A surveillance of enteropathogens in piglets from birth to seven days of age in Brazil

The purpose of the study was to evaluate the real importance of anaerobic enteropathogens and rotavirus in contrast to more common agents as cause of diarrhea in piglets within the first week of life. Sixty 1- to 7-day-old piglets, 30 diarrheic and 30 non-diarrheic (control), from 15 different herds were selected, euthanized and necropsied. Samples of the jejunum, ileum, colon, cecum and feces were collected from the piglets and analyzed to determine the presence of the following enteropathogens: enterotoxigenic Escherichia coli (ETEC), Clostridium perfringens types A and C, Clostridium difficile, rotavirus and Isospora suis. Among diarrheic piglets, 23.3% were positive for C. difficile, 70% for C. perfringens type A cpb2+, 14.3% for rotavirus and 10% for ETEC. Among non-diarrheic control piglets, 10% were positive for C. difficile, 76.7% for C. perfringens type A cpb2+, 0% for rotavirus, 3.3% for ETEC and 3.3% for I. suis. C. perfringens type C was not detected in any of the animals. Histological lesions characteristic of C. difficile, E. coli and rotavirus were observed. However, no C. perfringens type A suggestive lesions were detected. There was a positive correlation between mesocolon edema and the presence of C. difficile toxins. Although C. perfringens type A cpb2+ was the most frequently detected enteropathogen, there was no association between its presence and diarrhea or macro or microscopic changes. C. difficile and Rotavirus were the most relevant pathogens involved with neonatal diarrhea in this study, and histopathology associated with microbiological test proved to be the key to reach a final diagnosis.

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Deletion of FaeG alleviated Enterotoxigenic Escherichia coli F4ac-induced apoptosis in the intestine

AMB Express ◽

10.1186/s13568-021-01201-z ◽

2021 ◽

Vol 11 (1) ◽

Author(s):

Pengpeng Xia ◽

Yunping Wu ◽

Siqi Lian ◽

Guomei Quan ◽

Yiting Wang ◽

...

Keyword(s):

Escherichia Coli ◽

Clinical Symptoms ◽

Mucosal Damage ◽

Enterotoxigenic Escherichia Coli ◽

Intestinal Epithelial ◽

Antibody Microarray ◽

E Coli ◽

Fimbrial Subunit ◽

Induced Apoptosis ◽

Weaning Piglets

AbstractEnterotoxigenic Escherichia coli (ETEC) F4ac is a major constraint to the development of the pig industry, which is causing newborn and post-weaning piglets diarrhea. Previous studies proved that FaeG is the major fimbrial subunit of F4ac E. coli and efficient for bacterial adherence and receptor recognition. Here we show that the faeG deletion attenuates both the clinical symptoms of F4ac infection and the F4ac-induced intestinal mucosal damage in piglets. Antibody microarray analysis and the detection of mRNA expression using porcine neonatal jejunal IPEC-J2 cells also determined that the absence of FaeG subunit alleviated the F4ac promoted apoptosis in the intestinal epithelial cells. Thus, targeted depletion of FaeG is still beneficial for the prevention or treatment of F4ac infection.

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Multiplex Polymerase Chain Reaction Assay for Identification of Enterotoxigenic Escherichia Coli Strains

Journal of Veterinary Diagnostic Investigation ◽

10.1177/104063870101300405 ◽

2001 ◽

Vol 13 (4) ◽

pp. 308-311 ◽

Cited By ~ 22

Author(s):

Jacek Osek

Keyword(s):

Escherichia Coli ◽

Polymerase Chain Reaction ◽

Multiplex Polymerase Chain Reaction ◽

Direct Determination ◽

Enteric Bacteria ◽

Enterotoxigenic Escherichia Coli ◽

Chain Reaction ◽

Gram Negative ◽

E Coli ◽

Polymerase Chain

A multiplex polymerase chain reaction (PCR) system was developed for identification of enterotoxigenic Escherichia coli (ETEC) strains and to differentiate them from other gram negative enteric bacteria. This test simultaneously amplifies heat-labile (LTI) and heat-stable (STI and STII) toxin sequences and the E. coli-specific universal stress protein ( uspA). The specificity of the method was validated by single PCR tests performed with the reference E. coli and non- E. coli strains and with bacteria isolated from pig feces. The multiplex PCR allowed the rapid and specific identification of enterotoxin-positive E. coli and may be used as a method for direct determination of ETEC and to differentiate them from other E. coli and gram-negative enteric isolates.

