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Sample records for enterica serotype agona

  1. A recurrent, multistate outbreak of salmonella serotype agona infections associated with dry, unsweetened cereal consumption, United States, 2008.

    PubMed

    Russo, Elizabeth T; Biggerstaff, Gwen; Hoekstra, R Michael; Meyer, Stephanie; Patel, Nehal; Miller, Benjamin; Quick, Rob

    2013-02-01

    An outbreak of Salmonella enterica serotype Agona infections associated with nationwide distribution of cereal from Company X was identified in April 2008. This outbreak was detected using PulseNet, the national molecular subtyping network for foodborne disease surveillance, which coincided with Company X's voluntary recall of unsweetened puffed rice and wheat cereals after routine product sampling yielded Salmonella Agona. A case patient was defined as being infected with the outbreak strain of Salmonella Agona, with illness onset from 1 January through 1 July 2008. Case patients were interviewed using a standard questionnaire, and the proportion of ill persons who reported eating Company X puffed rice cereal was compared with Company X's market share data using binomial testing. The Minnesota Department of Agriculture inspected the cereal production facility and collected both product and environmental swab samples. Routine surveillance identified 33 case patients in 17 states. Of 32 patients interviewed, 24 (83%) reported eating Company X puffed rice cereal. Company X puffed rice cereal represented 0.063% of the total ready-to-eat dry cereal market share in the United States at the time of the investigation. Binomial testing suggested that the proportion of exposed case patients would not likely occur by chance (P < 0.0001). Of 17 cereal samples collected from case patient homes for laboratory testing, 2 (12%) yielded Salmonella Agona indistinguishable from the outbreak strain. Twelve environmental swabs and nine product samples from the cereal plant yielded the outbreak strain of Salmonella Agona. Company X cereal was implicated in a similar outbreak of Salmonella Agona infection in 1998 with the same outbreak strain linked to the same production facility. We hypothesize that a recent construction project at this facility created an open wall near the cereal production area allowing reintroduction of Salmonella Agona into the product, highlighting the

  2. Complete Genome Sequence of Salmonella enterica Serovar Agona Pulsed-Field Type SAGOXB.0066, Cause of a 2008 Pan-European Outbreak.

    PubMed

    McCusker, Matthew P; Hokamp, Karsten; Buckley, James F; Wall, Patrick G; Martins, Marta; Fanning, Séamus

    2014-01-01

    Salmonella enterica serovar Agona is in the top 10 most common nontyphoidal serovars reported in humans in the European Union. Here we report the complete genome sequence of an S. enterica serovar Agona isolate, designated 24249, that was the cause of a pan-European outbreak in 2008 with 163 confirmed cases reported. PMID:24459278

  3. Clinical Isolates of Salmonella enterica Serovar Agona Producing NDM-1 Metallo-β-Lactamase: First Report from Pakistan

    PubMed Central

    Khan, Erum; Jabeen, Kauser; Bhawan, Pushpa; Hopkins, Katie L.; Day, Martin; Nasir, Amna; Meunier, Daniele; Woodford, Neil

    2014-01-01

    We report two cases of infantile diarrhea due to multidrug-resistant, NDM-1 metallo-β-lactamase-producing Salmonella enterica serovar Agona from Pakistan. This study alerts toward possible risk of NDM-1 transmission to enteric fever pathogens and encourages microbiologists to consider active screening of carbapenem resistance in nontyphoidal Salmonella isolates. PMID:25378577

  4. Assignment of serotype to Salmonella enterica isolates obtained from poultry and their environment in southern Brazil

    PubMed Central

    Pulido-Landínez, M; Sánchez-Ingunza, R; Guard, J; do Nascimento, V Pinheiro

    2013-01-01

    To assess diversity of Salmonella enterica serotypes present in poultry and their environment from southern Brazil, the Kauffmann–White–Le Minor (KWL) scheme was used to serotype a total of 155 isolates. Isolates were then re-examined with nested PCR and sequencing of the dkgB-linked intergenic sequence ribotyping (ISR) region that assesses single nucleotide polymorphisms occurring around a 5S ribosomal gene. Serotypes identified were Heidelberg (40·6%), Enteritidis (34·2%), Hadar (8·4%), Typhimurium (3·9%), Gallinarum (3·2%), Agona (1·3%), Cerro (1·3%), Livingstone (1·3%), Infantis (0·6%), Isangi (0·6%), Mbandaka (0·6%), Montevideo (0·6%) and Senftenberg (0·6%). Three unique ISRs were detected from four strains. Day old chicks yielded only S. Enteritidis, whereas S. Heidelberg was most often associated with poultry carcasses. Overall agreement between KWL and ISR was 85·2%, with disagreement possibly due to the ability of ISR to detect mixtures of serotypes in culture. Overall, ISR provided more information than did KWL about the ecology of Salm. enterica on-farm. The O-antigen group D Salm. enterica serovars such as Pullorum, Gallinarum and Enteritidis appear susceptible to overgrowth by other serotypes. Significance and Impact of the Study Single nucleotide polymorphisms found in a group of poultry-associated Salmonella isolates from southern Brazil provided evidence of mixtures of serovar group D serotypes on-farm and in single samples from birds. This finding suggests that co-infection and interserotype competition of Salmonella enterica in poultry could impact the incidence of disease in animals or humans. In addition, unique serotypes were identified on-farm that escaped characterization by antibody typing. Application of cost-efficient and highly discriminatory genomic methods for assigning serotype may alter concepts about the epidemiology of Salm. enterica on-farm and in foods. PMID:23734786

  5. Detection of Plasmids and Class 1 Integrons in Salmonella enterica serovar Agona Isolated from NARMS Slaughter Samples Collected in Years 1997 through 2003

    Technology Transfer Automated Retrieval System (TEKTRAN)

    A total of 60 Salmonella enterica serovar Agona isolates (25 pan-susceptible isolates and 35 isolates resistant to five or more antimicrobials) submitted to the National Antimicrobial Resistance Monitoring System –Enteric Bacteria (NARMS) from 1997 through 2003 were examined for plasmids and class 1...

  6. Development of a rapid serotyping method for Salmonella enterica using serotype-specific single-nucleotide polymorphisms

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Salmonella enterica subsp. enterica serotype Enteriditis (S. Enteriditis) is the leading cause of salmonellosis worldwide, including the USA. Many S. enterica serotypes known to cause foodborne disease are associated with broiler meat contamination. While some serotypes are specific to birds (S. e...

  7. Draft Genome Sequence of Salmonella enterica subsp. enterica Serotype Saintpaul Strain S-70, Isolated from an Aquatic Environment

    PubMed Central

    Estrada-Acosta, Mitzi; Medrano-Félix, Andrés; Jiménez, Maribel; Gómez-Gil, Bruno; León-Félix, Josefina; Amarillas, Luis

    2013-01-01

    Salmonella is a pathogen of worldwide importance, causing disease in a vast range of hosts, including humans. We report the genome sequence of Salmonella enterica subsp. enterica serotype Saintpaul strain S-70, isolated from an aquatic environment. PMID:24336367

  8. Rapid molecular pathotyping of major salmonella enterica serotypes based on single-nucleotide polymorphisms (SNPs) in the adenylate cyclase (cyaA) gene

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Introduction: Salmonella enterica subsp. enterica serotype Enteriditis (S. Enteriditis) is the leading cause of salmonellosis worldwide, including the USA. Many S. enterica serotypes known to cause foodborne disease are associated with broiler meat contamination. While some serotypes are specific...

  9. A MULTIPLEX PCR METHOD FOR THE RAPID SEROTYPING OF COMMON CLINICAL ISOLATES OF SALMONELLA ENTERICA SUBSPECIES ENTERICA

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Background: The bacterial species Salmonella enterica is one of the major causes of gastroenteritis in humans and has over 1,500 serotypes. Serotyping is the most common tool used to identify isolates from diseased patients. However, the serotyping method can take several weeks and sometimes can ...

  10. Salmonella enterica Serotype Choleraesuis: Epidemiology, Pathogenesis, Clinical Disease, and Treatment†

    PubMed Central

    Chiu, Cheng-Hsun; Su, Lin-Hui; Chu, Chishih

    2004-01-01

    Nontyphoid Salmonella strains are important causes of reportable food-borne infection. Among more than 2,000 serotypes, Salmonella enterica serotype Choleraesuis shows the highest predilection to cause systemic infections in humans. The most feared complication of serotype Cholearesuis bacteremia in adults is the development of mycotic aneurysm, which previously was almost uniformally fatal. The advances in diagnostic techniques, surgical care, and antimicrobial therapy have greatly improved the survival of these patients. However, the recent emergence of serotype Choleraesuis that is resistant to ampicillin, chloramphenicol, trimethoprim-sulfamethoxazole, and, notably, fluoroquinolone antibiotics has aroused concern about the use of these agents for the empirical treatment of systemic infection caused by this organism. In view of the serious implications of the situation, the chain of transmission and mechanism of resistance should be carefully studied to reduce the spread of infection and threat to human health. To date, there are no vaccines available to prevent serotype Choleraesuis infections in humans. The availability, in the near future, of the genome sequence of serotype Cholearesuis will facilitate the development of effective vaccines as well as the discovery of new targets for novel antimicrobial agents. PMID:15084503

  11. Draft Genome Sequence of Salmonella enterica subsp. enterica Serotype Oranienburg Strain S-76, Isolated from an Aquatic Environment

    PubMed Central

    Medrano-Félix, Andrés; Estrada-Acosta, Mitzi; Jiménez, Maribel; Gómez-Gil, Bruno; León-Félix, Josefina; Amarillas, Luis

    2013-01-01

    Salmonella is a widespread microorganism and a common causative agent of food-borne illnesses. Salmonella enterica subsp. enterica serotype Oranienburg is highly prevalent in surface water from tropical ecosystems and is not commonly related to illnesses. Here, we report the first genome sequence of Salmonella Oranienburg strain S-76, isolated from an aquatic environment. PMID:24336368

  12. Development and application of novel SNP-based serotyping assays in targeting Salmonella enterica within the poultry production and processing continuum.

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Salmonella enterica subsp. enterica serotype Enteriditis (S. Enteriditis) is the leading cause of salmonellosis worldwide. While some S. enterica serotypes are specific to birds, many represent human zoonotic pathogens, thus their presence and survival throughout the continuum of poultry production...

  13. Assignment of serotype to Salmonella enterica isolates obtained from poultry and their environment in Southern Brazil.

    Technology Transfer Automated Retrieval System (TEKTRAN)

    To assess diversity of Salmonella enterica serotypes present in poultry and their environment from Southern Brazil, the Kauffman-White-LeMinor (KWL) scheme was used to serotype a total of 155 isolates. Isolates were then re-examined with nested PCR and sequencing of the dkgB-linked Intergenic Sequ...

  14. Development of a Rapid Multiplex PCR Technique for Determination of Salmonella enterica Serotypes Isolated from Pork and Poultry

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Background: A multiplex PCR technique to discriminate Salmonella enterica serotypes was adapted to a high-throughput, automated assay. Methods: Fifteen target genes were chosen that varied in distribution among common Salmonella enterica serotypes isolated from various hosts. These targets were dete...

  15. Genomic Epidemiology of Salmonella enterica Serotype Enteritidis based on Population Structure of Prevalent Lineages

    PubMed Central

    Desai, Prerak T.; den Bakker, Henk C.; Mikoleit, Matthew; Tolar, Beth; Trees, Eija; Hendriksen, Rene S.; Frye, Jonathan G.; Porwollik, Steffen; Weimer, Bart C.; Wiedmann, Martin; Weinstock, George M.; Fields, Patricia I.; McClelland, Michael

    2014-01-01

    Salmonella enterica serotype Enteritidis is one of the most commonly reported causes of human salmonellosis. Its low genetic diversity, measured by fingerprinting methods, has made subtyping a challenge. We used whole-genome sequencing to characterize 125 S. enterica Enteritidis and 3 S. enterica serotype Nitra strains. Single-nucleotide polymorphisms were filtered to identify 4,887 reliable loci that distinguished all isolates from each other. Our whole-genome single-nucleotide polymorphism typing approach was robust for S. enterica Enteritidis subtyping with combined data for different strains from 2 different sequencing platforms. Five major genetic lineages were recognized, which revealed possible patterns of geographic and epidemiologic distribution. Analyses on the population dynamics and evolutionary history estimated that major lineages emerged during the 17th–18th centuries and diversified during the 1920s and 1950s. PMID:25147968

  16. Real-time FRET PCR assay for Salmonella enterica serotype detection in food.

    PubMed

    Olsen, Eric V; Gibbins, Carl S; Grayson, J Kevin

    2009-09-01

    Salmonella enterica subsp. enterica serotypes are leading etiological agents of food-borne gastroenteritis. Traditional identification is laborious and time intensive. Faster molecular methods may allow early identification in contaminated food products. We developed a real-time, fluorescence resonance energy transfer hybridization probe polymerase chain reaction (PCR) assay for S. enterica serotypes on the basis of the exclusive presence of the apeE gene in Salmonella Typhimurium. Assay sensitivity for 12 S. enterica serotypes was as low as 1.87 x 10(2) genomic equivalents per milliliter. PCR efficiency was 94% and the dynamic range was linear over six orders of magnitude from 10(0) to 10(6) copies. The lower limit of detection for 12 different food matrices was between 1.5 x 10(2) and 1.5 x 10(5) CFU/mL without pre-enrichment. When combined with high-throughput automated DNA extraction, 32 food specimens were processed and assayed in less than 2 hours, allowing rapid, specific, sensitive detection of S. enterica serotypes in food products. PMID:19780376

  17. Salmonella enterica Serotype Choleraesuis Infection of the Knee and Femur in a Nonbacteremic Diabetic Patient

    PubMed Central

    Sy, Alexander M.; Sandhu, Jagbir; Lenox, Theodore

    2013-01-01

    Osteoarticular infections caused by Salmonella are rare. The rates of osteomyelitis and septic arthritis due to Salmonella are estimated to be less than 1% and 0.1%-0.2%, respectively (Kato et al., 2012). Salmonella enterica serotype Choleraesuis is a nontyphoidal Salmonella, highly pathogenic in humans, usually causing septicemic disease with little or no intestinal involvement. Serotype Choleraesuis accounts for a small percentage of published studies of Salmonella infections in the United States. It is not commonly reported in joint fluid and bones in contrast to serotype Enteritidis and Typhi, where a considerable number of cases have been published. Chen et al. in Taiwan found that 21% of bacteremic patients with this infection subsequently develop focal infections such as septic arthritis, pneumonia, peritonitis, and cutaneous abscess (Chen et al., 1999, Chiu et al., 2004). In contrast, our patient presented with localized osteoarticular infection with Salmonella enterica serotype Cholerasuis, but without evidence of bacteremia. PMID:23781356

  18. Serotypes and Antimicrobial Resistance of Human Nontyphoidal Isolates of Salmonella enterica from Crete, Greece

    PubMed Central

    Maraki, Sofia; Papadakis, Ioannis S.

    2014-01-01

    We report on the serotype distribution and the antimicrobial resistance patterns to 20 different antimicrobials of 150 Salmonella enterica strains isolated from stools of diarrhoeal patients on the island of Crete over the period January 2011-December 2012. Among the S. enterica serotypes recovered, Enteritidis was the most prevalent (37.3%), followed by Typhimurium (28.7%) and Newport (8.7%). No resistance was detected to extended-spectrum cephalosporins and carbapenems. Rates of resistance to ampicillin, amoxicillin/clavulanic acid, chloramphenicol, tetracycline, and cotrimoxazole were 9.3%, 4%, 2%, 15.3%, and 8.7%, respectively. Resistance to ≥4 antibiotics was primarily observed for serotypes Typhimurium and Hadar. Enteritidis remains the predominant serotype in Crete. Although low resistance to most antimicrobials was detected, continued surveillance of susceptibility is needed due to the risk of resistance. PMID:24860606

  19. Predicting Salmonella enterica serotypes by repetitive sequence-based PCR

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Repetitive extragenic palindromic sequence-based PCR (rep-PCR) utilizing a semi-automated system, was evaluated as a method to determine Salmonella serotypes. A group of 216 Salmonella isolates belonging to 13 frequently isolated serotypes and one rarer serotype from poultry were used to create a D...

  20. Genomic epidemiology of Salmonella enterica serotype Enteritidis based on population structure of prevalent lineages

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Salmonella enterica serotype Enteritidis (SE) is one of the most commonly reported causes of human salmonellosis. The low genetic diversity of SE measured by fingerprinting methods has made subtyping a challenge. In this study, we used whole genome sequencing to characterize a total of 125 SE and Sa...

  1. Invasive Salmonella enterica serotype typhimurium infections, Democratic Republic of the Congo, 2007-2011.

    PubMed

    Ley, Benedikt; Le Hello, Simon; Lunguya, Octavie; Lejon, Veerle; Muyembe, Jean-Jacques; Weill, François-Xavier; Jacobs, Jan

    2014-04-01

    Infection with Salmonella enterica serotype Typhimurium sequence type (ST) 313 is associated with high rates of drug resistance, bloodstream infections, and death. To determine whether ST313 is dominant in the Democratic Republic of the Congo, we studied 180 isolates collected during 2007-2011; 96% belonged to CRISPOL type CT28, which is associated with ST313. PMID:24655438

  2. Salmonella enterica serotype enteritidis in French Polynesia, South Pacific, 2008-2013.

    PubMed

    Le Hello, Simon; Maillard, Fiona; Mallet, Henri-Pierre; Daudens, Elise; Levy, Marc; Roy, Valérie; Branaa, Philippe; Bertrand, Sophie; Fabre, Laetitia; Weill, François-Xavier

    2015-06-01

    Outbreaks of Salmonella enterica serotype Enteritidis infections associated with eggs occurred in French Polynesia during 2008-2013. Molecular analysis of isolates by using clustered regularly interspaced short palindromic repeat polymorphisms and multilocus variable-number tandem-repeat analysis was performed. This subtyping made defining the epidemic strain, finding the source, and decontaminating affected poultry flocks possible. PMID:25988406

  3. Novel Surveillance of Salmonella enterica Serotype Heidelberg Epidemics in a Closed Community

    Technology Transfer Automated Retrieval System (TEKTRAN)

    During 2003-2005, a systematic and regularly timed human and farm-animal wastewater sampling scheme existed in several prison units in Texas. In early July 2003, an outbreak of gastroenteritis caused by Salmonella enterica serotype Heidelberg occurred in the human population at one site. Wastewate...

  4. PRESENCE OF ANTIBIOTIC RESISTANT GENES IN SALMONELLA ENTERICA SEROTYPE UGANDA ISLOATES FROM 1997-2000

    Technology Transfer Automated Retrieval System (TEKTRAN)

    In the poultry and meat industry, Salmonella enterica serotype Uganda is rarely isolated in the environment. However, recent reports from veterinary diagnostic laboratories indicate an increased frequency of recovery of S. Uganda. Between 1997 and 2000, the animal arm of the National Antimicrobial...

  5. Rapid Molecular Determination of Serotype from Clinical Isolates of Salmonella Enterica

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Background: The conventional serotyping of Salmonella Enterica is time consuming, costly, and requires highly skilled staff. In the present study, we report a multiplex PCR typing method using capillary electrophoresis for fragment analysis that allows for the identification of the 30 most common h...

  6. Complete Closed Genome Sequences of Salmonella enterica subsp. enterica Serotypes Anatum, Montevideo, Typhimurium, and Newport, Isolated from Beef, Cattle, and Humans.

    PubMed

    Harhay, Dayna M; Bono, James L; Smith, Timothy P L; Fields, Patricia I; Dinsmore, Blake A; Santovenia, Monica; Kelley, Christy M; Wang, Rong; Harhay, Gregory P

    2016-01-01

    Salmonella enterica spp. are a diverse group of bacteria with a wide range of virulence potential. To facilitate genome comparisons across this virulence spectrum, we present eight complete closed genome sequences of four S. enterica serotypes (Anatum, Montevideo, Typhimurium, and Newport), isolated from various cattle samples and from humans. PMID:26847891

  7. Complete closed genome sequences of Salmonella enterica subsp. enterica serotypes Anatum, Montevideo, Typhimurium and Newport, isolated from beef, cattle, and humans

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Salmonella enterica are a versatile group of bacteria with a wide range in virulence potential. To facilitate genome comparisons across this virulence spectrum, we present eight complete closed genome sequences of four S. enterica serotypes (Anatum, Montevideo, Typhimurium, and Newport) isolated fro...

  8. Salmonella enterica Serotype Dublin Infection: an Emerging Infectious Disease for the Northeastern United States

    PubMed Central

    McDonough, Patrick L.; Fogelman, David; Shin, Sang J.; Brunner, Michael A.; Lein, Donald H.

    1999-01-01

    Salmonella enterica subspecies enterica serotype Dublin (S. enterica Dublin) emerged for the first time in New York, Pennsylvania, and Ohio in 1988. Since that time this host-adapted serotype has spread throughout the veal- and dairy beef-raising operations in the region; very few dairy farms have experienced clinical S. enterica Dublin infections. This study details the epidemiology of the outbreaks in cattle. During the period 1988 through 1995, nine New York and four Pennsylvania counties have been affected; 13 different locations were involved in New York, and 10 were involved in Pennsylvania. The morbidity and mortality and seasonal distribution of outbreaks, which totaled 35, is described. The antimicrobial susceptibility pattern of isolates revealed that many of the strains were resistant to a number of commonly used drugs. Clinical case details and pathology information are provided, with a caution to clinicians and microbiologists presented with suspect animals, i.e., most cases occurred in older calves, which is atypical for salmonellosis for this region (calves were 8 or more weeks old) and presented as pneumonia and septicemia rather than the primarily diarrheal syndrome that is more typically recognized for the region. The epidemiology of cases is analyzed through cluster analysis of bacterial isolates and their fatty acid methyl ester profiles; at least six clones appeared in the region during the study period. Results of the epidemiology analysis are used to support a hypothesis regarding the source of S. enterica Dublin for the region and its manner of dissemination. PMID:10405378

  9. Analysis of Molecular Epidemiology of Chilean Salmonella enterica Serotype Enteritidis Isolates by Pulsed-Field Gel Electrophoresis and Bacteriophage Typing

    PubMed Central

    Fernandez, Jorge; Fica, Alberto; Ebensperger, German; Calfullan, Hector; Prat, Soledad; Fernandez, Alda; Alexandre, Marcela; Heitmann, Ingrid

    2003-01-01

    Human Salmonella enterica serotype Enteritidis infections emerged in Chile in 1994. S. enterica serotype Enteritidis phage type 1 isolates predominated in the north, and phage type 4 isolates predominated in the central and southern regions. A study was planned to characterize this epidemic using the best discriminatory typing technique. Research involved 441 S. enterica serotype Enteritidis isolates, including clinical preepidemic samples (n = 74; 1975 to 1993) and epidemic (n = 199), food (n = 72), poultry (n = 57), and some Latin American (n = 39) isolates. The best method was selected based on a sample of preepidemic isolates, analyzing the discriminatory power (DP) obtained by phage typing and randomly amplified polymorphic DNA and pulsed-field gel electophoresis (PFGE) analysis. The highest DP was associated with BlnI PFGE-bacteriophage typing analysis (0.993). A total of 38 BlnI patterns (B patterns) were identified before the epidemic period, 19 since 1994, and only 4 in both periods. Two major clusters were identified by phylogenetic analysis, and the predominant B patterns clustered in the same branch. Combined analysis revealed that specific B pattern-phage type combinations (subtypes) disappeared before 1994, that different genotypes associated with S. enterica serotype Enteritidis phage type 4 had been observed since 1988, and that strain diversity increased before the expansion of S. enterica serotype Enteritidis in 1994. Predominant subtype B3-phage type 4 was associated with the central and southern regions, and subtype B38-phage type 1 was associated with the north (P < 0.0001). Food and poultry isolates matched the predominant S. enterica serotype Enteritidis subtypes, but isolates identified in neighboring countries (Peru and Bolivia) did not match S. enterica serotype Enteritidis subtypes identified in the north of Chile. The results of this work demonstrate that genetic diversity, replacement, and expansion of specific S. enterica serotype

  10. Analysis of molecular epidemiology of Chilean Salmonella enterica serotype enteritidis isolates by pulsed-field gel electrophoresis and bacteriophage typing.

    PubMed

    Fernandez, Jorge; Fica, Alberto; Ebensperger, German; Calfullan, Hector; Prat, Soledad; Fernandez, Alda; Alexandre, Marcela; Heitmann, Ingrid

    2003-04-01

    Human Salmonella enterica serotype Enteritidis infections emerged in Chile in 1994. S. enterica serotype Enteritidis phage type 1 isolates predominated in the north, and phage type 4 isolates predominated in the central and southern regions. A study was planned to characterize this epidemic using the best discriminatory typing technique. Research involved 441 S. enterica serotype Enteritidis isolates, including clinical preepidemic samples (n = 74; 1975 to 1993) and epidemic (n = 199), food (n = 72), poultry (n = 57), and some Latin American (n = 39) isolates. The best method was selected based on a sample of preepidemic isolates, analyzing the discriminatory power (DP) obtained by phage typing and randomly amplified polymorphic DNA and pulsed-field gel electophoresis (PFGE) analysis. The highest DP was associated with BlnI PFGE-bacteriophage typing analysis (0.993). A total of 38 BlnI patterns (B patterns) were identified before the epidemic period, 19 since 1994, and only 4 in both periods. Two major clusters were identified by phylogenetic analysis, and the predominant B patterns clustered in the same branch. Combined analysis revealed that specific B pattern-phage type combinations (subtypes) disappeared before 1994, that different genotypes associated with S. enterica serotype Enteritidis phage type 4 had been observed since 1988, and that strain diversity increased before the expansion of S. enterica serotype Enteritidis in 1994. Predominant subtype B3-phage type 4 was associated with the central and southern regions, and subtype B38-phage type 1 was associated with the north (P < 0.0001). Food and poultry isolates matched the predominant S. enterica serotype Enteritidis subtypes, but isolates identified in neighboring countries (Peru and Bolivia) did not match S. enterica serotype Enteritidis subtypes identified in the north of Chile. The results of this work demonstrate that genetic diversity, replacement, and expansion of specific S. enterica serotype

  11. Acute Hepatic Necrosis Caused by Salmonella enterica Serotype I 4,5,12:−:1,2 in a Dog

    PubMed Central

    Meiring, Thelma; Grant, Andrew J.; Watson, Penny J.

    2015-01-01

    Acute hepatic necrosis was diagnosed in a dog. Gram staining and fluorescence in situ hybridization identified Salmonella enterica in the liver, subsequently confirmed as S. enterica serotype I 4,5,12:−:1,2. This is the first report of acute hepatic necrosis with liver failure caused by Salmonella in a dog. PMID:26292301

  12. Intergenic Sequence Ribotyping using a region neighboring dkgB links genovar to Kauffman-White serotype of Salmonella enterica

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Intergenic Sequence Ribotyping using a region neighboring dkgB links genovar to Kauffman-White serotype of Salmonella enterica Previous research identified that the 5S ribosomal (rrn) gene and associated flanking sequences that are closely linked to the dkgB gene of Salmonella enterica were highly ...

