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1.
Int J Mol Sci ; 22(13)2021 Jun 23.
Artigo em Inglês | MEDLINE | ID: mdl-34201818

RESUMO

Pseudomonas aeruginosa is an opportunistic human pathogen that has become a nosocomial health problem worldwide. The pathogen has multiple drug removal and virulence secretion systems, is resistant to many antibiotics, and there is no commercial vaccine against it. Yersinia pestis is a zoonotic pathogen that is on the Select Agents list. The bacterium is the deadliest pathogen known to humans and antibiotic-resistant strains are appearing naturally. There is no commercial vaccine against the pathogen, either. In the current work, novel compounds based on metallacarborane cage were studied on strains of Pseudomonas aeruginosa and a Yersinia pestis substitute, Yersinia enterocolitica. The representative compounds had IC50 values below 10 µM against Y. enterocolitica and values of 20-50 µM against P. aeruginosa. Artificial generation of compound-resistant Y. enterocolitica suggested a common mechanism for drug resistance, the first reported in the literature, and suggested N-linked metallacarboranes as impervious to cellular mechanisms of resistance generation. SEM analysis of the compound-resistant strains showed that the compounds had a predominantly bacteriostatic effect and blocked bacterial cell division in Y. enterocolitica. The compounds could be a starting point towards novel anti-Yersinia drugs and the strategy presented here proposes a mechanism to bypass any future drug resistance in bacteria.


Assuntos
Antibacterianos/farmacologia , Boranos/química , Compostos Organometálicos/farmacologia , Infecções por Pseudomonas/tratamento farmacológico , Pseudomonas aeruginosa/efeitos dos fármacos , Yersiniose/tratamento farmacológico , Yersinia enterocolitica/efeitos dos fármacos , Humanos , Infecções por Pseudomonas/microbiologia , Yersiniose/microbiologia
2.
J Vet Diagn Invest ; 33(4): 655-663, 2021 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-34075814

RESUMO

The use of oral fluid (OF) to detect zoonotic pathogens in pigs has been only scarcely assessed. We evaluated OF as a potential specimen for detection by culture of methicillin-resistant Staphylococcus aureus (MRSA) and Yersinia enterocolitica, and the detection of antibodies against Salmonella spp. and hepatitis E virus (HEV) using commercial ELISAs. Samples from 33 pig farms were collected at the beginning and end of the fattening period. Results of the OF samples were compared with the results of serum samples and nasal swabs from individual pigs and pen floor fecal samples, using the Cohen kappa (κ) and the McNemar test. For Salmonella spp. antibodies, OF samples were negative, although the corresponding serum samples were positive. The detection of HEV antibodies in sera and OF had agreement at the first sampling, and poor and significant agreement at the second sampling (κ = 0.185, McNemar p = 0.238; κ = 0.088, McNemar p < 0.001). At both sampling times, the detection of MRSA in nasal swabs and OF showed agreement (κ = 0.466, McNemar p = 0.077; κ = 0.603, McNemar p = 1); agreement was seen for the detection of Y. enterocolitica in fecal and OF samples (κ = 0.012, McNemar p = 0.868; κ = 0.082, McNemar p = 0.061, respectively). According to the McNemar test, the use of pen-based OFs is more feasible for the detection of MRSA and Y. enterocolitica by culture than is detection of antibodies by commercial ELISA.


Assuntos
Hepatite E/veterinária , Saliva/microbiologia , Salmonelose Animal/epidemiologia , Infecções Estafilocócicas/veterinária , Doenças dos Suínos/epidemiologia , Yersiniose/veterinária , Animais , Hepatite E/diagnóstico , Hepatite E/epidemiologia , Hepatite E/microbiologia , Vírus da Hepatite E/isolamento & purificação , Staphylococcus aureus Resistente à Meticilina/isolamento & purificação , Prevalência , Salmonella/isolamento & purificação , Salmonelose Animal/diagnóstico , Salmonelose Animal/microbiologia , Estudos Soroepidemiológicos , Infecções Estafilocócicas/diagnóstico , Infecções Estafilocócicas/epidemiologia , Infecções Estafilocócicas/microbiologia , Sus scrofa , Suínos , Doenças dos Suínos/diagnóstico , Doenças dos Suínos/microbiologia , Suíça/epidemiologia , Yersiniose/diagnóstico , Yersiniose/epidemiologia , Yersiniose/microbiologia , Yersinia enterocolitica/isolamento & purificação
3.
Fish Shellfish Immunol ; 110: 55-66, 2021 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-33383177

