Quantification of viable but nonculturable Salmonella spp. and Shigella spp. during sludge anaerobic digestion and their reactivation during cake storage.

Science.gov (United States)

Fu, B; Jiang, Q; Liu, H-B; Liu, H

2015-10-01

The presence of viable but nonculturable (VBNC) bacterial pathogens which often fail to be detected by cultivation and can regain the cultivability if the living conditions improve were reported. The objective of this study was to determine the occurrence of VBNC Salmonella spp. and Shigella spp. in the biosolids during anaerobic digestion and its reactivation during the cake storage. The occurrence of VBNC Salmonella spp. and Shigella spp. during mesophilic, temperature-phased, thermophilic anaerobic digestion of sewage sludge and the subsequent storage were studied by RT-qPCR and most probable number (MPN) method. The VBNC incidence of Salmonella spp. and Shigella spp. during thermophilic digestion was four orders of magnitude higher than those of mesophilic digestion. Accordingly, higher resuscitation ratio of VBNC pathogens was also achieved in thermophilic digested sludge. As a result, the culturable Salmonella typhimurium contents in thermophilic digested sludge after cake storage were two orders of magnitude higher than mesophilic digestion. Both quantitative PCR and reverse transcription quantitative PCR assay results showed the two bacterial counting numbers remained stable throughout the cake storage. The results indicate that the increase in the culturable Salmonella spp. and Shigella spp. after centrifugal dewatering was attributed to the resuscitation from the VBNC state to the culturable state. Thermophilic anaerobic digestion mainly induced Salmonella spp. and Shigella spp. into VBNC state rather than killed them, suggesting that the biological safety of sewage sludge by temperature-phased anaerobic digestion should be carefully assessed. © 2015 The Society for Applied Microbiology.

Enteropathogenic Escherichia coli, Samonella, Shigella and Yersinia: cellular aspects of host-bacteria interactions in enteric diseases

Directory of Open Access Journals (Sweden)

Reis Roberta

2010-07-01

Full Text Available Abstract A successful infection of the human intestine by enteropathogenic bacteria depends on the ability of bacteria to attach and colonize the intestinal epithelium and, in some cases, to invade the host cell, survive intracellularly and disseminate from cell to cell. To accomplish these processes bacteria have evolved an arsenal of molecules that are mostly secreted by dedicated type III secretion systems, and that interact with the host, subverting normal cellular functions. Here we overview the most important molecular strategies developed by enteropathogenic Escherichia coli, Salmonella enterica, Shigella flexneri, and Yersinia enterocolitica to cause enteric infections. Despite having evolved different effectors, these four microorganisms share common host cellular targets.

Pre-Clinical Testing of Real-Time PCR Assays for Diarrheal Disease Agents of Genera Escherichia and Shigella

Science.gov (United States)

2014-05-16

FOR DIARRHEAL DISEASE AGENTS OF GENERA ESCHERICHIA AND SHIGELLA May 16, 2014 Reporting Period: October 1, 2010 to September 30, 2013...10-2010 - 30-09-2013 PRE-CLINICAL TESTING OF REAL-TIME PCR ASSAYS FOR DIARRHEAL DISEASE AGENTS OF GENERA ESCHERICHIA AND SHIGELLA ...Texas (MOA 2007 - 2013. Agreement No.: DODI 4000.19; AFI 25-201). Pre-clinical test results qualify ETEC and Shigella real-time PCR assays as lead

Identification of Escherichia coli and Shigella Species from Whole-Genome Sequences.

Science.gov (United States)

Chattaway, Marie A; Schaefer, Ulf; Tewolde, Rediat; Dallman, Timothy J; Jenkins, Claire

2017-02-01

Escherichia coli and Shigella species are closely related and genetically constitute the same species. Differentiating between these two pathogens and accurately identifying the four species of Shigella are therefore challenging. The organism-specific bioinformatics whole-genome sequencing (WGS) typing pipelines at Public Health England are dependent on the initial identification of the bacterial species by use of a kmer-based approach. Of the 1,982 Escherichia coli and Shigella sp. isolates analyzed in this study, 1,957 (98.4%) had concordant results by both traditional biochemistry and serology (TB&S) and the kmer identification (ID) derived from the WGS data. Of the 25 mismatches identified, 10 were enteroinvasive E. coli isolates that were misidentified as Shigella flexneri or S. boydii by the kmer ID, and 8 were S. flexneri isolates misidentified by TB&S as S. boydii due to nonfunctional S. flexneri O antigen biosynthesis genes. Analysis of the population structure based on multilocus sequence typing (MLST) data derived from the WGS data showed that the remaining discrepant results belonged to clonal complex 288 (CC288), comprising both S. boydii and S. dysenteriae strains. Mismatches between the TB&S and kmer ID results were explained by the close phylogenetic relationship between the two species and were resolved with reference to the MLST data. Shigella can be differentiated from E. coli and accurately identified to the species level by use of kmer comparisons and MLST. Analysis of the WGS data provided explanations for the discordant results between TB&S and WGS data, revealed the true phylogenetic relationships between different species of Shigella, and identified emerging pathoadapted lineages. © Crown copyright 2017.

The common structural architecture of Shigella flexneri and Salmonella typhimurium type three secretion needles.

Directory of Open Access Journals (Sweden)

Jean-Philippe Demers

2013-03-01

Full Text Available The Type Three Secretion System (T3SS, or injectisome, is a macromolecular infection machinery present in many pathogenic Gram-negative bacteria. It consists of a basal body, anchored in both bacterial membranes, and a hollow needle through which effector proteins are delivered into the target host cell. Two different architectures of the T3SS needle have been previously proposed. First, an atomic model of the Salmonella typhimurium needle was generated from solid-state NMR data. The needle subunit protein, PrgI, comprises a rigid-extended N-terminal segment and a helix-loop-helix motif with the N-terminus located on the outside face of the needle. Second, a model of the Shigella flexneri needle was generated from a high-resolution 7.7-Å cryo-electron microscopy density map. The subunit protein, MxiH, contains an N-terminal α-helix, a loop, another α-helix, a 14-residue-long β-hairpin (Q51-Q64 and a C-terminal α-helix, with the N-terminus facing inward to the lumen of the needle. In the current study, we carried out solid-state NMR measurements of wild-type Shigella flexneri needles polymerized in vitro and identified the following secondary structure elements for MxiH: a rigid-extended N-terminal segment (S2-T11, an α-helix (L12-A38, a loop (E39-P44 and a C-terminal α-helix (Q45-R83. Using immunogold labeling in vitro and in vivo on functional needles, we located the N-terminus of MxiH subunits on the exterior of the assembly, consistent with evolutionary sequence conservation patterns and mutagenesis data. We generated a homology model of Shigella flexneri needles compatible with both experimental data: the MxiH solid-state NMR chemical shifts and the state-of-the-art cryoEM density map. These results corroborate the solid-state NMR structure previously solved for Salmonella typhimurium PrgI needles and establish that Shigella flexneri and Salmonella typhimurium subunit proteins adopt a conserved structure and orientation in their

Escherichia coli and other Enterobacteriaceae: Food poisoning and health effects

Science.gov (United States)

The family Enterobactericeae consists of rod-shaped, Gram-negative, facultatively anaerobic, non-spore forming bacteria and also includes the food-borne pathogens, Cronobacter spp., Escherichia coli, Salmonella enterica, Shigella spp., and Yersinia spp. Illness caused by these pathogens is acquired...

Removal of salmonella-typhi, shigella-dysenteriae, vibrio-cholerae and rotavirus from water using a water-treatment tablet

CSIR Research Space (South Africa)

Rodda, N

1993-01-01

Full Text Available previously demonstrated. This study evaluated the efficiency of removal of Salmonella typhi, Shigella dysenteriae, Vibrio cholerae and rotavirus from simulated hard water of high organic content and colour. All four pathogenic micro organisms were...

Accurate differentiation of Escherichia coli and Shigella serogroups: challenges and strategies

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N.K. Devanga Ragupathi

2018-01-01

Full Text Available Shigella spp. and Escherichia coli are closely related; both belong to the family Enterobacteriaceae. Phenotypically, Shigella spp. and E. coli share many common characteristics, yet they have separate entities in epidemiology and clinical disease, which poses a diagnostic challenge. We collated information for the best possible approach to differentiate clinically relevant E. coli from Shigella spp. We found that a molecular approach is required for confirmation. High discriminatory potential is seen with whole genome sequencing analysed for k-mers and single nucleotide polymorphism. Among these, identification using single nucleotide polymorphism is easy to perform and analyse, and it thus appears more promising. Among the nonmolecular methods, matrix-assisted desorption ionization–time of flight mass spectrometry may be applicable when data analysis is assisted with advanced analytic tools. Keywords: 16S rRNA, k-mer, MALDI-TOF MS, single nucleotide polymorphism, whole genome sequencing

Virulence variations in Shigella and enteroinvasive Escherichia coli using the Caenorhabditis elegans model.

Science.gov (United States)

Fung, Crystal Ching; Octavia, Sophie; Mooney, Anne-Marie; Lan, Ruiting

2015-01-01

Shigella species and enteroinvasive Escherichia coli (EIEC) belong to the same species genetically, with remarkable phenotypic and genomic similarities. Shigella is the main cause of bacillary dysentery with around 160 million annual cases, while EIEC generally induces a milder disease compared to Shigella. This study aimed to determine virulence variations between Shigella and EIEC using the nematode Caenorhabditis elegans as a model host. Caenorhabditis elegans killing- and bacterial colonization assays were performed to examine the potential difference in virulence between Shigella and EIEC strains. Statistically significant difference in the survival rates of nematodes was demonstrated, with Shigella causing death at 88.24 ± 1.20% and EIEC at 94.37 ± 0.70%. The intestinal load of bacteria in the nematodes was found to be 7.65 × 10(4) ± 8.83 × 10(3) and 2.92 × 10(4) ± 6.26 × 10(3) CFU ml(-1) per nematode for Shigella and EIEC, respectively. Shigella dysenteriae serotype 1 which carries the Shiga toxin showed the lowest nematode survival rate at 82.6 ± 3.97% and highest bacterial colonization of 1.75 × 10(5) ± 8.17 × 10(4) CFU ml(-1), whereas a virulence plasmid-negative Shigella strain demonstrated 100 ± 0% nematode survival and lowest bacterial accumulation of 1.02 × 10(4) ± 7.23 × 10(2) CFU ml(-1). This study demonstrates C. elegans as an effective model for examining and comparing Shigella and EIEC virulence variation. © FEMS 2014. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.

[Susceptibility to azithromycin and other antibiotics in recent isolates of Salmonella, Shigella and Yersinia].

Science.gov (United States)

Martín-Pozo, Angeles; Arana, David M; Fuentes, Miriam; Alós, Juan-Ignacio

2014-01-01

Azithromycin represents an alternative option to treat bacterial diarrhea when the antibiotic therapy is indicated. Little is known regarding the susceptibility to azithromycin in enteropathogens in Spain. The MICs of azithromycin were determined by E-test against Salmonella non-typhi (SNT), Shigella and Yersinia isolates collected over the last three years (2010-2012). In addition, the susceptibility to other antibiotics usually used to treat gastrointestinal diseases was determined in these isolates by using a microdilution method. A total of 139 strains of SNT, Shigella and Yersinia were studied. All of them, except one strain, had a MIC≤16mg/L of azithromycin. In the adult population, 14.7% and 40.6% of SNT and Shigella isolates, respectively, were resistant to at least 2 of following antibiotics: amoxicillin, trimethoprim-sulfamethoxazole and ciprofloxacin. In the pediatric population, 10% of SNT clinical isolates and 28.6% (2/7) of Shigella isolates were resistant to amoxicillin and trimethoprim-sulfamethoxazole. In our experience, azithromycin would be a useful antibiotic alternative to treat bacterial diarrhea. Copyright © 2013 Elsevier España, S.L. y Sociedad Española de Enfermedades Infecciosas y Microbiología Clínica. All rights reserved.

Abortive Intestinal Infection With an Escherichia coli-Shigella flexneri Hybrid Strain

Science.gov (United States)

Formal, Samuel B.; LaBrec, E. H.; Kent, T. H.; Falkow, S.

1965-01-01

Formal, Samuel B., (Walter Reed Army Institute of Research, Washington, D.C.), E. H. LaBrec, T. H. Kent, and S. Falkow. Abortive intestinal infection with an Escherichia coli-Shigella flexneri hybrid strain. J. Bacteriol. 89:1374–1382. 1965.—The mechanism of the apparent loss of virulence of an Escherichia coli-Shigella flexneri hybrid strain was studied. The parent Shigella strain caused a fatal enteric infection when fed to starved guinea pigs, and signs of dysentery followed its oral administration to monkeys. The hybrid strain failed to produce any apparent symptoms when fed to either of these species. The parent strain was shown to invade the intestinal mucosa of starved guinea pigs. This caused a severe inflammatory reaction in the lamina propria, which progressed to ulceration of the intestinal epithelium and resulted in death of the animal. The hybrid strain also invaded the intestinal mucosa and produced an inflammatory reaction. In this case, the inflammatory reaction subsided, the intestine returned to normal within 4 days after challenge, and the animal survived. Both fluorescent-antibody techniques and in vivo growth studies have shown that the hybrid strain can not maintain itself in the intestinal mucosa. Preliminary studies have indicated that a similar situation also exists in the monkey. It is concluded that the virulence of dysentery bacilli rests not only in the capacity to reach the lamina propria, but also in the ability to multiply in this region. Images PMID:14293011

Shigella IpaH Family Effectors as a Versatile Model for Studying Pathogenic Bacteria.

Science.gov (United States)

Ashida, Hiroshi; Sasakawa, Chihiro

2015-01-01

Shigella spp. are highly adapted human pathogens that cause bacillary dysentery (shigellosis). Via the type III secretion system (T3SS), Shigella deliver a subset of virulence proteins (effectors) that are responsible for pathogenesis, with functions including pyroptosis, invasion of the epithelial cells, intracellular survival, and evasion of host immune responses. Intriguingly, T3SS effector activity and strategies are not unique to Shigella, but are shared by many other bacterial pathogens, including Salmonella, Yersinia, and enteropathogenic Escherichia coli (EPEC). Therefore, studying Shigella T3SS effectors will not only improve our understanding of bacterial infection systems, but also provide a molecular basis for developing live bacterial vaccines and antibacterial drugs. One of Shigella T3SS effectors, IpaH family proteins, which have E3 ubiquitin ligase activity and are widely conserved among other bacterial pathogens, are very relevant because they promote bacterial survival by triggering cell death and modulating the host immune responses. Here, we describe selected examples of Shigella pathogenesis, with particular emphasis on the roles of IpaH family effectors, which shed new light on bacterial survival strategies and provide clues about how to overcome bacterial infections.

Shigella IpaH family effectors as a versatile model for studying pathogenic bacteria

Directory of Open Access Journals (Sweden)

Hiroshi eAshida

2016-01-01

Full Text Available Shigella spp. are highly adapted human pathogens that cause bacillary dysentery (shigellosis. Via the type III secretion system (T3SS, Shigella deliver a subset of virulence proteins (effectors that are responsible for pathogenesis, with functions including pyroptosis, invasion of the epithelial cells, intracellular survival, and evasion of host immune responses. Intriguingly, T3SS effector activity and strategies are not unique to Shigella, but are shared by many other bacterial pathogens, including Salmonella, Yersinia, and enteropathogenic Escherichia coli (EPEC. Therefore, studying Shigella T3SS effectors will not only improve our understanding of bacterial infection systems, but also provide a molecular basis for developing live bacterial vaccines and antibacterial drugs. One of Shigella T3SS effectors, IpaH family proteins, which have E3 ubiquitin ligase activity and are widely conserved among other bacterial pathogens, are very relevant because they promote bacterial survival by triggering cell death and modulating the host immune responses. Here, we describe selected examples of Shigella pathogenesis, with particular emphasis on the roles of IpaH family effectors, which shed new light on bacterial survival strategies and provide clues about how to overcome bacterial infections.

Evaluation of antimicrobial resistance among Salmonella and Shigella isolates in the University Hospital "St. George," Plovdiv, Bulgaria.

Science.gov (United States)

Petrov, Michael M; Petrova, Atanaska; Stanimirova, Irina; Mircheva-Topalova, Marina; Koycheva, Lalka; Velcheva, Rayna; Stoycheva-Vartigova, Mariana; Raycheva, Ralitsa; Asseva, Galina; Petrov, Petar; Kardjeva, Velichka; Murdjeva, Marianna

2017-03-01

The aim of this work is to study the epidemiology and antimicrobial resistance to the most commonly used antibiotics for the treatment of acute gastroenteritis caused by Salmonella and Shigella at the largest Bulgarian hospital-University Hospital "St. George," Plovdiv-for the period 2009-2013. Two hundred ninety strains were in vitro tested for resistance to 15 antimicrobial agents. The presence of extended-spectrum beta-lactamases (ESBLs) was demonstrated by a variety of specialized tests. For comparison, a collection of 28 strains submitted by the National Reference Laboratory (NRL) "Enteric Infections" at the National Center of Infectious and Parasitic Diseases (NCIPD), Sofia, was also tested for the production of ESBLs. In isolates, phenotypically demonstrated as ESBL producers, polymerase chain reaction (PCR) detection of the genes bla-CTX-M, bla-SHV, and bla-TEM was performed. Among the 290 tested isolates, only two- Salmonella serotype Livingstone and Shigella flexneri-were phenotypically proven to be ESBL producers. Only 4 strains from the collection of 28, submitted from the NRL "Intestinal Infections" in NCIPD, Sofia, were phenotypically confirmed as ESBL producers. The presence of the bla-CTX-M gene was detected in all of the tested strains (4 from NRL, NCIPD, Sofia, and 2 from the University Hospital St. George, Plovdiv), the bla-SHV gene only in strain S. Livingstone from Plovdiv, and the bla-TEM gene in two from Sofia and one (again S. Livingstone) from Plovdiv. In conclusion, Salmonella and Shigella isolates from patients hospitalized at the University Hospital St. George, Plovdiv, with acute gastroenteritis demonstrate good susceptibility to the most commonly used antibiotic agents, including azithromycin.

Effects of subtherapeutic concentrations of antimicrobials on gene acquisition events in Yersinia, Proteus, Shigella, and Salmonella recipient organisms in isolated ligated intestinal loops of swine.

Science.gov (United States)

Brewer, Matt T; Xiong, Nalee; Anderson, Kristi L; Carlson, Steve A

2013-08-01

To assess antimicrobial resistance and transfer of virulence genes facilitated by subtherapeutic concentrations of antimicrobials in swine intestines. 20 anesthetized pigs experimentally inoculated with donor and recipient bacteria. 4 recipient pathogenic bacteria (Salmonella enterica serotype Typhimurium, Yersinia enterocolitica, Shigella flexneri, or Proteus mirabilis) were incubated with donor bacteria in the presence of subinhibitory concentrations of 1 of 16 antimicrobials in isolated ligated intestinal loops in swine. Donor Escherichia coli contained transferrable antimicrobial resistance or virulence genes. After coincubations, intestinal contents were removed and assessed for pathogens that acquired new antimicrobial resistance or virulence genes following exposure to the subtherapeutic concentrations of antimicrobials. 3 antimicrobials (apramycin, lincomycin, and neomycin) enhanced transfer of an antimicrobial resistance plasmid from commensal E coli organisms to Yersinia and Proteus organisms, whereas 7 antimicrobials (florfenicol, hygromycin, penicillin G, roxarsone, sulfamethazine, tetracycline, and tylosin) exacerbated transfer of an integron (Salmonella genomic island 1) from Salmonella organisms to Yersinia organisms. Sulfamethazine induced the transfer of Salmonella pathogenicity island 1 from pathogenic to nonpathogenic Salmonella organisms. Six antimicrobials (bacitracin, carbadox, erythromycin, sulfathiazole, tiamulin, and virginiamycin) did not mediate any transfer events. Sulfamethazine was the only antimicrobial implicated in 2 types of transfer events. 10 of 16 antimicrobials at subinhibitory or subtherapeutic concentrations augmented specific antimicrobial resistance or transfer of virulence genes into pathogenic bacteria in isolated intestinal loops in swine. Use of subtherapeutic antimicrobials in animal feed may be associated with unwanted collateral effects.

Investigating Shigella sonnei Infections

Centers for Disease Control (CDC) Podcasts

2011-11-17

Dr. Nancy Strockbine, Chief of the Escherichia and Shigella Reference Unit at CDC, discusses Shigella sonnei infections.  Created: 11/17/2011 by National Center for Emerging and Zoonotic Infectious Diseases (NCEZID).   Date Released: 11/21/2011.

Antibiotic resistant Salmonella and Escherichia coli isolated from ...

African Journals Online (AJOL)

Results: A hundred and four indigenous chicken rectal swabs were analysed, of which 67.3% were contaminated with Escherichia coli and 12.5% with Salmonella typhimurium. Seventy Escherichia coli isolates showed resistance phenotypes to one, two or more antibiotics. The most common antimicrobial resistance pattern ...

Vigilancia en red de los serotipos y la susceptibilidad antimicrobiana de Salmonella spp., Shigella spp. Y Vibrio cholerae O1, 1997 - 1999

Directory of Open Access Journals (Sweden)

Nélida Muñoz

2000-09-01

Full Text Available En 1997, el Grupo de Microbiología del INS estableció un programa en red con los Laboratorios de Salud Pública (LSP del país y el apoyo de la OPS, para la vigilancia de los principales patógenos causantes de enfermedad diarreica aguda. El objetivo fue conocer los serotipos y los patrones de resistencia antimicrobiana de Salmonella spp., Shigella spp. y Vibrio cholerae O1. Los aislamientos fueron confirmados de acuerdo con los esquemas de identificación bioquímica y serológica estandarizados y la determinación de la susceptibilidad antimicrobiana se realizó por la técnica de difusión de disco (Kirby-Bauer. De 1997 a 1999, participaron 22 LSP con el envio de 976 aislamientos, 96% de origen clínico y 4% de alimentos; 34% Salmonella spp., 23% Shigella spp. y 42% V. cholerae 01. La distribución por serotipo de Salmonella fue 39% S. Enteritidis, 27% S. Typhimurium, 9% grupo El, 5% S. Typhi y 20% otros serotipos; de los aislamientos de Shigella, 67% fueron S. grupo flexneriza, 30% S sonnei, 2% S. dysenteriae y 1% S. boydii. Para V. cholerae 01, 99% fue serotipo Ogawa. La susceptibilidad antimicrobiana determinó que 56% de los aislamientos de Salmonella eran resistentes y 22% multirresistentes, con un patrón predominante de ampicilina, tetraciclina y trimetoprim-sulfa (SXT. De los aislamientos de Shigella, 97% fueron resistentes y 57% multirresistentes, con un patrón de ampicilina, tetraciclina, cloranfenicol y SXT. No se observaron cambios en la susceptibilidad de V. cholerae O1. Este estudio enfatiza la importancia de continuar con el programa de vigilancia, para conocer la epidemiologia de la EDA en Colombia, darle un tratamiento óptimo a estas infecciones y poder diseñar programas para disminuir la diseminación de bacterias resistentes.

Serological cross-reaction between O-antigens of Shigella dysenteriae type 4 and an environmental Escherichia albertii isolate.

Science.gov (United States)

Rahman, Mohammed Ziaur; Akter, Selina; Azmuda, Nafisa; Sultana, Munawar; Weill, François-Xavier; Khan, Sirajul Islam; Grimont, Patrick A D; Birkeland, Nils-Kåre

2013-11-01

An environmental freshwater bacterial isolate, DM104, appearing as Shigella-like colonies on selective agar plates was found to show strong and specific serological cross-reactivity with Shigella dysenteriae type 4. Biochemical identification according to the analytical profile index, molecular serotyping by restriction of the amplified O-antigen gene cluster (rfb-RFLP), together with phylogenetic analysis of the 16S rRNA gene and multi-locus sequence analysis, identified the isolate as Escherichia albertii. rfb-RFLP of DM104, revealed a profile different from that of S. dysenteriae type 4. However, western blot analysis of extracted lipopolysaccharides demonstrated strong cross-reactivity with S. dysenteriae type 4 using specific monovalent antisera and a lipopolysaccharide gel banding profile similar to that of S. dysenteriae type 4. The observed O-antigen cross-reaction between an E. albertii isolate and S. dysenteriae extends our knowledge of the extent of O-antigen cross-reaction within the Escherichia/Shigella group of organisms, and offers the possibility of using DM104 and similar cross-reacting strains as shigellosis vaccine candidates.

Nota sobre infecção por Salmonella sp. e Shigella sp. em bugios-ruivos, Alouatta clamitans Cabrera, 1940 (Primates: Atelidae mantidos em cativeiro

Directory of Open Access Journals (Sweden)

Júlio César de Souza Júnior

2008-08-01

Full Text Available Amostras de fezes de bugios-ruivos cativos foram investigadas quanto a presença de Salmonella sp. e Shigella sp. O padrão de sensibilidade a antibióticos foi testado. Este é o primeiro relato de infecção assintomática por Salmonella sp. nesta espécie. Não foi detectada infecção por Shigella sp. Destaca-se a importância de investigação em animais submetidos a translocação ou a reintrodução e a necessidade de inquéritos periódicos em populações cativas a fim de se evitar agravos ocupacionais.

Simultaneous and individual quantitative estimation of Salmonella, Shigella and Listeria monocytogenes on inoculated Roma tomatoes (Lycopersicon esculentum var. Pyriforme) and Serrano peppers (Capsicum annuum) using an MPN technique.

Science.gov (United States)

Cabrera-Díaz, E; Martínez-Chávez, L; Sánchez-Camarena, J; Muñiz-Flores, J A; Castillo, A; Gutiérrez-González, P; Arvizu-Medrano, S M; González-Aguilar, D G; Martínez-Gonzáles, N E

2018-08-01

Simultaneous and individual enumeration of Salmonella, Shigella and Listeria monocytogenes was compared on inoculated Roma tomatoes and Serrano peppers using an Most Probable Number (MPN) technique. Samples consisting of tomatoes (4 units) or peppers (8 units) were individually inoculated with a cocktail of three strains of Salmonella, Shigella or L. monocytogenes, or by simultaneous inoculation of three strains of each pathogen, at low (1.2-1.7 log CFU/sample) and high (2.2-2.7 log CFU/sample) inocula. Samples were analyzed by an MPN technique using universal pre-enrichment (UP) broth at 35 °C for 24 ± 2 h. The UP tubes from each MPN series were transferred to enrichment and plating media following adequate conventional methods for isolating each pathogen. Data were analyzed using multifactorial analysis of variance (p  simultaneous), type of bacteria (Salmonella > Shigella and L. monocytogenes), type of sample (UP broth > pepper and tomato), and inoculum level (high > low). The MPN technique was effective for Salmonella on both commodities. Shigella counts were higher on tomatoes compared to peppers, (p < 0.05), and for L. monocytogenes on peppers (p < 0.05). Copyright © 2018 Elsevier Ltd. All rights reserved.

Whole-genome phylogeny of Escherichia coli/Shigella group by feature frequency profiles (FFPs)

Science.gov (United States)

Sims, Gregory E.; Kim, Sung-Hou

2011-01-01

A whole-genome phylogeny of the Escherichia coli/Shigella group was constructed by using the feature frequency profile (FFP) method. This alignment-free approach uses the frequencies of l-mer features of whole genomes to infer phylogenic distances. We present two phylogenies that accentuate different aspects of E. coli/Shigella genomic evolution: (i) one based on the compositions of all possible features of length l = 24 (∼8.4 million features), which are likely to reveal the phenetic grouping and relationship among the organisms and (ii) the other based on the compositions of core features with low frequency and low variability (∼0.56 million features), which account for ∼69% of all commonly shared features among 38 taxa examined and are likely to have genome-wide lineal evolutionary signal. Shigella appears as a single clade when all possible features are used without filtering of noncore features. However, results using core features show that Shigella consists of at least two distantly related subclades, implying that the subclades evolved into a single clade because of a high degree of convergence influenced by mobile genetic elements and niche adaptation. In both FFP trees, the basal group of the E. coli/Shigella phylogeny is the B2 phylogroup, which contains primarily uropathogenic strains, suggesting that the E. coli/Shigella ancestor was likely a facultative or opportunistic pathogen. The extant commensal strains diverged relatively late and appear to be the result of reductive evolution of genomes. We also identify clade distinguishing features and their associated genomic regions within each phylogroup. Such features may provide useful information for understanding evolution of the groups and for quick diagnostic identification of each phylogroup. PMID:21536867

Prevalence of Stx-producing Shigella species isolated from French Travelers Returning from the Caribbean: An Emerging Pathogen with International Implications

Science.gov (United States)

Gray, Miranda D.; Lacher, David W.; Leonard, Susan R.; Abbott, Jason; Zhao, Shaohua; Lampel, Keith A.; Prothery, Estelle; Gouali, Malika; Weill, François-Xavier; Maurelli, Anthony T.

2015-01-01

Shiga toxins are potent cytotoxins that inhibit host cell protein synthesis, leading to cell death. Classically, these toxins are associated with intestinal infections due to Shiga toxin-producing Escherichia coli or Shigella dysenteriae serotype 1 and infections with these strains can lead to hemolytic uremic syndrome. Over the past decade there is increasing recognition that Shiga toxin is produced by additional Shigella species. We recently reported the presence and expression of stx genes in Shigella flexneri 2a clinical isolates. The toxin genes were carried by a new stx-encoding bacteriophage and infection with these strains correlated with recent travel to Haiti or the Dominican Republic. In this study we further explored the epidemiological link to this region by utilizing the French National Reference Center for Escherichia coli, Shigella and Salmonella collection to survey the frequency of Stx-producing Shigella species isolated from French travelers returning from the Caribbean. About 21% of the isolates tested were found to encode and produce Stx. These isolates included strains of S. flexneri 2a, S. flexneri Y, and S. dysenteriae 4. All of the travelers whom were infected with Stx-producing Shigella had recently traveled to Haiti, the Dominican Republic, or French Guiana. Furthermore, whole genome sequencing found that the toxin genes were encoded by a prophage that was highly identical to the phage we identified in our previous study. These findings demonstrate that this new stx-encoding prophage is circulating within that geographical area, has spread to other continents, and is capable of spreading to multiple Shigella serogroups. PMID:25980352

Detection of Salmonella Spp., Shigella (Flexneri and Sonnei) and Vibrio Cholerae O1 by Polymerase Chain Reaction (PCR) in Exported Shrimp from the Mexican Northeast Coast

Energy Technology Data Exchange (ETDEWEB)

Perez, L.; Nuñez, F.; Rubio, M.; Nicoli, M. [Universidad Nacional Autónoma de México, Facultad de Medicina Veterinaria y Zootecnia (Mexico)

2005-01-15

The objective of the present work was to use the PCR technique for the simultaneous detection of Salmonella spp and Vibrio cholerae O1 in frozen shrimp for export. The DNA segments located in the gene A [284 pairs of bases (pb)] from Salmonella spp. locus ial (217 and 320 pb) from Shigella flexneri and Shigella sonnei and the gene ctxA and ctxB (777 pb) from Vibrio cholerae O1 were amplified. The different primers that amplify these segments were assayed in a PCR reaction for the simultaneous detection of DNA from the microorganisms. It was not possible to amplify the gene of Shigella sonnei and Shigella flexneri under the assay’s conditions, whilst those of Salmonella spp. and Vibrio cholerae O1 were successfully amplified. The amplification conditions for the PCR were: 94° C, 58° C and 72° C during 30 cycles, allowing a reduction from 15 days test time with the official microbiological methods to 28 hours (24 for the pre-enrichment and four for the PCR). Samples of raw-frozen-headless shrimps were taken from production plants located in the State of Sinaloa, Mexico. A random sampling procedure was used, according to the guidelines described by the International Commission of Microbiological Specifications for Foods (ICMSF, 1999). Five packages per lot per production plant were obtained. From each individual package (5 pounds 80 OZ ≈ 2.27 kg) three samples were taken for the bacteriological assays to search for Salmonella spp. and Vibrio cholerae O1, respectively. The samples were also analyzed by PCR. Results showed that none of the samples were positive by PCR to any of the studied bacteria. Salmonella spp. and Vibrio cholerae O1 were not detected in these samples by the official methods. However, the latter were able to identify other Vibrio species and enterobacteria like Proteus and Acromobacter. These results confirmed PCR’s rapidity, sensitivity and specificity. (author)

Inhibitory Activity of Lactid Acid Bacteria Isolated from Tape Waterlily Seed to Enteric Pathogenic Bacteria (Vibrio cholera, Salmonella typhi, Shigella disentri, and E.coli and Its’ Susceptibility to Antibiotic, Bile Salt and Acidic Condition

Directory of Open Access Journals (Sweden)

Iin Khusnul Khotimah

2012-03-01

Full Text Available The aim of this research was to observe inhibitory activity of LAB isolated from tape waterlily seed to enteric pathogenic bacteria (Vibrio cholera, Salmonella typhi, Shigella disentri, E.coli ATCC 25922 and it’s susceptibility to antibiotic, in bile salt and under acidic condition. Microbia in the tape ( a fermented product of waterlily seed to showed were Streptococcus thermophilus (IKH-1, Pediococcus pentosaceus (IKH-2 and Leuconostoc mesentroides (IKH-8. Streptococcus thermophillus showed inhibition against the growth of Shigella disentri with inhibition zones 16,28 mm, but did not against the growth of V. Cholera, S. typhi, E.coli. Pediococcus pentosaceus inhibit Vibrio cholera, dan Salmonella thypi with inhibition zones 18,59 mm dan 7,91 mm. So that, Leuconostoc mesenteroides inhibit Salmonella thypi with zones inhibits average 8,25 mm. Chloramfenicol at 0.05 mg concentrations did not show inhibition against the growth of isolated Streptococcus thermophillus, Pediococcus pentosaceus and Leuconostoc mesentroides. These isolates could survive too in bile salt (2% and acidified media (pH 3.   Keyword : The tape of  waterlily seed, LAB, probiotic and enteric pathogenic   KEMAMPUAN PENGHAMBATAN BAKTERI ASAM LAKTAT DARI TAPE BIJI TERATAI TERHADAP PATOGENIK ENTERIK (VIBRIO CHOLERA, SALMONELLA THYPI, SHIGELLA DISENTRI, E. COLI, ANTIBIOTIK, KETAHANANNYA TERHADAP BILE SALT DAN ASAM   ABSTRAK   Penelitian ini bertujuan untuk menguji kemampuan penghambatan bakteri asam laktat yang diisolasi dari tape biji teratai terhadap patogenik enterik (Vibrio cholera, Salmonella thypi, Shigella disentri, E. Coli ATCC 25922, antibiotik, bile salt dan asam. Jenis bakteri yang diketahui tumbuh selama fermentasi tape biji teratai adalah Streptococcus thermopilus (IKH-1, Pediococcus pentosaceus(IKH-2, dan Leuconostoc mesentroides (IKH-8. Pengamatan terhadap uji penghambatan patogenik enterik (Vibrio cholera, Salmonella thypi, Shigella disentri, dan E. Coli ATCC

Salmonella serotypeTyphi, Shigella, and intestinal parasites among food handlers at Bahir Dar University, Ethiopia.

Science.gov (United States)

Abera, Bayeh; Yitayew, Gashaw; Amare, Hiwot

2016-02-28

Food handlers play a major role in the transmission of Salmonella serotype Typhi (S. Typhi), Shigella, and intestinal parasites. This study was conducted to determine the prevalence of S. Typhi, Shigella, and intestinal parasites among food handlers at Bahir Dar University, Ethiopia. A cross-sectional study was conducted in June 2014. Stool samples from 410 food handlers were examined for bacterial pathogens and parasites. Pearson's Chi-square test, Fisher's exact test, and bivariate and multivariate logistic regression analyses were used where appropriate. The prevalence of S. Typhi, Shigella, and intestinal parasites among food handlers was 11 (2.7%), 5 (1.2%), and 53 (12.9%), respectively. Among eight intestinal parasites identified, the two most prevalent intestinal parasites were hookworm 26 (6.3%) and G. lamblia 13 (3.1%). Male food handlers were more likely to be positive than were female food handlers for S. Typhi and intestinal parasites. Furthermore, food handlers who had a history of regular medical checkups were less infected with intestinal parasites. Being male (AOR: 2.1, 95% CI: 1.2, 4.4) and not attending medical checkups (AOR: 2.9, 95% CI: 1.4, 6.1) were independent predictors of intestinal parasitic infection in food handlers. Male food handlers were reluctant to have regular parasitological examinations. There was a high proportion of food handlers with S. Typhi, Shigella, and intestinal parasites in their faces. Special emphasis should be placed on S. Typhicarriers and male food handlers. Education and periodical medical checkups for intestinal parasites and S. Typhi should be considered as intervention measures.

Inhibition of Escherichia Coli, Salmonella and Staphylococcus ...

African Journals Online (AJOL)

Escherichia coli O157:H7, Salmonella typhimurium and Staphylococcus. aureus are of great concern to the food industry, especially in foods stored under refrigerated conditions where, unlike most food-borne pathogens are able to multiply. This investigation was conducted to study the inhibitory effect of some spice ...

Multicenter evaluation of molecular and culture-dependent diagnostics for Shigella species and Entero-invasive Escherichia coli in the Netherlands

NARCIS (Netherlands)

van den Beld, Maaike J. C.; Friedrich, Alexander W.; van Zanten, Evert; Reubsaet, Frans A. G.; Kooistra-Smid, Mirjam A. M. D.; Rossen, John W. A.

2016-01-01

An inter-laboratory collaborative trial for the evaluation of diagnostics for detection and identification of Shigella species and Entero-invasive Escherichia coil (EIEC) was performed. Sixteen Medical Microbiological Laboratories (MMLs) participated. MMLs were interviewed about their diagnostic

Comparative sequence analysis of acid sensitive/resistance proteins in Escherichia coli and Shigella flexneri

Science.gov (United States)

Manikandan, Selvaraj; Balaji, Seetharaaman; Kumar, Anil; Kumar, Rita

2007-01-01

The molecular basis for the survival of bacteria under extreme conditions in which growth is inhibited is a question of great current interest. A preliminary study was carried out to determine residue pattern conservation among the antiporters of enteric bacteria, responsible for extreme acid sensitivity especially in Escherichia coli and Shigella flexneri. Here we found the molecular evidence that proved the relationship between E. coli and S. flexneri. Multiple sequence alignment of the gadC coded acid sensitive antiporter showed many conserved residue patterns at regular intervals at the N-terminal region. It was observed that as the alignment approaches towards the C-terminal, the number of conserved residues decreases, indicating that the N-terminal region of this protein has much active role when compared to the carboxyl terminal. The motif, FHLVFFLLLGG, is well conserved within the entire gadC coded protein at the amino terminal. The motif is also partially conserved among other antiporters (which are not coded by gadC) but involved in acid sensitive/resistance mechanism. Phylogenetic cluster analysis proves the relationship of Escherichia coli and Shigella flexneri. The gadC coded proteins are converged as a clade and diverged from other antiporters belongs to the amino acid-polyamine-organocation (APC) superfamily. PMID:21670792

Establishing Streptomycin Epidemiological Cut-Off Values for Salmonella and Escherichia coli

DEFF Research Database (Denmark)

Migura, Lourdes Garcia; Sunde, Marianne; Karlsmose, Susanne

2011-01-01

This study was conducted to elucidate the accuracy of the current streptomycin epidemiological cut-off value (ECOFF) for Escherichia coli and Salmonella spp. A total of 236 Salmonella enterica and 208 E. coli isolates exhibiting MICs between 4 and 32 mg/L were selected from 12 countries. Isolates...

Relationship among fecal coliforms, Escherichia coli, and Salmonella spp. in shellfish.

OpenAIRE

Hood, M A; Ness, G E; Blake, N J

1983-01-01

The relationship of fecal coliforms, Escherichia coli, and Salmonella spp. was examined in freshly harvested and stored shellfish. In 16 of 40 freshly collected oyster samples, fecal coliform levels were above the recommended wholesale level suggested by the National Shellfish Sanitation Program (less than or equal to 230/100 g), and Salmonella spp. were present in three of these samples. Salmonella spp. were not, however, present in any sample containing less than 230 fecal coliforms per 100...

Multicenter evaluation of molecular and culture-dependent diagnostics for Shigella species and Entero-invasive Escherichia coli in the Netherlands.

Science.gov (United States)

van den Beld, Maaike J C; Friedrich, Alexander W; van Zanten, Evert; Reubsaet, Frans A G; Kooistra-Smid, Mirjam A M D; Rossen, John W A

2016-12-01

An inter-laboratory collaborative trial for the evaluation of diagnostics for detection and identification of Shigella species and Entero-invasive Escherichia coli (EIEC) was performed. Sixteen Medical Microbiological Laboratories (MMLs) participated. MMLs were interviewed about their diagnostic methods and a sample panel, consisting of DNA-extracts and spiked stool samples with different concentrations of Shigella flexneri, was provided to each MML. The results of the trial showed an enormous variety in culture-dependent and molecular diagnostic techniques currently used among MMLs. Despite the various molecular procedures, 15 out of 16 MMLs were able to detect Shigella species or EIEC in all the samples provided, showing that the diversity of methods has no effect on the qualitative detection of Shigella flexneri. In contrast to semi quantitative analysis, the minimum and maximum values per sample differed by approximately five threshold cycles (Ct-value) between the MMLs included in the study. This indicates that defining a uniform Ct-value cut-off for notification to health authorities is not advisable. Copyright © 2016 Elsevier B.V. All rights reserved.

Evolutionary Dynamics of Small RNAs in 27 Escherichia coli and Shigella Genomes

Science.gov (United States)

Skippington, Elizabeth; Ragan, Mark A.

2012-01-01

Small RNAs (sRNAs) are widespread in bacteria and play critical roles in regulating physiological processes. They are best characterized in Escherichia coli K-12 MG1655, where 83 sRNAs constitute nearly 2% of the gene complement. Most sRNAs act by base pairing with a target mRNA, modulating its translation and/or stability; many of these RNAs share only limited complementarity to their mRNA target, and require the chaperone Hfq to facilitate base pairing. Little is known about the evolutionary dynamics of bacterial sRNAs. Here, we apply phylogenetic and network analyses to investigate the evolutionary processes and principles that govern sRNA gene distribution in 27 E. coli and Shigella genomes. We identify core (encoded in all 27 genomes) and variable sRNAs; more than two-thirds of the E. coli K-12 MG1655 sRNAs are core, whereas the others show patterns of presence and absence that are principally due to genetic loss, not duplication or lateral genetic transfer. We present evidence that variable sRNAs are less tightly integrated into cellular genetic regulatory networks than are the core sRNAs, and that Hfq facilitates posttranscriptional cross talk between the E. coli–Shigella core and variable genomes. Finally, we present evidence that more than 80% of genes targeted by Hfq-associated core sRNAs have been transferred within the E. coli–Shigella clade, and that most of these genes have been transferred intact. These results suggest that Hfq and sRNAs help integrate laterally acquired genes into established regulatory networks. PMID:22223756

In vitro antibacterial activity of methanol and water extracts of adiantum capillus veneris and tagetes patula against multidrug resistant bacterial strains

International Nuclear Information System (INIS)

Hussain, M.M.; Ahmad, B.; Bashid, E.; Hashim, S.

2014-01-01

The aim of present study was to screen the antimicrobial activities of extracts of leaves and stems of Adiantum capillus veneris and Tagetes patula against multidrug-resistant (MDR) bacterial strains. Extracts from the leaves and stems of these plants were extracted with methanol and water and tested for their antibacterial activity by disc diffusion method against ten MDR bacterial strains i.e., Citrobacter freundii, Escherichia coli, Providencia, Pseudomonas aeruginosa, Staphylococcus aureus, Klebsiella pneumoniae, Proteus vulgaris, Salmonella typhi, Shigella and Vibrio cholerae. Leaves methanol extract (LME) of Adiantum showed maximum Zone of Inhibition (ZI) against Providencia, Klebsiella pneumoniae, Shigella, Vibrio cholerae, Staphylococcus aureus, Proteus vulgaris and Salmonella typhi, whereas its stem methanol extract (SME) was very active against Escherichia coli, Klebsiella pneumoniae and Salmonella typhi. Similarly LME of Tagetes showed highest ZI against Escherichia coli and Vibrio cholerae while SME showed highest ZI to Escherichia coli, Vibrio cholerae, Providencia, Shigella and Klebsiella pneumoniae. Leaves water extract (LWE) of Adiantum was very active against all ten bacterial strains while its stem water extract (SWE) showed maximum ZI against Escherichia coli, Klebsiella pneumoniae and Salmonella typhi, Shigella, Proteus vulgaris and Providencia. LWE of Tagetes was only active against Vibrio cholerae whereas SWE was very active against Salmonella typhi and active against P. vulgaris, Citrobacter freundii and Vibrio cholerae. It was concluded from this study that extracts of both Adiantum and Tagetes have prominent activities against most of the MDR bacterial strains and needs further studies for utmost benefits. (author)

Investigating the Relatedness of Enteroinvasive Escherichia coli to Other E. coli and Shigella Isolates by Using Comparative Genomics.

Science.gov (United States)

Hazen, Tracy H; Leonard, Susan R; Lampel, Keith A; Lacher, David W; Maurelli, Anthony T; Rasko, David A

2016-08-01

Enteroinvasive Escherichia coli (EIEC) is a unique pathovar that has a pathogenic mechanism nearly indistinguishable from that of Shigella species. In contrast to isolates of the four Shigella species, which are widespread and can be frequent causes of human illness, EIEC causes far fewer reported illnesses each year. In this study, we analyzed the genome sequences of 20 EIEC isolates, including 14 first described in this study. Phylogenomic analysis of the EIEC genomes demonstrated that 17 of the isolates are present in three distinct lineages that contained only EIEC genomes, compared to reference genomes from each of the E. coli pathovars and Shigella species. Comparative genomic analysis identified genes that were unique to each of the three identified EIEC lineages. While many of the EIEC lineage-specific genes have unknown functions, those with predicted functions included a colicin and putative proteins involved in transcriptional regulation or carbohydrate metabolism. In silico detection of the Shigella virulence plasmid (pINV), which is essential for the invasion of host cells, demonstrated that a form of pINV was present in nearly all EIEC genomes, but the Mxi-Spa-Ipa region of the plasmid that encodes the invasion-associated proteins was absent from several of the EIEC isolates. The comparative genomic findings in this study support the hypothesis that multiple EIEC lineages have evolved independently from multiple distinct lineages of E. coli via the acquisition of the Shigella virulence plasmid and, in some cases, the Shigella pathogenicity islands. Copyright © 2016, American Society for Microbiology. All Rights Reserved.

Multiplex Polymerase Chain Reaction for Identification of Shigellae and Four Shigella Species Using Novel Genetic Markers Screened by Comparative Genomics.

Science.gov (United States)

Kim, Hyun-Joong; Ryu, Ji-Oh; Song, Ji-Yeon; Kim, Hae-Yeong

2017-07-01

In the detection of Shigella species using molecular biological methods, previously known genetic markers for Shigella species were not sufficient to discriminate between Shigella species and diarrheagenic Escherichia coli. The purposes of this study were to screen for genetic markers of the Shigella genus and four Shigella species through comparative genomics and develop a multiplex polymerase chain reaction (PCR) for the detection of shigellae and Shigella species. A total of seven genomic DNA sequences from Shigella species were subjected to comparative genomics for the screening of genetic markers of shigellae and each Shigella species. The primer sets were designed from the screened genetic markers and evaluated using PCR with genomic DNAs from Shigella and other bacterial strains in Enterobacteriaceae. A novel Shigella quintuplex PCR, designed for the detection of Shigella genus, S. dysenteriae, S. boydii, S. flexneri, and S. sonnei, was developed from the evaluated primer sets, and its performance was demonstrated with specifically amplified results from each Shigella species. This Shigella multiplex PCR is the first to be reported with novel genetic markers developed through comparative genomics and may be a useful tool for the accurate detection of the Shigella genus and species from closely related bacteria in clinical microbiology and food safety.

Development of a Threat Assessment Framework Applicable to Dual Use Biotechnology: Results of a Study to Determine the Feasibility, Applicability and Potential Design of a Threat Assessment Framework Concept

Science.gov (United States)

2007-04-01

Escherichia coli, pathogenic Vibrio spp., Shigella spp., Salmonella spp., Listeria monocytogenes, Campylobacter jejuni and Yersinia enterocolitica...Marburg virus V12. Monkey pox virus V13. Rift Valley fever virus V14. Tick-borne...Burkholderia pseudomallei (Pseudomonas pseudomallei) B10. Salmonella typhi B11. Shigella dysenteriae B12

Vaccines for viral and bacterial pathogens causing acute gastroenteritis: Part II: Vaccines for Shigella, Salmonella, enterotoxigenic E. coli (ETEC) enterohemorragic E. coli (EHEC) and Campylobacter jejuni

Science.gov (United States)

O’Ryan, Miguel; Vidal, Roberto; del Canto, Felipe; Carlos Salazar, Juan; Montero, David

2015-01-01

In Part II we discuss the following bacterial pathogens: Shigella, Salmonella (non-typhoidal), diarrheogenic E. coli (enterotoxigenic and enterohemorragic) and Campylobacter jejuni. In contrast to the enteric viruses and Vibrio cholerae discussed in Part I of this series, for the bacterial pathogens described here there is only one licensed vaccine, developed primarily for Vibrio cholerae and which provides moderate protection against enterotoxigenic E. coli (ETEC) (Dukoral®), as well as a few additional candidates in advanced stages of development for ETEC and one candidate for Shigella spp. Numerous vaccine candidates in earlier stages of development are discussed. PMID:25715096

Relationship among Shigella spp. and enteroinvasive Escherichia coli (EIEC) and their differentiation.

Science.gov (United States)

Ud-Din, Abu; Wahid, Syeda

2014-01-01

Shigellosis produces inflammatory reactions and ulceration on the intestinal epithelium followed by bloody or mucoid diarrhea. It is caused by enteroinvasive E. coli (EIEC) as well as any species of the genus Shigella, namely, S. dysenteriae, S. flexneri, S. boydii, and S. sonnei. This current species designation of Shigella does not specify genetic similarity. Shigella spp. could be easily differentiated from E. coli, but difficulties observed for the EIEC-Shigella differentiation as both show similar biochemical traits and can cause dysentery using the same mode of invasion. Sequencing of multiple housekeeping genes indicates that Shigella has derived on several different occasions via acquisition of the transferable forms of ancestral virulence plasmids within commensal E. coli and form a Shigella-EIEC pathovar. EIEC showed lower expression of virulence genes compared to Shigella, hence EIEC produce less severe disease than Shigella spp. Conventional microbiological techniques often lead to confusing results concerning the discrimination between EIEC and Shigella spp. The lactose permease gene (lacY) is present in all E. coli strains but absent in Shigella spp., whereas β-glucuronidase gene (uidA) is present in both E. coli and Shigella spp. Thus uidA gene and lacY gene based duplex real-time PCR assay could be used for easy identification and differentiation of Shigella spp. from E. coli and in particular EIEC.

Molecular Mechanisms of Host Cytoskeletal Rearrangements by Shigella Invasins

Directory of Open Access Journals (Sweden)

Jun Hyuck Lee

2014-10-01

Full Text Available Pathogen-induced reorganization of the host cell cytoskeleton is a common strategy utilized in host cell invasion by many facultative intracellular bacteria, such as Shigella, Listeria, enteroinvasive E. coli and Salmonella. Shigella is an enteroinvasive intracellular pathogen that preferentially infects human epithelial cells and causes bacillary dysentery. Invasion of Shigella into intestinal epithelial cells requires extensive remodeling of the actin cytoskeleton with the aid of pathogenic effector proteins injected into the host cell by the activity of the type III secretion system. These so-called Shigella invasins, including IpaA, IpaC, IpgB1, IpgB2 and IpgD, modulate the actin-regulatory system in a concerted manner to guarantee efficient entry of the bacteria into host cells.

An assessment of enterotoxigenic Escherichia coli and Shigella vaccine candidates for infants and children.

Science.gov (United States)

Walker, Richard I

2015-02-18

Despite improvements to water quality, sanitation, and the implementation of current prevention and treatment interventions, diarrhea remains a major cause of illness and death, especially among children less than five years of age in the developing world. Rotavirus vaccines have already begun making a real impact on diarrhea, but several more enteric vaccines will be necessary to achieve broader reductions of illness and death. Among the many causes of diarrheal disease, enterotoxigenic Escherichia coli (ETEC) and Shigella are the two most important bacterial pathogens for which there are no currently licensed vaccines. Vaccines against these two pathogens could greatly reduce the impact of disease caused by these infections. This review describes the approaches to ETEC and Shigella vaccines that are currently under development, including a range of both cellular and subunit approaches for each pathogen. In addition, the review discusses strategies for maximizing the potential benefit of these vaccines, which includes the feasibility of co-administration, consolidation, and combination of vaccine candidates, as well as issues related to effective administration of enteric vaccines to infants. Recent impact studies indicate that ETEC and Shigella vaccines could significantly benefit global public health. Either vaccine, particularly if they could be combined together or with another enteric vaccine, would be an extremely valuable tool for saving lives and promoting the health of infants and children in the developing world, as well as potentially providing protection to travelers and military personnel visiting endemic areas. Copyright © 2014 Elsevier Ltd. All rights reserved.

Isolation and Antimicrobial Susceptibility Profile of Shigella and ...

African Journals Online (AJOL)

2018-03-01

Mar 1, 2018 ... species, Vibrio cholera, Yersinia enterocolitica, and Aeromonas species), enteroparasites (Giardia lamblia, Cryptosporidium species and Entamoeba histolytica), and viruses (adenovirus, Norwalk virus, and rotavirus) (5). Among the bacterial causative agents,. Salmonella and Shigella remain the major.

Identification and antigenic characterization of virulence-associated, plasmid-coded proteins of Shigella spp. and enteroinvasive Escherichia coli.

OpenAIRE

Hale, T L; Oaks, E V; Formal, S B

1985-01-01

Seven plasmid-coded polypeptides, designated a through g, were identified by two-dimensional nonequilibrium pH gradient electrophoresis of radiolabeled extracts from minicells of virulent Shigella flexneri serotypes 2a and 5 and enteroinvasive Escherichia coli O143. These polypeptides were deemed to be products of 140-megadalton (MDa) virulence-associated plasmids because they were not synthesized in minicells which were not harboring a 140-MDa plasmid or in minicells which were carrying an F...

Polymerase chain reaction for detection of invasive Shigella flexneri in food.

OpenAIRE

Lampel, K A; Jagow, J A; Trucksess, M; Hill, W E

1990-01-01

The polymerase chain reaction (PCR) was used to amplify a 760-base-pair (bp) fragment with the 220-kbp invasive plasmids of enteroinvasive Escherichia coli, Shigella flexneri, Shigella dysenteriae, Shigella boydii, and Shigella sonnei as templates. This PCR product was easily detected by agarose gel electrophoresis. A 210-bp AccI-PstI fragment lying within the amplified region was used as a probe in Southern hybridization blots and showed that the PCR-generated product was derived from the in...

Behavior of Salmonella Typhimurium, Staphylococcus aureus, Listeria monocytogenes, and Shigella flexneri and Shigella sonnei during production of pulque, a traditional Mexican beverage.

Science.gov (United States)

Gómez-Aldapa, Carlos A; Díaz-Cruz, Claudio A; Villarruel-López, Angélica; Torres-Vitela, M del Refugio; Añorve-Morga, Javier; Rangel-Vargas, Esmeralda; Cerna-Cortes, Jorge F; Vigueras-Ramírez, J Gabriel; Castro-Rosas, Javier

2011-04-01

Pulque is a typical fermented alcoholic beverage of central Mexico, produced from the nectar of maguey agave plants. Production systems are largely artisanal, with inadequate hygiene conditions and exposure to multiple contamination sources. No data exist on pulque microbiological safety and the behavior of pathogenic microorganisms in agave nectar and pulque. An initial trial was done of the behavior of Salmonella Typhimurium, Staphylococcus aureus, Listeria monocytogenes, and Shigella flexneri and Shigella sonnei during fermentation of nectar from a single producer, nectar mixture from different producers, and seed pulque. A second trial simulating artisanal pulque production was done by contaminating fresh nectar with each of the five strains, storing at 22°C for 14 h, adding seed pulque, and fermenting until pulque was formed. During incubation at 16 or 22°C in the first trial, all the pathogenic strains multiplied in both the single producer nectar and the nectar mixture, reaching maximum concentrations at 12 h. Strains concentration then decreased slowly. In the seed pulque, the strains did not multiply and tended to die. In the second trial, all strains increased concentration from 0.7 to 1.6 log at 22°C, and from 0.5 to 1.1 at 16°C in the first 14 h. After addition of seed pulque, they were quickly deactivated until none was detected in the final product. The results suggest that the potential risk to consumers of contracting any of the five tested pathogenic bacterial strains from pulque is low.

Relationship among fecal coliforms, Escherichia coli, and Salmonella spp. in shellfish.

Science.gov (United States)

Hood, M A; Ness, G E; Blake, N J

1983-01-01

The relationship of fecal coliforms, Escherichia coli, and Salmonella spp. was examined in freshly harvested and stored shellfish. In 16 of 40 freshly collected oyster samples, fecal coliform levels were above the recommended wholesale level suggested by the National Shellfish Sanitation Program (less than or equal to 230/100 g), and Salmonella spp. were present in three of these samples. Salmonella spp. were not, however, present in any sample containing less than 230 fecal coliforms per 100 g. Analysis of the data suggests that low fecal coliform levels in both fresh and stored oysters are good indicators of the absence of Salmonella spp., but that high levels of fecal coliforms are somewhat limited in predicting the presence of Salmonella spp. E. coli levels correlated very strongly with fecal coliform levels in both fresh and stored oysters and clams, suggesting that there is no advantage in replacing fecal coliforms with E. coli as an indicator of shellfish quality.

Application of DNA hybridization techniques in the assessment of diarrheal disease among refugess in Thailand. [Shigella; Escherichia coli; Campylobacter; Cryptosporidium

Energy Technology Data Exchange (ETDEWEB)

Taylor, D.N.; Echeverria, P.; Pitarangsi, C.; Seriwatana, J.; Sethabutr, O.; Bodhidatta, L.; Brown, C.; Herrmann, J.E.; Blacklow, N.R.

1988-01-01

The epidemiology and etiology of acute diarrheal disease were determined in a Hmong refugee camp on the Thai-Laotian border from April 11 to May 14, 1985. DNA hybridization techniques were used to detect Shigella species, enteroinvasive Escherichia coli, and enterotoxigenic E. coli. A monoclonal enzyme-linked immunosorbent assay was used to detect rotavirus, and standard microbiology was used to detect other enteropathogens. The age-specific diarrheal disease rates were 47 episodes per month per 1000 children less than five years old and 113 episodes per month per 1000 children less than one year old. Rotavirus, enterotoxigenic E. coli, Campylobacter, and Cryptosporidium were the predominant pathogens in children less than two years old. The DNA probe hybridized with 94% of 31 specimens identified as enterotoxigenic E. coli by the standard assays and with none of the specimens in which the standard assays were negative. The probe for Shigella and enteroinvasive E. coli hybridized in eight of 10 stools that contained Shigella and four of 314 stools from which Shigella and enteroinvasive E. coli were not isolated. The use of DNA probes allows specimens to be collected in remote areas with a minimum amount of equipment and technical expertise so that they can be easily transported to a central laboratory for further processing.

Cytosolic Access of Intracellular Bacterial Pathogens: The Shigella Paradigm.

Science.gov (United States)

Mellouk, Nora; Enninga, Jost

2016-01-01

Shigella is a Gram-negative bacterial pathogen, which causes bacillary dysentery in humans. A crucial step of Shigella infection is its invasion of epithelial cells. Using a type III secretion system, Shigella injects several bacterial effectors ultimately leading to bacterial internalization within a vacuole. Then, Shigella escapes rapidly from the vacuole, it replicates within the cytosol and spreads from cell-to-cell. The molecular mechanism of vacuolar rupture used by Shigella has been studied in some detail during the recent years and new paradigms are emerging about the underlying molecular events. For decades, bacterial effector proteins were portrayed as main actors inducing vacuolar rupture. This includes the effector/translocators IpaB and IpaC. More recently, this has been challenged and an implication of the host cell in the process of vacuolar rupture has been put forward. This includes the bacterial subversion of host trafficking regulators, such as the Rab GTPase Rab11. The involvement of the host in determining bacterial vacuolar integrity has also been found for other bacterial pathogens, particularly for Salmonella. Here, we will discuss our current view of host factor and pathogen effector implications during Shigella vacuolar rupture and the steps leading to it.

Investigation of lethal and mutagenetic effects of UV-light on Salmonella currying wild and mutant alleles of lex A gene of Escherichia coli in the Salmonella genome

International Nuclear Information System (INIS)

Andreeva, I.V.; Tiganova, I.G.; Skavronskaya, A.G.

1981-01-01

Inheritance of LexA-gene of Escherichia coli- by Salmonella takes place during intergeneric trunsduction cross of Escherichia coli and Salmonella typhimurium. The presence of LexA-E. coli gene-did not eliminate earlier revealed peculiarity consisting in the absence of UV-induced mutagenesis in most of studied salmonollosis strains. So it is shown that the absence of UV mutagenesis in Salmonella does not result from mutation in LexA-gene. Inheritance of pKM101 by LexA-hybrid provides pronounced UV mutability and protective effect. Inheritance of this plasmid by LexA-hybrid did not result in the appearance of capability for UV-induced mutagenesis and improving UV resistance of bacteria. Thus the plasmids effect on repair and mutagenesis in Salmonella, the same as in E. coli, reveals in LexA-phenotype [ru

The development of methods for the detection of Salmonella in chickens by a combination of immunomagnetic separation and PCRs.

Science.gov (United States)

Dai, Fengying; Zhang, Miao; Xu, Dixin; Yang, Yin; Wang, Jiaxiao; Li, Mingzhen; Du, Meihong

2017-11-01

Micro- and nanoimmunomagnetic beads (MIMBs and NIMBs) used for immunomagnetic separation (IMS) with PCR were studied for the rapid detection of Salmonella. The capture efficiency of the two different IMBs was evaluated by a conventional plate counting method, and the binding pattern was studied using scanning electron microscopy. The specificity of the IMBs was tested with Salmonella, Shigella flexneri, enterohemorrhagic Escherichia coli O157:H7, and Listeria monocytogenes. By comparing the pre-enrichment IMS and the IMS enrichment steps with a 5.5-H enrichment time, this study developed a rapid and sensitive method for the detection of Salmonella in chicken. The method was implemented by IMS enrichment and PCR with MIMBs and NIMBs, with a total analysis time of 8 H. We showed that the method was sensitive based on NIMBs with a detection limit of 10° CFU for Salmonella in 25 g of chicken. © 2016 International Union of Biochemistry and Molecular Biology, Inc.

Detecção de fatores de virulência de Escherichia coli e análise de Salmonella spp. em psitacídeos Detection of virulence factors in Escherichia coli and analysis of Salmonella spp. in psittacines

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Isadora M. de O. Corrêa

2013-02-01

Full Text Available A flora entérica dos psitacídeos é composta principalmente por bactérias Gram positivas. Bactérias Gram negativas, como Escherichia coli e Salmonella spp., apresentam elevado potencial patogênico, sendo consideradas indicativo de problemas de manejo, que poderão culminar em manifestação de doenças em decorrência de fatores estressantes, dietas deficientes e superlotação, combinados com alta carga bacteriana no ambiente. O objetivo deste trabalho foi avaliar a presença de Salmonella spp., Escherichia coli e os fatores de virulência dos genes iss e iutA dos isolados de E. coli. Analisou-se um total de 44 amostras provenientes de psitacídeos criados em cativeiro, sendo estas 15 fragmentos de órgãos de aves submetidas a exame de necropsia e também 29 amostras de swabs de cloaca e inglúvio de papagaios-charão (Amazona pretrei criados em cativeiro. Nenhuma amostra foi positiva para Salmonella spp. Nas amostras de E. coli detectou-se ambos os fatores de virulência pesquisados.The enteric flora of psittacines is mainly composed of Gram positive bacteria. Gram negative bacteria, like Escherichia coli and Salmonella spp., have a high pathogenic potential and can be considerate as an indicative of management problems that may culminate in disease manifestation due to stress factors, poor diets and overcrowding, in combination with a high bacterial load on the environment. The objective of this study was evaluated the presence of Salmonella spp., Escherichia coli and the virulence genes iss and iutA from E. coli isolates. Forty-four samples were analyzed from psittacines living in captivity, which fifteen samples were from organs fragments of necropsied birds, and twenty-nine were from cloacal and crop swabs of red-spectacled parrots (Amazona pretrei keeping in captivity. No samples were positive for Salmonella spp. In the samples in which E. coli was detected, both virulence factors (genes iss and iutA were present.

[Etiological surveillance and analysis of infectious diarrhea in Beijing in year 2010].

Science.gov (United States)

Huang, Fang; Deng, Ying; Qu, Mei; Liu, Gui-Rong; Liu, Yuan; Zhang, Xin; Li, Jie; Yan, Han-Qiu; Gao, Zhi-Yong; Liu, Bai-Wei; Li, Xi-Tai; Li, Xin-Yu

2011-09-01

To explore the pathogenic form, epidemic features and serotype distribution of the pathogenic bacteria causing infectious diarrhea in Beijing. A total of 2118 samples of rectal swabs and stool specimens of diarrheal patients were collected from 6 surveillant intestinal tract clinics during the period between April and October, 2010. Enteric multiple pathogens including Vibrio cholerae, Vibrio parahaemolyticus, Salmonella, Shigella and diarrheagenic Escherichia coli were detected by the isolation culture, biochemical identification and serotyping methods. The population distribution, temporal distribution and serotype distribution of the above pathogenic bacteria were analyzed by descriptive statistical methods. 478 strains isolated from the total 2118 specimens were positive for pathogen detection, accounting to 22.6%. Among the 478 strains of pathogenic bacteria, Shigella accounting for 40.8% (195/478) was the most frequent pathogen, followed by Vibrio parahaemolyticus accouting for 23.8% (114/478), Salmonella accounting for 19.0% (91/478) and diarrheagenic Escherichia coli accounting for 4.8% (23/478). Enteric pathogenic bacteria spread mainly among adults aging between 20 and 39; and the distribution was different among different age groups, while the highest detected rate was in 30 - 39 age group, accounting for 27.2% (92/338). The detected rate of pathogenic bacteria showed evident seasonal variations, with a peak from July to October, whose detected rates were 23.5% (114/486), 32.8% (176/536), 36.1% (90/249) and 25.9% (29/112) respectively. The detected rates in other months were all under 16.0%. Shigella Sonnei was the dominant serotype, accounting for 83.1% (162/195). O3:K6 was the dominant serotype among Vibrio parahaemolyticus, accounting for 63.2% (72/114). Salmonella Enteritidis and Salmonella Typhimurium were dominant serotypes among Salmonella, accounting for 13.2% (12/91) and 12.1% (11/91) separately. Enterpathogenic Escherichia coli and

Cytosolic access of intracellular bacterial pathogens: the Shigella paradigm

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Nora eMellouk

2016-04-01

Full Text Available Shigella is a Gram-negative bacterial pathogen, which causes bacillary dysentery in humans. A crucial step of Shigella infection is its invasion of epithelial cells. Using a type III secretion system, Shigella injects several bacterial effectors ultimately leading to bacterial internalization within a vacuole. Then, Shigella escapes rapidly from the vacuole, it replicates within the cytosol and spreads from cell-to-cell. The molecular mechanism of vacuolar rupture used by Shigella has been studied in some detail during the recent years and new paradigms are emerging about the underlying molecular events. For decades, bacterial effector proteins were portrayed as main actors inducing vacuolar rupture. This includes the effector/translocators IpaB and IpaC. More recently, this has been challenged and an implication of the host cell in the process of vacuolar rupture has been put forward. This includes the bacterial subversion of host trafficking regulators, such as the Rab GTPase Rab11. The involvement of the host in determining bacterial vacuolar integrity has also been found for other bacterial pathogens, particularly for Salmonella. Here, we will discuss our current view of host factor and pathogen effector implications during Shigella vacuolar rupture and the steps leading to it.

