 previously uncharacterized S. suis isolates, in comparison to existing methods used to characterize and subtype S. suis isolates. In doing so, we show our pathotyping approach to be a competitive method to characterize S. suis isolates recovered from pigs on UK farms and one that can easily be updated to incorporate global strain collections.


Assuntos
Portador Sadio/veterinária , Infecções Estreptocócicas/veterinária , Streptococcus suis/isolamento & purificação , Streptococcus suis/patogenicidade , Doenças dos Suínos/microbiologia , Animais , Portador Sadio/microbiologia , Inglaterra , Marcadores Genéticos/genética , Genoma Bacteriano/genética , Técnicas de Diagnóstico Molecular , Reação em Cadeia da Polimerase Multiplex , Tonsila Palatina/microbiologia , Infecções Estreptocócicas/microbiologia , Streptococcus suis/genética , Suínos , Virulência/genética , País de Gales
16.
Int J Food Microbiol ; 295: 8-18, 2019 Apr 16.
Artigo em Inglês | MEDLINE | ID: mdl-30776731

RESUMO

Tonsils in the oral cavity are an important source of contamination during pig slaughter, but have not received as much attention as faecal contamination. In the present study, ten pigs were sampled from tonsils, faeces and three different areas on each carcass. The samples were analysed by both culturing of Escherichia coli and Yersinia enterocolitica and by 16S rRNA gene sequencing to characterize the bacterial communities. Comparing culture data from deep tonsil tissue and tonsil surface showed similar numbers of E. coli but significantly higher numbers of Y. enterocolitica in the deep tissue samples. Microbiota analysis showed similar bacterial communities in the two sample types at phylum level, while comparison at genus level showed significant differences between the relative abundance of several genera in the two sample types. The finding of a significantly higher relative abundance of Yersinia in tonsil tissue compared to tonsil surface supported the culture analysis. The microbiota analysis also investigated characteristics of the bacterial community that could discriminate bacterial transfer from tonsils and faeces to the carcass during slaughter. The microbiota analyses demonstrated that Fusobacteria and Proteobacteria are the most abundant phyla in tonsils, while Firmicutes showed the highest relative abundance in faeces. The dominating phylum on carcasses was Proteobacteria. Besides Proteobacteria, the swabbing area on the forepart of the carcass, showed a higher relative abundance of Firmicutes and Fusobacteria compared to swabbing areas on the rear part and mid-section of the carcass. Principal coordinate analysis showed clear clustering of samples based on sample source (tonsils, faeces and carcass). Carcass swab samples from the forepart tended to cluster closer to the tonsil samples compared to carcass swab samples from the rear part and mid-section. Identification of the genera Fusobacterium, Moraxella, Actinobacillus and non-E. coli genera of the family Enterobacteriaceae in carcass swabs could indicate tonsil contamination, while faecal contamination would more likely include higher prevalence of bacteria belonging to the class of Clostridia. The present study supports that it is possible to identify bacterial groups that are indicative for either tonsil or faecal carcass contamination. The level and composition of Enterobacteriaceae on the carcasses did, however, indicate that other sources of meat contamination than tonsils and faeces may be important, such as the process environment.


Assuntos
Fenômenos Fisiológicos Bacterianos , Microbiologia de Alimentos , Carne/microbiologia , Tonsila Palatina/microbiologia , Matadouros , Animais , Bactérias/genética , Fezes/microbiologia , Microbiota/genética , Prevalência , RNA Ribossômico 16S/genética , Suínos
17.
Transbound Emerg Dis ; 66(3): 1218-1226, 2019 May.
Artigo em Inglês | MEDLINE | ID: mdl-30720247

RESUMO

The importance of wild boars as game species in Spain is well known. Their feeding habits and intrusive behaviour, together with a progressively wider spreading of populations, increases the interactions of these animals with livestock and humans. Considering that wild boars could have a potential role in the transmission of certain pathogens as salmonellae, the aims of this study were to determine the prevalence of Salmonella spp. in wild boars hunted in central-western Spain, the occurrence of this pathogen in tonsils, mandibular lymph nodes and faeces (as markers for transmission risk), and to define the phylogenetic relationships among isolated strains, in order to investigate the circulation pathways of bacteria among tissues, animals and estates. Samples from 1,041 hunted wild boars were analysed for the presence of Salmonella spp. by bacteriological culture. Isolates were confirmed by PCR and serotyped in the Spanish national reference laboratory. The genetic relationships between strains were determined by PFGE. The results showed a 7.7% of positive animals (81 wild boars), being tonsils the organ most frequently colonised by Salmonella spp. (18.7%), followed by lymph nodes (5.1%) and faecal samples (2.9%). Serovars Enteritidis and Newport were the most frequent amongst the 34 different serovars obtained. The pulsed-field gel electrophoresis (PGFE) analysis showed a great genetic diversity, with serovars that exhibited different pulsotypes when isolated from different estates and multiple serovars in the same estate. In conclusion, this study reveals the importance of wild boars as carriers and possible transmitters of virulent and/or antimicrobial-resistant clones of Salmonella spp. to livestock and humans.