RESUMO

In this 21-day study, we examined the effects of the aqueous methanolic extract of thin-skinned plum (Prunus domestica) on growth, immune response and resistance to a pathogenic bacterium, Yersinia ruckeri in rainbow trout (Oncorhynchus mykiss). Fish were fed with diets containing thin-skinned plum extract doses as 0 (Control) 0.1 (PD01), 0.5 (PD05) and 1% (PD1) ad libitum twice in a day. At the end of the study, growth was affected positively but not significantly. Feed conversion ratio (FCR) was decreased in the PD01 group (P < 0.05). There were elevated respiratory burst and potential bacterial killing activities on the 7th day in the PD1 fish group. No differences were observed in lysozyme activity (P > 0.05). An increased myeloperoxidase activity was recorded on the 14th day of study. Expression of interleukin and COX-2 genes was elevated on the 7th day of study in the kidney and intestine of treated fish. Histological results indicated no marked changes in organs (gill, kidney, liver and spleen) of PD treated fish groups. Challenge results of fish in all plum extract-treated groups showed an increased survival rate against Y. ruckeri (P < 0.05). This study indicated that the thin-skinned plum aqueous methanolic extract could improve innate immunity, survival against Y. ruckeri and decrease the FCR level.


Assuntos
Adjuvantes Imunológicos/farmacologia , Resistência à Doença , Doenças dos Peixes/prevenção & controle , Oncorhynchus mykiss/imunologia , Extratos Vegetais/farmacologia , Yersinia ruckeri/efeitos dos fármacos , Animais , Doenças dos Peixes/microbiologia , Oncorhynchus mykiss/crescimento & desenvolvimento , Prunus domestica/química , Yersiniose/microbiologia , Yersiniose/prevenção & controle , Yersiniose/veterinária
4.
Food Microbiol ; 94: 103660, 2021 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-33279085

RESUMO

Yersinia enterocolitica bio-serotype 4/O:3 was previously identified in a pork production chain in Brazil and the obtained isolates presented high identity by pulsed-field gel electrophoresis (PFGE, XbaI). For the current study, an additional 147 porcine samples (tonsils = 100, palate = 30, head meat = 17) were collected from the same pork production chain 2-years later and 14 (9.5%) tested positive for Y. enterocolitica. Isolates (n = 24, 1 to 2 per positive sample) were bio-serotype 4/O:3 and harbored virulence genes ail, inv, wbbU, virF, myfA, ystA, ymoA, hreP and sat, and the multidrug resistance related genes emrD, marC and yfhD. PFGE (XbaI) demonstrated no differences among isolates (100% similarity) and were identical to some Y. enterocolitica isolates (n = 13) obtained previously from the same pork chain. A second PFGE analysis (NotI) confirmed the high degree of similarity among isolates obtained over time, demonstrating the persistence of an apparent clonal Y. enterocolitica bio-serotype 4/O:3 in this particular pork production chain in Brazil.


Assuntos
Carne de Porco/microbiologia , Doenças dos Suínos/microbiologia , Yersiniose/veterinária , Yersinia enterocolitica/isolamento & purificação , Animais , Brasil , Eletroforese em Gel de Campo Pulsado , Contaminação de Alimentos/análise , Filogenia , Sorotipagem , Suínos , Yersiniose/microbiologia , Yersinia enterocolitica/classificação , Yersinia enterocolitica/genética
5.
Molecules ; 25(23)2020 Dec 03.
Artigo em Inglês | MEDLINE | ID: mdl-33287412

RESUMO

Lipopolysaccharide (LPS) is the major component of the outer membrane of Gram-negative bacteria, and its integrity is monitored by various stress response systems. Although the Rcs system is involved in the envelope stress response and regulates genes controlling numerous bacterial cell functions of Yersinia enterocolitica, whether it can sense the truncated LPS in Y. enterocolitica remains unclear. In this study, the deletion of the Y. enterocolitica waaF gene truncated the structure of LPS and produced a deep rough LPS. The truncated LPS increased the cell surface hydrophobicity and outer membrane permeability, generating cell envelope stress. The truncated LPS also directly exposed the smooth outer membrane to the external environment and attenuated the resistance to adverse conditions, such as impaired survival under polymyxin B and sodium dodecyl sulfate (SDS) exposure. Further phenotypic experiment and gene expression analysis indicated that the truncated LPS was correlated with the activation of the Rcs phosphorelay, thereby repressing cell motility and biofilm formation. Our findings highlight the importance of LPS integrity in maintaining membrane function and broaden the understanding of Rcs phosphorelay signaling in response to cell envelope stress, thus opening new avenues to develop effective antimicrobial agents for combating Y. enterocolitica infections.


