RESUMO
The objective of this field trial was to determine the efficacy of a recombinant toxoid vaccine against Shiga toxin 2e (Stx2e) in piglets suffering from edema disease (ED). Three farms with confirmed ED cases were selected for the field trials. On each farm, a total of 40 4-day-old pigs were randomly allocated to either the vaccinated or unvaccinated group, with 20 pigs per group (10 males and 10 females). A 1.0-mL dose of the recombinant toxoid vaccine was administered intramuscularly to pigs in the vaccinated groups in accordance with the manufacturer's recommendations at 4 d of age. A single 2.0-mL dose of phosphate-buffered saline was administered to unvaccinated pigs at the same age. Clinical signs of ED were observed in 12 piglets in the unvaccinated groups and 7 unvaccinated pigs died as a result of ED out of the total number of piglets on Farms A, B, and C. Vaccination had a positive effect on pig growth performance compared to that of unvaccinated pigs on 2 of the 3 farms. Seroconversion of neutralizing antibodies against Stx2e occurred in 70% of piglets in the vaccinated group on Farm A, 75% of vaccinated piglets on Farm B, and 55% of vaccinated piglets on Farm C, when detectable antibodies were measured at 17 d post-vaccination (dpv). Detectable antibodies were measured in 85% of vaccinated piglets on Farms A and B and in 90% of these piglets on Farm C at 37 dpv. These field trials determined that the ED recombinant toxoid vaccine successfully reduced clinical signs and mortality, improved average daily weight gain, and resulted in the production of neutralizing antibodies against Stx2e in pigs.
L'objectif de cette étude sur le terrain était de déterminer l'efficacité d'un vaccin à base d'anatoxine recombinante contre la toxine Shiga 2e (Stx2e) chez les porcelets souffrant de la maladie de l'oedème (ED). Trois fermes ayant des cas confirmés de ED ont été sélectionnées pour les études sur le terrain. Dans chaque ferme, un total de 40 porcs âgés de 4 jours ont été répartis au hasard dans le groupe vacciné ou non vacciné, avec 20 porcs par groupe (10 mâles et 10 femelles). Une dose de 1,0 ml du vaccin à base d'anatoxine recombinante a été administrée par voie intramusculaire aux porcs des groupes vaccinés conformément aux recommandations du fabricant à l'âge de 4 jours. Une dose unique de 2,0 ml de solution saline tamponnée au phosphate a été administrée aux porcs non vaccinés au même âge. Des signes cliniques de ED ont été observés chez 12 porcelets des groupes non vaccinés et 7 porcs non vaccinés sont morts des suites d'une maladie de l'oedème sur le nombre total de porcelets des fermes A, B et C. La vaccination a eu un effet positif sur les performances de croissance des porcs par rapport à celles des porcs non vaccinés dans 2 des 3 fermes. La séroconversion des anticorps neutralisants contre Stx2e s'est produite chez 70 % des porcelets du groupe vacciné de la ferme A, 75 % des porcelets vaccinés de la ferme B et 55 % des porcelets vaccinés de la ferme C, lorsque des anticorps détectables ont été mesurés 17 jours après la vaccination (dpv). Des anticorps détectables ont été mesurés chez 85 % des porcelets vaccinés des fermes A et B et chez 90 % de ces porcelets de la ferme C à 37 dpv. Ces études sur le terrain ont déterminé que le vaccin toxoïde recombinant ED réduisait avec succès les signes cliniques et la mortalité, améliorait le gain de poids quotidien moyen et entraînait la production d'anticorps neutralisants contre Stx2e chez les porcs.(Traduit par Docteur Serge Messier).
Assuntos
Vacinas Sintéticas , Animais , Suínos , Feminino , Vacinas Sintéticas/imunologia , Vacinas Sintéticas/administração & dosagem , Masculino , Doenças dos Suínos/prevenção & controle , Doenças dos Suínos/imunologia , Edematose Suína/prevenção & controle , Edematose Suína/imunologia , Infecções por Escherichia coli/veterinária , Infecções por Escherichia coli/prevenção & controle , Infecções por Escherichia coli/imunologia , Toxina Shiga II/imunologia , Vacinas contra Escherichia coli/imunologia , Vacinas contra Escherichia coli/administração & dosagem , Toxoides/imunologia , Vacinas Bacterianas/imunologia , Vacinas Bacterianas/administração & dosagemRESUMO
Mammary pathogenic Escherichia coli (MPEC) causes mastitis, which results in substantial economic losses to the dairy industry. A high percentage of Escherichia coli isolated from cows with clinical mastitis harbor adhesin genes, such as fimH. However, it is unclear whether these adhesins are important in the adhesion of MPEC to bovine mammary epithelial cells (BMECs). Therefore, we investigated the effect of adhesins (EcpD, FdeC, and FimH) in MPEC on adherence to the bovine mammary epithelium using cultured BMECs. For this purpose, we used wild-type MPEC as well as single- and double-mutants of fimH, ecpD, and fdeC, and performed adhesion assays with BMECs. First, BMECs were cultured in the presence of lactogenic hormones to induce milk component production and tight junction formation. The bacterial count of the wild-type strain that adhered to the BMECs increased in a dose-dependent manner. In deletion mutant strains, the ΔfimH strain showed lower adhesion (P < 0.05), whereas the adhesion ratio of the ΔecpD and ΔfdeC strains was not statistically different compared with that of the wild-type strain (P > 0.05). Additionally, the fimH/fdeC double-deletion mutants showed the lowest adhesion to BMECs. In conclusion, FimH is crucial in the adhesion of MPEC to BMECs. Overall, our work identifies FimH or FimH/FdeC as interesting targets for future drugs or vaccines to improve the treatment, prevention or chronicity of mastitis induced by MPEC.