Assuntos
Fezes/microbiologia , Linfonodos/microbiologia , Tonsila Palatina/microbiologia , Salmonelose Animal/microbiologia , Sus scrofa/microbiologia , Doenças dos Suínos/microbiologia , Animais , Eletroforese em Gel de Campo Pulsado , Humanos , Filogenia , Prevalência , Salmonella/genética , Salmonella/isolamento & purificação , Salmonelose Animal/epidemiologia , Espanha/epidemiologia , Suínos , Doenças dos Suínos/epidemiologia
18.
Int J Pediatr Otorhinolaryngol ; 118: 103-109, 2019 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-30599284

RESUMO

INTRODUCTION: Adenotonsillar and middle ear diseases result in some of the most frequently performed operations in the pediatric population worldwide. The pathogen reservoir hypothesis (PRH) suggests that the adenoids act as a reservoir of bacteria which play a potential pathogenic role in otitis media. Evidence supporting this hypothesis is limited. This study sought to comprehensively determine and compare associations between the adenotonsillar and middle ear bacterial microbiota within individual patients via next-generation sequencing and microbial network analyses. METHODS: Bacterial 16S rRNA gene-targeted amplicon sequencing was used to determine the bacterial composition of ten pediatric patients undergoing adenotonsillectomy and ventilation tube insertion for otitis media with effusion. At the time of surgery, swabs were taken from the adenoid surface, tonsil crypts and middle ear clefts (through the myringotomy incision). RESULTS: The most abundant sequences within the bacterial community at genus level across all anatomical sites were Fusobacterium, Haemophilus, Neisseria, and Porphyromonas. There was an observable difference in the relative abundance of bacterial communities, with a higher proportion of Haemophilus and Moraxella in the adenoid when compared with the middle ear. Furthermore, only one module (consisting of 4 bacterial OTUs) from one patient was identified through microbial network analyses to be significantly associated between middle ear and adenoid. In addition, microbial network analysis revealed that the adenoid and tonsil microbiota share greater similarity than do the adenoid and middle ear. CONCLUSION: The results of this study suggest that the adenoid microenvironment does not correlate to the middle ear microenvironment. A future study at the species level, and over time, is required to further investigate whether the differing relationship between the microbiota of the adenoid and middle ear rejects the pathogen reservoir hypothesis.


Assuntos
Tonsila Faríngea/microbiologia , Bactérias/isolamento & purificação , Orelha Média/microbiologia , Microbiota , Otite Média com Derrame/microbiologia , Tonsila Palatina/microbiologia , Adenoidectomia , Bactérias/genética , Criança , Pré-Escolar , Reservatórios de Doenças/microbiologia , Feminino , Fusobacterium/genética , Fusobacterium/isolamento & purificação , Haemophilus/genética , Haemophilus/isolamento & purificação , Humanos , Masculino , Ventilação da Orelha Média , Moraxella/genética , Moraxella/isolamento & purificação , Neisseria/genética , Neisseria/isolamento & purificação , Otite Média com Derrame/cirurgia , Porphyromonas/genética , Porphyromonas/isolamento & purificação , RNA Ribossômico 16S/análise , Tonsilectomia
19.
PLoS One ; 14(1): e0209975, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-30605453

RESUMO

Mycoplasma hyorhinis and M. hyosynoviae are agents associated with arthritis in pigs. This study investigated the tonsillar detection patterns of M. hyorhinis and M. hyosynoviae in a swine population with a history of lameness. The plausibility of dual PCR detection of these agents in dams at one and three weeks post-farrowing and their offspring at the same time was determined. The association between M. hyorhinis and M. hyosynoviae detection in piglets and potential development of lameness in wean-to-finish stages was evaluated by correlating individual piglet lameness scores and PCR detection in tonsils. Approximately 40% of dams were detected positive for M. hyorhinis and M. hyosynoviae at both one and three weeks post-farrowing. In first parity dams, M. hyorhinis was detected in higher proportions (57.1% and 73.7%) at both weeks of sampling compared to multi-parity dams. A lower proportion of first parity dams (37.5%) were detected positive at week one with M. hyosynoviae and an increase in this proportion to 50% was identified in week three. Only 8.3% of piglets were detected positive for M. hyorhinis in week one compared to week three (50%; p<0.05). The detection of M. hyosynoviae was minimal in piglets at both weeks of sampling (0% and 0.9%). Lameness was scored in pigs 5-22 weeks of age, with the highest score observed at week 5. The correlation between PCR detection and lameness scores revealed that the relative risk of developing lameness post-weaning was significantly associated with detection of M. hyorhinis in piglets at three weeks of age (r = 0.44; p<0.05).The detection pattern of M. hyorhinis and M. hyosynoviae in dams did not reflect the detection pattern in piglets. Results of this study suggest that positive detection of M. hyorhinis in piglets pre-weaning could act as a predictor for lameness development at later production stages.