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In Vitro and In Vivo Assessment of the Potential of Escherichia coli Phages to Treat Infections and Survive Gastric Conditions

Microorganisms ◽

10.3390/microorganisms9091869 ◽

2021 ◽

Vol 9 (9) ◽

pp. 1869

Author(s):

Joanna Kaczorowska ◽

Eoghan Casey ◽

Gabriele A. Lugli ◽

Marco Ventura ◽

David J. Clarke ◽

...

Keyword(s):

Escherichia Coli ◽

Galleria Mellonella ◽

Enterotoxigenic Escherichia Coli ◽

E Coli ◽

Composite Mixture ◽

Ph Conditions ◽

The Stability ◽

Higher Sensitivity

Enterotoxigenic Escherichia coli (ETEC) and Shigella ssp. infections are associated with high rates of mortality, especially in infants in developing countries. Due to increasing levels of global antibiotic resistance exhibited by many pathogenic organisms, alternative strategies to combat such infections are urgently required. In this study, we evaluated the stability of five coliphages (four Myoviridae and one Siphoviridae phage) over a range of pH conditions and in simulated gastric conditions. The Myoviridae phages were stable across the range of pH 2 to 7, while the Siphoviridae phage, JK16, exhibited higher sensitivity to low pH. A composite mixture of these five phages was tested in vivo in a Galleria mellonella model. The obtained data clearly shows potential in treating E. coli infections prophylactically.

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Detection of Enterotoxigenic Escherichia coli in Rotavirus-Infected Ghanaian Children Diagnosed with Acute Gastroenteritis

American Journal of Tropical Medicine and Hygiene ◽

10.4269/ajtmh.21-0717 ◽

2021 ◽

Author(s):

Bartholomew Dzudzor ◽

Albert Amenyedor ◽

Vincent Amarh ◽

George E. Armah

Keyword(s):

Escherichia Coli ◽

Developing Countries ◽

Enterotoxigenic Escherichia Coli ◽

Surveillance Study ◽

Vaccination Program ◽

E Coli ◽

Rotavirus Vaccination ◽

Stool Samples ◽

Global Health Problem ◽

Vaccine Surveillance

Diarrhea is a notable global health problem in several developing countries, especially in children. Prior to the introduction of the rotavirus vaccination program in Ghana, a surveillance study was conducted to investigate the prevalence of the disease caused by rotavirus in children. In this report, we re-used archival stool samples from the pre-vaccine surveillance study to provide information on prevalence of enterotoxigenic Escherichia coli in Ghanaian children. Re-analysis of the stool samples revealed co-infection of enterotoxigenic E. coli and rotavirus in 2% of the children whose samples were selected for this study. As Ghana is approaching 10 years post-implementation of the rotavirus vaccination program, the preliminary data presented in this report are a vital reference for subsequent studies aimed at ascertaining the effect of the vaccine on both rotavirus and enterotoxigenic E. coli.

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DNA Colony Hybridization Method Using Synthetic Oligonucleotides to Detect Enterotoxigenic Escherichia coli: Collaborative Study

Journal of AOAC INTERNATIONAL ◽

10.1093/jaoac/69.3.531 ◽

1986 ◽

Vol 69 (3) ◽

pp. 531-536

Author(s):

Walter E Hill ◽

Barry A Wentz ◽

William L Payne ◽

James A Jagow ◽

Gerald Zon ◽

...