Identificação de cepas de Escherichia coli enteropatogênicas em amostras de fezes, por reação de imunofluorescência direta Identification of strains of Escherichia coli in stool samples by direct fluorescent antibody tests

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Ieda Maria Longo

1980-06-01

Full Text Available Foi realizado diagnóstico etiológico de casos de diarréia aguda em 121 pacientes internados na Clínica Pediátrica do Hospital da Santa Casa de São Paulo, Brasil. Foram utilizados os métodos bacteriológico clássico e de reação de imunofluorescência direta para a identificação de cepas de Escherichia coli enteropatogênicas: para estudo da sensibilidade das cepas de Escherichia coli isoladas, a diferentes antibióticos, foi usado o método de Concentração Inibitória Minima (CIM. Dos 56 casos positivos, 89,3% correspondiam a diferentes sorotipos enteropatogênicos de Escherichia coli, quando utilizada a técnica de imunofluorescência direta. O método bacteriológico clássico revelou ainda, nos 121 casos examinados, 4 cepas de Salmonella e 2 de Shigella. No estudo da CIM verificou-se maior sensibilidade das cepas de Escherichia coli enteropatogênicas estudadas à Gentamicina e Amikacina, do que aos outros antibióticos.A study of 121 patients with acute diarrhea was made at the Pediatric Clinic of the Santa Casa de S. Paulo (the S. Paulo Charity Hospital. Etiological diagnosis of 121 cases was carried out through the classical bacteriological method and direct fluorescent antibody tests for the identification of E. coli. The antibiotic sensitivity of these bacteria to different antimicrobials was determined by the Minimum Inhibitory Concentration (MIC method. Fifty-six positive cases were found; 89.3% of which corresponded to different serotypes of enter o pathogenic E. coli (89.3%, when the direct fluorescent antibody test was used. The classic bacteriological method bared four Salmonella strains and two Shigella. The MIC showed the E. coli to be more sensitive to Gentamicin and Amikacin than to other antibiotics.

Defining the phylogenomics of Shigella species: a pathway to diagnostics.

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Sahl, Jason W; Morris, Carolyn R; Emberger, Jennifer; Fraser, Claire M; Ochieng, John Benjamin; Juma, Jane; Fields, Barry; Breiman, Robert F; Gilmour, Matthew; Nataro, James P; Rasko, David A

2015-03-01

Shigellae cause significant diarrheal disease and mortality in humans, as there are approximately 163 million episodes of shigellosis and 1.1 million deaths annually. While significant strides have been made in the understanding of the pathogenesis, few studies on the genomic content of the Shigella species have been completed. The goal of this study was to characterize the genomic diversity of Shigella species through sequencing of 55 isolates representing members of each of the four Shigella species: S. flexneri, S. sonnei, S. boydii, and S. dysenteriae. Phylogeny inferred from 336 available Shigella and Escherichia coli genomes defined exclusive clades of Shigella; conserved genomic markers that can identify each clade were then identified. PCR assays were developed for each clade-specific marker, which was combined with an amplicon for the conserved Shigella invasion antigen, IpaH3, into a multiplex PCR assay. This assay demonstrated high specificity, correctly identifying 218 of 221 presumptive Shigella isolates, and sensitivity, by not identifying any of 151 diverse E. coli isolates incorrectly as Shigella. This new phylogenomics-based PCR assay represents a valuable tool for rapid typing of uncharacterized Shigella isolates and provides a framework that can be utilized for the identification of novel genomic markers from genomic data. Copyright © 2015, American Society for Microbiology. All Rights Reserved.

In vitro adherence patterns of Shigella serogroups to bovine recto-anal junction squamous epithelial (RSE) cells are similar to those of Escherichia coli O157

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The aim of this study was to determine whether Shigella species, which are human gastrointestinal pathogens, can adhere to cattle recto-anal junction squamous epithelial (RSE) cells using a recently standardized adherence assay, and to compare their adherence patterns to that of Escherichia coli O15...

Plasmid-Mediated Quinolone Resistance in Shigella flexneri Isolated From Macaques

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Anthony J. Mannion

2018-03-01

Full Text Available Non-human primates (NHPs for biomedical research are commonly infected with Shigella spp. that can cause acute dysentery or chronic episodic diarrhea. These animals are often prophylactically and clinically treated with quinolone antibiotics to eradicate these possible infections. However, chromosomally- and plasmid-mediated antibiotic resistance has become an emerging concern for species in the family Enterobacteriaceae. In this study, five individual isolates of multi-drug resistant Shigella flexneri were isolated from the feces of three macaques. Antibiotic susceptibility testing confirmed resistance or decreased susceptibility to ampicillin, amoxicillin-clavulanic acid, cephalosporins, gentamicin, tetracycline, ciprofloxacin, enrofloxacin, levofloxacin, and nalidixic acid. S. flexneri isolates were susceptible to trimethoprim-sulfamethoxazole, and this drug was used to eradicate infection in two of the macaques. Plasmid DNA from all isolates was positive for the plasmid-encoded quinolone resistance gene qnrS, but not qnrA and qnrB. Conjugation and transformation of plasmid DNA from several S. flexneri isolates into antibiotic-susceptible Escherichia coli strains conferred the recipients with resistance or decreased susceptibility to quinolones and beta-lactams. Genome sequencing of two representative S. flexneri isolates identified the qnrS gene on a plasmid-like contig. These contigs showed >99% homology to plasmid sequences previously characterized from quinolone-resistant Shigella flexneri 2a and Salmonella enterica strains. Other antibiotic resistance genes and virulence factor genes were also identified in chromosome and plasmid sequences in these genomes. The findings from this study indicate macaques harbor pathogenic S. flexneri strains with chromosomally- and plasmid-encoded antibiotic resistance genes. To our knowledge, this is the first report of plasmid-mediated quinolone resistance in S. flexneri isolated from NHPs and warrants

Identification of novel substrates of Shigella T3SA through analysis of its virulence plasmid-encoded secretome

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Pinaud, Laurie; Ferrari, Mariana L.; Friedman, Robin; Jehmlich, Nico; von Bergen, Martin; Phalipon, Armelle; Sansonetti, Philippe J.

2017-01-01

Many human Gram-negative bacterial pathogens express a Type Three Secretion Apparatus (T3SA), including among the most notorious Shigella spp., Salmonella enterica, Yersinia enterocolitica and enteropathogenic Escherichia coli (EPEC). These bacteria express on their surface multiple copies of the T3SA that mediate the delivery into host cells of specific protein substrates critical to pathogenesis. Shigella spp. are Gram-negative bacterial pathogens responsible for human bacillary dysentery. The effector function of several Shigella T3SA substrates has largely been studied but their potential cellular targets are far from having been comprehensively delineated. In addition, it is likely that some T3SA substrates have escaped scrutiny as yet. Indeed, sequencing of the virulence plasmid of Shigella flexneri has revealed numerous open reading frames with unknown functions that could encode additional T3SA substrates. Taking advantage of label-free mass spectrometry detection of proteins secreted by a constitutively secreting strain of S. flexneri, we identified five novel substrates of the T3SA. We further confirmed their secretion through the T3SA and translocation into host cells using β-lactamase assays. The coding sequences of two of these novel T3SA substrates (Orf13 and Orf131a) have a guanine-cytosine content comparable to those of T3SA components and effectors. The three other T3SA substrates identified (Orf48, Orf86 and Orf176) have significant homology with antitoxin moieties of type II Toxin-Antitoxin systems usually implicated in the maintenance of low copy plasmids. While Orf13 and Orf131a might constitute new virulence effectors contributing to S. flexneri pathogenicity, potential roles for the translocation into host cells of antitoxins or antitoxin-like proteins during Shigella infection are discussed. PMID:29073283

Identification of novel substrates of Shigella T3SA through analysis of its virulence plasmid-encoded secretome.

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Laurie Pinaud

Full Text Available Many human Gram-negative bacterial pathogens express a Type Three Secretion Apparatus (T3SA, including among the most notorious Shigella spp., Salmonella enterica, Yersinia enterocolitica and enteropathogenic Escherichia coli (EPEC. These bacteria express on their surface multiple copies of the T3SA that mediate the delivery into host cells of specific protein substrates critical to pathogenesis. Shigella spp. are Gram-negative bacterial pathogens responsible for human bacillary dysentery. The effector function of several Shigella T3SA substrates has largely been studied but their potential cellular targets are far from having been comprehensively delineated. In addition, it is likely that some T3SA substrates have escaped scrutiny as yet. Indeed, sequencing of the virulence plasmid of Shigella flexneri has revealed numerous open reading frames with unknown functions that could encode additional T3SA substrates. Taking advantage of label-free mass spectrometry detection of proteins secreted by a constitutively secreting strain of S. flexneri, we identified five novel substrates of the T3SA. We further confirmed their secretion through the T3SA and translocation into host cells using β-lactamase assays. The coding sequences of two of these novel T3SA substrates (Orf13 and Orf131a have a guanine-cytosine content comparable to those of T3SA components and effectors. The three other T3SA substrates identified (Orf48, Orf86 and Orf176 have significant homology with antitoxin moieties of type II Toxin-Antitoxin systems usually implicated in the maintenance of low copy plasmids. While Orf13 and Orf131a might constitute new virulence effectors contributing to S. flexneri pathogenicity, potential roles for the translocation into host cells of antitoxins or antitoxin-like proteins during Shigella infection are discussed.

Surface display of Salmonella epitopes in Escherichia coli and Staphylococcus carnosus.

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Nhan, Nguyen Thanh; Gonzalez de Valdivia, Ernesto; Gustavsson, Martin; Hai, Truong Nam; Larsson, Gen

2011-04-11

Salmonella enterica serotype Enteritidis (SE) is considered to be one of the most potent pathogenic Salmonella serotypes causing food-borne disease in humans. Since a live bacterial vaccine based on surface display of antigens has many advantages over traditional vaccines, we have studied the surface display of the SE antigenic proteins, H:gm and SefA in Escherichia coli by the β-autotransporter system, AIDA. This procedure was compared to protein translocation in Staphylococcus carnosus, using a staphylococci hybrid vector earlier developed for surface display of other vaccine epitopes. Both SefA and H:gm were translocated to the outer membrane in Escherichia coli. SefA was expressed to full length but H:gm was shorter than expected, probably due to a proteolytic cleavage of the N-terminal during passage either through the periplasm or over the membrane. FACS analysis confirmed that SefA was facing the extracellular environment, but this could not be conclusively established for H:gm since the N-terminal detection tag (His6) was cleaved off. Polyclonal salmonella antibodies confirmed the sustained antibody-antigen binding towards both proteins. The surface expression data from Staphylococcus carnosus suggested that the H:gm and SefA proteins were transported to the cell wall since the detection marker was displayed by FACS analysis. Apart from the accumulated knowledge and the existence of a wealth of equipment and techniques, the results indicate the selection of E. coli for further studies for surface expression of salmonella antigens. Surface expression of the full length protein facing the cell environment was positively proven by standard analysis, and the FACS signal comparison to expression in Staphylococcus carnosus shows that the distribution of the surface protein on each cell was comparatively very narrow in E. coli, the E. coli outer membrane molecules can serve as an adjuvant for the surface antigenic proteins and multimeric forms of the SefA protein

Rapid and reliable discrimination between Shigella species and Escherichia coli using MALDI-TOF mass spectrometry.

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Paauw, Armand; Jonker, Debby; Roeselers, Guus; Heng, Jonathan M E; Mars-Groenendijk, Roos H; Trip, Hein; Molhoek, E Margo; Jansen, Hugo-Jan; van der Plas, Jan; de Jong, Ad L; Majchrzykiewicz-Koehorst, Joanna A; Speksnijder, Arjen G C L

2015-01-01

E. coli-Shigella species are a cryptic group of bacteria in which the Shigella species are distributed within the phylogenetic tree of E. coli. The nomenclature is historically based and the discrimination of these genera developed as a result of the epidemiological need to identify the cause of shigellosis, a severe disease caused by Shigella species. For these reasons, this incorrect classification of shigellae persists to date, and the ability to rapidly characterize E. coli and Shigella species remains highly desirable. Until recently, existing matrix-assisted laser desorption/ionization-time of flight mass spectrometry (MALDI-TOF MS) assays used to identify bacteria could not discriminate between E. coli and Shigella species. Here we present a rapid classification method for the E. coli-Shigella phylogroup based on MALDI-TOF MS which is supported by genetic analysis. E. coli and Shigella isolates were collected and genetically characterized by MLVA. A custom reference library for MALDI-TOF MS that represents the genetic diversity of E. coli and Shigella strains was developed. Characterization of E. coli and Shigella species is based on an approach with Biotyper software. Using this reference library it was possible to distinguish between Shigella species and E. coli. Of the 180 isolates tested, 94.4% were correctly classified as E. coli or shigellae. The results of four (2.2%) isolates could not be interpreted and six (3.3%) isolates were classified incorrectly. The custom library extends the existing MALDI-TOF MS method for species determination by enabling rapid and accurate discrimination between Shigella species and E. coli. Copyright © 2015 Elsevier GmbH. All rights reserved.

Escherichia coli and Salmonella ser. Saintpaul natural co-infection in a free-living ruddy ground dove (Columbina talpacoti: a case report

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W.G.A. Bezerra

Full Text Available ABSTRACT This study reports a co-infection of Escherichia coli and Salmonella in a free-living ruddy ground dove (Columbina talpacoti received at the Laboratory of Ornithological Studies of the State University of Ceará, Brazil. The bird presented diarrhea, leg paralysis and anorexia, and died shortly after. Necropsy was then performed and samples from lung, kidney, liver and intestine were collected for microbiological and histopathological analyses. Escherichia coli was isolated from cloacal swab, lung and kidney samples. Salmonella ser. Saintpaul was identified in liver and spleen samples. Escherichia coli isolates were tested for the presence of eight diagnostic genes for diarrheagenic pathotypes (STEC, ETEC, EPEC, EIEC, EAEC with conventional polymerase chain reaction (PCR. EAEC was detected in the lung and kidney, and STEC in the intestine. In conclusion, Columbina talpacoti is susceptible to enteroaggregative Escherichia coli and Salmonella ser. Saintpaul infection, which may have public health implications.

Shigella Diversity and Changing Landscape: Insights for the Twenty-First Century

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Anderson, Mark; Sansonetti, Philippe J.; Marteyn, Benoit S.

2016-01-01

Shigella is a pathovar of Escherichia coli comprising four groups, Shigella flexneri, Shigella sonnei, Shigella dysenteriae, and Shigella boydii, each of them, with the exception of S.sonnei, comprising several serotypes. Shigella accounts for the majority of dysentery causing infections occurring world-wide each year. Recent advancements in the Shigella field have led to a better understanding of the molecular mechanisms underlying host epithelial cell invasion and immune cell function manipulation, mainly using S. flexneri as a model. Host-cell invasion is the final step of the infection process, as Shigella's virulence strategy relies also on its ability to survive hostile conditions during its journey through the gastro-intestinal tract, to compete with the host microbiota and to cross the intestinal mucus layer. Hence, the diversity of the virulence strategies among the different Shigella species has not yet been deeply investigated, which might be an important step to understand the epidemiological spreading of Shigella species worldwide and a key aspect for the validation of novel vaccine candidates. The recent development of high-throughput screening and sequencing methods will facilitate these complex comparison studies. In this review we discuss several of the major avenues that the Shigella research field has taken over the past few years and hopefully gain some insights into the questions that remain surrounding this important human pathogen. PMID:27148494

Development of a Flow Cytometry-Based Method for Rapid Detection of Escherichia coli and Shigella Spp. Using an Oligonucleotide Probe

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Xue, Yong; Wilkes, Jon G.; Moskal, Ted J.; Williams, Anna J.; Cooper, Willie M.; Nayak, Rajesh; Rafii, Fatemeh; Buzatu, Dan A.

2016-01-01

Standard methods to detect Escherichia coli contamination in food use the polymerase chain reaction (PCR) and agar culture plates. These methods require multiple incubation steps and take a long time to results. An improved rapid flow-cytometry based detection method was developed, using a fluorescence-labeled oligonucleotide probe specifically binding a16S rRNA sequence. The method positively detected 51 E. coli isolates as well as 4 Shigella species. All 27 non-E. coli strains tested gave negative results. Comparison of the new genetic assay with a total plate count (TPC) assay and agar plate counting indicated similar sensitivity, agreement between cytometry cell and colony counts. This method can detect a small number of E.coli cells in the presence of large numbers of other bacteria. This method can be used for rapid, economical, and stable detection of E. coli and Shigella contamination in the food industry and other contexts. PMID:26913737

Development of a Flow Cytometry-Based Method for Rapid Detection of Escherichia coli and Shigella Spp. Using an Oligonucleotide Probe.

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Yong Xue

Full Text Available Standard methods to detect Escherichia coli contamination in food use the polymerase chain reaction (PCR and agar culture plates. These methods require multiple incubation steps and take a long time to results. An improved rapid flow-cytometry based detection method was developed, using a fluorescence-labeled oligonucleotide probe specifically binding a16S rRNA sequence. The method positively detected 51 E. coli isolates as well as 4 Shigella species. All 27 non-E. coli strains tested gave negative results. Comparison of the new genetic assay with a total plate count (TPC assay and agar plate counting indicated similar sensitivity, agreement between cytometry cell and colony counts. This method can detect a small number of E.coli cells in the presence of large numbers of other bacteria. This method can be used for rapid, economical, and stable detection of E. coli and Shigella contamination in the food industry and other contexts.

Characterization of antimicrobial activity in Kombucha fermentation

NARCIS (Netherlands)

Sreeramulu, G.; Zhu, Y.; Knol, W.

2001-01-01

Fermented tea drink, Kombucha, can inhibit the growth of Shigella sonnei, Escherichia coli, Salmonella enteritidis and Salmonella typhimurium. Several metabolites were analyzed every two days during a 14-day Kombucha fermentation. Levels of acetic acid and gluconic acid were found to increase with

Elimination of Escherichia coli and Salmonella in Clam by Using Zeolite in a Station of Depuration.

Science.gov (United States)

Gdoura, Morsi; Sellami, Hanen; Khannous, Lamia; Ketata, Najib; Neila, Idriss Ben; Traore, Al Ibrahim; Chekir, Zouhair; Gdoura, Radhouane

2017-09-01

  The application of natural zeolite for water and wastewater treatment has been carried out and is still a promising technique in environmental cleaning processes. Natural zeolite can be used to improve the purification process of clams (Ruditapes decussatus). Thus, our study aimed at improving the clam purification system in order to reduce Escherichia coli and eliminate Salmonella in samples artificially contaminated with this bacterium using a natural zeolite to replace the biological filter. The results showed that zeolite used in a depuration system improved the clam purification process. Moreover, natural zeolite exhibited high performance in the adsorption of bacteria and allowed to reduce the Escherichia coli abundance in 24 h, thus ensuring purified clams conformity with the ISO 16649-3 standard. These results indicate the beneficial effects of using zeolite in the adsorption of bacteria and the reduction in the abundance of Escherichia coli and set the Salmonella from marine organisms.

Effect of the oyster contaminated in Salmonella typhimurium and Escherichia coli; Efecto del ostion contaminado en Salmonella typhimurium y Escherichia coli

Energy Technology Data Exchange (ETDEWEB)

Brena V, M

1992-08-15

In this work the effect of the oyster contaminated in the species of bacteria better studied by its genetic composition these are the Salmonella typhimurium and the Escherichia coli and that its have been starting point for later radiobiological studies in superior organisms. Of this its have arisen two general lines. The research about the genotoxic effect of substances or mixtures in bacteria with the collaboration of the groups of Drosophila and X-ray Fluorescence analysis and on the other hand the study of the low doses of radiation also in bacteria. It is also treated the topic about the genetic effect of aromatic hydrocarbons in different biological systems. (Author)

Effect of Antimicrobial Dosage Regimen on Salmonella and Escherichia coli Isolates from Feeder Swine▿

Science.gov (United States)

Wagner, Bruce A.; Straw, Barbara E.; Fedorka-Cray, Paula J.; Dargatz, David A.

2008-01-01

A body of evidence exists that suggests that antimicrobial use in food animals leads to resistance in both pathogenic and commensal bacteria. This study focused on the impact of three different antimicrobial regimes (low-level continuous, pulse, and no antimicrobial) for two antimicrobials (chlortetracycline and tylosin) on the presence of Salmonella spp. and on the prevalence of antimicrobial resistance of both Salmonella spp. and nonspecific Escherichia coli in fecal samples from feeder swine. The prevalence of fecal samples positive for Salmonella spp. significantly decreased between the samples taken at feeder placement compared to samples taken when the animals were close to market weight. Differences in resistance of Salmonella spp. did not appear to be influenced by dosing treatment including the control. Analysis of antimicrobial resistance examining both susceptibility and resistance, as well as MIC outcomes, demonstrated that only resistance to cephalothin increased in E. coli under the pulse chlortetracycline treatment. These results suggest that the dosing regimes examined in this study did not lead to an increase in either the prevalence of Salmonella spp. or the prevalence of antimicrobial resistance in isolates of Salmonella spp. or E. coli. PMID:18223115

Enterobacteriaceae in dehydrated powdered infant formula manufactured in Indonesia and Malaysia.

Science.gov (United States)

Estuningsih, Sri; Kress, Claudia; Hassan, Abdulwahed A; Akineden, Omer; Schneider, Elisabeth; Usleber, Ewald

2006-12-01

To determine the occurrence of Salmonella and Shigella in infant formula from Southeast Asia, 74 packages of dehydrated powdered infant follow-on formula (recommended age, > 4 months) from five different manufacturers, four from Indonesia and one from Malaysia, were analyzed. None of the 25-g test portions yielded Salmonella or Shigella. However, further identification of colonies growing on selective media used for Salmonella and Shigella detection revealed the frequent occurrence of several other Enterobacteriaceae species. A total of 35 samples (47%) were positive for Enterobacteriaceae. Ten samples (13.5%) from two Indonesian manufacturers yielded Enterobacter sakazakii. Other Enterobacteriaceae isolated included Pantoea spp. (n = 12), Escherichia hermanii (n = 10), Enterobacter cloacae (n = 8), Klebsiella pneumoniae subsp. pneumoniae (n = 3), Citrobacter spp. (n = 2), Serratia spp. (n = 2), and Escherichia coli (n = 2). To our knowledge, this is the first report to describe the contamination of dehydrated powdered infant formula from Indonesia with E. sakazakii and several other Enterobacteriaceae that could be opportunistic pathogens. Improper preparation and conservation of these products could result in a health risk for infants in Indonesia.

The use of comparative genomic hybridization to characterize genome dynamics and diversity among the serotypes of Shigella

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Sun Meisheng

2006-08-01

Full Text Available Abstract Background Compelling evidence indicates that Shigella species, the etiologic agents of bacillary dysentery, as well as enteroinvasive Escherichia coli, are derived from multiple origins of Escherichia coli and form a single pathovar. To further understand the genome diversity and virulence evolution of Shigella, comparative genomic hybridization microarray analysis was employed to compare the gene content of E. coli K-12 with those of 43 Shigella strains from all lineages. Results For the 43 strains subjected to CGH microarray analyses, the common backbone of the Shigella genome was estimated to contain more than 1,900 open reading frames (ORFs, with a mean number of 726 undetectable ORFs. The mosaic distribution of absent regions indicated that insertions and/or deletions have led to the highly diversified genomes of pathogenic strains. Conclusion These results support the hypothesis that by gain and loss of functions, Shigella species became successful human pathogens through convergent evolution from diverse genomic backgrounds. Moreover, we also found many specific differences between different lineages, providing a window into understanding bacterial speciation and taxonomic relationships.

Genetic Transfer of Salmonella typhimurium and Escherichia coli Lipopolysaccharide Antigens to Escherichia coli K-12

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Jones, Randall T.; Koeltzow, Donald E.; Stocker, B. A. D.

1972-01-01

Escherichia coli K-12 ϰ971 was crossed with a smooth Salmonella typhimurium donor, HfrK6, which transfers early the ilv-linked rfa region determining lipopolysaccharide (LPS) core structure. Two ilv+ hybrids differing in their response to the LPS-specific phages FO and C21 were then crossed with S. typhimurium HfrK9, which transfers early the rfb gene cluster determining O repeat unit structure. Most recombinants selected for his+ (near rfb) were agglutinated by Salmonella factor 4 antiserum. Transfer of an F′ factor (FS400) carrying the rfb–his region of S. typhimurium to the same two ilv+ hybrids gave similar results. LPS extracted from two ilv+,his+, factor 4-positive hybrids contained abequose, the immunodominant sugar for factor 4 specificity. By contrast, his+ hybrids obtained from ϰ971 itself by similar HfrK9 and F′FS400 crosses were not agglutinated by factor 4 antiserum, indicating that the parental E. coli ϰ971 does not have the capacity to attach Salmonella O repeat units to its LPS core. It is concluded that the Salmonella rfb genes are expressed only in E. coli ϰ971 hybrids which have also acquired ilv-linked genes (presumably rfa genes affecting core structure or O-translocase ability, or both) from a S. typhimurium donor. When E. coli ϰ971 was crossed with a smooth E. coli donor, Hfr59, of serotype O8, which transfers his early, most his+ recombinants were agglutinated by E. coli O8 antiserum and lysed by the O8-specific phage, Ω8. This suggests that, although the parental E. coli K-12 strain ϰ971 cannot attach Salmonella-specific repeat units to its LPS core, it does have the capacity to attach E. coli O8-specific repeat units. PMID:4559827

Shigella Infections

Science.gov (United States)

... Staying Safe Videos for Educators Search English Español Shigella Infections KidsHealth / For Parents / Shigella Infections What's in ... Doctor Print en español Infecciones por Shigella About Shigella Shigella are bacteria that can infect the digestive ...

Shigella infection of intestinal epithelium and circumvention of the host innate defense system.

Science.gov (United States)

Ashida, Hiroshi; Ogawa, Michinaga; Mimuro, Hitomi; Sasakawa, Chihiro

2009-01-01

Shigella, Gram-negative bacteria closely related to Escherichia coli, are highly adapted human pathogens that cause bacillary dysentery. Although Shigella have neither adherence factors nor flagella required for attaching or accessing the intestinal epithelium, Shigella are capable of colonizing the intestinal epithelium by exploiting epithelial-cell functions and circumventing the host innate immune response. During Shigella infection, they deliver many numbers of effectors through the type III secretion system into the surrounding space and directly into the host-cell cytoplasm. The effectors play pivotal roles from the onset of bacterial infection through to the establishment of the colonization of the intestinal epithelium, such as bacterial invasion, intracellular survival, subversion of the host immune defense response, and maintenance of the infectious foothold. These examples suggest that Shigella have evolved highly sophisticated infectious and intracellular strategies to establish replicative niches in the intestinal epithelium.

Gamma radiation in the reduction of Salmonella spp. inoculated on minimally processed watercress (Nasturtium officinalis)

International Nuclear Information System (INIS)

Martins, C.G.; Behrens, J.H.; Destro, M.T.; Franco, B.D.G.M.; Vizeu, D.M.; Hutzler, B.; Landgraf, M.

2004-01-01

Consumer attitudes towards foods have changed in the last two decades increasing requirements for freshlike products. Consequently, less extreme treatments or additives are being required. Minimally processed foods have freshlike characteristics and satisfy this new consumer demand. Besides freshness, the minimally processing also provide convenience required by the market. Salad vegetables can be source of pathogen such as Salmonella, Escherichia coli O157:H7, Shigella spp. The minimal processing does not reduce the levels of pathogenic microorganisms to safe levels. Therefore, this study was carried out in order to improve the microbiological safety and the shelf-life of minimally processed vegetables using gamma radiation. Minimally processed watercress inoculated with a cocktail of Salmonella spp was exposed to 0.0, 0.2, 0.5, 0.7, 1.0, 1.2 and 1.5 kGy. Irradiated samples were diluted 1:10 in saline peptone water and plated onto tryptic soy agar that were incubated at 37 deg. C/24 h. D 10 values for Salmonella spp. inoculated in watercress varied from 0.29 to 0.43 kGy. Therefore, a dose of 1.7 kGy will reduce Salmonella population in watercress by 4 log 10 . The shelf-life was increased by 1 1/2 day when the product was exposed to 1 kGy

A rapid Salmonella detection method involving thermophilic helicase-dependent amplification and a lateral flow assay.

Science.gov (United States)

Du, Xin-Jun; Zhou, Tian-Jiao; Li, Ping; Wang, Shuo

2017-08-01

Salmonella is a major foodborne pathogen that is widespread in the environment and can cause serious human and animal disease. Since conventional culture methods to detect Salmonella are time-consuming and laborious, rapid and accurate techniques to detect this pathogen are critically important for food safety and diagnosing foodborne illness. In this study, we developed a rapid, simple and portable Salmonella detection strategy that combines thermophilic helicase-dependent amplification (tHDA) with a lateral flow assay to provide a detection result based on visual signals within 90 min. Performance analyses indicated that the method had detection limits for DNA and pure cultured bacteria of 73.4-80.7 fg and 35-40 CFU, respectively. Specificity analyses showed no cross reactions with Escherichia coli, Staphylococcus aureus, Listeria monocytogenes, Enterobacter aerogenes, Shigella and Campylobacter jejuni. The results for detection in real food samples showed that 1.3-1.9 CFU/g or 1.3-1.9 CFU/mL of Salmonella in contaminated chicken products and infant nutritional cereal could be detected after 2 h of enrichment. The same amount of Salmonella in contaminated milk could be detected after 4 h of enrichment. This tHDA-strip can be used for the rapid detection of Salmonella in food samples and is particularly suitable for use in areas with limited equipment. Copyright © 2017 Elsevier Ltd. All rights reserved.

Salmonelose humana e animal em Araraquara, S. Paulo: prevalência de Shigella em casos humanos

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Deise Pasetto Falcão

1975-10-01

Full Text Available Admitida uma baixa freqüência de Salmonella com base em verificações anteriores em casos de diarréia aguda na cidade de Araraquara, S. Paulo, realizou-se uma investigação com um esquema ampliado para o isolamento de Salmonella a partir de 47 casos de gastroenterite infantil, 51 amostras de fezes de animais e 50 amostras de carnes e vísceras de animais destinados a alimentação. Foram isoladas Salmonella em 6,3% dos casos de gastroenterite, 5,8% de fezes de animais e 8,0% de amostras de alimentos, nesse último caso verificando-se a contaminação exclusivamente em fígado de porco, senão negativos todos os resultados de carne e vísceras de bovinos. Os sorotipos isolados corresponderam, em ordem decrescente de freqüência a S. anatum, S. derby e S. daytona. Nas gastroenterites infantis a freqüência de Shigella (12,7% foi duas vezes superior à de Salmonella (6,3%.After some observations, which indicated a low prevalence of Salmonella infections in the area of Araraquara, S. Paulo, an extended schedule of enrichment and isolation media were applied for investigating the presence of Salmonella organisms in 47 fecal specimens from infantile gastroenterites, 51 stool specimens from animais and 50 food samples of animal origin. Salmonella isolates were obtained in 6,3% of the g astro enter itis cases; 5,8% from animal stool cultures and 8,0% from food stuffs. In the last case only pork liver gave positive results, with no isolations being achieved from beef Products. S. anatum, S. derby and S. daytona were found in descendiny order of frequency. As related to the gastroenteritis cases Shigella organisms were twice as frequent than the Salmonella.

Fate of Salmonella enterica and Enterohemorrhagic Escherichia coli Cells Artificially Internalized into Vegetable Seeds during Germination.

Science.gov (United States)

Liu, Da; Cui, Yue; Walcott, Ronald; Chen, Jinru

2018-01-01

Vegetable seeds contaminated with bacterial pathogens have been linked to fresh-produce-associated outbreaks of gastrointestinal infections. This study was undertaken to observe the physiological behavior of Salmonella enterica and enterohemorrhagic Escherichia coli (EHEC) cells artificially internalized into vegetable seeds during the germination process. Surface-decontaminated seeds of alfalfa, fenugreek, lettuce, and tomato were vacuum-infiltrated with four individual strains of Salmonella or EHEC. Contaminated seeds were germinated at 25°C for 9 days, and different sprout/seedling tissues were microbiologically analyzed every other day. The internalization of Salmonella and EHEC cells into vegetable seeds was confirmed by the absence of pathogens in seed-rinsing water and the presence of pathogens in seed homogenates after postinternalization seed surface decontamination. Results show that 317 (62%) and 343 (67%) of the 512 collected sprout/seedling tissue samples were positive for Salmonella and EHEC, respectively. The average Salmonella populations were significantly larger ( P seed coat tissues, followed by the root tissues, but the mean EHEC populations from all sampled tissue sections were statistically similar, except in pregerminated seeds. Three Salmonella and two EHEC strains had significantly larger cell populations on sprout/seedling tissues than other strains used in the study. Salmonella and EHEC populations from fenugreek and alfalfa tissues were significantly larger than those from tomato and lettuce tissues. The study showed the fate of internalized human pathogens on germinating vegetable seeds and sprout/seedling tissues and emphasized the importance of using pathogen-free seeds for sprout production. IMPORTANCE The internalization of microorganisms into vegetable seeds could occur naturally and represents a possible pathway of vegetable seed contamination by human pathogens. The present study investigated the ability of two important

Antimicrobial activity of some of the south-Indian spices against serotypes of Escherichia coli, Salmonella, Listeria monocytogenes and Aeromonas hydrophila Atividade antimicrobiana de condimentos do sul da India sobre Escherichia coli, Salmonella, Listeria monocytogenes e Aeromonas hydrophila

Directory of Open Access Journals (Sweden)

M.N. Indu

2006-06-01

Full Text Available Antibacterial activity of extracts of Allium sativum (garlic, Myristica fragrans (nutmeg, Zingiber officinale (ginger, Allium cepa (onion and Piper nigrum (pepper has been evaluated against 20 different serogroups of Escherichia coli, 8 serotypes of Salmonella, Listeria monocytogenes and Aeromonas hydrophila. Garlic extract showed excellent antibacterial activity against all the test organisms, except L. monocytogenes. Nutmeg showed good anti-listerial activity, although activity against E. coli and Salmonella were serotype dependent. Both garlic and nutmeg extracts were effective against A. hydrophila. Extracts of ginger showed inhibitory activity against two serogroups of E. coli: as O8 (enterotoxigenic E. coli and O88 only. Extracts of onion and pepper did not show any antibacterial activity against the test organisms.Avaliou-se a atividade antimicrobiana de extratos de alho (Allium sativum, noz-moscada (Mysritica frangrans, gengibre (Zingiber officinale cebola (Allium cepa e pimenta do reino (Piper nigrum sobre 20 sorotipos de Escherichia coli, 8 sorotipos de Salmonella, Listeria monocytogenes e Aeromonas hydrophila. O alho apresentou atividade antimicrobiana excelente sobre todos os microrganismos testados, excepto L. monocytogenes. A noz-moscada apresentou boa atividade antilisteria, emboara atividade sobre E. coli e Salmonella tenha sido sorotipo-dependente. Tanto alho como noz-moscada foram eficientes contra A. hydrophila. O extrato de gengibre apresentou atividade inibitória sobre dois sorotipos de E. coli: 08 (enterotoxigenico e 088. Os extratos de cebola e pimenta do reino não apresentaram nenhuma atividade contra os microrganismos testados.

Study of Shigella Vaccines in Man.

Science.gov (United States)

1974-08-15

and invasive Escherichia coli diarrheal disease, acute non-bacterial gastro- enteritis of adults ( parvovirus ) and choler;-j-For all the above-mentioned...with: 1) Enterotoxigenic E. coli. 2) Invasive E. coli. 3) Parvoviruses . 4) Giardia intestinalis. C. To investigate both the non-specific and specific... attenuated .trains of shigell:i for kise as oral vaccines. Presently we are involved ir, evalua. ion of the ’third gene: ioi " of oral shigella

Salmonella, Escherichia coli and Enterobacteriaceae in the peanut supply chain: From farm to table.

Science.gov (United States)

Nascimento, M S; Carminati, J A; Silva, I C R N; Silva, D L; Bernardi, A O; Copetti, M V

2018-03-01

Due to recent foodborne outbreaks, peanuts have been considered a potential risk for Salmonella transmission. For this reason, the aim of this study was to determine the prevalence and contamination load of Salmonella, Escherichia coli and Enterobacteriaceae throughout the peanut supply chain in Brazil. Samples of peanuts and peanut-containing processed products from post-harvest (n=129), secondary processing (n=185) and retail market (n=100) were analyzed. The results showed high Enterobacteriaceae counts in the post-harvest samples. At the end of the secondary processing, 16% of the samples remained contaminated by this group of microorganisms. Six peanut samples from primary production and one sample of peanut butter were tested positive for E. coli while Salmonella was detected in nine samples (2.2%): six from post-harvest, two from the initial stage of the secondary processing and one from retail. The Salmonella counts ranged between 0.004 and 0.092MPN/g and five serotypes were identified (Muenster, Miami, Javiana, Oranienburg, Glostrup). The results demonstrated a high prevalence of Enterobacteriaceae and low prevalence of E. coli throughout the peanut supply chain. Furthermore, it was verified that peanuts may become contaminated by Salmonella during different stages of the supply chain, especially at post-harvest. Copyright © 2017 Elsevier Ltd. All rights reserved.

lac repressor is an antivirulence factor of Salmonella enterica: its role in the evolution of virulence in Salmonella.

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Sandeepa M Eswarappa

Full Text Available The genus Salmonella includes many pathogens of great medical and veterinary importance. Bacteria belonging to this genus are very closely related to those belonging to the genus Escherichia. lacZYA operon and lacI are present in Escherichia coli, but not in Salmonella enterica. It has been proposed that Salmonella has lost lacZYA operon and lacI during evolution. In this study, we have investigated the physiological and evolutionary significance of the absence of lacI in Salmonella enterica. Using murine model of typhoid fever, we show that the expression of LacI causes a remarkable reduction in the virulence of Salmonella enterica. LacI also suppresses the ability of Salmonella enterica to proliferate inside murine macrophages. Microarray analysis revealed that LacI interferes with the expression of virulence genes of Salmonella pathogenicity island 2. This effect was confirmed by RT-PCR and Western blot analysis. Interestingly, we found that SBG0326 of Salmonella bongori is homologous to lacI of Escherichia coli. Salmonella bongori is the only other species of the genus Salmonella and it lacks the virulence genes of Salmonella pathogenicity island 2. Overall, our results demonstrate that LacI is an antivirulence factor of Salmonella enterica and suggest that absence of lacI has facilitated the acquisition of virulence genes of Salmonella pathogenicity island 2 in Salmonella enterica making it a successful systemic pathogen.

Bacterial pathogens associated with bloody diarrhea in Uruguayan children Patógenos bacterianos asociados a diarrea con sangre en niños uruguayos

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M. I. Mota

2010-06-01

Full Text Available Diarrheal disease continues to be a serious health problem, especially in developing countries. Bloody diarrhea represents approximately 20-30% of all cases and has higher morbidity and mortality. Treatment with antibiotics is beneficial in cases of Shigella, Campylobacter, Yersinia and Salmonella infection, principally in those children with a higher risk of invasive disease. The aims of this study were to detect the bacterial agents associated with bloody diarrhea in children and to determine their antimicrobial susceptibility patterns. Between June 2001 and January 2008, 249 children with bloody diarrhea were studied. Shigella and Shiga toxin-producing Escherichia coli (STEC were recovered from 48 (19.3% and 3 (1.2% of the total of cases, respectively. In 49 out of 249 children, in whom other enteropathogens were investigated, we recovered Campylobacter jejuni from 7 children (14.3%, Salmonella spp. from 2 (4.1% and Aeromonas spp. from 1 (2% in addition to Shigella from 7 children (14.3%. Thirty-four (70% Shigella isolates showed resistance to ampicillin and 13 (27% to trimethoprim-sulfamethoxazole. All Shigella isolates were susceptible to nalidixic acid, ciprofloxacin and ceftriaxone. Salmonella and STEC isolates were susceptible to all antibiotics assayed. Thus, the use of trimethoprim-sulfamethoxazole or ampicillin would not be appropriate for the empirical treatment of Shigella - associated diarrhea.La enfermedad diarreica es un problema de salud importante, especialmente en los países en desarrollo. La diarrea sanguinolenta representa el 20-30% del total de casos y determina una mayor morbimortalidad. La antibioticoterapia es beneficiosa en casos de infección por Shigella, Campylobacter, Yersinia y Salmonella, principalmente en niños con riesgo elevado de enfermedad invasora. Los objetivos del presente trabajo fueron conocer las bacterias asociadas a diarrea sanguinolenta y determinar su patrón de sensibilidad a los antimicrobianos

Electrochemical genosensing of Salmonella, Listeria and Escherichia coli on silica magnetic particles

Energy Technology Data Exchange (ETDEWEB)

Liébana, Susana; Brandão, Delfina [Grup de Sensors i Biosensors, Departament de Química, Universitat Autònoma de Barcelona, 08193, Cerdanyola del Vallès (Bellaterra) (Spain); Cortés, Pilar; Campoy, Susana [Departament de Genètica i de Microbiologia, Universitat Autònoma de Barcelona, 08193, Cerdanyola del Vallès (Bellaterra) (Spain); Alegret, Salvador [Grup de Sensors i Biosensors, Departament de Química, Universitat Autònoma de Barcelona, 08193, Cerdanyola del Vallès (Bellaterra) (Spain); Pividori, María Isabel, E-mail: Isabel.Pividori@uab.cat [Grup de Sensors i Biosensors, Departament de Química, Universitat Autònoma de Barcelona, 08193, Cerdanyola del Vallès (Bellaterra) (Spain)

2016-01-21

A magneto-genosensing approach for the detection of the three most common pathogenic bacteria in food safety, such as Salmonella, Listeria and Escherichia coli is presented. The methodology is based on the detection of the tagged amplified DNA obtained by single-tagging PCR with a set of specific primers for each pathogen, followed by electrochemical magneto-genosensing on silica magnetic particles. A set of primers were selected for the amplification of the invA (278 bp), prfA (217 bp) and eaeA (151 bp) being one of the primers for each set tagged with fluorescein, biotin and digoxigenin coding for Salmonella enterica, Listeria monocytogenes and E. coli, respectively. The single-tagged amplicons were then immobilized on silica MPs based on the nucleic acid-binding properties of silica particles in the presence of the chaotropic agent as guanidinium thiocyanate. The assessment of the silica MPs as a platform for electrochemical magneto-genosensing is described, including the main parameters to selectively attach longer dsDNA fragments instead of shorter ssDNA primers based on their negative charge density of the sugar-phosphate backbone. This approach resulted to be a promising detection tool with sensing features of rapidity and sensitivity very suitable to be implemented on DNA biosensors and microfluidic platforms. - Highlights: • Silica magnetic particles were used for the first time as carrier in electrochemical magneto-genosensing of single-tagged amplicons. • They demonstrated to be a robust platform for the electrochemical detection of PCR products. • Differential adsorption properties for longer dsDNA amplicon incorporating the tagging primers over shorter ssDNA tagged primers were observed due to the negative charge density. • Electrochemical magneto-genosensing of Salmonella enterica, Listeria monocytogenes and Escherichia coli was successfully performed.

Electrochemical genosensing of Salmonella, Listeria and Escherichia coli on silica magnetic particles

International Nuclear Information System (INIS)

Liébana, Susana; Brandão, Delfina; Cortés, Pilar; Campoy, Susana; Alegret, Salvador; Pividori, María Isabel

2016-01-01

A magneto-genosensing approach for the detection of the three most common pathogenic bacteria in food safety, such as Salmonella, Listeria and Escherichia coli is presented. The methodology is based on the detection of the tagged amplified DNA obtained by single-tagging PCR with a set of specific primers for each pathogen, followed by electrochemical magneto-genosensing on silica magnetic particles. A set of primers were selected for the amplification of the invA (278 bp), prfA (217 bp) and eaeA (151 bp) being one of the primers for each set tagged with fluorescein, biotin and digoxigenin coding for Salmonella enterica, Listeria monocytogenes and E. coli, respectively. The single-tagged amplicons were then immobilized on silica MPs based on the nucleic acid-binding properties of silica particles in the presence of the chaotropic agent as guanidinium thiocyanate. The assessment of the silica MPs as a platform for electrochemical magneto-genosensing is described, including the main parameters to selectively attach longer dsDNA fragments instead of shorter ssDNA primers based on their negative charge density of the sugar-phosphate backbone. This approach resulted to be a promising detection tool with sensing features of rapidity and sensitivity very suitable to be implemented on DNA biosensors and microfluidic platforms. - Highlights: • Silica magnetic particles were used for the first time as carrier in electrochemical magneto-genosensing of single-tagged amplicons. • They demonstrated to be a robust platform for the electrochemical detection of PCR products. • Differential adsorption properties for longer dsDNA amplicon incorporating the tagging primers over shorter ssDNA tagged primers were observed due to the negative charge density. • Electrochemical magneto-genosensing of Salmonella enterica, Listeria monocytogenes and Escherichia coli was successfully performed.

Recovery of Cephalosporin Resistant Escherichia coli and Salmonella from Pork, Beef and Chicken Marketed in Nova Scotia

Directory of Open Access Journals (Sweden)

Kevin R Forward

2004-01-01

Full Text Available BACKGROUND: Antimicrobial use in farm animals is a potentially important contributor to the emergence of antimicrobial resistance. Resistant Salmonella may lead to serious human infections and resistant Escherichia coli may transfer plasmid-encoded resistance genes to other pathogens.

Gamma radiation in the reduction of Salmonella spp. inoculated on minimally processed watercress (Nasturtium officinalis)

Energy Technology Data Exchange (ETDEWEB)

Martins, C.G.; Behrens, J.H.; Destro, M.T.; Franco, B.D.G.M.; Vizeu, D.M.; Hutzler, B.; Landgraf, M. E-mail: landgraf@usp.br

2004-10-01

Consumer attitudes towards foods have changed in the last two decades increasing requirements for freshlike products. Consequently, less extreme treatments or additives are being required. Minimally processed foods have freshlike characteristics and satisfy this new consumer demand. Besides freshness, the minimally processing also provide convenience required by the market. Salad vegetables can be source of pathogen such as Salmonella, Escherichia coli O157:H7, Shigella spp. The minimal processing does not reduce the levels of pathogenic microorganisms to safe levels. Therefore, this study was carried out in order to improve the microbiological safety and the shelf-life of minimally processed vegetables using gamma radiation. Minimally processed watercress inoculated with a cocktail of Salmonella spp was exposed to 0.0, 0.2, 0.5, 0.7, 1.0, 1.2 and 1.5 kGy. Irradiated samples were diluted 1:10 in saline peptone water and plated onto tryptic soy agar that were incubated at 37 deg. C/24 h. D{sub 10} values for Salmonella spp. inoculated in watercress varied from 0.29 to 0.43 kGy. Therefore, a dose of 1.7 kGy will reduce Salmonella population in watercress by 4 log{sub 10}. The shelf-life was increased by 1 1/2 day when the product was exposed to 1 kGy.

Investigating the decay rates of Escherichia coli relative to Vibrio parahemolyticus and Salmonella Typhi in tropical coastal waters.

Science.gov (United States)

Lee, Choon Weng; Ng, Angie Yee Fang; Bong, Chui Wei; Narayanan, Kumaran; Sim, Edmund Ui Hang; Ng, Ching Ching

2011-02-01

Using the size fractionation method, we measured the decay rates of Escherichia coli, Salmonella Typhi and Vibrio parahaemolyticus in the coastal waters of Peninsular Malaysia. The size fractions were total or unfiltered, 0.7 μm) than in the smaller fraction (Vibrio grew well in seawater. There was usually an increase in Vibrio after one day incubation. Our results confirmed that decay or loss rates of E. coli did not match that of Vibrio, and also did not correlate with Salmonella decay rates. However E. coli showed persistence where its decay rates were generally lower than Salmonella. © 2010 Elsevier Ltd. All rights reserved.

Long-term variations in abundance and distribution of sewage pollution indicator and human pathogenic bacteria along the central west coast of India

Digital Repository Service at National Institute of Oceanography (India)

Rodrigues, V.; Ramaiah, N.; Kakti, S.; Samant, D.

- FC and Escherichia coli - EC) and potential pathogens (Vibrio cholerae - VC, Shigella - SH, and Salmonella spp. - SA). In both Mandovi and Zuari estuaries, where fishing and tourist-related activities are sizable and long-term data collection...

Inactivation of Escherichia coli O157:H7 and Salmonella on fresh herbs by plant essential oils

Science.gov (United States)

Consumer awareness of fresh herbs and its demand has increased in recent years due to health benefits and distinct aroma in prepared food. There are specific markets for local growers, especially for organically grown herbs. Shiga-toxigenic Escherichia coli and Salmonella spp. have been detected and...

Rapid and reliable discrimination between Shigella species and Escherichia coli using MALDI-TOF mass spectrometry

NARCIS (Netherlands)

Paauw, A.; Jonker, D.; Roeselers, G.; Heng, J.M.E.; Mars-Groenendijk, R.H.; Trip, H.; Molhoek, E.M.; Jansen, H.J.; Plas, J. van der; Jong, A.L. de; Majchrzykiewicz-Koehorst, J.A.; Speksnijder, A.G.C.L.

2015-01-01

E. coli-. Shigella species are a cryptic group of bacteria in which the Shigella species are distributed within the phylogenetic tree of E. coli. The nomenclature is historically based and the discrimination of these genera developed as a result of the epidemiological need to identify the cause of

Polymerase chain reaction for detection of invasive Shigella flexneri in food.

Science.gov (United States)

Lampel, K A; Jagow, J A; Trucksess, M; Hill, W E

1990-06-01

The polymerase chain reaction (PCR) was used to amplify a 760-base-pair (bp) fragment with the 220-kbp invasive plasmids of enteroinvasive Escherichia coli, Shigella flexneri, Shigella dysenteriae, Shigella boydii, and Shigella sonnei as templates. This PCR product was easily detected by agarose gel electrophoresis. A 210-bp AccI-PstI fragment lying within the amplified region was used as a probe in Southern hybridization blots and showed that the PCR-generated product was derived from the invasive plasmid. The application of PCR as a rapid method to detect enteroinvasive bacteria in foods was tested by inoculating lettuce with 10(4) S. flexneri cells per g in shigella broth base. Plasmid DNA was isolated from cultures of inoculated and uninoculated lettuce in broth after 0, 4, and 24 h of incubation. With the PCR, the 760-bp fragment was generated only from lettuce inoculated with S. flexneri, as shown by gel electrophoresis and confirmed both by Southern blotting and by nucleotide sequencing of the amplified region. Because the isolation of plasmid DNA, the performance of PCR, and gel electrophoresis all can be completed in 6 to 7 h, invasive enteric bacteria can be detected in less than 1 day.

Surveys monitoring the sea and beaches in the vicinity of Durban, South Africa: a case study

CSIR Research Space (South Africa)

Rathbone, PA

1998-01-01

Full Text Available until the present day. A classification system including Escherichia coli I, helminthic parasite ova, pathogenic staphylococci, salmonellae, shigellae, and salinity was used. This system, functioning as an audit, has proved useful in detecting problem...

[Active etiological surveillance for foodborne diseases in Guangdong province, 2013-2014

NARCIS (Netherlands)

Ke, B. X.; He, D. M.; Tan, H. L.; Zeng, H. H.; Yang, T.; Li, B. S.; Liang, Y. H.; Lu, L. L.; Liang, J. H.; Huang, Q.; Ke, C. W.

2016-01-01

Objective: To study the infection status, serotypes, drug resistance and molecular characteristics of Salmonella, Shigella, Vibrio parahemolyticus, enterotoxigenic Escherichia (E.) coli (ETEC), pathogenic E. coli (EPEC), Shiga Toxin producing E. coli (STEC) and Enteroinvasive E. coli (EIEC)

Computational determination of the effects of virulent Escherichia coli and salmonella bacteriophages on human gut.

Science.gov (United States)

Mostafa, Marwa Mostafa; Nassef, Mohammad; Badr, Amr

2016-10-01

Salmonella and Escherichia coli are different types of bacteria that cause food poisoning in humans. In the elderly, infants and people with chronic conditions, it is very dangerous if Salmonella or E. coli gets into the bloodstream and then they must be treated by phage therapy. Treating Salmonella and E. coli by phage therapy affects the gut flora. This research paper presents a system for detecting the effects of virulent E. coli and Salmonella bacteriophages on human gut. A method based on Domain-Domain Interactions (DDIs) model is implemented in the proposed system to determine the interactions between the proteins of human gut bacteria and the proteins of bacteriophages that infect virulent E. coli and Salmonella. The system helps gastroenterologists to realize the effect of injecting bacteriophages that infect virulent E. coli and Salmonella on the human gut. By testing the system over Enterobacteria phage 933W, Enterobacteria phage VT2-Sa and Enterobacteria phage P22, it resulted in four interactions between the proteins of the bacteriophages that infect E. coli O157:H7, E. coli O104:H4 and Salmonella typhimurium and the proteins of human gut bacterium strains. Several effects were detected such as: antibacterial activity against a number of bacterial species in human gut, regulation of cellular differentiation and organogenesis during gut, lung, and heart development, ammonia assimilation in bacteria, yeasts, and plants, energizing defense system and its function in the detoxification of lipopolysaccharide, and in the prevention of bacterial translocation in human gut. Copyright © 2016 Elsevier Ireland Ltd. All rights reserved.

Antibacterial activity of some commonly used food commodities against escherichia coli, salmonella typhi and staphylococcus aureus

International Nuclear Information System (INIS)

Siddiqui, A.; Ansari, A.

2009-01-01

The activity of commonly used spices and salt, sugar and pickles against Escherichia coli, Salmonella typhi and staphlococcus aureus was tested. The antibacterial activity was found to be in descending order like coriander>pickles>salt and sugar>clove>black pepper>red chilli against S. typhi and garlic>clove>onion>ginger against S. aureus. (author)

Rapid detection of Shigella and enteroinvasive Escherichia coli in produce enrichments by a conventional multiplex PCR assay.

Science.gov (United States)

Binet, Rachel; Deer, Deanne M; Uhlfelder, Samantha J

2014-06-01

Faster detection of contaminated foods can prevent adulterated foods from being consumed and minimize the risk of an outbreak of foodborne illness. A sensitive molecular detection method is especially important for Shigella because ingestion of as few as 10 of these bacterial pathogens can cause disease. The objectives of this study were to compare the ability of four DNA extraction methods to detect Shigella in six types of produce, post-enrichment, and to evaluate a new and rapid conventional multiplex assay that targets the Shigella ipaH, virB and mxiC virulence genes. This assay can detect less than two Shigella cells in pure culture, even when the pathogen is mixed with background microflora, and it can also differentiate natural Shigella strains from a control strain and eliminate false positive results due to accidental laboratory contamination. The four DNA extraction methods (boiling, PrepMan Ultra [Applied Biosystems], InstaGene Matrix [Bio-Rad], DNeasy Tissue kit [Qiagen]) detected 1.6 × 10(3)Shigella CFU/ml post-enrichment, requiring ∼18 doublings to one cell in 25 g of produce pre-enrichment. Lower sensitivity was obtained, depending on produce type and extraction method. The InstaGene Matrix was the most consistent and sensitive and the multiplex assay accurately detected Shigella in less than 90 min, outperforming, to the best of our knowledge, molecular assays currently in place for this pathogen. Published by Elsevier Ltd.

Antimicrobial-resistant patterns of Escherichia coli and Salmonella strains in the aquatic Lebanese environments

International Nuclear Information System (INIS)

Harakeh, Steve; Yassine, Hadi; El-Fadel, Mutasem

2006-01-01

This study is the first to be conducted in Lebanon on the isolation and molecular characterization and the antimicrobial resistance profile of environmental pathogenic bacterial strains. Fifty-seven samples of seawater, sediment, crab, and fresh water were collected during the spring and summer seasons of 2003. The isolation of Escherichia coli and Salmonella using appropriate selective media revealed that 94.7% of the tested samples were contaminated with one or both of the tested bacteria. The polymerase chain reaction (PCR) was then used to identify the species of both bacteria using various sets of primers. Many pathogenic E. coli isolates were detected by PCR out of which two were identified as O157:H7 E. coli. Similarly, the species of many of the Salmonella isolates was molecularly identified. The confirmed isolates of Salmonella and E. coli were then tested using the disk diffusion method for their susceptibility to four different antimicrobials revealing high rates of antimicrobial resistance. - First report of antibiotic resistance in bacteria in the environment in Lebanon

[Occurrence of Salmonella spp. and shigatoxin-producing escherichia coli (STEC) in horse faeces and horse meat products].

Science.gov (United States)

Pichner, Rohtraud; Sander, Andrea; Steinrück, Hartmut; Gareis, Manfred

2005-01-01

In order to assess the relevance of horses as a possible reservoir of Salmonella and Shigatoxin-producing Escherichia coli (STEC), 400 samples of horse faeces and 100 samples of horse meat products were examined by PCR-screening methods. Salmonella enterica was not found in any of the samples. One faeces-sample and one horse meat product were proved to be STEC positive. The STEC-strain from faecal origin belonged to the serotype 0113:H21 and had the stx 2c gene and the enterohemolysin gene. The STEC-strain isolated from a horse meat product had the serotype O87:H16 and the stx 2d gene. The results indicate a very low risk for human to get a Salmonella- or EHEC- infection from horses in Germany.

Cellular damage of Escherichia coli 0157:H7 and Salmonella spp. in apple juice treated with high hydrostatic pressure and thermal death time disks

Science.gov (United States)

Differences in membrane damage including leakage of intracellular UV-materials and loss of viability of Salmonella spp. and Escherichia coli O157:H7 bacteria in apple juice, pH 3.1 following thermal-death-time (TDT) disk and high hydrostatic pressure (HHP) treatments were investigated. Salmonella an...

Failure of Syndrome-Based Diarrhea Management Guidelines to Detect Shigella Infections in Kenyan Children.

Science.gov (United States)

Pavlinac, P B; Denno, D M; John-Stewart, G C; Onchiri, F M; Naulikha, J M; Odundo, E A; Hulseberg, C E; Singa, B O; Manhart, L E; Walson, J L

2016-12-01

Shigella is a leading cause of childhood diarrhea mortality in sub-Saharan Africa. Current World Health Organization guidelines recommend antibiotics for children in non cholera-endemic areas only in the presence of dysentery, a proxy for suspected Shigella infection. To assess the sensitivity and specificity of the syndromic diagnosis of Shigella-associated diarrhea, we enrolled children aged 6 months to 5 years presenting to 1 of 3 Western Kenya hospitals between November 2011 and July 2014 with acute diarrhea. Stool samples were tested using standard methods for bacterial culture and multiplex polymerase chain reaction for pathogenic Escherichia coli. Stepwise multivariable logit models identified factors to increase the sensitivity of syndromic diagnosis. Among 1360 enrolled children, median age was 21 months (interquartile range, 11-37), 3.4% were infected with human immunodeficiency virus, and 16.5% were stunted (height-for-age z-score less than -2). Shigella was identified in 63 children (4.6%), with the most common species being Shigella sonnei (53.8%) and Shigella flexneri (40.4%). Dysentery correctly classified 7 of 63 Shigella cases (sensitivity, 11.1%). Seventy-eight of 1297 children without Shigella had dysentery (specificity, 94.0%). The combination of fecal mucous, age over 23 months, and absence of excessive vomiting identified more children with Shigella-infection (sensitivity, 39.7%) but also indicated antibiotics in more children without microbiologically confirmed Shigella (specificity, 82.7%). Reliance on dysentery as a proxy for Shigella results in the majority of Shigella-infected children not being identified for antibiotics. Field-ready rapid diagnostics or updated evidence-based algorithms are urgently needed to identify children with diarrhea most likely to benefit from antibiotic therapy. © The Author 2015. Published by Oxford University Press on behalf of the Pediatric Infectious Diseases Society. All rights reserved. For Permissions

Characterisation of Shigella species isolated from river catchments in the North West province of South Africa

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Constance Wose Kinge

2010-10-01

Full Text Available The occurrence and distribution of Shigella species in water from the five river catchments in the North West province of South Africa were investigated. Shigella is a Gram-negative, non-motile, facultative anaerobic bacillus that causes shigellosis, an important cause of morbidity and mortality in high-risk populations (such as children, the elderly and immuno-compromised individuals that depend on river water. A total of 54 water samples collected in winter (April 2007 to July 2007 and summer (December 2007 to March 2008 were cultured on Salmonella-Shigella agar by the spread-plate method. Suspected Shigella isolates obtained were characterised by primary biochemical (Triple Sugar Iron agar and agglutination and molecular (polymerase chain reactions, PCR tests. Amplification of the invasion plasmid gene (ipaH by PCR was done to confirm the presence of Shigella spp. in water. In total, 214 Shigella boydii, 15 Shigella dysenteriae, 11 Shigella flexneri and 2 Shigella sonnei were confirmed by serotyping in both winter and summer samples. The ipaH gene (606 bp was present in 176 and 49 of the winter and summer isolates, respectively. The presence of Shigella spp. in water was confirmed with over 90% specificity. The need for more effective management of these river catchments and the provision of potable water and sanitation facilities is needed to minimise the occurrence and transmission of water-borne diseases caused by these and other pathogenic bacteria.

New strategies for genetic engineering Pseudomonas syringae using recombination

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Here we report that DNA oligonucleotides (oligos) introduced directly into bacteria by electroporation can recombine with the bacterial chromosome. This phenomenon was identified in Pseudomonas syringae and we subsequently found that Escherichia coli, Salmonella typhimurium and Shigella flexneri are...

Expression of bacterial virulence factors and cytokines during in vitro macrophage infection by enteroinvasive Escherichia coli and Shigella flexneri: a comparative study

Directory of Open Access Journals (Sweden)

Silvia Y Bando

2010-09-01

Full Text Available Enteroinvasive Escherichia coli (EIEC and Shigellaspp cause bacillary dysentery in humans by invading and multiplying within epithelial cells of the colonic mucosa. Although EIEC and Shigellashare many genetic and biochemical similarities, the illness caused by Shigellais more severe. Thus, genomic and structure-function molecular studies on the biological interactions of these invasive enterobacteria with eukaryotic cells have focused on Shigella rather than EIEC. Here we comparatively studied the interactions of EIEC and of Shigella flexneriwith cultured J774 macrophage-like cells. We evaluated several phenotypes: (i bacterial escape from macrophages after phagocytosis, (ii macrophage death induced by EIEC and S. flexneri, (iii macrophage cytokine expression in response to infection and (iv expression of plasmidial (pINV virulence genes. The results showed thatS. flexneri caused macrophage killing earlier and more intensely than EIEC. Both pathogens induced significant macrophage production of TNF, IL-1 and IL-10 after 7 h of infection. Transcription levels of the gene invasion plasmid antigen-C were lower in EIEC than in S. flexneri throughout the course of the infection; this could explain the diminished virulence of EIEC compared to S. flexneri.

Prevalence and Antibiogram Profile of Salmonellae in Intensively ...

African Journals Online (AJOL)

Prof. Ogunji

Authors are very thankful to Dr. Simon Le Hello,. Co-directeur Centre National de Référence E. coli/Shigella/Salmonella Institut Pasteur, for serotyping of the isolates, and also to Dr. Oladepo Dauda of the National Institute for. Pharmaceutical Research and Development. (NIPRD), Abuja for providing the reference.

Role of the SRRz/Rz1 lambdoid lysis cassette in the pathoadaptive evolution of Shigella.

Science.gov (United States)

Leuzzi, Adriano; Grossi, Milena; Di Martino, Maria Letizia; Pasqua, Martina; Micheli, Gioacchino; Colonna, Bianca; Prosseda, Gianni

2017-06-01

Shigella, the etiological agent of bacillary dysentery (shigellosis), is a highly adapted human pathogen. It evolved from an innocuous ancestor resembling the Escherichia coli strain by gain and loss of genes and functions. While the gain process concerns the acquisition of the genetic determinants of virulence, the loss is related to the adaptation of the genome to the new pathogenic status and occurs by pathoadaptive mutation of antivirulence genes. In this study, we highlight that the SRRz/Rz 1 lambdoid lysis cassette, even though stably adopted in E. coli K12 by virtue of its beneficial effect on cell physiology, has undergone a significant decay in Shigella. Moreover, we show the antivirulence nature of the SRRz/Rz 1 lysis cassette in Shigella. In fact, by restoring the SRRz/Rz 1 expression in this pathogen, we observe an increased release of peptidoglycan fragments, causing an unbalance in the fine control exerted by Shigella on host innate immunity and a mitigation of its virulence. This strongly affects the virulence of Shigella and allows to consider the loss of SRRz/Rz 1 lysis cassette as another pathoadaptive event in the life of Shigella. Copyright © 2017 Elsevier GmbH. All rights reserved.

Effect of the oyster contaminated in Salmonella typhimurium and Escherichia coli

International Nuclear Information System (INIS)

Brena V, M.

1992-08-01

In this work the effect of the oyster contaminated in the species of bacteria better studied by its genetic composition these are the Salmonella typhimurium and the Escherichia coli and that its have been starting point for later radiobiological studies in superior organisms. Of this its have arisen two general lines. The research about the genotoxic effect of substances or mixtures in bacteria with the collaboration of the groups of Drosophila and X-ray Fluorescence analysis and on the other hand the study of the low doses of radiation also in bacteria. It is also treated the topic about the genetic effect of aromatic hydrocarbons in different biological systems. (Author)

2262-IJBCS-Article-Charles Fokunang

African Journals Online (AJOL)

hp

qualitative phytochemical screening and the antimicrobial activities on Shigella dysenteriae A2, Salmonella typhi, Escherichia coli, Enterococcus ... screening was performed using colorimetric methods. The antimicrobial activity was carried out ... plants through high throughput modern technics, and looking for new leads to.

Assessment of yeast biofilter in the treatment of domestic wastewater ...

African Journals Online (AJOL)

The bacteria Staphylococcus aureus, Escherichia coli, Proteus mirabilis, Serratia spp, Pseudomanas spp, Shigella spp, Salmonella spp, Bacillus cereus and the fungi Aspergillus niger, Aspergillus fumigatus, Penicillum spp, Aspergillus flavus were completely eliminated after the biofiltration process. This shows the health ...

Prevalence of intestinal parasites, salmonella and shigella among apparently health food handlers of Addis Ababa University student's cafeteria, Addis Ababa, Ethiopia.

Science.gov (United States)

Aklilu, Addis; Kahase, Daniel; Dessalegn, Mekonnen; Tarekegn, Negatu; Gebremichael, Saba; Zenebe, Seyfe; Desta, Kassu; Mulugeta, Gebru; Mamuye, Yeshiwodim; Mama, Mohammedaman

2015-01-24

Food contamination may occur at any point during its journey through production, processing, distribution, and preparation. The risk of food getting contaminated depends largely on the health status of the food handlers, their personal hygiene, knowledge and practice of food hygiene. Food borne diseases are a public health problem in developed and developing countries like Ethiopia. A cross sectional study was conducted among food handlers in Addis Ababa student's cafeteria from January to May 2013. Structured questionnaire was used to collect socio demographic data and associated risk factors. Stool specimens were examined for bacteria and intestinal parasites following standard procedures. Biochemical tests were done to identify the species of bacterial isolates. Sensitivity testing was done using Kirby- Baur disk diffusion method. A total of 172 food handlers were enrolled in the study. The majority of study participants were females 134 (77.9%). About 78 (45.3%) of food handlers were found to be positive for different intestinal parasites with the most abundant parasite of Entameoba histolytica/dispar 68 (70.8%) followed by Giardia lamblia 18 (18.8%), Taenia species 5 (5.2%), Ascaris lumbricoides 2 (2.1%), hookworm 2 (2.1%) and Trichuris trichiura 1 (1.1%). Stool cultures revealed 3.5% of Salmonella isolates (Sero-grouping on Salmonella isolate was not done), while Shigella species was not isolated from any of the stool samples obtained from Food handlers. All isolates of Salmonella were sensitive to ciprofloxacin, amikacin and gentamicin but resistant to ampicillin, clindamycin, and erythromycin. The present study revealed a high prevalence of intestinal parasite in asymptomatic (apparently health) food handlers. Such infected food handlers can contaminate food, drinks and could serve as source of infection to consumers via food chain.

Development of recombinant vaccine candidate molecule against Shigella infection.

Science.gov (United States)

Chitradevi, S T S; Kaur, G; Sivaramakrishna, U; Singh, D; Bansal, A

2016-10-17

Shigellosis is an acute bacillary diarrheal disease caused by the gram negative bacillus Shigella. The existence of multiple Shigella serotypes and their growing resistance to antibiotics stress the urgent need for the development of vaccine that is protective across all serotypes. Shigella's IpaB antigen is involved in translocon pore formation, promotes bacterial invasion and induces apoptosis in macrophages. S. Typhi GroEL (Hsp 60) is the immunodominant antigen inducing both arms of immunity and has been explored as adjuvant in this study. The present study evaluates the immunogenicity and protective efficacy of recombinant IpaB domain-GroEL fusion protein in mice against lethal Shigella infection. The IpaB domain and GroEL genes were fused using overlap extension PCR and cloned in pRSETA expression vector. Fused gene was expressed in Escherichia coli BL-21 cells and the resulting 90 KDa fusion protein was purified by affinity chromatography. Intranasal (i.n.) immunization of mice with fusion protein increased the IgG and IgA antibody titers as compared to the group immunized with IpaB and GroEL and control PBS immunized group. Also IgG1 and IgG2a antibodies induced in fusion protein immunized mice were higher than co-immunized group. Significant increase in lymphocyte proliferation and cytokine levels (IFN-γ, IL-4 and IL-10), indicates induction of both Th1 and Th2 immune responses in both immunized groups. Immunization with fusion protein protected 90-95% of mice whereas 80-85% survivability was observed in co-immunized group against lethal challenge with S. flexneri, S. boydii and S. sonnei. Passive immunization conferred 60-70% protection in mice against all these Shigella species. Organ burden and histopathology studies also revealed significant decrease in lung infection as compared to the co-immunized group. Since IpaB is the conserved dominant molecule in all Shigella species, this study will lead to an ideal platform for the development of safe

[Active etiological surveillance for foodborne diseases in Guangdong province, 2013-2014].