  13. Molecular Properties of Salmonella enterica Serotype Paratyphi B Distinguish between Its Systemic and Its Enteric Pathovars

    PubMed Central

    Prager, Rita; Rabsch, Wolfgang; Streckel, Wiebke; Voigt, Wolfgang; Tietze, Erhardt; Tschäpe, Helmut

    2003-01-01

    Salmonella enterica serotype O1,4,5,12:Hb:1,2, designated according to the current Kauffmann-White scheme as S. enterica serotype Paratyphi B, is a very diverse serotype with respect to its clinical and microbiological properties. PCR and blot techniques, which identify the presence, polymorphism, and expression of various effector protein genes, help to distinguish between strains with systemic and enteric outcomes of disease. All serotype Paratyphi B strains from systemic infections have been found to be somewhat genetically related with respect to the pattern of their virulence genes sopB, sopD, sopE1, avrA, and sptP as well as other molecular properties (multilocus enzyme electrophoresis type, pulsed-field gel electrophoresis [PFGE] type, ribotype, and IS200 type). They have been classified as members of the systemic pathovar (SPV). All these SPV strains possess a new sopE1-carrying bacteriophage (designated ΦSopE309) with high SopE1 protein expression but lack the commonly occurring avrA determinant. They exhibit normal SopB protein expression but lack SopD protein production. In contrast, strains from enteric infections classified as belonging to the enteric pathovar possess various combinations of the respective virulence genes, PFGE pattern, and ribotypes. We propose that the PCR technique for testing for the presence of the virulence genes sopE1 and avrA be used as a diagnostic tool for identifying both pathovars of S. enterica serotype Paratyphi B. This will be of great public health importance, since strains of serotype Paratyphi B have recently reemerged worldwide. PMID:12958256

  14. Regional distribution of two dairy-associated Salmonella enterica serotypes

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Salmonella enterica is a zoonotic pathogen frequently associated with dairy farms. The organism can cause disease in cows but is also frequently shed in large numbers by dairy cows that are asymptomatic. Cows on a ~100 head dairy farm in Pennsylvania, USA, (focal dairy) were previously shown to have...

  15. A case of extended spectrum beta-lactamase producing Salmonella enterica serotype paratyphi A from India.

    PubMed

    Roy, Priyamvada; Rawat, Deepti; Malik, Sonia

    2015-01-01

    Enteric fever caused by Salmonella enterica is a systemic infection with high rates of morbidity and mortality. Increasing antibiotic resistance in S. enterica has led to shift in the choice of antibiotics used against this organism from chloramphenicol and ampicillin to trimethoprim-sulfamethoxazole, fluoroquinolones, and extended-spectrum cephalosporins. Resistance to cephalosporins, due to the production of extended-spectrum beta-lactamases (ESBLs), is the cause of serious concern worldwide. So far, these enzymes have been detected in many species of the family Enterobacteriaceae including different serotypes of S. enterica. To the best of our knowledge, however, ESBL production in Salmonella Paratyphi A has not yet been reported from India. We present here a case of ESBL producing Salmonella Paratyphi A from India. This is a worrisome finding with grave clinical implications, since the dissemination of this resistance trait would further limit the therapeutic options available for the treatment of enteric fever. PMID:25673610

  16. Intergenic Sequence Ribotyping using a region neighboring dkgB links genovar to Kauffman-White serotype of Salmonella enterica

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Previous research identified that the 5S ribosomal (rrn) gene and associated flanking sequences that are closely linked to the dkgB gene of Salmonella enterica were highly variable between serotypes, but not between subpopulations within the same serotype (PMID: 17005008). The degree of variability ...

  17. Multidrug-resistant Salmonella enterica serotype Typhi, Gulf of Guinea Region, Africa.

    PubMed

    Baltazar, Murielle; Ngandjio, Antoinette; Holt, Kathryn Elizabeth; Lepillet, Elodie; Pardos de la Gandara, Maria; Collard, Jean-Marc; Bercion, Raymond; Nzouankeu, Ariane; Le Hello, Simon; Dougan, Gordon; Fonkoua, Marie-Christine; Weill, François-Xavier

    2015-04-01

    We identified 3 lineages among multidrug-resistant (MDR) Salmonella enterica serotype Typhi isolates in the Gulf of Guinea region in Africa during the 2000s. However, the MDR H58 haplotype, which predominates in southern Asia and Kenya, was not identified. MDR quinolone-susceptible isolates contained a 190-kb incHI1 pST2 plasmid or a 50-kb incN pST3 plasmid. PMID:25811307

  18. Multidrug-Resistant Salmonella enterica Serotype Typhi, Gulf of Guinea Region, Africa

    PubMed Central

    Baltazar, Murielle; Ngandjio, Antoinette; Holt, Kathryn Elizabeth; Lepillet, Elodie; Pardos de la Gandara, Maria; Collard, Jean-Marc; Bercion, Raymond; Nzouankeu, Ariane; Le Hello, Simon; Dougan, Gordon; Fonkoua, Marie-Christine

    2015-01-01

    We identified 3 lineages among multidrug-resistant (MDR) Salmonella enterica serotype Typhi isolates in the Gulf of Guinea region in Africa during the 2000s. However, the MDR H58 haplotype, which predominates in southern Asia and Kenya, was not identified. MDR quinolone-susceptible isolates contained a 190-kb incHI1 pST2 plasmid or a 50-kb incN pST3 plasmid. PMID:25811307

  19. Predicting Salmonella enterica subsp. enterica Serotypes by Repetitive Extragenic Palindromic Sequence-Based PCR

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The DiversiLabTM System, which employs repetitive extragenic palindromic sequence-based PCR (rep-PCR) to genotype microorganisms, was evaluated as a method to predict the serotype of Salmonella isolates. Two hundred and thirty-three Salmonella isolates belonging to 14 frequently isolated serotypes f...

  20. Profile of Salmonella enterica subsp. enterica (Subspecies I) Serotype 4,5,12:i:− Strains Causing Food-Borne Infections in New York City

    PubMed Central

    Agasan, Alice; Kornblum, John; Williams, George; Pratt, Chi-Chi; Fleckenstein, Phylis; Wong, Marie; Ramon, Alex

    2002-01-01

    Strains of newly emerging Salmonella enterica subsp. enterica (subspecies I) serotype 4,5,12:i:− causing food-borne infections, including a large food poisoning outbreak (n = 86) characterized by persistent diarrhea (14% bloody), abdominal pain, fever, and headache, were examined. The organisms were found in the stool samples from the patients. The biochemical profile of the organisms is consistent with that of S. enterica subsp. I serotypes, except for decreased dulcitol (13%) and increased inositol (96%) utilization. Twenty-eight percent of the strains showed resistance to streptomycin, sulfonamides, or tetracycline only; all three antimicrobial agents; or these agents either alone or in combination with ampicillin, trimethoprim, and trimethoprim-sulfamethoxazole. None of the serotype 4,5,12:i:− strains showed resistance or decreased susceptibility to chloramphenicol or ciprofloxacin. On pulsed-field gel electrophoresis (PFGE), the strains showed 11 or 12 resolvable genomic fragments with 18 banding patterns and three PFGE profile (PFP) clusters (i.e., PFP/A, PFP/B, and PFP/C). Seventy-five percent of the isolates fingerprinted were closely related (zero to three band differences; similarity [Dice] coefficient, 86 to 100%); 63% of these were indistinguishable from each other (PFP/A1). PFP/A1 was common to all strains from the outbreak and 11 hospital sources. Strains from six other hospitals shared clusters PFP/B and PFP/C. PFP/C4, of the environmental isolate, was unrelated to PFP/A and PFP/B. Nine band differences (similarity coefficient, 61%) were noted between PFP/A1 and PFP/E of the multidrug-resistant S. enterica subsp. enterica serotype Typhimurium definitive type 104 strains. Whether these emerging Salmonella strains represent a monophasic, Dul− variant of serotype Typhimurium or S. enterica subsp. enterica serotype Lagos or a distinct serotype of S. enterica subsp. I is not yet known. Some of the phenotypic and genotypic properties of the serotype

  1. Multilocus Sequence Typing as a Replacement for Serotyping in Salmonella enterica

    PubMed Central

    Zhou, Zhemin; Sangal, Vartul; Krauland, Mary G.; Hale, James L.; Harbottle, Heather; Uesbeck, Alexandra; Dougan, Gordon; Harrison, Lee H.; Brisse, Sylvain

    2012-01-01

    Salmonella enterica subspecies enterica is traditionally subdivided into serovars by serological and nutritional characteristics. We used Multilocus Sequence Typing (MLST) to assign 4,257 isolates from 554 serovars to 1092 sequence types (STs). The majority of the isolates and many STs were grouped into 138 genetically closely related clusters called eBurstGroups (eBGs). Many eBGs correspond to a serovar, for example most Typhimurium are in eBG1 and most Enteritidis are in eBG4, but many eBGs contained more than one serovar. Furthermore, most serovars were polyphyletic and are distributed across multiple unrelated eBGs. Thus, serovar designations confounded genetically unrelated isolates and failed to recognize natural evolutionary groupings. An inability of serotyping to correctly group isolates was most apparent for Paratyphi B and its variant Java. Most Paratyphi B were included within a sub-cluster of STs belonging to eBG5, which also encompasses a separate sub-cluster of Java STs. However, diphasic Java variants were also found in two other eBGs and monophasic Java variants were in four other eBGs or STs, one of which is in subspecies salamae and a second of which includes isolates assigned to Enteritidis, Dublin and monophasic Paratyphi B. Similarly, Choleraesuis was found in eBG6 and is closely related to Paratyphi C, which is in eBG20. However, Choleraesuis var. Decatur consists of isolates from seven other, unrelated eBGs or STs. The serological assignment of these Decatur isolates to Choleraesuis likely reflects lateral gene transfer of flagellar genes between unrelated bacteria plus purifying selection. By confounding multiple evolutionary groups, serotyping can be misleading about the disease potential of S. enterica. Unlike serotyping, MLST recognizes evolutionary groupings and we recommend that Salmonella classification by serotyping should be replaced by MLST or its equivalents. PMID:22737074

  2. Multidrug resistance in Salmonella enterica serotype Typhimurium from humans in France (1993 to 2003).

    PubMed

    Weill, François-Xavier; Guesnier, Françoise; Guibert, Véronique; Timinouni, Mohammed; Demartin, Marie; Polomack, Lucette; Grimont, Patrick A D

    2006-03-01

    The aim of this study was to determine the distribution of the antimicrobial resistance phenotypes (R types), the phage types and XbaI-pulsed-field gel electrophoresis (PFGE) types, the genes coding for resistance to beta-lactams and to quinolones, and the class 1 integrons among a representative sample of Salmonella enterica serotype Typhimurium isolates collected from humans in 2002 through the French National Reference Center for Salmonella (NRC-Salm) network. The trends in the evolution of antimicrobial resistance of serotype Typhimurium were reviewed by using NRC-Salm data from 1993, 1997, 2000, and 2003. In 2002, 3,998 isolates of serotype Typhimurium were registered at the NRC-Salm among 11,775 serotyped S. enterica isolates (34%). The most common multiple antibiotic resistance pattern was resistance to amoxicillin, chloramphenicol, streptomycin and spectinomycin, sulfonamides, and tetracycline (ACSSpSuTe R type), with 156 isolates (48.8%). One isolate resistant to extended-spectrum cephalosporins due to the production of TEM-52 extended-spectrum beta-lactamase was detected (0.3%), and one multidrug-resistant isolate was highly resistant to ciprofloxacin (MIC > 32 mg/liter). We found that 57.2% of the isolates tested belonged to the DT104 clone. The main resistance pattern of DT104 isolates was R type ACSSpSuTe (83.2%). However, evolutionary changes have occurred within DT104, involving both loss (variants of Salmonella genomic island 1) and acquisition of genes for drug resistance to trimethoprim or to quinolones. PFGE profile X1 was the most prevalent (74.5%) among DT104 isolates, indicating the need to use a more discriminatory subtyping method for such isolates. Global data from the NRC-Salm suggested that DT104 was the main cause of multidrug resistance in serotype Typhimurium from humans from at least 1997 to 2003, with a roughly stable prevalence during this period. PMID:16517842

  3. Serotypes and antimicrobial resistance of Salmonella enterica ssp in central Thailand, 2001-2006.

    PubMed

    Sirichote, Pantip; Bangtrakulnonth, Aroon; Tianmanee, Kanokwan; Unahalekhaka, Aekkawat; Oulai, Amonrat; Chittaphithakchai, Patcharee; Kheowrod, Wachirapa; Hendriksen, Rene S

    2010-11-01

    This study was carried out to elucidate the epidemiological trends and antimicrobial susceptibilities against Salmonella serovars among Thai patients and asymptomatic carriers during 2001-2006 in central Thailand. A total of 1,401 human and 260 non-human isolates from various sources were included. The isolates were characterized using serotyping and antimicrobial susceptibility testing. The most common serovars in patients submitting stool samples were S. Weltevreden, S. Stanley, S. Anatum, and S. Rissen. Significantly higher odds ratios were observed in blood samples versus stool sample for S. Choleraesuis, S. Enteritidis, S. Typhimurium, and S. Typhi. Children under five years old suffered the most frequently from gastroenteritis. The patients most commonly infected with an invasive serovar were children and people from 26 to 55 years of age. Antimicrobial susceptibility data revealed that S. Schwarzengrund, S. Choleraesuis, S. Anatum, S. Stanley, S. Rissen, and S. Typhimurium were the most resistant serovars observed. The invasive serovar, S. Choleraesuis was resistant to cefotaxime and norfloxacin. Antimicrobial resistance to cefotaxime, was observed in S. Agona, S. Rissen, S. Typhimurium, S. Anatum, and S. Weltevreden. An alarmingly high frequency of resistance to third generation cephalosporins was observed. We recommend Thai authorities take action in order to prevent spread of resistant S. Choleraesuis and other serovars among animals and humans by enforcing a more strict policy on the use of antimicrobials in food animals. PMID:21329317

  4. High-Throughput CRISPR Typing of Mycobacterium tuberculosis Complex and Salmonella enterica Serotype Typhimurium.

    PubMed

    Sola, Christophe; Abadia, Edgar; Le Hello, Simon; Weill, François-Xavier

    2015-01-01

    Spoligotyping was developed almost 18 years ago and still remains a popular first-lane genotyping technique to identify and subtype Mycobacterium tuberculosis complex (MTC) clinical isolates at a phylogeographic level. For other pathogens, such as Salmonella enterica, recent studies suggest that specifically designed spoligotyping techniques could be interesting for public health purposes. Spoligotyping was in its original format a reverse line-blot hybridization method using capture probes designed on "spacers" and attached to a membrane's surface and a PCR product obtained from clustered regularly interspaced short palindromic repeats (CRISPRs). Cowan et al. and Fabre et al. were the first to propose a high-throughput Spoligotyping method based on microbeads for MTC and S. enterica serotype Typhimurium, respectively. The main advantages of the high-throughput Spoligotyping techniques we describe here are their low cost, their robustness, and the existence (at least for MTC) of very large databases that allow comparisons between spoligotypes from anywhere. PMID:25981468

  5. Acquisition of extended-spectrum cephalosporin- and colistin-resistant Salmonella enterica subsp. enterica serotype Newport by pilgrims during Hajj.

    PubMed

    Olaitan, Abiola Olumuyiwa; Dia, Ndèye Méry; Gautret, Philippe; Benkouiten, Samir; Belhouchat, Khadidja; Drali, Tassadit; Parola, Philippe; Brouqui, Philippe; Memish, Ziad; Raoult, Didier; Rolain, Jean-Marc

    2015-06-01

    Gatherings like the Hajj involving many people who travel from different parts of the world represent a risk for the acquisition and dissemination of infectious diseases. In this study, acquisition of multidrug-resistant (MDR) Salmonella spp. in 2013 Hajj pilgrims from Marseille, France, was investigated. In total, 267 rectal swabs were collected from 129 participants before their departure and after their return from the pilgrimage as well as during the pilgrimage from patients with diarrhoea. Samples were screened for the presence of Salmonella using quantitative real-time PCR and culture. Whole-genome sequencing was performed to characterise one of the isolates, and the mechanism leading to colistin resistance was investigated. Six post-Hajj samples and one sample collected during a diarrhoea episode in Hajj were positive for Salmonella by real-time PCR, with five Salmonella enterica belonging to several serotypes recovered by culture, whereas no pre-Hajj sample was positive. Two of the isolates belonged to the epidemic Newport serotype, were resistant to cephalosporins, gentamicin and colistin, and harboured the bla(CTX-M-2) gene and a 12-nucleotide deletion in the pmrB gene leading to colistin resistance. This study shows that pilgrims acquired Salmonella bacteria, including a novel MDR clone, during the Hajj pilgrimage. This calls for more improved public health surveillance during Hajj because Salmonella is one of the most common diarrhoea-causing bacteria worldwide. Therefore, returning pilgrims could disseminate MDR bacteria worldwide upon returning to their home countries. PMID:25769786

  6. Heterogeneity of Persistence of Salmonella enterica Serotype Senftenberg Strains Could Explain the Emergence of this Serotype in Poultry Flocks

    PubMed Central

    Boumart, Zineb; Roche, Sylvie M.; Lalande, Françoise; Virlogeux-Payant, Isabelle; Hennequet-Antier, Christelle; Menanteau, Pierrette; Gabriel, Irène; Weill, François-Xavier; Velge, Philippe; Chemaly, Marianne

    2012-01-01

    Salmonella enterica serotype Senftenberg (S. Senftenberg) has recently become more frequent in poultry flocks. Moreover some strains have been implicated in severe clinical cases. To explain the causes of this emergence in farm animals, 134 S. Senftenberg isolates from hatcheries, poultry farms and human clinical cases were analyzed. Persistent and non-persistent strains were identified in chicks. The non-persistent strains disappeared from ceca a few weeks post inoculation. This lack of persistence could be related to the disappearance of this serotype from poultry farms in the past. In contrast, persistent S. Senftenberg strains induced an intestinal asymptomatic carrier state in chicks similar to S. Enteritidis, but a weaker systemic infection than S. Enteritidis in chicks and mice. An in vitro analysis showed that the low infectivity of S. Senftenberg is in part related to its low capacity to invade enterocytes and thus to translocate the intestinal barrier. The higher capacity of persistent than non-persistent strains to colonize and persist in the ceca of chickens could explain the increased persistence of S. Senftenberg in poultry flocks. This trait might thus present a human health risk as these bacteria could be present in animals before slaughter and during food processing. PMID:22545136

  7. Evolutionary relationships between known serotypes and unique variants of Salmonella enterica as determined by ISR secondary structure

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Intergenic Sequence Ribotyping (ISR) near the dkgB gene of Salmonella enterica subspecies I genome is now used to assign serotype in a manner that is largely concordant with the historical Kauffman-White-LeMinor (KWL) antibody-based scheme. ISR has a number of advantages over the KWL scheme, and it ...

  8. Mobilization properties of small ColE1-like plasmids carrying kanamycin resistance gene isolated from Salmonella enterica serotypes

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Background: Previously we isolated and characterized various groups of small kanamycin resistance (KanR) ColE1-like plasmids from different serotypes of Salmonella enterica isolates. These plasmids all carried the aph(3)-I gene encoding the aminoglycoside phosphotransferase responsible for the kanam...

  9. Comparison of dkgB-linked Intergenic Sequence Ribotyping to DNA Microarray Hydridization for Assigning Serotype to Salmonella Enterica.

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The Kauffman-White scheme has been used for decades to serotype Salmonella enterica, which is a pervasive and persistent cause of food-borne illness. Analysis of whole genomes of the bacterium has revealed that it is unlikely that the Kauffman-White scheme provides the level of discrimination requir...

  10. Serotypes of Salmonella enterica Present in the Internal Organs of Mice Caught On-farm From 1995 – 1998.

    Technology Transfer Automated Retrieval System (TEKTRAN)

    : Salmonella enterica is a persistent and pervasive pathogen that impacts the safety of the food supply, especially in regards to poultry and poultry products. The house mouse Mus musculus is a recognized risk factor for introduction on-farm. More information is needed about Salmonella serotypes tha...

  11. Trends in Serotype Distribution and Antimicrobial Susceptibility in Salmonella enterica Isolates from Humans in Belgium, 2009 to 2013

    PubMed Central

    Ceyssens, Pieter-Jan; Mattheus, Wesley; Vanhoof, Raymond

    2014-01-01

    The Belgian National Reference Centre for Salmonella received 16,544 human isolates of Salmonella enterica between January 2009 and December 2013. Although 377 different serotypes were identified, the landscape is dominated by S. enterica serovars Typhimurium (55%) and Enteritidis (19%) in a ratio which is inverse to European Union averages. With outbreaks of Salmonella serotypes Ohio, Stanley, and Paratyphi B variant Java as prime examples, 20 serotypes displayed significant fluctuations in this 5-year period. Typhoid strains account for 1.2% of Belgian salmonellosis cases. Large-scale antibiotic susceptibility analyses (n = 4,561; panel of 12 antibiotics) showed declining resistance levels in S. Enteritis and Typhimurium isolates for 8 and 3 tested agents, respectively. Despite low overall resistance to ciprofloxacin (4.4%) and cefotaxime (1.6%), we identified clonal lineages of Salmonella serotypes Kentucky and Infantis displaying rising resistance against these clinically important drugs. Quinolone resistance is mainly mediated by serotype-specific mutations in GyrA residues Ser83 and Asp87 (92.2% not wild type), while an additional ParC_Ser80Ile mutation leads to ciprofloxacin resistance in 95.5% S. Kentucky isolates, which exceeds European averages. Plasmid-mediated quinolone resistance (PMQR) alleles qnrA1 (n = 1), qnrS (n = 9), qnrD1 (n = 4), and qnrB (n = 4) were found in only 3.0% of 533 isolates resistant to nalidixic acid. In cefotaxime-resistant isolates, we identified a broad range of Ambler class A and C β-lactamase genes (e.g., blaSHV-12, blaTEM-52, blaCTX-M-14, and blaCTX-M-15) commonly associated with members of the family Enterobacteriaceae. In conclusion, resistance to fluoroquinolones and cefotaxime remains rare in human S. enterica, but clonal resistant serotypes arise, and continued (inter)national surveillance is mandatory to understand the origin and routes of dissemination thereof. PMID:25385108

  12. Rapid Emergence and Clonal Dissemination of CTX-M-15-Producing Salmonella enterica Serotype Virchow, South Korea.

    PubMed

    Kim, Jin Seok; Yun, Young-Sun; Kim, Soo Jin; Jeon, Se-Eun; Lee, Deog-yong; Chung, Gyung Tae; Yoo, Cheon-Kwon; Kim, Junyoung

    2016-01-01

    The prevalence of cefotaxime-resistant Salmonella enterica serotype Virchow has dramatically increased in South Korea since the first isolation in 2011. Of 68 isolates collected over 10 years, 28 cefotaxime-resistant isolates harbored the bla(CTX-M-15) extended-spectrum β-lactamase gene and were closely related genetically, demonstrating the clonal dissemination of CTX-M-15-producing Salmonella Virchow in South Korea. PMID:26674083

  13. CRISPR Is an Optimal Target for the Design of Specific PCR Assays for Salmonella enterica Serotypes Typhi and Paratyphi A

    PubMed Central

    Fabre, Laetitia; Le Hello, Simon; Roux, Chrystelle; Issenhuth-Jeanjean, Sylvie; Weill, François-Xavier

    2014-01-01

    Background Serotype-specific PCR assays targeting Salmonella enterica serotypes Typhi and Paratyphi A, the causal agents of typhoid and paratyphoid fevers, are required to accelerate formal diagnosis and to overcome the lack of typing sera and, in some situations, the need for culture. However, the sensitivity and specificity of such assays must be demonstrated on large collections of strains representative of the targeted serotypes and all other bacterial populations producing similar clinical symptoms. Methodology Using a new family of repeated DNA sequences, CRISPR (clustered regularly interspaced short palindromic repeats), as a serotype-specific target, we developed a conventional multiplex PCR assay for the detection and differentiation of serotypes Typhi and Paratyphi A from cultured isolates. We also developed EvaGreen-based real-time singleplex PCR assays with the same two sets of primers. Principal findings We achieved 100% sensitivity and specificity for each protocol after validation of the assays on 188 serotype Typhi and 74 serotype Paratyphi A strains from diverse genetic groups, geographic origins and time periods and on 70 strains of bacteria frequently encountered in bloodstream infections, including 29 other Salmonella serotypes and 42 strains from 38 other bacterial species. Conclusions The performance and convenience of our serotype-specific PCR assays should facilitate the rapid and accurate identification of these two major serotypes in a large range of clinical and public health laboratories with access to PCR technology. These assays were developed for use with DNA from cultured isolates, but with modifications to the assay, the CRISPR targets could be used in the development of assays for use with clinical and other samples. PMID:24498453

  14. A comparison of cecal colonization of Salmonella enterica serotype Typhimurium in white leghorn chicks and Salmonella-resistant mice

    PubMed Central

    Sivula, Christine P; Bogomolnaya, Lydia M; Andrews-Polymenis, Helene L

    2008-01-01

    Background Salmonellosis is one of the most important bacterial food borne illnesses worldwide. A major source of infection for humans is consumption of chicken or egg products that have been contaminated with Salmonella enterica serotype Typhimurium, however our knowledge regarding colonization and persistence factors in the chicken is small. Results We compared intestinal and systemic colonization of 1-week-old White Leghorn chicks and Salmonella-resistant CBA/J mice during infection with Salmonella enterica serotype Typhimurium ATCC14028, one of the most commonly studied isolates. We also studied the distribution of wild type serotype Typhimurium ATCC14028 and an isogenic invA mutant during competitive infection in the cecum of 1-week-old White Leghorn chicks and 8-week-old CBA/J mice. We found that although the systemic levels of serotype Typhimurium in both infected animal models are low, infected mice have significant splenomegaly beginning at 15 days post infection. In the intestinal tract itself, the cecal contents are the major site for recovery of serotype Typhimurium in the cecum of 1-week-old chicks and Salmonella-resistant mice. Additionally we show that only a small minority of Salmonellae are intracellular in the cecal epithelium of both infected animal models, and while SPI-1 is important for successful infection in the murine model, it is important for association with the cecal epithelium of 1-week-old chicks. Finally, we show that in chicks infected with serotype Typhimurium at 1 week of age, the level of fecal shedding of this organism does not reflect the level of cecal colonization as it does in murine models. Conclusion In our study, we highlight important differences in systemic and intestinal colonization levels between chick and murine serotype Typhimurium infections, and provide evidence that suggests that the role of SPI-1 may not be the same during colonization of both animal models. PMID:18922185

  15. Proteome of Salmonella enterica serotype Tyhimurium Grown in Low Mg2+/pH Medium

    SciTech Connect

    Shi, Liang; Ansong, Charles; Smallwood, Heather S.; Rommereim, Leah M.; McDermott, Jason E.; Brewer, Heather M.; Norbeck, Angela D.; Taylor, Ronald C.; Gustin, Jean K.; Heffron, Fred; Smith, Richard D.; Adkins, Joshua N.

    2009-09-04

    To determine the impact of a low Mg2+/pH defined growth medium (MgM) on the proteome of Salmonella enterica serotype Typhimurium, we cultured S. Typhimurium cells in the medium under two different conditions termed MgM Shock and MgM Dilution and then comparatively analyzed the bacterial cells harvested from these conditions by a global proteomic approach. Proteomic results showed that MgM Shock and MgM Dilution differentially affected the S. Typhimurium proteome. MgM Shock induced a group of proteins whose induction usually occurred at low O2 level, while MgM Dilution induced those related to the type III secretion system (T3SS) of Salmonella Pathogenicity Island 2 (SPI2) and those involved in thiamine or biotin biosynthesis. The metabolic state of the S. Typhimurium cells grown under MgM Shock condition also differed significantly from that under MgM Dilution condition. Western blot analysis not only confirmed the proteomic results, but also showed that the abundances of SPI2-T3SS proteins SsaQ and SseE and biotin biosynthesis proteins BioB and BioD increased after S. Typhimurium infection of RAW 264.7 macrophages. Deletion of the gene encoding BioB reduced the bacterial ability to replicate inside the macrophages, suggesting a biotin-limited environment encountered by S. Typhimurium within RAW 264.7 macrophages.