Keyword(s):

Escherichia Coli ◽

Recombinant Dna ◽

Collaborative Study ◽

Dna Probes ◽

Enterotoxigenic Escherichia Coli ◽

Hybridization Probe ◽

Colony Hybridization ◽

E Coli ◽

Calf Thymus ◽

Recombinant Dna Techniques

Abstract The genes that encode several of the enterotoxins produced by Escherichia coli have been cloned by recombinant DNA techniques. When the nucleotide sequence of these genes is determined, defined sequence oligonucleotides that include a part of these genes may be synthesized. A 22-base DNA hybridization probe was produced for each of 2 heatstable E. coli enterotoxin (ST) genes: STH, from strains originally isolated from humans; and STP, from strains first found in pigs. For this study, 32P end-labeled DNA probes, sonicated calf thymus DNA, and 3 known and 20 unknown (10 ST-positive and 10 ST-negative) strains were sent to each of 23 collaborators. Cultures were spotted onto an agar-based medium and grown into colonies, which were transferred by blotting to cellulose filters, lysed by alkali and steam, and used for DNA colony hybridization with the ST DNA probes. Strains containing an ST gene were recognized as dark spots on an autoradiogram. Of the 460 samples analyzed, 440 (95.7%) were correctly classified by the collaborators. The method has been adopted official first action.

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Enterotoxigenic Escherichia coli infection and intestinal thiamin uptake: studies with intestinal epithelial Caco-2 monolayers

AJP Cell Physiology ◽

10.1152/ajpcell.00276.2013 ◽

2013 ◽

Vol 305 (11) ◽

pp. C1185-C1191 ◽

Cited By ~ 8

Author(s):

Abhisek Ghosal ◽

Nabendu S. Chatterjee ◽

Tristan Chou ◽

Hamid M. Said

Keyword(s):

Escherichia Coli ◽

Significant Inhibition ◽

Water Soluble ◽

Enterotoxigenic Escherichia Coli ◽

Adenylate Cyclase System ◽

Mrna Levels ◽

Intestinal Epithelial ◽

E Coli ◽

Heat Labile ◽

Etec Infection

Infections with enteric pathogens like enterotoxigenic Escherichia coli ( ETEC) is a major health issue worldwide and while diarrhea is the major problem, prolonged, severe, and dual infections with multiple pathogens may also compromise the nutritional status of the infected individuals. There is almost nothing currently known about the effect of ETEC infection on intestinal absorptions of water-soluble vitamins including thiamin. We examined the effect of ETEC infection on intestinal uptake of the thiamin using as a model the human-derived intestinal epithelial Caco-2 cells. The results showed that infecting confluent Caco-2 monolayers with live ETEC (but not with boiled/killed ETEC or nonpathogenic E. coli) or treatment with bacterial culture supernatant led to a significant inhibition in thiamin uptake. This inhibition appears to be caused by a heat-labile and -secreted ETEC component and is mediated via activation of the epithelial adenylate cyclase system. The inhibition in thiamin uptake by ETEC was associated with a significant reduction in expression of human thiamin transporter-1 and -2 (hTHTR1 and hTHTR2) at the protein and mRNA levels as well as in the activity of the SLC19A2 and SLC19A3 promoters. Dual infection of Caco-2 cells with ETEC and EPEC (enteropathogenic E. coli) led to compounded inhibition in intestinal thiamin uptake. These results show for the first time that infection of human intestinal epithelial cells with ETEC causes a significant inhibition in intestinal thiamin uptake. This inhibition is mediated by a secreted heat-labile toxin and is associated with a decrease in the expression of intestinal thiamin transporters.

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Epidemiology of Enterotoxigenic Escherichia coli infection in Minnesota, 2016–2017

Epidemiology and Infection ◽

10.1017/s0950268820001934 ◽

2020 ◽

Vol 148 ◽

Author(s):

S. Buuck ◽

K. Smith ◽

R. C. Fowler ◽

E. Cebelinski ◽

V. Lappi ◽

...