Assuntos
Cápsulas Bacterianas/efeitos dos fármacos , Lipopolissacarídeos/farmacologia , Yersinia enterocolitica/efeitos dos fármacos , Antibacterianos/farmacologia , Cápsulas Bacterianas/metabolismo , Membrana Externa Bacteriana/efeitos dos fármacos , Membrana Externa Bacteriana/metabolismo , Biofilmes/efeitos dos fármacos , Membrana Celular/efeitos dos fármacos , Membrana Celular/metabolismo , Permeabilidade da Membrana Celular/efeitos dos fármacos , Regulação Bacteriana da Expressão Gênica/efeitos dos fármacos , Polimixina B/farmacologia , Transdução de Sinais/efeitos dos fármacos , Dodecilsulfato de Sódio/farmacologia , Yersiniose/tratamento farmacológico , Yersiniose/microbiologia , Yersinia enterocolitica/metabolismo
6.
Vet Microbiol ; 247: 108798, 2020 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-32768239

RESUMO

Yersinia enterocolitica is the most common Yersinia species causing foodborne infections in humans. Pathogenic strains carry the chromosomal ail gene, which is essential for bacterial attachment to and invasion into host cells and for serum resistance. This gene is commonly amplified in several PCR assays detecting pathogenic Y. enterocolitica in food samples and discriminating pathogenic isolates from non-pathogenic ones. We have isolated several non-pathogenic ail-positive Yersinia strains from various sources in Finland. For this study, we selected 16 ail-positive Yersinia strains, which were phenotypically and genotypically characterised. Eleven strains were confirmed to belong to Y. enterocolitica and five strains to Yersinia kristensenii using whole-genome alignment, Parsnp and the SNP phylogenetic tree. All Y. enterocolitica strains belonged to non-pathogenic biotype 1A. We found two copies of the ail gene (ail1 and ail2) in all five Y. kristensenii strains and in one Y. enterocolitica biotype 1A strain. All 16 Yersinia strains carried the ail1 gene consisting of three different sequence patterns (A6-A8), which were highly similar with the ail gene found in high-pathogenic Y. enterocolitica biotype 1B strains (A2). The Ail protein encoded by the ail1 gene was highly conserved compared to the Ail protein encoded by the ail2 gene. Multiple sequence alignment of the ail gene and Ail protein were conducted with MAFF. In total, 10 ail sequence variations have been identified, of which 8 conserved ones belonged to the ail1 gene. According to our results, the detection of ail alone is not sufficient to predict the pathogenicity of Yersinia isolates.


Assuntos
Proteínas da Membrana Bacteriana Externa/genética , Dosagem de Genes , Yersiniose/veterinária , Yersinia enterocolitica/genética , Yersinia/genética , Animais , Finlândia , Genoma Bacteriano , Genótipo , Humanos , Filogenia , Sequenciamento Completo do Genoma , Yersinia/patogenicidade , Yersiniose/microbiologia , Yersinia enterocolitica/patogenicidade
7.
Sci Rep ; 10(1): 11840, 2020 07 16.
Artigo em Inglês | MEDLINE | ID: mdl-32678312

RESUMO

Yersinia ruckeri is an important bacterial pathogen of fish, in particular salmonids, it has been associated with systemic infections worldwide and, like many enteric bacteria, it is a facultative intracellular pathogen. However, the effect of Y. ruckeri's interactions with the host at the cellular level have received little investigation. In the present study, a culture of Chinook Salmon Embryo (CHSE) cell line was exposed to Y. ruckeri. Afterwards, the proteins were investigated and identified by mass spectrometry and compared to the content of unexposed cultures. The results of this comparison showed that 4.7% of the identified proteins were found at significantly altered concentrations following infection. Interestingly, infection with Y. ruckeri was associated with significant changes in the concentration of surface adhesion proteins, including a significantly decreased presence of ß-integrins. These surface adhesion molecules are known to be the target for several adhesion molecules of Yersiniaceae. The concentration of several anti-apoptotic regulators (HSP90 and two DNAj molecules) appeared similarly downregulated. Taken together, these findings suggest that Y. ruckeri affects the proteome of infected cells in a notable manner and our results shed some light on the interaction between this important bacterial pathogen and its host.


Assuntos
Doenças dos Peixes/genética , Proteínas de Peixes/genética , Interações Hospedeiro-Patógeno/genética , Proteoma/genética , Salmão/genética , Yersiniose/genética , Yersinia ruckeri/patogenicidade , Animais , Aderência Bacteriana , Linhagem Celular , Embrião não Mamífero , Doenças dos Peixes/metabolismo , Doenças dos Peixes/microbiologia , Proteínas de Peixes/classificação , Proteínas de Peixes/metabolismo , Perfilação da Expressão Gênica , Regulação da Expressão Gênica , Ontologia Genética , Proteínas de Choque Térmico HSP40/genética , Proteínas de Choque Térmico HSP40/metabolismo , Proteínas de Choque Térmico HSP90/genética , Proteínas de Choque Térmico HSP90/metabolismo , Cadeias beta de Integrinas/genética , Cadeias beta de Integrinas/metabolismo , Anotação de Sequência Molecular , Proteoma/classificação , Proteoma/metabolismo , Salmão/metabolismo , Salmão/microbiologia , Yersiniose/metabolismo , Yersiniose/microbiologia , Yersinia ruckeri/fisiologia
8.
BMC Infect Dis ; 20(1): 498, 2020 Jul 11.
Artigo em Inglês | MEDLINE | ID: mdl-32652941