Assuntos
Adesinas de Escherichia coli , Aderência Bacteriana , Células Epiteliais , Escherichia coli , Glândulas Mamárias Animais , Animais , Bovinos , Células Epiteliais/microbiologia , Escherichia coli/genética , Adesinas de Escherichia coli/genética , Adesinas de Escherichia coli/metabolismo , Feminino , Glândulas Mamárias Animais/microbiologia , Mastite Bovina/microbiologia , Infecções por Escherichia coli/microbiologia , Proteínas de Fímbrias/genética , Proteínas de Fímbrias/metabolismo , Proteínas de Escherichia coli/genética , Proteínas de Escherichia coli/metabolismoRESUMO
Introduction: Enterotoxigenic Escherichia coli (ETEC) is the main diarrhea-causing pathogen in children and young animals and has become a global health concern. Berberine is a type of "medicine and food homology" and has a long history of use in China, particularly in treating gastrointestinal disorders and bacterial diarrhea. Methods: In this study, we explored the effects of berberine on growth performance, intestinal inflammation, oxidative damage, and intestinal microbiota in a weaned piglet model of ETEC infection. Twenty-four piglets were randomly divided into four groups-a control group (fed a basal diet [BD] and infused with saline), a BD+ETEC group (fed a basal diet and infused with ETEC), a LB+ETEC group (fed a basal diet with 0.05% berberine and infused with ETEC infection), and a HB+ETEC group (fed a basal diet with 0.1% berberine and infused with ETEC). Results: Berberine significantly improved the final body weight (BW), average daily gain (ADG), and average daily feed intake (ADFI) (P<0.05) of piglets, and effectively decreased the incidence of diarrhea among the animals (P<0.05). Additionally, berberine significantly downregulated the expression levels of the genes encoding TNF-α, IL-1ß, IL-6, IL-8, TLR4, MyD88, NF-κB, IKKα, and IKKß in the small intestine of piglets (P<0.05). ETEC infection significantly upregulated the expression of genes coding for Nrf2, CAT, SOD1, GPX1, GST, NQO1, HO-1, GCLC, and GCLM in the small intestine of the animals (P<0.05). Berberine significantly upregulated 12 functional COG categories and 7 KEGG signaling pathways. A correlation analysis showed that berberine significantly increased the relative abundance of beneficial bacteria (Gemmiger, Pediococcus, Levilactobacillus, Clostridium, Lactiplantibacillus, Weissella, Enterococcus, Blautia, and Butyricicoccus) and decreased that of pathogenic bacteria (Prevotella, Streptococcus, Parabacteroides, Flavonifractor, Alloprevotella) known to be closely related to intestinal inflammation and oxidative stress in piglets. In conclusion, ETEC infection disrupted the intestinal microbiota in weaned piglets, upregulating the TLR4/MyD88/NF-κB and Nrf2 signaling pathways, and consequently leading to intestinal inflammation and oxidative stress-induced damage. Discussion: Our data indicated that berberine can optimize intestinal microbiota balance and modulate the TLR4/MyD88/NF-κB and Nrf2 signaling pathways, thus helping to alleviate intestinal inflammation and oxidative damage caused by ETEC infection in weaned piglets.
Assuntos
Berberina , Modelos Animais de Doenças , Escherichia coli Enterotoxigênica , Infecções por Escherichia coli , Microbioma Gastrointestinal , Estresse Oxidativo , Desmame , Animais , Berberina/farmacologia , Berberina/administração & dosagem , Suínos , Estresse Oxidativo/efeitos dos fármacos , Microbioma Gastrointestinal/efeitos dos fármacos , Infecções por Escherichia coli/veterinária , Infecções por Escherichia coli/imunologia , Infecções por Escherichia coli/tratamento farmacológico , Diarreia/veterinária , Diarreia/tratamento farmacológico , Diarreia/microbiologia , Inflamação , Doenças dos Suínos/microbiologia , Doenças dos Suínos/tratamento farmacológicoRESUMO
Shiga-toxin producing Escherichia coli (STEC) O157 is a food-borne pathogen which causes gastrointestinal illness in humans. Ruminants are considered the main reservoir of infection, and STEC exceedance has been associated with heavy rainfall. In September 2022, a large outbreak of STEC O157:H7 was identified in the United Kingdom (UK). A national-level investigation was undertaken to identify the source of the outbreak and inform risk mitigation strategies. Whole genome sequencing (WGS) was used to identify outbreak cases. Overall, 259 cases with illness onset dates between 5 August and 12 October 2022, were confirmed across the UK. Epidemiological investigations supported a UK grown, nationally distributed, short shelf-life food item as the source of the outbreak. Analytical epidemiology and food chain analysis suggested lettuce as the likely vehicle of infection. Food supply chain tracing identified Grower X as the likely implicated producer. Independent of the food chain investigations, a novel geospatial analysis triangulating meteorological, flood risk, animal density and land use data was developed, also identifying Grower X as the likely source. Novel geospatial analysis and One Health approaches are potential tools for upstream data analysis to predict and prevent contamination events before they occur and to support evidence generation in outbreak investigations.
Assuntos
Mudança Climática , Surtos de Doenças , Infecções por Escherichia coli , Escherichia coli O157 , Microbiologia de Alimentos , Doenças Transmitidas por Alimentos , Lactuca , Lactuca/microbiologia , Humanos , Infecções por Escherichia coli/epidemiologia , Infecções por Escherichia coli/microbiologia , Infecções por Escherichia coli/transmissão , Reino Unido/epidemiologia , Escherichia coli O157/isolamento & purificação , Escherichia coli O157/genética , Doenças Transmitidas por Alimentos/epidemiologia , Doenças Transmitidas por Alimentos/microbiologia , Sequenciamento Completo do Genoma , Escherichia coli Shiga Toxigênica/isolamento & purificação , Escherichia coli Shiga Toxigênica/genética , Adulto , Pessoa de Meia-Idade , Feminino , Masculino , Contaminação de Alimentos/análise , Idoso , Animais , Adolescente , CriançaRESUMO
BACKGROUND: Urolithiasis combined with ESBL-producing E. coli is often difficult to control and leads to higher postoperative infection-related complications. This study was aim to explore the efficacy and necessity for early use of carbapenem antibiotics perioperatively in urolithiasis patients with urinary tract infections caused by ESBL-producing E. coli. METHODS: The study included a total of 626 patients who were separated into two groups: Group I (the ESBL-producing E. coli group) and Group II (the non-ESBL-producing E. coli group). Antibiotic susceptibility testing was performed and the two groups induced postoperative infection-related events were recorded. the efficacy of perioperative antibiotics was evaluated. RESULTS: All strains of E. coli in our research were sensitive to Carbapenems antibiotics. In addition to Carbapenems, the resistance rates of ESBL-producing E. coli to 6 other commonly used antibiotics were higher than those of non-ESBL-producing strains. Based on the preoperative antibiotic susceptibility test for the ESBL-producing E. coli group and the qSOFA score, the Carbapenems were more effective than the ß-lactamase inhibitors (p = 0.08), while for the non-ESBL-producing E. coli group, there was no difference in the treatment effects between Carbapenems, ß-lactamase inhibitors, Ceftazidime and Quinolones (p = 0.975). CONCLUSIONS: Carbapenem antibiotics significantly reduced the incidence of postoperative infection-related events compared with other types of antibiotics for ESBL-producing E. coli infections in patient with urolithiasis.