  16. Genetic and phenotypic evidence of the Salmonella enterica serotype Enteritidis human-animal interface in Chile.

    PubMed

    Retamal, Patricio; Fresno, Marcela; Dougnac, Catherine; Gutierrez, Sindy; Gornall, Vanessa; Vidal, Roberto; Vernal, Rolando; Pujol, Myriam; Barreto, Marlen; González-Acuña, Daniel; Abalos, Pedro

    2015-01-01

    Salmonella enterica serotype Enteritidis is a worldwide zoonotic agent that has been recognized as a very important food-borne bacterial pathogen, mainly associated with consumption of poultry products. The aim of this work was to determine genotypic and phenotypic evidence of S. Enteritidis transmission among seabirds, poultry and humans in Chile. Genotyping was performed using PCR-based virulotyping, pulse-field gel electrophoresis (PFGE) and multi-locus sequence typing (MLST). Pathogenicity-associated phenotypes were determined with survival to free radicals, acidic pH, starvation, antimicrobial resistance, and survival within human dendritic cells. As result of PCR and PFGE assays, some isolates from the three hosts showed identical genotypic patterns, and through MLST it was determined that all of them belong to sequence type 11. Phenotypic assays show diversity of bacterial responses among isolates. When results were analyzed according to bacterial host, statistical differences were identified in starvation and dendritic cells survival assays. In addition, isolates from seabirds showed the highest rates of resistance to gentamycin, tetracycline, and ampicillin. Overall, the very close genetic and phenotypic traits shown by isolates from humans, poultry, and seabirds suggest the inter-species transmission of S. Enteritidis bacteria between hosts, likely through anthropogenic environmental contamination that determines infection of seabirds with bacteria that are potentially pathogenic for other susceptible organism, including humans. PMID:26029196

  17. Genetic and phenotypic evidence of the Salmonella enterica serotype Enteritidis human-animal interface in Chile

    PubMed Central

    Retamal, Patricio; Fresno, Marcela; Dougnac, Catherine; Gutierrez, Sindy; Gornall, Vanessa; Vidal, Roberto; Vernal, Rolando; Pujol, Myriam; Barreto, Marlen; González-Acuña, Daniel; Abalos, Pedro

    2015-01-01

    Salmonella enterica serotype Enteritidis is a worldwide zoonotic agent that has been recognized as a very important food-borne bacterial pathogen, mainly associated with consumption of poultry products. The aim of this work was to determine genotypic and phenotypic evidence of S. Enteritidis transmission among seabirds, poultry and humans in Chile. Genotyping was performed using PCR-based virulotyping, pulse-field gel electrophoresis (PFGE) and multi-locus sequence typing (MLST). Pathogenicity-associated phenotypes were determined with survival to free radicals, acidic pH, starvation, antimicrobial resistance, and survival within human dendritic cells. As result of PCR and PFGE assays, some isolates from the three hosts showed identical genotypic patterns, and through MLST it was determined that all of them belong to sequence type 11. Phenotypic assays show diversity of bacterial responses among isolates. When results were analyzed according to bacterial host, statistical differences were identified in starvation and dendritic cells survival assays. In addition, isolates from seabirds showed the highest rates of resistance to gentamycin, tetracycline, and ampicillin. Overall, the very close genetic and phenotypic traits shown by isolates from humans, poultry, and seabirds suggest the inter-species transmission of S. Enteritidis bacteria between hosts, likely through anthropogenic environmental contamination that determines infection of seabirds with bacteria that are potentially pathogenic for other susceptible organism, including humans. PMID:26029196

  18. RosE represses Std fimbrial expression in Salmonella enterica serotype Typhimurium

    PubMed Central

    Chessa, Daniela; Winter, Maria G; Nuccio, Sean-Paul; Tükel, Çagla; Bäumler, Andreas J

    2008-01-01

    The Salmonella enterica serotype Typhimurium (S. typhimurium) genome contains a large repertoire of putative fimbrial operons that remain poorly characterized because they are not expressed in vitro. In this study, insertions that induced expression of the putative stdABCD fimbrial operon were identified from a random bank of transposon mutants by screening with immuno-magnetic particles for ligand expression (SIMPLE). Transposon insertions upstream of csgC and lrhA or within dam, setB and STM4463 (renamed rosE) resulted in expression of StdA and its assembly into fimbrial filaments on the cell surface. RosE is a novel negative regulator of Std fimbrial expression as indicated by its repression of a std::lacZ reporter construct and by binding of the purified protein to a DNA region upstream of the stdA start codon. Expression of Std fimbriae in the rosE mutant resulted in increased attachment of S. typhimurium to human colonic epithelial cell lines (T-84 and CaCo-2). A rosE mutant exhibited a reduced ability to compete with virulent S. typhimurium for colonization of murine organs, while no defect was observed when both competing strains carried a stdAB deletion. These data suggest that a tight control of Std fimbrial expression mediated by RosE is required during host pathogen interaction. PMID:18331470

  19. Comparison of a PCR serotyping assay, Check&Trace assay for Salmonella, and Luminex Salmonella serotyping assay for the characterization of Salmonella enterica identified from fresh and naturally contaminated cilantro.

    PubMed

    Jean-Gilles Beaubrun, J; Ewing, L; Jarvis, K; Dudley, K; Grim, C; Gopinath, G; Flamer, M-L; Auguste, W; Jayaram, A; Elmore, J; Lamont, M; McGrath, T; Hanes, D E

    2014-09-01

    Salmonella enterica isolated from fresh cilantro samples collected through the USDA/AMS Microbiological Data Program (MDP) were used to compare a PCR serotyping assay against the Check&Trace assay and the Luminex (BioPlex) Salmonella serotyping assay. The study was conducted to evaluate the effectiveness of the three methods for serotyping Salmonella from both enrichment broth cultures and pure Salmonella cultures. In this investigation, Salmonella spp. serotyping was conducted using 24 h enrichment broth cultures and pure Salmonella cultures from cilantro samples, with the PCR serotyping assay. Conversely, the Check&Trace and Luminex for Salmonella assays required pure cultures for Salmonella serotyping. The cilantro samples contained S. enterica serovar Montevideo, Newport, Saintpaul, and Tennessee, identified by the PCR serotyping assay and Check&Trace for Salmonella, but the Luminex assay only identified two of the four serotypes of the cilantro samples. The anticipated impact from this study is that the PCR serotyping assay provides a time- and cost-effective means for screening, identifying and serotyping Salmonella using DNA extracted from 24 h enrichment cilantro samples. PMID:24929735

  20. An outbreak of multidrug-resistant Salmonella enterica subsp. enterica serotype Typhimurium, DT104L linked to dried anchovy in Singapore.

    PubMed

    Ling, M L; Goh, K T; Wang, G C Y; Neo, K S; Chua, T

    2002-02-01

    Multidrug-resistant Salmonella enterica subsp. enterica serotype Typhimurium DT104L was first reported in Singapore from mid-July to mid-October 2000. Salmonella strains isolated from clinical laboratories were submitted to a reference laboratory for serotyping, phage-typing and pulsed-field gel electrophoresis (PFGE) using XbaI restriction endonuclease. An epidemiological investigation was conducted to determine the source of infection and mode of transmission using a structured questionnaire. A total of 33 cases involving mainly infants and toddlers were detected in the 3-month long outbreak. The outbreak strain was of the R-type ACGSTSu, i.e. resistant to ampicillin, chloramphenicol, gentamicin, streptomycin, tetracycline and sulphonamide. PFGE showed all isolates had an indistinguishable pattern, indicating a common source of infection. Consumption of imported dried anchovy was found to be the vehicle of transmission after adjusting for all confounding variables in the case-control study using stepwise logistic regression (OR 25.6; 95% CI 3.9-167.9; P = 0.001). Imported dried seafood should be properly processed, packed, labelled, and thoroughly cooked to prevent transmission of multidrug-resistant S. Typhimurium. PMID:11895083

  1. Comparative analysis of twenty-four complete aenome aequences of Salmonella enterica Serotypes Anatum, Montevideo, Typhimurium and Newport isolated from ground beef or asymptomatic cattle on farm or at harvest

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Salmonella enterica subsp. enterica are a versatile group of bacteria with a wide range of variation in virulence potential. Complete S. enterica genome sequences available to date are primarily of strains isolated from humans or of serotypes that commonly cause human disease. To facilitate genomic ...

  2. Neural Network Model for Survival and Growth of Salmonella enterica Serotype 8,20:-:z6 in Ground Chicken Thigh Meat during Cold Storage: Extrapolation to Other Serotypes.

    PubMed

    Oscar, T P

    2015-10-01

    Mathematical models that predict the behavior of human bacterial pathogens in food are valuable tools for assessing and managing this risk to public health. A study was undertaken to develop a model for predicting the behavior of Salmonella enterica serotype 8,20:-:z6 in chicken meat during cold storage and to determine how well the model would predict the behavior of other serotypes of Salmonella stored under the same conditions. To develop the model, ground chicken thigh meat (0.75 cm(3)) was inoculated with 1.7 log Salmonella 8,20:-:z6 and then stored for 0 to 8 -8 to 16°C. An automated miniaturized most-probable-number (MPN) method was developed and used for the enumeration of Salmonella. Commercial software (Excel and the add-in program NeuralTools) was used to develop a multilayer feedforward neural network model with one hidden layer of two nodes. The performance of the model was evaluated using the acceptable prediction zone (APZ) method. The number of Salmonella in ground chicken thigh meat stayed the same (P > 0.05) during 8 days of storage at -8 to 8°C but increased (P < 0.05) during storage at 9°C (+0.6 log) to 16°C (+5.1 log). The proportion of residual values (observed minus predicted values) in an APZ (pAPZ) from -1 log (fail-safe) to 0.5 log (fail-dangerous) was 0.939 for the data (n = 426 log MPN values) used in the development of the model. The model had a pAPZ of 0.944 or 0.954 when it was extrapolated to test data (n = 108 log MPN per serotype) for other serotypes (S. enterica serotype Typhimurium var 5-, Kentucky, Typhimurium, and Thompson) of Salmonella in ground chicken thigh meat stored for 0 to 8 days at -4, 4, 12, or 16°C under the same experimental conditions. A pAPZ of ≥0.7 indicates that a model provides predictions with acceptable bias and accuracy. Thus, the results indicated that the model provided valid predictions of the survival and growth of Salmonella 8,20:-:z6 in ground chicken thigh meat stored for 0 to 8 days at -8 to

  3. The evaluation of a PCR-based method for identification of Salmonella enterica serotypes from environmental samples and various food matrices.

    PubMed

    Jean-Gilles Beaubrun, Junia; Cheng, Chorng-Ming; Chen, Kai-Shun; Ewing, Laura; Wang, Hua; Agpaoa, Maria C; Huang, Mei-Chiung J; Dickey, Erin; Du, Jamie M; Williams-Hill, Donna M; Hamilton, Brittany; Micallef, Shirley A; Rosenberg Goldstein, Rachel E; George, Ashish; Joseph, Sam W; Sapkota, Amy R; Jacobson, Andrew P; Tall, Ben D; Kothary, Mahendra H; Dudley, Kim; Hanes, Darcy E

    2012-09-01

    The most commonly used method for serotyping Salmonella spp. is based on the Kaufmann-White scheme, and is composed of serological reactions using antibodies to LPS agglutinins. The multiplex PCR used in this investigation was established by Kim et al. to serotype the 30 most common clinical Salmonella serotypes, as determined by CDC. The PCR assay consists of two five-plex reactions and a single two-plex PCR reaction, based on six genetic loci from Salmonella enterica serotype Typhimurium and four loci from S. enterica serotype Typhi. In this investigation, we further evaluated the method for serotyping Salmonella spp. using a reference collection, environmental samples collected from a Mid-Atlantic region tomato farm study, four food matrices spiked with different Salmonella serotypes and a proficiency test. The PCR assay was first evaluated using DNA isolated from pure cultures of isolates obtained from various clinical and environmental samples, and then DNA isolated from broth cultures of food matrices of "Red round" and Roma tomatoes, Romaine lettuce, green onions and Serrano peppers spiked with serotypes Newport, Typhimurium, Javiana and Saintpaul, respectively. The results showed that the PCR assay correctly serotyped Salmonella spp. from the clinical, environmental, spiked food matrices, and proficiency test samples. These findings are significant because the PCR assay was successful in the identification of Salmonella in the spiked samples in a broth culture containing other non-salmonella organism. This method may be a useful resource for the food safety community. PMID:22608224

  4. CTX-M-27 Producing Salmonella enterica Serotypes Typhimurium and Indiana Are Prevalent among Food-Producing Animals in China

    PubMed Central

    Zhang, Wen-Hui; Lin, Xiang-Yan; Xu, Liang; Gu, Xi-Xi; Yang, Ling; Li, Wan; Ren, Si-Qi; Liu, Ya-Hong; Zeng, Zhen-Ling; Jiang, Hong-Xia

    2016-01-01

    Salmonella spp. is one of the most important food-borne pathogens causing digestive tract and invasive infections in both humans and animals. Extended-spectrum β-lactamases (ESBLs) especially the CTX-M-type ESBLs are increasingly being reported worldwide and in China. These studies seldom focused on Salmonella isolates from food-producing animals. The aim of this study was to characterize the antimicrobial resistance profiles, serotypes and ESBLs and in particular, CTX-M producing Salmonella isolates from chickens and pigs in China. Salmonella isolates were identified by API20E system and polymerase chain reaction (PCR) assay; serotypes were determined using slide agglutination with hyperimmune sera; antimicrobial susceptibility was tested using the ager dilution method; the prevalence of ESBLs and PMQR genes were screened by PCR; CTX-M-producing isolates were further characterized by conjugation along with genetic relatedness and plasmid replicon type. In total, 159 Salmonella strains were identified, among which 95 strains were Salmonella enterica serovar Typhimurium, 63 strains were S. enterica serovar Indiana, and 1 strain was S. enterica serovar Enteritidis. All of these isolates presented multi-drug resistant phenotypes. Forty-five isolates carried blaCTX-M genes, the most common subtype was CTX-M-27(34), followed by CTX-M-65(7) and CTX-M-14(4). Most blaCTX-M genes were transmitted by non-typeable or IncN/IncFIB/IncP/IncA/C/IncHI2 plasmids with sizes ranging from 80 to 280 kb. In particular, all the 14 non-typeable plasmids were carrying blaCTX-M-27 gene and had a similar size. PFGE profiles indicated that CTX-M-positive isolates were clonally related among the same serotype, whilst the isolates of different serotypes were genetically divergent. This suggested that both clonal spread of resistant strains and horizontal transmission of the resistance plasmids contributed to the dissemination of blaCTX-M-9G-positive Salmonella isolates. The presence and spread

  5. CTX-M-27 Producing Salmonella enterica Serotypes Typhimurium and Indiana Are Prevalent among Food-Producing Animals in China.

    PubMed

    Zhang, Wen-Hui; Lin, Xiang-Yan; Xu, Liang; Gu, Xi-Xi; Yang, Ling; Li, Wan; Ren, Si-Qi; Liu, Ya-Hong; Zeng, Zhen-Ling; Jiang, Hong-Xia

    2016-01-01

    Salmonella spp. is one of the most important food-borne pathogens causing digestive tract and invasive infections in both humans and animals. Extended-spectrum β-lactamases (ESBLs) especially the CTX-M-type ESBLs are increasingly being reported worldwide and in China. These studies seldom focused on Salmonella isolates from food-producing animals. The aim of this study was to characterize the antimicrobial resistance profiles, serotypes and ESBLs and in particular, CTX-M producing Salmonella isolates from chickens and pigs in China. Salmonella isolates were identified by API20E system and polymerase chain reaction (PCR) assay; serotypes were determined using slide agglutination with hyperimmune sera; antimicrobial susceptibility was tested using the ager dilution method; the prevalence of ESBLs and PMQR genes were screened by PCR; CTX-M-producing isolates were further characterized by conjugation along with genetic relatedness and plasmid replicon type. In total, 159 Salmonella strains were identified, among which 95 strains were Salmonella enterica serovar Typhimurium, 63 strains were S. enterica serovar Indiana, and 1 strain was S. enterica serovar Enteritidis. All of these isolates presented multi-drug resistant phenotypes. Forty-five isolates carried bla CTX-M genes, the most common subtype was CTX-M-27(34), followed by CTX-M-65(7) and CTX-M-14(4). Most bla CTX-M genes were transmitted by non-typeable or IncN/IncFIB/IncP/IncA/C/IncHI2 plasmids with sizes ranging from 80 to 280 kb. In particular, all the 14 non-typeable plasmids were carrying bla CTX-M-27 gene and had a similar size. PFGE profiles indicated that CTX-M-positive isolates were clonally related among the same serotype, whilst the isolates of different serotypes were genetically divergent. This suggested that both clonal spread of resistant strains and horizontal transmission of the resistance plasmids contributed to the dissemination of bla CTX-M-9G-positive Salmonella isolates. The presence and

  6. The global establishment of a highly-fluoroquinolone resistant Salmonella enterica serotype Kentucky ST198 strain

    PubMed Central

    Le Hello, Simon; Bekhit, Amany; Granier, Sophie A.; Barua, Himel; Beutlich, Janine; Zając, Magdalena; Münch, Sebastian; Sintchenko, Vitali; Bouchrif, Brahim; Fashae, Kayode; Pinsard, Jean-Louis; Sontag, Lucile; Fabre, Laetitia; Garnier, Martine; Guibert, Véronique; Howard, Peter; Hendriksen, Rene S.; Christensen, Jens P.; Biswas, Paritosh K.; Cloeckaert, Axel; Rabsch, Wolfgang; Wasyl, Dariusz; Doublet, Benoit; Weill, François-Xavier

    2013-01-01

    While the spread of Salmonella enterica serotype Kentucky resistant to ciprofloxacin across Africa and the Middle-East has been described recently, the presence of this strain in humans, food, various animal species (livestock, pets, and wildlife) and in environment is suspected in other countries of different continents. Here, we report results of an in-depth molecular epidemiological study on a global human and non-human collection of S. Kentucky (n = 70). We performed XbaI-pulsed field gel electrophoresis and multilocus sequence typing, assessed mutations in the quinolone resistance-determining regions, detected β-lactam resistance mechanisms, and screened the presence of the Salmonella genomic island 1 (SGI1). In this study, we highlight the rapid and extensive worldwide dissemination of the ciprofloxacin-resistant S. Kentucky ST198-X1-SGI1 strain since the mid-2000s in an increasingly large number of contaminated sources, including the environment. This strain has accumulated an increasing number of chromosomal and plasmid resistance determinants and has been identified in the Indian subcontinent, Southeast Asia and Europe since 2010. The second substitution at position 87 in GyrA (replacing the amino acid Asp) appeared helpful for epidemiological studies to track the origin of contamination. This global study provides evidence leading to the conclusion that high-level resistance to ciprofloxacin in S. Kentucky is a simple microbiological trait that facilitates the identification of the epidemic clone of interest, ST198-X1-SGI1. Taking this into account is essential in order to detect and monitor it easily and to take rapid measures in livestock to ensure control of this infection. PMID:24385975

  7. Multistate outbreak of Salmonella enterica serotype enteritidis infection associated with pet guinea pigs.

    PubMed

    Bartholomew, Michael L; Heffernan, Richard T; Wright, Jennifer G; Klos, Rachel F; Monson, Timothy; Khan, Sofiya; Trees, Eija; Sabol, Ashley; Willems, Robert A; Flynn, Raymond; Deasy, Marshall P; Jones, Benjamen; Davis, Jeffrey P

    2014-06-01

    Salmonella causes about one million illnesses annually in the United States. Although most infections result from foodborne exposures, animal contact is an important mode of transmission. We investigated a case of Salmonella enterica serotype Enteritidis (SE) sternal osteomyelitis in a previously healthy child who cared for two recently deceased guinea pigs (GPs). A case was defined as SE pulsed-field gel electrophoresis (PFGE) XbaI pattern JEGX01.0021, BlnI pattern JEGA26.0002 (outbreak strain) infection occurring during 2010 in a patient who reported GP exposure. To locate outbreak strain isolates, PulseNet and the US Department of Agriculture National Veterinary Service Laboratories (NVSL) databases were queried. Outbreak strain isolates underwent multilocus variable-number tandem repeat analysis (MLVA). Traceback and environmental investigations were conducted at homes, stores, and breeder or broker facilities. We detected 10 cases among residents of eight states and four NVSL GP outbreak strain isolates. One patient was hospitalized; none died. The median patient age was 9.5 (range, 1-61) years. Among 10 patients, two purchased GPs at independent stores, and three purchased GPs at different national retail chain (chain A) store locations; three were chain A employees and two reported GP exposures of unknown characterization. MLVA revealed four related patterns. Tracebacks identified four distributors and 92 sources supplying GPs to chain A, including one breeder potentially supplying GPs to all case-associated chain A stores. All environmental samples were Salmonella culture-negative. A definitive SE-contaminated environmental source was not identified. Because GPs can harbor Salmonella, consumers and pet industry personnel should be educated regarding risks. PMID:24866204

  8. Identification of Novel Factors Involved in Modulating Motility of Salmonella enterica Serotype Typhimurium

    PubMed Central

    Bogomolnaya, Lydia M.; Aldrich, Lindsay; Ragoza, Yuri; Talamantes, Marissa; Andrews, Katharine D.; McClelland, Michael; Andrews-Polymenis, Helene L.

    2014-01-01

    Salmonella enterica serotype Typhimurium can move through liquid using swimming motility, and across a surface by swarming motility. We generated a library of targeted deletion mutants in Salmonella Typhimurium strain ATCC14028, primarily in genes specific to Salmonella, that we have previously described. In the work presented here, we screened each individual mutant from this library for the ability to move away from the site of inoculation on swimming and swarming motility agar. Mutants in genes previously described as important for motility, such as flgF, motA, cheY are do not move away from the site of inoculation on plates in our screens, validating our approach. Mutants in 130 genes, not previously known to be involved in motility, had altered movement of at least one type, 9 mutants were severely impaired for both types of motility, while 33 mutants appeared defective on swimming motility plates but not swarming motility plates, and 49 mutants had reduced ability to move on swarming agar but not swimming agar. Finally, 39 mutants were determined to be hypermotile in at least one of the types of motility tested. Both mutants that appeared non-motile and hypermotile on plates were assayed for expression levels of FliC and FljB on the bacterial surface and many of them had altered levels of these proteins. The phenotypes we report are the first phenotypes ever assigned to 74 of these open reading frames, as they are annotated as ‘hypothetical genes’ in the Typhimurium genome. PMID:25369209

  9. A new therapeutic challenge for old pathogens: community-acquired invasive infections caused by ceftriaxone- and ciprofloxacin-resistant Salmonella enterica serotype choleraesuis.

    PubMed

    Ko, Wen-Chien; Yan, Jing-Joung; Yu, Wen-Liang; Lee, Hsin-Chun; Lee, Nan-Yao; Wang, Li-Rong; Chuang, Yin-Ching

    2005-01-15

    Recently, antimicrobial resistance among nontyphoid Salmonella serotypes has been increasingly recognized. In southern Taiwan, we encountered 3 cases of invasive infections caused by Salmonella enterica serotype Choleraesuis with resistance to ciprofloxacin and ceftriaxone. Resistance to ciprofloxacin was related to nucleotide mutations in gyrA and parC, and resistance to ceftriaxone was related to the presence of CMY-2 beta -lactamase. PMID:15655754

  10. An outbreak of Salmonella enterica serotype Choleraesuis in goitered gazelle (Gazella subgutrosa subgutrosa) and a Malayan tapir (Tapirus indicus).

    PubMed

    Wolf, Tiffany M; Wünschmann, Arno; Morningstar-Shaw, Brenda; Pantlin, Gayle C; Rasmussen, James M; Thompson, Rachel L

    2011-12-01

    An outbreak of Salmonella enterica serotype Choleraesuis enteritis occurred in two juvenile goitered gazelles and an adult Malayan tapir over a period of 5 wk at the Minnesota Zoo. Diagnosis was made postmortem on one gazelle and one tapir, and a second gazelle was diagnosed via fecal culture. The death of the tapir was attributed to S. enterica serovar Choleraesuis septicemia, while salmonellosis was considered to be a contributing factor besides ostertagiasis for the death of one goitered gazelle and for the diarrhea of another goitered gazelle. A third gazelle became ill in the same time period, but Salmonella infection was not confirmed by culture. All exhibited the clinical signs of profuse, watery diarrhea. The gazelles developed a protein-losing enteropathy, and the tapir showed signs of sepsis and endotoxemia. Serotyping and pulsed-field gel electrophoresis revealed the Salmonella isolates to be indistinguishable from each other. One year prior to this outbreak, Salmonella sp. was cultured from a Visayan warty pig (Sus cebifrons) housed in the same building as the tapir. After further investigation into the outbreak, spread of this pathogen was speculated to be associated with human movement across animal areas. PMID:22204065

  11. PCR Detection of Salmonella enterica Serotype Montevideo in and on Raw Tomatoes Using Primers Derived from hilA

    PubMed Central

    Guo, Xuan; Chen, Jinru; Beuchat, Larry R.; Brackett, Robert E.

    2000-01-01

    Salmonellae have been some of the most frequently reported etiological agents in fresh-produce-associated outbreaks of human infections in recent years. PCR assays using four innovative pairs of primers derived from hilA and sirA, positive regulators of Salmonella invasive genes, were developed to identify Salmonella enterica serotype Montevideo on and in tomatoes. Based on examination of 83 Salmonella strains and 22 non-Salmonella strains, we concluded that a pair of hilA primers detects Salmonella specifically. The detection limits of the PCR assay were 101 and 100 CFU/ml after enrichment at 37°C for 6 and 9 h, respectively. When the assay was validated by detecting S. enterica serotype Montevideo in and on artificially inoculated tomatoes, 102 and 101 CFU/g were detected, respectively, after enrichment for 6 h at 37°C. Our results suggest that the hilA-based PCR assay is sensitive and specific, and can be used for rapid detection of Salmonellae in or on fresh produce. PMID:11097898

  12. Salmonella enterica Serovar Szentes, a Rare Serotype Causing a 9-Month Outbreak in 2013 and 2014 in Switzerland.

    PubMed

    Nüesch-Inderbinen, Magdalena; Cernela, Nicole; Althaus, Denise; Hächler, Herbert; Stephan, Roger

    2015-11-01

    During the summer of 2013, an increase of Salmonella enterica ssp. enterica serovar Szentes isolates from human clinical cases was registered by the Swiss National Centre for Enteropathogenic Bacteria and Listeria. In the course of the ensuing 9 months, 18 isolates originating from 13 patients and from one food sample were collected. Of the 13 human cases, 10 (77%) were female. The patients' ages ranged from 27 to 83 years (median age 49 years). Pulsed-field gel electrophoresis (PFGE) performed with XbaI, and multilocus sequence typing (MLST) were used to type the strains. PFGE as well as MLST showed the strains as indistinguishable. The PFGE pattern and MLST sequence type (ST427) were identical to those of Salmonella enterica serovar Szentes isolated in previous years (2002-2013) from sporadic cases in Switzerland and Germany. The increased isolation frequency continued for 6 months after the detection of Salmonella Szentes in sprouts. No common food exposure could be established. Due to lack of information on the potential food source, further investigations were not possible. The outbreak of this unusual serotype was detected because of its temporal clustering. PMID:26287690

  13. Characterization of Anti-Salmonella enterica Serotype Typhi Antibody Responses in Bacteremic Bangladeshi Patients Using Immuno-affinity Proteomic-based Technology (IPT)

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Salmonella enterica serotype Typhi (S. Typhi) is the cause of typhoid fever and a human-restricted pathogen. Currently available typhoid vaccines provide only 50-75% protection for 2-5 years, and available diagnostic assays to identify individuals with typhoid fever lack both sensitivity and specifi...