Keyword(s):

Escherichia Coli ◽

Stool Culture ◽

Enterotoxigenic Escherichia Coli ◽

Culture Methods ◽

E Coli ◽

Case Comparison ◽

Culture Independent ◽

The Usa ◽

Escherichia Coli Infection ◽

Traveller’S Diarrhoea

Abstract Enterotoxigenic Escherichia coli (ETEC) is a well-established cause of traveller's diarrhoea and occasional domestic foodborne illness outbreaks in the USA. Although ETEC are not detected by conventional stool culture methods used in clinical laboratories, syndromic culture-independent diagnostic tests (CIDTs) capable of detecting ETEC have become increasingly prevalent in the last decade. This study describes the epidemiology of ETEC infections reported to the Minnesota Department of Health (MDH) during 2016–2017. ETEC-positive stool specimens were submitted to MDH to confirm the presence of ETEC DNA by polymerase chain reaction (PCR). Cases were interviewed to ascertain illness and exposures. Contemporaneous Salmonella cases were used as a comparison group in a case-case comparison analysis of risk factors. Of 222 ETEC-positive specimens received by MDH, 108 (49%) were concordant by PCR. ETEC was the sixth most frequently reported bacterial enteric pathogen among a subset of CIDT-positive specimens. Sixty-nine (64%) laboratory-confirmed cases had an additional pathogen codetected with ETEC, including enteroaggregative E. coli (n = 40) and enteropathogenic E. coli (n = 39). Although travel is a risk factor for ETEC infection, only 43% of cases travelled internationally, providing evidence for ETEC as an underestimated source of domestically acquired enteric illness in the USA.

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Effect of bovine colostrum feeding in comparison with milk replacer and natural feeding on the immune responses and colonisation of enterotoxigenic Escherichia coli in the intestinal tissue of piglets

British Journal Of Nutrition ◽

10.1017/s0007114514003201 ◽

2015 ◽

Vol 113 (6) ◽

pp. 923-934 ◽

Cited By ~ 12

Author(s):

Sugiharto Sugiharto ◽

Ann-Sofie Riis Poulsen ◽

Nuria Canibe ◽

Charlotte Lauridsen

Keyword(s):

Escherichia Coli ◽

Immune System ◽

Bovine Colostrum ◽

Enterotoxigenic Escherichia Coli ◽

Expression Level ◽

Milk Replacer ◽

Intestinal Tissue ◽

Treatment Groups ◽

E Coli ◽

Intestinal Immune System

The present study investigated the effect of feeding bovine colostrum (BC) to piglets in comparison with feeding a milk replacer (MR) and conventional rearing by the sow on the intestinal immune system and number of enterotoxigenic Escherichia coli (ETEC) colonising the intestinal tissue. Piglets (23-d-old) were allocated to one of the following four groups: (1) killed at the beginning of the experiment (Base); (2) separated from the sow and fed BC (BC-fed); (3) separated from the sow and fed a MR (MR-fed); (4) kept with the sow (Sow-Milk). Blood was sampled on days 1 and 8, and faecal samples were collected on days 1, 3, 5 and 8. On day 8, piglets were killed and gastrointestinal digesta and intestinal segments were collected. The frequency of diarrhoea was found to be higher (P≤ 0·019) in MR-fed piglets than in BC-fed and Sow-Milk piglets. Piglets from the MR-fed group had the lowest lactic acid bacteria:haemolytic E. coli ratio (Ptreat= 0·064) in the faeces. The number of E. coli colonising the intestinal tissue was higher (P< 0·001) in piglets from the MR-fed group than in those from the BC-fed and Sow-Milk groups. Piglets from the Sow-Milk group had a higher (P= 0·020) mucosal IgG concentration than those from the MR-fed group, but did not exhibit any difference when compared with piglets from the Base and BC-fed groups. Piglets from the BC-fed group exhibited a reduced (P≤ 0·037) expression level of Toll-like receptor-4 in the intestinal mucosa when compared with those from the MR-fed and Sow-Milk groups. The expression level of IL-2 was higher (P≤ 0·051) in piglets from the MR-fed group than in those from the other treatment groups. In conclusion, feeding BC rather than MR to the piglets reduced the colonisation of intestine by ETEC and modulated the intestinal immune system, whereas no differences were observed in piglets fed BC and conventionally reared by the sows.