Assuntos
Infecções por Mycoplasma/veterinária , Mycoplasma hyorhinis/isolamento & purificação , Mycoplasma hyosynoviae/isolamento & purificação , Doenças dos Suínos/microbiologia , Suínos/microbiologia , Animais , Infecções por Mycoplasma/diagnóstico , Infecções por Mycoplasma/microbiologia , Mycoplasma hyorhinis/genética , Mycoplasma hyosynoviae/genética , Tonsila Palatina/microbiologia , Reação em Cadeia da Polimerase , Doenças dos Suínos/diagnóstico , Desmame
20.
Appl Environ Microbiol ; 85(4)2019 02 15.
Artigo em Inglês | MEDLINE | ID: mdl-30530706

RESUMO

Salmonella spp. are estimated to cause 1.2 million cases of human foodborne illness each year in the United States, and pigs can often be asymptomatically colonized with Salmonella spp. (>50% of farms). Recent reports state that 18.3% of Salmonella enterica serovar Typhimurium isolates are resistant to ≥3 antimicrobial classes, and multidrug-resistant (MDR) strains are associated with an increased hospitalization rate and other complications. Chlortetracycline is commonly used in swine production to prevent/treat various diseases; therefore, chlortetracycline treatment of pigs unknowingly colonized with MDR Salmonella may have collateral effects on Salmonella spp. (and other gut bacteria). In this study, we determined the effect of in-feed chlortetracycline (400 g/ton) on shedding and colonization of pigs challenged with the MDR S Typhimurium strain DT104 (n = 11/group). We also assessed the impact on the fecal microbiota over the 12-day experimental period and on the ileum, cecum, and tonsil microbiota at 7 days postinoculation (dpi). In MDR S Typhimurium-inoculated pigs, chlortetracycline administration significantly increased fecal shedding at 2 dpi (+1.4 log10 CFU/g; P < 0.001) and enhanced tonsil colonization (+3.1 log10 CFU/g; P < 0.001). There were few major alterations detected in the gut or tonsillar microbiota of pigs treated with MDR S Typhimurium and/or chlortetracycline. The tonsillar transcriptome was largely unaffected despite increased colonization by MDR S Typhimurium following inoculation of the chlortetracycline-treated pigs. These results highlight the idea that chlortetracycline administration can enhance shedding and colonization of MDR S Typhimurium in pigs, which could increase the risk of environmental dissemination of MDR Salmonella strains.IMPORTANCE Salmonella spp. are an important cause of foodborne illness in North America, and pork products are associated with sporadic cases and outbreaks of human salmonellosis. Isolates of Salmonella may be resistant to multiple antibiotics, and infections with multidrug-resistant (MDR) Salmonella spp. are more difficult to treat, leading to increased hospitalization rates. Swine operations commonly use antimicrobials, such as chlortetracycline, to prevent/treat infections, which may have collateral effects on pig microbial populations. Recently, we demonstrated that chlortetracycline induces the expression of genes associated with pathogenesis and invasion in MDR Salmonella enterica serovar Typhimurium in vitro In our current study, we show increased tonsillar colonization and fecal shedding of the MDR S Typhimurium strain DT104 from pigs administered chlortetracycline. Therefore, pigs unknowingly colonized with multidrug-resistant Salmonella spp. and receiving chlortetracycline for an unrelated infection may be at a greater risk for disseminating MDR Salmonella spp. to other pigs and to humans through environmental or pork product contamination.


Assuntos
Derrame de Bactérias/efeitos dos fármacos , Clortetraciclina/farmacologia , Farmacorresistência Bacteriana Múltipla/efeitos dos fármacos , Fezes/microbiologia , Microbioma Gastrointestinal/efeitos dos fármacos , Tonsila Palatina/microbiologia , Salmonella enterica/efeitos dos fármacos , Ração Animal , Animais , Antibacterianos/farmacologia , Ceco/microbiologia , Salmonelose Animal/tratamento farmacológico , Salmonelose Animal/microbiologia , Salmonelose Animal/prevenção & controle , Sorogrupo , Suínos , Doenças dos Suínos/tratamento farmacológico , Doenças dos Suínos/microbiologia , Doenças dos Suínos/prevenção & controle
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