Assuntos
Carbapenêmicos , Infecções por Escherichia coli , Escherichia coli , Urolitíase , beta-Lactamases , Humanos , Carbapenêmicos/uso terapêutico , Escherichia coli/efeitos dos fármacos , Urolitíase/tratamento farmacológico , Feminino , Masculino , Pessoa de Meia-Idade , beta-Lactamases/metabolismo , Infecções por Escherichia coli/tratamento farmacológico , Idoso , Antibacterianos/uso terapêutico , Infecções Urinárias/tratamento farmacológico , Assistência Perioperatória , Adulto , Estudos Retrospectivos , Testes de Sensibilidade Microbiana , Resultado do TratamentoRESUMO
OBJECTIVE: Urinary tract infection is one of the most common extraintestinal infectious diseases encountered in clinics. It affects both genders and all age groups and constitutes a major health issue in clinical practice worldwide. Uropathogens often develop resistance to therapeutic agents, creating a formidable challenge for physicians to treat these infections. The goal of the current review is to provide current information on therapeutic advancements and interventions in the treatment of urinary tract infections. MATERIALS AND METHODS: Databases like MEDLINE, PubMed, and ClinicalTrials.gov were used as search engines to collect the relevant articles, and the required information was extracted. RESULTS: Research data suggest an increasing prevalence of pathogenic strains that are resistant to standard antimicrobial regimens recommended for the treatment of urinary tract infections. Targeted therapies for urinary tract infections, such as mannosides and pilicides, as well as vaccinations against uropathogenic Escherichia coli, have been developed recently. The efficacy of other strategies like iontophoresis, hydrogel-coated catheters, and antibiofilm therapy is also investigated. Clinical trials conducted between 2014 and 2019 show a rise in interest in a variety of therapies, highlighting the need for a thorough strategy to treat urinary tract infections, particularly in populations of women. CONCLUSIONS: Antimicrobial-resistant strains of Escherichia coli are becoming more common in urinary tract infections, which have led to the development of targeted medicines such as mannosides and pilicides, as well as immunizations against the pathogenic Escherichia coli strains. There is continuing research into alternate approaches, such as hydrogel-coated catheters, antibiofilm therapy, and iontophoresis. Clinical trials conducted between 2014 and 2019 showed a rise in interest in these different treatment approaches.
Assuntos
Antibacterianos , Infecções Urinárias , Humanos , Infecções Urinárias/tratamento farmacológico , Infecções Urinárias/microbiologia , Infecções Urinárias/diagnóstico , Antibacterianos/uso terapêutico , Antibacterianos/farmacologia , Infecções por Escherichia coli/tratamento farmacológico , Infecções por Escherichia coli/microbiologiaRESUMO
BACKGROUND: The emergence and dissemination of multidrug resistant (MDR) bacteria pose a severe threat to public health by limiting clinical treatment and prophylactic options. OBJECTIVES: This study investigates the prevalence of Escherichia coli in broilers, their phenotypic antimicrobial resistance (AMR) profiles and the presence of virulence-associated genes (VAGs) and antimicrobial resistance genes (ARGs) using polymerase chain reaction (PCR). MATERIALS AND METHODS: A total of 216 pooled cloacal samples were collected from 1080 broilers across six districts of Bangladesh. Each pooled sample comprised randomly selected cloacal swabs from five birds per farm. E. coli isolates were identified using standard bacteriological approach, followed by biochemical assays and PCR. Antimicrobial susceptibility was assessed using the Kirby-Bauer disc diffusion method, and the presence of ARGs and VAGs was determined via PCR. Five selected isolates were partially sequenced for five VAGs using Sanger sequencing. RESULTS: A total of 177 E. coli isolates (81.94%, 95% confidence interval: 76.24%-86.53%) were identified. The isolates showed the highest resistance to ampicillin (93.79%), followed by tetracycline (91.53%), erythromycin (89.27%) and ciprofloxacin (87%). Conversely, ceftriaxone (80.79%) showed highest susceptibility, followed by gentamicin (37.29%) and neomycin (31.07%). All isolates were MDR, with a multiple antibiotic resistance indexes were <0.3. A significant percentage (16.38%) of E. coli isolates were MDR to five antimicrobial classes and harboured blaTEM, sul1, ere (A), tetA, tetB and tetC genes. The highest prevalent ARGs were blaTEM (88.14%) followed by ere (A) (83.62%) and sul 1 (72.32%). The prevalence of VAGs was astA (56.50%), iucD (31.07%), iss (21.47%), irp2 (15.82%) and cva/cvi (3.39%), respectively. CONCLUSIONS: This study highlights the presence of ARGs contributing to the development of MDR in E. coli carrying VAGs in broilers. Effective monitoring and surveillance of antimicrobial usage in poultry production systems are urgently required to prevent emergence and dissemination of AMR.