RESUMO

BACKGROUND: Yersinia enterocolitica is an aero-anaerobic Gram-negative coccobacilli of the Enterobacteriaceae family, rarely reported in osteoarticular infection. CASE PRESENTATION: This report case described a rare septic osteoarticular infection on device due to Yersinia enterocolitica biotype 1B. A purulent fistula appeared after osteosynthesis with plate performed abroad 27 days prior to the presentation for a distal femoral fracture. The treatment consisted of surgical irrigation and washing of the femoral plate and a bitherapy by levoflaxacine and ceftriaxone during 3 months. CONCLUSION: Y. enterocolitica biotype 1B is extremely rare in France. Moreover, the strain implicated in this european case is extremely close from the USA reference strain (with only 2 SNP difference) described in a septicemia in Ohio. The extreme proximity of the strains underlines the need for a sustained surveillance of the spread of this pathogen in France.


Assuntos
Artrite Infecciosa/microbiologia , Placas Ósseas , Infecções Relacionadas à Prótese/microbiologia , Yersiniose/microbiologia , Yersinia enterocolitica/patogenicidade , Idoso de 80 Anos ou mais , Feminino , Fraturas do Fêmur/cirurgia , França , Humanos , Hospedeiro Imunocomprometido , Ohio , Yersinia enterocolitica/genética
9.
Front Immunol ; 11: 988, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32595634

RESUMO

To counteract the deadly pathogens, i.e., Y. pestis, Y. enetrocolitica, and Y. pseudotuberculosis, we prepared a recombinant DNA construct lcrV-hsp70 encoding the bivalent fusion protein LcrV-HSP70. The lcrV gene of Y. pestis and hsp70 domain II DNA fragment of M. tuberculosis were amplified by PCR. The lcrV amplicon was first ligated in the pET vector using NcoI and BamHI restriction sites. Just downstream to the lcrV gene, the hsp70 domain II was ligated using BamHI and Hind III restriction sites. The in-frame and the orientation of cloned lcrV-hsp70 were checked by restriction analysis and nucleotide sequencing. The recombinant bivalent fusion protein LcrV-HSP70 was expressed in E. coli and purified by affinity chromatography. The vaccine potential of LcrV-HSP70 fusion protein was evaluated in formulation with alum. BALB/c mice were vaccinated, and the humoral and cellular immune responses were studied. The fusion protein LcrV-HSP70 induced a strong and significant humoral immune response in comparison to control animals. We also observed a significant difference in the expression levels of IFN-γ and TNF-α in LcrV-HSP70-immunized mice in comparison to control, HSP70, and LcrV groups. To test the protective efficacy of the LcrV-HSP70 fusion protein against plague and Yersiniosis, the vaccinated mice were challenged with Y. pestis, Y. enterocolitica, and Y. pseudotuberculosis separately. The bivalent fusion protein LcrV-HSP70 imparted 100% protection against the plague. In the case of Yersiniosis, on day 2 post challenge, there was a significant reduction in the number of CFU of Y. enterocolitica and Y. pseudotuberculosis in the blood (CFU/ml) and the spleen (CFU/g) of vaccinated animals in comparison to the LcrV, HSP70, and control group animals.


Assuntos
Antígenos de Bactérias/administração & dosagem , Proteínas de Bactérias/administração & dosagem , Vacinas Bacterianas/administração & dosagem , Proteínas de Choque Térmico HSP70/administração & dosagem , Imunogenicidade da Vacina , Proteínas Citotóxicas Formadoras de Poros/administração & dosagem , Vacinação , Vacinas Combinadas/administração & dosagem , Yersiniose/prevenção & controle , Yersinia/imunologia , Animais , Anticorpos Antibacterianos/sangue , Antígenos de Bactérias/genética , Antígenos de Bactérias/imunologia , Carga Bacteriana , Proteínas de Bactérias/genética , Proteínas de Bactérias/imunologia , Vacinas Bacterianas/genética , Vacinas Bacterianas/imunologia , Biomarcadores/sangue , Citocinas/sangue , Feminino , Proteínas de Choque Térmico HSP70/genética , Proteínas de Choque Térmico HSP70/imunologia , Imunidade Celular , Imunidade Humoral , Camundongos Endogâmicos BALB C , Peste , Proteínas Citotóxicas Formadoras de Poros/genética , Proteínas Citotóxicas Formadoras de Poros/imunologia , Proteínas Recombinantes de Fusão/administração & dosagem , Proteínas Recombinantes de Fusão/genética , Proteínas Recombinantes de Fusão/imunologia , Vacinas Combinadas/genética , Vacinas Combinadas/imunologia , Vacinas Sintéticas/administração & dosagem , Vacinas Sintéticas/genética , Vacinas Sintéticas/imunologia , Yersinia/genética , Yersinia/patogenicidade , Yersiniose/imunologia , Yersiniose/microbiologia
10.
Fish Shellfish Immunol ; 103: 472-480, 2020 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-32439514