  14. Nalidixic Acid-Resistant Salmonella enterica Serotype Typhi Presenting as a Primary Psoas Abscess: Case Report and Review of the Literature

    PubMed Central

    Shakespeare, William A.; Davie, Daniel; Tonnerre, Claude; Rubin, Michael A.; Strong, Michael; Petti, Cathy A.

    2005-01-01

    We report an unusual case of Salmonella enterica serotype Typhi presenting as a primary psoas abscess. The isolate tested susceptible to ciprofloxacin but resistant to nalidixic acid in vitro, a pattern associated with fluoroquinolone therapeutic failures. We review the literature for serovar Typhi psoas abscess in the absence of bacteremia and discuss the importance of identifying isolates with reduced susceptibility to fluoroquinolones. PMID:15695728

  15. HIGH PREVALENCE OF INCOMPATIBILITY GROUP INCA/C AMONG MULTI-DRUG RESISTANT SALMONELLA ENTERICA SEROTYPE NEWPORT ISOLATED FROM DAIRY CATTLE

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Multi-drug resistant (MDR) Salmonella enterica serotype Newport exhibiting increased morbidity and mortality among man and animals has emerged in North America and Canada. Risk factors for infection by MDR S. Newport have included exposure to dairy farms, consumption of undercooked ground beef or u...

  16. Characterization of Anti-Salmonella enterica Serotype Typhi Antibody Responses in Bacteremic Bangladeshi Patients by an Immuno-affinity Proteomic-based Technology (IPT)

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Salmonella enterica serotype Typhi (S. Typhi) is the cause of typhoid fever and a human-restricted pathogen. Currently available typhoid vaccines provide only 50-75% protection for 2-5 years, and available diagnostic assays to identify individuals with typhoid fever lack both sensitivity and specif...

  17. Development of a High-Throughput Multiplex PCR and Capillary Electrophoresis Technique for Serotype Determination of Salmonella Enterica Food Animal Isolates

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Background: Previously, a multiplex PCR technique was developed to identify the top 30 human clinical serotypes of Salmonella enterica. To improve the speed, ease of use, utility and discriminatory ability of the technique, additional primers were added and the PCR product discrimination and analysi...

  18. Sensitivity of mycobacterium avium subsp paratuberculosis, escherichia coli and salmonella enterica serotype typhimurium to low pH, high organic acids and ensiling

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The ability of Mycobacterium avium subsp paratuberculosis (M. paratuberculosis), Salmonella enterica serotype Typhimurium (S. Typhimurium) and a commensal Escherichia coli (E. coli) isolate to persist under low pH and high organic acid conditions was determined. Die-off rates were calculated followi...

  19. Antimicrobial susceptibility profiles of Salmonella enterica serotypes recovered from pens of commercial feedlot cattle using different types of composite samples.

    PubMed

    Alam, Mohammad Jahangir; Renter, David; Taylor, Ethel; Mina, Diana; Moxley, Rodney; Smith, David

    2009-04-01

    Salmonella enterica in cattle production systems may be associated with important human and animal disease issues. However, tremendous diversity exists among Salmonella recovered, and more information is needed about strains of greatest potential health concern, particularly those that are multidrug resistant (MDR). By characterizing Salmonella isolates from commercial feedlot pens, this study aimed to evaluate the strain diversity and prevalence of MDR Salmonella from different types of composite pen samples. Antimicrobial susceptibility profiles, serotype, and presence or absence of the integron-encoded intI1 gene were determined for 530 Salmonella isolates recovered using composite rope (n = 335), feces (n = 59), and water (n = 136) samples from 21 pens in 3 feedlots. The study investigated only pens with available isolates from multiple sample types. Most isolates (83.0%) of the 19 Salmonella serotypes identified were susceptible or intermediately susceptible to all the antimicrobials evaluated. Resistance to sulfisoxazole (14.9%), streptomycin (3.8%), and tetracycline (3.6%) were the most common. None of the isolates tested positive for a class 1 integron, and only 2.5% were resistant to multiple antimicrobials. All the MDR isolates, namely, serotypes Uganda (n = 9), Typhimurium (n = 2), and Give (n = 2), were resistant to at least five antimicrobials. Most MDR isolates (n = 11) were from two pens during 1 week within one feedlot. Overall, many Salmonella isolates collected within a pen were similar in terms of serotype and antimicrobial susceptibility regardless of sample type. However, MDR Salmonella and rare serotypes were not recovered frequently enough to suggest a general strategy for appropriate composite sampling of feedlot cattle populations for Salmonella detection and monitoring. PMID:19219500

  20. Diversity and molecular variation among plasmids in Salmonella enterica serotype Dublin based on restriction enzyme fragmentation pattern analysis.

    PubMed Central

    Browning, L. M.; Wray, C.; Platt, D. J.

    1995-01-01

    Molecular variation within and between plasmids of Salmonella enterica serotype Dublin was analysed. Such variation has been demonstrated in the serotype-specific plasmids (SSP's) of Typhimurium and Enteritidis. The two aims of this study were to determine the plasmid diversity in a host-adapted serotype and also the incidence of molecular variation in the SSP among strains of Dublin using restriction endonuclease fragmentation pattern (REFP) analysis with Pst1, Sma1 and EcoRV. Sixty-five strains were examined from seven countries. Plasmid profile and REFP analysis showed that none of the strains was plasmid-free. Seventy-seven percent of the strains possessed the 72 kb SSP either alone or in combination with another plasmid; 23% harboured plasmids which were molecular variants of the SSP. Four of the variants were more closely related to each other than to the reference SSP and were harboured by Dublin isolated from both the USA and Europe. A further three were shown to be cointegrate plasmids and were similarly distributed. Thirty-two percent of strains possessed the SSP alone. None of the UK strains was resistant to any of the antimicrobial agents tested whereas 74% of the remaining strains were resistant to between one and five antimicrobial agents. This study corroborates previous findings concerning the high degree of stability of the SSP and confirmed the clonal nature of Dublin. Co-resident plasmids provided evidence of sub-clones within localized geographical areas. Images Fig. 2 PMID:7705487

  1. Genetic diversity and antimicrobial resistance profiles of Salmonella enterica serotype derby isolated from pigs, pork, and humans in France.

    PubMed

    Kerouanton, Annaëlle; Rose, Valérie; Weill, François-Xavier; Granier, Sophie A; Denis, Martine

    2013-11-01

    In France, Salmonella enterica serotypes Typhimurium and Derby are the most often isolated serotypes in pigs. Moreover, serotype Derby usually ranks between third and fourth in prevalence among human isolates in France. The aim of this study was to evaluate the genetic relationships between human and pig Salmonella Derby isolates based on their pulsed-field gel electrophoresis (PFGE) patterns after XbaI, BlnI, and SpeI restriction and on their antimicrobial resistance profiles. The 196 studied isolates were isolated in 2006 and 2007: 73 from fattening pigs, 27 from pork, and 96 from humans. Forty-four PFGE XbaI patterns were identified. A major pattern (SDX01) was identified for 96 isolates (49%). This pattern was common to pig, pork, and human isolates. Among the 146 isolates tested for their antimicrobial resistance, 84.2% (n=123) showed resistance to at least one antibiotic and 69.2% (n=101) were simultaneously resistant to at least streptomycin, sulfonamides, and tetracycline. Most of the isolates that are resistant to these three antibiotics also displayed the major SDX01 pattern. The use of two other restriction enzymes on a part of the panel (155 isolates) brought a significant increase in the discriminatory index, in particular for SDX01 strains. As Salmonella Derby is essentially isolated from pigs, and major resistance and PFGE patterns of isolates from pigs and pork were very similar to human isolates, human salmonellosis due to Salmonella Derby may be related to pigs. PMID:23944749

  2. Identification and characterization of multidrug-resistant Salmonella enterica serotype Albert isolates in the United States

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Salmonella enterica is one of the most common causes of bacterial foodborne illness in the United States. Although most Salmonella infections are self-limiting, antimicrobial treatment is critical for invasive salmonellosis. Primary antimicrobial treatment options include fluoroquinolones or extende...

  3. Host Restriction of Salmonella enterica Serotype Typhimurium Pigeon Isolates Does Not Correlate with Loss of Discrete Genes

    PubMed Central

    Andrews-Polymenis, Helene L.; Rabsch, Wolfgang; Porwollik, Steffen; McClelland, Michael; Rosetti, Carlos; Adams, L. Garry; Bäumler, Andreas J.

    2004-01-01

    The definitive phage types (DT) 2 and 99 of Salmonella enterica serotype Typhimurium are epidemiologically correlated with a host range restricted to pigeons, in contrast to phage types with broader host ranges such as epidemic cattle isolates (DT104 and DT204). To determine whether phage types with broad host range possess genetic islands absent from host-restricted phage types, we compared the genomes of four pigeon isolates to serotype Typhimurium strain LT2 using a DNA microarray. Three of the four isolates tested caused fluid accumulation in bovine ligated ileal loops, but they had reduced colonization of liver and spleen in susceptible BALB/c mice and were defective for intestinal persistence in Salmonella-resistant CBA mice. The genomes of the DT99 and DT2 isolates were extremely similar to the LT2 genome, with few notable differences on the level of complete individual genes. Two large groups of genes representing the Fels-1 and Fels-2 prophages were missing from the DT2 and DT99 phage types we analyzed. One of the DT99 isolates examined was lacking a third cluster of five chromosomal genes (STM1555 to -1559). Results of the microarray analysis were extended using Southern analysis to a collection of 75 serotype Typhimurium clinical isolates of 24 different phage types. This analysis revealed no correlation between the presence of Fels-1, Fels-2, or STM1555 to -1559 and the association of phage types with different host reservoirs. We conclude that serotype Typhimurium phage types with broad host range do not possess genetic islands influencing host restriction, which are absent from the host-restricted pigeon isolates. PMID:15090502

  4. Prevalence, Enumeration, Serotypes, and Antimicrobial Resistance Phenotypes of Salmonella enterica Isolates from Carcasses at Two Large United States Pork Processing Plants

    PubMed Central

    Brichta-Harhay, Dayna M.; Kalchayanand, Norasak; Bosilevac, Joseph M.; Shackelford, Steven D.; Wheeler, Tommy L.; Koohmaraie, Mohammad

    2012-01-01

    The objective of this study was to characterize Salmonella enterica contamination on carcasses in two large U.S. commercial pork processing plants. The carcasses were sampled at three points, before scalding (prescald), after dehairing/polishing but before evisceration (preevisceration), and after chilling (chilled final). The overall prevalences of Salmonella on carcasses at these three sampling points, prescald, preevisceration, and after chilling, were 91.2%, 19.1%, and 3.7%, respectively. At one of the two plants, the prevalence of Salmonella was significantly higher (P < 0.01) for each of the carcass sampling points. The prevalences of carcasses with enumerable Salmonella at prescald, preevisceration, and after chilling were 37.7%, 4.8%, and 0.6%, respectively. A total of 294 prescald carcasses had Salmonella loads of >1.9 log CFU/100 cm2, but these carcasses were not equally distributed between the two plants, as 234 occurred at the plant with higher Salmonella prevalences. Forty-one serotypes were identified on prescald carcasses with Salmonella enterica serotypes Derby, Typhimurium, and Anatum predominating. S. enterica serotypes Typhimurium and London were the most common of the 24 serotypes isolated from preevisceration carcasses. The Salmonella serotypes Johannesburg and Typhimurium were the most frequently isolated serotypes of the 9 serotypes identified from chilled final carcasses. Antimicrobial susceptibility was determined for selected isolates from each carcass sampling point. Multiple drug resistance (MDR), defined as resistance to three or more classes of antimicrobial agents, was identified for 71.2%, 47.8%, and 77.5% of the tested isolates from prescald, preevisceration, and chilled final carcasses, respectively. The results of this study indicate that the interventions used by pork processing plants greatly reduce the prevalence of Salmonella on carcasses, but MDR Salmonella was isolated from 3.2% of the final carcasses sampled. PMID:22327585

  5. A community outbreak of Salmonella enterica serotype Typhimurium associated with an asymptomatic food handler in two local restaurants.

    PubMed

    Holman, Emily J; Allen, Keith S; Holguin, John R; Torno, Mauro; Lachica, Miriam

    2014-09-01

    Between January and April 2012, the city of Long Beach Department of Health and Human Services investigated an outbreak involving 19 case patients who had tested positive for Salmonella enterica serotype Typhimurium with indistinguishable pulsed-field gel electrophoresis patterns. All cases were residents of or traveled to the city of Long Beach, California, during their incubation period, and the majority of patients reported eating at one of two restaurants in Long Beach. This article describes the outbreak investigation that traced the source to an asymptomatic food handler working at both restaurants and highlights the importance of maintaining a high index of suspicion for food handlers when faced with local outbreaks of diarrheal illness. PMID:25226780

  6. Genome Sequence of Salmonella enterica Serotype Tennessee Strain CDC07-0191, Implicated in the 2006-2007 Multistate Food-Borne Outbreak Linked to Peanut Butter in the United States.

    PubMed

    Deng, Xiangyu; Salazar, Joelle K; Frezet, Stephanie; Maccannell, Duncan; Ribot, Efrain M; Fields, Patricia I; Fricke, W Florian; Zhang, Wei

    2013-01-01

    Salmonella enterica serotype Tennessee strain CDC07-0191 was isolated from the 2006-2007 multistate food-borne outbreak linked to peanut butter in the United States. Here we report a high-quality draft assembly of the genome sequence of this strain, derived from a patient. This is the first reported high-quality draft genome sequence for S. enterica serotype Tennessee, which will enable in-depth studies of its transmission and virulence. PMID:23704182

  7. Population Heterogeneity of Salmonella enterica Serotype Typhimurium Resulting from Phase Variation of the lpf Operon In Vitro and In Vivo

    PubMed Central

    Kingsley, Robert A.; Weening, Eric H.; Keestra, A. Marijke; Bäumler, Andreas J.

    2002-01-01

    The lpf fimbrial operon oscillates between phase ON and phase OFF expression states, thereby generating heterogeneity within S. enterica serotype Typhimurium populations with regard to expression of long polar fimbrial antigens. To determine whether the proportion of lpf phase variants changes with growth conditions, the lpf phase ON content of cultures was determined after in vitro and in vivo passage. After passage in Luria-Bertani (LB) broth for 120 generations, 96% of cells in a serotype Typhimurium culture carried the lpf operon in the phase ON expression state, regardless of the phase ON/OFF ratio in the inoculum. In contrast, a culture passaged on LB agar plates for 500 generations contained approximately 2% lpf phase ON cells. Differences in the lpf phase ON content of cultures passaged in broth and on plates were not caused by an outgrowth of lpf phase ON or lpf phase OFF cells, since deletion of lpf biosynthesis genes did not alter the phase ON/OFF ratio attained after passage. Instead, growth in LB broth resulted in a eightfold increase in the phase OFF-to-ON transition frequency and a decrease of the lpf phase ON-to-OFF transition frequency by a factor of 150 compared to growth on LB agar plates. After infection of naïve CBA/J mice with an lpf phase ON culture of serotype Typhimurium, the proportion of lpf phase ON cells continuously decreased over time, regardless of whether the strain carried intact fimbrial biosynthesis genes. These data suggest that elaboration of fimbriae does not have a major influence on the population heterogeneity produced by phase variation of the lpf operon in naïve mice. PMID:11948147

  8. Population heterogeneity of Salmonella enterica serotype Typhimurium resulting from phase variation of the lpf operon in vitro and in vivo.

    PubMed

    Kingsley, Robert A; Weening, Eric H; Keestra, A Marijke; Bäumler, Andreas J

    2002-05-01

    The lpf fimbrial operon oscillates between phase ON and phase OFF expression states, thereby generating heterogeneity within S. enterica serotype Typhimurium populations with regard to expression of long polar fimbrial antigens. To determine whether the proportion of lpf phase variants changes with growth conditions, the lpf phase ON content of cultures was determined after in vitro and in vivo passage. After passage in Luria-Bertani (LB) broth for 120 generations, 96% of cells in a serotype Typhimurium culture carried the lpf operon in the phase ON expression state, regardless of the phase ON/OFF ratio in the inoculum. In contrast, a culture passaged on LB agar plates for 500 generations contained approximately 2% lpf phase ON cells. Differences in the lpf phase ON content of cultures passaged in broth and on plates were not caused by an outgrowth of lpf phase ON or lpf phase OFF cells, since deletion of lpf biosynthesis genes did not alter the phase ON/OFF ratio attained after passage. Instead, growth in LB broth resulted in a eightfold increase in the phase OFF-to-ON transition frequency and a decrease of the lpf phase ON-to-OFF transition frequency by a factor of 150 compared to growth on LB agar plates. After infection of naïve CBA/J mice with an lpf phase ON culture of serotype Typhimurium, the proportion of lpf phase ON cells continuously decreased over time, regardless of whether the strain carried intact fimbrial biosynthesis genes. These data suggest that elaboration of fimbriae does not have a major influence on the population heterogeneity produced by phase variation of the lpf operon in naïve mice. PMID:11948147

  9. What’s in a Name? Species-Wide Whole-Genome Sequencing Resolves Invasive and Noninvasive Lineages of Salmonella enterica Serotype Paratyphi B

    PubMed Central

    Owen, Sian V.; Langridge, Gemma; Connell, Steve; Nair, Satheesh; Reuter, Sandra; Dallman, Timothy J.; Corander, Jukka; Tabing, Kristine C.; Le Hello, Simon; Fookes, Maria; Doublet, Benoît; Zhou, Zhemin; Feltwell, Theresa; Ellington, Matthew J.; Herrera, Silvia; Gilmour, Matthew; Cloeckaert, Axel; Achtman, Mark; Wain, John; De Pinna, Elizabeth; Weill, François-Xavier; Peters, Tansy; Thomson, Nick

    2016-01-01

    ABSTRACT For 100 years, it has been obvious that Salmonella enterica strains sharing the serotype with the formula 1,4,[5],12:b:1,2—now known as Paratyphi B—can cause diseases ranging from serious systemic infections to self-limiting gastroenteritis. Despite considerable predicted diversity between strains carrying the common Paratyphi B serotype, there remain few methods that subdivide the group into groups that are congruent with their disease phenotypes. Paratyphi B therefore represents one of the canonical examples in Salmonella where serotyping combined with classical microbiological tests fails to provide clinically informative information. Here, we use genomics to provide the first high-resolution view of this serotype, placing it into a wider genomic context of the Salmonella enterica species. These analyses reveal why it has been impossible to subdivide this serotype based upon phenotypic and limited molecular approaches. By examining the genomic data in detail, we are able to identify common features that correlate with strains of clinical importance. The results presented here provide new diagnostic targets, as well as posing important new questions about the basis for the invasive disease phenotype observed in a subset of strains. PMID:27555304

  10. Interpretations of Antibody Responses to Salmonella enterica Serotype Enteritidis gm Flagellin in Poultry Flocks Are Enhanced by a Kinetics-Based Enzyme-Linked Immunosorbent Assay

    PubMed Central

    McDonough, Patrick L.; Jacobson, Richard H.; Timoney, John F.; Mutalib, Ahmed; Kradel, David C.; Chang, Yung-fu; Shin, Sang J.; Lein, Donald H.; Trock, Susan; Wheeler, Kaye

    1998-01-01

    Many regulatory and diagnostic programs for the detection of Salmonella enterica serotype Enteritidis infection in commercial poultry flocks have relied on rapid Pullorum agglutination tests to screen birds because of the shared antigens of S. enterica Enteritidis and S. enterica Pullorum and Gallinarum; however, the use of the enzyme-linked immunosorbent assay (ELISA) format affords better analytical sensitivity than crude agglutination tests. In this study, we adapted our earlier conventional indirect ELISA, using gm flagellin as the antigen, to a kinetics-based, computer-controlled ELISA (KELA). The KELA was used to screen for flagellin antibody from three commercial flocks: (i) a large flock involved in a U.S. Department of Agriculture trace back from a human S. enterica Enteritidis foodborne outbreak (n = 3,209), (ii) a flock infected with the endemic S. enterica Enteritidis serotype but which also had multiple other salmonella serotypes (n = 65), and (iii) an S. enterica Pullorum-infected flock (n = 12). The first flock (S. enterica Enteritidis prevalence of 2.45% based on culture) provided a field test of the KELA and allowed the calculation of diagnostic sensitivity (D-Sn) and diagnostic specificity (D-Sp). With a cutoff of 10 (used for screening flocks [i.e., high sensitivity]), the KELA has a D-Sn of 95.2% and a D-Sp of 18.5%; with a cutoff of 140 (used in confirmatory flock testing [i.e., high specificity]), the KELA has a D-Sn of 28.0% and a D-Sp of 99.1%. We found that with a cutoff of 60 (D-Sn = 63.1%; D-Sp = 91.6%), we could eliminate reactions in the KELA caused by other non-S. enterica Enteritidis salmonellae. The KELA was also compared to two commercial rapid Pullorum tests, the Solvay (D-Sn = 94.9%; D-Sp = 55.5%) and the Vineland (D-Sn = 62.0%; D-Sp = 75.3%). PMID:9665965

  11. Salmonella enterica Serotype Gambia with CTX-M-3 and armA Resistance Markers: Nosocomial Infections with a Fatal Outcome▿

    PubMed Central

    Moissenet, D.; Weill, F.-X.; Arlet, G.; Harrois, D.; Girardet, J. P.; Vu-Thien, H.

    2011-01-01

    We report two cases of bacteremia caused by the Salmonella enterica serotype Gambia in our children's hospital, with one fatal outcome. The isolates showed indistinguishable genotypes and infrequent resistance markers: CTX-M-3 extended-spectrum β-lactamase and armA methyltransferase. This is the first report of S. Gambia exhibiting CTX-M-3 and armA markers involved in serious infections. PMID:21270227

  12. SHV-12-Like Extended-Spectrum-β-Lactamase-Producing Strains of Salmonella enterica Serotypes Babelsberg and Enteritidis Isolated in France among Infants Adopted from Mali

    PubMed Central

    Weill, François-Xavier; Demartin, Marie; Tandé, Didier; Espié, Emmanuelle; Rakotoarivony, Ignace; Grimont, Patrick A. D.

    2004-01-01

    From December 2002 to June 2003, 14 cultures of Salmonella enterica serotype Babelsberg and 6 cultures of serotype Enteritidis, isolated in France from internationally adopted children, were identified at the French National Reference Center for Salmonella. All serotype Babelsberg isolates were related, as determined by pulsed-field gel electrophoresis, and all serotype Enteritidis strains displayed the same phage type. All serotype Enteritidis and seven serotype Babelsberg isolates produced an SHV-12-like extended-spectrum β-lactamase as determined by sequencing of PCR products and by isoelectrofocusing. Some serotype Enteritidis isolates exhibited additional antimicrobial resistance (aminoglycosides, tetracycline, chloramphenicol, sulfonamides, and trimethoprim). Our investigation indicated that these Salmonella isolates were certainly acquired in the same orphanage in Bamako, Mali, before the children were adopted by French families. An inappropriate use of ceftriaxone was probably the cause of the emergence of such strains. There is an urgent need to determine the origin of the contamination and to introduce adequate antibiotic protocols into this orphanage to prevent further transmission and dissemination. Screening for infections and follow-up, adapted to the origin of the internationally adopted children, should be recommended. PMID:15184415

  13. Effects of norspermidine and spermidine on biofilm formation by potentially pathogenic Escherichia coli and Salmonella enterica wild-type strains.

    PubMed

    Nesse, Live L; Berg, Kristin; Vestby, Lene K

    2015-03-01

    Polyamines are present in all living cells. In bacteria, polyamines are involved in a variety of functions, including biofilm formation, thus indicating that polyamines may have potential in the control of unwanted biofilm. In the present study, the effects of the polyamines norspermidine and spermidine on biofilms of 10 potentially pathogenic wild-type strains of Escherichia coli serotype O103:H2, Salmonella enterica subsp. enterica serovar Typhimurium, and S. enterica serovar Agona were investigated. We found that exogenously supplied norspermidine and spermidine did not mediate disassembly of preformed biofilm of any of the E. coli and S. enterica strains. However, the polyamines did affect biofilm production. Interestingly, the two species reacted differently to the polyamines. Both polyamines reduced the amount of biofilm formed by E. coli but tended to increase biofilm formation by S. enterica. Whether the effects observed were due to the polyamines specifically targeting biofilm formation, being toxic for the cells, or maybe a combination of the two, is not known. However, there were no indications that the effect was mediated through binding to exopolysaccharides, as earlier suggested for E. coli. Our results indicate that norspermidine and spermidine do not have potential as inhibitors of S. enterica biofilm. Furthermore, we found that the commercial polyamines used contributed to the higher pH of the test medium. Failure to acknowledge and control this important phenomenon may lead to misinterpretation of the results. PMID:25595767

  14. Classification of Salmonella enterica serotypes with selective bands using visible/NIR hyperspectral microscope images.

    PubMed

    Eady, M; Park, B

    2016-07-01

    Optical detection of foodborne bacteria such as Salmonella classifies bacteria by analysing spectral data, and has potential for rapid detection. In this experiment hyperspectral microscopy is explored as a means for classifying five Salmonella serotypes. Initially, the microscope collects 89 spectral measurements between 450 and 800 nm. Here, the objective was to develop correct classification of five serotypes with optimal spectral bands selected through multivariate data analysis (MVDA), thus reducing the data processing and storage requirement necessary for practical application in the food industry. An upright digital microscope is equipped with an acousto-optical tuneable filter, electron multiplying charge-coupled device, and metal halide lighting source. Images for each of the five serotypes were collected, and informative bands were identified through a principal component analysis, for four abbreviated spectral ranges containing 3, 7, 12 and 20 spectral bands. The experiment was repeated with an independent repetition and images were collected at each of the reduced band sets, identified by the first repetition. A support vector machine (SVM) was used to classify serotypes. Results showed that with the first repetition, classification accuracy decreased from 99.5% (89 bands) to 84.5% (3 bands), whereas the second repetition showed classification accuracies of 100%, possibly due to a reduction in spectral noise. The support vector machine regression (SVMR) was applied with cross-validation, and had R(2) calibration and validation values >0.922. Although classification accuracies through SVM classification showed that as little as 3 bands were able to classify 100% of the samples, the SVMR shows that the smallest root-mean squared-error values were 0.001 and 0.002 for 20 and 12 bands, respectively, suggesting that the 12 band range collected between 586 and 630 nm is optimal for classifying bacterial serotypes, with only the informative HMI bands selected

  15. Population Dynamics of Salmonella enterica Serotypes in Commercial Egg and Poultry Production ▿

    PubMed Central

    Foley, Steven L.; Nayak, Rajesh; Hanning, Irene B.; Johnson, Timothy J.; Han, Jing; Ricke, Steven C.

    2011-01-01

    Fresh and processed poultry have been frequently implicated in cases of human salmonellosis. Furthermore, increased consumption of meat and poultry has increased the potential for exposure to Salmonella enterica. While advances have been made in reducing the prevalence and frequency of Salmonella contamination in processed poultry, there is mounting pressure on commercial growers to prevent and/or eliminate these human pathogens in preharvest production facilities. Several factors contribute to Salmonella colonization in commercial poultry, including the serovar and the infectious dose. In the early 1900s, Salmonella enterica serovars Pullorum and Gallinarum caused widespread diseases in poultry, but vaccination and other voluntary programs helped eradicate pullorum disease and fowl typhoid from commercial flocks. However, the niche created by the eradication of these serovars was likely filled by S. Enteritidis, which proliferated in the bird populations. While this pathogen remains a significant problem in commercial egg and poultry production, its prevalence among poultry has been declining since the 1990s. Coinciding with the decrease of S. Enteritidis, S. Heidelberg and S. Kentucky have emerged as the predominant serovars in commercial broilers. In this review, we have highlighted bacterial genetic and host-related factors that may contribute to such shifts in Salmonella populations in commercial poultry and intervention strategies that could limit their colonization. PMID:21571882

  16. Attribution of Salmonella enterica serotype Hadar infections using antimicrobial resistance data from two points in the food supply system.