Assuntos
Galinhas , Farmacorresistência Bacteriana Múltipla , Infecções por Escherichia coli , Escherichia coli , Doenças das Aves Domésticas , Animais , Bangladesh/epidemiologia , Doenças das Aves Domésticas/microbiologia , Doenças das Aves Domésticas/epidemiologia , Escherichia coli/genética , Escherichia coli/efeitos dos fármacos , Farmacorresistência Bacteriana Múltipla/genética , Infecções por Escherichia coli/veterinária , Infecções por Escherichia coli/epidemiologia , Infecções por Escherichia coli/microbiologia , Prevalência , Virulência/genética , Antibacterianos/farmacologiaRESUMO
Background: In rabbits, renal abscesses (pus-filled sores) are rare and diagnosis remains challenging. Therefore, in this study, we aimed to determine the clinical manifestation and diagnostic tests associated with renal abscess identification in rabbits. Case Description: A four-and-a-half-year-old castrated male Lionhead rabbit with a history of poor appetite and abdominal distension was admitted to the animal hospital. Blood analysis, radiography, ultrasonography, and computed tomography scans revealed a kidney abscess found within the renal parenchyma, with severe loss of the cortex and medulla, extending toward the capsule. Consequently, the rabbit underwent nephrectomy. The enlarged right kidney was surgically removed. Histopathological examination of the affected kidney showed severe necrosis and ischemic zones, atrophy of the renal tubules, and prominent heterophils with mixed inflammatory cell infiltrates. Immunohistochemistry and polymerase chain reaction confirmed Encephalitozoon cuniculi and Escherichia coli infections, respectively. Conclusion: This report provides novel insights into the diagnosis of renal abscesses in Lionhead rabbits.
Assuntos
Abscesso , Encephalitozoon cuniculi , Encefalitozoonose , Infecções por Escherichia coli , Escherichia coli , Animais , Coelhos , Masculino , Encephalitozoon cuniculi/isolamento & purificação , Infecções por Escherichia coli/veterinária , Infecções por Escherichia coli/diagnóstico , Infecções por Escherichia coli/complicações , Encefalitozoonose/veterinária , Encefalitozoonose/diagnóstico , Encefalitozoonose/patologia , Encefalitozoonose/microbiologia , Abscesso/veterinária , Abscesso/diagnóstico , Abscesso/microbiologia , Abscesso/patologia , Escherichia coli/isolamento & purificação , Nefropatias/veterinária , Nefropatias/microbiologia , Nefropatias/diagnóstico , Nefropatias/patologiaRESUMO
The dynamic interplay between intramammary IgG, formation of antigen-IgG complexes and effector immune cell function is essential for immune homeostasis within the bovine mammary gland. We explore how changes in the recognition and binding of anti-LPS IgG to the glycolipid "functional" core in milk from healthy or clinically diagnosed Escherichia coli (E. coli) mastitis cows' controls endotoxin function. In colostrum, we found a varied anti-LPS IgG repertoire and novel soluble LPS/IgG complexes with direct IgG binding to the LPS glycolipid core. These soluble complexes, absent in milk from healthy lactating cows, were evident in cows diagnosed with E. coli mastitis and correlated with endotoxin-driven inflammation. E. coli mastitis milk displayed a proportional reduction in anti-LPS glycolipid core IgG compared to colostrum. Milk IgG extracts showed that only colostrum IgG attenuated LPS induced endotoxin activity. Furthermore, LPS-stimulated reactive oxygen species (ROS) in milk granulocytes was only suppressed by colostrum IgG, while IgG extracts of neither colostrum nor E. coli mastitis milk influenced N-formylmethionine-leucyl-phenylalanine (fMLP)-stimulated ROS in LPS primed granulocytes. Our findings support bovine intramammary IgG diversity in health and in response to E. coli infection generate milk anti-LPS IgG repertoires that coordinate appropriate LPS innate-adaptive immune responses essential for animal health.
Assuntos
Colostro , Infecções por Escherichia coli , Escherichia coli , Glicolipídeos , Imunoglobulina G , Lipopolissacarídeos , Mastite Bovina , Leite , Animais , Bovinos , Feminino , Colostro/imunologia , Colostro/metabolismo , Imunoglobulina G/imunologia , Imunoglobulina G/metabolismo , Mastite Bovina/imunologia , Mastite Bovina/microbiologia , Escherichia coli/imunologia , Lipopolissacarídeos/imunologia , Leite/imunologia , Glicolipídeos/metabolismo , Glicolipídeos/imunologia , Infecções por Escherichia coli/imunologia , Endotoxinas/imunologia , Endotoxinas/metabolismo , Espécies Reativas de Oxigênio/metabolismo , Granulócitos/imunologia , Granulócitos/metabolismo , Ligação Proteica , Glândulas Mamárias Animais/imunologia , Glândulas Mamárias Animais/metabolismoRESUMO
Tigecycline is a last-resort drug used to treat serious infections caused by multidrug-resistant bacteria. tet(X4) is a recently discovered plasmid-mediated tigecycline resistance gene that confers high-level resistance to tigecycline and other tetracyclines. Since the first discovery of tet(X4) in 2019, it has spread rapidly worldwide, and as a consequence, tigecycline has become increasingly ineffective in the clinical treatment of multidrug-resistant infections. In this study, we identified and analyzed tet(X4)-positive Escherichia coli isolates from duck farms in Hunan Province, China. In total, 976 samples were collected from nine duck farms. Antimicrobial susceptibility testing and whole-genome sequencing (WGS) were performed to establish the phenotypes and genotypes of tet(X4)-positive isolates. In addition, the genomic characteristics and transferability of tet(X4) were determined based on bioinformatics analysis and conjugation. We accordingly detected an E. coli strain harboring tet(X4) and seven other resistance genes in duck feces. Multi-locus sequence typing analysis revealed that this isolate belonged to a new clone, and subsequent genetic analysis indicated that tet(X4) was carried in a 4608-bp circular intermediate, flanked by ISVsa3-ORF2-abh elements. Moreover, it exhibited transferability to E. coli C600 with a frequency of 10-5. The detection of tet(X4)-harboring E, coli strains on duck farms enhances our understanding of tigecycline resistance dynamics. The transferable nature of the circular intermediate of tet(X4) contributing to the spread of tigecycline resistance genes poses a substantial threat to healthcare. Consequently, vigilant monitoring and proactive measures are necessary to prevent their spread.