RESUMO

Recent studies have utilized the fathead minnow (Pimephales promelas) to explore the immunotoxic effects associated with a variety of environmental contaminants in the absence of immunological stimuli. Though this approach allows for alterations in the resting immune system to be detected, previous evidence suggests that many immunotoxic effects may only manifest in the activated immune system. However, basic immune responses to pathogens have not been well described in this species. To expand the utility of the fathead minnow as a model for immunotoxicity testing, a more comprehensive understanding of the activated immune system is required. As such, the main goal of this study was to characterize the transcriptomic response to pathogen infection in the fathead minnow using RNA sequencing. To achieve this goal, female fathead minnows were intraperitoneally injected with either Hank's Balanced Salt Solution (sham-injected) or Yersinia ruckeri (pathogen-injected). Eight hours following injection, fish were sacrificed for the assessment of general morphological (i.e., mass, length, condition factor, hepatic index) and immunological (i.e., leukocyte counts, spleen index) endpoints. To assess the molecular immune response to Y. ruckeri, kidney tissue was collected for transcriptomic analysis. A comparison of sham- and pathogen-injected fish revealed that >1800 genes and >500 gene networks were differentially expressed.Gene networks associated with inflammation, innate immunity, complement, hemorrhaging and iron absorption are highlighted and their utility within the context of immunotoxicity is discussed. These data reveal pathogen-related molecular endpoints to improve data interpretation of future studies utilizing the fathead minnow as a model for immunotoxicity.


Assuntos
Cyprinidae , Doenças dos Peixes/imunologia , Imunidade Inata , Transcriptoma/imunologia , Yersiniose/veterinária , Animais , Feminino , Doenças dos Peixes/microbiologia , Rim/imunologia , Rim/microbiologia , Modelos Animais , Yersiniose/imunologia , Yersiniose/microbiologia , Yersinia ruckeri/fisiologia
11.
Vector Borne Zoonotic Dis ; 20(8): 586-592, 2020 08.
Artigo em Inglês | MEDLINE | ID: mdl-32349633

RESUMO

Rodents are a large group of mammals that can be carriers of zoonotic pathogens such as Yersinia strains that cause yersiniosis. The prevalence of Yersinia enterocolitica and Yersinia pseudotuberculosis was determined in 214 small wild rodents from south-eastern Poland. Samples were analyzed by precultivation and PCR. Nine (4.2%) Y. enterocolitica and one (0.5%) Y. pseudotuberculosis isolates were received. Most of them (n = 5) were obtained from the common vole (Microtus arvalis). All Y. enterocolitica strains were classified as biotype (BT) 1A. A PCR analysis of virulence markers revealed that all Y. enterocolitica isolates contained the ystB gene and five isolates harbored a rare genetic combination of ail/ystB. Three of the four ail/ystB-positive isolates belonged to serotype O:5.27. The Y. pseudotuberculosis inv-positive isolate was classified as BT 1. A genetic analysis of Y. enterocolitica harboring the ystB gene revealed 100% similarity between the analyzed sequences and the sequences from diarrhea patients in India and the United Kingdom as well as high similarity with the sequences from different species of wild animals from Poland. The Y. pseudotuberculosis inv sequence was 100% identical to the sequence isolated from fully virulent clinical strain from France and Australia. The results of our study suggest that small wild rodents, especially voles and yellow-necked mice, may act as carriers of Yersinia strains. The high similarity of the tested gene sequences between our isolates and the isolates from other free-living animals indicates that small wild rodents can play a role in the epidemiology of yersiniosis and can shed Yersinia spp. into the environment.


Assuntos
Doenças dos Roedores/microbiologia , Yersiniose/veterinária , Yersinia enterocolitica/isolamento & purificação , Yersinia pseudotuberculosis/isolamento & purificação , Animais , Animais Selvagens , Polônia/epidemiologia , Prevalência , Doenças dos Roedores/epidemiologia , Roedores , Yersiniose/epidemiologia , Yersiniose/microbiologia
12.
Environ Microbiol ; 22(7): 2939-2955, 2020 07.
Artigo em Inglês | MEDLINE | ID: mdl-32372498

RESUMO

Yersinia ruckeri causes enteric redmouth disease (ERM) that mainly affects salmonid fishes and leads to significant economic losses in the aquaculture industry. An increasing number of outbreaks and the lack of effective vaccines against some serotypes necessitates novel measures to control ERM. Importantly, Y. ruckeri survives in the environment for long periods, presumably by forming biofilms. How the pathogen forms biofilms and which molecular factors are involved in this process, remains unclear. Yersinia ruckeri produces two surface-exposed adhesins, belonging to the inverse autotransporters (IATs), called Y. ruckeri invasin (YrInv) and Y. ruckeri invasin-like molecule (YrIlm). Here, we investigated whether YrInv and YrIlm play a role in biofilm formation and virulence. Functional assays revealed that YrInv and YrIlm promote biofilm formation on different abiotic substrates. Confocal microscopy revealed that they are involved in microcolony interaction and formation, respectively. The effect of both IATs on biofilm formation correlated with the presence of different biopolymers in the biofilm matrix, including extracellular DNA, RNA and proteins. Moreover, YrInv and YrIlm contributed to virulence in the Galleria mellonella infection model. Taken together, we propose that both IATs are possible targets for the development of novel diagnostic and preventative strategies to control ERM.