    PubMed

    Vieira, A R; Grass, J; Fedorka-Cray, P J; Plumblee, J R; Tate, H; Cole, D J

    2016-07-01

    A challenge to the development of foodborne illness prevention measures is determining the sources of enteric illness. Microbial subtyping source-attribution models attribute illnesses to various sources, requiring data characterizing bacterial isolate subtypes collected from human and food sources. We evaluated the use of antimicrobial resistance data on isolates of Salmonella enterica serotype Hadar, collected from ill humans, food animals, and from retail meats, in two microbial subtyping attribution models. We also compared model results when either antimicrobial resistance or pulsed-field gel electrophoresis (PFGE) patterns were used to subtype isolates. Depending on the subtyping model used, 68-96% of the human infections were attributed to meat and poultry food products. All models yielded similar outcomes, with 86% [95% confidence interval (CI) 80-91] to 91% (95% CI 88-96) of the attributable infections attributed to turkey, and 6% (95% CI 2-10) to 14% (95% CI 8-20) to chicken. Few illnesses (<3%) were attributed to cattle or swine. Results were similar whether the isolates were obtained from food animals during processing or from retail meat products. Our results support the view that microbial subtyping models are a flexible and robust approach for attributing Salmonella Hadar. PMID:26838291

  17. Survival of Salmonella enterica serotype Tennessee during simulated gastric passage is improved by low water activity and high fat content.

    PubMed

    Aviles, Bryan; Klotz, Courtney; Smith, Twyla; Williams, Robert; Ponder, Monica

    2013-02-01

    The low water activity (a(w) 0.3) of peanut butter prohibits the growth of Salmonella in a product; however, illnesses are reported from peanut butter contaminated with very small doses, suggesting the food matrix itself influences the infectious dose of Salmonella, potentially by improving Salmonella's survival in the gastrointestinal tract. The purpose of our study was to quantify the survival of a peanut butter outbreak-associated strain of Salmonella enterica serotype Tennessee when inoculated into peanut butters with different fat contents and a(w) (high fat, high a(w); high fat, low a(w); low fat, high a(w); low fat, low a(w)) and then challenged with a simulated gastrointestinal system. Exposures to increased fat content and decreased a(w) both were associated with a protective effect on the survival of Salmonella Tennessee in the simulated gastric fluid compared with control cells. After a simulated intestinal phase, the populations of Salmonella Tennessee in the control and low-fat formulations were not significantly different; however, a 2-log CFU/g increase occurred in high-fat formulations. This study demonstrates that cross-protection from low-a(w) stress and the presence of high fat results in improved survival in the low pH of the stomach. The potential for interaction of food matrix and stress adaptations could influence the virulence of Salmonella and should be considered for risk analysis. PMID:23433384

  18. A multistate outbreak of Salmonella enterica serotype typhimurium infection linked to raw milk consumption--Ohio, 2003.

    PubMed

    Mazurek, Jacek; Salehi, Ellen; Propes, Dennis; Holt, Jo; Bannerman, Tammy; Nicholson, Lisa M; Bundesen, Mark; Duffy, Rosemary; Moolenaar, Ronald L

    2004-10-01

    In December 2002, the Ohio Department of Health was notified of two children with Salmonella infection. Both had a history of drinking raw milk from a combination dairy-restaurant-petting zoo (dairy). The dairy was the only establishment in Ohio licensed to sell raw milk and reported 1.35 million visitors annually. We investigated to determine the extent of the outbreak and identify illness risk factors. A case patient was any person with pulsed-field gel electrophoresis-matched Salmonella enterica serotype Typhimurium from 30 November 2002 to 18 February 2003. Sixty-two met the confirmed case definition. Forty dairy case patient patrons were included in a case-control study; 56 controls were their well meal companions. Consumption of raw milk was found to be associated with illness (odds ratio, 45.1; 95% confidence interval, 8.8 to 311.9). The dairy discontinued selling raw milk. Because 27 other states still allow the sale of raw milk, awareness of the hazards of its consumption should be raised and relevant regulations carefully reviewed. PMID:15508625

  19. Genetically Similar Isolates of Salmonella enterica Serotype Enteritidis Persistent in China for a Long-Term Period.

    PubMed

    Song, Qifa; Shen, Xuanyi; Yang, Yuanbin; Zhang, Danyang; Gao, Hong

    2016-07-01

    Salmonella enterica serotype Enteritidis (S. Enteritidis) is an important causative agent of nontyphoidal salmonellosis in human populations. In this study, we collected 72 S. Enteritidis strains from 2004 to 2014 in Ningbo, mid-east China. Of the 72 strains, we identified a dominant clone of 58 strains recovered from patient's feces (n = 48), blood (n = 1), pleural effusion (n = 1), chickens (n = 3), and dessert cakes (n = 5) by pulsed-field gel electrophoresis (PFGE) and variable-number of tandem repeat analysis (MLVA). The profile arrangements of MLVA were SE1-SE2-SE3-SE5-SE6-SE8-SE9: 4-4-3-11-10-1-3. These dominant strains were susceptible to ampicillin, chloramphenicol, tetracycline, ciprofloxacin, gentamicin, cefotaxime and trimethoprim-sulfamethoxazole, and resistant to nalidixic acid. Additionally, all isolates harboured virulence genes invA, sipA, sopE, and spvB when tested by PCR. Our results reveal that genetically similar S. Enteritidis strains which accounted for several outbreaks as well as blood infection and pleural cavity infection are prevalent in China for a long-term period. This situation calls for further attention in the prevention and control of foodborne disease caused by Salmonella species. PMID:27228342

  20. Identification and typing of Salmonella enterica serotypes isolated from guinea fowl (Numida meleagris) farms in Benin during four laying seasons (2007 to 2010).

    PubMed

    Boko, C Kadoéito; Kpodekon, T Marc; Duprez, Jean-Noël; Imberechts, Hein; Taminiau, Bernard; Bertrand, Sophie; Mainil, Jacques G

    2013-02-01

    The main problem for the local guinea fowl (Numida meleagris) traditional farming and raising system in north-east Benin is the high mortality rate of the keets (up to 70%) due to a combination of climatic, nutritional, hygienic and infectious causes. The present study was carried out to identify and compare the isolates of Salmonella enterica from necropsied keets, laying guinea fowl, surrogate hen mothers, other contact animal species and farmers during four laying seasons (2007 to 2010). S. enterica belonging to eight different serotypes (Adelaide, Farakan, Kingston, Legon, Luke, Oakland, Sangalkam and Teshie) and one untypable isolate were isolated from 13 to 19% of the necropsied keets. The serotypes Adelaide, Farakan, Luke, Sangalkam and Teshie and the untypable isolate were isolated in only one township during 1 year of sampling, while serotypes Oakland, Legon and Kingston were present in two to three townships for 2 to 3 years of sampling. Serotypes Farakan, Kingston, Legon, Oakland and Sangalkam were also isolated from faecal samples of laying guinea fowl and/or surrogate domestic fowl hen mothers. Further comparison by pulsed-field gel electrophoresis and virulotyping provided evidence for their clonality within each of those five serotypes and therefore for the adult guinea fowl and/or hens as the most probable origin of contamination of the keets. The antibiotic resistance profiles, with all isolates resistant to oxacillin, sulfamethoxazol and colistin, emphasize the rise of antibiotic resistance in salmonellas from guinea fowl in this area and the need for alternative therapy policies for these birds. PMID:23391175

  1. Salmonella enterica serotype Javiana infections associated with amphibian contact, Mississippi, 2001.

    PubMed Central

    Srikantiah, P.; Lay, J. C.; Hand, S.; Crump, J. A.; Campbell, J.; Van Duyne, M. S.; Bishop, R.; Middendor, R.; Currier, M.; Mead, P. S.; Mølbak, K.

    2004-01-01

    Salmonella Javiana is a Salmonella serotype that is restricted geographically in the United States to the Southeast. During the summer of 2001, the number of reported S. Javiana infections in Mississippi increased sevenfold. To identify sources of infection, we conducted a case-control study, defining a case as an infection with S. Javiana between August and September in a Mississippi resident. We enrolled 55 cases and 109 controls. Thirty (55%) case patients reported exposure to amphibians, defined as owning, touching, or seeing an amphibian on one's property, compared with 30 (29%) controls (matched odds ratio 2.8, P=0.006). Contact with amphibians and their environments may be a risk factor for human infection with S. Javiana. The geographic pattern of S. Javiana infections in the United States mimics the distribution of certain amphibian species in the Southeast. Public health officials should consider amphibians as potential sources of salmonellosis, and promote hand washing after contact with amphibians. PMID:15061502

  2. An Outbreak of Food-Borne Typhoid Fever Due to Salmonella enterica Serotype Typhi in Japan Reported for the First Time in 16 Years

    PubMed Central

    Kobayashi, Tetsuro; Kutsuna, Satoshi; Hayakawa, Kayoko; Kato, Yasuyuki; Ohmagari, Norio; Uryu, Hideko; Yamada, Ritsuko; Kashiwa, Naoyuki; Nei, Takahito; Ehara, Akihito; Takei, Reiko; Mori, Nobuaki; Yamada, Yasuhiro; Hayasaka, Tomomi; Kagawa, Narito; Sugawara, Momoko; Suzaki, Ai; Takahashi, Yuno; Nishiyama, Hiroyuki; Morita, Masatomo; Izumiya, Hidemasa; Ohnishi, Makoto

    2016-01-01

    For the first time in 16 years, a food-borne outbreak of typhoid fever due to Salmonella enterica serotype Typhi was reported in Japan. Seven patients consumed food in an Indian buffet at a restaurant in the center of Tokyo, while one was a Nepali chef in the restaurant, an asymptomatic carrier and the implicated source of this outbreak. The multiple-locus variable-number tandem repeat analysis showed 100% consistency in the genomic sequence for five of the eight cases. PMID:26621565

  3. Case Report: An Outbreak of Food-Borne Typhoid Fever Due to Salmonella enterica Serotype Typhi in Japan Reported for the First Time in 16 Years.

    PubMed

    Kobayashi, Tetsuro; Kutsuna, Satoshi; Hayakawa, Kayoko; Kato, Yasuyuki; Ohmagari, Norio; Uryu, Hideko; Yamada, Ritsuko; Kashiwa, Naoyuki; Nei, Takahito; Ehara, Akihito; Takei, Reiko; Mori, Nobuaki; Yamada, Yasuhiro; Hayasaka, Tomomi; Kagawa, Narito; Sugawara, Momoko; Suzaki, Ai; Takahashi, Yuno; Nishiyama, Hiroyuki; Morita, Masatomo; Izumiya, Hidemasa; Ohnishi, Makoto

    2016-02-01

    For the first time in 16 years, a food-borne outbreak of typhoid fever due to Salmonella enterica serotype Typhi was reported in Japan. Seven patients consumed food in an Indian buffet at a restaurant in the center of Tokyo, while one was a Nepali chef in the restaurant, an asymptomatic carrier and the implicated source of this outbreak. The multiple-locus variable-number tandem repeat analysis showed 100% consistency in the genomic sequence for five of the eight cases. PMID:26621565

  4. Whole Genome DNA Sequence Analysis of Salmonella subspecies enterica serotype Tennessee obtained from related peanut butter foodborne outbreaks.

    PubMed Central

    Wilson, Mark R.; Brown, Eric; Keys, Chris; Strain, Errol; Luo, Yan; Muruvanda, Tim; Grim, Christopher; Jean-Gilles Beaubrun, Junia; Jarvis, Karen; Ewing, Laura; Gopinath, Gopal; Hanes, Darcy; Allard, Marc W.; Musser, Steven

    2016-01-01

    Establishing an association between possible food sources and clinical isolates requires discriminating the suspected pathogen from an environmental background, and distinguishing it from other closely-related foodborne pathogens. We used whole genome sequencing (WGS) to Salmonella subspecies enterica serotype Tennessee (S. Tennessee) to describe genomic diversity across the serovar as well as among and within outbreak clades of strains associated with contaminated peanut butter. We analyzed 71 isolates of S. Tennessee from disparate food, environmental, and clinical sources and 2 other closely-related Salmonella serovars as outgroups (S. Kentucky and S. Cubana), which were also shot-gun sequenced. A whole genome single nucleotide polymorphism (SNP) analysis was performed using a maximum likelihood approach to infer phylogenetic relationships. Several monophyletic lineages of S. Tennessee with limited SNP variability were identified that recapitulated several food contamination events. S. Tennessee clades were separated from outgroup salmonellae by more than sixteen thousand SNPs. Intra-serovar diversity of S. Tennessee was small compared to the chosen outgroups (1,153 SNPs), suggesting recent divergence of some S. Tennessee clades. Analysis of all 1,153 SNPs structuring an S. Tennessee peanut butter outbreak cluster revealed that isolates from several food, plant, and clinical isolates were very closely related, as they had only a few SNP differences between them. SNP-based cluster analyses linked specific food sources to several clinical S. Tennessee strains isolated in separate contamination events. Environmental and clinical isolates had very similar whole genome sequences; no markers were found that could be used to discriminate between these sources. Finally, we identified SNPs within variable S. Tennessee genes that may be useful markers for the development of rapid surveillance and typing methods, potentially aiding in traceback efforts during future

  5. Nosocomial Outbreak of a Novel Extended-Spectrum β-Lactamase Salmonella enterica Serotype Isangi Among Surgical Patients.

    PubMed

    Suleyman, Geehan; Tibbetts, Robert; Perri, Mary Beth; Vager, Dora; Xin, Yuan; Reyes, Katherine; Samuel, Linoj; Chami, Eman; Starr, Patricia; Pietsch, Jennifer; Zervos, Marcus J; Alangaden, George

    2016-08-01

    OBJECTIVE Nosocomial outbreaks caused by Salmonella are rare. We describe the investigation and control of a cluster of novel extended-spectrum β-lactamase (ESBL) Salmonella enterica serotype Isangi in a hospital in southeastern Michigan. METHODS An epidemiologic investigation, including case-control study, assessment of infection control practices and environmental cultures, was performed to identify modes of transmission. Healthcare workers (HCWs) exposed to case patients were screened. Strain relatedness was determined using pulsed-field gel electrophoresis (PFGE); ESBL confirmation was conducted using real-time PCR. Control measures were implemented to prevent further transmission. RESULTS Between September 2 and October 22, 2015, 19 surgical patients, including 10 organ transplant recipients and 1 HCW, had positive S. Isangi cultures. Of these case patients and HCW, 13 had gastroenteritis, 2 had bacteremia, 1 had surgical-site infection, and 4 were asymptomatic. Pulsed-field gel electrophoresis (PFGE) showed 89.5% similarity among the isolates in these cases. Isolates with resistant-phenotypes possessed plasmid-mediated CTX-M15 ESBL. A total of 19 case patients were compared with 57 control participants. Case patients had significantly higher odds of exposure to an intraoperative transesophageal (TEE) probe (adjusted odds ratio 9.0; 95% confidence interval, 1.12-72.60; P=.02). Possible cross-transmission occurred in the HCW and 2 patients. Cultures of TEE probes and the environment were negative. The outbreak ended after removal of TEE probes, modification of reprocessing procedures, implementation of strict infection control practices, and enhanced environmental cleaning. CONCLUSIONS We report the first nosocomial ESBL S. Isangi outbreak in the United States. Multiple control measures were necessary to interrupt transmission of this gastrointestinal pathogen. Exposure to possibly contaminated TEE probes was associated with transmission. Periodic monitoring

  6. Biofilm formation of Salmonella serotypes in simulated meat processing environments and its relationship to cell characteristics.

    PubMed

    Wang, Huhu; Ding, Shijie; Dong, Yang; Ye, Keping; Xu, Xinglian; Zhou, Guanghong

    2013-10-01

    Salmonella attached to meat contact surfaces encountered in meat processing facilities may serve as a source of cross-contamination. In this study, the influence of serotypes and media on biofilm formation of Salmonella was investigated in a simulated meat processing environment, and the relationships between biofilm formation and cell characteristics were also determined. All six serotypes (Salmonella enterica serotype Heidelberg, Salmonella Derby, Salmonella Agona, Salmonella Indiana, Salmonella Infantis, and Salmonella Typhimurium) can readily form biofilms on stainless steel surfaces, and the amounts of biofilms were significantly influenced by the serotypes, incubation media, and incubation time used in this study. Significant differences in cell surface hydrophobicity, autoaggregation, motility, and growth kinetic parameters were observed between individual serotypes tested. Except for growth kinetic parameters, the cell characteristics were correlated with the ability of biofilm formation incubated in tryptic soy broth, whereas no correlation with biofilm formation incubated in meat thawing-loss broth (an actual meat substrate) was found. Salmonella grown in meat thawing-loss broth showed a "cloud-shaped" morphology in the mature biofilm, whereas when grown in tryptic soy broth it had a "reticulum-shaped" appearance. Our study provides some practical information to understand the process of biofilm formation on meat processing contact surfaces. PMID:24112581

  7. An outbreak of Salmonella enterica serotype Litchfield infection in Australia linked to consumption of contaminated papaya.

    PubMed

    Gibbs, Robyn; Pingault, Nevada; Mazzucchelli, Terry; O'Reilly, Lyn; MacKenzie, Brian; Green, Jennifer; Mogyorosy, Ray; Stafford, Russell; Bell, Robert; Hiley, Lester; Fullerton, Kathleen; Van Buynder, Paul

    2009-05-01

    An outbreak of 26 cases of Salmonella Litchfield infection occurred in the states of Western Australia and Queensland between October 2006 and January 2007. A case-control study was conducted with 12 cases and 24 controls, and a significant association was found between illness and consumption of papaya (odds ratio, 32.8; 95% confidence interval, 2.71 to 883.5). Papaya samples were collected from 26 stores in Western Australia, and 9 of 38 samples were contaminated with Salmonella Litchfield. These samples had pulsed-field gel electrophoresis patterns and multilocus variable-number tandem-repeat analysis profiles indistinguishable from the outbreak strain. Three farms in Western Australia supplied the contaminated papaya, and two of these farms were inspected. Salmonella Litchfield was not detected in papaya samples, fungal sprays, or water samples from the farms; however, at one farm other serotypes of Salmonella were detected in untreated river water that was used for washing papaya. Only treated potable water should be used for washing fresh produce that is to be eaten raw. PMID:19517740

  8. Relative survival of four serotypes of Salmonella enterica in low-water activity whey protein powder held at 36 and 70°C at various water activity levels

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Salmonella enterica is the leading cause of health burdens in the United States. Although the pathogen is not able to grow at aw levels below 0.94, it can survive in low-moisture foods for long periods of time. Temperature, aw, substrate and serotype affect its persistence. The aim of this study was...

  9. Draft Genome Sequences of Two Salmonella enterica Serotype Infantis Strains Isolated from a Captive Western Lowland Gorilla (Gorilla gorilla gorilla) and a Cohabitant Black and White Tegu (Tupinambis merianae) in Brazil.

    PubMed

    Paixão, Tatiane A; Coura, Fernanda M; Malta, Marcelo C C; Tinoco, Herlandes P; Pessanha, Angela T; Pereira, Felipe L; Leal, Carlos A G; Heinemann, Marcos B; Figueiredo, Henrique C P; Santos, Renato L

    2016-01-01

    The draft genome sequences of two Salmonella enterica serotype Infantis isolates are reported here. One of the strains was isolated from a western lowland gorilla (Gorilla gorilla gorilla) with colitis. The second strain was isolated from a reptile that inhabited the same premises. Whole-genome sequencing demonstrated that these isolates were not clonal. PMID:26798099

  10. Draft Genome Sequences of Two Salmonella enterica Serotype Infantis Strains Isolated from a Captive Western Lowland Gorilla (Gorilla gorilla gorilla) and a Cohabitant Black and White Tegu (Tupinambis merianae) in Brazil

    PubMed Central

    Paixão, Tatiane A.; Coura, Fernanda M.; Malta, Marcelo C. C.; Tinoco, Herlandes P.; Pessanha, Angela T.; Pereira, Felipe L.; Leal, Carlos A. G.; Heinemann, Marcos B.; Figueiredo, Henrique C. P.

    2016-01-01

    The draft genome sequences of two Salmonella enterica serotype Infantis isolates are reported here. One of the strains was isolated from a western lowland gorilla (Gorilla gorilla gorilla) with colitis. The second strain was isolated from a reptile that inhabited the same premises. Whole-genome sequencing demonstrated that these isolates were not clonal. PMID:26798099

  11. Analysis of pSC138, the multidrug resistance plasmid of Salmonella enterica serotype Choleraesuis SC-B67.

    PubMed

    Ye, Jiehua; Su, Lin-Hui; Chen, Chyi-Liang; Hu, Songnian; Wang, Jianbing; Yu, Jun; Chiu, Cheng-Hsun

    2011-03-01

    Salmonella enterica serotype Choleraesuis (S. Choleraesuis) usually causes systemic infections in man and needs antimicrobial treatment. Multidrug resistance (MDR) in S. Choleraesuis is thus a great concern in the treatment of systemic non-typhoid salmonellosis. A large plasmid, pSC138, was identified in 2002 from a S. Choleraesuis strain SC-B67 that was resistant to all antimicrobial agents commonly used to treat salmonellosis, including ciprofloxacin and ceftriaxone. Complete DNA sequence of the plasmid had been determined previously (Chiu et al., 2005). In the present study, the sequence of pSC138 was reannotated in detail and compared with several newly sequenced plasmids. Some transposable elements and drug resistance genes were further delineated. Plasmid pSC138 was 138,742 bp in length and consisted of 177 open reading frames (ORFs). While 134 of the ORFs displayed significant identity levels to other plasmid and prokaryotic sequences, the remaining 43 ORFs have not been previously reported. Mobile elements, including two integrons, seven insertion sequences and eight transposons, and a truncated prophage together encompass at least 66,781 bp (48.1%) of the plasmid genome. The sequence of pSC138 consists of three major regions: a large composite transposable region Tn6088 with a Tn21-like backbone inserted by a variety of integrons or transposable elements; a transfer/maintenance region that contains a conserved ISEcp1-mediated transposon-like element Tn6092, carrying an AmpC gene, bla(CMY-2), that confers the ceftriaxone resistance; and a Rep_3 type of replication region. Another seven bacteremic strains of S. Choleraesuis that expressed the same MDR phenotype were identified during 2003-2008. The same Rep_3 type replicase and the bla(CMY-2)-containing, ISEcp1-mediated transposon-like element were found in the MDR isolates, suggesting a successful preservation and dissemination of the MDR plasmid. Comparison of pSC138 with other recently published plasmids

  12. Reduction of Salmonella enterica serotype Poona and background microbiota on fresh-cut cantaloupe by electron beam irradiation.

    PubMed

    Palekar, Mangesh P; Taylor, T Matthew; Maxim, Joseph E; Castillo, Alejandro

    2015-06-01

    The efficacy of electron beam (e-beam) irradiation processing to reduce Salmonella enterica serotype Poona on surfaces of fresh-cut cantaloupe, and the impact of e-beam irradiation processing on the numbers of indigenous microorganisms were determined. Additionally, the D10-value for S. Poona reduction on the cut cantaloupe was also determined. Fresh-cut cantaloupe pieces, inoculated with S. Poona to 7.8 log10 CFU/g, were exposed to 0.0, 0.7, or 1.5 kGy. Surviving S. Poona, lactic acid bacteria (LAB), and fungi (yeasts, molds) were periodically enumerated on appropriate media over 21 days of storage at 5 °C. Cantaloupe surface pH was measured for irradiated cantaloupe across the 21 day storage period. To determine the D10-value of S. Poona, cantaloupe discs were inoculated and exposed to increasing radiation dosages between 0 and 1.06 kGy; surviving pathogen cells were selectively enumerated. S. Poona was significantly reduced by irradiation; immediate reductions following exposure to 0.7 and 1.5 kGy were 1.1 and 3.6 log10 CFU/g, respectively. After 21 days, S. Poona numbers were between 4.0 and 5.0 log10 CFU/g less than untreated samples at zero-time. Yeasts were not reduced significantly (p ≥ 0.05) by e-beam irradiation and grew slowly but steadily during storage. Counts of LAB and molds were initially reduced with 1.5 kGy (p<0.05) but then LAB recovered grew to high numbers, whereas molds slowly declined for irradiated and control samples. Cantaloupe pH declined during storage, with the greatest decrease in untreated control cantaloupe (p<0.05). The D10-value for S. Poona was determined to be 0.211 kGy, and this difference from the reductions observed in the cut cantaloupe studies may be due to the more precise dose distribution obtained in the thin and flat cantaloupe pieces used for the D10-value experiments. The effect of e-beam irradiation at the same doses used in this study was determined in previous studies to have no negative effect in the quality of

  13. Detection of Salmonella enterica in Magellanic penguins (Spheniscus magellanicus) of Chilean Patagonia: evidences of inter-species transmission.

    PubMed

    Dougnac, C; Pardo, C; Meza, K; Arredondo, C; Blank, O; Abalos, P; Vidal, R; Fernandez, A; Fredes, F; Retamal, P

    2015-04-01

    Patagonia in southern South America is among the few world regions where direct human impact is still limited but progressively increasing, mainly represented by tourism, farming, fishing and mining activities. The sanitary condition of Patagonian wildlife is unknown, in spite of being critical for the assessment of anthropogenic effects there. The aim of this study was the characterization of Salmonella enterica strains isolated from wild colonies of Magellanic penguins (Spheniscus magellanicus) located in Magdalena Island and Otway Sound, in Chilean Patagonia. Eight isolates of Salmonella were found, belonging to Agona and Enteritidis serotypes, with an infection rate of 0·38%. Resistance to ampicillin, cefotaxime, ceftiofur and tetracycline antimicrobials were detected, and some of these strains showed genotypic similarity with Salmonella strains isolated from humans and gulls, suggesting inter-species transmission cycles and strengthening the role of penguins as sanitary sentinels in the Patagonian ecosystem. PMID:25148565

  14. Complete Genome and Methylome Sequences of Salmonella enterica subsp. enterica Serovar Panama (ATCC 7378) and Salmonella enterica subsp. enterica Serovar Sloterdijk (ATCC 15791).

    PubMed

    Yao, Kuan; Muruvanda, Tim; Roberts, Richard J; Payne, Justin; Allard, Marc W; Hoffmann, Maria

    2016-01-01

    Salmonella enterica spp. are pathogenic bacteria commonly associated with food-borne outbreaks in human and animals. Salmonella enterica spp. are characterized into more than 2,500 different serotypes, which makes epidemiological surveillance and outbreak control more difficult. In this report, we announce the first complete genome and methylome sequences from two Salmonella type strains associated with food-borne outbreaks, Salmonella enterica subsp. enterica serovar Panama (ATCC 7378) and Salmonella enterica subsp. enterica serovar Sloterdijk (ATCC 15791). PMID:26988049

  15. Complete Genome and Methylome Sequences of Salmonella enterica subsp. enterica Serovar Panama (ATCC 7378) and Salmonella enterica subsp. enterica Serovar Sloterdijk (ATCC 15791)

    PubMed Central

    Yao, Kuan; Muruvanda, Tim; Roberts, Richard J.; Payne, Justin; Allard, Marc W.

    2016-01-01

    Salmonella enterica spp. are pathogenic bacteria commonly associated with food-borne outbreaks in human and animals. Salmonella enterica spp. are characterized into more than 2,500 different serotypes, which makes epidemiological surveillance and outbreak control more difficult. In this report, we announce the first complete genome and methylome sequences from two Salmonella type strains associated with food-borne outbreaks, Salmonella enterica subsp. enterica serovar Panama (ATCC 7378) and Salmonella enterica subsp. enterica serovar Sloterdijk (ATCC 15791). PMID:26988049

  16. Antimicrobial resistance and extended-spectrum β-lactamases of Salmonella enterica serotypes isolated from livestock and processed food in Portugal: an update.