Science.gov (United States)

Ke, B X; He, D M; Tan, H L; Zeng, H H; Yang, T; Li, B S; Liang, Y H; Lu, L L; Liang, J H; Huang, Q; Ke, C W

2016-10-10

Objective: To study the infection status, serotypes, drug resistance and molecular characteristics of Salmonella, Shigella, Vibrio parahemolyticus , enterotoxigenic Escherichia ( E. ) coli (ETEC), pathogenic E. coli (EPEC), Shiga Toxin producing E. coli (STEC) and Enteroinvasive E. coli (EIEC) collected from diarrhea patients in Guangdong. Methods: The strains of Salmonella, Shigella, V. parahemolyticus and 4 kinds of E. coli isolated from foodborne diseases surveillance during 2013-2014 were collected to conduct serotyping, drug resistance test and pulsed-field gel electrophoresis (PFGE). Results: A total of 3 372 stains of pathogens were isolated from 57 834 stool samples during 2013-2014, the overall positive rate was 5.83 % and the positive rate of Salmonella was highest, followed by that of V. parahemolyticus , 4 kinds of E. coli and Shigella . And 3 213 strains of Salmonella were divided into 143 serotypes. The most prevalent serotypes were Salmonella typhimurium , 4, 5, 12: i:-, Enteritidis , Stanley and Derby . Salmonella was sensitive to cephalosporin and fluoroquinolones, and showed significant differences in drug resistance rate among different serotypes. In top 10 common serotypes, S. enteritidis and S. derby were most resistant to cephalosporin and ciprofloxacin respectively. PFGE was performed for 2 289 strains of Salmonella , showing distribution diversity and significant fingerprint polymorphisms. The 85 strains of V. parahemolyticus were divided into 10 serotypes, O3∶K6 (61.18 % ) was the most common serotype, followed by O4∶K8. The results showed that the carrying rate of virulence genes tdh (81.18 % ) was high, while the carrying rate of trh was low (7.06 % ), and there were 10 strains carrying no the two kinds of virulence genes. The sensitive rate of V. parahemolyticus to imipenem, nalidixic acid, SMZ-TMP, chloramphenicol and tetracycline were more than 95 % . Thirteen strains of Shigella were detected, including 9 strains of Shigella

Radiosensitization of Escherichia coli and Salmonella typhi in presence of active compounds

International Nuclear Information System (INIS)

Lacroix, M.; Chiasson, F.; Borsa, J.; Ouattara, B.

2004-01-01

The radiosensitization of Escherichia coli and Salmonella typhi in ground beef was evaluated in the presence of 18 active compounds. Medium fat ground beef (23% fat) was inoculated with E. coli or S. typhi and each active compound was added separately at various concentrations. For E. coli, the most efficient compounds were trans-cinnamaldehyde, thymol and thyme. For S. typhi, the most efficient compounds was trans-cinnamaldehyde, carvacrol and thymol. The addition of tetrasodium pyrophosphate, carvacrol and ascorbic acid had no effect on the irradiation sensitivity of E. coli. For S. typhi, only ascorbic acid had no effect

Radiosensitization of Escherichia coli and Salmonella typhi in presence of active compounds

Energy Technology Data Exchange (ETDEWEB)

Lacroix, M. E-mail: monique.lacroix@inrs-iaf.uquebec.ca; Chiasson, F.; Borsa, J.; Ouattara, B

2004-10-01

The radiosensitization of Escherichia coli and Salmonella typhi in ground beef was evaluated in the presence of 18 active compounds. Medium fat ground beef (23% fat) was inoculated with E. coli or S. typhi and each active compound was added separately at various concentrations. For E. coli, the most efficient compounds were trans-cinnamaldehyde, thymol and thyme. For S. typhi, the most efficient compounds was trans-cinnamaldehyde, carvacrol and thymol. The addition of tetrasodium pyrophosphate, carvacrol and ascorbic acid had no effect on the irradiation sensitivity of E. coli. For S. typhi, only ascorbic acid had no effect.

Differential regulation of caspase-1 activation, pyroptosis, and autophagy via Ipaf and ASC in Shigella-infected macrophages.

Directory of Open Access Journals (Sweden)

Toshihiko Suzuki

2007-08-01

Full Text Available Shigella infection, the cause of bacillary dysentery, induces caspase-1 activation and cell death in macrophages, but the precise mechanisms of this activation remain poorly understood. We demonstrate here that caspase-1 activation and IL-1beta processing induced by Shigella are mediated through Ipaf, a cytosolic pattern-recognition receptor of the nucleotide-binding oligomerization domain (NOD-like receptor (NLR family, and the adaptor protein apoptosis-associated speck-like protein containing a C-terminal caspase recruitment domain (ASC. We also show that Ipaf was critical for pyroptosis, a specialized form of caspase-1-dependent cell death induced in macrophages by bacterial infection, whereas ASC was dispensable. Unlike that observed in Salmonella and Legionella, caspase-1 activation induced by Shigella infection was independent of flagellin. Notably, infection of macrophages with Shigella induced autophagy, which was dramatically increased by the absence of caspase-1 or Ipaf, but not ASC. Autophagy induced by Shigella required an intact bacterial type III secretion system but not VirG protein, a bacterial factor required for autophagy in epithelial-infected cells. Treatment of macrophages with 3-methyladenine, an inhibitor of autophagy, enhanced pyroptosis induced by Shigella infection, suggesting that autophagy protects infected macrophages from pyroptosis. Thus, Ipaf plays a critical role in caspase-1 activation induced by Shigella independently of flagellin. Furthermore, the absence of Ipaf or caspase-1, but not ASC, regulates pyroptosis and the induction of autophagy in Shigella-infected macrophages, providing a novel function for NLR proteins in bacterial-host interactions.

Investigation of Listeria, Salmonella, and toxigenic Escherichia coli in various pet foods.

Science.gov (United States)

Nemser, Sarah M; Doran, Tara; Grabenstein, Michael; McConnell, Terri; McGrath, Timothy; Pamboukian, Ruiqing; Smith, Angele C; Achen, Maya; Danzeisen, Gregory; Kim, Sun; Liu, Yong; Robeson, Sharon; Rosario, Grisel; McWilliams Wilson, Karen; Reimschuessel, Renate

2014-09-01

The Veterinary Laboratory Investigation and Response Network (Vet-LIRN), in collaboration with the Food Emergency Response Network (FERN) and its Microbiology Cooperative Agreement Program (MCAP) laboratories, conducted a study to evaluate the prevalence of selected microbial organisms in various types of pet foods. The goal of this blinded study was to help the Center for Veterinary Medicine prioritize potential future pet food-testing efforts. The study also increased the FERN laboratories' screening capabilities for foodborne pathogens in animal feed matrices, since such pathogens may also be a significant health risk to consumers who come into contact with pet foods. Six U.S. Food and Drug Administration FERN MCAP laboratories analyzed approximately 1056 samples over 2 years. Laboratories tested for Salmonella, Listeria, Escherichia coli O157:H7 enterohemorrhagic E. coli, and Shiga toxin-producing strains of E. coli (STEC). Dry and semimoist dog and cat foods purchased from local stores were tested during Phase 1. Raw dog and cat foods, exotic animal feed, and jerky-type treats purchased through the Internet were tested in Phase 2. Of the 480 dry and semimoist samples, only 2 tested positive: 1 for Salmonella and 1 for Listeria greyii. However, of the 576 samples analyzed during Phase 2, 66 samples were positive for Listeria (32 of those were Listeria monocytogenes) and 15 samples positive for Salmonella. These pathogens were isolated from raw foods and jerky-type treats, not the exotic animal dry feeds. This study showed that raw pet foods may harbor food safety pathogens, such as Listeria monocytogenes and Salmonella. Consumers should handle these products carefully, being mindful of the potential risks to human and animal health.

Quantification of contamination of lettuce by GFP-expressing Escherichia coli O157:H7 and Salmonella enterica serovar Typhimurium

NARCIS (Netherlands)

Franz, Eelco; Visser, Anna A; Van Diepeningen, Anne D; Klerks, Michel M; Termorshuizen, Aad J; van Bruggen, Ariena H C

The primary objective of this study was to determine the possibility of internalization of GFP-expressing Escherichia coli O157:H7 and Salmonella enterica serovar Typhimurium (S. Typhimurium) strains MAE 110 (multi-cellular morphology) and 119 (wild type morphology) into lettuce seedlings (Lactuca

Detection of Extended-Spectrum Β-Lactamases among Gram ...

African Journals Online (AJOL)

Biochemical tests confirmed the identity of the Gram-negative isolates to be members of the enterobactericeae, which included Klebsiella pneumoniae (60), Escherichia coli (98), Providencia Spp. (32), Morganella moganii (32), Shigella Spp. (14), Citrobacter freundii (14), Serratia marcescens (10), Salmonella paratyphi A ...

Intervention technologies for food safety on minimally processed produce:Perspectives on food-borne and plant pathogens

Science.gov (United States)

Produce contamination associated with enteric pathogens such Escherichia coli O157:H7, salmonella spp., Listeria monocytogenes, Shigella and others are significant challenges to food safety. This is due to the illnesses and economic impacts resulting from the outbreaks. Innovative technologies for i...

Potential of predatory bacteria as biocontrol agents for foodborne and plant pathogens

Science.gov (United States)

Foodborne pathogens such as Escherichia coli O157:H7, Salmonella spp., Listeria monocytogenes, Shigella are responsible for frequent occurrences of illnesses and mortality in humans and produce losses. Pre-harvest yield losses and post-harvest decay on minimally processed produce (fruits, vegetables...

Molecular evolution of the nicotinic acid requirement within the Shigella/EIEC pathotype.

Science.gov (United States)

Di Martino, Maria Letizia; Fioravanti, Rosa; Barbabella, Giada; Prosseda, Gianni; Colonna, Bianca; Casalino, Mariassunta

2013-12-01

Nicotinamide adenine dinucleotide (NAD) is a crucial cofactor in several anabolic and catabolic reactions. NAD derives from quinolinic acid (QUIN) which in Escherichia coli is obtained through a pyridine salvage pathway or a de novo synthesis pathway. In the latter case, two enzymes, L-aspartate oxidase (NadB) and quinolinate synthase (NadA), are required for the synthesis of QUIN. In contrast to its E. coli ancestor, Shigella spp., the causative agent of bacillary dissentery, lacks the de novo pathway and strictly requires nicotinic acid for growth (Nic⁻ phenotype). This phenotype depends on the silencing of the nadB and nadA genes and its pathoadaptive nature is suggested by the observation that QUIN attenuates the Shigella invasive process. Shigella shares the pathogenicity mechanism with enteronvasive E. coli (EIEC), a group of pathogenic E. coli. On the basis of this similarity EIEC and Shigella have been grouped into a single E. coli pathotype. However EIEC strains do not constitute a homogeneous group and do not possess the complete set of characters that define Shigella strains. In this work we have analysed thirteen EIEC strains belonging to different serotypes and originating from different geographic areas. We show that, in contrast to Shigella, only some EIEC strains require nicotinic acid for growth in minimal medium. Moreover, by studying the emergence of the Nic⁻ phenotype in all serotypes of S. flexneri, as well as in S. sonnei and S. dysenteriae, we describe which molecular rearrangements occurred and which mutations are responsible for the inactivation of the nadA and nadB genes. Our data confirm that the genome of Shigella is extremely dynamic and support the hypothesis that EIEC might reflect an earlier stage of the pathoadaptation process undergone by Shigella. Copyright © 2013. Published by Elsevier GmbH.

Relationship between Frequency of Escherichia Coli and Prevalence of Salmonella and Shigella Spp. in a Natural River

Directory of Open Access Journals (Sweden)

Ali Shahryari

2017-12-01

Conclusion: Judging the microbial quality of water supplies cannot be sufficient only based on the presence or absence of Escherichia coli bacterium. Therefore, it seems that the use of other secondary indicator bacteria such as fecal streptococci and supplementary sulphite-reducing clostridia will be advised.

Distribution of sulfonamide resistance genes in Escherichia coli and Salmonella isolates from swine and chickens at abattoirs in Ontario and Québec, Canada.

Science.gov (United States)

Kozak, Gosia K; Pearl, David L; Parkman, Julia; Reid-Smith, Richard J; Deckert, Anne; Boerlin, Patrick

2009-09-01

Sulfonamide-resistant Escherichia coli and Salmonella isolates from pigs and chickens in Ontario and Québec were screened for sul1, sul2, and sul3 by PCR. Each sul gene was distributed differently across populations, with a significant difference between distribution in commensal E. coli and Salmonella isolates and sul3 restricted mainly to porcine E. coli isolates.

Occurance and characteristics of class 1, 2 and 3 integrons in Escherichia coli, Salmonella and Campylobacter spp. in the Netherlands

NARCIS (Netherlands)

Essen-Zandbergen, van A.; Smith, H.E.; Veldman, K.T.; Mevius, D.J.

2007-01-01

Objectives: To determine the occurrence and transmission of class 1, 2 and 3 integrons in multidrug-resistant or sulfamethoxazole-resistant Salmonella from human and animal sources and in Campylobacter spp. and Escherichia coli from broilers isolated in the Netherlands in 2004. Methods: PCR,

Extreme Heat Resistance of Food Borne Pathogens Campylobacter jejuni, Escherichia coli, and Salmonella Typhimurium on Chicken Breast Fillet during Cooking

NARCIS (Netherlands)

Jong, de A.E.I.; Asselt, van E.D.; Zwietering, M.H.; Nauta, M.J.; Jonge, de R.

2012-01-01

The aim of this research was to determine the decimal reduction times of bacteria present on chicken fillet in boiling water. The experiments were conducted with Campylobacter jejuni, Salmonella, and Escherichia coli. Whole chicken breast fillets were inoculated with the pathogens, stored overnight

The effect of quorum sensing system for growth competitiveness on Shigella flexneri.

Science.gov (United States)

Xu, Ping; Yang, Jing; Lu, Li-lan; Feng, Er-ling; Wang, Heng-liang; Lu, Ying; Zhu, Li

2015-05-01

Quorum sensing (QS) regulates the onset of bacterial social responses related to cell density. Comparison between the gene sequences of all components of QS system of Escherichia coli and Shigella strains, shows that the QS system is generally lost or mutated in Shigella. Since AI-2 is produced and processed by the lsr operon, we analyzed the potential function of the lsr operon. We first detected AI-2 in Shigella flexneri 2a strain 301 through the reporter bacteria Vibrio harveyi BB170, indicating that S. flexneri can produce AI-2. Then, the lsr operon of E. coli MG1655 was cloned into S. flexneri using the Golden Gate method. Colony counting experiments showed that the QS system recovery strain had growth advantage over the wild-type strain when they were mixed and cultured. The preliminary comparative proteomics analysis showed that the lsr operon could be expressed and the abundance of stress response proteins also changed when the QS system was introduced into S. flexneri.

Shigella outer membrane protein PSSP-1 is broadly protective against Shigella infection.

Science.gov (United States)

Kim, Jae-Ouk; Rho, Semi; Kim, Su Hee; Kim, Heejoo; Song, Hyo Jin; Kim, Eun Jin; Kim, Ryang Yeo; Kim, Eun Hye; Sinha, Anuradha; Dey, Ayan; Yang, Jae Seung; Song, Man Ki; Nandy, Ranjan Kumar; Czerkinsky, Cecil; Kim, Dong Wook

2015-04-01

In developing countries, Shigella is a primary cause of diarrhea in infants and young children. Although antibiotic therapy is an effective treatment for shigellosis, therapeutic options are narrowing due to the emergence of antibiotic resistance. Thus, preventive vaccination could become the most efficacious approach for controlling shigellosis. We have identified several conserved protein antigens that are shared by multiple Shigella serotypes and species. Among these, one antigen induced cross-protection against experimental shigellosis, and we have named it pan-Shigella surface protein 1 (PSSP-1). PSSP-1-induced protection requires a mucosal administration route and coadministration of an adjuvant. When PSSP-1 was administered intranasally, it induced cross-protection against Shigella flexneri serotypes 2a, 5a, and 6, Shigella boydii, Shigella sonnei, and Shigella dysenteriae serotype 1. Intradermally administered PSSP-1 induced strong serum antibody responses but failed to induce protection in the mouse lung pneumonia model. In contrast, intranasal administration elicited efficient local and systemic antibody responses and production of interleukin 17A and gamma interferon. Interestingly, blood samples from patients with recent-onset shigellosis showed variable but significant mucosal antibody responses to other conserved Shigella protein antigens but not to PSSP-1. We suggest that PSSP-1 is a promising antigen for a broadly protective vaccine against Shigella. Copyright © 2015, American Society for Microbiology. All Rights Reserved.

A real-time loop-mediated isothermal amplification assay for rapid detection of Shigella species.

Science.gov (United States)

Liew, P S; Teh, C S J; Lau, Y L; Thong, K L

2014-12-01

Shigellosis is a foodborne illness caused by the genus Shigella and is an important global health issue. The development of effective techniques for rapid detection of this pathogen is essential for breaking the chain of transmission. Therefore, we have developed a novel loop-mediated isothermal amplification (LAMP) assay targeting the invasion plasmid antigen H (ipaH) gene to rapidly detect Shigella species. This assay could be performed in 90 min at an optimal temperature of 64ºC, with endpoint results visualized directly. Notably, the method was found to be more sensitive than conventional PCR. Indeed, the detection limit for the LAMP assay on pure bacterial cultures was 5.9 x 10(5) CFU/ml, while PCR displayed a limit of 5.9 x 10(7) CFU/ml. In spiked lettuce samples, the sensitivity of the LAMP assay was 3.6 x 10(4) CFU/g, whereas PCR was 3.6 x 10(5) CFU/g. Overall, the assay accurately identified 32 Shigella spp. with one enteroinvasive Escherichia coli displaying positive reaction while the remaining 32 non-Shigella strains tested were negative.

Radiosensitivity study of salmonella enteritidis in chickens

International Nuclear Information System (INIS)

Fernandez Gianotti, Tomas

1997-01-01

One of the applications of ionizing radiations in food is the inactivation of vegetative phatogenic bacteria (radicidation) such as Salmonella, Shigella, Campylobacter, Vibro and Listeria. These bacteria are associated with the diseases transmitted by food (ETA). Fresh and frozen farmyard fowls can be contaminated with pathogenic microorganisms, between them Salmonella. In Argentine, between years 1987-1990, Salmonella enteritidis was the main cause of salmonellosis. In food irradiation, with the aim of improving and assuring its hygienic quality, it is important to know the radiosensitivity of microorganisms to be inactivated. Inactivation of a determined microorganism shall depend, between others factors, of the species, strain, number and of the irradiation conditions (temperature, media, etc.). D 10 value is a very useful data in order to compare radiosensitivities between the microorganisms and the influence of different factors in their sensitivities. In this paper, it was determined the sensitivity to the gamma radiation of Salmonella enteritidis in fresh and frozen chickens

Two Cases of Vulvovaginitis Caused by Shigella flexneri and Shigella sonnei: a Case Report

OpenAIRE

Bayramoğlu, Gülçin; Aydın, Faruk; Karagüzel, Gülay; İmamoğlu, Mustafa; Ökten, Ayşenur

2014-01-01

Vulvovaginitis caused by Shigella species (Shigella spp.) has rarely been reported. This paper describes two cases of prepubertal vulvovaginitis, presenting with a bloody and purulent vaginal discharge, separately caused by ampicillin-resistant Shigella flexneri and trimethoprim-sulfomethoxazoleresistant Shigella sonnei. Our conclusions are that Shigella spp. is the potential cause of vulvovaginitis in prepubertal girls in developing countries where these pathogens are endemic, and identifica...

Two Cases of Vulvovaginitis Caused by Shigella flexneri and Shigella sonnei: a Case Report

OpenAIRE

Bayramoğlu, Gülçin; Aydın, Faruk; Karagüzel, Gülay; İmamoğlu, Mustafa; Ökten, Ayşenur

2012-01-01

Vulvovaginitis caused by Shigella species (Shigella spp.) has rarely been reported. This paper describes two cases of prepubertal vulvovaginitis, presenting with a bloody and purulent vaginal discharge, separately caused by ampicillin-resistant Shigella flexneri and trimethoprim-sulfomethoxazoleresistant Shigella sonnei. Our conclusions are that Shigella spp. is the potential cause of vulvovaginitis in prepubertal girls in developing countries where these pathogens are endemic, and identifica...

Agentes bacterianos enteropatogênicos em suínos de diferentes faixas etárias e perfil de resistência a antimicrobianos de cepas de Escherichia coli e Salmonella spp Enteropathogenic bacterial agents in pigs of different age groups and profile of resistance in strains of Escherichia coli and Salmonella spp. to antimicrobial agents

Directory of Open Access Journals (Sweden)

Álvaro Menin

2008-09-01

Full Text Available As enterites infecciosas bacterianas provocam severas perdas para a indústria suína em todo o mundo. Os objetivos deste trabalho foram determinar os agentes bacterianos, associados com a ocorrência de diarréia em suínos, em diferentes faixas etárias, no Estado de Santa Catarina, Brasil, e verificar o perfil de resistência das cepas de Escherichia coli e Salmonella spp, frente aos principais antimicrobianos utilizados em granjas de suínos. Os principais gêneros/espécies bacterianos diagnosticados foram Escherichia coli, Clostridium spp, Salmonella spp Brachyspira hyodysenteriae, Brachyspira pilosicoli e Lawsonia intracellularis. Os fatores de virulência de E. coli mais prevalentes na fase de maternidade foram F5 / (K99 20%, F6 / (987P 16,3%, F42 6,8% e F41 5,7%, já nas fases de creche e terminação, predominaram cepas com fimbrias F4 (K88 11,2% e 5,4%, respectivamente. Para E. coli os maiores índices de resistência foram encontrados para oxitetraciclina (94% e tetraciclina (89,5% e os menores índices de resistência para neomicina (55%, ceftiofur (57,4%. Quanto às amostras de Salmonella spp, estas apresentaram maior resistência à oxitetraciclina (77%, e à tetraciclina (42,1% e menor à gentamicina (3,5% e amoxicilina (4,8%.Infectious bacterial enteritis causes severe losses to the swine industry worldwide. The objective of this study was to determine the epidemiology of bacterial agents that are associated with the occurrence of diarrhea in pigs at different age groups, and to verify the profile of resistance of strains of Escherichia coli and Salmonella spp to the main antimicrobial agents. The main bacterial species diagnosed were Escherichia coli, Clostridium spp, Salmonella spp, Brachyspira hyodysenteriae, Brachyspira pilosicoli and Lawsonia intracellularis. The E. coli virulence factors of higher prevalence in preweaning piglets were F5 / (K99 20%, F6 / (987P 16.3%, F42 6.8% and F41 5.7%, whereas at the nursery and with

Phagocytic and chemiluminescent responses of mouse peritoneal macrophages to living and killed Salmonella typhimurium and other bacteria

International Nuclear Information System (INIS)

Tomita, T.; Blumenstock, E.; Kanegasaki, S.

1981-01-01

In the presence of luminol, resident as well as thioglycolate-induced and immunized macrophages emitted chemiluminescence more efficiently when the cells were exposed to living Salmonella typhimurium than when they were exposed to the same bacterium killed by ultraviolet light or heat. This phenomenon was observed whether or not the bacterium was opsonized. The different response to living and killed bacteria was also found with Escherichia coli, Pseudomonas aeruginosa, Proteus morganii, and Enterobacter aerogenes, but not with Shigella sonnei, Klebsiella pneumoniae, and Propionibacterium acnes. The results suggest that macrophages respond better to living, motile bacteria than to nonmotile or killed bacteria. The experimental results obtained with motility mutants of S. typhimurium, E. coli, and P. aeruginosa confirm that macrophages exposed to the motile bacteria emit chemiluminescence more efficiently and ingest the motile bacteria at a much faster rate than the nonmotile bacteria

Antimicrobial activity of traditional wines (Sopi and Moke against Salmonella sp. and Escherichia coli

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Annytha Ina Rohi Detha

2016-09-01

Full Text Available Objective: Sopi and Moke are two traditional wines in Indonesia. The present study aimed at investigating the antimicrobial effects of Sopi and Moke as compared to other commercial disinfectants. Materials and methods: The alcohol level and pH of the traditional wines (Sopi and Moke were determined by alcohol meter and pH meter, respectively. The susceptibility test was perfomed to determine the antimicrobial activity of Sopi against Escherichia coli which was isolated from cattle, and the activity of Moke was tested against Salmonella sp. which was a local isolate of poultry. Results: In susceptibility test, Sopi showed 17.5 mm in zone of inhibition against E. coli, while Formades and reg;, a commercial disinfectant showed 16 mm of zone of inhibition against the same bacteria. Moke showed 17.5 mm inhibition zone against Salmonella sp., whereas Antisep and reg;, a commercial disinfectant had 28 mm of inhibition zone against the same isolate. Conclusion: The results indicate that Sopi and Moke have antimicrobial effects on E. coli and Salmonella sp., respectively. The findings of this study suggest that Sopi and Moke can be used as potential antimicrobial agents. [J Adv Vet Anim Res 2016; 3(3.000: 282-285

Composition for the treatment of radiation sickness and topical bacterial infections

International Nuclear Information System (INIS)

Gaffin, S.L.

1985-01-01

A therapeutic composition for the theatment of radiation sickness comprises a carrier medium including one or more antibodies specific to gram negative bacteria chosen from the group Klebsiella, Pseudomoneas, Shigella, Salmonella and Escherichia species is discussed. The composition may by formed for oral administration or topical application

Epidemiology of bacterial pathogens associated with infectious diarrhea in Djibouti.

Science.gov (United States)

Mikhail, I A; Fox, E; Haberberger, R L; Ahmed, M H; Abbatte, E A

1990-01-01

During a survey examining the causes of diarrhea in the East African country of Djibouti, 140 bacterial pathogens were recovered from 209 diarrheal and 100 control stools. The following pathogens were isolated at comparable frequencies from both diarrheal and control stools: enteroadherent Escherichia coli (EAEC) (10.6 versus 13%), enterotoxigenic E. coli (ETEC) (11 versus 10%), enteropathogenic E. coli (EPEC) (7.7 versus 12%), Salmonella spp. (2.9 versus 3%), and Campylobacter jejuni-C. coli (3.3 versus 5%). Surprisingly, the EAEC strains isolated did not correspond to well-recognized EPEC serogroups. No Yersinia spp., enteroinvasive E. coli, or enterohemorrhagic E. coli were isolated during the course of this study. Only the following two genera were recovered from diarrheal stools exclusively: Shigella spp. (7.7%) and Aeromonas hydrophila group organisms (3.3%). Shigella flexneri was the most common Shigella species isolated. Patients with Shigella species were of a higher average age than were controls (27 versus 13 years), while subjects with Campylobacter or Salmonella species belonged to younger age groups (2.6 and 1.6 years, respectively). Salmonella cases were more often in females. Shigella diarrhea was associated with fecal blood or mucus and leukocytes. ETEC was not associated with nausea or vomiting. Anorexia, weight loss, and fever were associated with the isolation of Salmonella and Aeromonas species. EAEC, ETEC, EPEC, and Shigella species were resistant to most drugs used for treating diarrhea in Africa, while the antibiotic most active against all bacteria tested was norfloxacin. We conclude that in Djibouti in 1989, Shigella and Aeromonas species must be considered as potential pathogens whenever they are isolated from diarrheal stools and that norfloxacin should be considered the drug of choice in adults for treating severe shigellosis and for diarrhea prophylaxis in travelers. PMID:2351738

Distribution of Sulfonamide Resistance Genes in Escherichia coli and Salmonella Isolates from Swine and Chickens at Abattoirs in Ontario and Québec, Canada ▿

Science.gov (United States)

Kozak, Gosia K.; Pearl, David L.; Parkman, Julia; Reid-Smith, Richard J.; Deckert, Anne; Boerlin, Patrick

2009-01-01

Sulfonamide-resistant Escherichia coli and Salmonella isolates from pigs and chickens in Ontario and Québec were screened for sul1, sul2, and sul3 by PCR. Each sul gene was distributed differently across populations, with a significant difference between distribution in commensal E. coli and Salmonella isolates and sul3 restricted mainly to porcine E. coli isolates. PMID:19633109

Development of an Aotus nancymaae Model for Shigella Vaccine Immunogenicity and Efficacy Studies

Science.gov (United States)

Gregory, Michael; Lugo-Roman, Luis A.; Galvez Carrillo, Hugo; Tilley, Drake Hamilton; Baldeviano, Christian; Simons, Mark P.; Reynolds, Nathanael D.; Ranallo, Ryan T.; Suvarnapunya, Akamol E.; Venkatesan, Malabi M.; Oaks, Edwin V.

2014-01-01

Several animal models exist to evaluate the immunogenicity and protective efficacy of candidate Shigella vaccines. The two most widely used nonprimate models for vaccine development include a murine pulmonary challenge model and a guinea pig keratoconjunctivitis model. Nonhuman primate models exhibit clinical features and gross and microscopic colonic lesions that mimic those induced in human shigellosis. Challenge models for enterotoxigenic Escherichia coli (ETEC) and Campylobacter spp. have been successfully developed with Aotus nancymaae, and the addition of a Shigella-Aotus challenge model would facilitate the testing of combination vaccines. A series of experiments were designed to identify the dose of Shigella flexneri 2a strain 2457T that induces an attack rate of 75% in the Aotus monkey. After primary challenge, the dose required to induce an attack rate of 75% was calculated to be 1 × 1011 CFU. Shigella-specific immune responses were low after primary challenge and subsequently boosted upon rechallenge. However, preexisting immunity derived from the primary challenge was insufficient to protect against the homologous Shigella serotype. A successive study in A. nancymaae evaluated the ability of multiple oral immunizations with live-attenuated Shigella vaccine strain SC602 to protect against challenge. After three oral immunizations, animals were challenged with S. flexneri 2a 2457T. A 70% attack rate was demonstrated in control animals, whereas animals immunized with vaccine strain SC602 were protected from challenge (efficacy of 80%; P = 0.05). The overall study results indicate that the Shigella-Aotus nancymaae challenge model may be a valuable tool for evaluating vaccine efficacy and investigating immune correlates of protection. PMID:24595138

Evaluation of Eight Different Cephalosporins for Detection of Cephalosporin Resistance in Salmonella enterica and Escherichia coli

DEFF Research Database (Denmark)

Aarestrup, Frank Møller; Hasman, Henrik; Veldman, K

2010-01-01

This study evaluates the efficacy of eight different cephalosporins for detection of cephalosporin resistance mediated by extended spectrum beta-lactamases (ESBL) and plasmidic AmpC beta-lactamases in Salmonella and Escherichia coli. A total of 138 E. coli and 86 Salmonella isolates with known beta......-resistant but cephalosporin-susceptible, 56 ESBL isolates and 19 isolates with plasmidic AmpC, as well as 10 ampC hyper-producing E. coli. The minimum inhibitory concentration distributions and zone inhibitions varied with the tested compound. Ampicillin-resistant isolates showed reduced susceptibility to the cephalosporins...... compared to ampicillin-susceptible isolates. Cefoperazone, cefquinome, and cefuroxime were not useful in detecting isolates with ESBL or plasmidic AmpC. The best substances for detection were cefotaxime, cefpodoxime, and ceftriaxone, whereas ceftazidime and ceftiofur were not as efficient. Ceftriaxone may...

Effects of thermosonication on the fate of Escherichia coli O157:H7 and Salmonella Enteritidis in mango juice.

Science.gov (United States)

Kiang, W-S; Bhat, R; Rosma, A; Cheng, L-H

2013-04-01

In this study, the effects of thermosonication and thermal treatment on Escherichia coli O157:H7 and Salmonella Enteritidis in mango juice were investigated at 50 and 60°C. Besides, nonlethal injury of Salm. Enteritidis after both treatments was also examined. The highest inactivation was attained with thermosonication at 60°C. The inactivation rate was different for both pathogens, and Salm. Enteritidis was found to be more sensitive to thermosonication than E. coli O157:H7. Salmonella Enteritidis was recovered in all treated samples, except those subjected to more than 5-min thermosonication at 60°C. It was found that the introduction of high-intensity ultrasound enhanced the inactivation of pathogens compared to thermal treatment alone. On the other hand, Salm. Enteritidis was detected in a number of samples following incubation in universal pre-enrichment broth, but no growth was detected after incubation in mango juice. Fruit juices are commonly heat treated to inactivate micro-organisms and enzymes. However, excessive heat treatments may result in undesirable changes in juice quality. Treatment by power ultrasound, a nonthermal technology, may be an alternative processing technique to pasteurize fruit juices. This study highlights the effectiveness of thermosonication in inactivating Escherichia coli O157:H7 and Salmonella Enteritidis in mango juice. © 2012 The Society for Applied Microbiology.

Effect of Salvia chorassanica Root Aqueous, Ethanolic and Hydro Alcoholic Extracts on Staphylococcus aureus, Enterococcus faecalis, Salmonella typhimurium and Escherichia coli

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Azam Mehraban

2016-11-01

Full Text Available Background Nowadays, through the previous researches, it has become clear that Salvia has important health benefits. Salvia chorassanica is one of the valuable native Iranian species which only grows in Khorasan province, Iran. Objectives The aim of this study is to evaluate the antimicrobial effect of Salvia chorassanica root aqueous, ethanolic and hydro alcoholic extracts on Staphylococcus aureus, Enterococcus faecalis, Salmonella typhimurium and Escherichia coli. Methods In this experimental study, maceration method was used to prepare extracts. Study setup was conducted in March 2014.The duration of study setup took for two months. The micro dilution method by ELISA was used to determine the minimum inhibitory concentration (MIC and minimum bactericidal concentration (MBC of aqueous, ethanolic and hydro alcoholic extracts of root of Salvia chorassanica against Staphylococcus aureus, Enterococcus faecalis, Salmonella typhimurium and Escherichia coli. The antibacterial effect also was evaluated using agar diffusion method. The inhibition zones of growth against the extracts were measured in comparison to standards antibiotics. Chloramphenicol as positive control on Enterococcus faecalis, Tetracycline on Staphylococcus aureus, Gentamicin on Escherichia coli and Neomycin on Salmonella typhimurium. The data were analyzed using one way analysis of variance (ANOVA with SPSS version 16. Results The highest inhibition zone in diffusion method was related to ethanolic extract of Salvia chorassanica root against Gram-positive bacteria, Staphylococcus aureus and Enterococcus faecalis. The calculated MIC in aqueous and ethanolic extracts of root for Staphylococcus aureus was 240 and 120 mg/mL, for Enterococcus faecalis was 120 and 60 mg/mL respectively, and for Escherichia coli and Salmonella typhimurium was equal to 240 mg/mL. The amount in hydro alcoholic extracts for Gram-positive bacteria was 60 mg/mL and for Gram-negative bacteria was 120 mg/mL. The

Dielectric barrier discharge atmospheric cold plasma inhibits Escherichia coli 0157:H7, Salmonella, Listeria monocytogenes, and Tulane virus in Romaine lettuce

Science.gov (United States)

The present study investigated the effects of dielectric barrier discharge atmospheric cold plasma (DACP) treatment on the inactivation of Escherichia coli O157:H7, Salmonella, Listeria monocytogenes, and Tulane virus (TV) on Romaine lettuce, assessing the influences of moisture vaporization, modifi...

Sensitivity and specificity of leukocyte count in feces as a predictor of stool culture positivity for S amonella or Shigella Sensibilidad y especificidad del recuento de leucocitos en las materias fecales para predecir la presencia de Salmonella o Shigella en pacientes con enfermedad diarreica aguda

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John Jairo Zuleta Tobón

2008-01-01

Full Text Available

In most studies of fecal leukocyte counts as predictors of the result of stool cultures the sensitivity and specificity were not determined.

Objective: to evaluate fecal leukocyte counts as a predictor of the presence of Salmonella and Shigella.

Design: a descriptive, cross section study was carried out in 905 stool cultures at a university hospital in Medellín, Colombia. Results for Salmonella and Shigella were taken as the gold standard to evaluate the sensitivity and specificity of fecal leukocyte counts.

Results: sensitivity and specificity, according to the level of the count, were, respectively: 1-5 leukocytes: 89.2% and 57.1%; 6-10 leukocytes: 86.2% and 52.8%;11-20 leukocytes: 77.7% and 62.7%; 21-30 leukocytes: 63.9% and 76.3%; 31-50 leukocytes: 45.2% and 85.5%; more than 50 leukocytes: 28.3% and 90.9%. The area under the curve was 0.7699 (CI 95%: 0.7275-0.8123.

Conclusions: Polymorphism of CRISPR shows separated natural groupings of Shigella subtypes and evidence of horizontal transfer of CRISPR

Science.gov (United States)

Yang, Chaojie; Li, Peng; Su, Wenli; Li, Hao; Liu, Hongbo; Yang, Guang; Xie, Jing; Yi, Shengjie; Wang, Jian; Cui, Xianyan; Wu, Zhihao; Wang, Ligui; Hao, Rongzhang; Jia, Leili; Qiu, Shaofu; Song, Hongbin

2015-01-01

Clustered, regularly interspaced, short palindromic repeats (CRISPR) act as an adaptive RNA-mediated immune mechanism in bacteria. They can also be used for identification and evolutionary studies based on polymorphisms within the CRISPR locus. We amplified and analyzed 6 CRISPR loci from 237 Shigella strains belonging to the 4 species groups, as well as 13 Escherichia coli strains. The CRISPR-associated (cas) gene sequence arrays of these strains were screened and compared. The CRISPR sequences from Shigella were conserved among subtypes, suggesting that CRISPR may represent a new identification tool for the detection and discrimination of Shigella species. Secondary structure analysis showed a different stem-loop structure at the terminal repeat, suggesting a distinct recognition mechanism in the formation of crRNA. In addition, the presence of “self-target” spacers and polymorphisms within CRISPR in Shigella indicated a selective pressure for inhibition of this system, which has the potential to damage “self DNA.” Homology analysis of spacers showed that CRISPR might be involved in the regulation of virulence transmission. Phylogenetic analysis based on CRISPR sequences from Shigella and E. coli indicated that although phenotypic properties maintain convergent evolution, the 4 Shigella species do not represent natural groupings. Surprisingly, comparative analysis of Shigella repeats with other species provided new evidence for CRISPR horizontal transfer. Our results suggested that CRISPR analysis is applicable for the detection of Shigella species and for investigation of evolutionary relationships. PMID:26327282

Polymorphism of CRISPR shows separated natural groupings of Shigella subtypes and evidence of horizontal transfer of CRISPR.

Science.gov (United States)

Yang, Chaojie; Li, Peng; Su, Wenli; Li, Hao; Liu, Hongbo; Yang, Guang; Xie, Jing; Yi, Shengjie; Wang, Jian; Cui, Xianyan; Wu, Zhihao; Wang, Ligui; Hao, Rongzhang; Jia, Leili; Qiu, Shaofu; Song, Hongbin

2015-01-01

Clustered, regularly interspaced, short palindromic repeats (CRISPR) act as an adaptive RNA-mediated immune mechanism in bacteria. They can also be used for identification and evolutionary studies based on polymorphisms within the CRISPR locus. We amplified and analyzed 6 CRISPR loci from 237 Shigella strains belonging to the 4 species groups, as well as 13 Escherichia coli strains. The CRISPR-associated (cas) gene sequence arrays of these strains were screened and compared. The CRISPR sequences from Shigella were conserved among subtypes, suggesting that CRISPR may represent a new identification tool for the detection and discrimination of Shigella species. Secondary structure analysis showed a different stem-loop structure at the terminal repeat, suggesting a distinct recognition mechanism in the formation of crRNA. In addition, the presence of "self-target" spacers and polymorphisms within CRISPR in Shigella indicated a selective pressure for inhibition of this system, which has the potential to damage "self DNA." Homology analysis of spacers showed that CRISPR might be involved in the regulation of virulence transmission. Phylogenetic analysis based on CRISPR sequences from Shigella and E. coli indicated that although phenotypic properties maintain convergent evolution, the 4 Shigella species do not represent natural groupings. Surprisingly, comparative analysis of Shigella repeats with other species provided new evidence for CRISPR horizontal transfer. Our results suggested that CRISPR analysis is applicable for the detection of Shigella species and for investigation of evolutionary relationships.

Multiple antimicrobial resistance of Escherichia coli and Salmonella ...

African Journals Online (AJOL)

Presumptive isolates were subjected to antimicrobial susceptibility testing using 13 panels of antibiotics for both E. coli and Salmonella spp. Results showed that the overall isolation rate of Salmonella spp. was 12 (11.4%), broiler chickens had higher isolation rate 9 (12.0%) of Salmonella than local chickens. However, the ...

Two Cases of Vulvovaginitis Caused by Shigella flexneri and Shigella sonnei: a Case Report

Directory of Open Access Journals (Sweden)

Gülçin Bayramoğlu

2012-03-01

Full Text Available Vulvovaginitis caused by Shigella species (Shigella spp. has rarely been reported. This paper describes two cases of prepubertal vulvovaginitis, presenting with a bloody and purulent vaginal discharge, separately caused by ampicillin-resistant Shigella flexneri and trimethoprim-sulfomethoxazole-resistant Shigella sonnei. Our conclusions are that Shigella spp. is the potential cause of vulvovaginitis in prepubertal girls in developing countries where these pathogens are endemic, and identification of the bacteria and making antibiotic susceptibility testing in these cases should not be overlooked.

Pathogenicity of Shigella in chickens.

Science.gov (United States)

Shi, Run; Yang, Xia; Chen, Lu; Chang, Hong-tao; Liu, Hong-ying; Zhao, Jun; Wang, Xin-wei; Wang, Chuan-qing

2014-01-01

Shigellosis in chickens was first reported in 2004. This study aimed to determine the pathogenicity of Shigella in chickens and the possibility of cross-infection between humans and chickens. The pathogenicity of Shigella in chickens was examined via infection of three-day-old SPF chickens with Shigella strain ZD02 isolated from a human patient. The virulence and invasiveness were examined by infection of the chicken intestines and primary chicken intestinal epithelial cells. The results showed Shigella can cause death via intraperitoneal injection in SPF chickens, but only induce depression via crop injection. Immunohistochemistry and transmission electron microscopy revealed the Shigella can invade the intestinal epithelia. Immunohistochemistry of the primary chicken intestinal epithelial cells infected with Shigella showed the bacteria were internalized into the epithelial cells. Electron microscopy also confirmed that Shigella invaded primary chicken intestinal epithelia and was encapsulated by phagosome-like membranes. Our data demonstrate that Shigella can invade primary chicken intestinal epithelial cells in vitro and chicken intestinal mucosa in vivo, resulting in pathogenicity and even death. The findings suggest Shigella isolated from human or chicken share similar pathogenicity as well as the possibility of human-poultry cross-infection, which is of public health significance.

Avaliação da presença de microrganismos indicadores higiênico-sanitários em alimentos servidos em escolas públicas de Porto Alegre, Brasil Evaluation of the presence of hygienic and sanitary indicator microorganisms in food served in public schools in Porto Alegre, Brazil

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Marisa Ribeiro de Itapema Cardoso

2013-04-01

Full Text Available Este estudo tem por objetivo avaliar a presença de microrganismos indicadores higiênico-sanitários em amostras de alimentos servidos em escolas públicas de Porto Alegre. Foram analisados todos os alimentos servidos na refeição do turno da visita, quanto à presença de Escherichia coli, Staphylococcus coagulase positiva, Salmonella sp. e Shigella sp. No total de 196 alimentos analisados de 120 escolas, 4 apresentavam contagem de Escherichia coli acima do permitido e dois tinham a presença de Staphylococcus coagulase positiva. Os gêneros Shigella e Salmonella não foram encontrados. Foi observado que a maioria das escolas estudadas servia alimentos dentro de padrões higiênico-sanitários adequados. Foi evidenciado que somente escolas municipais contavam com a orientação de responsável técnico pela alimentação escolar. Das escolas estaduais 60% nunca haviam recebido visita de nutricionista nas quais foram encontrados procedimentos em desacordo com as exigências da legislação. Na maioria das escolas, os alimentos servidos estavam dentro de padrões adequados, porém os problemas detectados demonstram a necessidade da implantação das Boas Práticas no ambiente escolar.The objective of this study was to evaluate the presence of hygienic and sanitary indicator microorganisms in samples of food served in public schools in Porto Alegre. All the food served in the meal of the session visited was analyzed for Escherichia coli, coagulase-positive Staphylococcus, Salmonella sp. and Shigella sp. Of the total of 196 food products analyzed in 120 schools, 4 contained and Escherichia coli score above the permitted level, and 2 contained coagulase-positive Staphylococcus. Neither Shigella nor Salmonella genus were detected. In the majority of schools studied, it was found that food was of an adequate hygienic-sanitary standard. However, only municipal schools had the supervision of a technician responsible for school food. In the state

Multiplex polymerase chain reaction for identification of Escherichia coli, Escherichia albertii and Escherichia fergusonii.

Science.gov (United States)

Lindsey, Rebecca L; Garcia-Toledo, L; Fasulo, D; Gladney, L M; Strockbine, N

2017-09-01

Escherichia coli, Escherichia albertii, and Escherichia fergusonii are closely related bacteria that can cause illness in humans, such as bacteremia, urinary tract infections and diarrhea. Current identification strategies for these three species vary in complexity and typically rely on the use of multiple phenotypic and genetic tests. To facilitate their rapid identification, we developed a multiplex PCR assay targeting conserved, species-specific genes. We used the Daydreamer™ (Pattern Genomics, USA) software platform to concurrently analyze whole genome sequence assemblies (WGS) from 150 Enterobacteriaceae genomes (107 E. coli, 5 Shigella spp., 21 E. albertii, 12 E. fergusonii and 5 other species) and design primers for the following species-specific regions: a 212bp region of the cyclic di-GMP regulator gene (cdgR, AW869_22935 from genome K-12 MG1655, CP014225) for E. coli/Shigella; a 393bp region of the DNA-binding transcriptional activator of cysteine biosynthesis gene (EAKF1_ch4033 from genome KF1, CP007025) for E. albertii; and a 575bp region of the palmitoleoyl-acyl carrier protein (ACP)-dependent acyltransferase (EFER_0790 from genome ATCC 35469, CU928158) for E. fergusonii. We incorporated the species-specific primers into a conventional multiplex PCR assay and assessed its performance with a collection of 97 Enterobacteriaceae strains. The assay was 100% sensitive and specific for detecting the expected species and offers a quick and accurate strategy for identifying E. coli, E. albertii, and E. fergusonii in either a single reaction or by in silico PCR with sequence assemblies. Published by Elsevier B.V.

Detecção de fatores de virulência de Escherichia coli e análise de Salmonella spp. em psitacídeos

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Isadora M. de O. Corrêa

2013-02-01

Full Text Available A flora entérica dos psitacídeos é composta principalmente por bactérias Gram positivas. Bactérias Gram negativas, como Escherichia coli e Salmonella spp., apresentam elevado potencial patogênico, sendo consideradas indicativo de problemas de manejo, que poderão culminar em manifestação de doenças em decorrência de fatores estressantes, dietas deficientes e superlotação, combinados com alta carga bacteriana no ambiente. O objetivo deste trabalho foi avaliar a presença de Salmonella spp., Escherichia coli e os fatores de virulência dos genes iss e iutA dos isolados de E. coli. Analisou-se um total de 44 amostras provenientes de psitacídeos criados em cativeiro, sendo estas 15 fragmentos de órgãos de aves submetidas a exame de necropsia e também 29 amostras de swabs de cloaca e inglúvio de papagaios-charão (Amazona pretrei criados em cativeiro. Nenhuma amostra foi positiva para Salmonella spp. Nas amostras de E. coli detectou-se ambos os fatores de virulência pesquisados.

Transferability of antimicrobial resistance from multidrug-resistant Escherichia coli isolated from cattle in the USA to E. coli and Salmonella Newport recipients

Science.gov (United States)

The objective of this study was to evaluate conjugative transfer of cephalosporin resistance among (n=100) strains of multi-drug resistant Escherichia coli (MDRE) to Salmonella Newport and E. coli DH5-alpha recipients. To accomplish this, phenotypic and genotypic profiles were determined for MDRE, ...

Risk Factors for Salmonella, Shiga Toxin-Producing Escherichia coli and Campylobacter Occurrence in Primary Production of Leafy Greens and Strawberries

Directory of Open Access Journals (Sweden)

Siele Ceuppens

2015-08-01

Full Text Available The microbiological sanitary quality and safety of leafy greens and strawberries were assessed in the primary production in Belgium, Brazil, Egypt, Norway and Spain by enumeration of Escherichia coli and detection of Salmonella, Shiga toxin-producing E. coli (STEC and Campylobacter. Water samples were more prone to containing pathogens (54 positives out of 950 analyses than soil (16/1186 and produce on the field (18/977 for leafy greens and 5/402 for strawberries. The prevalence of pathogens also varied markedly according to the sampling region. Flooding of fields increased the risk considerably, with odds ratio (OR 10.9 for Salmonella and 7.0 for STEC. A significant association between elevated numbers of generic E. coli and detection of pathogens (OR of 2.3 for STEC and 2.7 for Salmonella was established. Generic E. coli was found to be a suitable index organism for Salmonella and STEC, but to a lesser extent for Campylobacter. Guidelines on frequency of sampling and threshold values for E. coli in irrigation water may differ from region to region.

Serogroup prevalence of Shigellae in Bombay.

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Sonawala M

1995-10-01

Full Text Available Prevalence of Shigellae serotypes in Bombay was studied from June 1988 to May 1991. A total of 2758 faecal specimens were collected from paediatric patients (< 12 yrs with acute gastroenteritis. A total of 90 Shigella were isolated giving the isolation rate of 3.2%. Shigella flexneri was the predominant serogroup (73.3% followed by Shigella dysenteriae (16.6%. All the isolates were sensitive to nalidixic acid. Eighty percent of the Shigellae were multidrug resistant. Present data were compared with the study carried out during the period of 1983-87 from the same institute. A change in the serogroup prevalence was noted wherein Shigella flexneri dominated over Shigella dysenteriae since 1985. Increase in resistance to ampicillin and cotrimoxazole was seen in Shigella flexneri strains as compared to previous years.

Penerapan Metode Dempster Shafer Untuk Mendiagnosa Penyakit Dari Akibat Bakteri Salmonella

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Mikha Dayan Sinaga

2016-12-01

Full Text Available Infeksi dari bakteri Samonella dapat menyerang saluran gastrointestin yang mencakup perut, usus halus, dan usus besar atau kolon. Beberapa spesies salmonella dapat menyebabkan infeksi melalui makanan. Termasuk ke dalamnya adalah Salmonella Typhi yang mengakibatkan penyakit tifus, dan Salmonella Shigella yang mengakibatkan penyakit disentri dan diare. Masih banyak orang yang belum mengetahui gejala-gejala dari infeksi bakteri ini serta bagaimana cara untuk mendiagnosa dengan nilai kepastian yang tinggi. Untuk dapat mengetahui tingkat kepastian infeksi bakteri ini peneliti menggunakan metode Dempster-Shafer. Metode ini dipilih karena metode ini dianggap mampu untuk memberikan tingkat kepastian yang tinggi. Metode Dempster-Shafer adalah representasi, kombinasi dan propogasi ketidakpastian, dimana teori ini memiliki beberapa karakteristik yang secara instutif sesuai dengan cara berfikir seorang pakar, namun dasar matematika yang kuat. Hasil dari penelitian ini adalah untuk membuat aplikasi sistem pakar yang dapat mendiagnosa bakteri dari akibat bakteri salmonella dengan menggunakan metode Dempster Shafer. Kata Kunci  : Sistem Pakar,Metode Dempster Shafer, Bakteri Salmonella.

The small, slow and specialized CRISPR and anti-CRISPR of Escherichia and Salmonella.

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Marie Touchon

Full Text Available Prokaryotes thrive in spite of the vast number and diversity of their viruses. This partly results from the evolution of mechanisms to inactivate or silence the action of exogenous DNA. Among these, Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR are unique in providing adaptive immunity against elements with high local resemblance to genomes of previously infecting agents. Here, we analyze the CRISPR loci of 51 complete genomes of Escherichia and Salmonella. CRISPR are in two pairs of loci in Escherichia, one single pair in Salmonella, each pair showing a similar turnover rate, repeat sequence and putative linkage to a common set of cas genes. Yet, phylogeny shows that CRISPR and associated cas genes have different evolutionary histories, the latter being frequently exchanged or lost. In our set, one CRISPR pair seems specialized in plasmids often matching genes coding for the replication, conjugation and antirestriction machinery. Strikingly, this pair also matches the cognate cas genes in which case these genes are absent. The unexpectedly high conservation of this anti-CRISPR suggests selection to counteract the invasion of mobile elements containing functional CRISPR/cas systems. There are few spacers in most CRISPR, which rarely match genomes of known phages. Furthermore, we found that strains divergent less than 250 thousand years ago show virtually identical CRISPR. The lack of congruence between cas, CRISPR and the species phylogeny and the slow pace of CRISPR change make CRISPR poor epidemiological markers in enterobacteria. All these observations are at odds with the expectedly abundant and dynamic repertoire of spacers in an immune system aiming at protecting bacteria from phages. Since we observe purifying selection for the maintenance of CRISPR these results suggest that alternative evolutionary roles for CRISPR remain to be uncovered.

Antimicrobial activities of red wine-based formulations containing plant extracts against Escherichia coli O157:H7 and Salmonella enterica serovar Hadar

Science.gov (United States)

We evaluated the bactericidal activities of red wine and red wine solutions containing a commercial olive extract called Hydrox-12, oregano oil, or a mixture of both in red wine against the following foodborne pathogens: Escherichia coli O157:H7, Salmonella enterica, Listeria monocytogenes, and Stap...

An attenuated Shigella mutant lacking the RNA-binding protein Hfq provides cross-protection against Shigella strains of broad serotype.

Science.gov (United States)

Mitobe, Jiro; Sinha, Ritam; Mitra, Soma; Nag, Dhrubajyoti; Saito, Noriko; Shimuta, Ken; Koizumi, Nobuo; Koley, Hemanta

2017-07-01

Few live attenuated vaccines protect against multiple serotypes of bacterial pathogen because host serotype-specific immune responses are limited to the serotype present in the vaccine strain. Here, immunization with a mutant of Shigella flexneri 2a protected guinea pigs against subsequent infection by S. dysenteriae type 1 and S. sonnei strains. This deletion mutant lacked the RNA-binding protein Hfq leading to increased expression of the type III secretion system via loss of regulation, resulting in attenuation of cell viability through repression of stress response sigma factors. Such increased antigen production and simultaneous attenuation were expected to elicit protective immunity against Shigella strains of heterologous serotypes. Thus, the vaccine potential of this mutant was tested in two guinea pig models of shigellosis. Animals vaccinated in the left eye showed fewer symptoms upon subsequent challenge via the right eye, and even survived subsequent intestinal challenge. In addition, oral vaccination effectively induced production of immunoglobulins without severe side effects, again protecting all animals against subsequent intestinal challenge with S. dysenteriae type 1 or S. sonnei strains. Antibodies against common virulence proteins and the O-antigen of S. flexneri 2a were detected by immunofluorescence microscopy. Reaction of antibodies with various strains, including enteroinvasive Escherichia coli, suggested that common virulence proteins induced protective immunity against a range of serotypes. Therefore, vaccination is expected to cover not only the most prevalent serotypes of S. sonnei and S. flexneri 2a, but also various Shigella strains, including S. dysenteriae type 1, which produces Shiga toxin.

An attenuated Shigella mutant lacking the RNA-binding protein Hfq provides cross-protection against Shigella strains of broad serotype.

Directory of Open Access Journals (Sweden)

Jiro Mitobe

2017-07-01

Full Text Available Few live attenuated vaccines protect against multiple serotypes of bacterial pathogen because host serotype-specific immune responses are limited to the serotype present in the vaccine strain. Here, immunization with a mutant of Shigella flexneri 2a protected guinea pigs against subsequent infection by S. dysenteriae type 1 and S. sonnei strains. This deletion mutant lacked the RNA-binding protein Hfq leading to increased expression of the type III secretion system via loss of regulation, resulting in attenuation of cell viability through repression of stress response sigma factors. Such increased antigen production and simultaneous attenuation were expected to elicit protective immunity against Shigella strains of heterologous serotypes. Thus, the vaccine potential of this mutant was tested in two guinea pig models of shigellosis. Animals vaccinated in the left eye showed fewer symptoms upon subsequent challenge via the right eye, and even survived subsequent intestinal challenge. In addition, oral vaccination effectively induced production of immunoglobulins without severe side effects, again protecting all animals against subsequent intestinal challenge with S. dysenteriae type 1 or S. sonnei strains. Antibodies against common virulence proteins and the O-antigen of S. flexneri 2a were detected by immunofluorescence microscopy. Reaction of antibodies with various strains, including enteroinvasive Escherichia coli, suggested that common virulence proteins induced protective immunity against a range of serotypes. Therefore, vaccination is expected to cover not only the most prevalent serotypes of S. sonnei and S. flexneri 2a, but also various Shigella strains, including S. dysenteriae type 1, which produces Shiga toxin.

Effectiveness of different antimicrobial washes combined with freezing against Escherichia coli O157:H7, Salmonella Typhimurium, and Listeria monocytogenes inoculated on blueberries

Science.gov (United States)

To ensure the microbial safety of produce including blueberries, sanitization is a critical step. This study evaluated the efficacy of sanitizers when coupled with frozen storage, in inactivating Escherichia coli O157:H7, Salmonella Typhimurium and Listeria monocytogenes inoculated on wild blueberri...

ANTIMICIROBIAL SUSCEPTIBILITY PATTERNS OF Escherichia coli ...

African Journals Online (AJOL)

DR. AMINU

A total of 56 and 24 strains of E. coli and Shigella sp. isolated from children less than five years with diarrhoea attending 3 ... parasitic infections, as well as food intolerance, reaction to ..... Escherichia coil 0157:H7 as a model of entry of a new.

PESQUISA DE Salmonella spp. EM LEITE CRU, LEITE PASTEURIZADO TIPO C E QUEIJO "MINAS FRESCAL" COMERCIALIZADOS NO MUNICÍPIO DE PIRACICABA - SP

Directory of Open Access Journals (Sweden)

C.R. de ÁVILA

1996-01-01

Full Text Available Este trabalho foi conduzido para detectar a ocorrência de Salmonella spp. em leite cru, leite pasteurizado tipo C e queijo "Minas Frescal". Analisaram-se 19 amostras de cada produto de diferentes marcas comerciais, adquiridas em diversos estabelecimentos comerciais em Piracicaba, SP. Segundo o padrão de amostragem e métodos adotados, não foi detectada a presença de Salmonella spp. nas amostras analisadas. Durante este trabalho, foi possível observar que os 3 meios de isolamento utilizados: agar verde brilhante, agar bismuto sulfito e agar Salmonella-Shigella, apresentaram baixa seletividade, uma vez que permitiram o crescimento de bactérias não pertencentes ao gênero Salmonella. O mesmo ocorreu com os caldos tetrationato e selenito-cistina utilizados no enriquecimento seletivo.This work was carried out to detect the occurrence of Salmonella spp. in raw milk type C, pasteurized milk and "Minas Frescal" cheese. A total of 19 samples of each product of different commercial brands, from various retail stores in Piracicaba, SP, were analyzed. According to the pattern of sampling and methodology adopted, none of the samples was positive for Salmonella. During this work, it was possible to observe that the 3 isolation media utilized, brilliant green agar, bismuth sulfite agar and Salmonella-Shigella agar, presented low selectivity, since they allowed the growth of non Salmonella organisms. The same occurred with the selective enrichment: tetrathionate broth and selenite-cystine broth.

Development of an Aotus nancymaae Model for Shigella Vaccine Immunogenicity and Efficacy Studies

OpenAIRE

Gregory, Michael; Kaminski, Robert W.; Lugo-Roman, Luis A.; Galvez Carrillo, Hugo; Tilley, Drake Hamilton; Baldeviano, Christian; Simons, Mark P.; Reynolds, Nathanael D.; Ranallo, Ryan T.; Suvarnapunya, Akamol E.; Venkatesan, Malabi M.; Oaks, Edwin V.

2014-01-01

Several animal models exist to evaluate the immunogenicity and protective efficacy of candidate Shigella vaccines. The two most widely used nonprimate models for vaccine development include a murine pulmonary challenge model and a guinea pig keratoconjunctivitis model. Nonhuman primate models exhibit clinical features and gross and microscopic colonic lesions that mimic those induced in human shigellosis. Challenge models for enterotoxigenic Escherichia coli (ETEC) and Campylobacter spp. have...

Injury and recovery of salmonella, Escherichia coli 0157:H7 and Listeria Monocytogenes on cantaloupe rind surfaces after hyrdogren peroxide and minimal thermal treatment

Science.gov (United States)

Introduction: Produce surface structures vary and complicate decontamination treatments for reducing attached bacteria. Purpose: The objective of this study on survival and recovery of injured population of Salmonella, Escherichia coli O157:H7 and Listeria monocytogenes on cantaloupe rind surfaces...

Modification Of Carry-Blair Transport Media For Storage Salmonella typhi

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Yati Supriatin

2016-09-01

Full Text Available The aim of this study was to determine transport media modification as alternative media to replace Carry Blair. One type of transport media that often use to carry faeces specimens suspected to contain Salmonella typhi is Carry-Blair media. Studies have been conducted experimentally by storing Salmonella typhi on alternative transport media with Peptone composition, disodium Phosphate, Sodium chloride, Calcium chloride, which is made using a semi-solid and Carry-Blair as a control. Three variety of storage was done (0 hour,6 hours,9 hours at a temperature 4⁰-8⁰C and then Salmonella typhi was inoculated in Salmonella Shigella Agar using spread plate technique incubated during 24 hours at 37⁰C, counted the number of colonies by the plate count method using the colony counter. The results of ANOVA could be concluded that modification media could be use as alternative media replace Carry-Blair at 6 hours. Based on regression correlation test was assumed that the Salmonella typhi bacteria still life at less than 11 hours 54 minutes.

[Evaluation of the presence of hygienic and sanitary indicator microorganisms in food served in public schools in Porto Alegre, Brazil].

Science.gov (United States)

de Oliveira, Ana Beatriz Almeida; Capalonga, Roberta; Silveira, Joice Trindade; Tondo, Eduardo Cesar; Cardoso, Marisa Ribeiro de Itapema

2013-04-01

The objective of this study was to evaluate the presence of hygienic and sanitary indicator microorganisms in samples of food served in public schools in Porto Alegre. All the food served in the meal of the session visited was analyzed for Escherichia coli, coagulase-positive Staphylococcus, Salmonella sp. and Shigella sp. Of the total of 196 food products analyzed in 120 schools, 4 contained and Escherichia coli score above the permitted level, and 2 contained coagulase-positive Staphylococcus. Neither Shigella nor Salmonella genus were detected. In the majority of schools studied, it was found that food was of an adequate hygienic-sanitary standard. However, only municipal schools had the supervision of a technician responsible for school food. In the state schools, 60% had never been visited by a nutritionist and in these schools several procedures failed to comply with legal requirements. In most of the schools studied, the food served to students was within adequate standards, though the problems detected revealed the need for the implementation of Best Practices in the school environment.

DETECCIÓN DE Escherichia coli O157: H7 y Salmonella spp., EN CERDOS DEL DEPARTAMENTO DE CORDOBA

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Jaime Vargas

2004-07-01

Full Text Available E. coli O157:H7 y Salmonella spp., son bacterias de distribución mundial causantes de enfermedades intestinales queafectan tanto al hombre como a LOS animales. ESTE estudio tuvo como objetivo determinar la presencia y frecuenciade aparición de E. coli O157:H7 y Salmonella spp., en los diferentes sistemas de producción porcina que se empleanen el departamento de Córdoba. Se realizó un estudio de corte descriptivo prospectivo, con un muestreo al azar enlos sistemas de explotación porcina intensiva y extensiva. Se procesaron 500 muestras de materia fecal de porcinos,250 provenientes del sistema extensivo y 250 del sistema intensivo. Para la detección E. coli y Salmonella spp., sellevaron a cabo procedimientos estándares microbiológicos. Los resultados mostraron una frecuencia de aparición deSalmonella spp., del 1%, el 0.2% en el sistema intensivo y el 0.8% en el sistema extensivo; no se aisló Escherichia coliO157:H7. Los resultados de resistencia y sensibilidad a los antibióticos en las cepas aisladas de Salmonella spp.,mostraron una sensibilidad del 100% al trimetoprim sulfametozasol, a la amikacina, al ceftriaxona, a la ciprofloxacina,a la gentamicina y al aztreonam y un 20% a la ampicilina y al sulbactam. Se concluye que la frecuencia de apariciónde Salmonella spp., en muestras coprológicas porcinas es baja, y nula para E. coli O157:H7, sin embargo, se debemantener la vigilancia sobre estos patógenos, por lo que se recomienda continuar los estudios epidemiológicos.

Comparison of 61 Sequenced Escherichia coli Genomes

DEFF Research Database (Denmark)

Lukjancenko, Oksana; Wassenaar, T. M.; Ussery, David

2010-01-01

Escherichia coli is an important component of the biosphere and is an ideal model for studies of processes involved in bacterial genome evolution. Sixty-one publically available E. coli and Shigella spp. sequenced genomes are compared, using basic methods to produce phylogenetic and proteomics...

Electrochemical genosensing of Salmonella, Listeria and Escherichia coli on silica magnetic particles.

Science.gov (United States)

Liébana, Susana; Brandão, Delfina; Cortés, Pilar; Campoy, Susana; Alegret, Salvador; Pividori, María Isabel

2016-01-21

A magneto-genosensing approach for the detection of the three most common pathogenic bacteria in food safety, such as Salmonella, Listeria and Escherichia coli is presented. The methodology is based on the detection of the tagged amplified DNA obtained by single-tagging PCR with a set of specific primers for each pathogen, followed by electrochemical magneto-genosensing on silica magnetic particles. A set of primers were selected for the amplification of the invA (278 bp), prfA (217 bp) and eaeA (151 bp) being one of the primers for each set tagged with fluorescein, biotin and digoxigenin coding for Salmonella enterica, Listeria monocytogenes and E. coli, respectively. The single-tagged amplicons were then immobilized on silica MPs based on the nucleic acid-binding properties of silica particles in the presence of the chaotropic agent as guanidinium thiocyanate. The assessment of the silica MPs as a platform for electrochemical magneto-genosensing is described, including the main parameters to selectively attach longer dsDNA fragments instead of shorter ssDNA primers based on their negative charge density of the sugar-phosphate backbone. This approach resulted to be a promising detection tool with sensing features of rapidity and sensitivity very suitable to be implemented on DNA biosensors and microfluidic platforms. Copyright © 2015 Elsevier B.V. All rights reserved.

Quantitative Microbial Risk Assessment for Escherichia coli O157 : H7, Salmonella, and Listeria monocytogenes in Leafy Green Vegetables Consumed at Salad Bars

NARCIS (Netherlands)

Franz, E.; Tromp, S.O.; Rijgersberg, H.; Fels-Klerx, van der H.J.

2010-01-01

Fresh vegetables are increasingly recognized as a source of foodborne outbreaks in many parts of the world. The purpose of this study was to conduct a quantitative microbial risk assessment for Escherichia coli O157:H7, Salmonella, and Listeria monocytogenes infection from consumption of leafy green

Genomics of Escherichia and Shigella

Science.gov (United States)

Perna, Nicole T.

The laboratory workhorse Escherichia coli K-12 is among the most intensively studied living organisms on earth, and this single strain serves as the model system behind much of our understanding of prokaryotic molecular biology. Dense genome sequencing and recent insightful comparative analyses are making the species E. coli, as a whole, an emerging system for studying prokaryotic population genetics and the relationship between system-scale, or genome-scale, molecular evolution and complex traits like host range and pathogenic potential. Genomic perspective has revealed a coherent but dynamic species united by intraspecific gene flow via homologous lateral or horizontal transfer and differentiated by content flux mediated by acquisition of DNA segments from interspecies transfers.

Assessment of Consumer Exposure to Salmonella spp., Campylobacter spp., and Shiga Toxin-Producing Escherichia coli in Meat Products at Retail in the City of Sao Paulo, Brazil.