Assuntos
Doenças dos Peixes/microbiologia , Sistemas de Secreção Tipo V/metabolismo , Virulência/genética , Yersiniose/microbiologia , Yersinia ruckeri/genética , Yersinia ruckeri/patogenicidade , Adesinas Bacterianas , Animais , Biofilmes , Fatores de Virulência/genética , Yersiniose/prevenção & controle
13.
Probiotics Antimicrob Proteins ; 12(4): 1370-1384, 2020 12.
Artigo em Inglês | MEDLINE | ID: mdl-32246325

RESUMO

In the current study, we investigated the effect of a probiotic bacterium (Lactobacillus rhamnosus ATCC 7469) microencapsulated with alginate and hi-maize starch and coated with chitosan on improving growth factors, body composition, blood chemistry, and the immune response of rainbow trout (initial weight: 18.41 ± 0.32 g). Four experimental diets were formulated to feed fish for 60 days. They were control diet without any additive (C), diet added with beads without probiotic (E), a probiotic sprayed to the diet (L.r), and encapsulated probiotic supplemented diet (E-L.r). The results indicated that feeding with E-Lr significantly improved weight gain (84.98 g) and feed conversion ratio (0.95) compared to the other groups (P < 0.05). Also, fish fed E-Lr diet had a significantly higher value of whole-body protein (17.51%), total protein in the blood (4.98 g/dL), lysozyme (30.66 U/mL), alternative complement pathway hemolytic activity (134 U/mL), superoxide dismutase (203 U/mg protein), and catalase (528.33 U/mg protein) (P < 0.05) as compared to those fed the control diet. Similarly, a higher relative expression of immune-related genes such as interleukin-1 (Il-1) and tumor necrosis factor-alpha (TNF-1α) were reported in those fed E-L.r and L.r diets respectively. Interestingly, the fish fed dietary E-L.r had a significantly lower value of lipid in the whole body (4.82%) and cholesterol in the blood (160.67%) in comparison with those fed the control diet (P < 0.05). At the end of the experiment, all groups were challenged by Yersinia ruckeri where the survival rate of rainbow trout fed dietary E-L.r (70.36%) was statistically higher than that of the others (P < 0.05). Overall, the results suggested that encapsulated probiotic Lact. rhamnosus ATCC 7469 acted better than unencapsulated probiotic and has a potential to improve growth performance, flesh quality, and the immune response of rainbow trout.


Assuntos
Doenças dos Peixes/terapia , Proteínas de Peixes/genética , Regulação da Expressão Gênica/efeitos dos fármacos , Lactobacillus rhamnosus/fisiologia , Oncorhynchus mykiss/imunologia , Probióticos/farmacologia , Yersiniose/terapia , Alginatos/química , Ração Animal/análise , Animais , Composição Corporal/efeitos dos fármacos , Catalase/genética , Catalase/imunologia , Encapsulamento de Células/métodos , Células Imobilizadas , Quitosana/química , Colesterol/sangue , Via Alternativa do Complemento/efeitos dos fármacos , Dieta , Resistência à Doença/efeitos dos fármacos , Resistência à Doença/genética , Resistência à Doença/imunologia , Doenças dos Peixes/imunologia , Doenças dos Peixes/microbiologia , Proteínas de Peixes/imunologia , Regulação da Expressão Gênica/imunologia , Interleucina-1/genética , Interleucina-1/imunologia , Muramidase/genética , Muramidase/imunologia , Oncorhynchus mykiss/crescimento & desenvolvimento , Oncorhynchus mykiss/microbiologia , Superóxido Dismutase/genética , Superóxido Dismutase/imunologia , Fator de Necrose Tumoral alfa/genética , Fator de Necrose Tumoral alfa/imunologia , Ganho de Peso/efeitos dos fármacos , Yersiniose/imunologia , Yersiniose/microbiologia , Yersinia ruckeri/efeitos dos fármacos , Yersinia ruckeri/crescimento & desenvolvimento , Yersinia ruckeri/patogenicidade
14.
Sci Rep ; 10(1): 4301, 2020 03 09.
Artigo em Inglês | MEDLINE | ID: mdl-32152417