    PubMed

    Figueiredo, Rui; Henriques, Ana; Sereno, Rui; Mendonça, Nuno; da Silva, Gabriela Jorge

    2015-02-01

    As Salmonella is a common foodborne pathogen, the present study aimed to determine the distribution of Salmonella enterica serotypes isolated during 2011-2012 from poultry, swine, cattle, and processed food in Portugal, and to characterize the antimicrobial susceptibility and the extended-spectrum β-lactamases (ESBLs). Results were also compared with data obtained before the implementation of the National Control Program in Poultry and the ban of antimicrobial agents in animal feed in the European Union (EU). A total of 14 serotypes were identified, from 258 isolates recovered, with Salmonella Typhimurium (32.6%, n=84) and Salmonella Enteritidis (10.1%, n=26) being the most common. Salmonella Enteritidis in poultry was less frequent than in previous studies, which might be associated with the implementation of the National Control Program for Salmonella in poultry. Nevertheless, other serotypes seem to occupy this biological niche, and may be more common in human salmonellosis in the future. The majority of isolates (70.2%, n=181) were resistant to at least one class of antimicrobial agent and exhibited higher frequency of resistance to tetracycline (47.7%, n=123) and ampicillin (36.0%, n=93), with Salmonella Typhimurium being the more resistant serotype. Resistance to fluoroquinolones was shown in 8% (n=21) of isolates, a lower value compared to data obtained before 2004. ESBLs producers Salmonella Typhimurium bla(CTX-M-1) and Salmonella Enteritidis bla(SHV-12) were isolated from swine and poultry, respectively. The bla(CTX-M-1) and bla(SHV-12) genes were carried on conjugative plasmids of IncHI2replicon types and IncI1, respectively. This was the first report of a bla(CTX-M-1) in Salmonella Typhimurium in Portugal. Overall, the results revealed changes in animal origin Salmonella serotypes, mainly emerging serotypes, in frequency of resistance, and in occurrence of ESBLs-producing Salmonella. The control measures taken by the EU seem to have some impact on the

  17. The characterization of Salmonella enterica serotypes isolated from the scalder tank water of a commercial poultry processing plant: Recovery of a multidrug-resistant Heidelberg strain.

    PubMed

    Rothrock, Michael J; Ingram, Kimberly D; Gamble, John; Guard, Jean; Cicconi-Hogan, Kellie M; Hinton, Arthur; Hiett, Kelli L

    2015-03-01

    The recent multistate outbreak of a multidrug-resistant (MDR) Salmonella Heidelberg strain from commercial poultry production highlights the need to better understand the reservoirs of these zoonotic pathogens within the commercial poultry production and processing environment. As part of a larger study looking at temporal changes in microbial communities within the major water tanks within a commercial processing facility, this paper identifies and characterizes Salmonella enterica isolated from the water in a final scalder tank at 3 times during a typical processing day: prior to the birds entering the tank (start), halfway through the processing day (mid), and after the final birds were scalded (end). Over 3 consecutive processing days, no Salmonella were recovered from start-of-day water samples, while a total of 56 Salmonella isolates were recovered from the mid-day and end-of-day scalder water samples. Traditional and newer PCR-based serotyping methods eventually identified these isolates as either group C3 S. Kentucky (n=45) and group B S. Heidelberg (n=11). While none of the S. Kentucky isolates possessed any resistances to the antimicrobials tested, all S. Heidelberg isolates were found to be multidrug resistant to 5 specific antimicrobials representing 3 antimicrobial classes. Due to the potential public health impact of S. Heidelberg and the recent nationwide poultry-associated outbreak of multidrug-resistant S. Heidelberg, future studies should focus on understanding the transmission and environmental growth dynamics of this serotype within the commercial poultry processing plant environment. PMID:25681479

  18. Intergenic Sequence Ribotyping using a region neighboring dkgB links genovar to Kauffman-White serotype of Salmonella enterica

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Thirty six (36) unique sequences which varied in length from 258bp to 530bp were found for Salmonella enterica strains and isolates that are not present in public databases following BLAST analysis searches for similarity. The sequences were found by application of Intergenic Sequence Ribotyping (IS...

  19. Characterization of blaCMY plasmids and their possible role in source attribution of salmonella enterica serotype typhimurium infections

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Salmonella is an important cause of foodborne illness; however, identifying the source of these infections can be difficult. This is especially true for Salmonella serotype Typhimurium, which is found in diverse agricultural niches. Cephalosporins are one of the primary treatment choices for complic...

  20. SEROTYPES AND ANTIMICROBIAL RESISTANCE OF SALMONELLA ENTERICA ISOLATED FROM PORK, CHICKEN MEAT AND LETTUCE, BANGKOK AND CENTRAL THAILAND.

    PubMed

    Niyomdecha, Nattamon; Mungkornkaew, Narissara; Samosornsuk, Worada

    2016-01-01

    Food of animal origins, particularly pork and chicken meat, has long been recognized as major sources of human salmonellosis. There have been recent reports of human salmonellosis outbreaks due to consumption of leafy green vegetables such as lettuce. In this study, 120 (40 pork, 40 chicken meat and 40 lettuce) samples were randomly collected from retail markets in Bangkok and central Thailand during June to August 2015 for Salmonella serotype identification and antimicrobial susceptibility testing. Salmonella was found in 82%, 62% and 20% of pork, chicken meat and lettuce samples, respectively. The top 5 most common Salmonella serotypes were Panama (15%), Schwarzengrund (12%), Rissen, Anatum, and Stanley (11% each), Albany (9%), and Indiana (8%). A high percentage of Salmonella isolated from food of animal origin were resistant to multiple antimicrobial drugs, including ampicillin, chloramphenicol, nalidixic acid, sulfamethoxazole-trimethoprim, and tetracycline. From antibiogram pattern analysis, the most common serotypes constituted isolates that were multidrug resistant. The study indicates that Salmonella was still present in various kinds of food and that certain serotypes have become predominant, a phenomenon not previously reported in Thailand. PMID:27086423

  1. Fast DNA Serotyping and Antimicrobial Resistance Gene Determination of Salmonella enterica with an Oligonucleotide Microarray-Based Assay

    PubMed Central

    Braun, Sascha D.; Ziegler, Albrecht; Methner, Ulrich; Slickers, Peter; Keiling, Silke; Monecke, Stefan; Ehricht, Ralf

    2012-01-01

    Salmonellosis caused by Salmonella (S.) belongs to the most prevalent food-borne zoonotic diseases throughout the world. Therefore, serotype identification for all culture-confirmed cases of Salmonella infection is important for epidemiological purposes. As a standard, the traditional culture method (ISO 6579:2002) is used to identify Salmonella. Classical serotyping takes 4–5 days to be completed, it is labor-intensive, expensive and more than 250 non-standardized sera are necessary to characterize more than 2,500 Salmonella serovars currently known. These technical difficulties could be overcome with modern molecular methods. We developed a microarray based serogenotyping assay for the most prevalent Salmonella serovars in Europe and North America. The current assay version could theoretically discriminate 28 O-antigens and 86 H-antigens. Additionally, we included 77 targets analyzing antimicrobial resistance genes. The Salmonella assay was evaluated with a set of 168 reference strains representing 132 serovars previously serotyped by conventional agglutination through various reference centers. 117 of 132 (81%) tested serovars showed an unique microarray pattern. 15 of 132 serovars generated a pattern which was shared by multiple serovars (e.g., S. ser. Enteritidis and S. ser. Nitra). These shared patterns mainly resulted from the high similarity of the genotypes of serogroup A and D1. Using patterns of the known reference strains, a database was build which represents the basis of a new PatternMatch software that can serotype unknown Salmonella isolates automatically. After assay verification, the Salmonella serogenotyping assay was used to identify a field panel of 105 Salmonella isolates. All were identified as Salmonella and 93 of 105 isolates (88.6%) were typed in full concordance with conventional serotyping. This microarray based assay is a powerful tool for serogenotyping. PMID:23056321

  2. Fast DNA serotyping and antimicrobial resistance gene determination of salmonella enterica with an oligonucleotide microarray-based assay.

    PubMed

    Braun, Sascha D; Ziegler, Albrecht; Methner, Ulrich; Slickers, Peter; Keiling, Silke; Monecke, Stefan; Ehricht, Ralf

    2012-01-01

    Salmonellosis caused by Salmonella (S.) belongs to the most prevalent food-borne zoonotic diseases throughout the world. Therefore, serotype identification for all culture-confirmed cases of Salmonella infection is important for epidemiological purposes. As a standard, the traditional culture method (ISO 6579:2002) is used to identify Salmonella. Classical serotyping takes 4-5 days to be completed, it is labor-intensive, expensive and more than 250 non-standardized sera are necessary to characterize more than 2,500 Salmonella serovars currently known. These technical difficulties could be overcome with modern molecular methods. We developed a microarray based serogenotyping assay for the most prevalent Salmonella serovars in Europe and North America. The current assay version could theoretically discriminate 28 O-antigens and 86 H-antigens. Additionally, we included 77 targets analyzing antimicrobial resistance genes. The Salmonella assay was evaluated with a set of 168 reference strains representing 132 serovars previously serotyped by conventional agglutination through various reference centers. 117 of 132 (81%) tested serovars showed an unique microarray pattern. 15 of 132 serovars generated a pattern which was shared by multiple serovars (e.g., S. ser. Enteritidis and S. ser. Nitra). These shared patterns mainly resulted from the high similarity of the genotypes of serogroup A and D1. Using patterns of the known reference strains, a database was build which represents the basis of a new PatternMatch software that can serotype unknown Salmonella isolates automatically. After assay verification, the Salmonella serogenotyping assay was used to identify a field panel of 105 Salmonella isolates. All were identified as Salmonella and 93 of 105 isolates (88.6%) were typed in full concordance with conventional serotyping. This microarray based assay is a powerful tool for serogenotyping. PMID:23056321

  3. Chromosomal Integration of the Extended-Spectrum β-Lactamase Gene blaCTX-M-15 in Salmonella enterica Serotype Concord Isolates from Internationally Adopted Children▿

    PubMed Central

    Fabre, Laëtitia; Delauné, Aurélia; Espié, Emmanuelle; Nygard, Karin; Pardos, Maria; Polomack, Lucette; Guesnier, Françoise; Galimand, Marc; Lassen, Jørgen; Weill, François-Xavier

    2009-01-01

    We report the emergence of Salmonella enterica isolates of serotype Concord (and its monophasic variant 6,7:l,v:-) producing the extended-spectrum β-lactamases (ESBLs) SHV-12 and CTX-M-15 in France and Norway between 2001 and 2006 (43 in France and 26 in Norway). The majority of these isolates were from adopted children from Ethiopia, most of whom were healthy carriers. Several symptomatic secondary cases were found in the adoptive families and health care facilities in France. Serotype Concord isolates collected before 2003 produced SHV-12 encoded on a 340-kb conjugative plasmid of replicon IncI1. Isolates collected after 2003 produced CTX-M-15. We detected two conjugative plasmids carrying blaCTX-M-15. One plasmid, approximately 300 kb in size, was positive for the IncHI2 replicon and the plasmid-mediated quinolone resistance gene qnrA1. The other plasmid, from one of the earliest CTX-M-15-producing isolates collected, was a fusion plasmid with IncY and IncA/C2 replicons and was 200 kb in size. However, we showed, using Southern hybridization of I-CeuI-digested chromosomal DNA and S1 nuclease analysis of plasmid DNA, that most isolates had a blaCTX-M-15 gene located on chromosomal DNA. Analysis of the flanking regions of the chromosomally located blaCTX-M-15 gene by cloning revealed an ISEcp1 truncated by an intact IS26 upstream from the blaCTX-M-15 gene and a truncated orf477 gene downstream from blaCTX-M-15. We found regions beyond the IS26 and the orf477 genes that were derived from IncA/C2 plasmids, suggesting the chromosomal integration of part of the blaCTX-M-15-carrying IncY and IncA/C2 fusion plasmid from early CTX-M-15-producing isolates. PMID:19273688

  4. Use of random forest to estimate population attributable fractions from a case-control study of Salmonella enterica serotype Enteritidis infections.

    PubMed

    Gu, W; Vieira, A R; Hoekstra, R M; Griffin, P M; Cole, D

    2015-10-01

    To design effective food safety programmes we need to estimate how many sporadic foodborne illnesses are caused by specific food sources based on case-control studies. Logistic regression has substantive limitations for analysing structured questionnaire data with numerous exposures and missing values. We adapted random forest to analyse data of a case-control study of Salmonella enterica serotype Enteritidis illness for source attribution. For estimation of summary population attributable fractions (PAFs) of exposures grouped into transmission routes, we devised a counterfactual estimator to predict reductions in illness associated with removing grouped exposures. For the purpose of comparison, we fitted the data using logistic regression models with stepwise forward and backward variable selection. Our results show that the forward and backward variable selection of logistic regression models were not consistent for parameter estimation, with different significant exposures identified. By contrast, the random forest model produced estimated PAFs of grouped exposures consistent in rank order with results obtained from outbreak data, with egg-related exposures having the highest estimated PAF (22·1%, 95% confidence interval 8·5-31·8). Random forest might be structurally more coherent and efficient than logistic regression models for attributing Salmonella illnesses to sources involving many causal pathways. PMID:25672399

  5. Multiple-locus variable number of tandem repeats analysis of Salmonella enterica serotype paratyphi A from Yuxi and comparison with isolates from the Chinese Medical Culture Collection Center

    PubMed Central

    YAO, YINGBO; CUI, XIAOYAN; CHEN, QINGSHAN; HUANG, XINRONG; ELMORE, BRADLEY; PAN, QING; WANG, SHUKUN; LIU, JIE

    2014-01-01

    The aim of the present study was to genotype Salmonella enterica serotype paratyphi A (SPA) isolated from Yuxi, China, in a multiple-locus variable number of tandem repeats (VNTRs) analysis (MLVA) and to compare them with isolates from the Chinese Medical Culture Collection Center (CMCC). Potential VNTRs were screened from the genomes of ATCC9150 and AKU_12601 using the Tandem Repeats Finder program. Nine VNTRs were established for MLVA typing of 195 SPA isolates from Yuxi and 20 isolates from CMCC. The dendogram for MLVA profiles and minimum spanning tree (MST) were drawn using the categorical coefficient calculated by BioNumerics software. A total of 23 MLVA types were identified in 215 SPA isolates and were grouped into six distinct cluster groups A, B, C, D, E and F. A total of 195 Yuxi SPA isolates were exclusively grouped into cluster C with nine MLVA genotypes. A total of 20 CMCC isolates were grouped in clusters A B, D, E and F with the other 14 MLVA types. The MLVA with nine VNTR loci, which was exploited in the present study, represents a successful strategy for genotyping SPA. Furthermore, the 195 Yuxi isolates appear to be closely related to each other and distinct from the 20 CMCC strains. PMID:24788795

  6. Development of a Loop Mediated Isothermal Amplification (LAMP) - Surface Enhanced Raman spectroscopy (SERS) Assay for the Detection of Salmonella Enterica Serotype Enteritidis

    PubMed Central

    Draz, Mohamed Shehata; Lu, Xiaonan

    2016-01-01

    As a major foodborne pathogen, Salmonella enterica serotype Enteritidis is increasingly rising as a global health concern. Here, we developed an integrated assay that combines loop mediated isothermal amplification (LAMP) and surface enhanced Raman spectroscopy (SERS) for DNA detection of S. Enteritidis using specifically designed Raman active Au-nanoprobes. The target DNA was amplified by LAMP and then labeled with Au-nanoprobes comprised of gold nanoparticle-modified with specific cy5/DNA probes to allow the detection by SERS. The sensitivity of the developed LAMP-SERS detection assay (66 CFU/mL) was ~100-fold higher than the conventional polymerase chain reaction (PCR) method. Significantly, this technique allowed highly specific detection of the target DNA of S. Enteritidis and could differentiate it from the DNA of closely related bacterial species or non-specific contamination, making it more accurate and reliable than the standard LAMP technique. The applicability of detection of S. Enteritidis in milk samples using LAMP-SERS assay was validated as well. In sum, the developed LAMP-SERS assay is highly specific and sensitive, and has the potential to be applied for rapid detection of different foodborne pathogens and other microbial contaminants. PMID:26941845

  7. Immediate Reduction of Salmonella enterica Serotype Typhimurium Viability via Membrane Destabilization following Exposure to Multiple-Hurdle Treatments with Heated, Acidified Organic Acid Salt Solutions▿†

    PubMed Central

    Milillo, S. R.; Martin, E.; Muthaiyan, A.; Ricke, S. C.

    2011-01-01

    The antimicrobial activity of organic acids in combination with nonchemical treatments was evaluated for inactivation of Salmonella enterica serotype Typhimurium within 1 min. It was observed that the effectiveness of the multiple-hurdle treatments was temperature (P ≤ 0.05) and pH (P ≤ 0.05) dependent and corresponded to the degree of organic acid lipophilicity (sodium acetate being least effective and sodium propionate being the most effective). This led to the hypothesis that the loss in viability was due at least in part to cell membrane disruption. Evaluation of osmotic response, potassium ion leakage, and transmission electron micrographs confirmed treatment effects on the cell membrane. Interestingly, all treatments, even those with no effect on viability, such as with sodium acetate, resulted in measurable cellular stress. Microarray experiments explored the specific response of S. Typhimurium to sodium acetate and sodium propionate, the most similar of the tested treatments in terms of pKa and ionic strength, and found little difference in the changes in gene expression following exposure to either, despite their very different effects on viability. Taken together, the results reported support our hypothesis that treatment with heated, acidified, organic acid salt solutions for 1 min causes loss of S. Typhimurium viability at least in part by membrane damage and that the degree of effectiveness can be correlated with lipophilicity of the organic acid. Overall, the data presented here indicate that a combined thermal, acidified sodium propionate treatment can provide an effective antimicrobial treatment against Salmonella. PMID:21478311

  8. Development of a Loop Mediated Isothermal Amplification (LAMP) - Surface Enhanced Raman spectroscopy (SERS) Assay for the Detection of Salmonella Enterica Serotype Enteritidis.

    PubMed

    Draz, Mohamed Shehata; Lu, Xiaonan

    2016-01-01

    As a major foodborne pathogen, Salmonella enterica serotype Enteritidis is increasingly rising as a global health concern. Here, we developed an integrated assay that combines loop mediated isothermal amplification (LAMP) and surface enhanced Raman spectroscopy (SERS) for DNA detection of S. Enteritidis using specifically designed Raman active Au-nanoprobes. The target DNA was amplified by LAMP and then labeled with Au-nanoprobes comprised of gold nanoparticle-modified with specific cy5/DNA probes to allow the detection by SERS. The sensitivity of the developed LAMP-SERS detection assay (66 CFU/mL) was ~100-fold higher than the conventional polymerase chain reaction (PCR) method. Significantly, this technique allowed highly specific detection of the target DNA of S. Enteritidis and could differentiate it from the DNA of closely related bacterial species or non-specific contamination, making it more accurate and reliable than the standard LAMP technique. The applicability of detection of S. Enteritidis in milk samples using LAMP-SERS assay was validated as well. In sum, the developed LAMP-SERS assay is highly specific and sensitive, and has the potential to be applied for rapid detection of different foodborne pathogens and other microbial contaminants. PMID:26941845

  9. Evolution of genes on the Salmonella Virulence plasmid phylogeny revealed from sequencing of the virulence plasmids of S. enterica serotype Dublin and comparative analysis.

    PubMed

    Chu, Chishih; Feng, Ye; Chien, An-Chi; Hu, Songnian; Chu, Chi-Hong; Chiu, Cheng-Hsun

    2008-11-01

    Salmonella enterica serotype Dublin harbors an approximately 80-kb virulence plasmid (pSDV), which mediates systemic infection in cattle. There are two types of pSDV: one is pSDVu (pOU1113) in strain OU7025 and the other pSDVr (pOU1115) in OU7409 (SD Lane) and many clinical isolates. Sequence analysis showed that pSDVr was a recombinant plasmid (co-integrate) of pSDVu and a plasmid similar to a 35-kb indigenous plasmid (pOU1114) of S. Dublin. Most of the F-transfer region in pSDVu was replaced by a DNA segment from the pOU1114-like plasmid containing an extra replicon and a pilX operon encoding for a type IV secretion system to form pSDVr. We reconstructed the particular evolutionary history of the seven virulence plasmids of Salmonella by comparative sequence analysis. The whole evolutionary process might begin with two different F-like plasmids (IncFI and IncFII), which then incorporated the spv operon and fimbriae operon from the chromosome to form the primitive virulence plasmids. Subsequently, these plasmids descended by deletion from a relatively large plasmid to smaller ones, with some recombination events occurring over time. Our results suggest that the phylogeny of virulence plasmids as a result of frequent recombination provides the opportunity for rapid evolution of Salmonella in response to the environmental cues. PMID:18718522

  10. Salmonella enterica serotype Typhimurium ShdA is an outer membrane fibronectin-binding protein that is expressed in the intestine.

    PubMed

    Kingsley, Robert A; Santos, Renato L; Keestra, A Marijke; Adams, L Garry; Bäumler, Andreas J

    2002-02-01

    The shdA gene is the only determinant known to be required for persistence of Salmonella enterica serotype Typhimurium (S. Typhimurium) in the murine caecum and for efficient and prolonged shedding of the organism with the faeces. To study the biological activity of the ShdA protein, we examined its expression and binding activity. ShdA was not detected with anti-ShdA antiserum in S. Typhimurium strain ATCC14028 grown in vitro, suggesting that this protein is not expressed under standard conditions of bacterial cultivation in the laboratory. However, in mice infected with S. Typhimurium, an immunofluorescence signal detected with anti-ShdA antiserum co-localized with that generated by anti-O4 antiserum in thin sections from the caecum. Expression of the cloned shdA gene from the T7 promoter in vitro resulted in detection of ShdA in the outer membrane of S. Typhimurium and in binding of fibronectin to the bacterial surface. Binding of purified glutathione-S-transferase (GST)-ShdA fusion protein to fibronectin was dose dependent and could be partially inhibited by preincubation with antifibronectin antibodies. GST-ShdA bound to connective tissue and the basement membrane in thin sections from the murine caecum in situ. A similar labelling pattern was produced when thin sections of the murine caecum were stained with antifibronectin antiserum. Collectively, these data demonstrate that ShdA is a surface-localized, fibronectin-binding protein whose expression is induced in vivo in the murine caecum, a tissue in which a cognate receptor of this outer membrane protein is expressed. PMID:11929540

  11. A Multistate Investigation of Antibiotic-Resistant Salmonella enterica Serotype I 4,[5],12:i:- Infections as Part of an International Outbreak Associated with Frozen Feeder Rodents.

    PubMed

    Cartwright, E J; Nguyen, T; Melluso, C; Ayers, T; Lane, C; Hodges, A; Li, X; Quammen, J; Yendell, S J; Adams, J; Mitchell, J; Rickert, R; Klos, R; Williams, I T; Barton Behravesh, C; Wright, J

    2016-02-01

    While most human Salmonella infections result from exposure to contaminated foods, an estimated 11% of all Salmonella infections are attributed to animal exposures, including both direct animal handling and indirect exposures such as cleaning cages and handling contaminated pet food. This report describes the epidemiologic, environmental and laboratory investigations conducted in the United States as part of the response to an international outbreak of tetracycline-resistant Salmonella enterica serotype I 4,[5],12:i:- infections with over 500 illnesses occurring from 2008 to 2010. This investigation found that illness due to the outbreak strain was significantly associated with exposure to pet reptiles and frozen feeder rodents used as food for pet reptiles. Salmonella isolates indistinguishable from the outbreak strain were isolated from a frozen feeder mice-fed reptile owned by a case patient, as well as from frozen feeder mice and environmental samples collected from a rodent producing facility (Company A). An international voluntary recall of all Company A produced frozen feeder animals sold between May 2009 and July 2010 occurred. Only 13% of cases in our investigation were aware of the association between Salmonella infection and mice or rats. Consumers, the pet industry, healthcare providers and veterinarians need to be aware of the potential health risk posed by feeder rodents, whether live or frozen. Frozen feeder rodent producers, suppliers and distributors should follow the animal food labelling requirements as described in 21 CFR §501.5, and all packages of frozen feeder rodents should include safe handling instructions. Persons should wash their hands thoroughly with soap and water after handling live or frozen feeder rodents, as well as reptiles or anything in the area where the animals live. Continued opportunities exist for public health officials, the pet industry, veterinarians and consumers to work together to prevent salmonellosis associated

  12. Outbreak of Salmonella enterica serotype Infantis infection in humans linked to dry dog food in the United States and Canada, 2012.

    PubMed

    Imanishi, Maho; Rotstein, David S; Reimschuessel, Renate; Schwensohn, Colin A; Woody, Dillard H; Davis, Samuel W; Hunt, April D; Arends, Katherine D; Achen, Maya; Cui, Jing; Zhang, Yan; Denny, Lynn F; Phan, Quyen N; Joseph, Lavin A; Tuite, Carla C; Tataryn, Joanne R; Behravesh, Casey Barton

    2014-03-01

    CASE DESCRIPTION--In April 2012, Salmonella enterica serotype Infantis was detected in an unopened bag of dry dog food collected during routine retail surveillance. PulseNet, a national bacterial subtyping network, identified humans with Salmonella Infantis infection with the same genetic fingerprint as the dog food sample. CLINICAL FINDINGS--An outbreak investigation identified 53 ill humans infected with the outbreak strain during January 1 to July 5, 2012, in 21 states and 2 provinces in Canada; 20 (38%) were children ≤ 2 years old, and 12 of 37 (32%) were hospitalized. Of 21 ill people who remembered the dog food brand, 12 (57%) reported a brand produced at a plant in Gaston, SC. Traceback investigations also identified that plant. The outbreak strain was isolated from bags of dry dog food and fecal specimens obtained from dogs that lived with ill people and that ate the implicated dry dog food. TREATMENT AND OUTCOME--The plant was closed temporarily for cleaning and disinfection. Sixteen brands involving > 27,000 metric tons (> 30,000 tons) of dry dog and cat food were recalled. Thirty-one ill dogs linked to recalled products were reported through the FDA consumer complaint system. CLINICAL RELEVANCE-- A one-health collaborative effort on epidemiological, laboratory, and traceback investigations linked dry dog foods produced at a plant to illnesses in dogs and humans. More efforts are needed to increase awareness among pet owners, health-care professionals, and the pet food industry on the risk of illness in pets and their owners associated with dry pet foods and treats. PMID:24548229

  13. The Type VI Secretion System Encoded in Salmonella Pathogenicity Island 19 Is Required for Salmonella enterica Serotype Gallinarum Survival within Infected Macrophages

    PubMed Central

    Blondel, Carlos J.; Jiménez, Juan C.; Leiva, Lorenzo E.; Álvarez, Sergio A.; Pinto, Bernardo I.; Contreras, Francisca; Pezoa, David; Santiviago, Carlos A.