Science.gov (United States)

Ristori, Christiane Asturiano; Rowlands, Ruth Estela Gravato; Martins, Cecília Geraldes; Barbosa, Maria Luisa; Dos Santos, Luis Fernando; Jakabi, Miyoko; de Melo Franco, Bernadette Dora Gombossy

2017-08-01

Meat products may be vehicles of bacterial pathogens to humans, and Salmonella spp., Campylobacter spp., and Shiga toxin-producing Escherichia coli (STEC) are the most relevant. The aim of this study was to generate data on prevalence of these three pathogens in 552 samples of meat products (hot dogs, pork sausages, raw ground beef, and raw chicken legs) sold at retail in the city of Sao Paulo, Brazil. Salmonella spp. was detected in 5.8% (32/552) of samples, comprising pork sausages 62.5% (20/32) and chicken legs 37.5% (12/32). The counts of Salmonella spp. were low, ranging from Salmonella Typhimurium (28.1%), Salmonella I 4,[5],12:i:- (15.6%), Salmonella Enteritidis (12.5%), Salmonella Derby, and Salmonella Brandenburg (9.4%). Campylobacter spp. was detected in 33 samples (6.0%), comprising chicken legs (82%) and ground beef (18%). All samples were negative for STEC. These results suggest that meat products when subjected to inadequate cooking and/or cross-contamination with other products ready for consumption can lead to occurrence of outbreaks, highlighting the risks associated with them.

Glutathione: an intracellular and extracellular protective agent in Salmonella typhimurium and Escherichia coli

International Nuclear Information System (INIS)

Owens, R.A.

1986-01-01

Levels of glutathione, were measured in several aerobically grown strains of Salmonella typhimurium and Escherichia coli. External accumulation of GSH was inhibited by 30 mM NaN 3 . Thus, GSH export may be energy dependent. Greater than 50% of the glutathione detected in the media was in the reduced form. Since the oxidized glutathione in the media could be accounted for by oxidation during aerobic incubation as well as in sample processing, the glutathione was predominantly exported in the reduced form. Extracellular glutathione was detected in log phase cultures of 2 out of 2 E. coli strains and 6 of 8 Salmonella strains tested. Two-dimensional paper chromatography of supernatants from cultures labelled with Na 2 35 SO 4 confirmed the presence of GSH and revealed five other sulfur-containing compounds in the media of Salmonella and E. coli cultures. Since media from cultures of an E. coli GSH - strain contained compounds with identical R/sub f/'s, the five unidentified compounds were not derivatives of GSH. The addition of 26 μM GSH to cultures of TA1534 partially protected the bacteria from the toxic effects of 54 μM N-methyl-N'-nitro-N-nitrosoguanidine (MNNG). When MNNG was preincubated with equimolar GSH, the mutagenicity of the MNNG was neutralized. The addition of micromolar GSH to cultures and E. coli GSH - strain protected the cells from growth inhibition by micromolar concentrations of mercuric chloride, silver nitrate, cisplatin, cadmium chloride, and iodoacetamide. The data presented demonstrate that micromolar concentrations of external GSH can significantly shorten the recovery time of cells after exposure to toxic agents in the environment

Inactivation of Listeria monocytogenes, Escherichia coli O157:H7, and Salmonella typhimurium with compounds available in households.

Science.gov (United States)

Yang, Hua; Kendall, Patricia A; Medeiros, Lydia; Sofos, John N

2009-06-01

Solutions of selected household products were tested for their effectiveness against Listeria monocytogenes, Escherichia coli O157:H7, and Salmonella Typhimurium. Hydrogen peroxide (1.5 and 3%), vinegar (2.5 and 5% acetic acid), baking soda (11, 33, and 50% sodium bicarbonate), household bleach (0.0314, 0.0933, and 0.670% sodium hypochlorite), 5% acetic acid (prepared from glacial acetic acid), and 5% citric acid solutions were tested against the three pathogens individually (five-strain composites of each, 10(8) CFU/ml) by using a modified AOAC International suspension test at initial temperatures of 25 and 55degrees C for 1 and 10 min. All bleach solutions (pH 8.36 to 10.14) produced a >5-log reduction of all pathogens tested after 1 min at 25 degrees C, whereas all baking soda solutions (pH 7.32 to 7.55) were ineffective (5-log reduction of both Salmonella Typhimurium and E. coli O157:H7, whereas undiluted vinegar (pH 2.58) had a similar effect only against Salmonella Typhimurium. Compared with 1 min at 25 degrees C, greater reductions of L. monocytogenes (P 3% hydrogen peroxide > undiluted vinegar and 5% acetic acid > 5% citric acid > baking soda (50% sodium bicarbonate). The sensitivity of the tested pathogens to all tested household compounds followed the sequence of Salmonella Typhimurium > E. coli O157: H7 > L. monocytogenes.

Potential public health significance of faecal contamination and multidrug-resistant Escherichia coli and Salmonella serotypes in a lake in India.

Science.gov (United States)

Abhirosh, C; Sherin, V; Thomas, A P; Hatha, A A M; Mazumder, A

2011-06-01

To assess the prevalence of faecal coliform bacteria and multiple drug resistance among Escherichia coli and Salmonella serotypes from Vembanadu Lake. Systematic microbiological testing. Monthly collection of water samples were made from ten stations on the southern and northern parts of a salt water regulator constructed in Vembanadu Lake in order to prevent incursion of seawater during certain periods of the year. Density of faecal colifrom bacteria was estimated. E. coli and Salmonella were isolated and their different serotypes were identified. Antibiotic resistance analysis of E. coli and Salmonella serotypes was done and the MAR index of individual isolates was calculated. Density of faecal coliform bacteria ranged from mean MPN value 2900 -7100/100ml. Results showed multiple drug resistance pattern among the bacterial isolates. E. coli showed more than 50% resistance to amickacin, oxytetracycline, streptomycin, tetracycline and kanamycin while Salmonella showed high resistance to oxytetracycline, streptomycin, tetracycline and ampicillin. The MAR indexing of the isolates showed that they have originated from high risk source such as humans, poultry and dairy cows. The high density of faecal coliform bacteria and prevalence of multi drug resistant E. coli and Salmonella serotypes in the lake may pose severe public health risk through related water borne and food borne outbreaks. Copyright © 2011 The Royal Society for Public Health. Published by Elsevier Ltd. All rights reserved.

Antimicrobial activity against Shigella sonnei and probiotic properties of wild lactobacilli from fermented food.

Science.gov (United States)

Zhang, Yingchun; Zhang, Lanwei; Du, Ming; Yi, Huaxi; Guo, Chunfeng; Tuo, Yanfeng; Han, Xue; Li, Jingyan; Zhang, Lili; Yang, Lin

2011-12-20

Four lactobacilli strains (Lactobacillus paracasei subp. paracasei M5-L, Lactobacillus rhamnosus J10-L, Lactobacillus casei Q8-L and L. rhamnosus GG (LGG), were systematically assessed for the production of antimicrobial substances active towards Shigella sonnei, Escherichia coli and Salmonella typhimurium. Agar-well assay showed that the four lactobacilli strains displayed strong antibacterial activity towards S. sonnei. The nature of antimicrobial substances was also investigated and shown to be dependent on the production of organic acids, in particular the lactic acid. Time-kill assay showed that the viability of the S. sonnei was decreased by 2.7-3.6logCFU/ml after contact with CFCS (cell-free culture supernatants) of four lactobacilli for 2h, which confirmed the result of the agar-well assay. Further analysis of the organic acid composition in the CFCS revealed that the content of lactic acid range from 227 to 293mM. In addition, the aggregations properties, adherence properties and tolerance to simulated gastrointestinal conditions were also investigated in vitro tests. The result suggested that the M5-L, J10-L and Q8-L strains possess desirable antimicrobial activity towards S. sonnei and probiotic properties as LGG and could be potentially used as novel probiotic strains in the food industry. Copyright © 2011. Published by Elsevier GmbH.

Impact of storm runoff on Salmonella and Escherichia coli prevalence in irrigation ponds of fresh produce farms in southern Georgia.

Science.gov (United States)

Harris, C S; Tertuliano, M; Rajeev, S; Vellidis, G; Levy, K

2018-03-01

To examine Salmonella and Escherichia coli in storm runoff and irrigation ponds used by fresh produce growers, and compare Salmonella serovars with those found in cases of human salmonellosis. We collected water before and after rain events at two irrigation ponds on farms in southern Georgia, USA, and collected storm runoff/storm flow within the contributing watershed of each pond. Salmonella and E. coli concentrations were higher in ponds after rain events by an average of 0·46 (P storm runoff from fields and forests were not significantly higher than in ponds before rain events, but concentrations in storm flow from streams and ditches were higher by an average of 1·22 log 10 MPN per 100 ml (P storm runoff/storm flow and ponds. Seven of the serovars, including five of the shared serovars, were present in cases of human illness in the study region in the same year. However, several serovars most commonly associated with human illness in the study region (e.g. Javiana, Enteritidis, and Montevideo) were not found in any water samples. Salmonella and E. coli concentrations in irrigation ponds were higher, on average, after rain events, but concentrations of Salmonella were low, and the ponds met FDA water quality standards based on E. coli. Some similarities and notable differences were found between Salmonella serovars in water samples and in cases of human illness. This study directly examined storm runoff/storm flow into irrigation ponds and quantified increases in Salmonella and E. coli following rain events, with potential implications for irrigation pond management as well as human health. © 2018 The Society for Applied Microbiology.

Small-Molecule Inhibitor of the Shigella flexneri Master Virulence Regulator VirF

Science.gov (United States)

Koppolu, Veerendra; Osaka, Ichie; Skredenske, Jeff M.; Kettle, Bria; Hefty, P. Scott; Li, Jiaqin

2013-01-01

VirF is an AraC family transcriptional activator that is required for the expression of virulence genes associated with invasion and cell-to-cell spread by Shigella flexneri, including multiple components of the type three secretion system (T3SS) machinery and effectors. We tested a small-molecule compound, SE-1 (formerly designated OSSL_051168), which we had identified as an effective inhibitor of the AraC family proteins RhaS and RhaR, for its ability to inhibit VirF. Cell-based reporter gene assays with Escherichia coli and Shigella, as well as in vitro DNA binding assays with purified VirF, demonstrated that SE-1 inhibited DNA binding and transcription activation (likely by blocking DNA binding) by VirF. Analysis of mRNA levels using real-time quantitative reverse transcription-PCR (qRT-PCR) further demonstrated that SE-1 reduced the expression of the VirF-dependent virulence genes icsA, virB, icsB, and ipaB in Shigella. We also performed eukaryotic cell invasion assays and found that SE-1 reduced invasion by Shigella. The effect of SE-1 on invasion required preincubation of Shigella with SE-1, in agreement with the hypothesis that SE-1 inhibited the expression of VirF-activated genes required for the formation of the T3SS apparatus and invasion. We found that the same concentrations of SE-1 had no detectable effects on the growth or metabolism of the bacterial cells or the eukaryotic host cells, respectively, indicating that the inhibition of invasion was not due to general toxicity. Overall, SE-1 appears to inhibit transcription activation by VirF, exhibits selectivity toward AraC family proteins, and has the potential to be developed into a novel antibacterial agent. PMID:24002059

Prediction of Salmonella carcass contamination by a comparative quantitative analysis of E. coli and Salmonella during pig slaughter

DEFF Research Database (Denmark)

Nauta, Maarten; Barfod, Kristen; Hald, Tine

2013-01-01

Salmonella concentrations. It is concluded that the faecal carriage of Salmonella together with the faecal contamination of carcasses, as predicted from E. coli data in the animal faeces and hygiene performance of the slaughterhouse, is not sufficient to explain carcass contamination with Salmonella. Our...... extensive data set showed that other factors than the observed faecal carriage of Salmonella by the individual animals brought to slaughter, play a more important role in the Salmonella carcass contamination of pork.......Faecal contamination of carcasses in the slaughterhouse is generally considered to be the source of Salmonella on pork. In this study the hygiene indicator Escherichia coli is used to quantify faecal contamination of carcasses and it is hypothesized that it can be used to predict the quantitative...

Prevalence of beta-lactamases among ampicillin-resistant Escherichia coli and Salmonella isolated from food animals in Denmark

DEFF Research Database (Denmark)

Olesen, Inger; Hasman, Henrik; Aarestrup, Frank Møller

2004-01-01

The genetic background for beta-lactamase-mediated resistance to beta-lactam antibiotics was examined by PCR and sequencing in 160 ampicillin-resistant isolates (109 Escherichia coli and 51 Salmonella) obtained from healthy and diseased food animals in Denmark. Sequencing revealed three different...... leading to increased production of the AmpC beta-lactamase were demonstrated in 11 cefoxitin-resistant or intermediate E. coli isolates. Nine of these isolates did not contain any bla(TEM) genes, whereas the remaining two did. No genes encoding SHV or extended-spectrum beta-lactamases were detected. Two...

International collaborative study on the occurrence of plasmid-mediated quinolone resistance in Salmonella enterica and Escherichia coli isolated from animals, humans, food and the environment in 13 European countries

DEFF Research Database (Denmark)

Veldman, Kees; Cavaco, Lina; Mevius, Dik

2011-01-01

OBJECTIVES: This study was initiated to collect retrospective information on the occurrence of plasmid-mediated quinolone resistance (PMQR) in Salmonella enterica and Escherichia coli isolates in Europe and to identify the responsible genes. METHODS: Databases of national reference laboratories...... containing MIC values for Salmonella and E. coli isolated between 1994 and 2009 in animals, humans, food and the environment from 13 European countries were screened for isolates exhibiting a defined quinolone resistance phenotype, i.e. reduced susceptibility to fluoroquinolones and nalidixic acid. PCR...... isolate. No qnrC or qepA genes were detected in either Salmonella or E. coli. CONCLUSIONS: This study shows the occurrence and dissemination of PMQR genes in Salmonella and E. coli in Europe with a defined quinolone resistance phenotype. We also report the first detection of qnrD in Salmonella collected...

Technology Assessment and Strategy for Development of a Rapid Field Water Microbiology Test Kit

Science.gov (United States)

1991-09-01

lientamoeba, Enteromonas, and Chil)mastix (Heyneman 1987), (2) SARCODINA, which are typically ameboid, intluding Entamoeba , Endolimax, Iodamoeia, Naeleril...infective dose in man. [From Drasar and Barrow (1985)]. Organism Infective Dose Escherichia coli 106-108 Salmonella sp . 104_108’ Shigella sp . 10-200...intestine. Worldwide tropics and North America Ancylostoma braziliense (N) Subcutaneous Worldwide Toxocara sp . (N) Lung, liver, eye, Worldwide brain

Campylobacter, Salmonella, Listeria monocytogenes, verotoxigenic Escherichia coli, and Escherichia coli prevalence, enumeration, and subtypes on retail chicken breasts with and without skin.

Science.gov (United States)

Cook, Angela; Odumeru, Joseph; Lee, Susan; Pollari, Frank

2012-01-01

This study examined the prevalence, counts, and subtypes of Campylobacter, Salmonella, Listeria monocytogenes, verotoxigenic Escherichia coli (VTEC), and E. coli on raw retail chicken breast with the skin on versus the skin off. From January to December 2007, 187 raw skin-on chicken breasts and 131 skin-off chicken breasts were collected from randomly selected retail grocery stores in the Region of Waterloo, Ontario, Canada. Campylobacter isolates were recovered from a higher proportion of the skin-off chicken breasts, 55 (42%) of 131, than of the skin-on chicken breasts tested, 55 (29%) of 187 (P = 0.023). There was no difference in the proportion of Salmonella isolates recovered from the two meat types (P = 0.715): 40 (31%) of 131 skin-off chicken breasts versus 61 (33%) of 187 skin-on chicken breasts. L. monocytogenes isolates were recovered from a statistically lower proportion of the skin-off chicken breasts, 15 (15%) of 99, than of the skin-on chicken breasts, 64 (34%) of 187 (P = 0.001). There was no difference in the proportion of E. coli isolates recovered from the skin-off chicken breasts, 33 (33%) of 99, than from the skin-on chicken breasts, 77 (41%) of 187 (P = 0.204). VTEC was detected on a single skin-off chicken breast. Campylobacter jejuni was the most frequent species isolated on both types of chicken meat: skin-on, 48 (87%) of 55, and skin-off, 51 (94%) of 54. Salmonella serotypes Kentucky and Heidelberg and L. monocytogenes serotype 1/2a were the most frequently detected serotypes from both skin-off and skin-on chicken breasts. Although there appeared to be a trend toward higher enumeration values of these pathogens and E. coli on the skin-on chicken, the differences did not exceed 1 log. This study suggested that skin-off chicken breast may represent a higher risk of consumer exposure to Campylobacter, a similar risk for Salmonella, VTEC, and E. coli, and a lower risk for L. monocytogenes than skin-on chicken breast.

Hemagglutinin Typing as an Aid in Identification of Biochemically Atypical Escherichia coli Strains

OpenAIRE

Crichton, Pamela B.; Ip, S. M.; Old, D. C.

1981-01-01

Tests for the presence of mannose-sensitive and mannose-resistant, eluting hemagglutinins and fimbriae were helpful in indicating whether biochemically atypical strains of the tribe Escherichieae might be escherichiae or shigellae.

Phylogenetic relationships of Salmonella based on rRNA sequences

DEFF Research Database (Denmark)

Christensen, H.; Nordentoft, Steen; Olsen, J.E.

1998-01-01

separated by 16S rRNA analysis and found to be closely related to the Escherichia coli and Shigella complex by both 16S and 23S rRNA analyses. The diphasic serotypes S. enterica subspp. I and VI were separated from the monophasic serotypes subspp. IIIa and IV, including S. bongori, by 23S rRNA sequence...

Hemagglutinin Typing as an Aid in Identification of Biochemically Atypical Escherichia coli Strains

Science.gov (United States)

Crichton, Pamela B.; Ip, S. M.; Old, D. C.

1981-01-01

Tests for the presence of mannose-sensitive and mannose-resistant, eluting hemagglutinins and fimbriae were helpful in indicating whether biochemically atypical strains of the tribe Escherichieae might be escherichiae or shigellae. PMID:7334072

Characterization of epidemic IncI1-Iγ plasmids harboring ambler class A and C genes in Escherichia coli and Salmonella enterica from animals and humans

NARCIS (Netherlands)

Smith, Hilde; Bossers, Alex; Harders, Frank; Wu, Guanghui; Woodford, Neil; Schwarz, Stefan; Guerra, Beatriz; Rodríguez, Irene; van Essen-Zandbergen, Alieda; Brouwer, Michael; Mevius, Dik

The aim of the study was to identify the plasmid-encoded factors contributing to the emergence and spread of epidemic IncI1-Iγ plasmids obtained from Escherichia coli and Salmonella enterica isolates from animal and human reservoirs. For this, 251 IncI1-Iγ plasmids carrying various extended-spectrum

Shigella

Science.gov (United States)

Marteyn, Benoit; Gazi, Anastasia; Sansonetti, Philippe

2012-01-01

Much is known about the molecular effectors of pathogenicity of gram-negative enteric pathogens, among which Shigella can be considered a model. This is due to its capacity to recapitulate the multiple steps required for a pathogenic microbe to survive close to its mucosal target, colonize and then invade its epithelial surface, cause its inflammatory destruction and simultaneously regulate the extent of the elicited innate response to likely survive the encounter and achieve successful subsequent transmission. These various steps of the infectious process represent an array of successive environmental conditions to which the bacteria need to successfully adapt. These conditions represent the selective pressure that triggered the “arms race” in which Shigella acquired the genetic and molecular effectors of its pathogenic armory, including the regulatory hierarchies that regulate the expression and function of these effectors. They also represent cues through which Shigella achieves the temporo-spatial expression and regulation of its virulence effectors. The role of such environmental cues has recently become obvious in the case of the major virulence effector of Shigella, the type three secretion system (T3SS) and its dedicated secreted virulence effectors. It needs to be better defined for other major virulence components such as the LPS and peptidoglycan which are used as examples here, in addition to the T3SS as models of regulation as it relates to the assembly and functional regulation of complex macromolecular systems of the bacterial surface. This review also stresses the need to better define what the true and relevant environmental conditions can be at the various steps of the progression of infection. The “identity” of the pathogen differs depending whether it is cultivated under in vitro or in vivo conditions. Moreover, this “identity” may quickly change during its progression into the infected tissue. Novel concepts and relevant tools are

Distribution of extended-spectrum cephalosporin resistance determinants in Salmonella enterica and Escherichia coli isolated from broilers in southern Japan.

Science.gov (United States)

Shahada, F; Chuma, T; Kosugi, G; Kusumoto, M; Iwata, T; Akiba, M

2013-06-01

This study was conducted to investigate the distribution and diversity of extended-spectrum cephalosporin (ESC) resistance determinants in Salmonella enterica and Escherichia coli obtained from the same cecal samples and to provide evidence of transmission of the resistance determinants among these bacteria in broiler farms in southern Japan. Salmonella enterica and E. coli were characterized by serotyping and multilocus sequence typing, respectively. An antimicrobial susceptibility test, plasmid analysis, and identification and localization of resistance genes were performed to determine the relatedness of ESC resistance determinants among the isolates. Of 48 flocks examined, 14 had S. enterica. In total, 57 S. enterica isolates were obtained, 45 of which showed ESC resistance. Extended-spectrum cephalosporin-resistant E. coli were also obtained from all of these ESC-resistant Salmonella-positive samples. β-Lactamase genes, blaTEM-52 (38 isolates), blaCTX-M-14 (1 isolate), and blaCMY-2 (6 isolates), were carried by conjugative untypable or IncP plasmids detected in the S. enterica serovars Infantis and Manhattan. The β-lactamase genes blaCTX-M-14 (3 isolates), blaCTX-M-15 (3 isolates), blaSHV-2 (1 isolate), blaSHV-12 (2 isolates), and blaCMY-2 (32 isolates) associated with IncI1-Iγ, IncFIB, IncFIC, IncK, IncB/O, and IncY plasmids were detected in E. coli co-isolates. Restriction mapping revealed similar plasmids in Salmonella Infantis and Salmonella Manhattan and in different sequence types of E. coli. Intraspecies transmission of plasmids was suggested within S. enterica and E. coli populations, whereas interspecies transmission was not observed. This study highlights the importance of plasmids as carriers of ESC resistance determinants.

Relationship between drug resistance and the clustered, regularly interspaced, short, palindromic repeat-associated protein genes cas1 and cas2 in Shigella from giant panda dung.

Science.gov (United States)

Ren, Lu; Deng, Lin-Hua; Zhang, Ri-Peng; Wang, Cheng-Dong; Li, De-Sheng; Xi, Li-Xin; Chen, Zhen-Rong; Yang, Rui; Huang, Jie; Zeng, Yang-Ru; Wu, Hong-Lin; Cao, San-Jie; Wu, Rui; Huang, Yong; Yan, Qi-Gui

2017-02-01

To detect drug resistance in Shigella obtained from the dung of the giant panda, explore the factors leading to drug resistance in Shigella, understand the characteristics of clustered, regularly interspaced, short, palindromic repeats (CRISPR), and assess the relationship between CRISPR and drug resistance. We collected fresh feces from 27 healthy giant pandas in the Giant Panda Conservation base (Wolong, China). We identified the strains of Shigella in the samples by using nucleotide sequence analysis. Further, the Kirby-Bauer paper method was used to determine drug sensitivity of the Shigella strains. CRISPR-associated protein genes cas1 and cas2 in Shigella were detected by polymerase chain reaction (PCR), and the PCR products were sequenced and compared. We isolated and identified 17 strains of Shigella from 27 samples, including 14 strains of Shigella flexneri, 2 strains of Shigella sonnei, and 1 strain of Shigella dysenteriae. Further, drug resistance to cefazolin, imipenem, and amoxicillin-clavulanic acid was identified as a serious problem, as multidrug-resistant strains were detected. Further, cas1 and cas2 showed different degrees of point mutations. The CRISPR system widely exists in Shigella and shares homology with that in Escherichia coli. The cas1 and cas 2 mutations contribute to the different levels of resistance. Point mutations at sites 3176455, 3176590, and 3176465 in cas1 (a); sites 3176989, 3176992, and 3176995 in cas1 (b); sites 3176156 and 3176236 in cas2 may affect the resistance of bacteria, cause emergence of multidrug resistance, and increase the types of drug resistance.

Characteristics of Clusters of Salmonella and Escherichia coli O157 Detected by Pulsed-Field Gel Electrophoresis that Predict Identification of Outbreaks.

Science.gov (United States)

Jones, Timothy F; Sashti, Nupur; Ingram, Amanda; Phan, Quyen; Booth, Hillary; Rounds, Joshua; Nicholson, Cyndy S; Cosgrove, Shaun; Crocker, Kia; Gould, L Hannah

2016-12-01

Molecular subtyping of pathogens is critical for foodborne disease outbreak detection and investigation. Many clusters initially identified by pulsed-field gel electrophoresis (PFGE) are not confirmed as point-source outbreaks. We evaluated characteristics of clusters that can help prioritize investigations to maximize effective use of limited resources. A multiagency collaboration (FoodNet) collected data on Salmonella and Escherichia coli O157 clusters for 3 years. Cluster size, timing, extent, and nature of epidemiologic investigations were analyzed to determine associations with whether the cluster was identified as a confirmed outbreak. During the 3-year study period, 948 PFGE clusters were identified; 849 (90%) were Salmonella and 99 (10%) were E. coli O157. Of those, 192 (20%) were ultimately identified as outbreaks (154 [18%] of Salmonella and 38 [38%] of E. coli O157 clusters). Successful investigation was significantly associated with larger cluster size, more rapid submission of isolates (e.g., for Salmonella, 6 days for outbreaks vs. 8 days for nonoutbreaks) and PFGE result reporting to investigators (16 days vs. 29 days, respectively), and performance of analytic studies (completed in 33% of Salmonella outbreaks vs. 1% of nonoutbreaks) and environmental investigations (40% and 1%, respectively). Intervals between first and second cases in a cluster did not differ significantly between outbreaks and nonoutbreaks. Molecular subtyping of pathogens is a rapidly advancing technology, and successfully identifying outbreaks will vary by pathogen and methods used. Understanding criteria for successfully investigating outbreaks is critical for efficiently using limited resources.

Preparation, crystallization and preliminary X-ray crystallographic studies of diadenosine tetraphosphate hydrolase from Shigella flexneri 2a

Energy Technology Data Exchange (ETDEWEB)

Hu, Wenxin; Wang, Qihai; Bi, Ruchang, E-mail: rcbi@sun5.ibp.ac.cn [Institute of Biophysics, Chinese Academy of Sciences, 15 Datun Road, Chaoyang District, Beijing 100101 (China)

2005-12-01

The 31.3 kDa Ap{sub 4}A hydrolase from Shigella flexneri 2a has been cloned, expressed and purified using an Escherichia coli expression system. Crystals of Ap{sub 4}A hydrolase have been obtained by the hanging-drop technique at 291 K using PEG 550 MME as precipitant. Diadenosine tetraphosphate (Ap{sub 4}A) hydrolase (EC 3.6.1.41) hydrolyzes Ap{sub 4}A symmetrically in prokaryotes. It plays a potential role in organisms by regulating the concentration of Ap{sub 4}A in vivo. To date, no three-dimensional structures of proteins with significant sequence homology to this protein have been determined. The 31.3 kDa Ap{sub 4}A hydrolase from Shigella flexneri 2a has been cloned, expressed and purified using an Escherichia coli expression system. Crystals of Ap{sub 4}A hydrolase have been obtained by the hanging-drop technique at 291 K using PEG 550 MME as precipitant. Ap{sub 4}A hydrolase crystals diffract X-rays to 3.26 Å and belong to space group P2{sub 1}, with unit-cell parameters a = 118.9, b = 54.6, c = 128.5 Å, β = 95.7°.

Radiation inactivation of Salmonella panama and Escherichia coli K 12 present on deep-frozen broiler carcasses

International Nuclear Information System (INIS)

Mulder, R.W.A.W.

1976-01-01

Low doses of ionizing radiation have been used to extend the shelf life of refrigerated poultry carcasses and to reduce the numbers of Salmonellae present. This report gives results of experiments on irradiation of deep-frozen poultry carcasses which were, before freezing, artificially contaminated with Salmonella panama and with a nalidixic acid-resistant Escherichia coli K 12. The D-values (decimal reduction) obtained with the inoculated carcasses were compared with D-values obtained with carcasses which were slaughtered in the normal way. The D-values for S.panama and for E.coli K 12 were 64.9 krad and 55.9 krad in the dripwater. Under commercial conditions approximately 100 krad were required for one decimal reduction of the Enterobacteriaceae present. The D-values estimated on the skin were higher for S.panama than for E.coli K 12 (128.6 krad vs 57.6 krad). If it is assumed that 1 positive carcass in 10,000 is allowed, the deep-frozen carcasses should be irradiated with doses of at least 700 krad to be sure of the absence of the tested S.panama strain. (orig.) [de

Radiation inactivation of Salmonella panama and Escherichia coli K 12 present on deep-frozen broiler carcasses

Energy Technology Data Exchange (ETDEWEB)

Mulder, R W.A.W. [Spelderholt Inst. for Poultry Research, Beekbergen (Netherlands). Processing Dept.

1976-01-01

Low doses of ionizing radiation have been used to extend the shelf life of refrigerated poultry carcasses and to reduce the numbers of Salmonellae present. This report gives results of experiments on irradiation of deep-frozen poultry carcasses which were, before freezing, artificially contaminated with Salmonella panama and with a nalidixic acid-resistant Escherichia coli K 12. The D-values (decimal reduction) obtained with the inoculated carcasses were compared with D-values obtained with carcasses which were slaughtered in the normal way. The D-values for S.panama and for E.coli K 12 were 64.9 krad and 55.9 krad in the dripwater. Under commercial conditions approximately 100 krad were required for one decimal reduction of the Enterobacteriaceae present. The D-values estimated on the skin were higher for S.panama than for E.coli K 12 (128.6 krad vs 57.6 krad). If it is assumed that 1 positive carcass in 10,000 is allowed, the deep-frozen carcasses should be irradiated with doses of at least 700 krad to be sure of the absence of the tested S.panama strain.

Seasonal stability of Cladophora-associated Salmonella in Lake Michigan watersheds

Science.gov (United States)

Byappanahalli, M.N.; Sawdey, R.; Ishii, S.; Shively, D.A.; Ferguson, J.A.; Whitman, R.L.; Sadowsky, M.J.

2009-01-01

The bacterial pathogens Shigella, Salmonella, Campylobacter, and shiga toxin-producing E. coli (STEC) were recently found to be associated with Cladophora growing in southern Lake Michigan. Preliminary results indicated that the Salmonella strains associated with Cladophora were genetically identical to each other. However, because of the small sample size (n = 37 isolates) and a lack of information on spatial-temporal relationships, the nature of the association between Cladophora and Salmonella remained speculative. In this study, we investigated the population structure and genetic relatedness of a large number of Cladophora-borne Salmonella isolates from Lake Michigan (n = 133), as well as those isolated from stream and lake water (n = 31), aquatic plants (n = 8), and beach sands and sediments (n = 8) from adjacent watersheds. Salmonella isolates were collected during 2005-2007 between May and August from Lake Michigan beachsheds in Wisconsin, Illinois, and Indiana. The genetic relatedness of Salmonella isolates was examined by using the horizontal, fluorophore-enhanced rep-PCR (HFERP) DNA fingerprinting technique. While the Salmonella isolates associated with Cladophora exhibited a high degree of genetic relatedness (???92% similarity), the isolates were not all genetically identical. Spatial and temporal relationships were evident in the populations examined, with tight clustering of the isolates both by year and location. These findings suggest that the relationship between Salmonella and Cladophora is likely casual and is related to input sources (e.g. wastewater, runoff, birds) and the predominant Salmonella genotype surviving in the environment during a given season. Our studies indicate that Cladophora is likely an important reservoir for Salmonella and other enteric bacterial pathogens in Lake Michigan beachsheds, which in turn may influence nearshore water quality. ?? 2008 Elsevier Ltd.

Seasonal stability of Cladophora-associated Salmonella in Lake Michigan watersheds.

Science.gov (United States)

Byappanahalli, Muruleedhara N; Sawdey, Richard; Ishii, Satoshi; Shively, Dawn A; Ferguson, John A; Whitman, Richard L; Sadowsky, Michael J

2009-02-01

The bacterial pathogens Shigella, Salmonella, Campylobacter, and shiga toxin-producing E. coli (STEC) were recently found to be associated with Cladophora growing in southern Lake Michigan. Preliminary results indicated that the Salmonella strains associated with Cladophora were genetically identical to each other. However, because of the small sample size (n=37 isolates) and a lack of information on spatial-temporal relationships, the nature of the association between Cladophora and Salmonella remained speculative. In this study, we investigated the population structure and genetic relatedness of a large number of Cladophora-borne Salmonella isolates from Lake Michigan (n=133), as well as those isolated from stream and lake water (n=31), aquatic plants (n=8), and beach sands and sediments (n=8) from adjacent watersheds. Salmonella isolates were collected during 2005-2007 between May and August from Lake Michigan beachsheds in Wisconsin, Illinois, and Indiana. The genetic relatedness of Salmonella isolates was examined by using the horizontal, fluorophore-enhanced rep-PCR (HFERP) DNA fingerprinting technique. While the Salmonella isolates associated with Cladophora exhibited a high degree of genetic relatedness (>or=92% similarity), the isolates were not all genetically identical. Spatial and temporal relationships were evident in the populations examined, with tight clustering of the isolates both by year and location. These findings suggest that the relationship between Salmonella and Cladophora is likely casual and is related to input sources (e.g. wastewater, runoff, birds) and the predominant Salmonella genotype surviving in the environment during a given season. Our studies indicate that Cladophora is likely an important reservoir for Salmonella and other enteric bacterial pathogens in Lake Michigan beachsheds, which in turn may influence nearshore water quality.

Prevalence of Escherichia coli O157:H7 and Salmonella in camels, cattle, goats, and sheep harvested for meat in Riyadh.

Science.gov (United States)

Bosilevac, Joseph M; Gassem, Mustafa A; Al Sheddy, Ibraheem A; Almaiman, Salah A; Al-Mohizea, Ibrahim S; Alowaimer, Abdullah; Koohmaraie, Mohammad

2015-01-01

Escherichia coli O157:H7 and Salmonella are significant foodborne pathogens that can be found in the feces and on the hides of meat animals. When hides are removed during the harvest process, the carcass and subsequent meat products can become contaminated. Camels, cattle, sheep, and goats are harvested for meat in Riyadh, Saudi Arabia. The prevalence of E. coli O157:H7 and Salmonella are unknown in these animals, and it is assumed that if the animals carry the pathogens in their feces or on their hides, meat products are likely to become contaminated. To this end, a minimum of 206 samples each from hides and feces of camels, cattle, goats, and sheep were collected over the course of 8 months and tested for E. coli O157:H7 and Salmonella. It was found that E. coli O157:H7 was present in feces (10.7, 1.4, 2.4, and 2.4%) and on hides (17.9, 8.2, 2.9, and 9.2%) of cattle, goats, camels, and sheep, respectively. The prevalence of Salmonella was 11.2, 13.5, 23.2, and 18.8% in feces and 80.2, 51.2 67.6, and 60.2% on hides of cattle, goats, camels, and sheep, respectively. The prevalence of E coli O157:H7 was nearly zero in all samples collected in June and July, while Salmonella did not exhibit any seasonal variation. These results constitute the first comprehensive study of E. coli O157:H7 and Salmonella prevalence in Saudi Arabian meat animals at harvest.

Catecholamines and in vitro growth of pathogenic bacteria: enhancement of growth varies greatly among bacterial species

Science.gov (United States)

Belay, Tesfaye; Aviles, Hernan; Vance, Monique; Fountain, Kimberly; Sonnenfeld, Gerald

2003-01-01

The purpose of this study was to examine the effects of catecholamines on in vitro growth of a range of bacterial species, including anaerobes. Bacteria tested included: Porphyromonas gingivalis, Bacteriodes fragilis, Shigella boydii, Shigella sonnie, Enterobacter Sp, and Salmonella choleraesuis. The results of the current study indicated that supplementation of bacterial cultures in minimal medium with norepinephrine or epinephrine did not result in increased growth of bacteria. Positive controls involving treatment of Escherichia coli with catecholamines did result in increased growth of that bacterial species. The results of the present study extend previous observations that showed differential capability of catecholamines to enhance bacterial growth in vitro.

Efficacy of integrated treatment of UV light and low dose gamma irradiation on Escherichia coli O157:H7 and Salmonella enterica on grape tomatoes

Science.gov (United States)

Efficacy of integrated treatment of UVC and low dose Gamma irradiation to inactivate mixed Strains of Escherichia coli O157:H7 and Salmonella enterica inoculated on whole Grape tomatoes was evaluated. A mixed bacterial cocktail composed of a three strain mixture of E. coli O157:H7 (C9490, E02128 an...

DETEKSI CEMARAN BAKTERI KOLIFORM DAN Salmonella sp. PADA TEMPE YANG DIKEMAS DAUN PISANG DI DAERAH SALATIGA

Directory of Open Access Journals (Sweden)

Khanifa Nurul Khaq

2017-01-01

Full Text Available ABSTRACT Banana leaves-wrapped tempeh is a traditional food as a conventional base of vegetable protein source. Fermented soybeans into tempeh assisted by fungi kind Rhizopusoligosporus, Rhizopusoryzae and Rhizopusstolonifer. Hygiene is very important in the process of fermentation as it will affect the end result of products. Packaging materials can serve as a protective product, but related to the materials used can be the sources of microbial contaminants in food packaging.For small-scale Tempeh industry in Salatiga area commonly use banana leaves to wrap tempeh.The absence of leaves-wrapped tempeh quality standard made the manufacturers ignore the tempeh processing standard quality. The purpose of this study is to obtain data on the number of coliforms contamination and the presence or absence of Salmonella sp. contamination on banana leaves-wrapped soybean tempeh from manufacturer of production 5-10 kg per day in District of Sidorejo and Tingkir, Salatiga with standar provision of SNI 3144-2015.For the detection of coliform contamination, method used Most probable Number (MPN series of three tubes with Presumtive Test and Comfirmative Test, as well as the detection of Salmonella sp. with SSA (Salmonella Shigella Agar specific media. The research result that average sample of manufacturers in Sidorejo and Tingkir District have not met the standard limits of coliform contamination. The highest value of coliform contamination was >1100 APM/g and lowest value was 7 APM/g, while the Salmonella sp. detection in average yield positive result but found one sample showed negative result. Found one sample contaminated with Shigella sp. bacteria.High contamination of coliform and Salmonella was identified obtain from the tools and materials used in manufacturing process, lack of sanitation in manufacture environment, and so the individual as the manufacturer.

Use of zebrafish to study Shigella infection

Science.gov (United States)

Duggan, Gina M.

2018-01-01

ABSTRACT Shigella is a leading cause of dysentery worldwide, responsible for up to 165 million cases of shigellosis each year. Shigella is also recognised as an exceptional model pathogen to study key issues in cell biology and innate immunity. Several infection models have been useful to explore Shigella biology; however, we still lack information regarding the events taking place during the Shigella infection process in vivo. Here, we discuss a selection of mechanistic insights recently gained from studying Shigella infection of zebrafish (Danio rerio), with a focus on cytoskeleton rearrangements and cellular immunity. We also discuss how infection of zebrafish can be used to investigate new concepts underlying infection control, including emergency granulopoiesis and the use of predatory bacteria to combat antimicrobial resistance. Collectively, these insights illustrate how Shigella infection of zebrafish can provide fundamental advances in our understanding of bacterial pathogenesis and vertebrate host defence. This information should also provide vital clues for the discovery of new therapeutic strategies against infectious disease in humans. PMID:29590642

Use of zebrafish to study Shigella infection

Directory of Open Access Journals (Sweden)

Gina M. Duggan

2018-02-01

Full Text Available Shigella is a leading cause of dysentery worldwide, responsible for up to 165 million cases of shigellosis each year. Shigella is also recognised as an exceptional model pathogen to study key issues in cell biology and innate immunity. Several infection models have been useful to explore Shigella biology; however, we still lack information regarding the events taking place during the Shigella infection process in vivo. Here, we discuss a selection of mechanistic insights recently gained from studying Shigella infection of zebrafish (Danio rerio, with a focus on cytoskeleton rearrangements and cellular immunity. We also discuss how infection of zebrafish can be used to investigate new concepts underlying infection control, including emergency granulopoiesis and the use of predatory bacteria to combat antimicrobial resistance. Collectively, these insights illustrate how Shigella infection of zebrafish can provide fundamental advances in our understanding of bacterial pathogenesis and vertebrate host defence. This information should also provide vital clues for the discovery of new therapeutic strategies against infectious disease in humans.

Thermal tolerance of acid-adapted and unadapted Salmonella, Escherichia coli O157:H7, and Listeria monocytogenes in cantaloupe juice and watermelon juice.

Science.gov (United States)

Sharma, M; Adler, B B; Harrison, M D; Beuchat, L R

2005-01-01

A study was performed to determine D values of acid-adapted and unadapted cells of Salmonella, Escherichia coli O157:H7, and Listeria monocytogenes in cantaloupe juice and watermelon juice. Salmonella enterica serotype Poona, S. enterica serotype Saphra, two strains of E. coli O157:H7, and two strains of L. monocytogenes were grown in tryptic soy broth (TSB) and TSB supplemented with 1% glucose for 24 h at 37 degrees C. Decimal reduction times (D values) of cells suspended in unpasteurized cantaloupe juice and watermelon juice were determined. Acid-adapted cells of Salmonella and E. coli O157:H7, but not L. monocytogenes, had increased thermal tolerance compared with cells that were not acid-adapted. There was no correlation between soluble solids content of the two types of juice and thermal resistance. Growth of Salmonella and E. coli O157:H7 in cantaloupe juice, watermelon juice, or other acidic milieu, either in preharvest or postharvest environments, may result in cross protection to heat. The pasteurization conditions necessary to achieve elimination of pathogens from these juices would consequently have to be more severe if cells are habituated to acidic environments. Insights from this study provide guidance to developing pasteurization processes to eliminate Salmonella, E. coli O157:H7, and L. monocytogenes in cantaloupe juice and watermelon juice.

Use of Attenuated but Metabolically Competent Salmonella as a Probiotic To Prevent or Treat Salmonella Infection

Science.gov (United States)

Sabag-Daigle, Anice; Blunk, Henry M.; Gonzalez, Juan F.; Steidley, Brandi L.; Boyaka, Prosper N.

2016-01-01

Salmonella enterica is among the most burdensome of foodborne disease agents. There are over 2,600 serovars that cause a range of disease manifestations ranging from enterocolitis to typhoid fever. While there are two vaccines in use in humans to protect against typhoid fever, there are none that prevent enterocolitis. If vaccines preventing enterocolitis were to be developed, they would likely protect against only one or a few serovars. In this report, we tested the hypothesis that probiotic organisms could compete for the preferred nutrient sources of Salmonella and thus prevent or treat infection. To this end, we added the fra locus, which encodes a utilization pathway for the Salmonella-specific nutrient source fructose-asparagine (F-Asn), to the probiotic bacterium Escherichia coli Nissle 1917 (Nissle) to increase its ability to compete with Salmonella in mouse models. We also tested a metabolically competent, but avirulent, Salmonella enterica serovar Typhimurium mutant for its ability to compete with wild-type Salmonella. The modified Nissle strain became more virulent and less able to protect against Salmonella in some instances. On the other hand, the modified Salmonella strain was safe and effective in preventing infection with wild-type Salmonella. While we tested for efficacy only against Salmonella Typhimurium, the modified Salmonella strain may be able to compete metabolically with most, if not all, Salmonella serovars, representing a novel approach to control of this pathogen. PMID:27185789

Extreme Heat Resistance of Food Borne Pathogens Campylobacter jejuni, Escherichia coli, and Salmonella typhimurium on Chicken Breast Fillet during Cooking

OpenAIRE

de Jong, Aarieke E. I.; van Asselt, Esther D.; Zwietering, Marcel H.; Nauta, Maarten J.; de Jonge, Rob

2012-01-01

The aim of this research was to determine the decimal reduction times of bacteria present on chicken fillet in boiling water. The experiments were conducted with Campylobacter jejuni, Salmonella, and Escherichia coli. Whole chicken breast fillets were inoculated with the pathogens, stored overnight (4∘C), and subsequently cooked. The surface temperature reached 70∘C within 30 sec and 85∘C within one minute. Extremely high decimal reduction times of 1.90, 1.97, and 2.20 min were obtained fo...

Antimicrobial activity of lactic acid bacteria isolated from bekasam against staphylococcus aureus ATCC 25923, escherichia coli ATCC 25922, and salmonella sp

Science.gov (United States)

Sari, Melia; Suryanto, Dwi; Yurnaliza

2018-03-01

Bekasam is an Indonesian fermented food made of fish. As a fermented food, this food may contain some beneficial bacteria like lactic acid bacteria (LAB), which usually have antimicrobial properties such as organic acid, hydrogen peroxide, and a bacteriocin. A study on antimicrobial activity of LAB isolated from bekasam against some pathogenic bacteria has been conducted. The purpose of this study was to know the ability of crude bacteriocin produced LAB of bekasam against Staphylococcus aureus ATCC 25923, Escherichia coli ATCC 25922, and Salmonella sp. Bekasam sample was taken from South Sumatera. LAB isolation was done using de Man Rogosa and Sharpe agar. A bacterial colony with clear zone was selected and purified to get a single colony. The antagonistic assay of the LAB was conducted in Muller-Hinton agar Selected isolates with higher clearing zone were assayed for antibacterial effect of their crude bacteriocin of different culture incubation time of 6, 9, and 12 hours. The results showed that the crude extract bacteriocin of isolate MS2 of 9 hours culture incubation time inhibited more in Staphylococcus aureus ATCC 25923 with inhibition zone of 13.1 mm, whereas isolate MS9 of 9 hours culture incubation time inhibited more in Escherichia coli ATCC 25922 and Salmonella sp. with inhibition zone of 12.7 and 7.3 mm, respectively.

Campylobacter Immunity and Quantitative Excretion Rates in Thai Children

Science.gov (United States)

1993-01-01

vitro studies suggest that other pathogens, in- excluded if the diarrhea had lasted >3 days or jfthe patient had C 0 ~~~~~~~recently received...antibiotics. Age.smtsadciclancluding Shigella and Salmonella species and Escherichia coli. . . symptoms. and clinical and treatment data were recorded, and...ubiq- static cellulitic lesions progressing to bullae formation, skin uitous in the estuarine environment. It can cause a syndrome necrosis, necrotizin

The emergence and fate of horizontally acquired genes in Escherichia coli

NARCIS (Netherlands)

Passel, van M.W.J.; Marri, P.R.; Ochman, H.

2008-01-01

Bacterial species, and even strains within species, can vary greatly in their gene contents and metabolic capabilities. We examine the evolution of this diversity by assessing the distribution and ancestry of each gene in 13 sequenced isolates of Escherichia coli and Shigella. We focus on the

Reproducibility of microbial mutagenicity assays. I. Tests with Salmonella typhimurium and Escherichia coli using a standardized protocol

International Nuclear Information System (INIS)

Dunkel, V.C.; Zeiger, E.; Brusick, D.; McCoy, E.; McGregor, D.; Mortelmans, K.; Rosenkranz, H.S.; Simmon, V.F.

1984-01-01

The Salmonella/microsome test developed by Ames and his coworkers has been widely used in the evaluation of chemicals for genotoxic potential. Although the value of this assay is well recognized, there have been no comprehensive studies on the interlaboratory reproducibility of the method using a standardized protocol. A program was therefore initiated to compare the results obtained in four laboratories from testing a series of coded mutagens and nonmutagens using a standardized protocol. Additional objectives of this study were to compare male Fisher 344 rat, B6C3F1 mouse, and Syrian hamster liver S-9 preparations for the activation of chemicals; to compare Aroclor 1254-induced liver S-9 from all three species with the corresponding non-induced liver S-9's; and to compare the response of Escherichia coli WP-2 uvrA with the Salmonella typhimurium tester strains recommended by Ames. Since a primary use of in vitro microbial mutagenesis tests is the identification of potential carcinogens by their mutagenicity, the authors decided to compare the animal species and strains used by the National Cancer Institute/National Toxicology Program (NCI/NTP) for animal carcinogenicity studies

Microbial biosafety of pilot-scale bioreactor treating MTBE and TBA-contaminated drinking water supply.

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Schmidt, Radomir; Klemme, David A; Scow, Kate; Hristova, Krassimira

2012-03-30

A pilot-scale sand-based fluidized bed bioreactor (FBBR) was utilized to treat both methyl tert-butyl ether (MTBE) and tert-butyl alcohol (TBA) from a contaminated aquifer. To evaluate the potential for re-use of the treated water, we tested for a panel of water quality indicator microorganisms and potential waterborne pathogens including total coliforms, Escherichia coli, Salmonella and Shigella spp., Campylobacter jejuni, Aeromonas hydrophila, Legionella pneumophila, Vibrio cholerae, Yersinia enterocolytica and Mycobacterium avium in both influent and treated waters from the bioreactor. Total bacteria decreased during FBBR treatment. E. coli, Salmonella and Shigella spp., C. jejuni, V. cholerae, Y. enterocolytica and M. avium were not detected in aquifer water or bioreactor treated water samples. For those pathogens detected, including total coliforms, L. pneumophila and A. hydrophila, numbers were usually lower in treated water than influent samples, suggesting removal during treatment. The detection of particular bacterial species reflected their presence or absence in the influent waters. Copyright © 2012 Elsevier B.V. All rights reserved.

Antimicrobial-resistant Shigella infections from Iran

DEFF Research Database (Denmark)

Tajbakhsh, Mercedeh; García Migura, Lourdes; Rahbar, Mohammad

2012-01-01

Objectives: In this study, we wanted to assess the level of antimicrobial resistance, the presence of genes encoding resistance to cephalosporins and plasmid-mediated quinolone resistance (PMQR), and genetic relatedness among Shigella isolates obtained from Iranian patients. ; Methods: A total...... of 44 Shigella isolates were collected from Iranian patients admitted to Milad Hospital, Tehran, Iran, during 2008–10. Of these, 37 were serotyped and characterized by MIC determination. A subset of eight suspected extended-spectrum β-lactamase (ESBL) producers (six Shigella sonnei phase II and two...... Shigella flexneri type 1b) were examined for the presence of genes encoding cephalosporin resistance. The presence of PMQR was assessed in one S. flexneri isolate exhibiting low-level resistance to ciprofloxacin and susceptibility to nalidixic acid. PFGE was performed on 25 S. sonnei phase II isolates...

Use of ionizing radiation in elaborated products of chicken (nuggets) for control of Escherichia coli O157:H7 and salmonella enteriditis

International Nuclear Information System (INIS)

Kaempffer Ramirez, Daniela Andrea

2005-01-01

The application of ionizing energy in Chicken's Nugget of a trade mark, was studied by the University Santo Tomas in conjunction with the Chilean Nuclear Energy Commission in order to eliminate pathogens such as Escherichia coli and Salmonella enteritidis, in addition to 9 nuggets without sterilize select for RAM. Furthermore, to search the organoleptic characteristic and to Almost Chemist Analysis by means of determined doses. A total of 144 frozen Nuggets (-18 o C) were analyzed and divided into two equal groups for artificially contamination. Each Nuggets - weighting 25 g - was put into sealed polyetilene bags. The two sample groups were sterilized with an irradiation doses of 25 kGy, then inoculated with Escherichia coli O157:H7 strain ATCC 25922 ISP (Instituto de Salud Publica) and Salmonella enteritidis strain ATCC 1833-99 ISP. The samples of each group were treated with gamma irradiation (Co 60 ), with doses of 0, 0.3,0.6 and 0.9 kGy analyzing the bacteria counts after 24 hours, 30 nd 60 days post radiation. D 10 values mid-point were found with a 0,242 kGy for the Escherichia coli and 0,295 kGy for the Salmonella enteritidis. The storage time did not time exert any influence on the D 10 values. The storage time don't showed meaningful differences between the days 1, 30 and 60, in the results obtained in the mid-point D 10 value. The sensory analysis of Nugget was done by means of a trained sensorial panel who evaluated samples with 0; 0.75 and 1,5 kGy. Each person tasted the complete sample (25 g.) and evaluated; appearance, color, odor, , bitterness, texture, flavor and acceptability. Statistically, none attribute showed significant differences (p≤0.05) between the radiated samples and the control group, concluding that the level of the Nugget acceptability was qualified as very good. The Almost Chemist Analysis of Nugget was done to two groups: A control Group (without to irradiate) and a irradiated group with 1.5 kGy. It's evaluated Proteins, Fats

Enteric bacterial pathogens in children with diarrhea in Niger: diversity and antimicrobial resistance.

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Céline Langendorf

Full Text Available Although rotavirus is the leading cause of severe diarrhea among children in sub-Saharan Africa, better knowledge of circulating enteric pathogenic bacteria and their antimicrobial resistance is crucial for prevention and treatment strategies.As a part of rotavirus gastroenteritis surveillance in Maradi, Niger, we performed stool culture on a sub-population of children under 5 with moderate-to-severe diarrhea between April 2010 and March 2012. Campylobacter, Shigella and Salmonella were sought with conventional culture and biochemical methods. Shigella and Salmonella were serotyped by slide agglutination. Enteropathogenic Escherichia coli (EPEC were screened by slide agglutination with EPEC O-typing antisera and confirmed by detection of virulence genes. Antimicrobial susceptibility was determined by disk diffusion. We enrolled 4020 children, including 230 with bloody diarrhea. At least one pathogenic bacterium was found in 28.0% of children with watery diarrhea and 42.2% with bloody diarrhea. Mixed infections were found in 10.3% of children. EPEC, Salmonella and Campylobacter spp. were similarly frequent in children with watery diarrhea (11.1%, 9.2% and 11.4% respectively and Shigella spp. were the most frequent among children with bloody diarrhea (22.1%. The most frequent Shigella serogroup was S. flexneri (69/122, 56.5%. The most frequent Salmonella serotypes were Typhimurimum (71/355, 20.0%, Enteritidis (56/355, 15.8% and Corvallis (46/355, 13.0%. The majority of putative EPEC isolates was confirmed to be EPEC (90/111, 81.1%. More than half of all Enterobacteriaceae were resistant to amoxicillin and co-trimoxazole. Around 13% (46/360 Salmonella exhibited an extended-spectrum beta-lactamase phenotype.This study provides updated information on enteric bacteria diversity and antibiotic resistance in the Sahel region, where such data are scarce. Whether they are or not the causative agent of diarrhea, bacterial infections and their antibiotic

Antibacterial Activities and Possible Modes of Action of Acacia nilotica (L. Del. against Multidrug-Resistant Escherichia coli and Salmonella

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Muhammad Bilal Sadiq

2017-01-01

Full Text Available Medicinal plants are frequently used for the treatment of various infectious diseases. The objective of this study was to evaluate the antibacterial activity and mode of action of Acacia nilotica and the antibiogram patterns of foodborne and clinical strains of Escherichia coli and Salmonella. The mechanism of action of acacia extracts against E. coli and Salmonella was elucidated by observing morphological damages including cell integrity and cell membrane permeability, as well as changes in cell structures and growth patterns in kill-time experiments. The clinical isolates of E. coli and Salmonella were found resistant to more of the tested antibiotics, compared to food isolates. Minimum inhibitory concentration and minimum bactericidal concentration of acacia leaf extracts were in the ranges of 1.56–3.12 mg/mL and 3.12–6.25 mg/mL, respectively, whereas pods and bark extracts showed somewhat higher values of 3.12–6.25 mg/mL and 6.25–12.5 mg/mL, respectively, against all tested pathogens. The release of electrolytes and essential cellular constituents (proteins and nucleic acids indicated that acacia extracts damaged the cellular membrane of the pathogens. These changes corresponded to simultaneous reduction in the growth of viable bacteria. This study indicates that A. nilotica can be a potential source of new antimicrobials, effective against antibiotic-resistant strains of pathogens.

Prevalence of Salmonella serovars, Listeria monocytogenes, and Escherichia coli O157:H7 in Mediterranean ready-to-eat meat products in Jordan.

Science.gov (United States)

Osaili, Tareq M; Al-Nabulsi, Anas A; Shaker, Reyad R; Jaradat, Ziad W; Taha, Mohammad; Al-Kherasha, Mohammed; Meherat, Mervet; Holley, Richard

2014-01-01

The presence of Salmonella, Listeria monocytogenes, and Escherichia coli O157:H7 in ready-to-eat (RTE) meat products is considered a major concern for food control authorities worldwide. The aims of this study were to determine (i) the prevalence of Salmonella, L. monocytogenes, and E. coli O157:H7 in Mediterranean RTE chicken and beef (CB) products sold in Jordanian restaurants and (ii) the susceptibility of the isolates to antibiotics. A total of 1,028 samples of various types of RTE CB products (550 RTE chicken and 478 RTE beef products) were analyzed by methods described by the International Organization for Standardization followed by molecular confirmation of the isolates. The VITEK2 automated system was used for testing antibiotic susceptibility of the isolates. The overall prevalence of Salmonella serovars in RTE CB products was 0.5%, with 0.8 and 0.2% in RTE chicken and RTE beef, respectively. The overall prevalence of L. monocytogenes in RTE CB products was 2%, with 2.7 and 1.5% in RTE chicken and RTE beef products, respectively. E. coli O157:H7 was not isolated from any of the tested samples. Multidrug-resistant Salmonella and L. monocytogenes isolates were found. The majority of Salmonella isolates were sensitive to most of the tested antibiotics, and all of the isolates were resistant to more than one antibiotic. Similarly, more than 85% of L. monocytogenes isolates were sensitive to nine antibiotics, and the majority of L. monocytogenes isolates were resistant to fosfomycin and oxacillin.

Antibacterial activity of red algae (Gracilaria verrucosa) extract against Escherichia coli and Salmonella typhimurium

Science.gov (United States)

Dayuti, S.

2018-04-01

Red alga was widely used in several fields, including food, feed, phamacy and industrial point of view. The chemical analysis showed that red alga contained terpenoid, acetogenic, and aromatic compounds, which have a wide range of biological activities, such as anti-micobial, anti-inflammatory and anti-viral. The objectives of this research was to evaluate the effect of extraction solvent and time on antibacterial activity of red alga (Gracilaria verrucosa), and to explore the bioactive compound contained within Gracilaria verrucosa. The method in this study used descriptive reseach. These findings revealed that the highest inhibition activity among all extracts was obtained with the ratio of methanol:aquades (75:25) and extraction time around 72 hours against Escherichia coli and Salmonella typhimurium. The bioactive compounds of Gracilaria verrucosa tested by phytochemical analysisi consisted of flavonoid, alkaloid, and saponin. Those secondary metabolites may be approximated as antibactial substances.

Evaluation of an x-ray microprobe technique as a possible aid to detect salmonellae

International Nuclear Information System (INIS)

Richter, E.R.; Banwart, G.J.

1982-01-01

Specific bacterial antigen (Salmonella) increased in phosphorus and sqlfur after reaction with the flukrescain isothiocyanate-tagged anti-Salmonella antibody, while nonspecific antigen (Escherichia coli) did not. X-ray microprobe analysis may be useful in detecting salmonellae or other bacteria by determining increases in the elemental constituents of bacterial cells when reacted with elemental-tagged antibodies

Effects of antecedent fermentative and respiratory growth on the detection of chloramine-stressed Escherichia coil and Salmonella typhimurium.

Science.gov (United States)

Thunberg, R L; Sexstone, A J; Calabrese, J P; Bissonnette, G K

2001-08-01

In vitro laboratory studies were performed to assess the effects of antecedent growth conditions on the recovery of Escherichia coli ATCC 25922 and Salmonella typhimurium ATCC 14028 following chloramine disinfection. Six- and 18-h cultures of each organism were grown under aerobic, fermentative, and nitrate-reducing conditions prior to disinfection. At predetermined time intervals during a 10-min exposure to chloramine, survivors were surface plated on nonselective recovery media to determine C(n)t values. It was observed that nitrate-reducing growth predisposed the test organisms towards an increased sensitivity to chloramine stress over cells grown under fermentation or aerobic conditions (p < 0.01).

Biological characteristics and probiotic effect of Leuconostoc lactis strain isolated from the intestine of black porgy fish

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Wei Zhang

2013-09-01

Full Text Available A strain of lactic acid bacteria, Leuconostoc lactis, was isolated from the intestinal tract of black porgy, Sparus macrocephalus, and identified by conventional biochemical characteristics and 16S rDNA gene sequence analysis. The isolated strain had the ability of bile tolerance and resistance to low pH, and survived well in the trypsinase and pepsin solution. But the highly concentrated dose of trypsinase and pepsin affect the viability of the isolated strain. The isolate was resistant to several antibiotics, including Cephalothin, Ceftriaxone, Imipenem and Tobramycin. The isolate could autoaggregate itself and coaggregate with other bacteria in vitro. The autoaggregation percentage increased to 23.29% after 20 h of incubation. The percentage of coaggregation were respectively 31.21%, 29.44%, 10.74%, 16.49%, 24.36%, 24.41% and 20.99% for Vibrio parahaemolyticus, Listeria monocytogenes, Escherichia coli O157, Salmonella typhimurium, Shigella, Staphylococcus aureus and Proteusbacillus vulgaris after 20 h incubation of a mixed suspension. The supernatant of the strain inhibited the growth of several pathogens, such as V.parahaemolyticus, Vibrio harveyi, Vibrio alginolyticus, Staphylococcus aureus, Escherichia coli O157, Salmonella typhimurium, Bacillus subtilis, Proteusbacillus vulgaris and Shigella. These results indicated that the isolate, Leuconostoc lactis, might be an attractive candidate for perspectival strain for probiotics in marine aquaculture.

Prevalence and antimicrobial resistance among Escherichia coli and Salmonella in Ontario smallholder chicken flocks.

Science.gov (United States)

Lebert, L; Martz, S-L; Janecko, N; Deckert, A E; Agunos, A; Reid, A; Rubin, J E; Reid-Smith, R J; McEwen, S A

2018-02-01

Surveillance is an important component of an overall strategy to address antimicrobial resistant bacteria in food animals and the food chain. The poultry market has many points of entry into the Canadian food chain, and some production practices are underrepresented in terms of surveillance. For example, pathogen carriage and antimicrobial resistance surveillance data are limited in smallholder chicken flocks raised for slaughter at provincially inspected abattoirs. In Canada, antimicrobial resistance in Escherichia coli and Salmonella isolated from commercial broiler chicken flocks, slaughtered at federally inspected abattoirs, is monitored by the Canadian Integrated Program for Antimicrobial Resistance Surveillance (CIPARS). The objective of this study was to establish baseline information of antimicrobial resistance presence in E. coli and Salmonella isolated from smallholder flocks in Ontario, utilizing CIPARS collection and isolation methodologies, and to compare findings with CIPARS federally inspected abattoir data from Ontario, Canada. Five chickens per flock were sampled from 205 smallholder flocks. Of 1,025 samples, the E. coli prevalence was 99% (1,022/1,025), and 47% (483/1,022) of positive E. coli isolates were resistant to one or more of the 14 antimicrobials. Furthermore, as compared to results reported for the CIPARS commercial flocks, E. coli isolates from smallholder flocks had significantly lower resistance prevalence to six of 14 individual antimicrobials. Recovery of E. coli did not differ between federally inspected and provincially inspected flocks. Salmonella prevalence at the bird level in smallholder flocks was 0.3% (3/1,025), significantly lower (p ≪ 0.0001, 95% CI 0.080%-0.86%) than federally inspected commercial flocks. The overall differences found between the commercial and smallholder flocks may be explained by differences in poultry husbandry practices and hatchery sources. © 2017 Her Majesty the Queen in Right of Canada

Comparison of UV-C and Pulsed UV Light Treatments for Reduction of Salmonella, Listeria monocytogenes, and Enterohemorrhagic Escherichia coli on Eggs.

Science.gov (United States)

Holck, Askild L; Liland, Kristian H; Drømtorp, Signe M; Carlehög, Mats; McLEOD, Anette

2018-01-01

Ten percent of all strong-evidence foodborne outbreaks in the European Union are caused by Salmonella related to eggs and egg products. UV light may be used to decontaminate egg surfaces and reduce the risk of human salmonellosis infections. The efficiency of continuous UV-C (254 nm) and pulsed UV light for reducing the viability of Salmonella Enteritidis, Listeria monocytogenes, and enterohemorrhagic Escherichia coli on eggs was thoroughly compared. Bacterial cells were exposed to UV-C light at fluences from 0.05 to 3.0 J/cm 2 (10 mW/cm 2 , for 5 to 300 s) and pulsed UV light at fluences from 1.25 to 18.0 J/cm 2 , resulting in reductions ranging from 1.6 to 3.8 log, depending on conditions used. Using UV-C light, it was possible to achieve higher reductions at lower fluences compared with pulsed UV light. When Salmonella was stacked on a small area or shielded in feces, the pulsed UV light seemed to have a higher penetration capacity and gave higher bacterial reductions. Microscopy imaging and attempts to contaminate the interior of the eggs with Salmonella through the eggshell demonstrated that the integrity of the eggshell was maintained after UV light treatments. Only minor sensory changes were reported by panelists when the highest UV doses were used. UV-C and pulsed UV light treatments appear to be useful decontamination technologies that can be implemented in continuous processing.

Distinct Mutations Led to Inactivation of Type 1 Fimbriae Expression in Shigella spp.

Science.gov (United States)

Bravo, Verónica; Puhar, Andrea; Sansonetti, Philippe; Parsot, Claude; Toro, Cecilia S.

2015-01-01

Shigella spp. are responsible for bacillary dysentery in humans. The acquisition or the modification of the virulence plasmid encoding factors promoting entry of bacteria into and dissemination within epithelial cells was a critical step in the evolution of these bacteria from their Escherichia coli ancestor(s). Incorporation of genomic islands (GI) and gene inactivation also shaped interactions between these pathogens and their human host. Sequence analysis of the GI inserted next to the leuX tRNA gene in S. boydii, S. dysenteriae, S. flexneri, S. sonnei and enteroinvasive E. coli (EIEC) suggests that this region initially carried the fec, yjhATS and fim gene clusters. The fim cluster encoding type I fimbriae is systematically inactivated in both reference strains and clinical isolates and distinct mutations are responsible for this inactivation in at least three phylogenetic groups. To investigate consequences of the presence of fimbriae on the outcome of the interaction of Shigella with host cells, we used a S. flexneri strain harboring a plasmid encoding the E. coli fim operon. Production of fimbriae by this recombinant strain increased the ability of bacteria to adhere to and enter into epithelial cells and had no effect on their ability to disseminate from cell to cell. The observations that production of type I fimbriae increases invasion of epithelial cells and that independent mutations abolish fimbriae production in Shigella suggest that these mutations correspond to pathoadaptive events. PMID:25811616

Distinct mutations led to inactivation of type 1 fimbriae expression in Shigella spp.

Directory of Open Access Journals (Sweden)

Verónica Bravo

Full Text Available Shigella spp. are responsible for bacillary dysentery in humans. The acquisition or the modification of the virulence plasmid encoding factors promoting entry of bacteria into and dissemination within epithelial cells was a critical step in the evolution of these bacteria from their Escherichia coli ancestor(s. Incorporation of genomic islands (GI and gene inactivation also shaped interactions between these pathogens and their human host. Sequence analysis of the GI inserted next to the leuX tRNA gene in S. boydii, S. dysenteriae, S. flexneri, S. sonnei and enteroinvasive E. coli (EIEC suggests that this region initially carried the fec, yjhATS and fim gene clusters. The fim cluster encoding type I fimbriae is systematically inactivated in both reference strains and clinical isolates and distinct mutations are responsible for this inactivation in at least three phylogenetic groups. To investigate consequences of the presence of fimbriae on the outcome of the interaction of Shigella with host cells, we used a S. flexneri strain harboring a plasmid encoding the E. coli fim operon. Production of fimbriae by this recombinant strain increased the ability of bacteria to adhere to and enter into epithelial cells and had no effect on their ability to disseminate from cell to cell. The observations that production of type I fimbriae increases invasion of epithelial cells and that independent mutations abolish fimbriae production in Shigella suggest that these mutations correspond to pathoadaptive events.

Effects of cattle feeding regimen and soil management type on the fate of Escherichia coli O157:H7 and salmonella enterica serovar typhimurium in manure, manure-amended soil, and lettuce

NARCIS (Netherlands)

Franz, Eelco; van Diepeningen, Anne D; de Vos, Oscar J; van Bruggen, Ariena H C

2005-01-01

Survival of the green fluorescent protein-transformed human pathogens Escherichia coli O157:H7 and Salmonella enterica serovar Typhimurium was studied in a laboratory-simulated lettuce production chain. Dairy cows were fed three different roughage types: high-digestible grass silage plus maize

Effects of cattle feeding regimen and soil management type on the fate of Escherichia coli O157:H7 and Salmonella enterica serovar Typhimurium in manure, manure-amended soil, and lettuce

NARCIS (Netherlands)

Franz, E.; Diepeningen, van A.D.; Vos, de O.J.; Bruggen, van A.H.C.