Assuntos
Antibacterianos , Patos , Infecções por Escherichia coli , Escherichia coli , Fazendas , Tigeciclina , Sequenciamento Completo do Genoma , Animais , Antibacterianos/farmacologia , China , Farmacorresistência Bacteriana/genética , Farmacorresistência Bacteriana Múltipla/genética , Patos/microbiologia , Escherichia coli/genética , Escherichia coli/efeitos dos fármacos , Escherichia coli/isolamento & purificação , Infecções por Escherichia coli/veterinária , Infecções por Escherichia coli/microbiologia , Proteínas de Escherichia coli/genética , Fezes/microbiologia , Transferência Genética Horizontal , Genótipo , Testes de Sensibilidade Microbiana , Tipagem de Sequências Multilocus , Plasmídeos/genética , Doenças das Aves Domésticas/microbiologia , Tigeciclina/farmacologiaRESUMO
Background: The emergence of ESBLs producing cephalosporin-resistant Escherichia coli isolates poses a threat to public health. This study aims to decipher the genetic landscape and gain insights into ESBL-producing E. coli strains belonging to the high-risk clone ST410 from pediatric patients. Methods: 29 E. coli ST410 isolates were collected from young children and subjected to antimicrobial susceptibility testing, Whole-genome sequencing (WGS), serotype analysis, MLST, ESBL genes, virulence genes, and plasmid profiling. Results: Antimicrobial susceptibility testing demonstrated a high level of resistance to cephalosporins followed by aminoglycoside, sulfonamide, carbapenem and penicillin group of antibiotics. However, n=20/29 shows MDR phenotype. Phylogenetic group B2 (n=15) dominated, followed by group D (n=7), group A (n=4), and group B1 (n=3). Serotyping analysis identified O1:H7 (n=8), O2:H1 (n=6), O8:H4 (n=5), O16:H5 (n=4), and O25:H4 (n=3). Other serotypes identified included O6:H1, O15:H5, and O18:H7 (n=1 each). The most commonly detected ESBL genes were bla CTX-M, (n=26), followed by bla TEM (n=23), and bla SHV (n=18). Additionally, bla OXA-1 (n=10), bla OXA-48 (n=5), bla KPC-2 (n=3), bla KPC-3 (n=2), bla NDM-1 (n=4), bla NDM-5 (n=1), bla GES-1 (n=2), bla GES-5 (n=1), and bla CYM-1 (n=3). Notable virulence genes identified within the ST410 isolates included fimH (n=29), papC (n=24), hlyA (n=22), and cnf1 (n=18), among others. Diverse plasmids were observed including IncFIS, IncX4, IncFIA, IncCol, IncI2 and IncFIC with transmission frequency ranges from 1.3X10-2 to 2.7X10-3. Conclusion: The ST410 clone exhibited a complex resistance profile, diverse serotypes, the presence of specific resistance genes (ESBL genes), virulence gene repertoire, and diverse plasmids. The bla CTX-M was the most prevalent ESBL gene detected.
Assuntos
Antibacterianos , Infecções por Escherichia coli , Escherichia coli , Testes de Sensibilidade Microbiana , Tipagem de Sequências Multilocus , Filogenia , Plasmídeos , Fatores de Virulência , Sequenciamento Completo do Genoma , beta-Lactamases , Humanos , beta-Lactamases/genética , Escherichia coli/genética , Escherichia coli/efeitos dos fármacos , Escherichia coli/isolamento & purificação , Infecções por Escherichia coli/microbiologia , Infecções por Escherichia coli/epidemiologia , China/epidemiologia , Pré-Escolar , Antibacterianos/farmacologia , Lactente , Fatores de Virulência/genética , Plasmídeos/genética , Sorogrupo , Masculino , Criança , Feminino , Farmacorresistência Bacteriana Múltipla/genética , Sorotipagem , GenótipoRESUMO
Loop-mediated isothermal amplification (LAMP) is a cost-effective, rapid, and highly specific method of replicating nucleic acids. Adding multiple targets into a single LAMP assay to create a multiplex format is highly desirable for clinical applications but has been challenging due to a need to develop specific detection techniques and strict primer design criteria. This study describes the evaluation of a rapid triplex LAMP assay, MAST ISOPLEX®VTEC, for the simultaneous detection of Shiga toxin/verotoxin 1 and 2 (stx1/vt1 and stx2/vt2) genes in verotoxigenic Escherichia coli (E. coli) (VTEC) isolates with inhibition control (IC) synthetic DNA using a single fluorophore-oligonucleotide probe, MAST ISOPLEX®Probes, integrated into the primer set of each target. MAST ISOPLEX®Probes used in the MAST ISOPLEX®VTEC kit produce fluorescent signals as they integrate with reaction products specific to each target, allowing tracking of multiple amplifications in real time using a real-time analyzer. Initial validation on DNA extracts from fecal cultures and synthetic DNA sequences (gBlocks) showed that the MAST ISOPLEX®VTEC kit provides a method for sensitive simultaneous triplex detection in a single assay with a limit of detection (LOD) of less than 100 target copies/assay and 96% and 100% sensitivity and specificity, respectively.