RESUMO

This study aims to validate the current diagnostic method for the clinical detection of gastroenteritis. We analyzed 400 stool samples to detect three of the most common enteropathogens: Salmonella spp., Campylobacter spp., and Yersinia enterocolitica. All specimens were tested with a routine clinical diagnosis algorithm and with five real-time PCR assays. A total of 98 specimens (24.5%) were positive for enteropathogens. We found 24 samples positive for Salmonella enterica, 71 positive for Campylobacter spp., and 4 positive for Yersinia enterocolitica. All evaluated methods exhibited a good performance in identifying Salmonella and Yersinia enterocolitica, being the highest positive percent agreement (PPA) value of 95.8% and 100%, respectively. The clinical algorithm showed the highest PPA value identifying Salmonella, due to the enrichment in selenite broth. However, the evaluated methods showed notable differences in the identification of Campylobacter species, obtaining a wide range of PPA values: 59.2%-100%. The clinical algorithm showed the lowest PPA value since it was only able to detect Campylobacter jejuni and Campylobacter coli species. This study revealed the importance of implementing the real-time PCR technique in a clinical algorithm: it improved the accuracy of the diagnosis and provided results in a shorter time compared to routine clinical methods.


Assuntos
Algoritmos , Técnicas de Tipagem Bacteriana/métodos , Infecções por Campylobacter/diagnóstico , Fezes/microbiologia , Reação em Cadeia da Polimerase em Tempo Real/métodos , Infecções por Salmonella/diagnóstico , Yersiniose/diagnóstico , Adolescente , Campylobacter/isolamento & purificação , Infecções por Campylobacter/epidemiologia , Infecções por Campylobacter/microbiologia , Criança , Pré-Escolar , Meios de Cultura , Feminino , Humanos , Masculino , Técnicas de Diagnóstico Molecular/métodos , Curva ROC , Salmonella/isolamento & purificação , Infecções por Salmonella/epidemiologia , Infecções por Salmonella/microbiologia , Espanha/epidemiologia , Yersiniose/epidemiologia , Yersiniose/microbiologia , Yersinia enterocolitica/isolamento & purificação
15.
Vet Res ; 51(1): 45, 2020 Mar 20.
Artigo em Inglês | MEDLINE | ID: mdl-32197655

RESUMO

Yersinia ruckeri is a facultative intracellular enterobacterium mostly known as the causative agent of enteric redmouth disease in salmonid fish. In the present study, we applied RNA inhibition to silence twenty pre-selected genes on the genome of a fish cell line (CHSE-214) followed by a gentamicin assay to quantify the effect of silencing on the cells' susceptibility to infection and found that silencing of 18 out of 20 genes significantly reduced the number of Y. ruckeri recovered. These findings improve our understanding of the infection process by Y. ruckeri and of the interactions between this bacterial pathogen and host cells.


Assuntos
Doenças dos Peixes/genética , Proteínas de Peixes/genética , Inativação Gênica , RNA Interferente Pequeno/genética , Yersiniose/veterinária , Animais , Linhagem Celular , Doenças dos Peixes/microbiologia , Proteínas de Peixes/metabolismo , RNA Interferente Pequeno/metabolismo , Yersiniose/genética , Yersiniose/microbiologia , Yersinia ruckeri/fisiologia
16.
J Zoo Wildl Med ; 51(1): 170-176, 2020 Mar 17.
Artigo em Inglês | MEDLINE | ID: mdl-32212560

RESUMO

Yersinia enterocolitica (YE) bioserotype 1B/O:8 (YE 1B/O:8) was identified in routine culture of a variety of zoo species housed at Omaha's Henry Doorly Zoo and Aquarium (OHDZA) from April to July 2011. Animal cases representing 12 species had YE detected from 34 cases during routine fecal monitoring and/or during postmortem examination: Coquerel's sifakas (Propithecus coquereli, two cases), black & white (BW) ruffed lemurs (Varecia variegata variegata, six cases), red ruffed lemurs (Varecia rubra, seven cases), white handed gibbon (Hylobates lar albimana, one case), black lemurs (Eulemur macaco, three cases), mongoose lemurs (Eulemur mongoz, two cases), African hunting dogs (Lycaon pictus, five cases), agile gibbons (Hylobates agilis, three cases), siamangs (Hylobates syndactylus, two cases), colobus monkey (Colobus angolensis palliates, one case), argus pheasant (Argusianus argus, one case), and orangutan (Pongo pygmaeus, one case). Most species were not symptomatic; however, three symptomatic cases in Coquerel's sifakas (two) and a white handed gibbon (one) showed clinical signs of diarrhea and lethargy that resulted in death for the Coquerel's sifakas. One unexpected death also occurred in a BW ruffed lemur. To the authors' knowledge, this is the first report of YE 1B/O:8 in such a large variety of zoo species. The source of the YE could not be identified, prompting the initiation of a diseases surveillance program to prevent further cases for the species that are sensitive to YE. To date, no additional cases have been identified, thus suggesting a single introduction of the YE 1B/O:8 strain into the zoo environment.