2005-01-01

Survival of the green fluorescent protein-transformed human pathogens Escherichia coli O157:H7 and Salmonella enterica serovar Typhimurium was studied in a laboratory-simulated lettuce production chain. Dairy cows were fed three different roughage types: high-digestible grass silage plus maize

Characterization of probiotic Escherichia coli isolates with a novel pan-genome microarray

DEFF Research Database (Denmark)

Willenbrock, Hanni; Hallin, Peter Fischer; Wassenaar, Trudy

2007-01-01

of the same species are rapidly becoming available, allowing for the definition and characterization of a whole species as a population of genomes - the 'pan-genome'. Results: Using 32 Escherichia coli and Shigella genome sequences we estimate the pan- and core genome of the species. We designed a high...

Status of vaccine research and development for Shigella.

Science.gov (United States)

Mani, Sachin; Wierzba, Thomas; Walker, Richard I

2016-06-03

Shigella are gram-negative bacteria that cause severe diarrhea and dysentery. In 2013, Shigella infections caused an estimated 34,400 deaths in children less than five years old and, in 2010, an estimated 40,000 deaths in persons older than five years globally. New disease burden estimates from newly deployed molecular diagnostic assays with increased sensitivity suggest that Shigella-associated morbidity may be much greater than previous disease estimates from culture-based methods. Primary prevention of this disease should be based on universal provision of potable water and sanitation methods and improved personal and food hygiene. However, an efficacious and low-cost vaccine would complement and accelerate disease reduction while waiting for universal access to water, sanitation, and hygiene improvements. This review article provides a landscape of Shigella vaccine development efforts. No vaccine is yet available, but human and animal challenge-rechallenge trials with virulent Shigella as well as observational studies in Shigella-endemic areas have shown that the incidence of disease decreases following Shigella infection, pointing to biological feasibility of a vaccine. Immunity to Shigella appears to be strain-specific, so a vaccine that covers the most commonly detected strains (i.e., S. flexneri 2a, 3a, 6, and S. sonnei) or a vaccine using cross-species conserved antigens would likely be most effective. Vaccine development and testing may be accelerated by use of animal models, such as the guinea pig keratoconjunctivitis or murine pneumonia models. Because there is no correlate of protection, however, human studies will be necessary to evaluate vaccine efficacy prior to deployment. A diversity of Shigella vaccine constructs are under development, including live attenuated, formalin-killed whole-cell, glycoconjugate, subunit, and novel antigen vaccines (e.g., Type III secretion system and outer membrane proteins). Copyright © 2016 World Health Organization

Serotyping, PCR, phage-typing and antibiotic sensitivity testing of Salmonella serovars isolated from urban drinking water supply systems of Nepal

DEFF Research Database (Denmark)

Bhatta, D.R.; Bangtrakulnonth, A.; Tishyadhigama, P.

2007-01-01

Aims: To study the occurrence and diversity of Salmonella serovars in urban water supply systems of Nepal. Methods and Results: Occurrence of Salmonella was detected in 42 out of 300 water samples by enrichment culture technique in selenite F broth followed by plating on Salmonella Shigella agar...... isolates of Salm. Enteritidis indicated the presence of one of the ESBL genes, blaSHV, whereas the genes blaTEM and blaCTX were absent. Conclusions: The microbiological quality of the urban water supply is poor and indicates possibility of fatal outbreaks of enteric fever and related infections in Nepal....... Significance and Impact of the Study: The present study will be useful in water borne disease control and prevention strategy formulation in Nepal and in the global context....

Antigen-Specific lgA B Memory Cell Responses to Shigella Antigens Elicited in Volunteers Immunized with Live Attenuated Shigella flexneri 2a Oral Vaccine Candidates

Science.gov (United States)

2011-01-01

167. [10] E.V. Oaks, T.L. Hale, S.B. Formal, Serum immune response to Shigella protein antigens in rhesus monkeys and humans infected with Shigella ...cell responses to Shigella antigens elicited in volunteers immunized with live attenuated Shigella flexneri 2a oral vaccine candidates J.K. Simona,b... Shigella ;. B cell memory; Immunoglobulin lgA; Mucosal immunity Abstract We studied the induction of antigen-specific lgA memory B cells (BM) in

EFSA BIOHAZ Panel (EFSA Panel on Biological Hazards), 2014. Scientific Opinion on the risk posed by pathogens in food of non-animal origin. Part 2 (Salmonella Yersinia, Shigella and Norovirus in bulb and stem vegetables, and carrots)

DEFF Research Database (Denmark)

Hald, Tine; Baggesen, Dorte Lau

, Shigella and Norovirus were considered in the context of the whole food chain. Available estimates of their occurrence in these vegetables were evaluated together with mitigation options relating to prevention of contamination and the relevance of microbiological criteria. Emphasis is given to vegetable...... of food safety management systems including Good Agricultural Practices (GAP), Good Hygiene Practices (GHP) and Good Manufacturing Practices (GMP) should be the primary objectives of producers of bulb and stem vegetables as well as carrots. Considering the limited evidence for both the occurrence......Bulb and stem vegetables as well as carrots may be minimally processed to obtain ready-to-eat products, and these steps include selection, washing, cleaning, cutting, packaging and storage. Risk factors for the contamination of bulb and stem vegetables as well as carrots with Salmonella, Yersinia...

Effect of wild-type Shigella species and attenuated Shigella vaccine candidates on small intestinal barrier function, antigen trafficking, and cytokine release.

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Maria Fiorentino

Full Text Available Bacterial dysentery due to Shigella species is a major cause of morbidity and mortality worldwide. The pathogenesis of Shigella is based on the bacteria's ability to invade and replicate within the colonic epithelium, resulting in severe intestinal inflammatory response and epithelial destruction. Although the mechanisms of pathogenesis of Shigella in the colon have been extensively studied, little is known on the effect of wild-type Shigella on the small intestine and the role of the host response in the development of the disease. Moreover, to the best of our knowledge no studies have described the effects of apically administered Shigella flexneri 2a and S. dysenteriae 1 vaccine strains on human small intestinal enterocytes. The aim of this study was to assess the coordinated functional and immunological human epithelial responses evoked by strains of Shigella and candidate vaccines on small intestinal enterocytes. To model the interactions of Shigella with the intestinal mucosa, we apically exposed monolayers of human intestinal Caco2 cells to increasing bacterial inocula. We monitored changes in paracellular permeability, examined the organization of tight-junctions and the pro-inflammatory response of epithelial cells. Shigella infection of Caco2 monolayers caused severe mucosal damage, apparent as a drastic increase in paracellular permeability and disruption of tight junctions at the cell-cell boundary. Secretion of pro-inflammatory IL-8 was independent of epithelial barrier dysfunction. Shigella vaccine strains elicited a pro-inflammatory response without affecting the intestinal barrier integrity. Our data show that wild-type Shigella infection causes a severe alteration of the barrier function of a small intestinal cell monolayer (a proxy for mucosa and might contribute (along with enterotoxins to the induction of watery diarrhea. Diarrhea may be a mechanism by which the host attempts to eliminate harmful bacteria and transport them

Search for Enterohaemorrhagic Escherichia coli O157:H7 and ...

African Journals Online (AJOL)

Enterohaemorrhagic Escherichia coli (EHEC) O157:H7 and Salmonella enterica are important zoonotic bacteria responsible for enteric infections in humans. The present study investigated the possible role of kittens in the zoonotic transmission of antimicrobial resistant EHEC O157 and Salmonella enterica to human using ...

A Brief History of Shigella.

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Lampel, Keith A; Formal, Samuel B; Maurelli, Anthony T

2018-01-01

The history of Shigella , the causative agent of bacillary dysentery, is a long and fascinating one. This brief historical account starts with descriptions of the disease and its impact on human health from ancient time to the present. Our story of the bacterium starts just before the identification of the dysentery bacillus by Kiyoshi Shiga in 1898 and follows the scientific discoveries and principal scientists who contributed to the elucidation of Shigella pathogenesis in the first 100 years. Over the past century, Shigella has proved to be an outstanding model of an invasive bacterial pathogen and has served as a paradigm for the study of other bacterial pathogens. In addition to invasion of epithelial cells, some of those shared virulence traits include toxin production, multiple-antibiotic resistance, virulence genes encoded on plasmids and bacteriophages, global regulation of virulence genes, pathogenicity islands, intracellular motility, remodeling of host cytoskeleton, inflammation/polymorphonuclear leukocyte signaling, apoptosis induction/inhibition, and "black holes" and antivirulence genes. While there is still much to learn from studying Shigella pathogenesis, what we have learned so far has also contributed greatly to our broader understanding of bacterial pathogenesis.

An In Vitro Chicken Gut Model Demonstrates Transfer of a Multidrug Resistance Plasmid from Salmonella to Commensal Escherichia coli.

Science.gov (United States)

Card, Roderick M; Cawthraw, Shaun A; Nunez-Garcia, Javier; Ellis, Richard J; Kay, Gemma; Pallen, Mark J; Woodward, Martin J; Anjum, Muna F

2017-07-18

The chicken gastrointestinal tract is richly populated by commensal bacteria that fulfill various beneficial roles for the host, including helping to resist colonization by pathogens. It can also facilitate the conjugative transfer of multidrug resistance (MDR) plasmids between commensal and pathogenic bacteria which is a significant public and animal health concern as it may affect our ability to treat bacterial infections. We used an in vitro chemostat system to approximate the chicken cecal microbiota, simulate colonization by an MDR Salmonella pathogen, and examine the dynamics of transfer of its MDR plasmid harboring several genes, including the extended-spectrum beta-lactamase bla CTX-M1 We also evaluated the impact of cefotaxime administration on plasmid transfer and microbial diversity. Bacterial community profiles obtained by culture-independent methods showed that Salmonella inoculation resulted in no significant changes to bacterial community alpha diversity and beta diversity, whereas administration of cefotaxime caused significant alterations to both measures of diversity, which largely recovered. MDR plasmid transfer from Salmonella to commensal Escherichia coli was demonstrated by PCR and whole-genome sequencing of isolates purified from agar plates containing cefotaxime. Transfer occurred to seven E. coli sequence types at high rates, even in the absence of cefotaxime, with resistant strains isolated within 3 days. Our chemostat system provides a good representation of bacterial interactions, including antibiotic resistance transfer in vivo It can be used as an ethical and relatively inexpensive approach to model dissemination of antibiotic resistance within the gut of any animal or human and refine interventions that mitigate its spread before employing in vivo studies. IMPORTANCE The spread of antimicrobial resistance presents a grave threat to public health and animal health and is affecting our ability to respond to bacterial infections

Aptasensors for rapid detection of Escherichia coli O157:H7 and Salmonella typhimurium

Science.gov (United States)

Wu, Wen-he; Li, Min; Wang, Yue; Ouyang, Hou-xian; Wang, Lin; Li, Ci-xiu; Cao, Yu-chen; Meng, Qing-he; Lu, Jian-xin

2012-11-01

Herein we reported the development of aptamer-based biosensors (aptasensors) based on label-free aptamers and gold nanoparticles (AuNPs) for detection of Escherichia coli ( E. coli) O157:H7 and Salmonella typhimurium. Target bacteria binding aptamers are adsorbed on the surface of unmodified AuNPs to capture target bacteria, and the detection was accomplished by target bacteria-induced aggregation of the aptasensor which is associated as red-to-purple color change upon high-salt conditions. By employing anti- E. coli O157:H7 aptamer and anti- S. typhimurium aptamer, we developed a convenient and rapid approach that could selectively detect bacteria without specialized instrumentation and pretreatment steps such as cell lysis. The aptasensor could detect as low as 105colony-forming units (CFU)/ml target bacteria within 20 min or less and its specificity was 100%. This novel method has a great potential application in rapid detection of bacteria in the near future.

MOLECULAR-PHYLOGENETIC CHARACTERIZATION AND ANTIMICROBIAL RESISTANCE OF Escherichia coli ISOLATED FROM GOATS WITH DIARRHEA

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Rodrigo Almeida Guimarães

2015-10-01

Full Text Available Neonatal diarrhea determines significant changes in feed conversion, causing productivity loss in caprine herds. The antimicrobial resistance in bacteria is characterized as an important public health issue; therefore, Escherichia coli may be characterized as an important pathogen due to expressing virulence mechanisms responsible for significant clinical conditions in humans and animals. The present study evaluated the presence of E. coli among 117 caprine fecal samples and analyzed the isolates for antimicrobial resistance. Suggestive colonies were submitted to biochemical screening followed by genotypic group determination and phylogenetic analysis; further, the samples were submitted to antimicrobials susceptibility test. E. coli, Salmonella spp, Shigella sonnei and Enterobacter aerogenes were identified. E. coli isolates were phylogenetically classified as B2 (9/39, D (19/39, B1 (7/39 e A (4/29 groups. The analysis of the isolates also revealed the presence of K99 (04/39 and Stx (02/39 virulence factors. Antimicrobial susceptibility test revealed sensitive isolates to Chloramphenicol, Streptomycin, Amoxicillin and Ciprofloxacin, being all resistant to Lincomycin, Vancomycin and Penicillin. The results support the need of establishing restricted protocols for antimicrobial use, a fundamental procedure for health improvement in Brazilian caprine herds.

Genotypic and Phenotypic Characterization of Enterotoxigenic Escherichia coli Strains Isolated from Peruvian Children

Science.gov (United States)

2010-09-01

adhesion, ETEC elabo- rates one or both of two enterotoxins: heat-labile toxin (LT), a protein multimer which shares many features with cholera toxin...pathogens (Shigella, Salmonella. Vibrio , Campylo/Jacter, Gi- ardia Iamblia, Cryptosporidium, and rotavirus) by conventional methods. Five lactose...studies performed in developing coun- tries: 38% in children ɚ years of age in Nicaragua (19), 33% in children ə year of age in Mexico (6), and 18% in

Fate of Escherichia coli O157:H7 and Salmonella in soil and lettuce roots as affected by potential home gardening practices.

Science.gov (United States)

Erickson, Marilyn C; Liao, Jean; Payton, Alison S; Webb, Cathy C; Ma, Li; Zhang, Guodong; Flitcroft, Ian; Doyle, Michael P; Beuchat, Larry R

2013-12-01

The survival and distribution of enteric pathogens in soil and lettuce systems were investigated in response to several practices (soil amendment supplementation and reduced watering) that could be applied by home gardeners. Leaf lettuce was grown in manure compost/top soil (0:5, 1:5 or 2:5 w/w) mixtures. Escherichia coli O157:H7 or Salmonella was applied at a low or high dose (10(3) or 10(6) colony-forming units (CFU) mL(-1) ) to the soil of seedlings and mid-age plants. Supplementation of top soil with compost did not affect pathogen survival in the soil or on root surfaces, suggesting that nutrients were not a limiting factor. Salmonella populations on root surfaces were 0.7-0.8 log CFU g(-1) lower for mid-age plants compared with seedlings. E. coli O157:H7 populations on root surfaces were 0.8 log CFU g(-1) lower for mid-age plants receiving 40 mL of water compared with plants receiving 75 mL of water on alternate days. Preharvest internalization of E. coli O157:H7 and Salmonella into lettuce roots was not observed at any time. Based on the environmental conditions and high pathogen populations in soil used in this study, internalization of Salmonella or E. coli O157:H7 into lettuce roots did not occur under practices that could be encountered by inexperienced home gardeners. © 2013 Society of Chemical Industry.

Molecular characterization of multidrug-resistant Shigella spp. of food origin.

Science.gov (United States)

Ahmed, Ashraf M; Shimamoto, Tadashi

2015-02-02

Shigella spp. are the causative agents of food-borne shigellosis, an acute enteric infection. The emergence of multidrug-resistant clinical isolates of Shigella presents an increasing challenge for clinicians in the treatment of shigellosis. Several studies worldwide have characterized the molecular basis of antibiotic resistance in clinical Shigella isolates of human origin, however, to date, no such characterization has been reported for Shigella spp. of food origin. In this study, we characterized the genetic basis of multidrug resistance in Shigella spp. isolated from 1600 food samples (800 meat products and 800 dairy products) collected from different street venders, butchers, retail markets, and slaughterhouses in Egypt. Twenty-four out of 27 Shigella isolates (88.9%) showed multidrug resistance phenotypes to at least three classes of antimicrobials. The multidrug-resistant Shigella spp. were as follows: Shigella flexneri (66.7%), Shigella sonnei (18.5%), and Shigella dysenteriae (3.7%). The highest resistance was to streptomycin (100.0%), then to kanamycin (95.8%), nalidixic acid (95.8%), tetracycline (95.8%), spectinomycin (93.6%), ampicillin (87.5%), and sulfamethoxazole/trimethoprim (87.5%). PCR and DNA sequencing were used to screen and characterize integrons and antibiotic resistance genes. Our results indicated that 11.1% and 74.1% of isolates were positive for class 1 and class 2 integrons, respectively. Beta-lactamase-encoding genes were identified in 77.8% of isolates, and plasmid-mediated quinolone resistance genes were identified in 44.4% of isolates. These data provide useful information to better understand the molecular basis of antimicrobial resistance in Shigella spp. To the best of our knowledge, this is the first report of the molecular characterization of antibiotic resistance in Shigella spp. isolated from food. Copyright © 2014 Elsevier B.V. All rights reserved.

Infección por Campylobacter y Shigella como causa de Diarrea Aguda Infecciosa en niños menores de dos años en el Distrito de la Victoria, Lima-Perú

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María Perales D

2002-10-01

Full Text Available Objetivo: Determinar la frecuencia de Campylobacter y Shigella como agentes etiológicos en diarrea aguda acuosa en niños menores de dos años atendidos en 4 centros de salud del distrito de La Victoria (Lima, Perú Materiales y métodos: en este estudio transversal analítico se realizaron coprocultivos bajo técnica microbiológica estándar a los niños menores de dos años con diarrea aguda acuosa atendidos en 4 centros de salud de La Victoria entre abril y octubre de 2001. Fueron excluidos aquellos niños con disentería y aquellos que habían recibido tratamiento antibiótico previo a la atención. Resultados: se estudiaron un total de 248 casos, de los cuales 48 (19,4% coprocultivos fueron positivos: 33 (13.3% a Campylobacter, 12 (4,8% a Shigella y 3 (1,2% a Salmonella. No se identificaron otros patógenos. Las especies de Shigella identificadas fueron: Shigella flexneri en 9 (75,0% casos y Shigella sonnei en 3 (25,0% La única especie de Campylobacter identificada fue Campylobacter jejuni siendo el biotipo I el más frecuente (84,6% Conclusiones: Campylobacter y Shigella juegan un importante papel como agentes etiológicos causantes de diarrea aguda acuosa en niños menores de dos años.

Assessing the growth of Escherichia coli O157:H7 and Salmonella in spinach, lettuce, parsley and chard extracts at different storage temperatures.

Science.gov (United States)

Posada-Izquierdo, G; Del Rosal, S; Valero, A; Zurera, G; Sant'Ana, A S; Alvarenga, V O; Pérez-Rodríguez, F

2016-06-01

The objective of this work was to study the growth potential of Escherichia coli O157:H7 and Salmonella spp. in leafy vegetable extracts at different temperature conditions. Cocktails of five strains of E. coli O157:H7 and of Salmonella enterica were used. Inoculated aqueous vegetable extracts were incubated at 8, 10, 16 and 20°C during 21 days. Microbial growth was monitored using Bioscreen C(®) . In spinach extract, results showed that for E. coli O157:H7 and Salmonella significant differences (P parsley showed the lowest values of μabs , below 0·008 h(-1) . The coefficients of variance (CoV) calculated for the different replicates evidenced that at low temperature (8°C) a more variable behaviour of both pathogens is expected (CoV > 180%). This study provides evidence that aqueous extracts from vegetable tissues can result in distinct growth niche producing different response in various types of vegetables. Finally, these results can be used as basis to establish risk rankings of pathogens and leafy vegetable matrices with relation to their potential growth. © 2016 The Society for Applied Microbiology.

Combining Shigella Tn-seq data with gold-standard E. coli gene deletion data suggests rare transitions between essential and non-essential gene functionality.

Science.gov (United States)

Freed, Nikki E; Bumann, Dirk; Silander, Olin K

2016-09-06

Gene essentiality - whether or not a gene is necessary for cell growth - is a fundamental component of gene function. It is not well established how quickly gene essentiality can change, as few studies have compared empirical measures of essentiality between closely related organisms. Here we present the results of a Tn-seq experiment designed to detect essential protein coding genes in the bacterial pathogen Shigella flexneri 2a 2457T on a genome-wide scale. Superficial analysis of this data suggested that 481 protein-coding genes in this Shigella strain are critical for robust cellular growth on rich media. Comparison of this set of genes with a gold-standard data set of essential genes in the closely related Escherichia coli K12 BW25113 revealed that an excessive number of genes appeared essential in Shigella but non-essential in E. coli. Importantly, and in converse to this comparison, we found no genes that were essential in E. coli and non-essential in Shigella, implying that many genes were artefactually inferred as essential in Shigella. Controlling for such artefacts resulted in a much smaller set of discrepant genes. Among these, we identified three sets of functionally related genes, two of which have previously been implicated as critical for Shigella growth, but which are dispensable for E. coli growth. The data presented here highlight the small number of protein coding genes for which we have strong evidence that their essentiality status differs between the closely related bacterial taxa E. coli and Shigella. A set of genes involved in acetate utilization provides a canonical example. These results leave open the possibility of developing strain-specific antibiotic treatments targeting such differentially essential genes, but suggest that such opportunities may be rare in closely related bacteria.

A new piece of the Shigella Pathogenicity puzzle: spermidine accumulation by silencing of the speG gene [corrected].

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Marialuisa Barbagallo

Full Text Available The genome of Shigella, a gram negative bacterium which is the causative agent of bacillary dysentery, shares strong homologies with that of its commensal ancestor, Escherichia coli. The acquisition, by lateral gene transfer, of a large plasmid carrying virulence determinants has been a crucial event in the evolution towards the pathogenic lifestyle and has been paralleled by the occurrence of mutations affecting genes, which negatively interfere with the expression of virulence factors. In this context, we have analysed to what extent the presence of the plasmid-encoded virF gene, the major activator of the Shigella regulon for invasive phenotype, has modified the transcriptional profile of E. coli. Combining results from transcriptome assays and comparative genome analyses we show that in E. coli VirF, besides being able to up-regulate several chromosomal genes, which potentially influence bacterial fitness within the host, also activates genes which have been lost by Shigella. We have focused our attention on the speG gene, which encodes spermidine acetyltransferase, an enzyme catalysing the conversion of spermidine into the physiologically inert acetylspermidine, since recent evidence stresses the involvement of polyamines in microbial pathogenesis. Through identification of diverse mutations, which prevent expression of a functional SpeG protein, we show that the speG gene has been silenced by convergent evolution and that its inactivation causes the marked increase of intracellular spermidine in all Shigella spp. This enhances the survival of Shigella under oxidative stress and allows it to better face the adverse conditions it encounters inside macrophage. This is supported by the outcome of infection assays performed in mouse peritoneal macrophages and of a competitive-infection assay on J774 macrophage cell culture. Our observations fully support the pathoadaptive nature of speG inactivation in Shigella and reveal that the accumulation

Identificación de hongos fitopatógenos y presencia de salmonella sp en compost de plantas de tratamiento de aguas residuales

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Jorge Antonio Silva-Leal

2007-01-01

Full Text Available This study shows adapted methodologies to identify of phytopathogens fungi (Botrytis spp, Fusarium sp., Phytophthora sp., Rhizoctonia sp and bacteria as Salmonella sp. Presence of mentioned microorganism was evaluated in compost samples produced from dewatering primary sludge, generated in Cañaveralejo Wastewater Treatment Plant -WWTPC. The adapted methodologies shown that the appropriate dilutions for isolation of phytopathogens fungi in compost are 10-3,10-4and 10-5 and the most appropriated selective culture medium for the salmonella sp identification is the Salmonella-Shigella agar. Fusarium sp was the only phytopathogens fungi founded in compost; Salmonella sp was also founded. Therefore, it is recommended the quantification in order to define the concentrations that can cause health problems; additionally, it is necessary sanitization of compost for use in agriculture, mainly in industrial crops.

Prevalence of fecal shedding of Salmonella organisms among captive green iguanas and potential public health implications.

Science.gov (United States)

Burnham, B R; Atchley, D H; DeFusco, R P; Ferris, K E; Zicarelli, J C; Lee, J H; Angulo, F J

1998-07-01

To determine prevalence of fecal shedding of Salmonella organisms among captive green iguanas (Iguana iguana). Cohort study. 12 captive green iguanas. Iguanas were isolated in an environmental chamber, and fecal samples were collected weekly for 10 consecutive weeks. Samples were incubated aerobically in tetrathionate broth for 18 to 24 hours. Aliquots were then transferred to Hektoen and Salmonella-Shigella agar plates and incubated for an additional 18 to 24 hours. Isolated colonies were subcultured on nutrient agar slants, and Salmonella isolates were serogrouped and serotyped. All 12 iguanas were found to be shedding Salmonella organisms at least once during the study, and multiple serotypes were isolated from 7 of the 12. Salmonella organisms were isolated from 88 of 106 (83%) fecal samples; 21 samples contained multiple Salmonella serotypes. Overall, 11 Salmonella serotypes were identified. In 74 of 100 instances, when a particular Salmonella serotype was isolated from an individual iguana, the same serotype was also isolated from a subsequent fecal sample from that iguana. Results suggested that most iguanas have a stable mixture of Salmonella serotypes in their intestinal tracts and intermittently or continuously shed Salmonella organisms in their feces. Veterinarians should advise their clients on precautions for reducing the risk of acquiring these organisms from their pets. Public health officials trying to determine whether an iguana is the source of a specific Salmonella serotype that caused infection in human patients should submit at least 3 fecal samples collected from the iguana 1 week apart for bacterial culture.

Bacteriophage-based Probiotic Preparation for Managing Shigella Infections

Science.gov (United States)

2015-04-16

The preparation (designated “ShigActive”) is a bacteriophage cocktail that specifically targets Shigella spp. (significant diarrhea-causing pathogens...phages lytic for Shigella , and we have developed a murine model in which the in vivo efficacy of our 1. REPORT DATE (DD-MM-YYYY) 4. TITLE AND...10-Apr-2013 Approved for Public Release; Distribution Unlimited Final Report: Bacteriophage-based Probiotic Preparation for Managing Shigella

Detection and analysis of CRISPRs of Shigella.

Science.gov (United States)

Guo, Xiangjiao; Wang, Yingfang; Duan, Guangcai; Xue, Zerun; Wang, Linlin; Wang, Pengfei; Qiu, Shaofu; Xi, Yuanlin; Yang, Haiyan

2015-01-01

The recently discovered CRISPRs (Clustered regularly interspaced short palindromic repeats) and Cas (CRISPR-associated) proteins are a novel genetic barrier that limits horizontal gene transfer in prokaryotes and the CRISPR loci provide a historical view of the exposure of prokaryotes to a variety of foreign genetic elements. The aim of study was to investigate the occurrence and distribution of the CRISPRs in Shigella. A collection of 61 strains of Shigella were screened for the existence of CRISPRs. Three CRISPR loci were identified among 61 shigella strains. CRISPR1/cas loci are detected in 49 strains of shigella. Yet, IS elements were detected in cas gene in some strains. In the remaining 12 Shigella flexneri strains, the CRISPR1/cas locus is deleted and only a cas3' pseudo gene and a repeat sequence are present. The presence of CRISPR2 is frequently accompanied by the emergence of CRISPR1. CRISPR3 loci were present in almost all strains (52/61). The length of CRISPR arrays varied from 1 to 9 spacers. Sequence analysis of the CRISPR arrays revealed that few spacers had matches in the GenBank databases. However, one spacer in CRISPR3 loci matches the cognate cas3 genes and no cas gene was present around CRISPR3 region. Analysis of CRISPR sequences show that CRISPR have little change which makes CRISPR poor genotyping markers. The present study is the first attempt to determine and analyze CRISPRs of shigella isolated from clinical patients.

Prevalence and characterization of Listeria monocytogenes, Salmonella and Shiga toxin-producing Escherichia coli isolated from small Mexican retail markets of queso fresco.

Science.gov (United States)

Soto Beltran, Marcela; Gerba, Charles P; Porto Fett, Anna; Luchansky, John B; Chaidez, Cristobal

2015-01-01

Queso fresco (QF) is a handmade cheese consumed and produced in Latin America. In Mexico, QF production is associated with a microbiological risk. The aim of the study was to determine the incidence and characterization of Listeria monocytogenes, Salmonella spp., and Shiga toxin-producing Escherichia coli (STEC) in QF from retail markets of the north-western State of Sinaloa, Mexico, and to assess the effect of physicochemical parameters on Listeria presence. A total of 75 QF samples were obtained. L. monocytogenes, E. coli, and coliforms were detected in 9.3, 94, and 100%, respectively. Salmonella was not detected. STEC isolates showed virulence genes. Microbial loads were above the maximum values recommended by the Official Mexican Standards. Physicochemical parameters such as water activity (aw), moisture content, pH, and salinity played a role in Listeria prevalence in QF. Rigorous control in QF made in Culiacan, Mexico is needed to reduce the risk of foodborne pathogens.

Progress and pitfalls in Shigella vaccine research

Science.gov (United States)

Barry, Eileen M.; Pasetti, Marcela F.; Sztein, Marcelo B.; Fasano, Alessio; Kotloff, Karen L.; Levine, Myron M.

2013-01-01

Renewed awareness of the significant morbidity and mortality that Shigella causes among young children in developing countries combined with technological innovations in vaccinology has led to the development of novel vaccine strategies in the past five years. Along with advancement of classical vaccines in clinical trials and new sophisticated measurements of immunological responses, much new data has been produced lending promise to the potential for production of safe and effective Shigella vaccines. Herein we review the recent progress in Shigella vaccine development within the framework of persistent obstacles. PMID:23419287

MICROBIAL CONTAMINATION OF STREET VENDED FOODS FROM A UNIVERSITY CAMPUS IN BANGLADESH.

Science.gov (United States)

Islam, Sufia; Nasrin, Nishat; Rizwan, Farhana; Nahar, Lutfun; Bhowmik, Adity; Esha, Sayma Afrin; Talukder, Kaisar Ali; Akter, Mahmuda; Roy, Ajoy; Ahmed, Muniruddin

2015-05-01

The microbiological quality of street vended food samples from Dhaka, Bangladesh was evaluated. The objective of the study was to identify the presence of common pathogens (Escherichia coli, Shigella spp, Salmonella and Vibrio spp) and to describe the molecular characterization of E coli, a commonly found pathogen in various street foods. Fifty food samples were collected from fixed and mobile vendors from two sampling locations (Mohakhali and Aftabnagar) in Dhaka city, Bangladesh. The tested samples included deep fried and fried snacks; quick lunch items; pickles; fruit chutney; baked items; spicy, sour and hot snacks etc: Juices, tamarind water and plain drinking water were also tested. Sterile polythene bags were used for collecting 200 g of each category of samples. They were tested for the presence of microorganisms following conventional microbiological processes. Biochemical tests followed by serology were done for the confirmation of Shigella and Salmonella. Serological reaction was carried out for confirmation of Vibrio spp. DNA was isolated for the molecular characterization to detect the pathogenic E. coli by polymerase chain reaction (PCR). Out of 50 food samples, six (12%) were confirmed to contain different species of E. coli and Shigella. Molecular characterization of E. coli revealed that three samples were contaminated with enteroaggregative E. coli (EAEC) and one was contaminated with enterotoxigenic E. coli (ETEC). Shigellaflexneri X variant was detected in one food item and Shigella flexneri 2a was found in drinking water. All these enteric pathogens could be the potential cause for foodborne illnesses.

Growth of Escherichia coli O157:H7, Non-O157 Shiga Toxin-Producing Escherichia coli , and Salmonella in Water and Hydroponic Fertilizer Solutions.

Science.gov (United States)

Shaw, Angela; Helterbran, Kara; Evans, Michael R; Currey, Christopher

2016-12-01

The desire for local, fresh produce year round is driving the growth of hydroponic growing systems in the United States. Many food crops, such as leafy greens and culinary herbs, grown within hydroponics systems have their root systems submerged in recirculating nutrient-dense fertilizer solutions from planting through harvest. If a foodborne pathogen were introduced into this water system, the risk of contamination to the entire crop would be high. Hence, this study was designed to determine whether Escherichia coli O157:H7, non-O157 Shiga toxin-producing E. coli , and Salmonella were able to survive and reproduce in two common hydroponic fertilizer solutions and in water or whether the bacteria would be killed or suppressed by the fertilizer solutions. All the pathogens grew by 1 to 6 log CFU/ml over a 24-h period, depending on the solution. E. coli O157:H7 reached higher levels in the fertilizer solution with plants (3.12 log CFU/ml), whereas non-O157 Shiga toxin-producing E. coli and Salmonella reached higher levels in the fertilizer solution without plants (1.36 to 3.77 log CFU/ml). The foodborne pathogens evaluated here survived for 24 h in the fertilizer solution, and populations grew more rapidly in these solutions than in plain water. Therefore, human pathogens entering the fertilizer solution tanks in hydroponic systems would be expected to rapidly propagate and spread throughout the system and potentially contaminate the entire crop.

The genome and proteome of a virulent Escherichia coli O157:H7 bacteriophage closely resembling Salmonella phage Felix O1

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Waddell Thomas E

2009-04-01

Full Text Available Abstract Based upon whole genome and proteome analysis, Escherichia coli O157:H7-specific bacteriophage (phage wV8 belongs to the new myoviral genus, "the Felix O1-like viruses" along with Salmonella phage Felix O1 and Erwinia amylovora phage φEa21-4. The genome characteristics of phage wV8 (size 88.49 kb, mol%G+C 38.9, 138 ORFs, 23 tRNAs are very similar to those of phage Felix O1 (86.16 kb, 39.0 mol%G+C, 131 ORFs and 22 tRNAs and, indeed most of the proteins have their closest homologs within Felix O1. Approximately one-half of the Escherichia coli O157:H7 mutants resistant to phage wV8 still serotype as O157:H7 indicating that this phage may recognize, like coliphage T4, two different surface receptors: lipopolysaccharide and, perhaps, an outer membrane protein.

Occurrence and antimicrobial resistance of pathogenic Escherichia coli and Salmonella spp. in retail raw table eggs sold for human consumption in Enugu state, Nigeria

Science.gov (United States)

Okorie-Kanu, O. Josephine; Ezenduka, E. Vivienne; Okorie-Kanu, C. Onwuchokwe; Ugwu, L. Chinweokwu; Nnamani, U. John

2016-01-01

Aim: This study was conducted to investigate the occurrence of pathogenic Escherichia coli and Salmonella species in retail raw table eggs sold for human consumption in Enugu State and to determine the resistance of these pathogens to antimicrobials commonly used in human and veterinary practices in Nigeria. Materials and Methods: A total of 340 raw table eggs comprising 68 composite samples (5 eggs per composite sample) were collected from five selected farms (13 composite samples from the farms) and 10 retail outlets (55 composite samples from the retail outlets) in the study area over a period of 4-month (March-June, 2014). The eggs were screened for pathogenic E. coli and Salmonella species following standard procedures within 24 h of sample collection. Isolates obtained were subjected to in-vitro antimicrobial susceptibility test with 15 commonly used antimicrobials using the disk diffusion method. Results: About 37 (54.4%) and 7 (10.3%) of the 68 composite samples were positive for pathogenic E. coli and Salmonella species, respectively. The shells showed significantly higher (p0.05). The organisms obtained showed a multiple drug resistance. They were completely resistant to nitrofurantoin, sulfamethoxazole/trimethoprim, penicillin G and oxacillin. In addition to these, Salmonella spp. also showed 100% resistance to tetracycline. The pathogenic E. coli isolates obtained were 100% susceptible to gentamicin, neomycin, ciprofloxacin, and amoxicillin-clavulanic acid while Salmonella spp. showed 100% susceptibility to erythromycin, neomycin, and rifampicin. Both organisms showed varying degrees of resistance to streptomycin, amoxicillin, vancomycin, and doxycycline. Conclusion: From the results of the study, it can be concluded that the raw table eggs marketed for human consumption in Enugu State, Nigeria is contaminated with pathogenic E. coli and Salmonella species that showed multiple drug resistance to antimicrobial agents commonly used in veterinary and human

Fate of Escherichia coli O157:H7 and Salmonella on whole strawberries and blueberries of two maturities under different storage conditions.

Science.gov (United States)

Nguyen, Thao P; Friedrich, Loretta M; Danyluk, Michelle D

2014-07-01

Strawberries and blueberries harvested at or near full-ripe maturity tend to be less firm and more susceptible to bruising during harvest and transport. The objective of this research was to determine the fate of Escherichia coli O157:H7 and Salmonella on bruised and intact surfaces of whole strawberries and blueberries at shipping (2°C) and retail display (15.5°C) temperatures. Strawberries and blueberries were either purchased from a supermarket or were harvested immediately prior to use; they were bruised using established protocols, were spot inoculated, and were incubated at 2 and 15.5°C. Strawberries, subjected to modified atmospheres, were further transferred to bags and were sealed in with an initial atmosphere of ca. 10% CO2 and 5% O2. Strawberries were sampled at 0, 2, 5, and 24 h and on days 3 and 7; blueberries were sampled on days 0, 1, 3, and 7. After stomaching, samples were enumerated on nonselective and selective media, and populations were recorded as log CFU per berry. At both storage temperatures, population declines for both E. coli O157:H7 and Salmonella were seen under all conditions for strawberries. At 2 ± 2°C, E. coli O157:H7 and Salmonella populations on blueberries declined over 7 days under all conditions. At 15.5 ± 2°C, E. coli O157:H7 populations declined; however, Salmonella populations initially declined but increased to populations near or above initial populations over 7 days on blueberries. No overall significant differences were observed between bruised and intact treatments or between the two maturity levels for strawberries and blueberries. Modified atmospheric conditions did not affect the behavior of E. coli O157:H7 and Salmonella on strawberries at both temperatures. This research indicates that E. coli O157:H7 and Salmonella do not grow on strawberries at shipping or retail display temperatures, even when they are harvested at a maturity prone to bruising; however, Salmonella growth may occur on bruised full ripe

Development of a Live Attenuated Bivalent Oral Vaccine Against Shigella sonnei Shigellosis and Typhoid Fever.

Science.gov (United States)

Wu, Yun; Chakravarty, Sumana; Li, Minglin; Wai, Tint T; Hoffman, Stephen L; Sim, B Kim Lee

2017-01-15

Shigella sonnei and Salmonella Typhi cause significant morbidity and mortality. We exploited the safety record of the oral, attenuated S. Typhi vaccine (Ty21a) by using it as a vector to develop a bivalent oral vaccine to protect against S. sonnei shigellosis and typhoid fever. We recombineered the S. sonnei form I O-antigen gene cluster into the Ty21a chromosome to create Ty21a-Ss, which stably expresses S. sonnei form I O antigen. To enhance survivability in the acid environment of the stomach, we created an acid-resistant strain, Ty21a-AR-Ss, by inserting Shigella glutaminase-glutamate decarboxylase systems coexpressed with S. sonnei form I O-antigen gene. Mice immunized intranasally with Ty21a-AR-Ss produced antibodies against S. sonnei and S. Typhi, and survived lethal intranasal S. sonnei challenge. This paves the way for proposed good manufacturing practices manufacture and clinical trials intended to test the clinical effectiveness of Ty21a-AR-Ss in protecting against S. sonnei shigellosis and typhoid fever, as compared with the current Ty21a vaccine. © The Author 2016. Published by Oxford University Press for the Infectious Diseases Society of America. All rights reserved. For permissions, e-mail journals.permissions@oup.com.

21 CFR 866.3660 - Shigella spp. serological reagents.

Science.gov (United States)

2010-04-01

... (CONTINUED) MEDICAL DEVICES IMMUNOLOGY AND MICROBIOLOGY DEVICES Serological Reagents § 866.3660 Shigella spp...), used in serological tests to identify Shigella spp. from cultured isolates. The identification aids in...

Green fluorescent protein labeling of Listeria, Salmonella, and Escherichia coli O157:H7 for safety-related studies.

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Li Ma

Full Text Available Many food safety-related studies require tracking of introduced foodborne pathogens to monitor their fate in complex environments. The green fluorescent protein (GFP gene (gfp provides an easily detectable phenotype so has been used to label many microorganisms for ecological studies. The objectives of this study were to label major foodborne pathogens and related bacteria, including Listeria monocytogenes, Listeria innocua, Salmonella, and Escherichia coli O157:H7 strains, with GFP and characterize the labeled strains for stability of the GFP plasmid and the plasmid's effect on bacterial growth. GFP plasmids were introduced into these strains by a CaCl(2 procedure, conjugation or electroporation. Stability of the label was determined through sequential propagation of labeled strains in the absence of selective pressure, and rates of plasmid-loss were calculated. Stability of the GFP plasmid varied among the labeled species and strains, with the most stable GFP label observed in E. coli O157:H7. When grown in nonselective media for two consecutive subcultures (ca. 20 generations, the rates of plasmid loss among labeled E. coli O157:H7, Salmonella and Listeria strains ranged from 0%-30%, 15.8%-99.9% and 8.1%-93.4%, respectively. Complete loss (>99.99% of the plasmid occurred in some labeled strains after five consecutive subcultures in the absence of selective pressure, whereas it remained stable in others. The GFP plasmid had an insignificant effect on growth of most labeled strains. E. coli O157:H7, Salmonella and Listeria strains can be effectively labeled with the GFP plasmid which can be stable in some isolates for many generations without adversely affecting growth rates.

Physical Covering for Control of Escherichia coli O157:H7 and Salmonella spp. in Static and Windrow Composting Processes

Science.gov (United States)

Yossa, Irene; Macarisin, Dumitru; Millner, Patricia

2015-01-01

This study investigated the effect of a 30-cm covering of finished compost (FC) on survival of Escherichia coli O157:H7 and Salmonella spp. in active static and windrow composting systems. Feedstocks inoculated with E. coli O157:H7 (7.41 log CFU/g) and Salmonella (6.46 log CFU/g) were placed in biosentry tubes (7.5-cm diameter, 30-cm height) at three locations: (i and ii) two opposing sides at the interface between the FC cover layer (where present) and the feedstock material (each positioned approximately 10 cm below the pile's surface) and (iii) an internal location (top) (approximately 30 cm below the surface). On specific sampling days, surviving populations of inoculated E. coli O157:H7 and Salmonella, generic E. coli, and coliforms in compost samples were determined. Salmonella spp. were reduced significantly within 24 h in windrow piles and were below the detection limit after 3 and 7 days at internal locations of windrow and static piles containing FC covering, respectively. Likewise, E. coli O157:H7 was undetectable after 1 day in windrow piles covered with finished compost. Use of FC as a covering layer significantly increased the number of days that temperatures in the windrows remained ≥55°C at all locations and in static piles at internal locations. These time-temperature exposures resulted in rapid reduction of inoculated pathogens, and the rate of bacterial reduction was rapid in windrow piles. The sample location significantly influenced the survival of these pathogens at internal locations compared to that at interface locations of piles. Finished compost covering of compost piles aids in the reduction of pathogens during the composting process. PMID:25576620

Shigella Vulvovaginitis in Childhood: A Case Report

Science.gov (United States)

Rajkumar, Solomon; Narayanaswamy, Geetha; Laude, Teresita Awa

1979-01-01

Vulvovaginitis due to Shigella sonnei is reported in a four-year-old child. Shigella sonnei as a rare primary pathogen causing vulvovaginitis, the prolonged nature of the clinical course, and the therapeutic implications of increasing resistance to ampicillin by the organism are discussed. PMID:395325

Identification and Characterization of T5-Like Bacteriophages Representing Two Novel Subgroups from Food Products

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Domonkos Sváb

2018-02-01

Full Text Available During recent years, interest in the use of bacteriophages as biocontrol agents against foodborne pathogens has increased, particularly for members of the family Enterobacteriaceae, with pathogenic Escherichia coli, Shigella, and Salmonella strains among them. Here, we report the isolation and characterisation of 12 novel T5-like bacteriophages from confiscated food samples. All bacterophages effectively lysed E. coli K-12 strains and were able to infect pathogenic E. coli strains representing enterohaemorrhagic (EHEC, enteropathogenic (EPEC, enterotoxigenic (ETEC, and enteroinvasive (EIEC pathotypes, Shigella dysenteriae, S. sonnei strains, as well as multidrug-resistant (MDR E. coli and multiple strains representing different Salmonella enterica serovars. All the bacteriophages exhibited Siphoviridae morphology. Whole genome sequencing of the novel T5-like bacteriophages showed that they represent two distinct groups, with the genome-based grouping correlating to the different host spectra. As these bacteriophages are of food origin, their stability and lack of any virulence genes, as well as their broad and mutually complementary host spectrum makes these new T5-like bacteriophages valuable candidates for use as biocontrol agents against foodborne pathogenic enterobacteria.

Effectiveness of superheated steam for inactivation of Escherichia coli O157:H7, Salmonella Typhimurium, Salmonella Enteritidis phage type 30, and Listeria monocytogenes on almonds and pistachios.

Science.gov (United States)

Ban, Ga-Hee; Kang, Dong-Hyun

2016-03-02

This study was undertaken to evaluate the effectiveness of superheated steam (SHS) on the inactivation of Escherichia coli O157:H7, Salmonella Typhimurium, Salmonella Enteritidis phage type (PT) 30 and Listeria monocytogenes on almonds and in-shell pistachios and to determine the effect of superheated steam heating on quality by measuring color and texture changes. Almonds and in-shell pistachios inoculated with four foodborne pathogens were treated with saturated steam (SS) at 100 °C and SHS at 125, 150, 175, and 200 °C for various times. Exposure of almonds and pistachios to SHS for 15 or 30s at 200 °C achieved >5l og reductions among all tested pathogens without causing significant changes in color values or texture parameters (P>0.05). For both almonds and pistachios, acid and peroxide values (PV) following SS and SHS treatment for up to 15s and 30s, respectively, were within the acceptable range (PV<1.0 meq/kg). These results show that thermal application of 200 °C SHS treatment for 15s and 30s did not affect the quality of almonds and pistachios, respectively. Therefore, SHS treatment is a very promising alternative technology for the tree nuts industry by improving inactivation of foodborne pathogens on almonds and pistachios while simultaneously reducing processing time. Copyright © 2016 Elsevier B.V. All rights reserved.

Antibiotic resistant Salmonella and Escherichia coli isolated from ...

African Journals Online (AJOL)

Objective: To characterise and investigate antimicrobial resistance of Esherichia coli and salmonella strains isolated from indigenous Gallus gallus in a leading slaughterhouse/market outlet in Nairobi-Kenya. Design: A repeated cross sectional study and based on random sampling was used. Setting: The study was carried ...

ANTIBACTERIAL EFFECT OF GARLIC (ALLIUM SATIVUM AND GINGER (ZINGIBER OFFICINALE AGAINST STAPHYLOCOCCUS AUREUS, SALMONELLA TYPHI, ESCHERICHIA COLI AND BACILLUS CEREUS

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Bandna Chand

2013-02-01

Full Text Available Antibacterial activity of extracts of Allium sativum (garlic and Zingiber officinale (ginger has been evaluated against four different bacteria namely Escherichia coli, Salmonella Typhi, Staphylococcus aureus and Bacillus cereus. Two methods were used to determine the antimicrobial activity of garlic and ginger extracts namely disk diffusion method and agar well diffusion method. Garlic extract exhibited excellent antibacterial activity against all four test organisms while ginger extract showed antibacterial activity against Bacillus cereus and Staphylococcus aureus only. In addition, agar well diffusion method showed higher zone in inhibition when compared with the zone of inhibition produced by the spice of same concentration against the test microorganism by disk diffusion method. Antibiotic sensitivity of the four different bacteria was tested with commercially available antibiotics namely Ciprofloxacin; Oxytetracycline; Vancomycin; Streptomycin; Gentamicin; Tetracycline; Novobiocin; Amikacin and Penicillin G. Penicillin G produced the highest zone of inhibition of 40.00±0.00against Staphylococcus aureus and the lowest zone of inhibition of 0.00±0.00against Escherichia coli.

Multidrug resistant shigella flexneri infection simulating intestinal intussusception

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Srirangaraj Sreenivasan

2016-01-01

Full Text Available Shigella enteritis remains an important cause of mortality and morbidity in all age groups, in developing as well as developed countries. Owing to the emerging resistance to multiple antibiotics among Shigella spp., it has been recognized as a major global public health concern and warrants constant monitoring of its resistance pattern. We report a case of segmental ileitis caused by non.-ESBL producing multidrug resistant Shigella flexneri in an infant clinically mimicking intussusception, which was effectively treated by ceftriaxone.

Sepsis por shigella flexneri

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César Cabrera C

2005-04-01

Full Text Available Se presenta un caso raro de sepsis por Shigella flexneri en una paciente de 45 años de edad quien estando hospitalizada para el estudio de un tumor cerebral, requirió el uso de manitol y dosis altas de corticoides; luego de ello presenta deposiciones líquidas con moco y sangre, desarrolla síndrome de respuesta inflamatoria sistémica, luego se aísla Shigella flexneri en el hemocultivo; recibió tratamiento antibiótico con ciprofloxacina. Se describen las características del caso y se comenta de acuerdo con la revisión de literatura.

In vitro selection of RNA aptamer specific to Salmonella typhimurium.

Science.gov (United States)

Han, Seung Ryul; Lee, Seong-Wook

2013-06-28

Salmonella is a major foodborne pathogen that causes a variety of human diseases. Development of ligands directly and specifically binding to the Salmonella will be crucial for the rapid detection of, and thus for efficient protection from, the virulent bacteria. In this study, we identified a RNA aptamer-based ligand that can specifically recognize Salmonella Typhimurium through SELEX technology. To this end, we isolated and characterized an RNase-resistant RNA aptamer that bound to the OmpC protein of Salmonella Typhimurium with high specificity and affinity (Kd ~ 20 nM). Of note, the selected aptamer was found to specifically bind to Salmonella Typhimurium, but neither to Gram-positive bacteria (Staphylococcus aureus) nor to other Gram-negative bacteria (Escherichia coli O157:H7). This was evinced by aptamer-immobilized ELISA and aptamer-linked precipitation experiments. This Salmonella species-specific aptamer could be useful as a diagnostic ligand against pathogen-caused foodborne sickness.

Study of the action of {sup 60}Co gamma radiation on Salmonella poona, Escherichia coli and Alicyclobacillus acidoterrestris in mango pulp; Estudo da acao da radiacao gama de {sup 60}Co sobre Salmonella poona, Escherichia coli e Alicyclobacillus acidoterrestris em polpa de manga congelada

Energy Technology Data Exchange (ETDEWEB)

Pereira, Marco Antonio dos Santos

2009-07-01

The application of non-thermal treatments has proven effective in inhibiting bacteria such as Salmonella and Escherichia coli. Mango is a fruit of national consumption with a great exportation potential. Meanwhile, outbreaks of food borne disease related to mango consumption caused mistrust on the degree of food security offered by the product. The objective of this work was to establish the radioresistance of bacteria Escherichia coli, Salmonella Poona and Alicyclobacillus acidoterrestris on the mango pulp by the calculation of the D10 values and to know the radiation effect on the sensory characteristics of the fruit pulp. The microbiological profile of frozen mango pulp available at the local market was also established using conventional methods of plating and Most Probable Number (MPN). The pulps experimentally inoculated with the bacteria listed above were irradiated with doses of 0, 1, 2, 3, 4 and 5kGy in a {sup 60}Co source. The sensory analysis was performed using a dose of 5 kGy, using the triangular test and the test of acceptance with hedonic scale. The results of this study show that the quality of mango pulp sell in the local market is not satisfactory in accordance with the standards established by the Brazilian law and the literature, showing the need of using other tools to achieve acceptable levels of quality. The D10 values obtained are in the range of 1.01 and 1.09kGy for E. coli ATCC 8739, 0.60 and 0.98kGy for S. poona and 0.72 e 0.88kGy for A. acidoterrestris respectively. The triangular test showed that a 5kGy radiation dose changed the sensory characteristics of mango pulp. Nevertheless, sensory analysis of a food product prepared with the irradiated pulp obtained good acceptance in the attributes of global appearance, flavor and aroma. (author)

Study of the action of {sup 60}Co gamma radiation on Salmonella poona, Escherichia coli and Alicyclobacillus acidoterrestris in mango pulp;Estudo da acao da radiacao gama de {sup 60}Co sobre Salmonella poona, Escherichia coli and Alicyclobacillus acidoterrestris em polpa de manga congelada

Energy Technology Data Exchange (ETDEWEB)

Pereira, Marco Antonio dos Santos

2009-07-01

The application of non-thermal treatments has proven effective in inhibiting bacteria such as Salmonella and Escherichia coli. Mango is a fruit of national consumption with a great exportation potential. Meanwhile, outbreaks of foodborne disease related to mango consumption caused mistrust on the degree of food security offered by the product. The objective of this work was to establish the radioresistance of bacteria Escherichia coli, Salmonella Poona and Alicyclobacillus acidoterrestris on the mango pulp by the calculation of the D10 values and to know the radiation effect on the sensory characteristics of the fruit pulp. The microbiological profile of frozen mango pulp available at the local market was also established using conventional methods of plating and Most Probable Number (MPN). The pulps experimentally inoculated with the bacteria listed above were irradiated with doses of 0, 1, 2, 3, 4 and 5 kGy in a {sup 60}Co source. The sensory analysis was performed using a dose of 5 kGy, using the triangular test and the test of acceptance with hedonic scale. The results of this study show that the quality of mango pulp sell in the local market is not satisfactory in accordance with the standards established by the Brazilian law and the literature, showing the need of using other tools to achieve acceptable levels of quality. The D10 values obtained are in the range of 1.01 and 1.09 kGy for E. coli ATCC 8739, 0.60 and 0.98 kGy for S. poona and 0.72 and 0.88 kGy for A. acidoterrestris respectively. The triangular test showed that a 5 kGy radiation dose changed the sensory characteristics of mango pulp. Nevertheless, sensory analysis of a food product prepared with the irradiated pulp obtained good acceptance in the attributes of global appearance, flavor and aroma. (author)

New vaccine strategies against enterotoxigenic Escherichia coli: II: Enhanced systemic and secreted antibody responses against the CFA/I fimbriae by priming with DNA and boosting with a live recombinant Salmonella vaccine

Directory of Open Access Journals (Sweden)

M.O. Lásaro

1999-02-01

Full Text Available The induction of systemic (IgG and mucosal (IgA antibody responses against the colonization factor I antigen (CFA/I of enterotoxigenic Escherichia coli (ETEC was evaluated in mice primed with an intramuscularly delivered CFA/I-encoding DNA vaccine followed by two oral immunizations with a live recombinant Salmonella typhimurium vaccine strain expressing the ETEC antigen. The booster effect induced by the oral immunization was detected two weeks and one year after the administration of the DNA vaccine. The DNA-primed/Salmonella-boosted vaccination regime showed a synergistic effect on the induced CFA/I-specific systemic and secreted antibody levels which could not be attained by either immunization strategy alone. These results suggest that the combined use of DNA vaccines and recombinant Salmonella vaccine strains can be a useful immunization strategy against enteric pathogens.

Riboregulators: Fine-Tuning Virulence in Shigella.

Science.gov (United States)

Fris, Megan E; Murphy, Erin R

2016-01-01

Within the past several years, RNA-mediated regulation (ribo-regulation) has become increasingly recognized for its importance in controlling critical bacterial processes. Regulatory RNA molecules, or riboregulators, are perpetually responsive to changes within the micro-environment of a bacterium. Notably, several characterized riboregulators control virulence in pathogenic bacteria, as is the case for each riboregulator characterized to date in Shigella. The timing of virulence gene expression and the ability of the pathogen to adapt to rapidly changing environmental conditions is critical to the establishment and progression of infection by Shigella species; ribo-regulators mediate each of these important processes. This mini review will present the current state of knowledge regarding RNA-mediated regulation in Shigella by detailing the characterization and function of each identified riboregulator in these pathogens.

PERSEPSI MASYARAKAT TERHADAP PENYAKIT SHIGELLA (DISENTRI DI JAKARTA UTARA

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Siti Sapardiyah Santoso

2012-09-01

Full Text Available Penelitian telah dilakukan di Kecamatan Tanjung Priok dan Kecamatan Koja, Jakarta Utara, mengenai perilaku dan sosial budaya dalam penanggulangan penyakit shigella (disentri. Tulisan ini merupakan bagian dari penelitian tersebut. Tujuan penelitian untuk mengetahui persepsi masyarakat Jakarta Utara mengenai penyakit shigella (disentri. Rancangan penelitian cross sectional. Responden berumur diatas 22 tahun laki-laki dan perempuan dipilih secara acak. Pengumpulan data dilakukan dengan wawancara menggunakan koesioner terstruktur kepada 500 responden (250 laki-laki dan 250 wanita. Analisis data menggunakan SPSS. Hasil, menurut responden laki-laki dan perempuan. Masyarakat menganggap serius penyakit shigella (disentri berkisar antara 72,4% - 77,6%. Masyarakat cukup peduli terhadap penyakit tersebut berkisar antara 59,6% - 63,2%. Masyarakat menganggap sakit shigella (disentri lebih berat dari penyakit diare lainnya berkisar antara 67,2% - 71,2%. Kelompok mudah terserang penyakit tersebut adalah balita berkisar antara 85,2% - 86,6%, anak 6 -14 tahun berkisar antara 79,2% -80,8%, bayi 0-1 tahun berkisar antara 79,6% - 86,4%. Lama waktu sembuh yang mengatakan seminggu berkisar antara 54,0% - 55,6%. Penyakit tersebut berpengaruh terhadap sosial ekonomi keluarga dan dianggap serius berkisar antara 52,4% - 53,6%. Biaya bila sakit shigella (disentri ringan maupun berat dianggap mahal berkisar antara 33, 6% - 39,2%. Dari penelitian ini dapat disimpulkan bahwa masyarakat menganggap peduli dan serius terhadap penyakit shigella (disentri karena dianggap lebih parah dari diare lainnya dan pengobatannya dianggap mahal baik shigella (disentri ringan maupun berat.

Inactivation of Escherichia coli O157:H7, salmonellae, and Campylobacter jejuni in raw ground beef by gamma irradiation

International Nuclear Information System (INIS)

Clavero, M.R.S.; Monk, J.D.; Beuchat, L.R.; Doyle, M.P.; Brackett, R.E.

1994-01-01

Raw ground beef patties inoculated with stationary-phase cells of Escherichia coli O157:H7, salmonellae, or Campylobacter jejuni were subjected to gamma irradiation (60Co) treatment, with doses ranging from 0 to 2.52 kGy. The influence of two levels of fat (8 to 14% [low fat] and 27 to 28% [high fat]) and temperature (frozen [-17 to -15 degrees C] and refrigerated [3 to 5 degrees C]) on the inactivation of each pathogen by irradiation was investigated. In ascending order of irradiation resistance, the D10 values ranged from 0.175 to 0.235 kGy (C. jejuni), from 0.241 to 0.307 kGy (E. coli O157:H7), and from 0.618 to 0.800 kGy (salmonellae). Statistical analysis revealed that E. coli O157:H7 had a significantly (P 0.05) higher D10 value when irradiated at -17 to -15 degrees C than when irradiated at 3 to 5 degrees C. Regardless of the temperature during irradiation, the level of fat did not have a significant effect on the D10 value. Salmonellae behaved like E. coli O157:H7 in low-fat beef, but temperature did not have a significant effect when the pathogen was irradiated in high-fat ground beef. Significantly higher D10 values were calculated for C. jejuni irradiated in frozen than in refrigerated low-fat beef. C. jejuni was more resistant to irradiation in low-fat beef than in high-fat beef when treatment was at -17 to -15 degrees C. Regardless of the fat level and temperature during inactivation, these pathogens were highly sensitive to gamma irradiation. An applied dose of 2.5 kGy would be sufficient to kill 10(8.1) E. coli O157:H7, 10(3.1) salmonellae, and 10(10.6) C. jejuni, resulting in a high probability of complete inactivation of populations much higher than those occasionally present in ground beef patties

Effect of gamma radiation on the reduction of Salmonella strains, Listeria monocytogenes, and Shiga toxin-producing Escherichia coli and sensory evaluation of minimally processed spinach (Tetragonia expansa).

Science.gov (United States)

Rezende, Ana Carolina B; Igarashi, Maria Crystina; Destro, Maria Teresa; Franco, Bernadette D G M; Landgraf, Mariza

2014-10-01

This study evaluated the effects of irradiation on the reduction of Shiga toxin-producing Escherichia coli (STEC), Salmonella strains, and Listeria monocytogenes, as well as on the sensory characteristics of minimally processed spinach. Spinach samples were inoculated with a cocktail of three strains each of STEC, Salmonella strains, and L. monocytogenes, separately, and were exposed to gamma radiation doses of 0, 0.2, 0.4, 0.6, 0.8, and 1.0 kGy. Samples that were exposed to 0.0, 1.0, and 1.5 kGy and kept under refrigeration (4°C) for 12 days were submitted to sensory analysis. D10 -values ranged from 0.19 to 0.20 kGy for Salmonella and from 0.20 to 0.21 for L. monocytogenes; for STEC, the value was 0.17 kGy. Spinach showed good acceptability, even after exposure to 1.5 kGy. Because gamma radiation reduced the selected pathogens without causing significant changes in the quality of spinach leaves, it may be a useful method to improve safety in the fresh produce industry.

Presence of non-O157 Shiga toxin-producing Escherichia coli, enterotoxigenic E. coli, enteropathogenic E. coli and Salmonella in fresh beetroot (Beta vulgaris L.) juice from public markets in Mexico.

Science.gov (United States)

Gómez-Aldapa, Carlos A; Rangel-Vargas, Esmeralda; Bautista-De León, Haydee; Castro-Rosas, Javier

2014-10-01

Unpasteurized juice has been associated with foodborne illness outbreaks for many years. Beetroot is a vegetable grown all over the world in temperate areas. In Mexico beetroot is consumed cooked in salads or raw as fresh unpasteurized juices. No data about the microbiological quality or safety of unpasteurized beetroot juices are available. Indicator bacteria, diarrheagenic Escherichia coli pathotypes (DEP) and Salmonella frequencies were determined for fresh unpasteurized beetroot juice from restaurants. One hundred unpasteurized beetroot juice samples were collected from public markets in Pachuca, Mexico. Frequencies in these samples were 100%, 75%, 53%, 9% and 4% of positive samples, for coliform bacteria, fecal coliforms, E. coli, DEP and Salmonella, respectively. Identified DEP included enterotoxigenic E. coli (ETEC), enteropathogenic E. coli (EPEC) and non-O157 Shiga toxin-producing E. coli (STEC). Identified Salmonella serotypes included Typhimurium and Enteritidis. This is the first report of microbiological quality and atypical EPEC, ETEC, non-O157 STEC and Salmonella isolation from fresh raw beetroot juice in Mexico. Fresh raw beetroot juice from markets is very probably an important factor contributing to the endemicity of atypical EPEC, ETEC, non-O157 STEC and Salmonella-related gastroenteritis in Mexico. © 2014 Society of Chemical Industry.

Study of the action of 60Co gamma radiation on Salmonella poona, Escherichia coli and Alicyclobacillus acidoterrestris in mango pulp

International Nuclear Information System (INIS)

Pereira, Marco Antonio dos Santos

2009-01-01

The application of non-thermal treatments has proven effective in inhibiting bacteria such as Salmonella and Escherichia coli. Mango is a fruit of national consumption with a great exportation potential. Meanwhile, outbreaks of food borne disease related to mango consumption caused mistrust on the degree of food security offered by the product. The objective of this work was to establish the radioresistance of bacteria Escherichia coli, Salmonella Poona and Alicyclobacillus acidoterrestris on the mango pulp by the calculation of the D10 values and to know the radiation effect on the sensory characteristics of the fruit pulp. The microbiological profile of frozen mango pulp available at the local market was also established using conventional methods of plating and Most Probable Number (MPN). The pulps experimentally inoculated with the bacteria listed above were irradiated with doses of 0, 1, 2, 3, 4 and 5kGy in a 60 Co source. The sensory analysis was performed using a dose of 5 kGy, using the triangular test and the test of acceptance with hedonic scale. The results of this study show that the quality of mango pulp sell in the local market is not satisfactory in accordance with the standards established by the Brazilian law and the literature, showing the need of using other tools to achieve acceptable levels of quality. The D10 values obtained are in the range of 1.01 and 1.09kGy for E. coli ATCC 8739, 0.60 and 0.98kGy for S. poona and 0.72 e 0.88kGy for A. acidoterrestris respectively. The triangular test showed that a 5kGy radiation dose changed the sensory characteristics of mango pulp. Nevertheless, sensory analysis of a food product prepared with the irradiated pulp obtained good acceptance in the attributes of global appearance, flavor and aroma. (author)

Study of the action of 60Co gamma radiation on Salmonella poona, Escherichia coli and Alicyclobacillus acidoterrestris in mango pulp

International Nuclear Information System (INIS)

Pereira, Marco Antonio dos Santos

2009-01-01

The application of non-thermal treatments has proven effective in inhibiting bacteria such as Salmonella and Escherichia coli. Mango is a fruit of national consumption with a great exportation potential. Meanwhile, outbreaks of foodborne disease related to mango consumption caused mistrust on the degree of food security offered by the product. The objective of this work was to establish the radioresistance of bacteria Escherichia coli, Salmonella Poona and Alicyclobacillus acidoterrestris on the mango pulp by the calculation of the D10 values and to know the radiation effect on the sensory characteristics of the fruit pulp. The microbiological profile of frozen mango pulp available at the local market was also established using conventional methods of plating and Most Probable Number (MPN). The pulps experimentally inoculated with the bacteria listed above were irradiated with doses of 0, 1, 2, 3, 4 and 5 kGy in a 60 Co source. The sensory analysis was performed using a dose of 5 kGy, using the triangular test and the test of acceptance with hedonic scale. The results of this study show that the quality of mango pulp sell in the local market is not satisfactory in accordance with the standards established by the Brazilian law and the literature, showing the need of using other tools to achieve acceptable levels of quality. The D10 values obtained are in the range of 1.01 and 1.09 kGy for E. coli ATCC 8739, 0.60 and 0.98 kGy for S. poona and 0.72 and 0.88 kGy for A. acidoterrestris respectively. The triangular test showed that a 5 kGy radiation dose changed the sensory characteristics of mango pulp. Nevertheless, sensory analysis of a food product prepared with the irradiated pulp obtained good acceptance in the attributes of global appearance, flavor and aroma. (author)

Changing patterns of drug-resistant Shigella isolates in egypt.

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Abd-Elmeged, Ghada M; Khairy, Rasha M; Abo-Eloyoon, Sahar M; Abdelwahab, Sayed F

2015-06-01

The emergence of multidrug resistance (MDR) is a serious problem in treating shigellosis. There are limited existing data examining the change in the antimicrobial resistance profile of Shigella in Egypt. We previously reported that 58% of the Shigella isolates in Egypt were resistant to at least one member of the three different antimicrobial groups. This study was performed to determine the antimicrobial resistance profile of Shigella, determine their possible mechanisms of resistance, and compare their resistance profile to those reported 20 years ago. Stool samples were collected from 500 subjects and processed for the isolation and identification of Shigella. The susceptibility of the isolates to 11 different antimicrobials was determined using the disc diffusion method. Of 500 stool cultures, 24 (4.8%) samples were positive for Shigella. There was a high percentage of resistance to ampicillin (88%), tetracycline (83%), and sulfamethoxazole-trimethoprim (75%). Also, there was a moderate percentage of resistance to chloramphenicol (46%), streptomycin (42%), ceftazidime (33%), and cefotaxime (25%). A lower percentage of resistance was recorded for amikacin, nalidixic acid (17% each), and ofloxacin (7%), while no resistance was found to ciprofloxacin (0%). Twenty-one of the isolates (88%) were resistant to at least three different antimicrobial groups (indicating MDR). The average number of antimicrobial agents to which the Shigella isolates were resistant was 4.3±1.4, while it was 3.4±1.5 in the same locality in 1994. These data demonstrate that there is a marked increase in MDR and change in the resistance patterns of Shigella over the past 20 years.

Prevalence of pathogenic bacteria in street vended ready-to-eat meats in Windhoek, Namibia.

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Shiningeni, Daphney; Chimwamurombe, Percy; Shilangale, Renatus; Misihairabgwi, Jane

2018-05-31

To determine the prevalence of pathogenic bacteria in street vended ready-to-eat meats in Windhoek, Namibia, a total of 96 street vended ready to eat meat samples were evaluated. Prevalences of 42%, 52%, 15%, 6% and 83% were observed for Escherichia coli, Staphylococcus aureus, Listeria monocytogenes, Shigella and Enterobacteriaceae respectively, while the highest aerobic plate counts were 7.74 Log 10 cfu/g, 5.67 Log 10 cfu/g, 5.12 Log 10 cfu/g , 4.56 Log 10 cfu/g, 3.3 Log 10 cfu/g, 5.75 Log 10 cfu/g respectively. Unsatisfactory microbial levels were 32% for aerobic plate count, 26% for Enterobacteriaceae, 35% for Escherichia coli, 11% for Listeria monocytogenes, 7% for Staphylococcus aureus and 6% for Shigella. Salmonella was detected in 11% and 40% of samples from two suburbs. The unsatisfactory microbiological quality of some ready-to-eat meats necessitates the provision of training on food safety and hygiene to street vendors for consumer protection. Copyright © 2018 Elsevier Ltd. All rights reserved.