Assuntos
Técnicas de Amplificação de Ácido Nucleico , Técnicas de Amplificação de Ácido Nucleico/métodos , Humanos , Sensibilidade e Especificidade , Toxina Shiga I/genética , Técnicas de Diagnóstico Molecular/métodos , Toxina Shiga II/genética , Limite de Detecção , Escherichia coli Shiga Toxigênica/genética , Escherichia coli Shiga Toxigênica/isolamento & purificação , Infecções por Escherichia coli/microbiologia , Infecções por Escherichia coli/diagnóstico , Kit de Reagentes para DiagnósticoRESUMO
Diarrheagenic Escherichia coli serotypes are associated with various clinical syndromes, yet the precise correlation between serotype and pathotype remains unclear. A major barrier to such studies is the reliance on antisera-based serotyping, which is culture-dependent, low-throughput, and cost-ineffective. We have established a highly multiplex PCR-based serotyping assay, termed the MeltArray E. coli serotyping (EST) assay, capable of identifying 163 O-antigen-encoding genes and 53 H-antigen-encoding genes of E. coli. The assay successfully identified serotypes directly from both simulated and real fecal samples, as demonstrated through spike-in validation experiments and a retrospective study. In a multi-province study involving 637 E. coli strains, it revealed that the five major diarrheagenic pathotypes have distinct serotype compositions. Notably, it differentiated 257 Shigella isolates into four major Shigella species, distinguishing them from enteroinvasive E. coli based on their distinct serotype profiles. The assay's universality was further corroborated by in silico analysis of whole-genome sequences from the EnteroBase. We conclude that the MeltArray EST assay represents a paradigm-shifting tool for molecular serotyping of E. coli, with potential routine applications for comprehensive serotype analysis, disease diagnosis, and outbreak detection.
Assuntos
Infecções por Escherichia coli , Escherichia coli , Fezes , Reação em Cadeia da Polimerase Multiplex , Sorogrupo , Sorotipagem , Sorotipagem/métodos , Infecções por Escherichia coli/microbiologia , Humanos , Escherichia coli/genética , Escherichia coli/classificação , Reação em Cadeia da Polimerase Multiplex/métodos , Fezes/microbiologia , Estudos Retrospectivos , Antígenos O/genética , Diarreia/microbiologia , Shigella/genética , Shigella/classificação , Shigella/isolamento & purificação , Antígenos de Bactérias/genética , Proteínas de Escherichia coli/genéticaRESUMO
AIM: This study aimed to screen and characterize colistin-resistant strains isolated from different livestock species in Algeria, including sheep, goats, and dromedaries. METHODS AND RESULTS: A total of 197 rectal and nasal swabs were screened for colistin-resistant Gram-negative bacilli. Twenty one isolates were selected, identified, and their antibiotic resistance was phenotypically and genotypically characterized. The majority (15/21) were affiliated to Escherichia coli, from which 4 strains isolated from sheep (n = 2) and goats (n = 2) and belonging to phylogroup A and ST10 and ST6396 lineages, respectively, carried the mcr-1 gene. The remaining isolates were identified as belonging to the following genera: Raoultella, Enterobacter, Klebsiella, and Pseudomonas. CONCLUSION: This study highlights the presence of virulent and multiresistant Gram-negative bacilli in farm animals, increasing the risk of transmitting potentially fatal infections to humans.
Assuntos
Antibacterianos , Colistina , Infecções por Escherichia coli , Proteínas de Escherichia coli , Cabras , Animais , Colistina/farmacologia , Ovinos , Antibacterianos/farmacologia , Proteínas de Escherichia coli/genética , Argélia , Infecções por Escherichia coli/veterinária , Infecções por Escherichia coli/microbiologia , Testes de Sensibilidade Microbiana , Escherichia coli/genética , Escherichia coli/isolamento & purificação , Escherichia coli/efeitos dos fármacos , Escherichia coli Enterotoxigênica/genética , Escherichia coli Enterotoxigênica/efeitos dos fármacos , Escherichia coli Enterotoxigênica/isolamento & purificação , Reservatórios de Doenças/microbiologia , Farmacorresistência Bacteriana/genética , Farmacorresistência Bacteriana Múltipla/genética , África do NorteRESUMO
Enterotoxigenic Escherichia coli (ETEC) are a major cause of diarrheal illness in humans and animals, induced by enterotoxins produced by these pathogens. Despite the crucial role of neutrophils in combatting bacterial infections, our understanding of how enterotoxins impact neutrophil function is limited. To address this knowledge gap, we used heat-labile enterotoxin (LT) and heat-stable enterotoxin a (STa) to investigate their impact on the effector functions of neutrophils. Our study reveals that pSTa does not exert any discernible effect on the function of neutrophils. In contrast, LT altered the migration and phagocytosis of neutrophils and induced the production of inflammatory factors via activation of cAMP/PKA and ERK1/2 signaling. LT also attenuated the release of neutrophil extracellular traps by neutrophils via the PKA signaling pathway. Our findings provide novel insights into the impact of LT on neutrophil function, shedding light on the underlying mechanisms that govern its immunoregulatory effects. This might help ETEC in subverting the immune system and establishing infection.
Assuntos
Toxinas Bacterianas , Proteínas Quinases Dependentes de AMP Cíclico , AMP Cíclico , Escherichia coli Enterotoxigênica , Enterotoxinas , Infecções por Escherichia coli , Proteínas de Escherichia coli , Neutrófilos , Fagocitose , Enterotoxinas/metabolismo , Neutrófilos/imunologia , Neutrófilos/metabolismo , Humanos , AMP Cíclico/metabolismo , Proteínas de Escherichia coli/metabolismo , Proteínas de Escherichia coli/genética , Proteínas Quinases Dependentes de AMP Cíclico/metabolismo , Toxinas Bacterianas/metabolismo , Toxinas Bacterianas/imunologia , Infecções por Escherichia coli/imunologia , Infecções por Escherichia coli/microbiologia , Sistema de Sinalização das MAP Quinases/efeitos dos fármacos , Armadilhas Extracelulares/metabolismo , Armadilhas Extracelulares/imunologia , Transdução de SinaisRESUMO
BACKGROUND: In the present study, we aimed to determine the frequency of the csgA, fimH, mrkD, foc, papaGI, papGII and papGIII genes, to provide and to design fimbrial adhesin gene (FAG) patterns and profiles for the isolated uropathogenic Escherichia coli (UPEC) strains. METHODS: The enrollment of 108 positive urine samples was performed during seven months, between January 2022 and July 2022. The UPEC strains were confirmed through the standard microbiological and biochemical tests. The antimicrobial susceptibility test was performed through the Kirby-Bauer disc diffusion method. Molecular screening of FAGs was done through the polymerase chain reaction technology. The statistical analyses including chi square and Fisher's exact tests were performed to interpret the obtained results in the present study. RESULTS: As the main results, the antimicrobial resistance (AMR) patterns, multi- (MDR) and extensively drug-resistance (XDR) patterns and FAG patterns were designed and provided. fimH (93.3%), csgA (90.4%) and papG (37.5%) (papGII (30.8%)) genes were recognized as the top three FAGs, respectively. Moreover, the frequency of csgA-fimH gene profile was identified as the top FAG pattern (46.2%) among the others. The isolates bearing csgA-fimH gene profile were armed with a versatile of phenotypic AMR patterns. In the current study, 27.8%, 69.4% and 1.9% of the UPEC isolates were detected as extended-spectrum ß-lactamases (ESBLs) producers, MDR and XDR strains, respectively. CONCLUSIONS: In conclusion, detection, providing and designing of patterns and profiles in association with FAGs, AMR feature in UPEC strains give us an effective option to have a successful and influential prevention for both of UTIs initiation and AMR feature.