    2013-01-01

    Salmonella enterica serotype Gallinarum is the causative agent of fowl typhoid, a disease characterized by high morbidity and mortality that causes major economic losses in poultry production. We have reported that S. Gallinarum harbors a type VI secretion system (T6SS) encoded in Salmonella pathogenicity island 19 (SPI-19) that is required for efficient colonization of chicks. In the present study, we aimed to characterize the SPI-19 T6SS functionality and to investigate the mechanisms behind the phenotypes previously observed in vivo. Expression analyses revealed that SPI-19 T6SS core components are expressed and produced under in vitro bacterial growth conditions. However, secretion of the structural/secreted components Hcp1, Hcp2, and VgrG to the culture medium could not be determined, suggesting that additional signals are required for T6SS-dependent secretion of these proteins. In vitro bacterial competition assays failed to demonstrate a role for SPI-19 T6SS in interbacterial killing. In contrast, cell culture experiments with murine and avian macrophages (RAW264.7 and HD11, respectively) revealed production of a green fluorescent protein-tagged version of VgrG soon after Salmonella uptake. Furthermore, infection of RAW264.7 and HD11 macrophages with deletion mutants of SPI-19 or strains with genes encoding specific T6SS core components (clpV and vgrG) revealed that SPI-19 T6SS contributes to S. Gallinarum survival within macrophages at 20 h postuptake. SPI-19 T6SS function was not linked to Salmonella-induced cytotoxicity or cell death of infected macrophages, as has been described for other T6SS. Our data indicate that SPI-19 T6SS corresponds to a novel tool used by Salmonella to survive within host cells. PMID:23357385

  14. Proteome of Salmonella Enterica SerotypeTyphimurium Grown in a Low Mg2+/pH Medium

    SciTech Connect

    Shi, Liang; Ansong, Charles; Smallwood, Heather S.; Rommereim, Leah M.; McDermott, Jason E.; Brewer, Heather M.; Norbeck, Angela D.; Taylor, Ronald C.; Gustin, Jean K.; Heffron, Fred; Smith, Richard D.; Adkins, Joshua N.

    2009-09-01

    The facultative intracellular pathogen Salmonella enterica serovar Typhimurium (STM) must replicate within host macrophages in order to establish systemic infection in susceptible mice. In an effort to identify new STM proteins that help the bacterium colonize macrophages, we have cultured STM cells with a low pH/low magnesium medium (MgM) under two different conditions termed MgM-Shock and MgM-Dilution and investigated the impacts of these culturing conditions on the STM proteome by using liquid chromatography-tandem mass spectrometry (LC-MS/MS)-based proteomics. LC-MS/MS results showed that alteration of culturing conditions affected a group of STM proteins differently. Compared to MgM-Shock, MgM-Dilution induced more proteins of the Salmonella-pathogenecity island 2-type III secretion system (SPI2-T3SS). The abundances of the proteins used for cobalamin biosynthesis increased under MgM-Shock condition but decreased under MgM-Dilution condition, while those proteins used for thiamine or biotin biosynthesis were not affected under the former condition but increased under the latter condition. Western-blot (WB) analysis confirmed the LC-MS/MS results. Because cobalamin, thiamine and biotin play different roles in STM metabolism, differential induction of the proteins involved in their biosyntheses suggests that the metabolic states of STM cells under these conditions differ considerably. WB analysis also showed that the abundances of SPI2-T3SS proteins SsaQ and SseE and biotin biosynthesis proteins BioB and BioD increased after STM infection of RAW 264.7 macrophages. Deletion of the gene encoding BioB reduced the ability of STM to replicate inside the macrophages, demonstrating for the first time the involvement of a biotin synthesis protein in STM colonization of macrophages.

  15. Antimicrobial susceptibility and plasmid replicon typing of Salmonella enterica serovar Kentucky isolates recovered from broilers

    Technology Transfer Automated Retrieval System (TEKTRAN)

    In the United States, Salmonella enterica serotype Kentucky has become the predominate serotype recovered from broiler slaughter samples and the prevalence of resistance to streptomycin and tetracycline has increased dramatically in this serotype. To characterize the relationships between antimicro...

  16. Emergence of multidrug-resistant Salmonella enterica serotype Newport infections resistant to expanded-spectrum cephalosporins in the United States.

    PubMed

    Gupta, Amita; Fontana, John; Crowe, Colleen; Bolstorff, Barbara; Stout, Alison; Van Duyne, Susan; Hoekstra, Mike P; Whichard, Jean M; Barrett, Timothy J; Angulo, Frederick J

    2003-12-01

    We describe a field investigation in New England that identified the emergence and epidemiology of new strains of multidrug-resistant Salmonella, Newport-MDRAmpC, and summarize the Center for Disease Control and Prevention's surveillance data for these infections. In Massachusetts, the prevalence of Newport-MDRAmpC among Salmonella serotype Newport isolates obtained from humans increased from 0% (0/14) in 1998 to 53% (32/60) in 2001 (P<.001). In a retrospective case-control study, infection with Newport-MDRAmpC was domestically acquired and was associated with exposure to a dairy farm. Isolates from both humans and cattle had indistinguishable or closely related antibiograms and pulsed-field gel electrophoresis patterns. Nationally, the prevalence of ceftriaxone-resistant Salmonella increased from 0.5% in 1998 to 2.4% in 2001; 85% of the isolates in 2001 were Newport-MDRAmpC, and at least 27 states have isolated these strains from humans, cattle, or ground beef. These data document the widespread emergence of Newport-MDRAmpC strains in the United States and show that the 5-fold increase in the prevalence of Salmonella resistant to expanded-spectrum cephalosporins, between 1998 and 2001, is primarily due to the emergence of Newport-MDRAmpC strains. PMID:14639542

  17. Plane of nutrition influences the performance, innate leukocyte responses, and resistance to an oral Salmonella enterica serotype Typhimurium challenge in Jersey calves.

    PubMed

    Ballou, M A; Hanson, D L; Cobb, C J; Obeidat, B S; Sellers, M D; Pepper-Yowell, A R; Carroll, J A; Earleywine, T J; Lawhon, S D

    2015-03-01

    Two experiments investigated how plane of nutrition influences performance, leukocyte responses, and resistance to an oral Salmonella enterica serotype Typhimurium challenge. In experiment 1, 46 (2±1 d of age) calves were randomly assigned to 2 diets: a low (LPN; n=23) and high plane of nutrition (HPN; n=23). The LPN calves were fed 409 g/d of dry matter (DM) of a 20% crude protein and 20% fat milk replacer, whereas HPN calves were fed 610 and 735 g/d of DM of a 28% crude protein and 25% fat milk replacer during wk 1 and 2 to 6, respectively. In experiment 2, 20 bull calves (LPN; n=11 and HPN; n=9) were orally challenged on d 80 with 1.5×10(7) cfu of Salmonella Typhimurium (ATCC #14028). The HPN calves had a greater incidence (87.5 vs. 45.5%) and duration of days with high fecal scores (5.5 vs. 3.5 d). The LPN calves had greater neutrophil surface expression of L-selectin on d 7, 21, and 42. Following the Salmonella Typhimurium challenge, calf starter DM intake was greater among the HPN calves. The percentage of neutrophils producing an oxidative burst was also greater among HPN calves on d 1 to 5 after the challenge. Similarly, the intensity of the oxidative burst tended to be greater among the HPN calves on d 2 and 3 postchallenge. The secretion of tumor necrosis factor-α from whole-blood cultures stimulated with lipopolysaccharide tended to be greater on d 1 and was greater on d 5 and 6 among HPN calves. The median ranks of haptoglobin concentrations were greater and plasma zinc concentrations tended to be decreased among LPN calves. These data indicate that feeding a HPN to Jersey calves improved average daily gain and feed efficiency, but increased the incidence of high fecal scores during the first few weeks of life; however, the HPN Jersey calves may be more resistant to Salmonella Typhimurium after weaning. PMID:25597967

  18. Metal tolerance in emerging clinically relevant multidrug-resistant Salmonella enterica serotype 4,[5],12:i:- clones circulating in Europe.

    PubMed

    Mourão, Joana; Novais, Carla; Machado, Jorge; Peixe, Luísa; Antunes, Patrícia

    2015-06-01

    The occurrence of acquired metal tolerance genes in emerging MDR Salmonella enterica serotype 4,[5],12:i:- clones was assessed and their associated platforms and tolerance phenotype were characterised. Salmonella 4,[5],12:i:- from different sources belonging to European, Spanish and Southern European clones were studied. Screening for copper (pcoA-pcoD/tcrB), silver/copper (silA-silE), mercury (merA), arsenic (arsB) and tellurite (terF) tolerance genes was performed by PCR/sequencing. CuSO(4)/AgNO(3) MICs were determined in aerobic/anaerobic atmospheres by agar dilution. Conjugation assays, genomic location and plasmid analysis were performed by standard procedures. Most isolates from European (98%) and Spanish (74%) clones carried silA-silE, contrasting with the Southern European clone (26%). merA/62% (European and Spanish clones) and pcoA-pcoD/50% (European clone) were also detected. merA±pco+sil were chromosomally located in the European clone, whereas in Spanish and Southern European clones sil±merA were within plasmids, both with antibiotic resistance genes. The pcoA-pcoD/silA-silE(+) isolates showed higher MICCuSO(4) in anaerobiosis than those without these genes (MIC(50)=24-28 vs. 2 mM). Different MICAgNO(3) of silA-silE(+) (MIC(50)=0.25 mM) and silA-silE(-)(MIC(50)=0.16 mM) isolates were observed in both atmospheres, with an MIC increment after prior exposure to silver (>3 vs. 0.08-0.125 mM) in aerobiosis. A high frequency of copper and silver tolerance, particularly among the two major Salmonella 4,[5],12:i:- MDR clones (European/Spanish) circulating in Europe and causing human infections, might facilitate adaptation/expansion of these strains in metal-contaminated environments, particularly copper in anaerobiosis. Furthermore, metal toxic concentrations in food-animal environments can contribute to persistence of genetic platforms carrying metal/antibiotic resistance genes in this foodborne zoonotic pathogen. PMID:25816978

  19. Interferon-γ and Proliferation Responses to Salmonella enterica Serotype Typhi Proteins in Patients with S. Typhi Bacteremia in Dhaka, Bangladesh

    PubMed Central

    Sheikh, Alaullah; Khanam, Farhana; Sayeed, Md. Abu; Rahman, Taibur; Pacek, Marcin; Hu, Yanhui; Rollins, Andrea; Bhuiyan, Md. Saruar; Rollins, Sean; Kalsy, Anuj; Arifuzzaman, Mohammad; Leung, Daniel T.; Sarracino, David A.; Krastins, Bryan; Charles, Richelle C.; LaRocque, Regina C.; Cravioto, Alejandro; Calderwood, Stephen B.; Brooks, W. Abdullah; Harris, Jason B.; LaBaer, Joshua

    2011-01-01

    Background Salmonella enterica serotype Typhi is a human-restricted intracellular pathogen and the cause of typhoid fever. Cellular immune responses are required to control and clear Salmonella infection. Despite this, there are limited data on cellular immune responses in humans infected with wild type S. Typhi. Methodology/Principal Findings For this work, we used an automated approach to purify a subset of S. Typhi proteins identified in previous antibody-based immuno-affinity screens and antigens known to be expressed in vivo, including StaF-putative fimbrial protein-STY0202, StbB-fimbrial chaperone-STY0372, CsgF-involved in curli production-STY1177, CsgD- putative regulatory protein-STY1179, OppA-periplasmic oligopeptide binding protein precursor-STY1304, PagC-outer membrane invasion protein-STY1878, and conserved hypothetical protein-STY2195; we also generated and analyzed a crude membrane preparation of S. Typhi (MP). In comparison to samples collected from uninfected Bangladeshi and North American participants, we detected significant interferon-γ responses in PBMCs stimulated with MP, StaF, StbB, CsgF, CsgD, OppA, STY2195, and PagC in patients bacteremic with S. Typhi in Bangladesh. The majority of interferon-γ expressing T cells were CD4 cells, although CD8 responses also occurred. We also assessed cellular proliferation responses in bacteremic patients, and confirmed increased responses in infected individuals to MP, StaF, STY2195, and PagC in convalescent compared to acute phase samples and compared to controls. StaF is a fimbrial protein homologous to E. coli YadK, and contains a Pfam motif thought to be involved in cellular adhesion. PagC is expressed in vivo under the control of the virulence-associated PhoP-regulon required for intra-macrophage survival of Salmonella. STY2195 is a conserved hypothetical protein of unknown function. Conclusion/Significance This is the first analysis of cellular immune responses to purified S. Typhi antigens in

  20. Evaluation of a dkgB linked intergenic sequence ribotyping (ISR) method for assigning serotype to Salmonella enterica isolated from poultry environmental samples.

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The Kauffman White (KW) serotyping method requires more than 250 antisera to characterize more than 2,500 Salmonella serovars. The complexity of serotyping could be overcome using molecular methods. In this study, a dkgB-linked intergenic sequence ribotyping (ISR) method that generates sequence occu...

  1. Molecular Characterization of Salmonella enterica Serovars Isolated from a Turkey Production Facility in the Absence of Selective Antimicrobial Pressure.

    PubMed

    Sanad, Yasser M; Johnson, Kelly; Park, Si Hong; Han, Jing; Deck, Joanna; Foley, Steven L; Kenney, Brett; Ricke, Steven; Nayak, Rajesh

    2016-02-01

    This study evaluated antimicrobial resistance and virulence factors in Salmonella enterica isolated from a turkey flock in which the birds were raised in an environment where antimicrobials were not administered to the birds, either through feed or water. Salmonella was isolated from turkeys and various environmental samples in the facility using conventional microbiological procedures. Isolates were serotyped and analyzed phenotypically by antimicrobial resistance profiling and genotypically by pulsed-field gel electrophoresis (PFGE) fingerprinting, integron analysis, plasmid profiling, replicon-based incompatibility (Inc) group typing, and virulence gene profiling. Ninety-five S. enterica isolates were isolated from cecal contents (n = 29), feed (n = 22), leftover feed (n = 13), litter (n = 12), drinkers (n = 10), environment (n = 8), and an insect. The following serotypes were identified: Montevideo (24%), Anatum (22%), Agona (17%), Kentucky and Worthington (12%), Senftenberg (11%), and rough phenotypes (3%). The majority of isolates (61/95; 64%) were susceptible to 12 antimicrobials tested; however, despite the absence of antimicrobials in the facility, approximately 36% of the isolates were resistant to two to five antimicrobials. Class 1 integrons were detected in 8% of the isolates. The integron sequence analysis revealed dihydrofolate reductase (dhfr) and aminoglycoside adenylyl transferase (aadA2) genes, which encode trimethoprim and streptomycin resistance, respectively. Furthermore, 71% of the isolates had at least one plasmid. There were five plasmid replicon types identified among the isolates, including IncI1, IncHI2, IncFIIA, IncB/O, and IncP, with variable prevalence among the serotypes. All 95 isolates tested polymerase chain reaction-positive for 19 virulence genes and negative for virD4 and virB4. The virulence gene profiles were similar within the isolates from the same serotype. Within particular serotypes, PFGE patterns revealed 100

  2. Multiple-Antibiotic Resistance in Salmonella enterica Serotype Paratyphi B Isolates Collected in France between 2000 and 2003 Is Due Mainly to Strains Harboring Salmonella Genomic Islands 1, 1-B, and 1-C

    PubMed Central

    Weill, François-Xavier; Fabre, Laëtitia; Grandry, Bernadette; Grimont, Patrick A. D.; Casin, Isabelle

    2005-01-01

    This study was conducted to investigate the occurrence of multiple-antibiotic resistance among 261 clinical isolates of Salmonella enterica serotype Paratyphi B strains collected between 2000 and 2003 through the network of the French National Reference Center for Salmonella. The 47 multidrug-resistant (MDR) isolates identified (18%), were characterized on the basis of the presence of several resistance genes (blaTEM, blaPSE-1, blaCTX-M, floR, aadA2, qacEΔ1, and sul1), the presence of Salmonella genomic island 1 (SGI1) by PCR mapping and hybridization, and the clonality of these isolates by several molecular (ribotyping, IS200 profiling, and pulsed-field gel electrophoresis [PFGE]) and phage typing methods. The results of PCR and Southern blot experiments indicated that 39 (83%) of the 47 S. enterica serotype Paratyphi B biotype Java MDR isolates possessed the SGI1 cluster (MDR/SGI1). Among these 39 MDR/SGI1 isolates, only 3 contained variations in SGI1, SGI1-B (n = 1) and SGI1-C (n = 2). The 39 MDR/SGI1 isolates showed the same specific PstI-IS200 profile 1, which contained seven copies from 2.6 to 18 kb. Two PstI ribotypes were found in MDR/SGI1 isolates, RP1 (n = 38) and RP6 (n = 1). Ribotype RP1 was also found in two susceptible strains. Analysis by PFGE using XbaI revealed that all the MDR/SGI1 isolates were grouped in two related clusters, with a similarity percentage of 82%. Isolation of MDR/SGI1 isolates in France was observed mainly between the second quarter of 2001 and the end of 2002. The source of the contamination has not been identified to date. A single isolate possessing the extended-spectrum β-lactamase blaCTX-M-15 gene was also identified during the study. PMID:15980351

  3. Salmonella enterica Infections in the United States and Assessment of Coefficients of Variation: A Novel Approach to Identify Epidemiologic Characteristics of Individual Serotypes, 1996–2011

    PubMed Central

    Boore, Amy L.; Hoekstra, R. Michael; Iwamoto, Martha; Fields, Patricia I.; Bishop, Richard D.; Swerdlow, David L.

    2015-01-01

    Background Despite control efforts, salmonellosis continues to cause an estimated 1.2 million infections in the United States (US) annually. We describe the incidence of salmonellosis in the US and introduce a novel approach to examine the epidemiologic similarities and differences of individual serotypes. Methods Cases of salmonellosis in humans reported to the laboratory-based National Salmonella Surveillance System during 1996–2011 from US states were included. Coefficients of variation were used to describe distribution of incidence rates of common Salmonella serotypes by geographic region, age group and sex of patient, and month of sample isolation. Results During 1996–2011, more than 600,000 Salmonella isolates from humans were reported, with an average annual incidence of 13.1 cases/100,000 persons. The annual reported rate of Salmonella infections did not decrease during the study period. The top five most commonly reported serotypes, Typhimurium, Enteritidis, Newport, Heidelberg, and Javiana, accounted for 62% of fully serotyped isolates. Coefficients of variation showed the most geographically concentrated serotypes were often clustered in Gulf Coast states and were also more frequently found to be increasing in incidence. Serotypes clustered in particular months, age groups, and sex were also identified and described. Conclusions Although overall incidence rates of Salmonella did not change over time, trends and epidemiological factors differed remarkably by serotype. A better understanding of Salmonella, facilitated by this comprehensive description of overall trends and unique characteristics of individual serotypes, will assist in responding to this disease and in planning and implementing prevention activities. PMID:26701276

  4. Serotype and Antimicrobial Resistance Patterns of Salmonella Isolates from Commercial Birds and Poultry Environment in Mississippi

    Technology Transfer Automated Retrieval System (TEKTRAN)

    To obtain information about Salmonella from commercial birds and poultry environments within Mississippi, 50 Salmonella enterica isolates were collected and characterized by Intergenic Sequence Ribotyping (ISR) serotyping and by determining antimicrobial resistance. ISR assigned serotype to all 50 S...

  5. Genomic investigation of Salmonella enterica sequences associated with long-term colonization of the bovine gut

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Salmonella enterica subsp. enterica is a leading cause of food and waterborne infections globally in both humans and livestock with an estimated 93 million annual human infections caused by nontyphoidal S. enterica alone. However, some serotypes within this species are known to cause mild infection...

  6. Only one of the two type VI secretion systems encoded in the Salmonella enterica serotype Dublin genome is involved in colonization of the avian and murine hosts.

    PubMed

    Pezoa, David; Blondel, Carlos J; Silva, Cecilia A; Yang, Hee-Jeong; Andrews-Polymenis, Helene; Santiviago, Carlos A; Contreras, Inés

    2014-01-01

    The type VI secretion system (T6SS) is a virulence factor for many Gram-negative bacteria. Salmonella genus harbors five phylogenetically distinct T6SS loci encoded in Salmonella Pathogenicity Islands (SPIs) SPI-6, SPI-19, SPI-20, SPI-21 and SPI-22, which are differentially distributed among serotypes. The T6SSs encoded in SPI-6 and SPI-19 contribute to pathogenesis of serotypes Typhimurium and Gallinarum in mice and chickens, respectively. Salmonella Dublin is a pathogen restricted to cattle where it causes a systemic disease. Also, it can colonize other hosts such as chickens and mice, which can act as reservoirs of this serotype. Salmonella Dublin harbors the genes for both T6SS(SPI-6) and T6SS(SPI-19). This study has determined the contribution of T6SS(SPI-6) and T6SS(SPI-19) to host-colonization by Salmonella Dublin using avian and murine models of infection. Competitive index experiments showed that, a mutant strain lacking both T6SSs (∆T6SS(SPI-6)/∆T6SS(SPI-19)) presents a strong colonization defect in cecum of chickens, similar to the defect observed for the ∆T6SS(SPI-6) mutant, suggesting that this serotype requires a functional T6SS(SPI-6) for efficient colonization of the avian gastrointestinal tract. Colonization of mice was also defective, although to a lesser extent than in chickens. In contrast, the T6SS(SPI-19) was not necessary for colonization of either chickens or mice. Transfer of T6SS(SPI-6), but not T6SS(SPI-19), restored the ability of the double mutant to colonize both animal hosts. Our data indicate that Salmonella Dublin requires only the T6SS(SPI-6) for efficient colonization of mice and chickens, and that the T6SS(SPI-6) and T6SS(SPI-19) are not functionally redundant. PMID:24405577

  7. The Genomic Blueprint of Salmonella enterica subspecies enterica serovar Typhi P-stx-12

    PubMed Central

    Ong, Su Yean; Pratap, Chandra Bhan; Wan, Xuehua; Hou, Shaobin; Rahman, Ahmad Yamin Abdul; Saito, Jennifer A.; Nath, Gopal; Alam, Maqsudul

    2013-01-01

    Salmonella enterica subspecies enterica serovar Typhi is a rod-shaped, Gram-negative, facultatively anaerobic bacterium. It belongs to the family Enterobacteriaceae in the class Gammaproteobacteria, and has the capability of residing in the human gallbladder by forming a biofilm and hence causing the person to become a typhoid carrier. Here we present the complete genome of Salmonella enterica subspecies enterica serotype Typhi strain P-stx-12, which was isolated from a chronic carrier in Varanasi, India. The complete genome comprises a 4,768,352 bp chromosome with a total of 98 RNA genes, 4,691 protein-coding genes and a 181,431 bp plasmid. Genome analysis revealed that the organism is closely related to Salmonella enterica serovar Typhi strain Ty2 and Salmonella enterica serovar Typhi strain CT18, although their genome structure is slightly different. PMID:24019994

  8. Multilocus variable-number of tandem-repeats analysis of Salmonella enterica serotype Gallinarum and comparison with pulsed-field gel electrophoresis genotyping.

    PubMed

    Bergamini, Federica; Iori, Alessandra; Massi, Paola; Pongolini, Stefano

    2011-05-01

    Salmonella enterica serovar Gallinarum is the causative agent of fowl typhoid, a severe disease of poultry, responsible for heavy economic losses. Epidemiologic investigation of fowl typhoid significantly benefits from molecular typing tools, RAPD and PFGE have been proposed for this purpose. PFGE, a well established technique, is still the gold standard among typing methods for most bacteria, including salmonella. Nevertheless, it has some limitations regarding execution and reproducibility, in particular it is labour intensive and requires good technical expertise. Furthermore, it needs accurate standardization and results can be ambiguous to interpret. Such limitations can hamper reproducibility and transfer of results. As a possible alternative to PFGE, multilocus variable-number of tandem-repeats analysis (MLVA) has recently emerged as an effective genotyping method for many bacterial pathogens showing high discriminatory power associated to robustness. We developed a six-loci MLVA protocol for Salmonella Gallinarum and compared it to PFGE performed with SpeI, XbaI and NotI on fifty isolates. The proposed MLVA has a high discriminatory power, equivalent to that of the three-enzyme PFGE (Simpson's index 0.94 for MLVA, 0.93 for three-enzyme PFGE) but it is simpler to perform and straightforward in genotype identification, allowing unambiguous exchange of results. Stability of selected VNTR loci, assessed in vitro and in vivo, is good but not absolute, reflecting the sensitivity of MLVA to detect evolutionary changes of bacteria. Clustering of the isolates as determined by MLVA typing is substantially confirmed by PFGE. PMID:21208755

  9. An outbreak due to peanuts in their shell caused by Salmonella enterica serotypes Stanley and Newport--sharing molecular information to solve international outbreaks.

    PubMed Central

    Kirk, M. D.; Little, C. L.; Lem, M.; Fyfe, M.; Genobile, D.; Tan, A.; Threlfall, J.; Paccagnella, A.; Lightfoot, D.; Lyi, H.; McIntyre, L.; Ward, L.; Brown, D. J.; Surnam, S.; Fisher, I. S. T.

    2004-01-01

    Salmonellosis is a global problem caused by the international movement of foods and high incidence in exporting countries. In September 2001, in an outbreak investigation Australia isolated Salmonella Stanley from imported peanuts, which resulted in a wider investigation in Canada, England & Wales and Scotland. Patients infected with Salmonella serotypes known to be isolated from peanuts and reported to surveillance systems were interviewed to determine exposure histories. Tagged image file format (TIFF) images of pulsed-field gel electrophoresis (PFGE) patterns of Salmonella isolates were shared electronically amongst laboratories. Laboratories tested packets of 'Brand X' peanuts from various lots and product lines. In total, 97 cases of S. Stanley and 12 cases of S. Newport infection were found. Seventy-three per cent (71/97) of S. Stanley cases were in persons of Asian ethnicity. Twenty-eight per cent of cases recalled eating Brand X peanuts and a further 13% had peanuts in their house in the previous month or had eaten Asian-style peanuts. Laboratories isolated S. Stanley, S. Newport, S. Kottbus, S. Lexington and S. Unnamed from Brand X peanuts. Isolates of S. Stanley from peanuts and human patients were indistinguishable by PFGE. This international outbreak resulted from a product originating from one country affecting several others. Rapid sharing of electronic DNA images was a crucial factor in delineating the outbreak; multinational investigations would benefit from a harmonized approach. PMID:15310157

  10. Molecular characterization of Salmonella enterica serotype Enteritidis isolates from humans by antimicrobial resistance, virulence genes, and pulsed-field gel electrophoresis.

    PubMed

    Zou, Ming; Keelara, Shivaramu; Thakur, Siddhartha

    2012-03-01

    Salmonella enterica serovar Enteritidis (S. Enteritidis) is a major serovar associated with human salmonellosis. A total of 425 clinical S. Enteritidis isolates of human origin were collected between June 2009 and September 2010 from North Carolina. The isolates were further characterized for antimicrobial susceptibility, antimicrobial resistance coding determinants, virulence genes, and fingerprint profiles to determine whether they were similar or different to the S. Enteritidis strain responsible for the human outbreak due to consumption of contaminated eggs. Ten different antimicrobial resistance phenotypes were observed with the highest frequency of resistance exhibited to ampicillin (n=10; 2.35%). The isolates were predominantly pansusceptible (n=409; 96.23%); however, seven isolates were multidrug resistant (MDR; i.e., resistant to three or more antimicrobials). Extended spectrum β-lactamase (ESBL) coding genes (bla(TEM) and bla(PSE)) were detected in the ampicillin-resistant isolates, whereas a single MDR isolate tested positive for class 1 integron (1 kb). The majority of the isolates (n=422; 99.3%) carried the invA, mgtC, stn, sopB, sopE1, and sefA virulence genes. However, 37 (8.7%) and 46 (10.82%) S. Enteritidis isolates tested negative for the plasmid encoded genes spvC and rck, respectively. Pulsed-field gel electrophoresis (PFGE) typing of 118 S. Enteritidis isolates by restriction enzymes XbaI and BlnI resulted in seven clusters, each with a discriminatory index (DI) of 0.715 and 0.785, respectively. The combination of XbaI-BlnI patterns generated a dendrogram with 14 clusters and a higher DI of 0.914. The PFGE profile of 80 isolates matched 100% with the S. Enteritidis strain that has been cited for the recent outbreak in the United States due to consumption of contaminated eggs. In conclusion, we identified a genotypic similar S. Enteritidis population in our study based on antimicrobial susceptibility, virulence gene, and PFGE fingerprint

  11. Increasing Ceftriaxone Resistance in Salmonellae, Taiwan

    PubMed Central

    Su, Lin-Hui; Teng, Wen-Shin; Chen, Chyi-Liang; Lee, Hao-Yuan; Li, Hsin-Chieh; Wu, Tsu-Lan

    2011-01-01

    In Taiwan, despite a substantial decline of Salmonella enterica serotype Choleraesuis infections, strains resistant to ciprofloxacin and ceftriaxone persist. A self-transferable blaCMY-2-harboring IncI1 plasmid was identified in S. enterica serotypes Choleraesuis, Typhimurium, Agona, and Enteritidis and contributed to the overall increase of ceftriaxone resistance in salmonellae. PMID:21749777

  12. transcriptional response of pigs to Salmonella infection: Comparison of responses to infection with Salmonella eimerica serotype Typhimurium as that observed in S. choleraesuis infection.