Co-administration of rIpaB domain of Shigella with rGroEL of S. Typhi enhances the immune responses and protective efficacy against Shigella infection.

Science.gov (United States)

Chitradevi, Sekar Tamil Selvi; Kaur, Gurpreet; Uppalapati, Sivaramakrishna; Yadav, Anandprakash; Singh, Dependrapratap; Bansal, Anju

2015-11-01

Shigella species cause severe bacillary dysentery in humans and are associated with high morbidity and mortality. The Invasion plasmid antigen (IpaB) protein, which is conserved across all Shigella spp., induces macrophage cell death and is required to invade host cells. The present study evaluates the immunogenicity and protective efficacy of the recombinant (r) domain region of IpaB (rIpaB) of S. flexneri. rIpaB was administered either alone or was co-administered with the rGroEL (heat shock protein 60) protein from S. Typhi as an adjuvant in a mouse model of intranasal immunization. The IpaB domain region (37 kDa) of S. flexneri was amplified from an invasion plasmid, cloned, expressed in BL21 Escherichia coli cells and purified. Immunization with the rIpaB domain alone stimulated both humoral and cell-mediated immune responses. Furthermore, robust antibody (IgG, IgA) and T-cell responses were induced when the rIpaB domain was co-administered with rGroEL. Antibody isotyping revealed higher IgG1 and IgG2a antibody titers and increased interferon-gamma (IFN-γ) secretion in the co-administered group. Immunization of mice with the rIpaB domain alone protected 60%-70% of the mice from lethal infection by S. flexneri, S. boydii and S. sonnei, whereas co-administration with rGroEL increased the protective efficacy to 80%-85%. Organ burden and histopathological studies also revealed a significant reduction in lung infection in the co-immunized mice compared with mice immunized with the rIpaB domain alone. This study emphasizes that the co-administration of the rIpaB domain and rGroEL protein improves immune responses in mice and increases protective efficacy against Shigella infection. This is also the first report to evaluate the potential of the GroEL (Hsp 60) protein of S. Typhi as an adjuvant molecule, thereby overcoming the need for commercial adjuvants.

Shift in serotype distribution of Shigella species in China, 2003-2013.

Science.gov (United States)

Qiu, S; Xu, X; Yang, C; Wang, J; Liang, B; Li, P; Li, H; Yi, S; Liu, H; Cui, X; Wu, Z; Xie, J; Jia, L; Wang, L; Hao, R; Jin, H; Wang, Y; Sun, Y; Song, H

2015-03-01

We identified 2912 Shigella isolates from diarrhoeal patients in China during 2003-2013. The most common species was Shigella flexneri (55.3%), followed by Shigella sonnei (44.1%); however, S. sonnei is becoming increasingly prevalent. Among the S. flexneri isolates, serotypes 2a and X variant (-:7,8, E1037) were the two most prevalent serotypes, and serologically atypical isolates were also commonly identified. Overall, S. sonnei, S. flexneri 2a and S. flexneri X variant (-:7,8, E1037) accounted for 76.1% of all Shigella isolates, and their prevalence increased from 54.0% during 2003-2004 to 84.1% during 2011-2013. A change was observed in the serotype distribution of Shigella in China during this period, and we propose an ideal strategy to inform the development of a broadly effective Shigella vaccine candidate. Copyright © 2014 European Society of Clinical Microbiology and Infectious Diseases. Published by Elsevier Ltd. All rights reserved.

Molecular diagnosis of non-serotypeable Shigella spp.: problems and prospects.

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Muthuirulandi Sethuvel, Dhiviya Prabaa; Devanga Ragupathi, Naveen Kumar; Anandan, Shalini; Walia, Kamini; Veeraraghavan, Balaji

2017-02-01

It is not always possible to identify Shigella serogroups/serotypes by biochemical properties alone. Specific identification requires serotyping. Occasionally, isolates that resemble Shigella spp. biochemically, but are non-agglutinable with available antisera, have been observed. Several mechanisms have been reported to limit the efficiency of the serotyping assay. Serotype conversion is a major mechanism in Shigella spp. to escape protective host immune responses. This easy conversion through significant modification of the O-antigen backbone results in different serotypes, which makes laboratory identification difficult. Furthermore, members of the family Enterobacteriaceae are closely related and there is antigenic cross-over (intra- and inter-specific cross-reaction) which affects the agglutination reaction. The performance of the available methods for identification of non-serotypeable Shigella is discussed here, and reveals them to be non-reliable. This shows a need for an alternative method for identification and typing of Shigella spp.

Sorption Kinetics of Escherichia coli and Salmonella sp on Two Soil Layers Associated with a Groundwater Table in Yaounde, Cameroon (Central Africa

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Norbert Kemka

2005-12-01

Full Text Available A laboratory study has been carried out on two soil layers (HX and HY located above a groundwater table in Yaounde, Cameroon (Central Africa. The main purpose of this study was to assess the retention potential or sorption kinetics of Escherichia coli and Salmonella sp. on these soil layers. For both soil layers, bacterial sorption on soil particles occurred rapidly during the first 30 minutes of incubation of bacteria and soil particles in aqueous media, and increased gradually with incubation time up to 300 min. In some cases, adsorption rates fluctuated after 30 min of incubation, probably due to bacterial cell sorption to and de-sorption from soil particles. Using Freundlich isotherms, it was noted that adsorption coefficient related to adsorption capacity varied from 19 to 4026 E. coli.mg-1 of soil, and from 506 to 847 Salmonella sp.mg-1 of soil. For both bacterial species, the adsorption coefficient of layer HY (located in close proximity of the water table was greater than that of HX (located above layer HY and seemed to positively correlate with the pH values and N/P ratios, and to negatively correlate with the values of C/N and C/P ratios. The linearity coefficient related to adsorption intensity varied from 0.5841 to 1.0023 for E. coli, and from 0.7068 to 1.5236 for Salmonella sp. The physico-chemical characteristics of soil particles seemed to influence the sorption kinetics of bacteria on soil.

Quantitative microbial risk assessment for Escherichia coli O157:H7, salmonella, and Listeria monocytogenes in leafy green vegetables consumed at salad bars.

Science.gov (United States)

Franz, E; Tromp, S O; Rijgersberg, H; van der Fels-Klerx, H J

2010-02-01

Fresh vegetables are increasingly recognized as a source of foodborne outbreaks in many parts of the world. The purpose of this study was to conduct a quantitative microbial risk assessment for Escherichia coli O157:H7, Salmonella, and Listeria monocytogenes infection from consumption of leafy green vegetables in salad from salad bars in The Netherlands. Pathogen growth was modeled in Aladin (Agro Logistics Analysis and Design Instrument) using time-temperature profiles in the chilled supply chain and one particular restaurant with a salad bar. A second-order Monte Carlo risk assessment model was constructed (using @Risk) to estimate the public health effects. The temperature in the studied cold chain was well controlled below 5 degrees C. Growth of E. coli O157:H7 and Salmonella was minimal (17 and 15%, respectively). Growth of L. monocytogenes was considerably greater (194%). Based on first-order Monte Carlo simulations, the average number of cases per year in The Netherlands associated the consumption leafy greens in salads from salad bars was 166, 187, and 0.3 for E. coli O157:H7, Salmonella, and L. monocytogenes, respectively. The ranges of the average number of annual cases as estimated by second-order Monte Carlo simulation (with prevalence and number of visitors as uncertain variables) were 42 to 551 for E. coli O157:H7, 81 to 281 for Salmonella, and 0.1 to 0.9 for L. monocytogenes. This study included an integration of modeling pathogen growth in the supply chain of fresh leafy vegetables destined for restaurant salad bars using software designed to model and design logistics and modeling the public health effects using probabilistic risk assessment software.

Evaluation of aqueous and alcohol-based quaternary ammonium sanitizers for inactivating Salmonella spp., Escherichia coli O157:H7, and Listeria monocytogenes on peanut and pistachio shells.

Science.gov (United States)

McEgan, Rachel; Danyluk, Michelle D

2015-05-01

This study evaluated the efficacy of aqueous (aQUAT) and isopropyl alcohol-based quaternary ammonium (ipQUAT) sanitizers for reducing Salmonella spp., Escherichia coli O157:H7, or Listeria monocytogenes populations on peanut and pistachio shell pieces. Inoculated nutshells were mixed with QUAT sanitizers, water, or 70% ethanol and enumerated immediately or after incubation at 30 °C for 48 h. None of the treatments had any immediate effect on Salmonella or E. coli O157:H7 populations on the peanut or pistachio shells. L. monocytogenes populations declined immediately on the peanut and pistachio shells treated with aQUAT or ipQUAT. After incubation, Salmonella and E. coli O157:H7 populations increased significantly on the water- or aQUAT-treated peanut and pistachio shells. L. monocytogenes populations also increased significantly on the water- or aQUAT-treated peanut shells, but levels did not change on the water-treated pistachio shells and levels were just above the limit of detection on the aQUAT-treated pistachio shells. After treatment with ipQUAT and 48-h incubation, Salmonella and E. coli O157:H7 populations decreased to or below the limit of detection on both shell types; L. monocytogenes populations remained at or below the limit of detection on both shell types. Copyright © 2014 Elsevier Ltd. All rights reserved.

How Shigella Utilizes Ca(2+) Jagged Edge Signals during Invasion of Epithelial Cells.

Science.gov (United States)

Bonnet, Mariette; Tran Van Nhieu, Guy

2016-01-01

Shigella, the causative agent of bacillary dysentery invades intestinal epithelial cells using a type III secretion system (T3SS). Through the injection of type III effectors, Shigella manipulates the actin cytoskeleton to induce its internalization in epithelial cells. At early invasion stages, Shigella induces atypical Ca(2+) responses confined at entry sites allowing local cytoskeletal remodeling for bacteria engulfment. Global Ca(2+) increase in the cell triggers the opening of connexin hemichannels at the plasma membrane that releases ATP in the extracellular milieu, favoring Shigella invasion and spreading through purinergic receptor signaling. During intracellular replication, Shigella regulates inflammatory and death pathways to disseminate within the epithelium. At later stages of infection, Shigella downregulates hemichannel opening and the release of extracellular ATP to dampen inflammatory signals. To avoid premature cell death, Shigella activates cell survival by upregulating the PI3K/Akt pathway and downregulating the levels of p53. Furthermore, Shigella interferes with pro-apoptotic caspases, and orients infected cells toward a slow necrotic cell death linked to mitochondrial Ca(2+) overload. In this review, we will focus on the role of Ca(2+) responses and their regulation by Shigella during the different stages of bacterial infection.

Cloning and properties of the Salmonella typhimurium tricarboxylate transport operon in Escherichia coli

International Nuclear Information System (INIS)

Widenhorn, K.A.; Boos, W.; Somers, J.M.; Kay, W.W.

1988-01-01

The tricarboxylate transport operon (tctI) was cloned in Escherichia coli as a 12-kilobase (kb) fragment from an EcoRI library of the Salmonella typhimurium chromosome in λgtWES. It was further subcloned as a 12-kb fragment into pACYC184 and as an 8-kb fragment into pBR322. By insertional mutagenesis mediated by λTn5, restriction mapping, and phenotypic testing, the tctI operon was localized to a 4.5-kb region. The tctC gene which encodes a periplasmic binding protein (C-protein) was located near the center of the insert. E. coli/tctI clones on either multicopy or single-copy vectors grew on the same tricarboxylates as S. typhimurium, although unusually long growth lags were observed. E. coli/tctI clones exhibited similar [ 14 C] fluorocitrate transport kinetics to those of S. typhimurium, whereas E. coli alone was virtually impermeable to [ 14 C] fluorocitrate. The periplasmic C proteins (C1 and C2 isoelectric forms) were produced in prodigious quantities from the cloned strains. Motile E. coli/tctI clones were not chemotactic toward citrate, whereas tctI deletion mutants of S. typhimurium were. Taken together, these observations indicate that tctI is not an operon involved in chemotaxis

Decontamination of tomato, red cabbage, carrot, fresh parsley and fresh green onion inoculated with Shigella sonnei and Shigella flexneri by some Essential oils (in vitro condition

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Farhang Aliakbari

2014-06-01

Full Text Available Background: Essential oils and their major constituents are useful sources of antimicrobial compounds. There are a few reports on the decontamination and antimicrobial activity of essential oils towards Shigella spp.Materials and Methods: In this study, the antimicrobial and decontamination potentials of essential oils at different concentrations, belonging to plants such as Thymus vulgaris, Saturiea hortensis, Mentha polegium, Cuminum cyminum, Lavandula officinalis and Mentha viridis L. (spearmint, towards Shigella sonnei and Shigella  flexneri were investigated. The disk diffusion method demonstrated the antimicrobial potential of the essential oils.Results: The ability of essential oils to decontaminate vegetables such as, tomato, red cabbage, carrot, fresh parsley and fresh green onion that were previously inoculated with Shigella spp. was determined. Inhibitory effects of essential oils towards Shigella spp. were noted in the disk diffusion method. There was a reduction in Shigella population following inoculation of cultures with 0.5% and 0.1% (v/v essential oils.Conclusion: This study confirmed that essential oils have the potential to be used for decontamination of vegetables.

Negligible import of enteric pathogens by newly-arrived asylum seekers and no impact on incidence of notified Salmonella and Shigella infections and outbreaks in Rhineland-Palatinate, Germany, January 2015 to May 2016.

Science.gov (United States)

Ehlkes, Lutz; George, Maja; Knautz, Donald; Burckhardt, Florian; Jahn, Klaus; Vogt, Manfred; Zanger, Philipp

2018-05-01

IntroductionThe 2015 refugee crisis raised concerns about an import of infectious diseases affecting the German population. Aims: To evaluate public and individual health benefits of stool screening, and explore whether importation of enteric pathogens by newly-arrived asylum seekers impacts on the host population. Methods : We used data from mandatory stool screening to determine the overall, age, sex, and country-specific prevalence of enteric bacteria and helminths. We used surveillance data to assess whether the number of incoming asylum seekers influenced notifications of salmonellosis and shigellosis in Rhineland-Palatinate. Results : Salmonella were found in 0.2% (95% confidence interval (CI) 0.2-0.3%) of 23,410 samples collected from January 2015 to May 2016. Prevalence was highest in children under 5 years (0.8%; 95% CI: 0.5-1.3%). No Shigella or invasive Salmonella spp. were detected. In a subset of 14,511 samples, the prevalence of helminth infestation was 2.4% (95% CI: 2.1-2.6%), with highest proportions detected in adolescents (4.6%; 95% CI 3.8-5.4%) and among Eritreans (9.3%; 95% CI: 7.0-12.0%); in the latter particularly Schistosoma mansoni and Taenia spp. The increase in asylum applications did not increase notifications of salmonellosis and shigellosis. No transmission from asylum seekers to German residents was notified. Conclusion : Public health risk associated with imported enteric pathogens is very low overall. Addressing individual and public health risks, we recommend replacing stool screening of all newly-arrived asylum seekers by a targeted approach, with target groups and approaches being adapted if necessary. Target groups supported by our data are children, adolescents, and Eritreans.

Monitoring of the dissemination of Salmonella in the chicken Frankfurt-sausage productionline of a sausage factory in the state of São Paulo, Brazil

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Alexandre de Freitas Luiz

2004-08-01

Full Text Available Poultry meat and its derivatives are among the foodstuffs considered by environmental health authorities to present the highest risks to the public. A total of 185 samples were collected in five monthly batches, from different processing stages in a sausage plant that uses mechanically-deboned chicken meat (MDCM, and testedfor the presence of Salmonella. Enrichment was carried out in both Kauffman's tetrathionate broth and Rappaport-Vassiliadis broth and isolation on Salmonella-Shigella agar and brilliant-green agar. Live Salmonella bacteria were isolated from six samples of the raw meat and from the emulsion, in batches three, four, and five, but not from any sample in batches one or two. The six isolated strains were all classified as Salmonella Albany, which has not previously been reported in MDCM. Of the two enrichment broths, Rappaport-Vassiliadis gave the better results. The pattern of contamination suggests a probable common source, given that a new supplier was used in the third, fourth, and fifth months. It was also shown that the industrial cooking was effective in preventing Salmonella surviving in the final product.

Virulotyping of Shigella spp. isolated from pediatric patients in Tehran, Iran.

Science.gov (United States)

Ranjbar, Reza; Bolandian, Masomeh; Behzadi, Payam

2017-03-01

Shigellosis is a considerable infectious disease with high morbidity and mortality among children worldwide. In this survey the prevalence of four important virulence genes including ial, ipaH, set1A, and set1B were investigated among Shigella strains and the related gene profiles identified in the present investigation, stool specimens were collected from children who were referred to two hospitals in Tehran, Iran. The samples were collected during 3 years (2008-2010) from children who were suspected to shigellosis. Shigella spp. were identified throughout microbiological and serological tests and then subjected to PCR for virulotyping. Shigella sonnei was ranking first (65.5%) followed by Shigella flexneri (25.9%), Shigella boydii (6.9%), and Shigella dysenteriae (1.7%). The ial gene was the most frequent virulence gene among isolated bacterial strains and was followed by ipaH, set1B, and set1A. S. flexneri possessed all of the studied virulence genes (ial 65.51%, ipaH 58.62%, set1A 12.07%, and set1B 22.41%). Moreover, the pattern of virulence gene profiles including ial, ial-ipaH, ial-ipaH-set1B, and ial-ipaH-set1B-set1A was identified for isolated Shigella spp. strains. The pattern of virulence genes is changed in isolated strains of Shigella in this study. So, the ial gene is placed first and the ipaH in second.

Survival of enteric pathogens in common beverages: an in vitro study.

Science.gov (United States)

Sheth, N K; Wisniewski, T R; Franson, T R

1988-06-01

This in vitro study was undertaken to determine the potential for survival of enteric pathogens in common drinking beverages. Three carbonated soft drinks, two alcoholic beverages, skim milk, and water were inoculated with Salmonella, Shigella, and enterotoxigenic Escherichia coli, and quantitative counts were performed over 2 days. Our studies showed poorest survival of all three organisms in wine, and greatest growth in milk and water. Beer and cola allowed survival of small numbers of Salmonella and E. coli at 48 h, whereas sour mix and diet cola were sterile by 48 h. Survival features may correlate with pH of the beverages. These observations may be useful in guiding travellers for appropriate beverage consumption while visiting areas endemic for "traveller's diarrhea."

Inactivation of Escherichia coli, Listeria monocytogenes, and Salmonella Enteritidis by Cymbopogon citratus D.C. Stapf. Essential Oil in Pineapple Juice.

Science.gov (United States)

Leite, Caroline Junqueira Barcellos; de Sousa, Jossana Pereira; Medeiros, José Alberto da Costa; da Conceição, Maria Lúcia; dos Santos Falcão-Silva, Vivyanne; de Souza, Evandro Leite

2016-02-01

In the present study, the efficacy of Cymbopogon citratus D.C. Stapf. essential oil (CCEO) to provoke a 5-log CFU/ml (5-log) inactivation in a mixed composite of Escherichia coli, Listeria monocytogenes, and Salmonella enterica serovar Enteritidis in pineapple (Ananas comosus (L.) Merril) juice (4°C) was assessed. Moreover, the effects of CCEO on the physicochemical and sensory quality parameters of pineapple juice were evaluated. The MIC of CCEO was 5 μl/ml against the composite mix examined. For L. monocytogenes and E. coli inoculated in juice containing CCEO (5, 2.5, and 1.25 μl/ml), a ≥5-log reduction was detected after 15 min of exposure. This same result was obtained for Salmonella Enteritidis incubated alone in pineapple juice containing CCEO at 5 and 2.5 μl/ml. Overall, Salmonella Enteritidis was the most tolerant and L. monocytogenes was the most sensitive to CCEO. The physicochemical properties (pH, titratable acidic [citric acid per 100 g], and soluble solids) of pineapple juice containing CCEO (2.5 and 1.25 μl/ml) were maintained. Juice containing CCEO (2.5 and 1.25 μl/ml) exhibited similar scores for odor, appearance, and viscosity compared with juice without CCEO. However, unsatisfactory changes in taste and aftertaste were observed in juices containing CCEO. These results suggest that CCEO could be used as an alternative antimicrobial compound to ensure the safety of pineapple juice, although CCEO at the tested concentrations negatively impacted its taste. Therefore, further studies are needed to determine the balance between microbial safety and taste acceptability of pineapple juice containing CCEO.

Efecto de la exposición a la luz ultravioleta uv-c en la viabilidad de especies de Eschericha coli y Salmonella typhimurium

OpenAIRE

Oviedo, Dumas; Rojas, Jesús María; Borda, Ricardo Alberto; Durango, Mónica María

2013-01-01

Introduction. The germicidal effect UV-C light has is regarded as an effective tool to inactivate and eliminate harmful contaminating agents, such as Escherichia coli and Salmonella typhimurium. Objective. evaluating the effectiveness of UV-C light for reducing Escherichia coli and Salmonella typhimorium populations from cultures that had the microorganisms, combining factors like concentration, time and distance. Methodology. A UV-C lamp, with a 254 nm and 8 ...

Arginine-dependent acid resistance in Salmonella enterica serovar Typhimurium

NARCIS (Netherlands)

Kieboom, J.; Abee, T.

2006-01-01

Salmonella enterica serovar Typhimurium does not survive a pH 2.5 acid challenge under conditions similar to those used for Escherichia coli (J. W. Foster, Nat. Rev. Microbiol. 2:898-907, 2004). Here, we provide evidence that S. enterica serovar Typhimurium can display arginine-dependent acid

Aerobic bacterial microflora of Broad-snouted caiman (Caiman latirostris oral cavity and cloaca, originating from Parque Zoológico Arruda Câmara, Paraíba, Brazil Microflora bacteriana aeróbica da cavidade oral e cloaca de jacaré-de-papo-amarelo (Caiman latirostris procedentes do Zoológico de João Pessoa, PB, Brasil

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J.S.A. Silva

2009-03-01

Full Text Available The objective of this study was to isolate and identify the aerobic bacterial microflora from the oral cavity mucosa and cloaca's samples, collected from Broad-snouted caiman (Caiman latirostris, born and bred in captivity at Parque Zoológico Arruda Câmara, João Pessoa, Paraíba, Brazil. The most common bacteria were Staphylococcus sp. (14.74%, Corynebacterium sp. (13.68%, Escherichia coli (13.68% and Shigella sp. (11.58%, and the less common were Citrobacter sp. (1.05%, Klebsiella pneumoniae (1.05% and Salmonella sp. (1.05%.This emphasizes the importance of these microorganisms' participation in infectious processes (sepsis and injuries caused by crocodilians.O presente estudo teve como objetivo isolar e identificar a microflora bacteriana aeróbica presente na mucosa da cavidade oral e da cloaca de exemplares de jacarés-de-papo-amarelo (Caiman latirostris nascidos e criados em cativeiro no Parque Zoológico Arruda Câmara, localizado na cidade de João Pessoa - PB. As bactérias mais freqüentes foram Staphylococcus sp. (14,74%, Corynebacterium sp.(13,68%, Escherichia coli (13,68% e Shigella sp. (11,58%, e as menos prevalentes foram Citrobacter sp.(1,05%, Klebsiella pneumoniae (1,05% e Salmonella sp. (1,05%. Ressalta-se a importância da participação desses microrganismos em processos infecciosos (septicemias e em feridas provocadas por crocodilianos.

Analysis of plasmid genes by phylogenetic profiling and visualization of homology relationships using Blast2Network

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Bazzicalupo Marco

2008-12-01

Full Text Available Abstract Background Phylogenetic methods are well-established bioinformatic tools for sequence analysis, allowing to describe the non-independencies of sequences because of their common ancestor. However, the evolutionary profiles of bacterial genes are often complicated by hidden paralogy and extensive and/or (multiple horizontal gene transfer (HGT events which make bifurcating trees often inappropriate. In this context, plasmid sequences are paradigms of network-like relationships characterizing the evolution of prokaryotes. Actually, they can be transferred among different organisms allowing the dissemination of novel functions, thus playing a pivotal role in prokaryotic evolution. However, the study of their evolutionary dynamics is complicated by the absence of universally shared genes, a prerequisite for phylogenetic analyses. Results To overcome such limitations we developed a bioinformatic package, named Blast2Network (B2N, allowing the automatic phylogenetic profiling and the visualization of homology relationships in a large number of plasmid sequences. The software was applied to the study of 47 completely sequenced plasmids coming from Escherichia, Salmonella and Shigella spps. Conclusion The tools implemented by B2N allow to describe and visualize in a new way some of the evolutionary features of plasmid molecules of Enterobacteriaceae; in particular it helped to shed some light on the complex history of Escherichia, Salmonella and Shigella plasmids and to focus on possible roles of unannotated proteins. The proposed methodology is general enough to be used for comparative genomic analyses of bacteria.

Novo meio seletivo-indicador para detecção de Aeromonas e Plesiomonas: ágar UNISC New selective indicator medium for detection of Aeromonas and Plesiomonas: UNISC agar

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Marion Pereira da Rocha

2008-08-01

Full Text Available Avaliou-se um novo meio seletivo-indicador (ágar UNISC para o isolamento de enteropatógenos clássicos e Aeromonas e Plesiomonas shigelloides. A capacidade de fermentação da xilose é indicada pela coloração amarela (fermentadores ou azul (não fermentadores que, aliada à prova da oxidase, constitui-se em indicador para a detecção de Aeromonas spp e Plesiomonas shigelloides. A produtividade e seletividade, avaliadas pelos índice de contagem absoluta e índice de contagem relativa indicam-no como uma alternativa aos coprocultivos clássicos porque permite, num só meio, o isolamento de Escherichia coli, Shigella spp, Salmonella spp, bem como, Aeromonas spp e Plesiomonas shigelloides, favorecendo o diagnóstico laboratorial das gastroenterites.We evaluated a new selective indicator medium (UNISC Agar for isolation of classical enteropathogens, Aeromonas spp and Plesiomonas shigelloides. The xylose fermentation capacity is indicated by a yellow color (fermenting agents or blue (no fermenting agent. This, together with the oxidase test, establishes it as an indicator for detecting Aeromonas and Plesiomonas shigelloides. Its productivity and selectivity, as assessed using the absolute count index and relative count index, indicate it as an alternative to the classical feces culturing media. This is because, in a single medium, it enables isolation of Escherichia coli, Shigella spp and Salmonella spp, in addition to Aeromonas and Plesiomonas shigelloides, thereby favoring the laboratory diagnosis of gastroenteritis.

Modulation of Caenorhabditis elegans immune response and modification of Shigella endotoxin upon interaction.

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Kesika, Periyanaina; Prasanth, Mani Iyer; Balamurugan, Krishnaswamy

2015-04-01

To analyze the pathogenesis at both physiological and molecular level using the model organism, Caenorhabditis elegans at different developmental stages in response to Shigella spp. and its pathogen associated molecular patterns such as lipopolysaccharide. The solid plate and liquid culture-based infection assays revealed that Shigella spp. infects C. elegans and had an impact on the brood size and pharyngeal pumping rate. LPS of Shigella spp. was toxic to C. elegans. qPCR analysis revealed that host innate immune genes have been modulated upon Shigella spp. infections and its LPS challenges. Non-destructive analysis was performed to kinetically assess the alterations in LPS during interaction of Shigella spp. with C. elegans. The modulation of innate immune genes attributed the surrendering of host immune system to Shigella spp. by favoring the infection. LPS appeared to have a major role in Shigella-mediated pathogenesis and Shigella employs a tactic behavior of modifying its LPS content to escape from the recognition of host immune system. © 2014 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

Efficacy of a Blend of Sulfuric Acid and Sodium Sulfate against Shiga Toxin-Producing Escherichia coli, Salmonella, and Nonpathogenic Escherichia coli Biotype I on Inoculated Prerigor Beef Surface Tissue.

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Scott-Bullard, Britteny R; Geornaras, Ifigenia; Delmore, Robert J; Woerner, Dale R; Reagan, James O; Morgan, J Bred; Belk, Keith E

2017-12-01

A study was conducted to investigate the efficacy of a sulfuric acid-sodium sulfate blend (SSS) against Escherichia coli O157:H7, non-O157 Shiga toxin-producing E. coli (STEC), Salmonella, and nonpathogenic E. coli biotype I on prerigor beef surface tissue. The suitability of using the nonpathogenic E. coli as a surrogate for in-plant validation studies was also determined by comparing the data obtained for the nonpathogenic inoculum with those for the pathogenic inocula. Prerigor beef tissue samples (10 by 10 cm) were inoculated (ca. 6 log CFU/cm 2 ) on the adipose side in a laboratory-scale spray cabinet with multistrain mixtures of E. coli O157:H7 (5 strains), non-O157 STEC (12 strains), Salmonella (6 strains), or E. coli biotype I (5 strains). Treatment parameters evaluated were two SSS pH values (1.5 and 1.0) and two spray application pressures (13 and 22 lb/in 2 ). Untreated inoculated beef tissue samples served as controls for initial bacterial populations. Overall, the SSS treatments lowered inoculated (6.1 to 6.4 log CFU/cm 2 ) bacterial populations by 0.6 to 1.5 log CFU/cm 2 (P SSS was applied to samples inoculated with any of the tested E. coli inocula; however, solution pH did have a significant effect (P SSS was applied to samples inoculated with Salmonella. Results indicated that the response of the nonpathogenic E. coli inoculum to the SSS treatments was similar (P ≥ 0.05) to that of the pathogenic inocula tested, making the E. coli biotype I strains viable surrogate organisms for in-plant validation of SSS efficacy on beef. The application of SSS at the tested parameters to prerigor beef surface tissue may be an effective intervention for controlling pathogens in a commercial beef harvest process.

How Shigella Utilizes Ca2+ Jagged Edge Signals during Invasion of Epithelial Cells

Science.gov (United States)

Bonnet, Mariette; Tran Van Nhieu, Guy

2016-01-01

Shigella, the causative agent of bacillary dysentery invades intestinal epithelial cells using a type III secretion system (T3SS). Through the injection of type III effectors, Shigella manipulates the actin cytoskeleton to induce its internalization in epithelial cells. At early invasion stages, Shigella induces atypical Ca2+ responses confined at entry sites allowing local cytoskeletal remodeling for bacteria engulfment. Global Ca2+ increase in the cell triggers the opening of connexin hemichannels at the plasma membrane that releases ATP in the extracellular milieu, favoring Shigella invasion and spreading through purinergic receptor signaling. During intracellular replication, Shigella regulates inflammatory and death pathways to disseminate within the epithelium. At later stages of infection, Shigella downregulates hemichannel opening and the release of extracellular ATP to dampen inflammatory signals. To avoid premature cell death, Shigella activates cell survival by upregulating the PI3K/Akt pathway and downregulating the levels of p53. Furthermore, Shigella interferes with pro-apoptotic caspases, and orients infected cells toward a slow necrotic cell death linked to mitochondrial Ca2+ overload. In this review, we will focus on the role of Ca2+ responses and their regulation by Shigella during the different stages of bacterial infection. PMID:26904514

How Shigella utilizes Ca2+ jagged edge signals during invasion of epithelial cells

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Mariette eBonnet

2016-02-01

Full Text Available Shigella, the causative agent of bacillary dysentery invades intestinal epithelial cells using a type III secretion system (T3SS. Through the injection of type III effectors, Shigella manipulates the actin cytoskeleton to induce its internalization in epithelial cells. At early invasion stages, Shigella induces atypical Ca2+ responses confined at entry sites allowing local cytoskeletal remodeling for bacteria engulfment. Global Ca2+ increase in the cell triggers the opening of connexin hemichannels at the plasma membrane that releases ATP in the extracellular milieu, favoring Shigella invasion and spreading through purinergic receptor signaling. During intracellular replication, Shigella regulates inflammatory and death pathways to disseminate within the epithelium. At later stages of infection, Shigella downregulates hemichannel opening and the release of extracellular ATP to dampen inflammatory signals. To avoid premature cell death, Shigella activates cell survival by upregulating the PI3K/Akt pathway and downregulating the levels of p53. Furthermore, Shigella interferes with pro-apoptotic caspases, and orients infected cells towards a slow necrotic cell death linked to mitochondrial Ca2+ overload. In this review, we will focus on the role of Ca2+ responses and their regulation by Shigella during the different stages of bacterial infection.

Brote causado por Escherichia coli en Chalco, México Outbreak caused by Escherichia coli in Chalco, México

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Iliana Alejandra Cortés-Ortiz

2002-07-01

Full Text Available Objetivo. Identificar el agente causal del brote de diarrea asociado con el desbordamiento del canal de aguas negras en Chalco. Material y métodos. Estudio retrospectivo y transversal, efectuado en el Instituto de Diagnóstico y Referencia Epidemiológicos (InDRE, de la Secretaría de Salud, con 1 550 hisopos rectales para el aislamiento e identificación bioquímica de V. cholerae y enterobacterias, obtenidos de la población del Valle de Chalco, que presentó diarrea y vómito durante el desastre natural acontecido el 31 de mayo de 2000. El análisis de los resultados se efectuó por la diferencia entre las proporciones de dos poblaciones (prueba de Ji cuadrada. Las cepas de E. coli se hibridaron por "colony blot" para los grupos ETEC, EIEC, EPEC y EHEC. Resultados. El 0.45% correspondió a Salmonella: S. agona, S. infantis, S. enteritidis, S. muenchen, S. typhimurium; 0.06% a Shigella flexneri 3a, y 76.6% a E. coli: 62.2% a ETEC (44.6 % con LT, 11.2% con ST, y 44.1% con ambas sondas, 0.84% a EIEC (sonda ial, 0.84% a EPEC (sonda bundle-forming pilus BFP, 0.08% a E. coli enterohemorrágica no-O157:H7 (sonda pCVD419, y 36.02% no hibridó. No se encontró asociación entre E. coli patógena con la edad y género. Conclusiones. Escherichia coli podría ser responsable del brote de diarrea. Es importante conocer el agente etiológico del brote para encaminar las estrategias en el estudio y control sanitario del mismo.Objective. To identify the etiologic agent responsible for a disease outbreak following an overflow of sewage water in Valle de Chalco, Mexico. Material and Methods. A retrospective cross-sectional study was carried out. Rectal samples were collected from the population of Chalco valley, who suffered from diarrhea and vomiting during a natural disaster that took place on May 31, 2000. The Instituto de Diagnóstico y Referencia Epidemiológicos (Epidemic Reference and Diagnosis Institute, InDRE, Ministry of Health, received 1521 rectal

Isolation of OmpA gene from Salmonella typhimurium and ...

African Journals Online (AJOL)

Isolation of OmpA gene from Salmonella typhimurium and transformation into alfalfa in order to develop an edible plant based vaccine. ... The recombinant OmpA was expressed in Escherichia coli TG1. The new construct was used to transform the Agrobacterium tumefaciens Strain LBA4404 before plant transformation.

A clinically parameterized mathematical model of Shigella immunity to inform vaccine design.

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Courtney L Davis

Full Text Available We refine and clinically parameterize a mathematical model of the humoral immune response against Shigella, a diarrheal bacteria that infects 80-165 million people and kills an estimated 600,000 people worldwide each year. Using Latin hypercube sampling and Monte Carlo simulations for parameter estimation, we fit our model to human immune data from two Shigella EcSf2a-2 vaccine trials and a rechallenge study in which antibody and B-cell responses against Shigella's lipopolysaccharide (LPS and O-membrane proteins (OMP were recorded. The clinically grounded model is used to mathematically investigate which key immune mechanisms and bacterial targets confer immunity against Shigella and to predict which humoral immune components should be elicited to create a protective vaccine against Shigella. The model offers insight into why the EcSf2a-2 vaccine had low efficacy and demonstrates that at a group level a humoral immune response induced by EcSf2a-2 vaccine or wild-type challenge against Shigella's LPS or OMP does not appear sufficient for protection. That is, the model predicts an uncontrolled infection of gut epithelial cells that is present across all best-fit model parameterizations when fit to EcSf2a-2 vaccine or wild-type challenge data. Using sensitivity analysis, we explore which model parameter values must be altered to prevent the destructive epithelial invasion by Shigella bacteria and identify four key parameter groups as potential vaccine targets or immune correlates: 1 the rate that Shigella migrates into the lamina propria or epithelium, 2 the rate that memory B cells (BM differentiate into antibody-secreting cells (ASC, 3 the rate at which antibodies are produced by activated ASC, and 4 the Shigella-specific BM carrying capacity. This paper underscores the need for a multifaceted approach in ongoing efforts to design an effective Shigella vaccine.

A clinically parameterized mathematical model of Shigella immunity to inform vaccine design.

Science.gov (United States)

Davis, Courtney L; Wahid, Rezwanul; Toapanta, Franklin R; Simon, Jakub K; Sztein, Marcelo B

2018-01-01

We refine and clinically parameterize a mathematical model of the humoral immune response against Shigella, a diarrheal bacteria that infects 80-165 million people and kills an estimated 600,000 people worldwide each year. Using Latin hypercube sampling and Monte Carlo simulations for parameter estimation, we fit our model to human immune data from two Shigella EcSf2a-2 vaccine trials and a rechallenge study in which antibody and B-cell responses against Shigella's lipopolysaccharide (LPS) and O-membrane proteins (OMP) were recorded. The clinically grounded model is used to mathematically investigate which key immune mechanisms and bacterial targets confer immunity against Shigella and to predict which humoral immune components should be elicited to create a protective vaccine against Shigella. The model offers insight into why the EcSf2a-2 vaccine had low efficacy and demonstrates that at a group level a humoral immune response induced by EcSf2a-2 vaccine or wild-type challenge against Shigella's LPS or OMP does not appear sufficient for protection. That is, the model predicts an uncontrolled infection of gut epithelial cells that is present across all best-fit model parameterizations when fit to EcSf2a-2 vaccine or wild-type challenge data. Using sensitivity analysis, we explore which model parameter values must be altered to prevent the destructive epithelial invasion by Shigella bacteria and identify four key parameter groups as potential vaccine targets or immune correlates: 1) the rate that Shigella migrates into the lamina propria or epithelium, 2) the rate that memory B cells (BM) differentiate into antibody-secreting cells (ASC), 3) the rate at which antibodies are produced by activated ASC, and 4) the Shigella-specific BM carrying capacity. This paper underscores the need for a multifaceted approach in ongoing efforts to design an effective Shigella vaccine.

Potassium transport of Salmonella is important for type III secretion and pathogenesis

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Liu, Yehao; Ho, Katharina Kim; Su, Jing; Gong, Hao; Chang, Alexander C.

2013-01-01

Intracellular cations are essential for the physiology of all living organisms including bacteria. Cations such as potassium ion (K+), sodium ion (Na+) and proton (H+) are involved in nearly all aspects of bacterial growth and survival. K+ is the most abundant cation and its homeostasis in Escherichia coli and Salmonella is regulated by three major K+ transporters: high affinity transporter Kdp and low affinity transporters Kup and Trk. Previous studies have demonstrated the roles of cations and cation transport in the physiology of Escherichia coli; their roles in the virulence and physiology of pathogenic bacteria are not well characterized. We have previously reported that the Salmonella K+ transporter Trk is important for the secretion of effector proteins of the type III secretion system (TTSS) of Salmonella pathogenicity island 1 (SPI-1). Here we further explore the role of Salmonella cation transport in virulence in vitro and pathogenesis in animal models. Impairment of K+ transport through deletion of K+ transporters or exposure to the chemical modulators of cation transport, gramicidin and valinomycin, results in a severe defect in the TTSS of SPI-1, and this defect in the TTSS was not due to a failure to regulate intrabacterial pH or ATP. Our results also show that K+ transporters are critical to the pathogenesis of Salmonella in mice and chicks and are involved in multiple growth and virulence characteristics in vitro, including protein secretion, motility and invasion of epithelial cells. These results suggest that cation transport of the pathogenic bacterium Salmonella, especially K+ transport, contributes to its virulence in addition to previously characterized roles in maintaining homeostasis of bacteria. PMID:23728623

Detection of Escherichia coli O157:H7 and Salmonella in ground beef by a bead-free quantum dot-facilitated isolation method.

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Wang, Luxin; Wu, Chung-Shieh; Fan, Xudong; Mustapha, Azlin

2012-05-01

The aims of this study were to introduce a new immunological bead-free cell detection method using quantum dots (QDs) as reporter markers for foodborne pathogen detection. QDs are nanosized particles with long-term photostability, high quantum yield, broad absorption spectra, and narrow, symmetric emission and high signal-to-noise ratio. The chemical compound [(1-ethyl-3-3-dimethylaminopropyl) carbodiimide hydrochloride] (EDC) and protein A were used as crosslinkers for manufacturing QD-antibody conjugates. To minimize the inhibition of QD fluorescence by the magnetic beads, the beads were removed after the primary pathogen isolation and before fluorescence measurement. Detection signals were increased four-fold after employing the bead-free isolation method. With a 24-h enrichment, the bead-free QD-facilitated detection method was able to detect 10 CFU/g Escherichia coli O157:H7 and Salmonella from artificially contaminated ground beef. To our knowledge, this detection method is the first research that combined a new EDC-protein A QD-labeling technique and bead-free fluorescence measurement to detect E. coli O157:H7 and Salmonella in ground beef. Copyright © 2012 Elsevier B.V. All rights reserved.

E. coli Nissle 1917 Affects Salmonella adhesion to porcine intestinal epithelial cells.

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Peter Schierack

Full Text Available BACKGROUND: The probiotic Escherichia coli strain Nissle 1917 (EcN has been shown to interfere in a human in vitro model with the invasion of several bacterial pathogens into epithelial cells, but the underlying molecular mechanisms are not known. METHODOLOGY/PRINCIPAL FINDINGS: In this study, we investigated the inhibitory effects of EcN on Salmonella Typhimurium invasion of porcine intestinal epithelial cells, focusing on EcN effects on the various stages of Salmonella infection including intracellular and extracellular Salmonella growth rates, virulence gene regulation, and adhesion. We show that EcN affects the initial Salmonella invasion steps by modulating Salmonella virulence gene regulation and Salmonella SiiE-mediated adhesion, but not extra- and intracellular Salmonella growth. However, the inhibitory activity of EcN against Salmonella invasion always correlated with EcN adhesion capacities. EcN mutants defective in the expression of F1C fimbriae and flagellae were less adherent and less inhibitory toward Salmonella invasion. Another E. coli strain expressing F1C fimbriae was also adherent to IPEC-J2 cells, and was similarly inhibitory against Salmonella invasion like EcN. CONCLUSIONS: We propose that EcN affects Salmonella adhesion through secretory components. This mechanism appears to be common to many E. coli strains, with strong adherence being a prerequisite for an effective reduction of SiiE-mediated Salmonella adhesion.

Laboratory based surveillance of travel-related Shigella sonnei and Shigella flexneri in Alberta from 2002 to 2007

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Ferrato Christina

2010-11-01

Full Text Available Abstract Between 2002 and 2007, travel related cases of Shigella sonnei and S. flexneri in Alberta, Canada were acquired from Central America, the Indian subcontinent and North America. Of this group, resistance to ciprofloxacin and nalidixic acid was identified in isolates from patients who had travelled to the Indian subcontinent. This study provides a Canadian perspective to a growing body of literature linking ciprofloxacin and nalidixic acid resistance to travel to the Indian subcontinent. Shigella is a common cause of diarrheal illness in North America with a rate of 2.0 per 100,000 in Canada 1 and a rate of 3.2 per 100,000 in the United States 23. Imported cases of Shigella infections have been reported in developed countries following travel to a foreign or developing country 45 and may be impacted by factors including socio-economic factors 6, food distribution networks 5 and microbiologic factors 7. Across multiple geographic regions, high rates of antimicrobial resistance to multiple agents (e.g. sulfonamides, tetracycline, chloramphenicol, ampicillin, and trimethoprim-sulfamethoxazole have limited the choices for empiric antimicrobial therapy required to manage Shigella infections and reduce fecal excretion of the bacteria 8910 with descriptions of shifting species dominance and changes in antimicrobial susceptibility 1011. Generally, Shigella flexneri and Shigella sonnei are the dominant species and are heavily impacted by changes in antimicrobial susceptibility 1213. This study identifies the global regions associated with travel-related cases of S. flexneri and S. sonnei in Alberta, Canada and compares antibiotic resistance patterns of these isolates for 2002 to 2007 inclusive. Specimens collected 2002-2007 (inclusive from S. flexneri and S. sonnei infections in Alberta, Canada were included for study. Data collected at time of specimen submission included: date of specimen collection, outbreak association if present, travel

Template synthesis, characterization and antimicrobial activity of the complex combinations of some transitional metals with isonicotinoylhydrazone -2,4,6- trimethylbenzaldehyde

International Nuclear Information System (INIS)

Mitu, L.ITU; Imran, M.

2011-01-01

The complexes of Cu(II), Ni(II), Co(II), Mn(II), and Zn(II) with isonicotinoyl-hydrazone -2,4,6-trimethylbenzaldehyde (INHTB) are reported. The complexes have been characterized by analytical data, IR, UV-Vis, NMR spectra, magnetic susceptibility, thermal analysis and for the Cu(II) complex the ESR spectrum has been recorded. The anti-bacterial activity of these complexes were investigated against Staphylococcus aureus, Escherichia coli, Pseudomonas aeruginosa, Salmonella enteritidis, Shigella flexneri bacteria. The INHTB ligand is coordinated at the metallic ions by oxygen amide (O=C) and the azomethine nitrogen. (author)

Risk factors and outcome of Shigella encephalopathy in Bangladeshi children.

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Farzana Afroze

2017-04-01

Full Text Available Although, Shigella encephalopathy, a serious extra-intestinal complication of shigellosis, significantly increases the risks of death, data are very limited on predicting factors particularly related to electrolyte profiles in children below five years of age with Shigella encephalopathy. Our objective was to determine the clinical as well as laboratory predicting factors and outcome of children with Shigella encephalopathy.In this unmatched case-control design, children aged 2-59 months having a positive stool culture for Shigella and who had their serum electrolytes been done from July 2012 to June 2015 were studied. Children with Shigella encephalopathy, defined as having abnormal mentation, constituted the cases, and those without encephalopathy constituted the controls. During the study period, we identified a total of 541 children less than five years of age, who had Shigella in their stool culture. Only 139 children fulfilled the study criteria and among them 69 were cases and 70 were controls. The cases more often had fatal outcome compared to the controls (7% vs. 0%, P = 0.02. In logistic regression analysis, the cases were independently associated with shorter duration (1.2 ± 0.4 days of diarrhea prior to admission, dehydrating diarrhea, sepsis and hyponatremia (p<0.05 for all. Among 139 Shigella isolates, S. flexneri (88/139, 63% and S. sonnei(34/139, 24% were the dominant species. S. dysenteriae was not isolated throughout the study period. S.sonnei was more frequently isolated from the cases (24/69, 35% than the controls (10/70, 14%, whereas the isolation of S. flexneri was comparable between the groups (40/69, 58% vs 48/70, 69%. A total of 94 (67.6% isolates were resistant to trimethoprim-sulphamethoxazole, 84 (60.4% to ciprofloxacin, 66/138 (48% to ampicillin, 5 (3.5% to ceftriaxone, 17 (12.2% to mecillinum and 35 (25% to azithromycin.The case-fatality-rate was significantly higher among the children with Shigella encephalopathy

Shigella sonnei bacteremia in an elderly diabetic patient.

Science.gov (United States)

Dronda, F; Parras, F; Martínez, J L; Baquero, F

1988-06-01

A case of Shigella sonnei bacteremia in a 65-year-old patient suffering from diabetes mellitus is discussed. The isolated strain had plasmid-mediated serum resistance and excreted aerobactin. The presence of diabetes mellitus as an underlying disease and the production by the Shigella sonnei isolate of factors related to bacterial invasion may have contributed to the bacteremic episode.

Cloning and Expression of Fusion Genes of Domain A-1 Protective Antigen of Bacillus Anthracis and Shigella Enterotoxin B Subunit (Stxb In E. Coil

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AH ahmadi

2015-02-01

Conclusion: The findings of the current study revealed that this antigen can be raised as an anti-cancer and recombinant vaccine candidate against types of Shigella, Escherichia coli and Bacillus anthracis which can be due to such factors as identification of antigen(PA by antibody PA20, its apoptosis induction properties, property of immunogenicity, adjuvant and delivery of STxB protein and high expression levels of Gb3 in human cancer cells.

Bioinformatics analyses of Shigella CRISPR structure and spacer classification.

Science.gov (United States)

Wang, Pengfei; Zhang, Bing; Duan, Guangcai; Wang, Yingfang; Hong, Lijuan; Wang, Linlin; Guo, Xiangjiao; Xi, Yuanlin; Yang, Haiyan

2016-03-01

Clustered regularly interspaced short palindromic repeats (CRISPR) are inheritable genetic elements of a variety of archaea and bacteria and indicative of the bacterial ecological adaptation, conferring acquired immunity against invading foreign nucleic acids. Shigella is an important pathogen for anthroponosis. This study aimed to analyze the features of Shigella CRISPR structure and classify the spacers through bioinformatics approach. Among 107 Shigella, 434 CRISPR structure loci were identified with two to seven loci in different strains. CRISPR-Q1, CRISPR-Q4 and CRISPR-Q5 were widely distributed in Shigella strains. Comparison of the first and last repeats of CRISPR1, CRISPR2 and CRISPR3 revealed several base variants and different stem-loop structures. A total of 259 cas genes were found among these 107 Shigella strains. The cas gene deletions were discovered in 88 strains. However, there is one strain that does not contain cas gene. Intact clusters of cas genes were found in 19 strains. From comprehensive analysis of sequence signature and BLAST and CRISPRTarget score, the 708 spacers were classified into three subtypes: Type I, Type II and Type III. Of them, Type I spacer referred to those linked with one gene segment, Type II spacer linked with two or more different gene segments, and Type III spacer undefined. This study examined the diversity of CRISPR/cas system in Shigella strains, demonstrated the main features of CRISPR structure and spacer classification, which provided critical information for elucidation of the mechanisms of spacer formation and exploration of the role the spacers play in the function of the CRISPR/cas system.

Immunologic Control of Diarrheal Disease Due to Enterotoxigenic Escherichia coli: Reactogenicity, Immunogenicity, and Efficacy Studies of Pili Vaccines

Science.gov (United States)

1981-09-01

bacterial hemagglutination teLhnique for detection of Shigella antibodies. J. Bacteriol. 91:463, 1966. -25- Appendix A 20. Levine, M.M. Legai and ethical...Gotschlich. E.C. Type 1 Escherichia coli pill: characterization of binding to monkey kidney cells. J. Exp. Med. 146:1132, 1977. 40. Santini, R., Jr., Sheehy

Free-Ranging Synanthropic Birds (Ardea alba and Columba livia domestica) as Carriers of Salmonella spp. and Diarrheagenic Escherichia coli in the Vicinity of an Urban Zoo.

Science.gov (United States)

de Oliveira, Mirela C V; Camargo, Beatriz Q; Cunha, Marcos P V; Saidenberg, Andre Becker; Teixeira, Rodrigo H F; Matajira, Carlos E C; Moreno, Luisa Z; Gomes, Vasco T M; Christ, Ana P G; Barbosa, Mikaela R F; Sato, Maria I Z; Moreno, Andrea M; Knöbl, Terezinha

2018-01-01

The presence of free-ranging urban birds is a risk factor for transmitting pathogens to captive animals and humans alike, including Salmonella spp. and diarrheagenic Escherichia coli. Cloacal swabs from 156 synanthropic Great egrets (Ardea alba) and feral pigeons (Columba livia domestica) that inhabit the surroundings of an urban zoo were processed for the identification of Salmonella spp. and diarrheagenic E. coli pathotypes. Bacterial species identification and genotypic characterization employed the matrix-assisted laser desorption ionization-time of flight mass spectrometry and PCR techniques, respectively, comparing their phylogenetic profiles through amplified fragment length polymorphism (AFLP) analysis. A total of 11 birds were positive for Salmonella Typhimurium (7%) and 9 individuals (5.8%) for diarrheagenic E. coli (enteropathogenic E. coli/Shiga-toxin producing E. coli [EPEC/STEC]) strains. S. Typhimurium strains presented highly similar AFLP profiles (85-100%), whereas EPEC/STEC strains showed more polymorphism. The results show free-ranging birds as carriers for both microorganisms in a zoo environment in Brazil for the first time and suggest these species as possible sources of infection to other animals as well as exposing personnel and visitors to potential zoonotic microorganisms. The presence of carriers highlights the importance of a surveillance system and the need for preventive measures to avoid attracting growing number of synanthropic avian species.

Characterization of the hemA-prs region of the Escherichia coli and Salmonella typhimurium chromosomes

DEFF Research Database (Denmark)

Post, David A.; Hove-Jensen, Bjarne; Switzer, Robert L.

1993-01-01

The prs gene, encoding phosphoribosylpyrophosphate synthetase, is preceded by a leader, which is 302 bp long in Escherichia coli and 417 bp in Salmonella typhimurium. A potential open reading frame (ORF) extends across the prs promoter and into the leader. The region between the prs coding region...... two promoters, the first promoter (P1) originating upstream of ORF 1, and expressing the prs gene in a tricistronic operon and a second promoter (P2), located within the ORF 2 coding frame, which transcribes the prs gene only. The transcripts encoding prs only were 20 times as abundant...... in the amount of message originating from the promoter P2....

Ethiop. J. Sci. & Technol. 8(1) 27-35, 2015 27 Prevalence and ...

African Journals Online (AJOL)

ABSTRACT. Diarrheal diseases remain the major cause of morbidity and mortality in children under five years of age. Salmonella and Shigella species are among the leading causes of diarrhea in children. The aim of this study was to determine the prevalence and antimicrobial profiles of Salmonella and Shigella spp. in ...

A sandwich-type optical immunosensor based on the alkaline phosphatase enzyme for Salmonella thypimurium detection

Science.gov (United States)

Widyastuti, E.; Puspitasari Schonherr, M. F.; Masruroh, A.; Anggraeni, R. A.; Nisak, Y. K.; Mursidah, S.

2018-03-01

Salmonella is pathogenic bacteria that caused foodborne diseases which being called Salmonellosis. Prevalence of Salmonellosis that being caused by Salmonella thypimurium in Indonesia is quite high. However, detection of Salmonella bacteria in food still limited, complicated, and required a lot time. Sensitive optical assay for Salmonella thypimurium paper based detection has been developed by integrating sandwich assay between antibody-antigen complex and alkaline phosphatase enzyme that produce visible bluish-purple colour with presence of NBT-BCIP substrate. The results showed that Limit of Quantitation of detection is 105 CFU mL-1 with detection time 15 minutes. Linearity test between Colour intensity that produced from Salmonella concentration presence on samples showed that detection has good linearity. Selectivity test exhibited excellent sensitivity with good discrimination against Escherichia coli.

Shigella isolates from the global enteric multicenter study inform vaccine development.

Science.gov (United States)

Livio, Sofie; Strockbine, Nancy A; Panchalingam, Sandra; Tennant, Sharon M; Barry, Eileen M; Marohn, Mark E; Antonio, Martin; Hossain, Anowar; Mandomando, Inacio; Ochieng, John B; Oundo, Joseph O; Qureshi, Shahida; Ramamurthy, Thandavarayan; Tamboura, Boubou; Adegbola, Richard A; Hossain, Mohammed Jahangir; Saha, Debasish; Sen, Sunil; Faruque, Abu Syed Golam; Alonso, Pedro L; Breiman, Robert F; Zaidi, Anita K M; Sur, Dipika; Sow, Samba O; Berkeley, Lynette Y; O'Reilly, Ciara E; Mintz, Eric D; Biswas, Kousick; Cohen, Dani; Farag, Tamer H; Nasrin, Dilruba; Wu, Yukun; Blackwelder, William C; Kotloff, Karen L; Nataro, James P; Levine, Myron M

2014-10-01

Shigella, a major diarrheal disease pathogen worldwide, is the target of vaccine development. The Global Enteric Multicenter Study (GEMS) investigated burden and etiology of moderate-to-severe diarrheal disease in children aged Shigella was 1 of the 4 most common pathogens across sites and age strata. GEMS Shigella serotypes are reviewed to guide vaccine development. Subjects' stool specimens/rectal swabs were transported to site laboratories in transport media and plated onto xylose lysine desoxycholate and MacConkey agar. Suspect Shigella colonies were identified by biochemical tests and agglutination with antisera. Shigella isolates were shipped to the GEMS Reference Laboratory (Baltimore, MD) for confirmation and serotyping of S. flexneri; one-third of isolates were sent to the Centers for Disease Control and Prevention for quality control. Shigella dysenteriae and S. boydii accounted for 5.0% and 5.4%, respectively, of 1130 Shigella case isolates; S. flexneri comprised 65.9% and S. sonnei 23.7%. Five serotypes/subserotypes comprised 89.4% of S. flexneri, including S. flexneri 2a, S. flexneri 6, S. flexneri 3a, S. flexneri 2b, and S. flexneri 1b. A broad-spectrum Shigella vaccine must protect against S. sonnei and 15 S. flexneri serotypes/subserotypes. A quadrivalent vaccine with O antigens from S. sonnei, S. flexneri 2a, S. flexneri 3a, and S. flexneri 6 can provide broad direct coverage against these most common serotypes and indirect coverage against all but 1 (rare) remaining subserotype through shared S. flexneri group antigens. © The Author 2014. Published by Oxford University Press on behalf of the Infectious Diseases Society of America.

Campylobacter, Salmonella, and Escherichia coli on broiler carcasses subjected to a high pH scald and low pH postpick chlorine dip.

Science.gov (United States)

Berrang, M E; Windham, W R; Meinersmann, R J

2011-04-01

The objective of this study was to determine the individual and combined effects of a high pH scald and a postpick chlorine dip on bacteria present on broiler carcasses. In each of 3 replications, a flock was sampled at several sites within a commercial broiler processing plant. Carcasses were sampled by whole carcass rinse before and after treated scalding at mean pH 9.89 or control scalding at mean pH 6.88. Other carcasses from the same flock run on both the treated and control scald lines were collected and sampled before and after a chlorine dip tank operated at mean total chlorine level of 83.3 mg/kg and pH 6.04. Rinses were cultured for numbers of Campylobacter and Escherichia coli and presence or absence of Salmonella. High pH scald was more effective than standard scald to lessen the prevalence and numbers of Campylobacter on broiler carcasses; a lower prevalence was maintained through the postpick chlorine dip tank. The pH of the scald tank made no difference in numbers of E. coli recovered from broiler carcasses at any tested point on the processing line. High pH scald was not more effective than standard scald to lessen Salmonella prevalence. Furthermore, it is unclear why the postpick chlorine dip effectively lessened Salmonella prevalence on only the control scald line. Although no evidence exists that these treatments have an additive effect when used in series, each treatment shows some promise individually. Further optimization may result in more effective decontamination of broiler carcasses.

Prevalence of Arcobacter, Escherichia coli, Staphylococcus aureus ...

African Journals Online (AJOL)

In this study, varying level of resistance of Escherichia coli 66(84.6%), Salmonella 6(100%) and Arcobacter 57(100%) to amoxicillin was observed. The susceptibility pattern indicates that the bacterial isolates exhibited a varying level of resistance to two or more antimicrobial agents with maximum resistance to amoxicillin.

Gene decay in Shigella as an incipient stage of host-adaptation.

Directory of Open Access Journals (Sweden)

Ye Feng

Full Text Available BACKGROUND: Many facultative bacterial pathogens have undergone extensive gene decay processes, possibly due to lack of selection pressure during evolutionary conversion from free-living to intracellular lifestyle. Shigella, the causative agents of human shigellosis, have arisen from different E. coli-like ancestors independently by convergent paths. As these bacteria all have lost large numbers of genes by mutation or deletion, they can be used as ideal models for systematically studying the process of gene function loss in different bacteria living under similar selection pressures. METHODOLOGIES/PRINCIPAL FINDINGS: We compared the sequenced Shigella genomes and re-defined decayed genes (pseudogenes plus deleted genes in these bacteria. Altogether, 85 genes are commonly decayed in the five analyzed Shigella strains and 1456 genes are decayed in at least one Shigella strain. Genes coding for carbon utilization, cell motility, transporter or membrane proteins are prone to be inactivated. Decayed genes tend to concentrate in certain operons rather than distribute averagely across the whole genome. Genes in the decayed operon accumulated more non-synonymous mutations than the rest genes and meanwhile have lower expression levels. CONCLUSIONS: Different Shigella lineages underwent convergent gene decay processes, and inactivation of one gene would lead to a lesser selection pressure for the other genes in the same operon. The pool of superfluous genes for Shigella may contain at least two thousand genes and the gene decay processes may still continue in Shigella until a minimum genome harboring only essential genes is reached.

Primary structure and mapping of the hupA gene of Salmonella typhimurium.

OpenAIRE

Higgins, N P; Hillyard, D

1988-01-01

In bacteria, the complex nucleoid structure is folded and maintained by negative superhelical tension and a set of type II DNA-binding proteins, also called histonelike proteins. The most abundant type II DNA-binding protein is HU. Southern blot analysis showed that Salmonella typhimurium contained two HU genes that corresponded to Escherichia coli genes hupA (encoding HU-2 protein) and hupB (encoding HU-1). Salmonella hupA was cloned, and the nucleotide sequence of the gene was determined. C...

Cladophora (Chlorophyta) spp. Harbor Human Bacterial Pathogens in Nearshore Water of Lake Michigan†

Science.gov (United States)

Ishii, Satoshi; Yan, Tao; Shively, Dawn A.; Byappanahalli, Muruleedhara N.; Whitman, Richard L.; Sadowsky, Michael J.

2006-01-01

Cladophora glomerata, a macrophytic green alga, is commonly found in the Great Lakes, and significant accumulations occur along shorelines during the summer months. Recently, Cladophora has been shown to harbor high densities of the fecal indicator bacteria Escherichia coli and enterococci. Cladophora may also harbor human pathogens; however, until now, no studies to address this question have been performed. In the present study, we determined whether attached Cladophora, obtained from the Lake Michigan and Burns Ditch (Little Calumet River, Indiana) sides of a breakwater during the summers of 2004 and 2005, harbored the bacterial pathogens Shiga toxin-producing Escherichia coli (STEC), Salmonella, Shigella, and Campylobacter. The presence of potential pathogens and numbers of organisms were determined by using cultural methods and by using conventional PCR, most-probable-number PCR (MPN-PCR), and quantitative PCR (QPCR) performed with genus- and toxin-specific primers and probes. While Shigella and STEC were detected in 100% and 25%, respectively, of the algal samples obtained near Burns Ditch in 2004, the same pathogens were not detected in samples collected in 2005. MPN-PCR and QPCR allowed enumeration of Salmonella in 40 to 80% of the ditch- and lakeside samples, respectively, and the densities were up to 1.6 × 103 cells per g Cladophora. Similarly, these PCR methods allowed enumeration of up to 5.4 × 102 Campylobacter cells/g Cladophora in 60 to 100% of lake- and ditchside samples. The Campylobacter densities were significantly higher (P Cladophora samples than in the ditchside Cladophora samples. DNA fingerprint analyses indicated that genotypically identical Salmonella isolates were associated with geographically and temporally distinct Cladophora samples. However, Campylobacter isolates were genetically diverse. Since animal hosts are thought to be the primary habitat for Campylobacter and Salmonella species, our results suggest that Cladophora is a

Cladophora (Chlorophyta) spp. harbor human bacterial pathogens in nearshore water of Lake Michigan.

Science.gov (United States)

Ishii, Satoshi; Yan, Tao; Shively, Dawn A; Byappanahalli, Muruleedhara N; Whitman, Richard L; Sadowsky, Michael J

2006-07-01

Cladophora glomerata, a macrophytic green alga, is commonly found in the Great Lakes, and significant accumulations occur along shorelines during the summer months. Recently, Cladophora has been shown to harbor high densities of the fecal indicator bacteria Escherichia coli and enterococci. Cladophora may also harbor human pathogens; however, until now, no studies to address this question have been performed. In the present study, we determined whether attached Cladophora, obtained from the Lake Michigan and Burns Ditch (Little Calumet River, Indiana) sides of a breakwater during the summers of 2004 and 2005, harbored the bacterial pathogens Shiga toxin-producing Escherichia coli (STEC), Salmonella, Shigella, and Campylobacter. The presence of potential pathogens and numbers of organisms were determined by using cultural methods and by using conventional PCR, most-probable-number PCR (MPN-PCR), and quantitative PCR (QPCR) performed with genus- and toxin-specific primers and probes. While Shigella and STEC were detected in 100% and 25%, respectively, of the algal samples obtained near Burns Ditch in 2004, the same pathogens were not detected in samples collected in 2005. MPN-PCR and QPCR allowed enumeration of Salmonella in 40 to 80% of the ditch- and lakeside samples, respectively, and the densities were up to 1.6 x 10(3) cells per g Cladophora. Similarly, these PCR methods allowed enumeration of up to 5.4 x 10(2) Campylobacter cells/g Cladophora in 60 to 100% of lake- and ditchside samples. The Campylobacter densities were significantly higher (P Cladophora samples than in the ditchside Cladophora samples. DNA fingerprint analyses indicated that genotypically identical Salmonella isolates were associated with geographically and temporally distinct Cladophora samples. However, Campylobacter isolates were genetically diverse. Since animal hosts are thought to be the primary habitat for Campylobacter and Salmonella species, our results suggest that Cladophora is a

Intradermal delivery of Shigella IpaB and IpaD type III secretion proteins: kinetics of cell recruitment and antigen uptake, mucosal and systemic immunity, and protection across serotypes.

Science.gov (United States)

Heine, Shannon J; Diaz-McNair, Jovita; Andar, Abhay U; Drachenberg, Cinthia B; van de Verg, Lillian; Walker, Richard; Picking, Wendy L; Pasetti, Marcela F

2014-02-15

Shigella is one of the leading pathogens contributing to the vast pediatric diarrheal disease burden in low-income countries. No licensed vaccine is available, and the existing candidates are only partially effective and serotype specific. Shigella type III secretion system proteins IpaB and IpaD, which are conserved across Shigella spp., are candidates for a broadly protective, subunit-based vaccine. In this study, we investigated the immunogenicity and protective efficacy of IpaB and IpaD administered intradermally (i.d.) with a double-mutant of the Escherichia coli heat-labile enterotoxin (dmLT) adjuvant using microneedles. Different dosage levels of IpaB and IpaD, with or without dmLT, were tested in mice. Vaccine delivery into the dermis, recruitment of neutrophils, macrophages, dendritic cells, and Langerhans cells, and colocalization of vaccine Ag within skin-activated APC were demonstrated through histology and immunofluorescence microscopy. Ag-loaded neutrophils, macrophages, dendritic cells, and Langerhans cells remained in the tissue at least 1 wk. IpaB, IpaD, and dmLT-specific serum IgG- and IgG-secreting cells were produced following i.d. immunization. The protective efficacy was 70% against Shigella flexneri and 50% against Shigella sonnei. Similar results were obtained when the vaccine was administered intranasally, with the i.d. route requiring 25-40 times lower doses. Distinctively, IgG was detected in mucosal secretions; secretory IgA, as well as mucosal and systemic IgA Ab-secreting cells, were seemingly absent. Vaccine-induced T cells produced IFN-γ, IL-2, TNF-α, IL-17, IL-4, IL-5, and IL-10. These results demonstrate the potential of i.d. vaccination with IpaB and IpaD to prevent Shigella infection and support further studies in humans.

Pathogenesis of Escherichia coli O157:H7 Strain 86-24 Following Oral Infection of BALB/c Mice with an Intact Commensal Flora

Science.gov (United States)

2010-01-01

inocula of these organisms are also needed to achieve colonization and disease in monkeys [29] and rabbits [30]. Our data suggested that the predominate...SB. Escherichia coli 0157:H7 srrains associared wirh haemorrhagic colitis in rhe Unired Srares produce a Shigella dysenreriae 1 (SHIGA) like...611-20. (29) Kang G, Pulimood AB, Koshi R. Hull A, Acheson D. Rajan P. er al. A monkey model for enrerohemorrhagic Escherichia coli infection. j

Temperature Regulation of Shigella Virulence: Identification of Temperature-Regulated Shigella Invasion Genes by the Isolation of inv::lacZ Operon Fusions and the Characterization of the Virulence Gene Regulator virR

Science.gov (United States)

1991-04-10

conserved in virulent strains of Shigella and EIEC. Evaluation of the serum Immune response to Shigella proteins in Rhesus monkeys and humans revealed...sera from both humans and monkeys following a Shigella infection (Hale et al. , 1985; Oaks et al., 1986). Moreover, Tn5 insertions In various...Microbiology, Washington, D.C. 108. Oaks, E, V,, T, L. Hale, and S. B. Formal. 1986. Serum Immune response to Shigella protein antigens In Rhesus monkeys

Effect of guinea pig or monkey colonic mucus on Shigella aggregation and invasion of HeLa cells by Shigella flexneri 1b and 2a.