Assuntos
Infecções por Escherichia coli , Proteínas de Escherichia coli , Proteínas de Fímbrias , Fímbrias Bacterianas , Infecções Urinárias , Escherichia coli Uropatogênica , Escherichia coli Uropatogênica/genética , Escherichia coli Uropatogênica/efeitos dos fármacos , Humanos , Proteínas de Escherichia coli/genética , Infecções por Escherichia coli/microbiologia , Infecções Urinárias/microbiologia , Proteínas de Fímbrias/genética , Proteínas de Fímbrias/metabolismo , Fímbrias Bacterianas/genética , Fímbrias Bacterianas/metabolismo , Antibacterianos/farmacologia , Testes de Sensibilidade Microbiana , Adesinas de Escherichia coli/genética , Adesinas de Escherichia coli/metabolismo , Feminino , Adulto , Chaperonas Moleculares/genética , Chaperonas Moleculares/metabolismo , Masculino , Farmacorresistência Bacteriana Múltipla/genética , Pessoa de Meia-Idade , Adulto Jovem , Adolescente , Proteínas de BactériasRESUMO
BACKGROUND: Recurrent bacterial cystitis, often referred to as recurrent urinary tract infection (UTI), can be difficult to manage and alternative treatments are needed. HYPOTHESIS/OBJECTIVE: Intravesicular administration of asymptomatic bacteriuria (ASB) E. coli 212 will not be inferior to antimicrobial treatment for the management of recurrent UTI in dogs. ANIMALS: Thirty-four dogs with >1 UTI in the 12 months before presentation. METHODS: All dogs were deemed normal otherwise based on absence of abnormalities on physical examination, CBC, serum biochemical panel, and abdominal ultrasonography. Dogs were randomized to 1 of 2 treatment groups: Group 1 antimicrobials for 7 days or group 2 intravesicular administration of ASB E. coli 212. Owners were provided a voiding questionnaire regarding their dogs' clinical signs, which was completed daily for 14 days to assess clinical cure. Dogs were examined on days 7 and 14 to assess clinical cure, and urine specimens were submitted for urinalysis and bacterial culture. RESULTS: Clinical cure rates for ASB E. coli 212-treated dogs were not inferior to 7 days of antimicrobial treatment with a 12% margin of difference to determine non-inferiority. No significant difference was found between the treatment groups on days 7 and 14 in the proportion of dogs achieving ≥50% or ≥75% reduction in their clinical score compared with baseline. CONCLUSIONS AND CLINICAL IMPORTANCE: These data suggest that intravesicular administration of ASB E. coli 212 is not inferior to antimicrobials for the treatment of recurrent UTI in dogs. This biotherapeutic agent could help alleviate the need for antimicrobials for some dogs with recurrent UTI, improving antimicrobial stewardship.
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Antibacterianos , Bacteriúria , Doenças do Cão , Infecções por Escherichia coli , Escherichia coli , Animais , Cães , Doenças do Cão/tratamento farmacológico , Escherichia coli/efeitos dos fármacos , Bacteriúria/veterinária , Bacteriúria/tratamento farmacológico , Feminino , Antibacterianos/uso terapêutico , Antibacterianos/administração & dosagem , Masculino , Infecções por Escherichia coli/veterinária , Infecções por Escherichia coli/tratamento farmacológico , Infecções Urinárias/veterinária , Infecções Urinárias/tratamento farmacológicoRESUMO
Background and Objectives: Carbapenem resistance is a growing global challenge for healthcare, and, therefore, monitoring its prevalence and patterns is crucial for implementing targeted interventions to mitigate its impact on patient outcomes and public health. This study aimed to determine the prevalence of carbapenem resistance among Escherichia coli (E. coli) strains in the largest tertiary care hospital of the capital territory of Pakistan and to characterize the isolates for the presence of antimicrobial resistance genes. Additionally, the most prevalent sequence types were analyzed. Materials and Methods: A total of 15,467 clinical samples were collected from November 2020 to May 2022, underwent antimicrobial susceptibility testing, and were analyzed for antimicrobial resistance genes through conventional PCR and sequence typing using MLST. Results: In carbapenem-resistant E. coli (CR-EC), 74.19% of isolates harbored the blaNDM gene, with blaNDM-1 (66.96%), blaNDM-5 (12.17%), and blaNDM-7 (20.87%) variants detected. Additionally, blaIMP was found in 25.81% and blaOXA-48 in 35.48% of isolates. The presence of blaCTX-M15 and blaTEM was identified in 83.87% and 73.55% of CR-EC isolates, respectively, while armA and rmtB were detected in 40% and 65.16% of isolates, respectively. Colistin and tigecycline were the most effective drugs against CR-EC isolates, with both showing an MIC50 of 0.5 µg/mL. The MIC90 for colistin was 1 µg/mL, while for tigecycline, it was 2 µg/mL. MLST analysis revealed that the CR-EC isolates belonged to ST131 (24.52%), ST2279 (23.87%), ST3499 (16.13%), ST8051 (15.48%), ST8900 (9.68%), ST3329 (7.10%), ST88 (1.94%), and ST6293 (1.29%). The ST131 complex (70.97%) was the most prevalent, harboring 95.65% of the blaNDM gene, while the ST23 complex (18.06%) harbored 62.50% of the blaIMP gene. Conclusions: Implementing large-scale surveillance studies to monitor the spread of specific pathogens, along with active infection control policies, is crucial for the effective containment and prevention of future epidemics.