Assuntos
Carnívoros , Galliformes , Primatas , Yersiniose/veterinária , Yersinia enterocolitica/fisiologia , Doença Aguda/epidemiologia , Animais , Animais de Zoológico , Derrame de Bactérias , Nebraska/epidemiologia , Sorogrupo , Yersiniose/microbiologia , Yersiniose/mortalidade , Yersiniose/transmissão , Yersinia enterocolitica/genética , Yersinia enterocolitica/isolamento & purificação
17.
Int J Syst Evol Microbiol ; 70(4): 2382-2387, 2020 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-32160143

RESUMO

A Gram-negative rod from the Yersinia genus was isolated from a clinical case of yersiniosis in the United Kingdom. Long read sequencing data from an Oxford Nanopore Technologies (ONT) MinION in conjunction with Illumina HiSeq reads were used to generate a finished quality genome of this strain. Overall Genome Related Index (OGRI) of the strain was used to determine that it was a novel species within Yersinia, despite biochemical similarities to Yersinia enterocolitica. The 16S ribosomal RNA gene accessions are MN434982-MN434987 and the accession number for the complete and closed chromosome is CP043727. The type strain is SRR7544370T (=NCTC 14382T/=LMG 31573T).


Assuntos
Filogenia , Yersiniose/microbiologia , Yersinia/classificação , Técnicas de Tipagem Bacteriana , Composição de Bases , DNA Bacteriano/genética , Genoma Bacteriano , Humanos , RNA Ribossômico 16S/genética , Análise de Sequência de DNA , Espanha , Viagem , Reino Unido , Yersinia/isolamento & purificação
18.
Curr Opin Microbiol ; 54: 111-118, 2020 04.
Artigo em Inglês | MEDLINE | ID: mdl-32092691

RESUMO

Any pathogen worth its salt has mechanisms to evade, subvert, or antagonize host innate immune responses induced by pattern recognition receptors. Resistance against such pathogens therefore requires alternative means to activate protective immune responses. Intriguingly, the receptors that regulate antimicrobial gene expression are coupled to cell death pathways that are activated by blockade of NF-κB and MAPK signaling. In this review, we discuss the regulation of apoptosis in response to pathogen disruption of immune signaling and the role of this cell death response in protection against such pathogens. Stanley often observed that bacterial pathogens are excellent cell biologists and immunologists, and he noted that studying pathogen-host interactions could pave the way to new insights about host biology. Indeed, how Yersinia and other pathogens disrupt innate immune signaling has provided new insight into these pathways and revealed new ways to think about immunogenic properties of apoptosis during bacterial infection.


Assuntos
Infecções Bacterianas/imunologia , Processamento de Proteína Pós-Traducional , Proteína Serina-Treonina Quinases de Interação com Receptores/metabolismo , Yersiniose/imunologia , Yersinia/patogenicidade , Animais , Apoptose , Infecções Bacterianas/microbiologia , Interações Hospedeiro-Patógeno , Humanos , Morte Celular Imunogênica , Camundongos , NF-kappa B/metabolismo , Receptores de Reconhecimento de Padrão/metabolismo , Transdução de Sinais , Yersinia/imunologia , Yersiniose/microbiologia
20.
Infect Immun ; 88(4)2020 03 23.
Artigo em Inglês | MEDLINE | ID: mdl-32014893

RESUMO

Oral administration is a preferred model for studying infection by bacterial enteropathogens such as Yersinia spp. In the mouse model, the most frequent method for oral infection consists of oral gavage with a feeding needle directly introduced in the animal stomach via the esophagus. In this study, we compared needle gavage to bread feeding as an alternative mode of bacterial administration. Using bioluminescence-expressing strains of Yersinia pseudotuberculosis and Yersinia enterocolitica, we detected very early upon needle gavage a bioluminescent signal in the neck area together with a signal in the abdominal region, highlighting the presence of two independent sites of bacterial colonization and multiplication. Bacteria were often detected in the esophagus and trachea, as well as in the lymph nodes draining the salivary glands, suggesting that lesions made during needle introduction into the animal oral cavity lead to rapid bacterial draining to proximal lymph nodes. We then tested an alternative mode of bacterial administration using pieces of bread containing bacteria. Upon bread feeding infection, mice exhibited a stronger bioluminescent signal in the abdominal region than with needle gavage, and no signal was detected in the neck area. Moreover, Y. pseudotuberculosis incorporated in the bread is less susceptible to the acidic environment of the stomach and is therefore more efficient in causing intestinal infections. Based on our observations, bread feeding constitutes a natural and more efficient administration method which does not require specialized skills, is less traumatic for the animal, and results in diseases that more closely mimic foodborne intestinal infection.


Assuntos
Ração Animal , Pão , Modelos Animais de Doenças , Métodos de Alimentação , Gastroenteropatias/microbiologia , Yersiniose/microbiologia , Yersinia enterocolitica/crescimento & desenvolvimento , Yersinia pseudotuberculosis/crescimento & desenvolvimento , Administração Oral , Animais , Camundongos
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