OpenAIRE

Dinari, G; Hale, T L; Washington, O; Formal, S B

1986-01-01

The effects of guinea pig and rhesus monkey colonic mucus preparations on Shigella aggregation and invasion of HeLa cell monolayers by Shigella flexneri serotype 1b, 2a, and 5 strains were investigated. Guinea pig mucus caused agglutination of S. flexneri serotype 1b but not of S. flexneri serotype 2a or 5. Guinea pig mucus also inhibited HeLa cell invasion by S. flexneri serotypes 1b and 2a. Monkey mucus neither agglutinated any Shigella strain nor inhibited HeLa cell invasion.

Update on: Shigella new serogroups/serotypes and their antimicrobial resistance.

Science.gov (United States)

Muthuirulandi Sethuvel, D P; Devanga Ragupathi, N K; Anandan, S; Veeraraghavan, B

2017-01-01

Shigellosis represents a major burden of disease in developing countries. A low infectious dose allows the disease to be spread effectively. Although shigellosis is mostly a self-limiting disease, antibiotics are recommended to reduce deaths, disease symptoms and organism-shedding time. However, in India, antimicrobial resistance among the genus Shigella is more common than among any other enteric bacteria. Notably, new serotypes or subserotypes in Shigella are reported from various parts of the world. Identification of new subserotypes of Shigella spp. is becoming a major issue as these strains are nontypeable by conventional serotyping. The commercially available antisera may not cover all possible epitopes of the O lipopolysaccharide antigen of Shigella serotypes. Therefore, molecular methods which most closely approach the resolution of full serotyping are necessary to identify such strains. In addition, the knowledge of a prevalent serotype in various geographic regions may assist in formulating strategies such as the development of a vaccine to prevent infection especially when the immunity to disease is serotype specific, and to understand the disease burden caused by new Shigella serotypes. © 2016 The Society for Applied Microbiology.

Applying Mathematical Tools to Accelerate Vaccine Development: Modeling Shigella Immune Dynamics

Science.gov (United States)

Davis, Courtney L.; Wahid, Rezwanul; Toapanta, Franklin R.; Simon, Jakub K.

2013-01-01

We establish a mathematical framework for studying immune interactions with Shigella, a bacteria that kills over one million people worldwide every year. The long-term goal of this novel approach is to inform Shigella vaccine design by elucidating which immune components and bacterial targets are crucial for establishing Shigella immunity. Our delay differential equation model focuses on antibody and B cell responses directed against antigens like lipopolysaccharide in Shigella’s outer membrane. We find that antibody-based vaccines targeting only surface antigens cannot elicit sufficient immunity for protection. Additional boosting prior to infection would require a four-orders-of-magnitude increase in antibodies to sufficiently prevent epithelial invasion. However, boosting anti-LPS B memory can confer protection, which suggests these cells may correlate with immunity. We see that IgA antibodies are slightly more effective per molecule than IgG, but more total IgA is required due to spatial functionality. An extension of the model reveals that targeting both LPS and epithelial entry proteins is a promising avenue to advance vaccine development. This paper underscores the importance of multifaceted immune targeting in creating an effective Shigella vaccine. It introduces mathematical models to the Shigella vaccine development effort and lays a foundation for joint theoretical/experimental/clinical approaches to Shigella vaccine design. PMID:23589755

How Do the Virulence Factors of Shigella Work Together to Cause Disease?

Science.gov (United States)

Mattock, Emily; Blocker, Ariel J

2017-01-01

Shigella is the major cause of bacillary dysentery world-wide. It is divided into four species, named S. flexneri, S. sonnei, S. dysenteriae , and S. boydii , which are distinct genomically and in their ability to cause disease. Shigellosis, the clinical presentation of Shigella infection, is characterized by watery diarrhea, abdominal cramps, and fever. Shigella 's ability to cause disease has been attributed to virulence factors, which are encoded on chromosomal pathogenicity islands and the virulence plasmid. However, information on these virulence factors is not often brought together to create a detailed picture of infection, and how this translates into shigellosis symptoms. Firstly, Shigella secretes virulence factors that induce severe inflammation and mediate enterotoxic effects on the colon, producing the classic watery diarrhea seen early in infection. Secondly, Shigella injects virulence effectors into epithelial cells via its Type III Secretion System to subvert the host cell structure and function. This allows invasion of epithelial cells, establishing a replicative niche, and causes erratic destruction of the colonic epithelium. Thirdly, Shigella produces effectors to down-regulate inflammation and the innate immune response. This promotes infection and limits the adaptive immune response, causing the host to remain partially susceptible to re-infection. Combinations of these virulence factors may contribute to the different symptoms and infection capabilities of the diverse Shigella species, in addition to distinct transmission patterns. Further investigation of the dominant species causing disease, using whole-genome sequencing and genotyping, will allow comparison and identification of crucial virulence factors and may contribute to the production of a pan- Shigella vaccine.

Shigella Iron Acquisition Systems and their Regulation.

Science.gov (United States)

Wei, Yahan; Murphy, Erin R

2016-01-01

Survival of Shigella within the host is strictly dependent on the ability of the pathogen to acquire essential nutrients, such as iron. As an innate immune defense against invading pathogens, the level of bio-available iron within the human host is maintained at exceeding low levels, by sequestration of the element within heme and other host iron-binding compounds. In response to sequestration mediated iron limitation, Shigella produce multiple iron-uptake systems that each function to facilitate the utilization of a specific host-associated source of nutrient iron. As a mechanism to balance the essential need for iron and the toxicity of the element when in excess, the production of bacterial iron acquisition systems is tightly regulated by a variety of molecular mechanisms. This review summarizes the current state of knowledge on the iron-uptake systems produced by Shigella species, their distribution within the genus, and the molecular mechanisms that regulate their production.

Evaluation of immunogenicity and protective efficacy of orally delivered Shigella type III secretion system proteins IpaB and IpaD.

Science.gov (United States)

Heine, Shannon J; Diaz-McNair, Jovita; Martinez-Becerra, Francisco J; Choudhari, Shyamal P; Clements, John D; Picking, Wendy L; Pasetti, Marcela F

2013-06-19

Shigella spp. are food- and water-borne pathogens that cause shigellosis, a severe diarrheal and dysenteric disease that is associated with a high morbidity and mortality in resource-poor countries. No licensed vaccine is available to prevent shigellosis. We have recently demonstrated that Shigella invasion plasmid antigens (Ipas), IpaB and IpaD, which are components of the bacterial type III secretion system (TTSS), can prevent infection in a mouse model of intranasal immunization and lethal pulmonary challenge. Because they are conserved across Shigella spp. and highly immunogenic, these proteins are excellent candidates for a cross-protective vaccine. Ideally, such a vaccine could be administered to humans orally to induce mucosal and systemic immunity. In this study, we investigated the immunogenicity and protective efficacy of Shigella IpaB and IpaD administered orally with a double mutant of the Escherichia coli heat labile toxin (dmLT) as a mucosal adjuvant. We characterized the immune responses induced by oral vs. intranasal immunization and the protective efficacy using a mouse pulmonary infection model. Serum IgG and fecal IgA against IpaB were induced after oral immunization. These responses, however, were lower than those obtained after intranasal immunization despite a 100-fold dosage increase. The level of protection induced by oral immunization with IpaB and IpaD was 40%, while intranasal immunization resulted in 90% protective efficacy. IpaB- and IpaD-specific IgA antibody-secreting cells in the lungs and spleen and T-cell-derived IL-2, IL-5, IL-17 and IL-10 were associated with protection. These results demonstrate the immunogenicity of orally administered IpaB and IpaD and support further studies in humans. Copyright © 2013 Elsevier Ltd. All rights reserved.

Clostridium difficile Recombinant Toxin A Repeating Units as a Carrier Protein for Conjugate Vaccines: Studies of Pneumococcal Type 14, Escherichia coli K1, and Shigella flexneri Type 2a Polysaccharides in Mice

Science.gov (United States)

Pavliakova, Danka; Moncrief, J. Scott; Lyerly, David M.; Schiffman, Gerald; Bryla, Dolores A.; Robbins, John B.; Schneerson, Rachel

2000-01-01

Unlike the native protein, a nontoxic peptide (repeating unit of the native toxin designated rARU) from Clostridium difficile toxin A (CDTA) afforded an antigen that could be bound covalently to the surface polysaccharides of pneumococcus type 14, Shigella flexneri type 2a, and Escherichia coli K1. The yields of these polysaccharide-protein conjugates were significantly increased by prior treatment of rARU with succinic anhydride. Conjugates, prepared with rARU or succinylated (rARUsucc), were administered to mice by a clinically relevant dosage and immunization scheme. All conjugates elicited high levels of serum immunoglobulin G both to the polysaccharides and to CDTA. Conjugate-induced anti-CDTA had neutralizing activity in vitro and protected mice challenged with CDTA, similar to the rARU alone. Conjugates prepared with succinylated rARU, therefore, have potential for serving both as effective carrier proteins for polysaccharides and for preventing enteric disease caused by C. difficile. PMID:10722615

Customizable PCR-microplate array for differential identification of multiple pathogens.

Science.gov (United States)

Woubit, Abdela; Yehualaeshet, Teshome; Roberts, Sherrelle; Graham, Martha; Kim, Moonil; Samuel, Temesgen

2013-11-01

Customizable PCR-microplate arrays were developed for the rapid identification of Salmonella Typhimurium, Salmonella Saintpaul, Salmonella Typhi, Shigella dysenteriae, Escherichia coli O157:H7, Francisella tularensis subsp. tularensis, Francisella tularensis subsp. novicida, Vibrio cholerae, Vibrio parahaemolyticus, Yersinia pestis, and Yersinia pseudotuberculosis. Previously, we identified highly specific primers targeting each of these pathogens. Here, we report the development of customizable PCR-microplate arrays for simultaneous identification of the pathogens using the primers identified. A mixed aliquot of genomic DNA from 38 strains was used to validate three PCR-microplate array formats. Identical PCR conditions were used to run all the samples on the three formats. Specific amplifications were obtained on all three custom plates. In preliminary tests performed to evaluate the sensitivity of these assays in samples inoculated in the laboratory with Salmonella Typhimurium, amplifications were obtained from 1 g of beef hot dog inoculated at as low as 9 CFU/ml or from milk inoculated at as low as 78 CFU/ml. Such microplate arrays could be valuable tools for initial identification or secondary confirmation of contamination by these pathogens.

Antimicrobial resistence of Shigella species isolated during 2004 and 2005 from selected sites in Zimbabwe.

Science.gov (United States)

Ndlovu, N; Tarupiwa, A; Mudzori, J T

2006-01-01

To determine the predominant serotype and antibiotic sensitivity pattern of Shigella isolates during 2004 and 2005 in Zimbabwe. Cross sectional study. National Microbiology Reference Laboratory (NMRL), Harare, Zimbabwe. 259 clinical isolates of Shigella species isolated during 2004 and 2005 in Zimbabwe were studied. These samples had been referred to the NMRL for further testing. Serotype and antibiotic sensitivity pattern of Shigella species. Of the 259 clinical isolates of Shigella tested the following species were serotyped; 141 (54.4%) were S. flexneri; 70 (27%) S. sonnei; 38 (14.7%) S. dysenteriae and 10 (3.9%) S. boydii. About 4% of all Shigella isolates tested showed full sensitivity to commonly used antibiotics, 20.8% were resistant to one antibiotic only while 75.3% were resistant to at least two antibiotics. The most common resistance among Shigella species was to cotrimoxazole (89%), tetracycline (73%), ampicillin (49%) and chloramphenicol (41%). High susceptibility among Shigella species was observed to nalidixic acid (86%), ciprofloxacin (99%) and ceftazidine (99%). There was a low drug resistance of Shigella species to nalidixic acid, a drug of choice in Zimbabwe, except among Shigella dysenteriae type 1 strains. Continuous monitoring of the susceptibility patterns of Shigella species is important in order to detect the emergence of drug resistance and to update guidelines for antibiotic treatment in shigellosis.

[Molecular characteristics of Clustered Regularly Interspaced Short Palindromic Repeat in Shigella].

Science.gov (United States)

Xue, Zerun; Wang, Yingfang; Duan, Guangcai; Yang, Haiyan; Xi, Yuanlin; Wang, Pengfei; Wang, Linlin; Guo, Xiangjiao

2015-08-01

To detect the molecular characteristics of Clustered Regularly Interspaced Short Palindromic Repeat (CRISPR) in Shigella and to analyze the distribution of CRISPR related to the time of isolation. Of the 52 Shigella strains, 41 were isolated from Henan, 6 from Jiangxi and 5 isolated from Beijing. Both CRISPR locus of S1, S2, S3 and S4 in Shigella were detected by polymerase chain reaction (PCR). The PCR products were sequenced and compared. The positive rates of CRISPR locus in Shigella were 33.69% (S1), 50.00% (S2), 82.69% (S3) and 73.08% (S4), respectively. Two subtypes were discovered in S1 and S3 locus. Three subtypes were discovered in S2 locus. Four different subtypes were discovered in S4 locus. The isolates from Henan strains were divided into two groups by the time of isolation. Distributions of S1 were different, before or after 2004, on Shigella. S1 could not be detected after 2004. There were no statistical differences of S2, S3 and S4 in two groups. Different CRISPR subtypes or Shigella were discovered. A significant correlation was noticed between the CRISPR S1 related to the time of isolation but not between S2, S3 or S4 on the time of isolation.

Assessment of Salmonella spp. and Escherichia coli O157:H7 growth on lettuce exposed to isothermal and non-isothermal conditions.

Science.gov (United States)

de Oliveira Elias, Susana; Noronha, Tiago Baptista; Tondo, Eduardo Cesar

2018-06-01

This study aimed to assess the growth of Salmonella and Escherichia coli O157:H7 on lettuce exposed to isothermal and non-isothermal conditions. Pathogens were inoculated on lettuce separately and stored under isothermal condition at 5 °C, 10 °C, 25 °C, 37 °C for both bacteria, at 40 °C for Salmonella and 42 °C for E. coli O157:H7. Growth curves were built by fitting the data to the Baranyi's DMFit, generating R 2 values greater than 0.92 for primary models. Secondary models were fitted with Ratkowsky equations, generating R 2 values higher than 0.91 and RMSE lower than 0.1. Experimental data showed that both bacteria could grow at all temperatures. Also, the growth of both pathogens under non-isothermal conditions was studied simulating temperatures found from harvest to supermarkets in Brazil. Models were analysed by R 2 , RMSE, bias factor (Bf) and accuracy factor (Af). Salmonella and E. coli O157:H7 were able to grow in this temperature profile and the models could predict the behavior of these microorganisms on lettuce under isothermal and non-isothermal conditions. Based on the results, a negligible growth time (ς) was proposed to provide the time which lettuce could be exposed to a specific temperature and do not present an expressive growth of bacteria. The ς was developed based on Baranyi's primary model equation and on growth potential concept. ς is the value of lag phase added of the time necessary to population grow 0.5 log CFU/g. The ς of lettuce exposed to 37 °C was 1.3 h, while at 5 °C was 3.3 days. Copyright © 2017 Elsevier Ltd. All rights reserved.

Complete genome sequence of Shigella flexneri 5b and comparison with Shigella flexneri 2a

Directory of Open Access Journals (Sweden)

Xue Ying

2006-07-01

Full Text Available Abstract Background Shigella bacteria cause dysentery, which remains a significant threat to public health. Shigella flexneri is the most common species in both developing and developed countries. Five Shigella genomes have been sequenced, revealing dynamic and diverse features. To investigate the intra-species diversity of S. flexneri genomes further, we have sequenced the complete genome of S. flexneri 5b strain 8401 (abbreviated Sf8401 and compared it with S. flexneri 2a (Sf301. Results The Sf8401 chromosome is 4.5-Mb in size, a little smaller than that of Sf301, mainly because the former lacks the SHI-1 pathogenicity island (PAI. Compared with Sf301, there are 6 inversions and one translocation in Sf8401, which are probably mediated by insertion sequences (IS. There are clear differences in the known PAIs between these two genomes. The bacteriophage SfV segment remaining in SHI-O of Sf8401 is clearly larger than the remnants of bacteriophage SfII in Sf301. SHI-1 is absent from Sf8401 but a specific related protein is found next to the pheV locus. SHI-2 is involved in one intra-replichore inversion near the origin of replication, which may change the expression of iut/iuc genes. Moreover, genes related to the glycine-betaine biosynthesis pathway are present only in Sf8401 among the known Shigella genomes. Conclusion Our data show that the two S. flexneri genomes are very similar, which suggests a high level of structural and functional conservation between the two serotypes. The differences reflect different selection pressures during evolution. The ancestor of S. flexneri probably acquired SHI-1 and SHI-2 before SHI-O was integrated and the serotypes diverged. SHI-1 was subsequently deleted from the S. flexneri 5b genome by recombination, but stabilized in the S. flexneri 2a genome. These events may have contributed to the differences in pathogenicity and epidemicity between the two serotypes of S. flexneri.

Export of the Virulence Factors from Shigella Flexneri and Characterization of the mxi loci

Science.gov (United States)

1992-07-20

studied in a monkey model for Shigella infection. The in vitro assays just described have been routinely used to closely examine each of the steps in...to Shigella protein antigens in Rhesus monkeys and humans infected with Shigella spp. Infect. Immun_ 53:57-63. Oaks, E. V. , and K. R. Turbyfill...thesis manuscript ent itled: ~Export of Virulence Faclors from Shigella f/exneri and characterization of the mxi loci. ~ beyond brief excerpts is

Effects of dietary additives (potassium diformate/organic acids) as well as influences of grinding intensity (coarse/fine) of diets for weaned piglets experimentally infected with Salmonella Derby or Escherichia coli.

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Taube, V A; Neu, M E; Hassan, Y; Verspohl, J; Beyerbach, M; Kamphues, J

2009-06-01

The aim of this study was to examine whether and to what extent the addition of potassium diformate (pdf) or free organic acids (fpa) to the diet and the grinding intensity might affect the course of infection and the passage of orally applied Salmonella and Escherichia coli in pigs. Experiments were carried out using 80 reared piglets allotted to four groups. Pigs were fed pelleted diets ad libitum (except during a 15 h feed-withholding-period before infection). The control diet contained finely ground cereals (2 mm screen). To two test diets (also finely ground) 1.2% pdf, 0.9% organic acids (75% formic and 25% propionic acid, fpa) respectively were added. The fourth diet (without acids) was based on coarsely ground cereals (6-mm screen). After experimental infection alternately with S. Derby or E. coli, the course of infection was examined (rectal swab technique). Pigs were sacrificed 4-5 h after a further oral application of approximately 10(9)-10(10) CFU S. Derby or E. coli to determine the counts of Salmonella or E. coli in chyme (classical culture methods). Adding pdf or fpa to the diet led to reduced Salmonella shedding and resulted in significantly lower counts of Salmonella and E. coli in the stomach content indicating an improved efficacy of the stomach barrier. In the distal parts of the digestive tract, the effect was less obvious concerning counts of E. coli, whereas counts of Salmonella were reduced markedly as well. The diet based on coarsely ground cereals failed to demonstrate positive effects concerning infection and passage of orally applied bacteria as well, but this diet was also pelleted and showed unintentionally, comparable amounts of fine particles. Results obtained in this study allow the recommendation of using pdf or organic acids as additives when dietary measures against Salmonella or E. coli in pigs are required.

A Review of Current Research and Knowledge Gaps in the Epidemiology of Shiga Toxin-Producing Escherichia coli and Salmonella spp. in Trinidad and Tobago.

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Persad, Anil K; LeJeune, Jeffrey

2018-04-17

Salmonella and Shiga toxin-producing Escherichia coli are two of the main causes of foodborne disease globally, and while they have been implicated as possible causes of foodborne disease within the Caribbean region, the actual incidence is unknown. Trinidad and Tobago, one of the larger countries in the Caribbean, has an estimated annual foodborne disease burden of over 100,000 cases and, similar to other countries, the etiology of most of these cases is unknown. Both pathogens can reside as part of the normal gastrointestinal microflora of many wild and domestic animals, with animals acting as reservoirs, spillover hosts, or dead-end hosts. Carriage in animal species can be asymptomatic or, in the case of Salmonella in particular, there may be clinical manifestation in animals, which resemble the disease seen in humans. In this review, we will focus on the epidemiology of these two foodborne pathogens in Trinidad and Tobago and identify any knowledge gaps in the published literature. The filling of this critical knowledge void is essential for the development and implementation of appropriate mechanisms to reduce the dissemination and transmission of these pathogens, not only in Trinidad and Tobago, but also in the wider Caribbean.

Genetic plasticity of the Shigella virulence plasmid is mediated by intra- and inter-molecular events between insertion sequences.

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Pilla, Giulia; McVicker, Gareth; Tang, Christoph M

2017-09-01

Acquisition of a single copy, large virulence plasmid, pINV, led to the emergence of Shigella spp. from Escherichia coli. The plasmid encodes a Type III secretion system (T3SS) on a 30 kb pathogenicity island (PAI), and is maintained in a bacterial population through a series of toxin:antitoxin (TA) systems which mediate post-segregational killing (PSK). The T3SS imposes a significant cost on the bacterium, and strains which have lost the plasmid and/or genes encoding the T3SS grow faster than wild-type strains in the laboratory, and fail to bind the indicator dye Congo Red (CR). Our aim was to define the molecular events in Shigella flexneri that cause loss of Type III secretion (T3S), and to examine whether TA systems exert positional effects on pINV. During growth at 37°C, we found that deletions of regions of the plasmid including the PAI lead to the emergence of CR-negative colonies; deletions occur through intra-molecular recombination events between insertion sequences (ISs) flanking the PAI. Furthermore, by repositioning MvpAT (which belongs to the VapBC family of TA systems) near the PAI, we demonstrate that the location of this TA system alters the rearrangements that lead to loss of T3S, indicating that MvpAT acts both globally (by reducing loss of pINV through PSK) as well as locally (by preventing loss of adjacent sequences). During growth at environmental temperatures, we show for the first time that pINV spontaneously integrates into different sites in the chromosome, and this is mediated by inter-molecular events involving IS1294. Integration leads to reduced PAI gene expression and impaired secretion through the T3SS, while excision of pINV from the chromosome restores T3SS function. Therefore, pINV integration provides a reversible mechanism for Shigella to circumvent the metabolic burden imposed by pINV. Intra- and inter-molecular events between ISs, which are abundant in Shigella spp., mediate plasticity of S. flexneri pINV.

Antimicrobial Effects of Psidium Guajava Extract as One Mechanism of its Antidiarrhoeal Action

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Lutterodt, G.D.; Ismail, A.; Basheer, R.H.; Baharudin, H. Mohd.

1999-01-01

A morphine-like spasmolytic action (not naloxone reversible; involving the inhibition of acetylcholine release) and also effects on the transmural transport of electrolytes (Na+ and K+) and water have been reported as possible modes of the antidiarrhoeal action of polar fractions of Psidium guajava leaf extractives. The objective for this study was to verify if the reported modes of the antidiarrhoeal action should be broadened to include direct antimicrobial actions on some of the more common bacteria known to cause toxin-induced acute diarrhoea. Serial dilutions of a water-soluble, freeze-dried methanolic extract were tested on 10 such organisms, grown separately on nutrient agar plates, to determine the minimum inhibitory concentration (MIC) for each of these bacteria. These included the causative agents for (i) enteric fever (Salmonella typhi, Salmonella paratyphi A, Salmonella paratyphi B and Salmonella paratyphi C), (ii) food poisoning (Salmonella typhimurium and Staphylococcus aureus), (iii) dysentery (Shigella dysenteriae, Shigella flexneri and Shigella sonnei), and (iv) cholera (Vibrio cholerae). The growth of all these organisms was inhibited at the MIC of 10mg/ml of the extract, which is equivalent to 2.5μg/ml of active extractable flavonoids. The most sensitive organisms (MIC = 1mg/ml) were Staphylococcus aureus, Vibrio cholerae and Shigella flexneri. PMID:22589684

Protective immunity by oral immunization with heat-killed Shigella strains in a guinea pig colitis model.

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Barman, Soumik; Koley, Hemanta; Ramamurthy, Thandavarayan; Chakrabarti, Manoj Kumar; Shinoda, Sumio; Nair, Gopinath Balakrish; Takeda, Yoshifumi

2013-11-01

The protective efficacy of and immune response to heat-killed cells of monovalent and hexavalent mixtures of six serogroups/serotypes of Shigella strains (Shigella dysenteriae 1, Shigella flexneri 2a, S. flexneri 3a, S. flexneri 6, Shigella boydii 4, and Shigella sonnei) were examined in a guinea pig colitis model. A monovalent or hexavalent mixture containing 1 × 10(7) of each serogroup/serotype of heat-killed Shigella cells was administered orally on Days 0, 7, 14 and 21. On Day 28, the immunized animals were challenged rectally with 1 × 10(9) live virulent cells of each of the six Shigella serogroups/serotypes. In all immunized groups, significant levels of protection were observed after these challenges. The serum titers of IgG and IgA against the lipopolysaccharide of each of the six Shigella serogroups/serotypes increased exponential during the course of immunization. High IgA titers against the lipopolysaccharide of each of the six Shigella serogroups/serotypes were also observed in intestinal lavage fluid from all immunized animals. These data indicate that a hexavalent mixture of heat-killed cells of the six Shigella serogroups/serotypes studied would be a possible broad-spectrum candidate vaccine against shigellosis. © 2013 The Societies and Wiley Publishing Asia Pty Ltd.

The infectious hypoxia: occurrence and causes during Shigella infection.

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Arena, Ellen T; Tinevez, Jean-Yves; Nigro, Giulia; Sansonetti, Philippe J; Marteyn, Benoit S

2017-03-01

Hypoxia is defined as a tissue oxygenation status below physiological needs. During Shigella infection, an infectious hypoxia is induced within foci of infection. In this review, we discuss how Shigella physiology and virulence are modulated and how the main recruited immune cells, the neutrophils, adapt to this environment. Copyright © 2016 Institut Pasteur. Published by Elsevier Masson SAS. All rights reserved.

Multifactor Regulation of the MdtJI Polyamine Transporter in Shigella.

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Adriano Leuzzi

Full Text Available The polyamine profile of Shigella, the etiological agent of bacillary dysentery in humans, differs markedly from that of E. coli, its innocuous commensal ancestor. Pathoadaptive mutations such as the loss of cadaverine and the increase of spermidine favour the full expression of the virulent phenotype of Shigella. Spermidine levels affect the expression of the MdtJI complex, a recently identified efflux pump belonging to the small multi-drug resistance family of transporters. In the present study, we have addressed the regulation of the mdtJI operon in Shigella by asking which factors influence its expression as compared to E. coli. In particular, after identifying the mdtJI promoter by primer extension analysis, in vivo transcription assays and gel-retardation experiments were carried out to get insight on the silencing of mdtJI in E. coli. The results indicate that H-NS, a major nucleoid protein, plays a key role in repressing the mdtJI operon by direct binding to the regulatory region. In the Shigella background mdtJI expression is increased by the high levels of spermidine typically found in this microorganism and by VirF, the plasmid-encoded regulator of the Shigella virulence regulatory cascade. We also show that the expression of mdtJI is stimulated by bile components. Functional analyses reveal that MdtJI is able to promote the excretion of putrescine, the spermidine precursor. This leads us to consider the MdtJI complex as a possible safety valve allowing Shigella to maintain spermidine to a level optimally suited to survival within infected macrophages and, at the same time, prevent toxicity due to spermidine over-accumulation.

Genetic characterization of Shigella spp. isolated from diarrhoeal and asymptomatic children.

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Ghosh, Santanu; Pazhani, Gururaja P; Niyogi, Swapan Kumar; Nataro, James P; Ramamurthy, Thandavarayan

2014-07-01

Phenotypic and genetic characteristics of Shigella spp. isolated from diarrhoeal and asymptomatic children aged up to 5 years were analysed in this study. In total, 91 and 17 isolates were identified from diarrhoeal (case) and asymptomatic (control) children, respectively. All the isolates were tested for antimicrobial resistance, the presence of integrons, plasmid-mediated quinolone resistance (PMQR), virulence-associated genes and Shigella pathogenicity island (SH-PAI). The majority of the Shigella spp. from cases (68.1%) and controls (82.3%) were found to be resistant to fluoroquinolones. Integron carriage was detected more in cases (76.9%) than in controls (35.5%). Atypical class 1 integron was detected exclusively in Shigella flexneri from cases but not from the controls. PMQR genes such as aac(6')-Ib-cr and qnrS1 were detected in 82.4 and 14.3% of the isolates from cases and in 53 and 17.6% in controls, respectively. Shigella isolates from cases as well as from controls were positive for the invasive plasmid antigen H-encoding gene ipaH. The other virulence genes such as virF, sat, setA, setB, sen and ial were detected in Shigella isolates in 80.2, 49.4, 27.4, 27.4, 80.2 and 79.1% of cases and in 64.7, 52.9, 17.6, 17.6, 64.7 and 64.7% of controls, respectively. The entire SH-PAI was detected in S. flexneri serotype 2a from cases and controls. In an isolate from a control child, the SH-PAI was truncated. Integrons, PMQR and virulence-encoding genes were detected more frequently in cases than in controls. In diarrhoea endemic areas, asymptomatic carriers may play a crucial role in the transmission of multidrug-resistant Shigella spp. with all the putative virulence genes. © 2014 The Authors.

Antibiotics for the treatment of Cholera, Shigella and Cryptosporidium in children.

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Das, Jai K; Ali, Anum; Salam, Rehana A; Bhutta, Zulfiqar A

2013-01-01

Diarrhea is a major contributor to the burden of morbidity and mortality in children; it accounts for a median of 11% of all deaths among children aged less than 5 years, amounting to approximately 0.8 million deaths per year. Currently there is a dearth of literature exploring the effectiveness of antibiotics for diarrhea due to Cholera, Shigella and cryptosporidiosis in children. We reviewed the literature reporting the effect of antibiotics for the treatment of diarrhea due to Cholera, Shigella and Cryptosporidium in children under five years. We used a standardized abstraction and grading format and performed meta-analyses to determine the effect of the treatment with various antibiotics on mortality and rates of clinical and bacteriological/parasitological failure. The CHERG Standard Rules were applied to determine the final effect of treatment with antibiotics on diarrhea morbidity and mortality. For Cholera; the evidence was weak to recommend any effect on mortality. For Shigella; there was no data on mortality; either all-cause or cause specific, hence we used clinical failure rates as a proxy for Shigella deaths and propose that treatment of Shigella dysentery with antibiotics can result in a 82% reduction in diarrhea mortality due to Shigella. For cryptosporidiosis; there was data on all-cause mortality but the evidence was weak hence we used clinical failure rates as a proxy for mortality to estimate that antimicrobial treatment of diarrhea due to cryptosporidiosis can result in a 54% reduction in mortality. There is evidence to recommend antibiotic use for reduction of morbidity and mortality due to Cholera, Shigella and Cryptosporidium. We recommend that more clinical trials should be conducted to evaluate the efficacy and safety of first- and second- line drugs currently in use for treatment for diarrhea and dysentery in both developing and developed countries.

Molecular characterization of Shigella spp. from patients in Gabon 2011-2013.

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Schaumburg, Frieder; Alabi, Abraham S; Kaba, Harry; Lell, Bertrand; Becker, Karsten; Grobusch, Martin P; Kremsner, Peter G; Mellmann, Alexander

2015-04-01

Shigella spp. dysentery is widespread in developing countries; the incidence is particularly high in children between 1-2 years of age. In sub-Saharan Africa, there is a paucity of epidemiological data on Shigella spp., with possible negative consequences for recognition and correct treatment choice for this life-threatening bacterial infection. We therefore characterized Shigella spp. isolates from Gabon. The antimicrobial resistance, virulence factors, genotypes and mobile genetic elements of Shigella isolates (29 S. flexneri; 5 S. boydii; 3 S. sonnei) from a retrospective strain collection were analyzed. High resistance rates were found for gentamicin and tetracycline (100%, 37/37), cotrimoxazole (92%, 34/37) and ampicillin (84%, 31/37). All isolate harbored ial and ipaH; no isolate produced Shiga toxins (stx1/2); enterotoxins (set1A/B) were only found in S. flexneri (n=19). Multilocus sequence types (MLST) clustered with global clones. A high prevalence of atypical class 1 integrons harboring blaOXA30 and aadA1 were detected in S. flexneri, while all S. sonnei carried class 2 integrons. There is a strong link of Gabonese Shigella spp. isolates with pandemic lineages as they cluster with major global clones and frequently carry atypical class 1 integrons which are frequently reported in Shigella spp. from Asia. © The Author 2014. Published by Oxford University Press on behalf of Royal Society of Tropical Medicine and Hygiene. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.

Plants used in Guatemala for the treatment of gastrointestinal disorders. 3. Confirmation of activity against enterobacteria of 16 plants.

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Cáceres, A; Fletes, L; Aguilar, L; Ramirez, O; Figueroa, L; Taracena, A M; Samayoa, B

1993-01-01

Ethnobotanical surveys and literature review identified 408 plants used in Guatemala for the treatment of gastrointestinal disorders. The screening of 84 showed that 34 inhibit one or more enterobacteria; 16 of these were selected for further investigation. Extracts were obtained with three solvents of different polarity (n-hexane, acetone and alcohol) and the in vitro activity was demonstrated against enteropathogenic Escherichia coli, Salmonella enteritidis and Shigella flexneri. The activity of nine ethanolic extracts against enterobacteria, particularly Acalypha guatemalensis, Diphysa robinioides, Lippia dulcis, Psidium guajava and Spondias purpurea was confirmed. The plants with antibacterial activity are discussed.

BACTÉRIAS ENTÉRICAS NAS MÃOS DE MANIPULADORES DE ALIMENTOS DA CIDADE DE ARARAQUARA – SP

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Clarice Queico Fugimura Leite

2007-02-01

Full Text Available Pesquisou-se a presença de bactérias entéricas nas mãos de 48 manipuladores de alimentos de estabelecimentos comerciais da cidade de Araraquara – SP. Verificou-se ausência de Salmonella, Shigella, Yersinia enterocolitica e Escherichia coli enteropatogênica utilizando-se as técnicas propostas. Dos indivíduos analisados 97,9% (47 carreavam outras enterobactérias e 95,8% (46 Streptococcus do grupo D (enterococo. A diversidade da flora bacteriana encontrada evidencia o risco potencial representado pelas mãos na transmissão das infecções alimentares.

Frequency of serovars and antimicrobial resistance in Shigella spp. from Brazil

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Gisele Peirano

2006-05-01

Full Text Available A total of 296 Shigella spp. were received from State Public Health Laboratories, during the period from 1999 to 2004, by National Reference Laboratory for Cholera and Enteric Diseases (NRLCED - IOC/Fiocruz, Rio de Janeiro, Brazil. The frequency of Shigella spp. was: S. flexneri (52.7%, S. sonnei (44.2%, S. boydii (2.3%, and S. dysenteriae (0.6%. The most frequent S. flexneri serovars were 2a and 1b. The highest incidence rates of Shigella isolation were observed in the Southeast (39% and Northeast (34% regions and the lowest rate in the South (3% of Brazil. Strains were further analyzed for antimicrobial susceptibility by disk diffusion method as part of a surveillance program on antimicrobial resistance. The highest rates of antimicrobial resistance were to trimethoprim-sulfamethozaxole (90%, tetracycline (88%, ampicillin (56%, and chloramphenicol (35%. The patterns of antimicrobial resistance among Shigella isolates pose a major difficulty in the determination of an appropriate drug for shigellosis treatment. Continuous monitoring of antimicrobial susceptibilities of Shigella spp. through a surveillance system is thus essential for effective therapy and control measures against shigellosis.

Shigella vaccine development: prospective animal models and current status.

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Kim, Yeon-Jeong; Yeo, Sang-Gu; Park, Jae-Hak; Ko, Hyun-Jeong

2013-01-01

Shigella was first discovered in 1897 and is a major causative agent of dysenteric diarrhea. The number of affected patients has decreased globally because of improved sanitary conditions; however, Shigella still causes serious problems in many subjects, including young children and the elderly, especially in developing countries. Although antibiotics may be effective, a vaccine would be the most powerful solution to combat shigellosis because of the emergence of drug-resistant strains. However, the development of a vaccine is hampered by several problems. First, there is no suitable animal model that can replace human-based studies for the investigation of the in vivo mechanisms of Shigella vaccines. Mouse, guinea pig, rat, rabbit, and nonhuman primates could be used as models for shigellosis, but they do not represent human shigellosis and each has its own weaknesses. However, a recent murine model based on peritoneal infection with virulent S. flexneri 2a is promising. Moreover, although the inflammatory responses and mechanisms such as pathogenassociated molecular patterns and danger-associated molecular patterns have been studied, the pathology and immunology of Shigella are still not clearly defined. Despite these obstacles, many vaccine candidates have been developed, including live attenuated, killed whole cells, conjugated, and subunit vaccines. The development of Shigella vaccines also demands considerations of the cost, routes of administration, ease of storage (stability), cross-reactivity, safety, and immunogenicity. The main aim of this review is to provide a detailed introduction to the many promising vaccine candidates and animal models currently available, including the newly developed mouse model.

Inactivation of Escherichia Coli O157:H7 and Salmonella Enterica on Blueberries in Water Using Ultraviolet Light.

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Liu, Chuhan; Huang, Yaoxin; Chen, Haiqiang

2015-07-01

Ultraviolet light (UV) has antimicrobial effects, but the shadowing effect has limited its application. In this study, a novel setup using UV processing in agitated water was developed to inactivate Escherichia coli O157:H7 and Salmonella on blueberries. Blueberries were dip- or spot-inoculated with E. coli or Salmonella. Blueberries inoculated with E. coli were treated for 2 to 10 min with UV directly (dry UV) or immersed in agitated water during UV treatment (wet UV). E. coli was most easily killed on spot-inoculated blueberries with a 5.2-log reduction after 10-min wet UV treatment. Dip-inoculated blueberries were the most difficult to be decontaminated with only 1.6-log reduction after 10-min wet UV treatment. Wet UV treatment generally showed higher efficacies than dry UV treatment, achieving an average of 1.4 log more reduction for spot-inoculated blueberries. For dip-inoculated blueberries, chlorine washing and UV treatments were less effective, achieving blueberries were UV-treated while being immersed in agitated water containing 100 ppm SDS, 0.5% levulinic acid or 10 ppm chlorine. The 3 chemicals did not significantly enhance the wet UV treatment. Findings of this study suggest that UV treatment could be used as an alternative to chlorine washing for blueberries and potentially for other fresh produce. A novel UV light system for decontamination of blueberries in water was developed and evaluated. Results demonstrated that the decontamination efficacy of this system was generally as effective as chlorine washing, indicating that it could potentially be used as an alternative to chlorine washing for blueberries and other fresh produce. © 2015 Institute of Food Technologists®

Membrane damage and viability loss of thermally treated and high hydrostatic pressurized E. coli 0157:H7 and Salmonella spp. in apple juice

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Differences in membrane damage including leakage of intracellular UV-materials and loss of viability of Salmonella spp. and Escherichia coli O157:H7 bacteria in apple juice following thermal death time disk (TDT) and high hydrostatic pressure treatments were investigated. Salmonella and E. coli O157...

UV-C Adaptation of Shigella: Morphological, Outer Membrane Proteins, Secreted Proteins, and Lipopolysaccharides Effects.

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Chourabi, Kalthoum; Campoy, Susana; Rodriguez, Jesus A; Kloula, Salma; Landoulsi, Ahmed; Chatti, Abdelwaheb

2017-11-01

Water UV disinfection remains extremely important, particularly in developing countries where drinking and reclaimed crop irrigation water may spread devastating infectious diseases. Enteric bacterial pathogens, among which Shigella, are possible contaminants of drinking and bathing water and foods. To study the effect of UV light on Shigella, four strains were exposed to different doses in a laboratory-made irradiation device, given that the ultraviolet radiation degree of inactivation is directly related to the UV dose applied to water. Our results showed that the UV-C rays are effective against all the tested Shigella strains. However, UV-C doses appeared as determinant factors for Shigella eradication. On the other hand, Shigella-survived strains changed their outer membrane protein profiles, secreted proteins, and lipopolysaccharides. Also, as shown by electron microscopy transmission, morphological alterations were manifested by an internal cytoplasm disorganized and membrane envelope breaks. Taken together, the focus of interest of our study is to know the adaptive mechanism of UV-C resistance of Shigella strains.

Prevalence of Salmonella and E. coli in neonatal diarrheic calves

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F.R. El-Seedy

2016-03-01

Full Text Available Neonatal calf diarrhea remains one of the most important problems faced by livestock, causing great economic losses. This study investigated the prevalence of Salmonella and Escherichia coli, especially enterotoxigenic E. coli (ETEC, in diarrheic calves. Fecal samples were collected from 127 diarrheic calves up to 3 months of age at 12 farms from different governorates in Egypt. 119 bacterial isolates (93.7% were recovered and the prevalences of Salmonella and E. coli in diarrheic calves were 18.1% and 75.6%, respectively. Serotyping of Salmonella isolates revealed that S. Enteritidis and S. Typhimurium were the most prevalent serotypes, representing 60.9% and 30.4%, respectively, while S. Dublin was 8.7%. Serogrouping of E. coli isolates showed that 10 O-serogroups were obtained where O26 and O103 were the most prevalent (17.7% of each. Salmonella serotypes showed positive results with PCR test using oligonucleotide primer amplifying 521 bp fragment of invA gene of Salmonella while 70% of E. coli serogroups possessed ETEC virulent gene (K99. The in-vitro antibiotic sensitivity test indicated that Salmonella serotypes showed high sensitivity against enrofloxacin, spectinomycin and neomycin while E. coli isolates showed high sensitivities against marbofloxacin, spectinomycin and neomycin only.

Shigella from Humans in Thailand During 1993 to 2006

DEFF Research Database (Denmark)

Bangtrakulnonth, A.; Wong, Danilo Lo Fo; Vieira, Antonio

2008-01-01

In Thailand during 1993-2006, a total of 9063 Shigella isolates from different medical centers were serotyped and trends over time and. spatial clustering analyzed. Of 3583 cases with age information, 1315 (37%) cases were from children between 0 and 4 years and 684 (1.9%) from children between 5...... and 8 years. Most infections were recorded during 1993-1994 (> 1500 per year), decreasing to Shigella flexneri accounted for 2241 (65%) of 3474 isolations. This proportion decreased to 64 (36%) of 176 infections in 2006....... Most infections occurred during July and August, and fewest in December. S. flexneri clustered around Bangkok, and Shigella sonnei in southern Thailand. Most S. flexneri infections were caused by serotype 2a (1590 of 4035) followed by serotype var X (1249). For both serotypes, a pronounced decrease...

New antimicrobial nitrofuran, trans-5-amino-3-[2-(5-nitro-2-furyl)vinyl]-delta2-1,2,4-oxadiazole: antimicrobial efficacy in mice, rats, and guinea pigs.

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McRipley, R J; Gadebusch, H H; Pansy, F; Semar, R

1974-09-01

A new antimicrobial nitrofuran designated SQ 18,506 showed some therapeutic activity when administered orally to mice infected with Escherichia coli, Salmonella schottmuelleri, Shigella flexneri, or Klebsiella pneumoniae. Animals infected parenterally with Streptococcus pyogenes, Proteus mirabilis, Mycobacterium tuberculosis, and Candida albicans, or topically with Trichophyton mentagrophytes, did not respond to therapy with the drug at the dosage levels used. The compound was as effective as metronidazole in the topical treatment of experimental trichomonal infections in mice and in guinea pigs and as effective as nystatin, candicidin, or a sulfanilamide-aminacrine hydrochloride cream in the treatment of a candidal vaginal infection in rats. The chemotherapeutic efficacy of SQ 18,506 in experimental vaginitis caused by Escherichia coli in the rat surpassed that shown by four commercial products available for the treatment of bacterial vaginitis.

New Antimicrobial Nitrofuran, trans-5-Amino-3-[2-(5-Nitro-2-Furyl)Vinyl]-Δ2 -1,2,4-Oxadiazole: Antimicrobial Efficacy in Mice, Rats, and Guinea pigs

Science.gov (United States)

McRipley, R. J.; Gadebusch, H. H.; Pansy, F.; Semar, R.

1974-01-01

A new antimicrobial nitrofuran designated SQ 18,506 showed some therapeutic activity when administered orally to mice infected with Escherichia coli, Salmonella schottmuelleri, Shigella flexneri, or Klebsiella pneumoniae. Animals infected parenterally with Streptococcus pyogenes, Proteus mirabilis, Mycobacterium tuberculosis, and Candida albicans, or topically with Trichophyton mentagrophytes, did not respond to therapy with the drug at the dosage levels used. The compound was as effective as metronidazole in the topical treatment of experimental trichomonal infections in mice and in guinea pigs and as effective as nystatin, candicidin, or a sulfanilamide-aminacrine hydrochloride cream in the treatment of a candidal vaginal infection in rats. The chemotherapeutic efficacy of SQ 18,506 in experimental vaginitis caused by Escherichia coli in the rat surpassed that shown by four commercial products available for the treatment of bacterial vaginitis. PMID:15830472

Sensitive seminested PCR method for the detection of Shigella in spiked environmental water samples

CSIR Research Space (South Africa)

Theron, J

2001-03-01

Full Text Available to shigellae and EIEC and implicated in virulent functions, is present in multiple copies on the invasion plasmid and the chromosome (Venkatesan et al., 1989, 1991; Hartman et al., 1990; Hale, 1991). The standard procedure for Shigella spp. detection is based... on isolation of Shigella by selective culture media followed by identi?cation by biochemical tests and agglutination assays (Frankel et al., 1989; June et al., 1993). This process may take 48?72 h or even longer to obtain results. Since shigellae are very...

Symptomatic Shigella sonnei urinary tract infection in pregnancy.

Science.gov (United States)

Baka, S; Spathi, A; Tsouma, I; Kouskouni, E

2013-01-01

This report describes a case of urinary tract infection (UTI) due to Shigella sonnei during pregnancy. A 31-year-old pregnant woman was admitted complaining of left-flank tenderness, dysuria, and fever. Following examination, significant laboratory data were collected including increased leukocyte count (10,800/ul with 86% neutrophils) and C-reactive protein (9.6 mg/dl). Urinalysis revealed 30 to 50 leukocytes per high power field while from the quantitative urine culture Shigella sonnei was recovered after 24 h incubation at 37 degrees C. After a two-week course with 750 mg cefuroxime every 8 h, the patient experienced gradual resolution of all symptoms and urinary cultures were negative two weeks and one month, respectively, after completing the therapy. The gestational course was uneventful and the patient delivered a healthy baby girl at term. Shigella sonnei can be responsible for UTI during pregnancy even when no predisposing factors or an apparent source of infection can be identified.

Antibiotic susceptibility profile of bacteria isolated from natural sources of water from rural areas of East Sikkim

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Shubra Poonia

2014-01-01

Full Text Available Background: Contamination of water, food, and environment with antibiotic-resistant bacteria poses a serious public health issue. Objective: The objective was to study the bacterial pollution of the natural sources of water in east Sikkim and to determine the antimicrobial profile of the bacterial isolates. Materials and Methods: A total of 225 samples, 75 each during winter, summer, and monsoon season were collected from the same source in every season for bacteriological analysis by membrane filtration method. Antibiotic susceptibility test was performed using standard disc diffusion method. Results: A total of 19 bacterial species of the genera Escherichia, Klebsiella, Proteus, Salmonella, Shigella, Enterobacter, Citrobacter, Morganella, Pseudomonas, Acinetobacter, Flavobacterium, and Serratia were isolated and their antimicrobial sensitivity tested. Generally, most bacterial isolates except Salmonella and Shigella species were found resistant to commonly used antibiotics such as ampicillin (57.5%, trimethoprim/sulfamethoxaole (39.1%, amoxicillin/clavulanic acid (37.4%, cefixime (34.5%, tetracycline (29.1%, ceftazidime (26.3%, ofloxacin (25.9%, amikacin (8.7%, and gentamicin (2.7% but sensitive to imipenem and piperacillin/tazobactam. Conclusion: Natural sources of water in east Sikkim are grossly contaminated with bacteria including enteropathogens. The consumption of untreated water from these sources might pose health risk to consumers.

Structural Insight inot the low Affinity Between Thermotoga maritima CheA and CheB Compared to their Escherichia coli/Salmonella typhimurium Counterparts

Energy Technology Data Exchange (ETDEWEB)

S Park; B Crane

2011-12-31

CheA-mediated CheB phosphorylation and the subsequent CheB-mediated demethylation of the chemoreceptors are important steps required for the bacterial chemotactic adaptation response. Although Escherichia coli CheB has been reported to interact with CheA competitively against CheY, we have observed that Thermotoga maritima CheB has no detectable CheA-binding. By determining the CheY-like domain crystal structure of T. maritima CheB, and comparing against the T. maritima CheY and Salmonella typhimurium CheB structures, we propose that the two consecutive glutamates in the {beta}4/{alpha}4 loop of T. maritima CheB that is absent in T. maritima CheY and in E. coli/S. typhimurium CheB may be one factor contributing to the low CheA affinity.

Dielectric barrier discharge atmospheric cold plasma inhibits Escherichia coli O157:H7, Salmonella, Listeria monocytogenes, and Tulane virus in Romaine lettuce.

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Min, Sea C; Roh, Si Hyeon; Niemira, Brendan A; Sites, Joseph E; Boyd, Glenn; Lacombe, Alison

2016-11-21

The present study investigated the effects of dielectric barrier discharge atmospheric cold plasma (DACP) treatment on the inactivation of Escherichia coli O157:H7, Salmonella, Listeria monocytogenes, and Tulane virus (TV) on Romaine lettuce, assessing the influences of moisture vaporization, modified atmospheric packaging (MAP), and post-treatment storage on the inactivation of these pathogens. Romaine lettuce was inoculated with E. coli O157:H7, Salmonella, L. monocytogenes (~6logCFU/g lettuce), or TV (~2logPFU/g lettuce) and packaged in either a Petri dish (diameter: 150mm, height: 15mm) or a Nylon/polyethylene pouch (152×254mm) with and without moisture vaporization. Additionally, a subset of pouch-packaged leaves was flushed with O 2 at 5% or 10% (balance N 2 ). All of the packaged lettuce samples were treated with DACP at 34.8kV for 5min and then analyzed either immediately or following post-treatment storage for 24h at 4°C to assess the inhibition of microorganisms. DACP treatment inhibited E. coli O157:H7, Salmonella, L. monocytogenes, and TV by 1.1±0.4, 0.4±0.3, 1.0±0.5logCFU/g, and 1.3±0.1logPFU/g, respectively, without environmental modifications of moisture or gas in the packages. The inhibition of the bacteria was not significantly affected by packaging type or moisture vaporization (p>0.05) but a reduced-oxygen MAP gas composition attenuated the inhibition rates of E. coli O157:H7 and TV. L. monocytogenes continued to decline by an additional 0.6logCFU/g in post-treatment cold storage for 24h. Additionally, both rigid and flexible conventional plastic packages appear to be suitable for the in-package decontamination of lettuce with DACP. Published by Elsevier B.V.

Shigella Sonnei and Shiga Toxin

Centers for Disease Control (CDC) Podcasts

2016-07-28

Katherine Lamba, an infectious disease epidemiologist with the California Department of Public Health, discusses Shiga Toxin producing Shigella sonnei.  Created: 7/28/2016 by National Center for Emerging and Zoonotic Infectious Diseases (NCEZID).   Date Released: 7/28/2016.

Report: Studies on antibacterial activity of some traditional medicinal plants used in folk medicine.

Science.gov (United States)

Israr, Fozia; Hassan, Fouzia; Naqvi, Baqir Shyum; Azhar, Iqbal; Jabeen, Sabahat; Hasan, S M Farid

2012-07-01

Ethanolic extracts of eight medicinal plants commonly used in folk medicine were tested for their antibacterial activity against four Gram positive strains (Bacillus subtilis, Staphylococcus aureus, Staphylococcus epidermidis and, Streptococcus pneumoniae) and six Gram negative strains (Escherichia coli, Proteus vulgaris, Proteus mirabilis. Salmonella typhi para A, Salmonella typhi para B and Shigella dysenteriae) that were obtained from different pathological laboratories located in Karachi, Pakistan. Disc diffusion method was used to analyze antibacterial activity. Out of eight, five medicinal plants showed antibacterial activity against two or more than two microbial species. The most effective antimicrobial plant found to be Punica granatum followed by Curcuma zedoaria Rosc, Grewia asiatica L and Carissa carandas L, Curcuma caesia Roxb respectively. From these results, it is evident that medicinal plants could be used as a potential source of new antibacterial agents.

Molecular characterization of Shigella spp. from patients in Gabon 2011-2013

NARCIS (Netherlands)

Schaumburg, Frieder; Alabi, Abraham S.; Kaba, Harry; Lell, Bertrand; Becker, Karsten; Grobusch, Martin P.; Kremsner, Peter G.; Mellmann, Alexander

2015-01-01

Shigella spp. dysentery is widespread in developing countries; the incidence is particularly high in children between 1-2 years of age. In sub-Saharan Africa, there is a paucity of epidemiological data on Shigella spp., with possible negative consequences for recognition and correct treatment choice

Predicting Salmonella populations from biological, chemical, and physical indicators in Florida surface waters.

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McEgan, Rachel; Mootian, Gabriel; Goodridge, Lawrence D; Schaffner, Donald W; Danyluk, Michelle D

2013-07-01

Coliforms, Escherichia coli, and various physicochemical water characteristics have been suggested as indicators of microbial water quality or index organisms for pathogen populations. The relationship between the presence and/or concentration of Salmonella and biological, physical, or chemical indicators in Central Florida surface water samples over 12 consecutive months was explored. Samples were taken monthly for 12 months from 18 locations throughout Central Florida (n = 202). Air and water temperature, pH, oxidation-reduction potential (ORP), turbidity, and conductivity were measured. Weather data were obtained from nearby weather stations. Aerobic plate counts and most probable numbers (MPN) for Salmonella, E. coli, and coliforms were performed. Weak linear relationships existed between biological indicators (E. coli/coliforms) and Salmonella levels (R(2) Florida surface water through logistic regression.

Imported edible leaves collected at retail sale in England during 2017 with an emphasis on betel and curry leaves: Microbiological quality with respect to Salmonella, Shiga-toxin-producing E. coli (STEC) and levels of Escherichia coli.

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McLauchlin, J; Aird, H; Charlett, A; Chattaway, M; Elviss, N; Hartman, H; Jenkins, C; Jørgensen, F; Larkin, L; Sadler-Reeves, L; Willis, C

2018-05-26

to investigate the microbiological quality of imported fresh leaves on retail-sale during 2017 with respect to Salmonella, Shiga-toxin-producing Escherichia coli (STEC) and levels of E.coli. 279 samples of imported edible leaves (69 banana, 77 betel, 118 curry and 15 other types) were tested. Salmonella spp. which were confirmed by whole genome sequencing and isolated from 44 samples, 26% from curry leaves, 14% from betel and 2.4% from all other leaf types: 80% of all samples contained ≥10 2 , 44% ≥10 3 and 22% ≥10 4 cfu of E.coli cfu/g. All samples where Salmonella were detected also yielded ≥20 cfu of E.coli/g. 54 samples were tested for STEC which was detected in 6 samples and isolated from three: one was identified as STEC O157:H7. this report further highlights an ongoing problem of Salmonella contamination of imported fresh edible leaves. amongst all food tested by Public Health England (approximately 11,000 per annum), curry leaves were the herb most commonly contaminated with Salmonella, and betel leaves were the most commonly contaminated ready-to-eat food. The high proportion with unsatisfactory E. coli levels and the detection of STEC suggests risks of contamination by multiple enteric pathogens. This article is protected by copyright. All rights reserved. This article is protected by copyright. All rights reserved.

Characterization of a novel fusion protein from IpaB and IpaD of Shigella spp. and its potential as a pan-Shigella vaccine.

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Martinez-Becerra, Francisco J; Chen, Xiaotong; Dickenson, Nicholas E; Choudhari, Shyamal P; Harrison, Kelly; Clements, John D; Picking, William D; Van De Verg, Lillian L; Walker, Richard I; Picking, Wendy L

2013-12-01

Shigellosis is an important disease in the developing world, where about 90 million people become infected with Shigella spp. each year. We previously demonstrated that the type three secretion apparatus (T3SA) proteins IpaB and IpaD are protective antigens in the mouse lethal pulmonary model. In order to simplify vaccine formulation and process development, we have evaluated a vaccine design that incorporates both of these previously tested Shigella antigens into a single polypeptide chain. To determine if this fusion protein (DB fusion) retains the antigenic and protective capacities of IpaB and IpaD, we immunized mice with the DB fusion and compared the immune response to that elicited by the IpaB/IpaD combination vaccine. Purification of the DB fusion required coexpression with IpgC, the IpaB chaperone, and after purification it maintained the highly α-helical characteristics of IpaB and IpaD. The DB fusion also induced comparable immune responses and retained the ability to protect mice against Shigella flexneri and S. sonnei in the lethal pulmonary challenge. It also offered limited protection against S. dysenteriae challenge. Our results show the feasibility of generating a protective Shigella vaccine comprised of the DB fusion.

Characterization of a Novel Fusion Protein from IpaB and IpaD of Shigella spp. and Its Potential as a Pan-Shigella Vaccine

Science.gov (United States)

Martinez-Becerra, Francisco J.; Chen, Xiaotong; Dickenson, Nicholas E.; Choudhari, Shyamal P.; Harrison, Kelly; Clements, John D.; Picking, William D.; Van De Verg, Lillian L.; Walker, Richard I.

2013-01-01

Shigellosis is an important disease in the developing world, where about 90 million people become infected with Shigella spp. each year. We previously demonstrated that the type three secretion apparatus (T3SA) proteins IpaB and IpaD are protective antigens in the mouse lethal pulmonary model. In order to simplify vaccine formulation and process development, we have evaluated a vaccine design that incorporates both of these previously tested Shigella antigens into a single polypeptide chain. To determine if this fusion protein (DB fusion) retains the antigenic and protective capacities of IpaB and IpaD, we immunized mice with the DB fusion and compared the immune response to that elicited by the IpaB/IpaD combination vaccine. Purification of the DB fusion required coexpression with IpgC, the IpaB chaperone, and after purification it maintained the highly α-helical characteristics of IpaB and IpaD. The DB fusion also induced comparable immune responses and retained the ability to protect mice against Shigella flexneri and S. sonnei in the lethal pulmonary challenge. It also offered limited protection against S. dysenteriae challenge. Our results show the feasibility of generating a protective Shigella vaccine comprised of the DB fusion. PMID:24060976

The Inside Story of Shigella Invasion of Intestinal Epithelial Cells

Science.gov (United States)

Carayol, Nathalie; Tran Van Nhieu, Guy

2013-01-01

As opposed to other invasive pathogens that reside into host cells in a parasitic mode, Shigella, the causative agent of bacillary dysentery, invades the colonic mucosa but does not penetrate further to survive into deeper tissues. Instead, Shigella invades, replicates, and disseminates within the colonic mucosa. Bacterial invasion and spreading in intestinal epithelium lead to the elicitation of inflammatory responses responsible for the tissue destruction and shedding in the environment for further infection of other hosts. In this article, we highlight specific features of the Shigella arsenal of virulence determinants injected by a type III secretion apparatus (T3SA) that point to the targeting of intestinal epithelial cells as a discrete route of invasion during the initial event of the infectious process. PMID:24086068

Prevalence and counts of Salmonella spp. in minimally processed vegetables in São Paulo, Brazil.

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Sant'Ana, Anderson S; Landgraf, Mariza; Destro, Maria Teresa; Franco, Bernadette D G M

2011-09-01

Minimally processed vegetables (MPV) may be important vehicles of Salmonella spp. and cause disease. This study aimed at detecting and enumerating Salmonella spp. in MPV marketed in the city of São Paulo, Brazil. A total of 512 samples of MPV packages collected in retail stores were tested for Salmonella spp. and total coliforms and Escherichia coli as indication of the hygienic status. Salmonella spp. was detected in four samples, two using the detection method and two using the counting method, where the results were 8.8 × 10(2) CFU/g and 2.4 × 10(2) CFU/g. The serovars were Salmonella Typhimurium (three samples) and Salmonella enterica subsp. enterica O:47:z4,z23:- (one sample). Fourteen samples (2.7%) presented counts of E. coli above the maximum limit established by the Brazilian regulation for MPV (10(2) CFU/g). Therefore, tightened surveillance and effective intervention strategies are necessary in order to address consumers and governments concerns on safety of MPV. Copyright © 2011 Elsevier Ltd. All rights reserved.

A Review of Current Research and Knowledge Gaps in the Epidemiology of Shiga Toxin-Producing Escherichia coli and Salmonella spp. in Trinidad and Tobago

Directory of Open Access Journals (Sweden)

Anil K. Persad

2018-04-01

Full Text Available Salmonella and Shiga toxin-producing Escherichia coli are two of the main causes of foodborne disease globally, and while they have been implicated as possible causes of foodborne disease within the Caribbean region, the actual incidence is unknown. Trinidad and Tobago, one of the larger countries in the Caribbean, has an estimated annual foodborne disease burden of over 100,000 cases and, similar to other countries, the etiology of most of these cases is unknown. Both pathogens can reside as part of the normal gastrointestinal microflora of many wild and domestic animals, with animals acting as reservoirs, spillover hosts, or dead-end hosts. Carriage in animal species can be asymptomatic or, in the case of Salmonella in particular, there may be clinical manifestation in animals, which resemble the disease seen in humans. In this review, we will focus on the epidemiology of these two foodborne pathogens in Trinidad and Tobago and identify any knowledge gaps in the published literature. The filling of this critical knowledge void is essential for the development and implementation of appropriate mechanisms to reduce the dissemination and transmission of these pathogens, not only in Trinidad and Tobago, but also in the wider Caribbean.

In Vitro Assembly of the Outer Core of the Lipopolysaccharide from Escherichia coli K-12 and Salmonella typhimurium

Science.gov (United States)

2015-01-01

There are five distinct core structures in the lipopolysaccharides of Escherichia coli and at least two in Salmonella isolates, which vary principally in the outer core oligosaccharide. Six outer core glycosyltransferases, E. coli K-12 WaaG, WaaB, and WaaO and Salmonella typhimurium WaaI, WaaJ, and WaaK, were cloned, overexpressed, and purified. A novel substrate for WaaG was isolated from ΔwaaG E. coli overexpressing the lipid A phosphatase lpxE and the lipid A late acyltransferase lpxM. The action of lpxE and lpxM in the ΔwaaG background yielded heptose2-1-dephospho Kdo2-lipid A, a 1-dephosphorylated hexa-acylated lipid A with the inner core sugars that is easily isolated by organic extraction. Using this structurally defined acceptor and commercially available sugar nucleotides, each outer core glycosyltransferases was assayed in vitro. We show that WaaG and WaaB add a glucose and galactose sequentially to heptose2-1-dephospho Kdo2-lipid A. E. coli K-12 WaaO and S. typhimurium WaaI add a galactose to the WaaG/WaaB product but can also add a galactose to the WaaG product directly without the branched core sugar added by WaaB. Both WaaI and WaaO require divalent metal ions for optimal activity; however, WaaO, unlike WaaI, can add several glucose residues to its lipid acceptor. Using the product of WaaG, WaaB, and WaaI, we show that S. typhimurium WaaJ and WaaK transfer a glucose and N-acetylglucosamine, respectively, to yield the full outer core. This is the first demonstration of the in vitro assembly of the outer core of the lipopolysaccharide using defined lipid A-oligosaccharide acceptors and sugar donors. PMID:24479701

Shigella sonnei and hemolytic uremic syndrome: A case report and literature review

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Casey Adams

2017-01-01

Full Text Available Hemolytic uremic syndrome (HUS is a well-described process that is known to cause severe renal dysfunction, thrombocytopenia, and anemia. HUS is typically associated with toxins (shiga-like and shigella toxin found in strains of E. coli and Shigella spp [1–3]. We present a case of a 27 year-old man with jaundice, thrombocytopenia, and renal dysfunction who was found to have HUS in the setting of Shigella sonnei infection. Outside of developing countries, cases of HUS related to S. sonnei are largely unreported.

A novel enterocin T1 with anti-Pseudomonas activity produced by Enterococcus faecium T1 from Chinese Tibet cheese.

Science.gov (United States)

Liu, Hui; Zhang, Lanwei; Yi, Huaxi; Han, Xue; Gao, Wei; Chi, Chunliang; Song, Wei; Li, Haiying; Liu, Chunguang

2016-02-01

An enterocin-producing Enterococcus faecium T1 was isolated from Chinese Tibet cheese. The enterocin was purified by SP-Sepharose and reversed phase HPLC. It was identified as unique from other reported bacteriocins based on molecular weight (4629 Da) and amino acid compositions; therefore it was subsequently named enterocin T1. Enterocin T1 was stable at 80-100 °C and over a wide pH range, pH 3.0-10.0. Protease sensitivity was observed to trypsin, pepsin, papain, proteinase K, and pronase E. Importantly, enterocin T1 was observed to inhibit the growth of numerous Gram-negative and Gram-positive bacteria including Pseudomonas putida, Pseudomonas aeruginosa, Pseudomonas fluorescens, Escherichia coli, Salmonella typhimurium, Shigella flexneri, Shigella sonnei, Staphylococcus aureus, Listeria monocytogenes. Take together, these results suggest that enterocin T1 is a novel bacteriocin with the potential to be used as a bio-preservative to control Pseudomonas spp. in food.

Antimicrobial resistant Salmonella enterica and Escherichia coli recovered from dairy operations

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Antimicrobial resistance has become a major public health concern and animal agriculture is often implicated as a source of resistant bacteria. The primary objective of this study was to determine prevalence of antimicrobial resistance in Salmonella and E. coli from healthy animals on dairy farms i...

[Detection of CRISPR and its relationship to drug resistance in Shigella].

Science.gov (United States)

Wang, Linlin; Wang, Yingfang; Duan, Guangcai; Xue, Zerun; Guo, Xiangjiao; Wang, Pengfei; Xi, Yuanlin; Yang, Haiyan

2015-04-04

To detect clustered regularly interspaced short palindromic repeats (CRISPR) in Shigella, and to analyze its relationship to drug resistance. Four pairs of primers were used for the detection of convincing CRISPR structures CRISPR-S2 and CRISPR-S4, questionable CRISPR structures CRISPR-S1 and CRISPR-S3 in 60 Shigella strains. All primers were designed using sequences in CRISPR database. CRISPR Finder was used to analyze CRISPR and susceptibilities of Shigella strains were tested by agar diffusion method. Furthermore, we analyzed the relationship between drug resistance and CRISPR-S4. The positive rate of convincing CRISPR structures was 95%. The four CRISPR loci formed 12 spectral patterns (A-L), all of which contained convincing CRISPR structures except type K. We found one new repeat and 12 new spacers. The multi-drug resistance rate was 53. 33% . We found no significant difference between CRISPR-S4 and drug resistant. However, the repeat sequence of CRISPR-S4 in multi- or TE-resistance strains was mainly R4.1 with AC deletions in the 3' end, and the spacer sequences of CRISPR-S4 in multi-drug resistance strains were mainly Sp5.1, Sp6.1 and Sp7. CRISPR was common in Shigella. Variations df repeat sequences and diversities of spacer sequences might be related to drug resistance in Shigella.

Attachment behaviour of Escherichia coli K12 and Salmonella Typhimurium P6on food contact surfaces for food transportation

DEFF Research Database (Denmark)

Abban, Stephen; Jakobsen, Mogens; Jespersen, Lene

2012-01-01

The role of cargo container lining materials aluminium, a fibre reinforced plastic (FRP) and stainless steel in bacterial cross contamination during transport was assessed. For this, attachment and detachment of Escherichia coli K12 and Salmonella Typhimurium P6 on the three surfaces in the absence....... Typhimurium P6 respectively. Correlation with roughness average was poor; r = -0.425 and -0.413 respectively for E. coli K12 and S. Typhimurium P6. Presence of residue caused significant reduction (p ... material sections of the same surfaces. We report these observations for the first time for aluminium and the FRP material and in part for stainless steel. The S. Typhimurium P6 strain also had significantly higher level of attachment than the E. coli K12 strain. Our findings show that food residue...