Assuntos
Antibacterianos , Escherichia coli , Hospitais Universitários , Testes de Sensibilidade Microbiana , beta-Lactamases , Escherichia coli/efeitos dos fármacos , Escherichia coli/genética , Humanos , Paquistão/epidemiologia , beta-Lactamases/genética , Antibacterianos/farmacologia , Antibacterianos/uso terapêutico , Infecções por Escherichia coli/epidemiologia , Infecções por Escherichia coli/microbiologia , Infecções por Escherichia coli/tratamento farmacológico , Tipagem de Sequências Multilocus/métodos , Carbapenêmicos/farmacologia , Carbapenêmicos/uso terapêutico , PrevalênciaRESUMO
Urinary tract infections (UTIs) represent a clinical and epidemiological problem of worldwide impact that affects the economy and the emotional state of the patient. Control of the condition is complicated due to multidrug resistance of pathogens associated with the disease. Considering the difficulty in carrying out effective treatment with antimicrobials, it is necessary to propose alternatives that improve the clinical status of the patients. With this purpose, in a previous study, the safety and immunostimulant capacity of a polyvalent lysate designated UNAM-HIMFG prepared with different bacteria isolated during a prospective study of chronic urinary tract infection (CUTI) was evaluated. In this work, using an animal model, results are presented on the immunostimulant and protective activity of the polyvalent UNAM-HIMFG lysate to define its potential use in the control and treatment of CUTI. Female Balb/c mice were infected through the urethra with Escherichia coli CFT073 (UPEC O6:K2:H1) strain; urine samples were collected before the infection and every week for up to 60 days. Once the animals were colonized, sublingual doses of UNAM-HIMFG lysate were administrated. The colonization of the bladder and kidneys was evaluated by culture, and their alterations were assessed using histopathological analysis. On the other hand, the immunostimulant activity of the compound was analyzed by qPCR of spleen mRNA. Uninfected animals receiving UNAM-HIMFG lysate and infected animals administered with the physiological saline solution were used as controls. During this study, the clinical status and evolution of the animals were evaluated. At ninety-six hours after infection, the presence of CFT073 was identified in the urine of infected animals, and then, sublingual administration of UNAM-HIMFG lysate was started every week for 60 days. The urine culture of mice treated with UNAM-HIMFG lysate showed the presence of bacteria for three weeks post-treatment; in contrast, in the untreated animals, positive cultures were observed until the 60th day of this study. The histological analysis of bladder samples from untreated animals showed the presence of chronic inflammation and bacteria in the submucosa, while tissues from mice treated with UNAM-HIMFG lysate did not show alterations. The same analysis of kidney samples of the two groups (treated and untreated) did not present alterations. Immunostimulant activity assays of UNAM-HIMFG lysate showed overexpression of TNF-α and IL-10. Results suggest that the lysate activates the expression of cytokines that inhibit the growth of inoculated bacteria and control the inflammation responsible for tissue damage. In conclusion, UNAM-HIMFG lysate is effective for the treatment and control of CUTIs without the use of antimicrobials.
Assuntos
Infecções por Escherichia coli , Camundongos Endogâmicos BALB C , Bexiga Urinária , Infecções Urinárias , Escherichia coli Uropatogênica , Animais , Infecções Urinárias/microbiologia , Infecções Urinárias/imunologia , Feminino , Camundongos , Bexiga Urinária/microbiologia , Bexiga Urinária/imunologia , Bexiga Urinária/patologia , Bexiga Urinária/efeitos dos fármacos , Infecções por Escherichia coli/imunologia , Infecções por Escherichia coli/microbiologia , Escherichia coli Uropatogênica/imunologia , Escherichia coli Uropatogênica/patogenicidade , Modelos Animais de Doenças , Adjuvantes Imunológicos/farmacologia , Lisados BacterianosRESUMO
Adhesive-invasive E. coli has been suggested to be associated with the development of Crohn's disease (CD). It is assumed that they can provoke the onset of the inflammatory process as a result of the invasion of intestinal epithelial cells and then, due to survival inside macrophages and dendritic cells, stimulate chronic inflammation. In previous reports, we have shown that passage of the CD isolate ZvL2 on minimal medium M9 supplemented with sodium propionate (PA) as a carbon source stimulates and inhibits the adherent-invasive properties and the ability to survive in macrophages. This effect was reversible and not observed for the laboratory strain K12 MG1655. We were able to compare the isogenic strain AIEC in two phenotypes-virulent (ZvL2-PA) and non-virulent (ZvL2-GLU). Unlike ZvL2-GLU, ZvL2-PA activates the production of ROS and cytokines when interacting with neutrophils. The laboratory strain does not cause a similar effect. To activate neutrophils, bacterial opsonization is necessary. Differences in neutrophil NADH oxidase activation and ζ-potential for ZvL2-GLU and ZvL2-PA are associated with changes in membrane protein abundance, as demonstrated by differential 2D electrophoresis and LC-MS. The increase in ROS and cytokine production during the interaction of ZvL2-PA with neutrophils is associated with a rearrangement of the abundance of membrane proteins, which leads to the activation of Rcs and PhoP/Q signaling pathways and changes in the composition and/or modification of LPS. Certain isoforms of OmpA may play a role in the formation of the virulent phenotype of ZvL2-PA and participate in the activation of NADPH oxidase in neutrophils.