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Matched case-control study evaluating the frequency of the main agents associated with neonatal diarrhea in piglets

Pesquisa Veterinária Brasileira ◽

10.1590/s0100-736x2011000600008 ◽

2011 ◽

Vol 31 (6) ◽

pp. 505-510 ◽

Cited By ~ 10

Author(s):

Ricardo T. Lippke ◽

Sandra M. Borowski ◽

Sandra M.T. Marques ◽

Suelen O. Paesi ◽

Laura L. Almeida ◽

...

Keyword(s):

Escherichia Coli ◽

Clostridium Difficile ◽

Clostridium Perfringens ◽

Case Control Study ◽

Type A ◽

Case Control ◽

Latex Agglutination ◽

Fecal Samples ◽

Neonatal Diarrhea ◽

Control Study

A case-control study was carried out in litters of 1 to 7-day-old piglets to identify the main infectious agents involved with neonatal diarrhea in pigs. Fecal samples (n=276) from piglets were collected on pig farms in the State of Rio Grande do Sul, Brazil, from May to September 2007. Litters with diarrhea were considered cases (n=129) and normal litters (n=147) controls. The samples were examined by latex agglutination test, PAGE, conventional isolating techniques, ELISA, PCR, and microscopic methods in order to detect rotavirus, bacterial pathogens (Escherichia coli, Clostridium perfringens type A and C, and Clostridium difficile), and parasites (Coccidian and Cryptosporidium spp.). Outbreaks of diarrhea were not observed during sampling. At least one agent was detected in fecal samples on 25 out of 28 farms (89.3%) and in 16 farms (57.1%) more than one agent was found. The main agents diagnosed were Coccidia (42.86%) and rotavirus (39.29%). The main agents identified in litters with diarrhea were Clostridium difficile (10.6%), Clostridium perfringens type A (8.8%) and rotavirus (7.5%); in control litters, Clostridium difficile (16.6%) and Coccidian (8.5%). Beta hemolytic Escherichia coli and Clostridium perfringens type C were not detected. When compared with controls, no agent was significantly associated with diarrhea in case litters. These findings stress the need for caution in the interpretation of laboratorial diagnosis of mild diarrhea in neonatal pigs, as the sole detection of an agent does not necessarily indicate that it is the cause of the problem.

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Multiplex Real-Time PCR Detection of Heat-Labile and Heat-Stable Toxin Genes in Enterotoxigenic Escherichia coli †

Journal of Food Protection ◽

10.4315/0362-028x-69.2.412 ◽

2006 ◽

Vol 69 (2) ◽

pp. 412-416 ◽

Cited By ~ 21

Author(s):

MICHAEL A. GRANT ◽

JINXIN HU ◽

KAREN C. JINNEMAN

Keyword(s):

Escherichia Coli ◽

Real Time ◽

Real Time Pcr ◽

Membrane Filtration ◽

Enterotoxigenic Escherichia Coli ◽

Pcr Analysis ◽

E Coli ◽

Toxin Genes ◽

Heat Stable ◽

Heat Labile

A multiplex real-time PCR method was developed for detection of heat-labile and heat-stable toxin genes in enterotoxigenic Escherichia coli. Approximately 10 CFU per reaction mixture could be detected in rinsates from produce samples. Several foods representative of varieties previously shown to have caused enterotoxigenic E. coli outbreaks were spiked and enriched for 4 or 6 h. Both heat-labile and heat-stable toxin genes could be detected in the foods tested, with the exception of hot sauce, with threshold cycle values ranging from 25.2 to 41.1. A procedure using membrane filtration which would allow enumeration of the enterotoxigenic E. coli population in a food sample in less than 28 h by real-time PCR analysis of colonies picked from media highly selective for E. coli was also developed.

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