Occurrence of integrons and resistance genes among sulphonamide-resistant Shigella spp. from Brazil

DEFF Research Database (Denmark)

Peirano, G.; Agersø, Yvonne; Aarestrup, Frank Møller

2005-01-01

Objectives: To determine the occurrence of class 1 and 2 integrons and antimicrobial resistance genes among sulphonamide-resistant Shigella strains isolated in Brazil during 1999-2003. Methods: Sixty-two Shigella (Shigella flexneri, n = 47 and Shigella sonnei, n = 15) were tested against 21...... antimicrobial agents. The presence of integrons classes 1 and 2 and antimicrobial resistance genes was investigated by PCR using specific primers. Results: A total of eight antimicrobial resistance profiles were identified, with the profile of resistance to sulfamethoxazole, trimethoprim, spectinomycin...... of 2214 bp harbouring a gene cassette array conferring resistance to trimethoprim, streptothricin and spectinomycin/streptomycin. The genes coding for resistance to chloramphenicol (catA1), tetracycline [tet(A) and tet(B)] and ampicillin (bla(OXA) and bla(TEM)), were detected in resistant strains...

SOS gene induction and possible mutagenic effects of freeze-drying in Escherichia coli and Salmonella typhimurium.

Science.gov (United States)

Rosen, Rachel; Buchinger, Sebastian; Pfänder, Ramona; Pedhazur, Rami; Reifferscheid, Georg; Belkin, Shimshon

2016-11-01

We report the results of a study of the potential negative effects of the freeze-drying process, normally considered a benign means for long-term conservation of living cells and the golden standard in bacterial preservation. By monitoring gene induction using a whole-cell Escherichia coli bioreporter panel, in which diverse stress-responsive gene promoters are fused to luminescent or fluorescent reporting systems, we have demonstrated that DNA repair genes belonging to the SOS operon (recA, sulA, uvrA, umuD, and lexA) were induced upon resuscitation from the freeze-dried state, whereas other stress-responsive promoters such as grpE, katG, phoA, soxS, and sodA were not affected. This observation was confirmed by the UMU-chromotest (activation of the umuD gene promoter) in Salmonella typhimurium, as well as by real-time PCR analyses of selected E. coli SOS genes. We further show that a functional SOS operon is important in viability maintenance following resuscitation, but that at the same time, this repair system may introduce significantly higher mutation rates, comparable to those induced by high concentrations of a known mutagen. Our results also indicate that the entire freeze-drying process, rather than either freezing or drying separately, is instrumental in the induction of DNA damage.

Escherichia coli O157:H7 induces stronger plant immunity than Salmonella enterica Typhimurium SL1344.

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Roy, Debanjana; Panchal, Shweta; Rosa, Bruce A; Melotto, Maeli

2013-04-01

Consumption of fresh produce contaminated with bacterial human pathogens has resulted in various, sometimes deadly, disease outbreaks. In this study, we assessed plant defense responses induced by the fully pathogenic bacteria Escherichia coli O157:H7 and Salmonella enterica serovar Typhimurium SL1344 in both Arabidopsis thaliana and lettuce (Lactuca sativa). Unlike SL1344, O157:H7 induced strong plant immunity at both pre-invasion and post-invasion steps of infection. For instance, O157:H7 triggered stomatal closure even under high relative humidity, an environmental condition that generally weakens plant defenses against bacteria in the field and laboratory conditions. SL1344 instead induced a transient stomatal immunity. We also observed that PR1 gene expression was significantly higher in Arabidopsis leaves infected with O157:H7 compared with SL1344. These results suggest that plants may recognize and respond to some human pathogens more effectively than others. Furthermore, stomatal immunity can diminish the penetration of human pathogens through the leaf epidermis, resulting in low bacterial titers in the plant apoplast and suggesting that additional control measures can be employed to prevent food contamination. The understanding of how plant responses can diminish bacterial contamination is paramount in preventing outbreaks and improving the safety of food supplies.

Survival or growth of inoculated Escherichia coli O157:H7 and Salmonella on yellow onions (Allium cepa) under conditions simulating food service and consumer handling and storage.

Science.gov (United States)

Lieberman, Vanessa M; Zhao, Irene Y; Schaffner, Donald W; Danyluk, Michelle D; Harris, Linda J

2015-01-01

Whole and diced yellow onions (Allium cepa) were inoculated with five-strain cocktails of rifampin-resistant Escherichia coli O157:H7 or Salmonella and stored under conditions to simulate food service or consumer handling. The inoculum was grown in broth (for both whole and diced onion experiments) or on agar plates (for whole onion experiments). Marked circles (3.3 cm in diameter) on the outer papery skin of whole onions were spot inoculated (10 μl in 10 drops) at 7 log CFU per circle, and onions were stored at 4°C, 30 to 50 % relative humidity, or at ambient conditions (23°C, 30 to 50 % relative humidity). Diced onions were inoculated at 3 log CFU/g and then stored in open or closed containers at 4°C or ambient conditions. Previously inoculated and ambient-stored diced onions were also mixed 1:9 (wt/wt) with refrigerated uninoculated freshly diced onions and stored in closed containers at ambient conditions. Inoculated pathogens were recovered in 0.1 % peptone and plated onto selective and nonselective media supplemented with 50 μg/ml rifampin. Both E. coli O157:H7 and Salmonella populations declined more rapidly on onion skins when the inoculum was prepared in broth rather than on agar. Agar-prepared E. coli O157:H7 and Salmonella declined by 0.4 and 0.3 log CFU per sample per day, respectively, at ambient conditions; at 4°C the rates of reduction were 0.08 and 0.06 log CFU per sample per day for E. coli O157:H7 and Salmonella, respectively. Populations of E. coli O157:H7 and Salmonella did not change over 6 days of storage at 4°C in diced onions. Lag times of 6 to 9 h were observed with freshly inoculated onion at ambient conditions; no lag was observed when previously inoculated and uninoculated onions were mixed. Growth rates at ambient conditions were 0.2 to 0.3 log CFU/g/h for E. coli O157:H7 and Salmonella in freshly inoculated onion and 0.2 log CFU/g/h in mixed product. Diced onions support pathogen growth and should be kept refrigerated.

Multidrug Resistance in Infants and Children

Directory of Open Access Journals (Sweden)

Gian Maria Pacifici

2018-02-01

Full Text Available Bacterial infections may cause disease and death. Infants and children are often subject to bacterial infections. Antimicrobials kill bacteria protecting the infected patients andreducing the risk of morbidity and mortality caused by bacteria. The antibiotics may lose their antibacterial activity when they become resistant to a bacteria. The resistance to different antibiotics in a bacteria is named multidrug-resistance. Gram-negative bacilli, especially Escherichia coli, Klebsiella, Enterobacter, Salmonella, Shigella, Pseudomonas, Streptococcus, and Haemophilus influenzae type b, may become resistant. Amikacin ampicillin, amoxicillin, amoxiclav, cefuroxime, cefotaxime, ceftazidime, cefoperazone tetracycline, chloramphenicol, ciprofloxacin, and gentamicin may cause bacterial-resistance. Resistance to bacteria for several pathogens makes complications in the treatment of infections caused by them. Salmonella strains may become resistant to ampicillin, cephalotin, ceftriaxone, gentamicin, amikacin, trimethoprim-sulfamethoxazole, chloramphenicol, and tetracycline. Shigella strains may become resistant to ampicillin, cotrimoxazole, chloramphenicol, and streptomycin. Multidrug-resistance of Streptococcus pneumoniae may be due to β-lactams, macrolides, tetracycline, chloramphenicol, and trimethoprim-sulfamethoxazole. Multidrug-resistance of Pseudomonas aeruginosa may become resistant to β-lactams, chloramphenicol, trimethoprim-sulfamethoxazole, and tetracycline. The antibacterial activity against Haemophilus strains may occur with ampicillin, sulbactam-ampicillin, trimethoprim-sulfamethoxazole, gentamicin, chloramphenicol, and ciprofloxacin. Multidrug-resistance of the Klebsiella species may be due with ampicillin, cefotaxime, cefuroxime, co-amxilav, mezlocillin, chloramphenicol, gentamicin, and ceftazidime. Multidrug-resistance of Escherichia coli may be caused by ampicillin, cotrimoxazole, chloramphenicol, ceftriaxone, and ceftazidime. Vibrio

Shigella Infections in Household Contacts of Pediatric Shigellosis Patients in Rural Bangladesh.

Science.gov (United States)

George, Christine Marie; Ahmed, Shahnawaz; Talukder, Kaisar A; Azmi, Ishrat J; Perin, Jamie; Sack, R Bradley; Sack, David A; Stine, O Colin; Oldja, Lauren; Shahnaij, Mohammad; Chakraborty, Subhra; Parvin, Tahmina; Bhuyian, Sazzadul Islam; Bouwer, Edward; Zhang, Xiaotong; Hasan, Trisheeta N; Luna, Sharmin J; Akter, Fatema; Faruque, Abu S G

2015-11-01

To examine rates of Shigella infections in household contacts of pediatric shigellosis patients, we followed contacts and controls prospectively for 1 week after the index patient obtained care. Household contacts of patients were 44 times more likely to develop a Shigella infection than were control contacts (odds ratio 44.7, 95% CI 5.5-361.6); 29 (94%) household contacts of shigellosis patients were infected with the same species and serotype as the index patient's. Pulsed-field gel electrophoresis showed that 14 (88%) of 16 with infected contacts had strains that were indistinguishable from or closely related to the index patient's strain. Latrine area fly counts were higher in patient households compared with control households, and 2 patient household water samples were positive for Shigella. We show high susceptibility of household contacts of shigellosis patients to Shigella infections and found environmental risk factors to be targeted in future interventions.

Cellular Aspects of Shigella Pathogenesis: Focus on the Manipulation of Host Cell Processes.

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Killackey, Samuel A; Sorbara, Matthew T; Girardin, Stephen E

2016-01-01

Shigella is a Gram-negative bacterium that is responsible for shigellosis. Over the years, the study of Shigella has provided a greater understanding of how the host responds to bacterial infection, and how bacteria have evolved to effectively counter the host defenses. In this review, we provide an update on some of the most recent advances in our understanding of pivotal processes associated with Shigella infection, including the invasion into host cells, the metabolic changes that occur within the bacterium and the infected cell, cell-to-cell spread mechanisms, autophagy and membrane trafficking, inflammatory signaling and cell death. This recent progress sheds a new light into the mechanisms underlying Shigella pathogenesis, and also more generally provides deeper understanding of the complex interplay between host cells and bacterial pathogens in general.

Cladophora (Chlorophyta) spp. harbor human bacterial pathogens in nearshore water of Lake Michigan

Science.gov (United States)

Ishii, S.; Yan, T.; Shively, D.A.; Byappanahalli, M.N.; Whitman, R.L.; Sadowsky, M.J.

2006-01-01

Cladophora glomerata, a macrophytic green alga, is commonly found in the Great Lakes, and significant accumulations occur along shorelines during the summer months. Recently, Cladophora has been shown to harbor high densities of the fecal indicator bacteria Escherichia coli and enterococci. Cladophora may also harbor human pathogens; however, until now, no studies to address this question have been performed. In the present study, we determined whether attachedCladophora, obtained from the Lake Michigan and Burns Ditch (Little Calumet River, Indiana) sides of a breakwater during the summers of 2004 and 2005, harbored the bacterial pathogens Shiga toxin-producing Escherichia coli (STEC),Salmonella, Shigella, and Campylobacter. The presence of potential pathogens and numbers of organisms were determined by using cultural methods and by using conventional PCR, most-probable-number PCR (MPN-PCR), and quantitative PCR (QPCR) performed with genus- and toxin-specific primers and probes. WhileShigella and STEC were detected in 100% and 25%, respectively, of the algal samples obtained near Burns Ditch in 2004, the same pathogens were not detected in samples collected in 2005. MPN-PCR and QPCR allowed enumeration of Salmonella in 40 to 80% of the ditch- and lakeside samples, respectively, and the densities were up to 1.6 × 103 cells per g Cladophora. Similarly, these PCR methods allowed enumeration of up to 5.4 × 102 Campylobacter cells/gCladophora in 60 to 100% of lake- and ditchside samples. The Campylobacterdensities were significantly higher (P fingerprint analyses indicated that genotypically identical Salmonella isolates were associated with geographically and temporally distinct Cladophora samples. However, Campylobacter isolates were genetically diverse. Since animal hosts are thought to be the primary habitat forCampylobacter and Salmonella species, our results suggest that Cladophora is a likely secondary habitat for pathogenic

Antidiarrhoeal and antimicrobial activity of Calpurnia aurea leaf extract

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Umer Shemsu

2013-01-01

Full Text Available Abstract Background In Ethiopia, Calpurnia aurea is used for the treatment of syphilis, malaria, rabies, diabetes, hypertension, diarrhoea, leishmaniasis, trachoma, elephantiasis, fungal diseases and different swellings. However, despite its traditional usage as an antidiarrhoeal and antimicrobial agent, there is limited or no information regarding its effectiveness and mode of action in diarrhoea which may be caused by Shigella flexneri, Staphylococcus aureus, Escherichia coli and Salmonella typhi. Hence, we evaluated the 80% methanol (MeOH extract of dried and powdered leaves of C. aurea for its antidiarrhoeal and antimicrobial activities. Methods Swiss albino mice of either sex were divided into five groups (five/group: Group I served as control and received vehicle (1% Tween 80 at a dose of 10 ml/kg orally; Group II served as standard and received loperamide at the dose of 3 mg/kg orally; Groups III, IV and V served as test groups and received the 80% MeOH leaf extract of C. aurea at doses of 100, 200 and 400 mg/kg orally, respectively. Diarrhoea was induced by oral administration of 0.5 ml castor oil to each mouse, 1 h after the above treatments. During an observation period of 4 h, time of onset of diarrhea, total number of faecal output (frequency of defecation and weight of faeces excreted by the animals were recorded. Data were analyzed using one way analysis of variance followed by Tukey post test. Antimicrobial activity test was conducted using agar well diffusion assay. Clinical isolates tested were Salmonella typhi, Salmonella paratyphi, Salmonella typhimurium, Shigella species, Pseudomonas aeruginosa, Staphylococcus aureus and Escherichia coli. Results In castor oil induced diarrhea model, the 80% methanol leaf extract of C. aurea at 100, 200 and 400 mg/kg and the standard drug loperamide (3 mg/kg significantly reduced the time of onset of diarrhea, the frequency of defecation (total number of faecal output and weight of faeces. C

Prevalence of Salmonella enterica, Listeria monocytogenes, and pathogenic Escherichia coli in bulk tank milk and milk filters from US dairy operations in the National Animal Health Monitoring System Dairy 2014 study.

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Sonnier, Jakeitha L; Karns, Jeffrey S; Lombard, Jason E; Kopral, Christine A; Haley, Bradd J; Kim, Seon-Woo; Van Kessel, Jo Ann S

2018-03-01

The dairy farm environment is a well-documented reservoir for zoonotic pathogens such as Salmonella enterica, Shiga-toxigenic Escherichia coli, and Listeria monocytogenes, and humans may be exposed to these pathogens via consumption of unpasteurized milk and dairy products. As part of the National Animal Health Monitoring System Dairy 2014 study, bulk tank milk (BTM, n = 234) and milk filters (n = 254) were collected from a total of 234 dairy operations in 17 major dairy states and analyzed for the presence of these pathogens. The invA gene was detected in samples from 18.5% of operations and Salmonella enterica was isolated from 18.0% of operations. Salmonella Dublin was detected in 0.7% of operations. Sixteen Salmonella serotypes were isolated, and the most common serotypes were Cerro, Montevideo, and Newport. Representative Salmonella isolates (n = 137) were tested against a panel of 14 antimicrobials. Most (85%) were pansusceptible; the remaining were resistant to 1 to 9 antimicrobials, and within the resistant strains the most common profile was resistance to ampicillin/clavulanic acid, ampicillin, cefoxitin, ceftiofur, ceftriaxone, chloramphenicol, streptomycin, sulfisoxazole, and tetracycline. Listeria spp. were isolated from 19.9% of operations, and L. monocytogenes was isolated from 3.0% of operations. Serogroups 1/2a and 1/2b were the most common, followed by 4b and 4a. One or more E. coli virulence genes were detected in the BTM from 30.5% of operations and in the filters from 75.3% of operations. A combination of stx 2 , eaeA, and γ-tir genes was detected in the BTM from 0.5% of operations and in the filters from 6.6% of operations. The results of this study indicate an appreciable prevalence of bacterial pathogens in BTM and filters, including serovars known to infect humans. Copyright © 2018 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.

Antimicrobial screening of impatiens bicolor royle

International Nuclear Information System (INIS)

Nisar, M.; Ali, I.; Qayum, M.; Kaleem, W.A.; Shah, R.M.; Zia-ul-Haq, M.

2010-01-01

Extracts of Impatiens bicolor Royle obtained from n-hexane (A); dichloromethane (B), ethyl acetate (C), n-butanol (D), aqueous (E) as well as crude (F) were tested In vitro for their antibacterial and antifungal activities. Antibacterial study performed against 6 bacteria viz., Escherichia coli, Bacillus subtilis, Shigella flexenari, Staphylococcus aureus, Pseudomonas aeruginosa and Salmonella typhi indicated that crude and its fractions had no activity at all against any microorganism. The antifungal activity of these extracts was performed against 6 fungi viz., Trichophyton longifusus, Candida albicans, Aspergilus flavus, Microsporum canis, Fusarium solani and Candida glaberata. The extracts showed moderate activity against different fungal strains. (author)

Salmonella

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... Compartir Find out about Salmonella infections linked to Kellogg’s Honey Smacks Cereal Find out about Salmonella infections ... Outbreaks Multistate Outbreak of Salmonella Infections Linked to Kellogg’s Honey Smacks Cereal Multistate Outbreak of Salmonella Adelaide ...

Shigella reroutes host cell central metabolism to obtain high-flux nutrient supply for vigorous intracellular growth.

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Kentner, David; Martano, Giuseppe; Callon, Morgane; Chiquet, Petra; Brodmann, Maj; Burton, Olga; Wahlander, Asa; Nanni, Paolo; Delmotte, Nathanaël; Grossmann, Jonas; Limenitakis, Julien; Schlapbach, Ralph; Kiefer, Patrick; Vorholt, Julia A; Hiller, Sebastian; Bumann, Dirk

2014-07-08

Shigella flexneri proliferate in infected human epithelial cells at exceptionally high rates. This vigorous growth has important consequences for rapid progression to life-threatening bloody diarrhea, but the underlying metabolic mechanisms remain poorly understood. Here, we used metabolomics, proteomics, and genetic experiments to determine host and Shigella metabolism during infection in a cell culture model. The data suggest that infected host cells maintain largely normal fluxes through glycolytic pathways, but the entire output of these pathways is captured by Shigella, most likely in the form of pyruvate. This striking strategy provides Shigella with an abundant favorable energy source, while preserving host cell ATP generation, energy charge maintenance, and survival, despite ongoing vigorous exploitation. Shigella uses a simple three-step pathway to metabolize pyruvate at high rates with acetate as an excreted waste product. The crucial role of this pathway for Shigella intracellular growth suggests targets for antimicrobial chemotherapy of this devastating disease.

5-Nitroimidazole Derivatives and their Antimicrobial Activity

International Nuclear Information System (INIS)

Khan, K.M.; Salar, U.; Yousuf, S.; Naz, F.

2016-01-01

5-Nitroimidazole derivatives 2-8 were synthesized from secnidazole. The syntheses were accomplished in two steps which start from the oxidation of secnidazole to the secnidazolone 1. Secnidazolone 1 was converted into its hydrazone derivative 2-8 by treating with different substituted acid hydrazide. Compounds 2-8 were evaluated for their antimicrobial activity against Gram-positive and Gram-negative bacteria, compounds 3 and 4 showed the significant activity against Staphylococcus epidermidis, however, compound 2 showed good inhibitions against Corynebacterium diphtheria when compared with the standard. Compound 3 showed good inhibitory potential against tested Gram-negative bacterial strains i.e. Enterobacter aerogene, Escherichia coli, Salmonella typhi, Salmonella paratyphi A, Shigella flexeneri and Vibrio choleriae. All synthetic derivatives were also tested against eight fungal stains, however, they were weekly active against Aspergillus flavus and Candida albican. The synthesized compounds were characterized by different spectroscopy techniques. (author)

An integrated approach for finding overlooked genes in Shigella.

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Junping Peng

Full Text Available BACKGROUND: The completion of numerous genome sequences introduced an era of whole-genome study. However, many genes are missed during genome annotation, including small RNAs (sRNAs and small open reading frames (sORFs. In order to improve genome annotation, we aimed to identify novel sRNAs and sORFs in Shigella, the principal etiologic agents of bacillary dysentery. METHODOLOGY/PRINCIPAL FINDINGS: We identified 64 sRNAs in Shigella, which were experimentally validated in other bacteria based on sequence conservation. We employed computer-based and tiling array-based methods to search for sRNAs, followed by RT-PCR and northern blots, to identify nine sRNAs in Shigella flexneri strain 301 (Sf301 and 256 regions containing possible sRNA genes. We found 29 candidate sORFs using bioinformatic prediction, array hybridization and RT-PCR verification. We experimentally validated 557 (57.9% DOOR operon predictions in the chromosomes of Sf301 and 46 (76.7% in virulence plasmid.We found 40 additional co-expressed gene pairs that were not predicted by DOOR. CONCLUSIONS/SIGNIFICANCE: We provide an updated and comprehensive annotation of the Shigella genome. Our study increased the expected numbers of sORFs and sRNAs, which will impact on future functional genomics and proteomics studies. Our method can be used for large scale reannotation of sRNAs and sORFs in any microbe with a known genome sequence.

Serologic reactions against Salmonella in samples from broiler parent stock with and without preceding colibacillosis: A case-control study

DEFF Research Database (Denmark)

Gradel, K.O.; Feld, Niels Christian; Andersen, J. S.

2001-01-01

In the Danish Salmonella Control Program, eggs from broiler parent flocks are surveyed by serologic analysis every 4 wk for antibodies against Salmonella lipopolysaccharide O-antigens 1, 4, 5, 9, and 12 (Mix-enzyme-linked immunosorbent assay [ELISA]) and 6 and 7 (Infantis-ELISA). The antibody...... response is measured in percentage optical density (OD%) of a strong positive reaction, and the cutoff value has been determined to be 40 OD%. Two or more reactors above 40 OD% will place the parent flock under suspicion. There has been concern about possible cross-reactions between Salmonella spp....... and other Enterobacteriaceae, e.g., Escherichia coli, because a high specificity of a Salmonella antibody test is desirable. Moreover, false-positive Salmonella results have economic consequences and impede planning the production. A case-control study based on cases of clinical E. coli infections...

Inactivation of Escherichia coli O157:H7 and Salmonella deposited on gloves in a liquid state and subjected to drying conditions.

Science.gov (United States)

Erickson, Marilyn C; Liao, Jye-Yin; Webb, Cathy C; Habteselassie, Mussie Y; Cannon, Jennifer L

2018-02-02

Gloves are worn by workers harvesting ready-to-eat produce as a deterrent for contaminating the produce with enteric pathogens that may reside on their hands. As fields are not sterile environments, the probability for gloves to become contaminated still exists and therefore it is critical to understand the conditions that affect the survival of pathogens on gloves. Both Escherichia coli O157:H7 and Salmonella deposited on glove surfaces in a liquid state survived longer when the pathogen had been suspended in lettuce sap than when suspended in water. Despite this protection, pathogens deposited on clean single-use gloves were more likely to survive during drying than pathogens deposited on dirty gloves (a film of lettuce sap had been applied to the surface prior to pathogen application and soil had been ground into the gloves). Survival of both E. coli O157:H7 and Salmonella was biphasic with the greatest losses occurring during the first hour of drying followed by much slower losses in the ensuing hours. Pathogens grown in rich media (tryptic soy broth) versus minimal media (M9) as well as those cultured on solid agar versus liquid broth were also more likely to be resistant to desiccation when deposited onto gloves. Although survival of E. coli O157:H7 on nitrile gloves was in general greater than it was on latex gloves, the relative survival of Salmonella on the two glove types was inconsistent. Due to these inconsistencies, no one glove type is considered better than another in reducing the risk for contamination with enteric pathogens. In addition, the extended survival of what are generally referred to as stress-resistant pathogens suggests that gloves either be changed frequently during the day or washed in a disinfectant to reduce the risk of glove contamination that could otherwise contaminate product handled with the contaminated gloves. Copyright © 2017 Elsevier B.V. All rights reserved.

Prevalence and characterization of Escherichia coli and Salmonella strains isolated from stray dog and coyote feces in a major leafy greens production region at the United States-Mexico border.

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Michele T Jay-Russell

Full Text Available In 2010, Romaine lettuce grown in southern Arizona was implicated in a multi-state outbreak of Escherichia coli O145:H28 infections. This was the first known Shiga toxin-producing E. coli (STEC outbreak traced to the southwest desert leafy green vegetable production region along the United States-Mexico border. Limited information exists on sources of STEC and other enteric zoonotic pathogens in domestic and wild animals in this region. According to local vegetable growers, unleashed or stray domestic dogs and free-roaming coyotes are a significant problem due to intrusions into their crop fields. During the 2010-2011 leafy greens growing season, we conducted a prevalence survey of STEC and Salmonella presence in stray dog and coyote feces. Fresh fecal samples from impounded dogs and coyotes from lands near produce fields were collected and cultured using extended enrichment and serogroup-specific immunomagnetic separation (IMS followed by serotyping, pulsed-field gel electrophoresis (PFGE, and antimicrobial susceptibility testing. A total of 461 fecal samples were analyzed including 358 domestic dog and 103 coyote fecals. STEC was not detected, but atypical enteropathogenic E. coli (aEPEC strains comprising 14 different serotypes were isolated from 13 (3.6% dog and 5 (4.9% coyote samples. Salmonella was cultured from 33 (9.2% dog and 33 (32% coyote samples comprising 29 serovars with 58% from dogs belonging to Senftenberg or Typhimurium. PFGE analysis revealed 17 aEPEC and 27 Salmonella distinct pulsotypes. Four (22.2% of 18 aEPEC and 4 (6.1% of 66 Salmonella isolates were resistant to two or more antibiotic classes. Our findings suggest that stray dogs and coyotes in the desert southwest may not be significant sources of STEC, but are potential reservoirs of other pathogenic E. coli and Salmonella. These results underscore the importance of good agriculture practices relating to mitigation of microbial risks from animal fecal deposits in the

Antimicrobial Resistance of Shigella spp. isolated in the State of Pará, Brazil Resistência Antimicrobiana de Shigella spp. isoladas no Estado do Pará

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Flávia Corrêa Bastos

2011-10-01

Full Text Available INTRODUCTION: Shigella spp. are Gram-negative, nonsporulating, rod-shaped bacteria that belong to the family Enterobacteriaceae and are responsible for shigellosis or bacillary dysentery, an important cause of worldwide morbidity and mortality. METHODS: We studied the antibiotic resistance profiles of 122 Shigella spp. strains (81 S. flexneri, 41 S. sonnei, 1 S. boydii isolated from patients (female and male from 0 to 80 years of age presenting diarrhea in different districts of the State of Pará, in the North of Brazil. The antibiotic resistance of the strains, isolated from human fecal samples, was determined by the diffusion disk method and by using the VITEK-2 system. RESULTS: The highest resistance rate found was the resistance rate to tetracycline (93.8%, followed by the resistance rate to chloramphenicol (63.9% and to trimethoprim/sulfamethoxazole (63.1%. Resistance to at least three drugs was more common among S. flexneri than S. sonnei (39.5% vs. 10%. Six (4.9% strains were susceptible to all the antibiotics tested. All strains were susceptible to cefotaxime, ceftazidime, ciprofloxacin, nalidixic acid and nitrofurantoin. CONCLUSIONS: High rates of multidrug resistance in Shigella spp. are a serious public health concern in Brazil. It is extremely important to continuously monitor the antimicrobial resistances of Shigella spp. for effective therapy and control measures against shigellosis.INTRODUÇÃO: Shigella spp. são bactérias Gram-negativas, não esporuladas, em forma de bastonete, pertencentes a família Enterobacteriaceae responsáveis pela shigelose ou disenteria bacilar, uma importante causa de mortalidade e morbidade mundial. MÉTODOS: Foi estudado o perfil de resistência a antimicrobianos de 122 amostras de Shigella spp. (81 S. flexneri, 41 sonnei, 1 S. boydii isoladas de pacientes (sexo feminino e masculino com faixa etária de 0 a 80 anos com distúrbios gastrointestinais em diferentes municípios no Estado do Par

Survival of Salmonella, Escherichia coli O157:H7, and Listeria monocytogenes on raw peanut and pecan kernels stored at -24, 4, and 22°C.

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Brar, Pardeepinder K; Proano, Lisseth G; Friedrich, Loretta M; Harris, Linda J; Danyluk, Michelle D

2015-02-01

Cocktails of lawn-collected cells were used to determine the survival of Salmonella, Escherichia coli O157:H7, and Listeria monocytogenes on the surface of raw peanut and pecan kernels. Kernels were inoculated with mixtures of four to five strains at 3 or 6 log CFU/g, dried at room temperature, and then stored at -24 ± 1, 4 ± 2, and 22 ± 1°C for 28 or 365 days. In most cases, rates of decline of the pathogens did not differ significantly between the two inoculum concentrations in the 28-day study. At 6 log CFU/g, populations of all pathogens were reduced by 0.5 to 1.6 log CFU/g during an initial 3-day drying period on both peanuts and pecans. The moisture content of peanuts and pecans remained stable at -24 ± 1 and 22 ± 1°C; at 4 ± 2°C, the moisture content increased from 3.8 to 5.6% on peanuts and from 2.6 to 3% on pecans over 365 days. Pathogen populations were stable on pecans stored under frozen and refrigerated conditions, except for L. monocytogenes, which declined at a rate of 0.03 log CFU/g/30 days at 4 ± 2°C. Salmonella populations were stable on peanuts stored at -24 ± 1 and 4 ± 2°C, but E. coli O157:H7 and L. monocytogenes declined at rates of 0.03 to 0.12 log CFU/g/30 days. At 22 ± 1°C, Salmonella, E. coli O157:H7, and L. monocytogenes declined at a rate of 0.22, 0.37, and 0.59 log CFU/g/30 days, respectively, on peanuts, and at 0.15, 0.34, and 1.17 log CFU/g/30 days, respectively, on pecans. Salmonella counts were above the limit of detection (0.30 log CFU/g) throughout the study. In most cases during storage, counts obtained from pecans were higher than from peanuts.

Socioeconomic Determinants of Ciprofloxacin-Resistant Shigella Infections in Bangladeshi Children

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Randon J. Gruninger

2017-03-01

Full Text Available Background: Shigella species (spp. are a leading cause of moderate to severe diarrhea in children worldwide. The recent emergence of quinolone-resistant Shigella spp. gives cause for concern, and South Asia has been identified as a reservoir for global spread. The influence of socioeconomic status on antimicrobial resistance in developing countries, such as those in South Asia, remains unknown. Methods: We used data collected from 2009 to 2014 from a hospital specializing in the treatment of diarrhea in Dhaka, Bangladesh, to determine the relationship between ciprofloxacin-resistant Shigella spp. isolates and measures of socioeconomic status in Bangladeshi children less than 5 years of age. Results: We found 2.7% (230/8, 672 of children who presented with diarrhea had Shigella spp. isolated from their stool, and 50% (115/230 had resistance to ciprofloxacin. Using multivariable logistic regression analysis, we found that children from families where the father’s income was in the highest quintile had significantly higher odds of having ciprofloxacin-resistant Shigella spp. compared to children in the lowest quintile (OR = 6.1, CI 1.9-19. Factors protective against the development of resistance included access to improved sanitation (OR = 0.27, CI 0.11-0.7, and improved water sources (OR = 0.48, CI 0.25-0.92. We did not find a relationship between ciprofloxacin resistance and other proxies for socioeconomic status, including the presence of animals in the home, nutritional status, paternal education level, and the number of family members in the home. Conclusions: Although the associations between wealth and antimicrobial resistance are not fully understood, possible explanations include increased access and use of antibiotics, greater access to healthcare facilities and thus resistant pathogens, or greater consumption of commercially produced foods prepared with antibiotics.

Pathogenesis of Shigella diarrhea: rabbit intestinal cell microvillus membrane binding site for Shigella toxin

International Nuclear Information System (INIS)

Fuchs, G.; Mobassaleh, M.; Donohue-Rolfe, A.; Montgomery, R.K.; Grand, R.J.; Keusch, G.T.

1986-01-01

This study examined the binding of purified 125 I-labeled shigella toxin to rabbit jejunal microvillus membranes (MVMs). Toxin binding was concentration dependent, saturable, reversible, and specifically inhibited by unlabeled toxin. The calculated number of toxin molecules bound at 4 0 C was 7.9 X 10(10) (3 X 10(10) to 2 X 10(11))/micrograms of MVM protein or 1.2 X 10(6) per enterocyte. Scatchard analysis showed the binding site to be of a single class with an equilibrium association constant, K, of 4.7 X 10(9) M-1 at 4 0 C. Binding was inversely related to the temperature of incubation. A total of 80% of the labeled toxin binding at 4 0 C dissociated from MVM when the temperature was raised to 37 0 C, but reassociated when the temperature was again brought to 4 0 C. There was no structural or functional change of MVM due to toxin as monitored by electron microscopy or assay of MVM sucrase activity. These studies demonstrate a specific binding site for shigella toxin on rabbit MVMs. The physiological relevance of this receptor remains to be determined

A comparison of self-reported joint symptoms following infection with different enteric pathogens: effect of HLA-B27

DEFF Research Database (Denmark)

Schiellerup, P.; Krogfelt, K.A.; Locht, H.

2008-01-01

with positive fecal culture for Salmonella, Campylobacter, Yersinia, Shigella, and E. coli were addressed by questionnaires inquiring about gastrointestinal (GI) symptoms and the occurrence of joint pain in a previously healthy joint within 4 weeks after onset of infection. A blood sample was requested for HLA......-B27 typing. RESULTS: Of 3146 patients invited, 2105 (67%) responded to the survey questionnaire. The triggering infections were Campylobacter, 1003; Salmonella, 619; E. coli, 290; Shigella, 102; and Yersinia, 91. JPrea was reported by 294 subjects: Campylobacter, 131 (13.1%); Salmonella, 104 (16.......8%); Yersinia, 21 (23.1%); Shigella, 10 (9.8%); and E. coli, 28 (9.7%). There was a significant association between severity of gastroenteritis and development of arthralgia (p = 0.001). The odds ratio (OR) for JPrea in an HLA-B27-positive individual was 2.62 (95% CI 1.67-3.93) for the entire group...

Immunization with r-Lactococcus lactis expressing outer membrane protein A of Shigella dysenteriae type-1: evaluation of oral and intranasal route of administration.

Science.gov (United States)

Yagnik, B; Sharma, D; Padh, H; Desai, P

2017-02-01

To evaluate the comparative immunogenic potential of food grade Lactococcus lactis expressing outer membrane protein A (OmpA) of Shigella dysenteriae type-1 (SD-1) when administered either orally or intranasally. OmpA of SD-1 was cloned and expressed first in Escherichia coli and then in L. lactis. Presence of recombinant gene was confirmed by restriction enzyme digestion and immunoblot analysis. Using immobilized metal affinity chromatography, OmpA was purified from recombinant E. coliBL21 (DE3) and subcutaneously administered in BALB/c mice. Detection of OmpA-specific IgG antibodies by enzyme-linked immunosorbent assay (ELISA) confirmed the immunogenicity of OmpA. In order to establish r-L. lactis as a mucosal delivery vehicle, it was administered orally and nasally in BALB/c mice. Serum IgG and faecal IgA were assessed through ELISA to compare the relative potential of immunization routes and immunogenic potential of r-L. lactis. Immunization via the oral route proved superior to intranasal exposure. Recombinant L. lactis expressing OmpA of SD-1 was found to be immunogenic. Oral administration of r-L. lactis elicited higher systemic and mucosal immune response when compared with the nasal route. Using food grade recombinant L. lactis has implications in the development of a prophylactic against multidrug-resistant Shigella, which can be used as a prospective vaccine candidate. Evaluating mucosal routes of immunization demonstrated that the oral route of administration elicited better immune response against OmpA of Shigella. © 2016 The Society for Applied Microbiology.

Attenuated Salmonella enterica serovar Typhi and Shigella flexneri 2a strains mucosally deliver DNA vaccines encoding measles virus hemagglutinin, inducing specific immune responses and protection in cotton rats.

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Pasetti, Marcela F; Barry, Eileen M; Losonsky, Genevieve; Singh, Mahender; Medina-Moreno, Sandra M; Polo, John M; Ulmer, Jeffrey; Robinson, Harriet; Sztein, Marcelo B; Levine, Myron M

2003-05-01

Measles remains a leading cause of child mortality in developing countries. Residual maternal measles antibodies and immunologic immaturity dampen immunogenicity of the current vaccine in young infants. Because cotton rat respiratory tract is susceptible to measles virus (MV) replication after intranasal (i.n.) challenge, this model can be used to assess the efficacy of MV vaccines. Pursuing a new measles vaccine strategy that might be effective in young infants, we used attenuated Salmonella enterica serovar Typhi CVD 908-htrA and Shigella flexneri 2a CVD 1208 vaccines to deliver mucosally to cotton rats eukaryotic expression plasmid pGA3-mH and Sindbis virus-based DNA replicon pMSIN-H encoding MV hemagglutinin (H). The initial i.n. dose-response with bacterial vectors alone identified a well-tolerated dosage (1 x 10(9) to 7 x 10(9) CFU) and a volume (20 micro l) that elicited strong antivector immune responses. Animals immunized i.n. on days 0, 28, and 76 with bacterial vectors carrying DNA plasmids encoding MV H or immunized parenterally with these naked DNA vaccine plasmids developed MV plaque reduction neutralizing antibodies and proliferative responses against MV antigens. In a subsequent experiment of identical design, cotton rats were challenged with wild-type MV 1 month after the third dose of vaccine or placebo. MV titers were significantly reduced in lung tissue of animals immunized with MV DNA vaccines delivered either via bacterial live vectors or parenterally. Since attenuated serovar Typhi and S. flexneri can deliver measles DNA vaccines mucosally in cotton rats, inducing measles immune responses (including neutralizing antibodies) and protection, boosting strategies can now be evaluated in animals primed with MV DNA vaccines.

Brote causado por Escherichia coli en Chalco, México

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Cortés-Ortiz Iliana Alejandra

2002-01-01

Full Text Available Objetivo. Identificar el agente causal del brote de diarrea asociado con el desbordamiento del canal de aguas negras en Chalco. Material y métodos. Estudio retrospectivo y transversal, efectuado en el Instituto de Diagnóstico y Referencia Epidemiológicos (InDRE, de la Secretaría de Salud, con 1 550 hisopos rectales para el aislamiento e identificación bioquímica de V. cholerae y enterobacterias, obtenidos de la población del Valle de Chalco, que presentó diarrea y vómito durante el desastre natural acontecido el 31 de mayo de 2000. El análisis de los resultados se efectuó por la diferencia entre las proporciones de dos poblaciones (prueba de Ji cuadrada. Las cepas de E. coli se hibridaron por "colony blot" para los grupos ETEC, EIEC, EPEC y EHEC. Resultados. El 0.45% correspondió a Salmonella: S. agona, S. infantis, S. enteritidis, S. muenchen, S. typhimurium; 0.06% a Shigella flexneri 3a, y 76.6% a E. coli: 62.2% a ETEC (44.6 % con LT, 11.2% con ST, y 44.1% con ambas sondas, 0.84% a EIEC (sonda ial, 0.84% a EPEC (sonda bundle-forming pilus BFP, 0.08% a E. coli enterohemorrágica no-O157:H7 (sonda pCVD419, y 36.02% no hibridó. No se encontró asociación entre E. coli patógena con la edad y género. Conclusiones. Escherichia coli podría ser responsable del brote de diarrea. Es importante conocer el agente etiológico del brote para encaminar las estrategias en el estudio y control sanitario del mismo.

Fractionation and purification of DNA methylases of Shigella sonnei

International Nuclear Information System (INIS)

Suchkov, S.V.; Nikol'skaya, I.I.; Debov, S.S.

1986-01-01

A comparative study was made of the possibilities of various types of column chromatography and isoelectrofocusing for the analysis of the set of DNA methylases of the strains Shigella sonnei 47. On the basic of cation-exchange chromatography, a simple, rapid, and convenient method has been developed for the production of a highly active summary preparations of DNA methylases. It has been shown that affinity chromatography on heparin-Sepharose provides for the separation of methylases according to the characteristic of specificity for the nitrogen base. The presence of six individuals enzymes of DNA methylation, differing in value of the isoelectric point, was demonstrated in Shigella sonnei 47 cells by the IEF* method. Increased ability of the DNA methylases of Shigella sonnei 47 for nonspecific aggregation during the process of fractionation was demonstrated, which makes the use of column chromatography unsuitable for the isolation and purification of individual methylating enzymes of the investigated strain. The advantages of the IEF method, as well as its potentialities from the standpoint of studying various molecular forms of site-specific enzymes, are discussed

Determination of foodborne pathogenic bacteria by multiplex PCR-microchip capillary electrophoresis with genetic algorithm-support vector regression optimization.

Science.gov (United States)

Li, Yongxin; Li, Yuanqian; Zheng, Bo; Qu, Lingli; Li, Can

2009-06-08

A rapid and sensitive method based on microchip capillary electrophoresis with condition optimization of genetic algorithm-support vector regression (GA-SVR) was developed and applied to simultaneous analysis of multiplex PCR products of four foodborne pathogenic bacteria. Four pairs of oligonucleotide primers were designed to exclusively amplify the targeted gene of Vibrio parahemolyticus, Salmonella, Escherichia coli (E. coli) O157:H7, Shigella and the quadruplex PCR parameters were optimized. At the same time, GA-SVR was employed to optimize the separation conditions of DNA fragments in microchip capillary electrophoresis. The proposed method was applied to simultaneously detect the multiplex PCR products of four foodborne pathogenic bacteria under the optimal conditions within 8 min. The levels of detection were as low as 1.2 x 10(2) CFU mL(-1) of Vibrio parahemolyticus, 2.9 x 10(2) CFU mL(-1) of Salmonella, 8.7 x 10(1) CFU mL(-1) of E. coli O157:H7 and 5.2 x 10(1) CFU mL(-1) of Shigella, respectively. The relative standard deviation of migration time was in the range of 0.74-2.09%. The results demonstrated that the good resolution and less analytical time were achieved due to the application of the multivariate strategy. This study offers an efficient alternative to routine foodborne pathogenic bacteria detection in a fast, reliable, and sensitive way.

Bacterial pathogen spectrum of acute diarrheal outpatients in an urbanized rural district in Southwest China

Directory of Open Access Journals (Sweden)

Yongming Zhou

2018-05-01

Full Text Available Objectives: To conduct a one-year pathogen surveillance of acute diarrheal disease based on outpatient clinics in township hospitals in rural Hongta District of Yunnan Province, China. Methods: Fecal specimens of acute diarrhea cases and relevant epidemiological information were collected. Salmonella, Shigella, Vibrio, Aeromonas, Plesiomonas shigelloides and diarrheogenic Escherichia coli (DEC were examined. Results: Among the 797 stool specimens sampled, 198 samples (24.8% were positive in pathogen isolation, and 223 strains were isolated. The order of isolation rates from high to low were DEC, Aeromonas, P. shigelloides, Salmonella, Shigella and Vibrio. The overall positive rate in middle school students and preschool children was relatively high; while the overall positive rate of less than 1-year-old infants and above 55 years olds was relatively low. The isolates were analyzed by pulsed-field gel electrophoresis (PFGE. Some cases had the same or very close onset time, and the isolates had similar PFGE patterns, suggesting a possible outbreak once occurred but was not detected by the current infectious disease reporting system. Conclusions: Pathogen infection and transmission in rapidly urbanized rural areas is a serious issue. There is a great need for a more sensitive and accurate mode of monitoring, reporting and outbreak identification of diarrheal disease. Keywords: Diarrheal disease, Diarrheogenic pathogen, Molecular typing, Surveillance, Bacterial pathogen

Microbial quality and prevalence of Salmonella and Listeria in eggs

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Manijeh Mahdavi

2012-01-01

Full Text Available Aims: This study was undertaken to determine the microbial quality and the prevalence of Salmonella and Listeria in table eggs in Isfahan, Iran. Materials and Methods: A total of 525 samples were randomly collected from various shops in Isfahan, Iran. Microbial quality of eggs evaluated by coliform count and total bacterial viable counts. Also, detection of Listeria and Salmonella in egg contents and on eggs shells was performed. Results: The mean of total viable bacteria and coliform counts in the egg contents were 3.95 × 10 4 CFU/g and 4.94 × 10 3 CFU/g, respectively. Salmonella and Listeria were not found on the shell or content of eggs. Enterobacteriaceae families were found in 357 of 525 (68.28% and 276 of 525 (52.44% of egg shell and egg content samples, respectively. Moreover, Pseudomonas aeruginosa was isolated from 175 (33.41% and 144 (25.37% of egg shell and egg content, respectively. The isolated Enterobacteriaceae were included: Enterobacter aerogenes, Escherichia coli, Klebsiella pneumoniae, Buttiauxella agrestis, Cedecea lapagei, Cedecea davisae and Erwinia herbicola. Conclusion: The findings of the present study indicate although Salmonella and Listeria were not found in egg samples; however, there is an urgent need to improve the hygienic level of consumed eggs.

Isolasi karakterisasi dan pengelompokan strain Salmonella typhi asal Kabupaten Sumba Barat Daya Nusa Tenggara Timur berdasarkan sifat-sifat fenotip

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Charis Amarantini

2012-02-01

Full Text Available Typhoid fever is highly endemic in the South-West Sumba Regency, East Nusa Tenggara. The incidence rate of the diseases is high estimated at 725/100,000. It is an acute systemic infection caused by Salmonella typhi. The clinical symptoms of the disease are extremely diverse, starting from the mild form to severe ones with the most feared complication being perforation within the small intestine. Therefore, it is important to perform isolation, characterization, and grouping of S. typhi strains from the blood culture in order to determined definitely diagnosis and the different phenotypic characteristics in the community. Isolation was done in selective and differential media: BacT/ALERT FA culture media, Selenite Cystine Broth, Chromocult Coliform Agar, MacConkey Agar, andSalmonella Shigella Agar. The typical colony of Salmonella was confirmed on Triple Sugar Iron Agar, Urea agar, and L-Lysinedecarboxylation media. Phenotypic characteristics of all isolates were identified using API 20E and API 50CHE diagnostics. Based on biochemical characteristics the result showed that 18 strains obtained from different geographical origins were diverse. Four strains have similarity value 100% while the remained strains have similarity value 86.3-98.4%. All of the strains were categorized in the species of S. typhi.

Prevalence and characterization of human Shigella infections in Henan Province, China, in 2006

DEFF Research Database (Denmark)

Xia, S.; Xu, B.; Huang, L.

2011-01-01

In 2006, 3531 fecal samples were collected from patients with diarrhea in Henan Province, China. A total of 467 (13.2%) Shigella strains were isolated and serotyped. Seventy-one Shigella flexneri strains were characterized by MIC determination, pulsed-field gel electrophoresis (PFGE), and detection...

Identification of CsrC and Characterization of Its Role in Epithelial Cell Invasion in Salmonella enterica Serovar Typhimurium

OpenAIRE

Fortune, Doreen R.; Suyemoto, Mitsu; Altier, Craig

2006-01-01

The csr regulatory system of Salmonella regulates the expression of the genes of Salmonella pathogenicity island 1 (SPI1) required for the invasion of epithelial cells. This system consists of the posttranscriptional regulator CsrA and an untranslated regulatory RNA, CsrB, that opposes the action of CsrA. Here we identify and characterize the role of a second regulatory RNA, CsrC, whose ortholog was discovered previously in Escherichia coli. We show that a mutant of csrC has only mild defects...

Mitochondria mediate septin cage assembly to promote autophagy of Shigella.

Science.gov (United States)

Sirianni, Andrea; Krokowski, Sina; Lobato-Márquez, Damián; Buranyi, Stephen; Pfanzelter, Julia; Galea, Dieter; Willis, Alexandra; Culley, Siân; Henriques, Ricardo; Larrouy-Maumus, Gerald; Hollinshead, Michael; Sancho-Shimizu, Vanessa; Way, Michael; Mostowy, Serge

2016-07-01

Septins, cytoskeletal proteins with well-characterised roles in cytokinesis, form cage-like structures around cytosolic Shigella flexneri and promote their targeting to autophagosomes. However, the processes underlying septin cage assembly, and whether they influence S. flexneri proliferation, remain to be established. Using single-cell analysis, we show that the septin cages inhibit S. flexneri proliferation. To study mechanisms of septin cage assembly, we used proteomics and found mitochondrial proteins associate with septins in S. flexneri-infected cells. Strikingly, mitochondria associated with S. flexneri promote septin assembly into cages that entrap bacteria for autophagy. We demonstrate that the cytosolic GTPase dynamin-related protein 1 (Drp1) interacts with septins to enhance mitochondrial fission. To avoid autophagy, actin-polymerising Shigella fragment mitochondria to escape from septin caging. Our results demonstrate a role for mitochondria in anti-Shigella autophagy and uncover a fundamental link between septin assembly and mitochondria. © 2016 The Authors. Published under the terms of the CC BY 4.0 license.

Rapid detection of food-borne Salmonella contamination using IMBs-qPCR method based on pagC gene

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Jiashun Wang

Full Text Available Abstract Detection of Salmonella is very important to minimize the food safety risk. In this study, the recombinant PagC protein and PagC antibody were prepared and coupled with immunomagnetic beads (IMBs to capture Salmonella cells from pork and milk samples. And then the SYBR Green qualitative PCR was developed to detect the pathogenic Salmonella. The results showed that the PagC polyclonal antiserum is of good specificity and the capture rate of 0.1 mg IMBs for Salmonella tended to be stable at the range of 70-74% corresponding to the concentrations between 101 and 104 CFU/mL. The method developed demonstrated high specificity for the positive Salmonella samples when compared to non-specific DNA samples, such as Escherichia coli, Staphylococcus aureus, Yersinia enterocolitica, and Yersinia pseudotuberculosis. The limit of detection of this assay was 18 CFU/mL. Detection and quantitative enumeration of Salmonella in samples of pork or milk shows good recoveries of 54.34% and 52.07%. In conclusion, the polyclonal antibody of recombinant PagC protein is effective to capture Salmonella from detected samples. The developed pagC antibody IMBs-qPCR method showed efficiency, sensitivity and specificity for 30 Salmonella detection, enabling detection within 10 h, which is a promising rapid method to detect Salmonella in emergency.

Association Between Shigella Infection and Diarrhea Varies Based on Location and Age of Children.

Science.gov (United States)

Lindsay, Brianna; Saha, Debasish; Sanogo, Doh; Das, Sumon Kumar; Omore, Richard; Farag, Tamer H; Nasrin, Dilruba; Li, Shan; Panchalingam, Sandra; Levine, Myron M; Kotloff, Karen; Nataro, James P; Magder, Laurence; Hungerford, Laura; Faruque, A S G; Oundo, Joseph; Hossain, M Anowar; Adeyemi, Mitchell; Stine, Oscar Colin

2015-11-01

Molecular identification of the invasion plasmid antigen-H (ipaH) gene has been established as a useful detection mechanism for Shigella spp. The Global Enteric Multicenter Study (GEMS) identified the etiology and burden of moderate-to-severe diarrhea (MSD) in sub-Saharan Africa and south Asia using a case-control study and traditional culture techniques. Here, we used quantitative polymerase chain reaction (qPCR) to identify Shigella spp. in 2,611 stool specimens from GEMS and compared these results to those using culture. Demographic and nutritional characteristics were assessed as possible risk factors. The qPCR identified more cases of shigellosis than culture; however, the distribution of demographic characteristics was similar by both methods. In regression models adjusting for Shigella quantity, age, and site, children who were exclusively breast-fed had significantly lower odds of MSD compared with children who were not breast-fed (odds ratio [OR] = 0.47, 95% confidence interval (CI) = 0.28-0.81). The association between Shigella quantity and MSD increased with age, with a peak in children of 24-35 months of age (OR = 8.2, 95% CI = 4.3-15.7) and the relationship between Shigella quantity and disease was greatest in Bangladesh (OR = 13.2, 95% CI = 7.3-23.8). This study found that qPCR identified more cases of Shigella and age, site, and breast-feeding status were significant risk factors for MSD. © The American Society of Tropical Medicine and Hygiene.

Inactivation of Nonpathogenic Escherichia coli, Escherichia coli O157:H7, Salmonella enterica Typhimurium, and Listeria monocytogenes in Ice Using a UVC Light-Emitting Diode.

Science.gov (United States)

Murashita, Suguru; Kawamura, Shuso; Koseki, Shigenobu

2017-07-01

Ice, widely used in the food industry, is a potential cause of food poisoning resulting from microbial contamination. Direct microbial inactivation of ice is necessary because microorganisms may have been present in the source water used to make it and/or may have been introduced due to poor hygiene during production or handling of the ice. Nonthermal and nondestructive microbial inactivation technologies are needed to control microorganisms in ice. We evaluated the applicability of a UVC light-emitting diode (UVC-LED) for microbial inactivation in ice. The effects of UV intensity and UV dose of the UVC-LED on Escherichia coli ATCC 25922 and a comparison of UVC-LED with a conventional UV lamp for effective bacterial inactivation in distilled water and ice cubes were investigated to evaluate the performance of the UVC-LED. Finally, we assessed the effects of the UVC-LED on pathogens such as E. coli O157:H7, Salmonella Typhimurium, and Listeria monocytogenes in ice cubes. The results indicated that UVC-LED effectiveness depended on the UV dose at all UV intensity conditions (0.084, 0.025, 0.013, 0.007, and 0.005 mW/cm 2 ) in ice and that UVC-LED could more efficiently inactivate E. coli ATCC 25922 in distilled water and ice than the UV lamp. At a UV dose of 2.64 mJ/cm 2 , E. coli in distilled water was decreased by 0.90 log CFU/mL (UV lamp) and by more than 7.0 log CFU/mL (UVC-LED). At 15.2 mJ/cm 2 , E. coli in ice was decreased by 3.18 log CFU/mL (UV lamp) and by 4.45 CFU/mL (UVC-LED). Furthermore, UVC-LED irradiation reduced the viable number of pathogens by 6 to 7 log cycles at 160 mJ/cm 2 , although the bactericidal effect was somewhat dependent on the type of bacteria. L. monocytogenes in ice was relatively more sensitive to UVC irradiation than were E. coli O157:H7 and Salmonella Typhimurium. These results demonstrate that UVC-LED irradiation could contribute to the safety of ice in the food industry.

Cloning of regions required for contact hemolysis and entry into LLC-MK2 cells from Shigella sonnei form I plasmid: virF is a positive regulator gene for these phenotypes.

OpenAIRE

Kato, J; Ito, K; Nakamura, A; Watanabe, H

1989-01-01

Two distinct regions required for both contact hemolysis and entry into LLC-MK2 cells were cloned into Escherichia coli from the Shigella sonnei form I plasmid, pSS120. The first region was cloned into an E. coli HB101 strain containing noninvasive Tn1 insertion mutants of the form I plasmid, and expression of ipa (invasion plasmid antigen) gene products was restored. The plasmid carrying the first region was then transformed into E. coli lacking the form I plasmid, and additional DNA fragmen...

Antimicrobial resistance in Shigella spp. causing traveller's diarrhoea (1995-2010): a retrospective analysis.

Science.gov (United States)

Pons, M J; Gomes, C; Martínez-Puchol, S; Ruiz, L; Mensa, L; Vila, J; Gascón, J; Ruiz, J

2013-01-01

Shigellosis is a global human health problem causing an important morbidity among travellers returning from tropical areas. This study was aimed to describe the evolution of antimicrobial resistance profile in Shigella spp. isolated between the years 1995-2010 in patients with traveller's diarrhoea (TD) returning from tropical areas. The levels of antimicrobial resistance were tested in a total of 191 Shigella spp. isolated during the period from 1995 to 2010. A decrease of cases of diarrhoea caused by Shigella has been observed in recent years. A wide spectrum of antibiotic resistance was observed among Shigella spp. These isolates showed high levels of resistance to tetracycline (84%), co-trimoxazole (75.5%), and ampicillin (45.5%). The resistance was low to ciprofloxacin (2.1%), azithromycin (3.9%) and furazolidone (8.4%). According to the period, in the case of ampicillin, amoxicillin plus clavulanic acid, chloramphenicol, values of resistance were significantly decreasing from 1995-2000 to 2001-2010, (62.5% vs. 28.4%, 19.8% vs. 6.6%, 23.4 vs. 10.4%, respectively). Meanwhile in nalidixic acid and tetracycline the evolution of resistance has increased over time. A decrease in the isolation number of Shigella spp. causing TD has been observed. Differential trends in the evolution of the levels of resistance to the tested antibacterial agents have been observed. Copyright © 2013 Elsevier Ltd. All rights reserved.

Prevalence of Shigella among diarrheic children under-5 years of age attending at Mekelle health center, north Ethiopia.

Science.gov (United States)

Kahsay, Atsebaha Gebrekidan; Teklemariam, Zelalem

2015-12-15

Shigellosis is recognized as a major global public health problem especially in developing countries particularly in children under-5 years of age. Therefore; the objective of this study was to determine the prevalence of Shigella among diarrheic children under-5 years of age attending at Mekelle health center, north Ethiopia. A cross-sectional study was conducted among diarrheic children under-5 years of age from March to May, 2012. Structured questionnaire was used to collect the data. Study participants were recruited by convenience sampling technique. Shigella was isolated and identified using standard bacteriological techniques. A total of 241 study participants were included in the study. The overall prevalence of Shigella in this study was 13.3% (32/241). High prevalence of Shigella (22.6%) was revealed from the age group of 12-23 months. No Shigella was isolated from the age group of 0-5 months. Majority of the isolates of Shigella were from bloody and mucoid diarrhea. There was high prevalence of Shigella infection in this study. Children among the age group of 12-23 months were highly affected. Therefore; responsible bodies should work hard on preventive measures to reduce or eradicate the problem occurred due to shigellosis.

Enteric Infections Circulating during Hajj Seasons, 2011–2013

KAUST Repository

Abd El Ghany, Moataz; Alsomali, Mona; Almasri, Malak; Padron Regalado, Eriko; Naeem, Raeece; Tukestani, AbdulHafeez; Asiri, Abdullah; Hill-Cawthorne, Grant A.; Pain, Arnab; Memish, Ziad A.

2017-01-01

Hajj, the annual Muslim pilgrimage to Mecca, Saudi Arabia, is a unique mass gathering event that raises public health concerns in the host country and globally. Although gastroenteritis and diarrhea are common among Hajj pilgrims, the microbial etiologies of these infections are unknown. We collected 544 fecal samples from pilgrims with medically attended diarrheal illness from 40 countries during the 2011-2013 Hajj seasons and screened the samples for 16 pathogens commonly associated with diarrheal infections. Bacteria were the main agents detected, in 82.9% of the 228 positive samples, followed by viral (6.1%) and parasitic (5.3%) agents. Salmonella spp., Shigella/enteroinvasive Escherichia coli, and enterotoxigenic E. coli were the main pathogens associated with severe symptoms. We identified genes associated with resistance to third-generation cephalosporins approximate to 40% of Salmonella- and E. coli-positive samples. Hajj-associated foodborne infections pose a major public health risk through the emergence and transmission of antimicrobial drug-resistant bacteria.

Enteric Infections Circulating during Hajj Seasons, 2011–2013

KAUST Repository

Abd El Ghany, Moataz

2017-09-13

Hajj, the annual Muslim pilgrimage to Mecca, Saudi Arabia, is a unique mass gathering event that raises public health concerns in the host country and globally. Although gastroenteritis and diarrhea are common among Hajj pilgrims, the microbial etiologies of these infections are unknown. We collected 544 fecal samples from pilgrims with medically attended diarrheal illness from 40 countries during the 2011-2013 Hajj seasons and screened the samples for 16 pathogens commonly associated with diarrheal infections. Bacteria were the main agents detected, in 82.9% of the 228 positive samples, followed by viral (6.1%) and parasitic (5.3%) agents. Salmonella spp., Shigella/enteroinvasive Escherichia coli, and enterotoxigenic E. coli were the main pathogens associated with severe symptoms. We identified genes associated with resistance to third-generation cephalosporins approximate to 40% of Salmonella- and E. coli-positive samples. Hajj-associated foodborne infections pose a major public health risk through the emergence and transmission of antimicrobial drug-resistant bacteria.

Safety and Immunogenicity of a Shigella flexneri 2a Invaplex 50 Intranasal Vaccine in Adult Volunteers

Science.gov (United States)

2010-06-01

Scandjlnfect Dis 1993;25(5):569-77. [11] Oaks EV, Hale TL. Formal SB. Serum immune response to Shigella protein ami- gens in rhesus monkeys and humans...infection with virulent Shigellaflexneri 2a on the resistance of monkeys to subsequent infection with Shigella sonnei. J Infect Dis 1991; 164(September...july 2010 Keywords: Shigella jlexneri lnvaplex Nasal vaccine Shigellosis is a leading cause of diarrhea worldwide prompting vaccine development

Salmonella infections modelling in Mississippi using neural network and geographical information system (GIS).

Science.gov (United States)

Akil, Luma; Ahmad, H Anwar

2016-03-03

Mississippi (MS) is one of the southern states with high rates of foodborne infections. The objectives of this paper are to determine the extent of Salmonella and Escherichia coli infections in MS, and determine the Salmonella infections correlation with socioeconomic status using geographical information system (GIS) and neural network models. In this study, the relevant updated data of foodborne illness for southern states, from 2002 to 2011, were collected and used in the GIS and neural networks models. Data were collected from the Centers for Disease Control and Prevention (CDC), MS state Department of Health and the other states department of health. The correlation between low socioeconomic status and Salmonella infections were determined using models created by several software packages, including SAS, ArcGIS @RISK and NeuroShell. Results of this study showed a significant increase in Salmonella outbreaks in MS during the study period, with highest rates in 2011 (47.84 ± 24.41 cases/100,000; pGIS maps of Salmonella outbreaks in MS in 2010 and 2011 showed the districts with higher rates of Salmonella. Regression analysis and neural network models showed a moderate correlation between cases of Salmonella infections and low socioeconomic factors. Poverty was shown to have a negative correlation with Salmonella outbreaks (R(2)=0.152, p<0.05). Geographic location besides socioeconomic status may contribute to the high rates of Salmonella outbreaks in MS. Understanding the geographical and economic relationship with infectious diseases will help to determine effective methods to reduce outbreaks within low socioeconomic status communities. Published by the BMJ Publishing Group Limited. For permission to use (where not already granted under a licence) please go to http://www.bmj.com/company/products-services/rights-and-licensing/

The Galleria mellonella larvae as an in vivo model for evaluation of Shigella virulence.

Science.gov (United States)

Barnoy, Shoshana; Gancz, Hanan; Zhu, Yuewei; Honnold, Cary L; Zurawski, Daniel V; Venkatesan, Malabi M

2017-07-04

Shigella spp. causing bacterial diarrhea and dysentery are human enteroinvasive bacterial pathogens that are orally transmitted through contaminated food and water and cause bacillary dysentery. Although natural Shigella infections are restricted to humans and primates, several smaller animal models are used to analyze individual steps in pathogenesis. No animal model fully duplicates the human response and sustaining the models requires expensive animals, costly maintenance of animal facilities, veterinary services and approved animal protocols. This study proposes the development of the caterpillar larvae of Galleria mellonella as a simple, inexpensive, informative, and rapid in-vivo model for evaluating virulence and the interaction of Shigella with cells of the insect innate immunity. Virulent Shigella injected through the forelegs causes larvae death. The mortality rates were dependent on the Shigella strain, the infectious dose, and the presence of the virulence plasmid. Wild-type S. flexneri 2a, persisted and replicated within the larvae, resulting in haemocyte cell death, whereas plasmid-cured mutants were rapidly cleared. Histology of the infected larvae in conjunction with fluorescence, immunofluorescence, and transmission electron microscopy indicate that S. flexneri reside within a vacuole of the insect haemocytes that ultrastructurally resembles vacuoles described in studies with mouse and human macrophage cell lines. Some of these bacteria-laden vacuoles had double-membranes characteristic of autophagosomes. These results suggest that G. mellonella larvae can be used as an easy-to-use animal model to understand Shigella pathogenesis that requires none of the time and labor-consuming procedures typical of other systems.

Detection and Characterization of Shiga Toxin Producing Escherichia coli, Salmonella spp., and Yersinia Strains from Human, Animal, and Food Samples in San Luis, Argentina

Science.gov (United States)

Favier, Gabriela Isabel; Lucero Estrada, Cecilia; Cortiñas, Teresa Inés; Escudero, María Esther

2014-01-01

Shiga toxin producing Escherichia coli (STEC), Salmonella spp., and Yersinia species was investigated in humans, animals, and foods in San Luis, Argentina. A total of 453 samples were analyzed by culture and PCR. The antimicrobial susceptibility of all the strains was studied, the genomic relationships among isolates of the same species were determined by PFGE, and the potencial virulence of Y. enterocolitica strains was analyzed. Yersinia species showed higher prevalence (9/453, 2.0%, 95% CI, 0.7–3.3%) than STEC (4/453, 0.9%, 95% CI, 0–1.8%) and Salmonella spp. (3/453, 0.7%, 95% CI, 0–1.5%). Y. enterocolitica and Y. intermedia were isolated from chicken carcasses (6/80, 7.5%, 95% CI, 1.5–13.5%) and porcine skin and bones (3/10, 30%, 95% CI, 0–65%). One STEC strain was recovered from human feces (1/70, 1.4%, 95% CI, 0–4.2%) and STEC stx1/stx2 genes were detected in bovine stools (3/129, 2.3%, 95% CI, 0–5.0%). S. Typhimurium was isolated from human feces (1/70, 1.4%, 95% CI, 0–4.2%) while one S. Newport and two S. Gaminara strains were recovered from one wild boar (1/3, 33%, 95% CI, 0–99%). The knowledge of prevalence and characteristics of these enteropathogens in our region would allow public health services to take adequate preventive measures. PMID:25177351

Force Protection Technologies for the 2010-2020 Timeframe

Science.gov (United States)

2003-11-01

V11. Marburg virus V12. Monkey pox virus Viruses V13. Rift Valley fever virus V14. Tick-borne encephalitis virus (Russian Spring-Summer...pseudomallei (pseudomonas pseudomallei) B10. Salmonella typhi B11. Shigella dysenteriae Bacteria B11. Vibrio cholerae Sensor Section, ASB...and shown to detect Shigella flexneri and Shigella dysenteriae genes. Each class of threat agent is likely to give a distinct hybridization pattern

Antimicrobial resistance in Escherichia coli and Salmonella spp. isolates from fresh produce and the impact to food safety.

Science.gov (United States)

Vital, Pierangeli G; Caballes, Marie Bernadine D; Rivera, Windell L

2017-09-02

Foodborne diseases associated with fresh produce consumption have escalated worldwide, causing microbial safety of produce of critical importance. Bacteria that have increasingly been detected in fresh produce are Escherichia coli and Salmonella spp., both of which have been shown to progressively display antimicrobial resistance. The study focused on the assessment of antimicrobial resistance of these enteric bacteria from different kinds of fresh produce from various open air markets and supermarkets in the Philippines. Using the disk diffusion assay on a total of 50 bacterial isolates obtained from 410 fresh produce surveyed, monoresistance to tetracycline was observed to be the most prevalent (38%), followed by multidrug resistance to tetracycline, chloramphenicol, ciprofloxacin, and nalidixic acid (4%), and lastly by dual resistance to tetracycline and chloramphenicol (2%). Using multiplex and simplex polymerase chain reaction (PCR) assays, tetA (75%) and tetB (9%) were found in tetracycline resistant isolates, whereas catI (67%) and catIII (33%) were detected in chloramphenicol resistant isolates. Sequence analysis of gyr and par genes from the ciprofloxacin and nalidixic acid resistant isolates revealed different mutations. Based on the results, fresh produce act as a reservoir of these antibiotic resistant bacteria which may pose health threat to consumers.

Synthesis of new derivatives of hydrazinecarbothioamides and 1,2,4-triazoles and evaluation of their antimicrobial activities

Directory of Open Access Journals (Sweden)

Socea Laura I.

2015-01-01

Full Text Available A new series of hydrazinecarbothioamides 6-9 bearing 5H-dibenzo[a,d][7]annulene moiety were synthesized. Cyclization of 6-9 in NaOH solution produced the corresponding 1,2,4-triazoles-3(4H-thiol 10-13, which proved to be axial isomers. The thioethers 14-17 were prepared by alkylation of 10-13 with methyl iodide. All new compounds were characterized by elemental analysis, IR-, UV-, 1H-NMR and 13C-NMR spectroscopy. The evaluation for antimicrobial activity against Staphylococcus aureus ATCC 25923, Pseudomonas aeruginosa ATCC 27853, Escherichia coli ATCC 25922, Bacillus subtilis ATCC 6663, Salmonella tiphimurium ATCC 14028, Shigella flexneri ATCC 12022, Candida albicans ATCC 90028 was performed.

Triazenos e atividade antibacteriana Triazenes and antibacterial activity

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