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Swine responses to, and control of, Salmonella enterica serotype Typhimurium (ST) infection have been compared to S. enterica serotype Choleraesuis (SC) infection. Using subtractive suppression hybridization (SSH), long oligonucleotide Qiagen and Affymetrix porcine GeneChip® arrays, and real time ge...

  13. Draft Genome Sequences of 33 Salmonella enterica Clinical and Wildlife Isolates from Chile

    PubMed Central

    Toro, Magaly; Allard, Marc; Brown, Eric W.; Evans, Peter

    2015-01-01

    Salmonella enterica causes health problem worldwide. The relationships among strains that are from the same serotype but different hosts, countries, and continents remain elusive. Few genome sequences are available from S. enterica isolates from South America. Therefore, we sequenced the genomes of 33 strains from diverse sources isolated in Chile and determined that they were of different serotypes. These genomes will improve phylogenetic analysis of Salmonella strains from Chile and the rest of South America. PMID:25792040

  14. Genome sequences of ten Salmonella enterica serovars isolated from a single dairy farm

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Here we report draft genomes of twenty-seven isolates of Salmonella enterica subsp. enterica representing the seven serotypes isolated from cows in a Pennsylvania dairy herd, the farm on which they were reared, and the associated off-site heifer-raising facility over an eight year sampling period. ...

  15. Salmonella enterica.

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Avian Salmonella infections are important as both a cause of clinical disease in poultry and as a source of food-borne transmission of disease to humans. Host-adapted salmonellae (Salmonella enterica serovar Pullorum and Gallinarum) are responsible for severe systemic diseases, whereas numerous sero...

  16. Subtyping of Salmonella enterica subspecies I using single nucleotide polymorphisms in adenylate cyclase (cyaA)

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Methods to rapidly identify serotypes of Salmonella enterica subspecies I are of vital importance for protecting the safety of food. To supplement the serotyping method dkgB-linked intergenic sequence ribotyping (ISR), single nucleotide polymorphisms (SNPs) were characterized within adenylate cyclas...

  17. Draft Whole-Genome Sequences of 25 Salmonella enterica Strains Representing 24 Serovars

    PubMed Central

    Brumwell, Stephanie L.; Lingohr, Erika J.; Ahmad, Aaminah; Blimkie, Travis M.; Kogan, Benjamin A.; Pilsworth, Jessica; Rehman, Muhammad A.; Schleicher, Krista L.; Shanmugaraj, Jenitta; Kropinski, Andrew M.; Nash, John H. E.

    2016-01-01

    We report the draft genome sequences of 25 Salmonella enterica strains representing 24 different serotypes, many of which were not available in public repositories during our selection process. These draft genomes will provide useful reference for the genetic variation between serotypes and aid in the development of molecular typing tools. PMID:26941156

  18. Nationwide pseudo-outbreak of Salmonella enterica ssp. diarizonae, France.

    PubMed

    Thiolet, J M; Jourdan-Da Silva, N; Reggiani, A; De Valk, H; Coignard, B; Weill, F X

    2011-06-01

    To investigate an increased incidence of human cultures growing Salmonella enterica ssp. diarizonae serotype 61:k:1,5,7 in France in 2008, we reviewed medical records of case patients and identified the material used during invasive procedures and for bacterial culture. Trace-back investigations incriminated culture media containing contaminated sheep blood agar. PMID:20718799

  19. Foodborne Outbreak and Nonmotile Salmonella enterica Variant, France

    PubMed Central

    Brisabois, Anne; Accou-Demartin, Marie; Josse, Adeline; Marault, Muriel; Francart, Sylvie; Da Silva, Nathalie Jourdan; Weill, François-Xavier

    2012-01-01

    We report a food-related outbreak of salmonellosis in humans caused by a nonmotile variant of Salmonella enterica serotype Typhimurium in France in 2009. This nonmotile variant had been circulating in laying hens but was not considered as Typhimurium and consequently escaped European poultry flock regulations. PMID:22257550

  20. Salmonella enterica genomics and genetics of antimicrobial resistance

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Salmonella enterica is an important food-borne pathogen and an excellent model system for the study of genomics, virulence and pathogenesis. . There are over 2,400 Salmonella serotypes each of which differs in their ability to cause disease in humans and animals, persist within the host, and survive...

  1. Effects of integrated treatment of nonthermal UV-C light and different antimicrobial wash on Salmonella enterica on plum tomatoes

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Produce contamination by foodborne pathogens remains a serious threat. This study investigated synergistic effects of ultraviolet-C and various active sanitizers’ washes against Salmonella enterica on plum tomatoes. A bacterial cocktail containing three serotypes of Salmonella enterica (S. Newport H...

  2. High-throughput Molecular Determination of Salmonella enterica Serovars Use of Multiplex PCR and Capillary Electrophoresis Analysis

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Salmonella enterica is a zoonotic pathogen and is a leading cause of food-borne illness worldwide. There are over 2,500 serotypes of Salmonella reported. Identification of the serotype is key in defining the etiological agent during an outbreak investigation. In the current study, a high-throughput ...

  3. Development of Microarray and Multiplex Polymerase Chain Reaction Assays for Identification of Serovars and Virulence Genes in Salmonella enterica of Human or Animal Origin

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Salmonella enterica is an important enteric pathogen consisting of many serotypes that can cause severe clinical diseases in animals and humans. Rapid identification of Salmonella isolates is especially important for epidemiological monitoring and controlling outbreaks of disease. Although immunolo...

  4. Prevalence and Characteristics of Salmonella Serotypes Isolated from Fresh Produce Marketed in the United States.

    PubMed

    Reddy, Shanker P; Wang, Hua; Adams, Jennifer K; Feng, Peter C H

    2016-01-01

    Salmonella continues to rank as one of the most costly foodborne pathogens, and more illnesses are now associated with the consumption of fresh produce. The U.S. Department of Agriculture Microbiological Data Program (MDP) sampled select commodities of fresh fruit and vegetables and tested them for Salmonella, pathogenic Escherichia coli, and Listeria. The Salmonella strains isolated were further characterized by serotype, antimicrobial resistance, and pulsed-field gel electrophoresis profile. This article summarizes the Salmonella data collected by the MDP between 2002 and 2012. The results show that the rates of Salmonella prevalence ranged from absent to 0.34% in cilantro. A total of 152 isolates consisting of over 50 different serotypes were isolated from the various produce types, and the top five were Salmonella enterica serotype Cubana, S. enterica subspecies arizonae (subsp. IIIa) and diarizonae (subsp. IIIb), and S. enterica serotypes Newport, Javiana, and Infantis. Among these, Salmonella serotypes Newport and Javiana are also listed among the top five Salmonella serotypes that caused most foodborne outbreaks. Other serotypes that are frequent causes of infection, such as S. enterica serotypes Typhimurium and Enteritidis, were also found in fresh produce but were not prevalent. About 25% of the MDP samples were imported produce, including 65% of green onions, 44% of tomatoes, 42% of hot peppers, and 41% of cantaloupes. However, imported produce did not show higher numbers of Salmonella-positive samples, and in some products, like cilantro, all of the Salmonella isolates were from domestic samples. About 6.5% of the Salmonella isolates were resistant to the antimicrobial compounds tested, but no single commodity or serotype was found to be the most common carrier of resistant strains or of resistance. The pulsed-field gel electrophoresis profiles of the produce isolates showed similarities with Salmonella isolates from meat samples and from outbreaks, but

  5. Increased water activity reduces the thermal resistance of Salmonella enterica in peanut butter.

    PubMed

    He, Yingshu; Li, Ye; Salazar, Joelle K; Yang, Jingyun; Tortorello, Mary Lou; Zhang, Wei

    2013-08-01

    Increased water activity in peanut butter significantly (P < 0.05) reduced the heat resistance of desiccation-stressed Salmonella enterica serotypes treated at 90 °C. The difference in thermal resistance was less notable when strains were treated at 126 °C. Using scanning electron microscopy, we observed minor morphological changes of S. enterica cells resulting from desiccation and rehydration processes in peanut oil. PMID:23728806

  6. Rapid detection of Salmonella enterica with primers specific for iroB.

    PubMed Central

    Bäumler, A J; Heffron, F; Reissbrodt, R

    1997-01-01

    The iroB gene of Salmonella enterica is absent from the chromosome of the related organism Escherichia coli. We determined the distribution of this gene among 150 bacterial isolates, representing 51 serotypes of different Salmonella species and subspecies and 8 other bacterial species which are frequent contaminants during routine enrichment procedures by Southern hybridization. An iroB-specific DNA probe detected homologous sequences in all strains of S. enterica, including serotypes of S. enterica subsp. enterica (I), salamae (II), diarizonae (IIIb), and houtenae (IV). No hybridization signal was obtained with strains of Salmonella bongori or other bacterial species. In contrast, hybridization with a DNA probe specific for purD, a purine biosynthesis gene, detected homologs in all bacterial species tested. Primers specific for iroB were used to amplify this gene from 197 bacterial isolates by PCR. The iroB gene could be PCR amplified from S. enterica subsp. enterica (I), salamae (II), diarizonae (IIIb), houtenae (IV), arizonae (IIIa), and indica (VI), but not from S. bongori or other bacterial species. Thus, PCR amplification of iroB can be used to distinguish between S. enterica and other bacterial species, including S. bongori. A combination of preenrichment in buffered peptone water supplemented with ferrioxamine E and amplification of iroB by magnetic immuno-PCR allowed detection of S. enterica in albumen within 24 h. In conclusion, PCR amplification of iroB is a new sensitive and selective method which has the potential to rapidly detect S. enterica serotypes. PMID:9114411

  7. Genome wide evolutionary analyses reveal serotype specific patterns of positive selection in selected Salmonella serotypes

    PubMed Central

    2009-01-01

    Background The bacterium Salmonella enterica includes a diversity of serotypes that cause disease in humans and different animal species. Some Salmonella serotypes show a broad host range, some are host restricted and exclusively associated with one particular host, and some are associated with one particular host species, but able to cause disease in other host species and are thus considered "host adapted". Five Salmonella genome sequences, representing a broad host range serotype (Typhimurium), two host restricted serotypes (Typhi [two genomes] and Paratyphi) and one host adapted serotype (Choleraesuis) were used to identify core genome genes that show evidence for recombination and positive selection. Results Overall, 3323 orthologous genes were identified in all 5 Salmonella genomes analyzed. Use of four different methods to assess homologous recombination identified 270 genes that showed evidence for recombination with at least one of these methods (false discovery rate [FDR] <10%). After exclusion of genes with evidence for recombination, site and branch specific models identified 41 genes as showing evidence for positive selection (FDR <20%), including a number of genes with confirmed or likely roles in virulence and ompC, a gene encoding an outer membrane protein, which has also been found to be under positive selection in other bacteria. A total of 8, 16, 7, and 5 genes showed evidence for positive selection in Choleraesuis, Typhi, Typhimurium, and Paratyphi branch analyses, respectively. Sequencing and evolutionary analyses of four genes in an additional 42 isolates representing 23 serotypes confirmed branch specific positive selection and recombination patterns. Conclusion Our data show that, among the four serotypes analyzed, (i) less than 10% of Salmonella genes in the core genome show evidence for homologous recombination, (ii) a number of Salmonella genes are under positive selection, including genes that appear to contribute to virulence, and (iii

  8. Genome-Scale Screening and Validation of Targets for Identification of Salmonella enterica and Serovar Prediction.

    PubMed

    Zhou, Xiujuan; Zhang, Lida; Shi, Chunlei; Fratamico, Pina M; Liu, Bin; Paoli, George C; Dan, Xianlong; Zhuang, Xiaofei; Cui, Yan; Wang, Dapeng; Shi, Xianming

    2016-03-01

    Salmonella enterica is the most common foodborne pathogen worldwide, with 2,500 recognized serovars. Detection of S. enterica and its classification into serovars are essential for food safety surveillance and clinical diagnosis. The PCR method is useful for these applications because of its rapidity and high accuracy. We obtained 412 candidate detection targets for S. enterica using a comparative genomics mining approach. Gene ontology (GO) functional enrichment analysis of these candidate targets revealed that the GO term with the largest number of unigenes with known function (38 of 177, 21.5%) was significantly involved in pathogenesis (P < 10(-24)). All the candidate targets were then evaluated by PCR assays. Fifteen targets showed high specificity for the detection of S. enterica by verification with 151 S. enterica strains and 34 non-Salmonella strains. The phylogenetic trees of verified targets were highly comparable with those of housekeeping genes, especially for differentiating S. enterica strains into serovars. The serovar prediction ability was validated by sequencing one target (S9) for 39 S. enterica strains belonging to six serovars. Identical mutation sites existed in the same serovar, and different mutation sites were found in diverse serovars. Our findings revealed that 15 verified targets can be potentially used for molecular detection, and some of them can be used for serotyping of S. enterica strains. PMID:26939647

  9. Draft Genome Sequences of 37 Salmonella enterica Strains Isolated from Poultry Sources in Nigeria.

    PubMed

    Useh, Nicodemus M; Ngbede, Emmanuel O; Akange, Nguavese; Thomas, Milton; Foley, Andrew; Keena, Mitchel Chan; Nelson, Eric; Christopher-Hennings, Jane; Tomita, Masaru; Suzuki, Haruo; Scaria, Joy

    2016-01-01

    Here, we report the availability of draft genomes of several Salmonella serotypes, isolated from poultry sources from Nigeria. These genomes will help to further understand the biological diversity of S. enterica and will serve as references in microbial trace-back studies to improve food safety. PMID:27151793

  10. Draft Genome Sequences of 37 Salmonella enterica Strains Isolated from Poultry Sources in Nigeria

    PubMed Central

    Useh, Nicodemus M.; Ngbede, Emmanuel O.; Akange, Nguavese; Thomas, Milton; Foley, Andrew; Keena, Mitchel Chan; Nelson, Eric; Christopher-Hennings, Jane; Tomita, Masaru

    2016-01-01

    Here, we report the availability of draft genomes of several Salmonella serotypes, isolated from poultry sources from Nigeria. These genomes will help to further understand the biological diversity of S. enterica and will serve as references in microbial trace-back studies to improve food safety. PMID:27151793

  11. Molecular characterization of Salmonella enterica isolates associated with starling-livestock interactions.

    PubMed

    Carlson, James C; Hyatt, Doreene R; Bentler, Kevin; Mangan, Anna M; Russell, Michael; Piaggio, Antoinette J; Linz, George M

    2015-08-31

    Bird-livestock interactions have been implicated as potential sources for bacteria within concentrated animal feeding operations (CAFO). In this study we characterized XbaI-digested genomic DNA from Salmonella enterica using pulsed-field gel electrophoresis (PFGE). The PFGE analysis was conducted using 182 S. enterica isolates collected from a single CAFO between 2009 and 2012. Samples collected in 2012 were subjected to antimicrobial susceptibility testing. The analysis was limited to S. enterica serotypes, with at least 10 isolates, known to occur in both European starlings (Sturnus vulgaris) and cattle (Bos taurus) within this CAFO. A total of five different serotypes were screened; S. Anatum, S. Kentucky, S. Meleagridis, S. Montevideo, S. Muenchen. These samples were recovered from five different sample types; starling gastrointestinal tracts (GI), starling external wash, cattle feces, cattle feed and cattle water troughs. Indistinguishable S. enterica PFGE profiles were recovered from isolates originating in all sample types. Antimicrobial resistance (AMR) was also associated with indistinguishable S. enterica isolates recovered from all samples types. These data suggests that AMR S. enterica is transmitted between cattle and starlings and that shared feed sources are likely contributing to infections within both species. Moreover we isolated indistinguishable PFGE profiles across all years of data collection, suggesting long-term environmental persistence may be mediated by starling visits to CAFO. PMID:25866128

  12. Subtyping of Salmonella enterica Subspecies I Using Single-Nucleotide Polymorphisms in Adenylate Cyclase.

    PubMed

    Guard, Jean; Abdo, Zaid; Byers, Sara Overstreet; Kriebel, Patrick; Rothrock, Michael J

    2016-07-01

    Methods to rapidly identify serotypes of Salmonella enterica subspecies I are of vital importance for protecting the safety of food. To supplement the serotyping method dkgB-linked intergenic sequence ribotyping (ISR), single-nucleotide polymorphisms were characterized within adenylate cyclase (cyaA). The National Center for Biotechnology Information (NCBI) database had 378 cyaA sequences from S. enterica subspecies I, which included 42 unique DNA sequences and 19 different amino acid sequences. Five representative isolates, namely serotypes Typhimurium, Kentucky, Enteritidis phage type PT4, and two variants of Enteritidis phage type PT13a, were differentiated within a microsphere-based fluidics system in cyaA by allele-specific primer extension. Validation against 25 poultry-related environmental Salmonella isolates representing 11 serotypes yielded a ∼89% success rate at identifying the serotype of the isolate, and a different region could be targeted to achieve 100%. When coupled with ISR, all serotypes were differentiated. Phage lineages of serotype Enteritidis 13a and 4 were identified, and a biofilm-forming strain of PT13a was differentiated from a smooth phenotype within phage type. Comparative ranking of mutation indices to genes such as the tRNA transferases, the diguanylate cyclases, and genes used for multilocus sequence typing indicated that cyaA is an appropriate gene for assessing epidemiological trends of Salmonella because of its relative stability in nucleotide composition. PMID:27035032

  13. Subtyping of Salmonella enterica Subspecies I Using Single-Nucleotide Polymorphisms in Adenylate Cyclase

    PubMed Central

    Abdo, Zaid; Byers, Sara Overstreet; Kriebel, Patrick; Rothrock, Michael J.

    2016-01-01

    Abstract Methods to rapidly identify serotypes of Salmonella enterica subspecies I are of vital importance for protecting the safety of food. To supplement the serotyping method dkgB-linked intergenic sequence ribotyping (ISR), single-nucleotide polymorphisms were characterized within adenylate cyclase (cyaA). The National Center for Biotechnology Information (NCBI) database had 378 cyaA sequences from S. enterica subspecies I, which included 42 unique DNA sequences and 19 different amino acid sequences. Five representative isolates, namely serotypes Typhimurium, Kentucky, Enteritidis phage type PT4, and two variants of Enteritidis phage type PT13a, were differentiated within a microsphere-based fluidics system in cyaA by allele-specific primer extension. Validation against 25 poultry-related environmental Salmonella isolates representing 11 serotypes yielded a ∼89% success rate at identifying the serotype of the isolate, and a different region could be targeted to achieve 100%. When coupled with ISR, all serotypes were differentiated. Phage lineages of serotype Enteritidis 13a and 4 were identified, and a biofilm-forming strain of PT13a was differentiated from a smooth phenotype within phage type. Comparative ranking of mutation indices to genes such as the tRNA transferases, the diguanylate cyclases, and genes used for multilocus sequence typing indicated that cyaA is an appropriate gene for assessing epidemiological trends of Salmonella because of its relative stability in nucleotide composition. PMID:27035032

  14. Molecular fingerprinting of Salmonella enterica subsp. enterica Typhimurium and Salmonella enterica subsp. enterica derby isolated from tropical seafood in South India.

    PubMed

    Kumar, Rakesh; Surendran, P K; Thampuran, Nirmala

    2008-09-01

    Salmonella enterica subsp. enterica Typhimurium and Salmonella enterica subsp. enterica Derby strains isolated from different seafood were genotyped by PCR-ribotyping and ERIC-PCR assays. This study has ascertained the genetic relatedness among serovars prevalent in tropical seafood. PCR-ribotyping exhibited genetic variation in both Salmonella serovars, and ribotype profile (II) was most predominant, which was observed in 10/18 of Salmonella enterica subsp. enterica Typhimurium and 7/17 Salmonella enterica subsp. enterica Derby isolates. Cluster analysis of ERIC-PCR for Salmonella enterica subsp. enterica Typhimurium strains exhibited nine different banding patterns and four strains showed >95% genetic homology within the cluster pairs. ERIC-PCR produced more genetic variations in Salmonella enterica subsp. enterica Typhimurium; nevertheless, both methods were found to be comparable for Salmonella enterica subsp. enterica Derby isolates. Discrimination index of PCR-ribotyping for Salmonella enterica subsp. enterica Typhimurium isolates was obtained at 0.674 and index value 0.714 was observed for Salmonella enterica subsp. enterica Derby strains. Molecular fingerprinting investigation highlighted the hypothesis of diverse routes of Salmonella contamination in seafood as multiple clones of Salmonella enterica subsp. enterica Typhimurium and Salmonella enterica subsp. enterica Derby were detected in same or different seafood throughout the study period. PMID:18480975

  15. Mechanisms of antimicrobial resistant Salmonella enterica transmission associated with starling-livestock interactions.

    PubMed

    Carlson, James C; Hyatt, Doreene R; Ellis, Jeremy W; Pipkin, David R; Mangan, Anna M; Russell, Michael; Bolte, Denise S; Engeman, Richard M; DeLiberto, Thomas J; Linz, George M

    2015-08-31

    Bird-livestock interactions have been implicated as potential sources for bacteria within concentrated animal feeding operations (CAFO). European starlings (Sturnus vulgaris) in particular are known to contaminate cattle feed and water with Salmonella enterica through their fecal waste. We propose that fecal waste is not the only mechanisms through which starlings introduce S. enterica to CAFO. The goal of this study was to assess if starlings can mechanically move S. enterica. We define mechanical movement as the transportation of media containing S. enterica, on the exterior of starlings within CAFO. We collected 100 starlings and obtained external wash and gastrointestinal tract (GI) samples. We also collected 100 samples from animal pens. Within each pen we collected one cattle fecal, feed, and water trough sample. Isolates from all S. enterica positive samples were subjected to antimicrobial susceptibility testing. All sample types, including 17% of external starling wash samples, contained S. enterica. All sample types had at least one antimicrobial resistant (AMR) isolate and starling GI samples harbored multidrug resistant S. enterica. The serotypes isolated from the starling external wash samples were all found in the farm environment and 11.8% (2/17) of isolates from positive starling external wash samples were resistant to at least one class of antibiotics. This study provides evidence of a potential mechanism of wildlife introduced microbial contamination in CAFO. Mechanical movement of microbiological hazards, by starlings, should be considered a potential source of bacteria that is of concern to veterinary, environmental and public health. PMID:25960334

  16. Synthetic brominated furanone F202 prevents biofilm formation by potentially human pathogenic Escherichia coli O103:H2 and Salmonella ser. Agona on abiotic surfaces

    PubMed Central

    Vestby, LK; Johannesen, KCS; Witsø, IL; Habimana, O; Scheie, AA; Urdahl, AM; Benneche, T; Langsrud, S; Nesse, LL

    2014-01-01

    Aims Investigate the use of a synthetic brominated furanone (F202) against the establishment of biofilm by Salmonella ser. Agona and E. coli O103:H2 under temperature conditions relevant for the food and feed industry as well as under temperature conditions optimum for growth. Methods and Results Effect of F202 on biofilm formation by Salmonella ser. Agona and E. coli O103:H2 was evaluated using a microtiter plate assay and confocal microscopy. Effect of F202 on bacterial motility was investigated using swimming and swarming assays. Influence on flagellar synthesis by F202 was examined by flagellar staining. Results showed that F202 inhibited biofilm formation without being bactericidal. F202 was found to affect both swimming and swarming motility without, however, affecting the expression of flagella. Conclusions F202 showed its potential as a biofilm inhibitor of Salmonella ser. Agona and E. coli O103:H2 under temperature conditions relevant for the feed and food industry as well as temperatures optimum for growth. One potential mode of action of F202 was found to be by targeting flagellar function. Significance and Impact of the Study The present study gives valuable new knowledge to the potential use of furanones as a tool in biofilm management in the food and feed industry. PMID:24118802

  17. Identification of multidrug-resistant Salmonella enterica serovar typhimurium isolates that have an antibiotic-induced invasion phenotype

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Multidrug-resistant (MDR) Salmonella is an important food safety issue in humans and animals. The National Antimicrobial Resistance Monitoring System (NARMS) has reported that 27.3% of Salmonella enterica serotype Typhimurium isolates in humans were resistant to three or more classes of antibiotics...

  18. First Report of Human Infection with Salmonella enterica Serovar Apapa Resulting from Exposure to a Pet Lizard▿

    PubMed Central

    Cooke, Fiona J.; De Pinna, Elizabeth; Maguire, Clare; Guha, Simantee; Pickard, Derek J.; Farrington, Mark; Threlfall, E. John

    2009-01-01

    We present the first documented human case of Salmonella enterica serovar Apapa infection, isolated concurrently from a hospital inpatient and a pet lizard. The isolates were identical by biochemical profiling and pulsed-field gel electrophoresis. This rare serotype is known to be associated with reptiles. The current practice for avoiding reptile-associated infections is reviewed. PMID:19535527

  19. Antimicrobial Resistance of Salmonella enterica Isolates from Tonsil and Jejunum with Lymph Node Tissues of Slaughtered Swine in Metro Manila, Philippines

    PubMed Central

    Ng, Kamela Charmaine S.; Rivera, Windell L.

    2014-01-01

    Due to frequent antibiotic exposure, swine is now recognized as potential risk in disseminating drug-resistant Salmonella enterica strains. This study thus subjected 20 randomly selected S. enterica isolates from tonsil and jejunum with lymph node (JLN) tissues of swine slaughtered in Metro Manila, Philippines, to VITEK 2 antimicrobial susceptibility testing (AST). The test revealed all 20 isolates had resistance to at least one antimicrobial agent, in which highest occurrence of resistance was to amikacin (100%), cefazolin (100%), cefuroxime (100%), cefuroxime axetil (100%), cefoxitin (100%), and gentamicin (100%), followed by ampicillin (50%), and then by sulfamethoxazole trimethoprim (30%). Three multidrug-resistant (MDR) isolates were detected. The sole S. enterica serotype Enteritidis isolate showed resistance to 12 different antibiotics including ceftazidime, ceftriaxone, amikacin, gentamicin, and tigecycline. This study is the first to report worldwide on the novel resistance to tigecycline of MDR S. enterica serotype Enteritidis isolated from swine tonsil tissues. This finding poses huge therapeutic challenge since MDR S. enterica infections are associated with increased rate of hospitalization or death. Thus, continual regulation of antimicrobial use in food animals and prediction of resistant serotypes are crucial to limit the spread of MDR S. enterica isolates among hogs and humans. PMID:24724034

  20. First report of iliacus abscess caused by Salmonella enterica serovar Othmarschen.

    PubMed

    Jha, Babita; Kim, Choon-Mee; Kim, Dong-Min; Chung, Jong-Hoon; Yoon, Na-Ra; Jha, Piyush; Kim, Seok Won; Jang, Sook Jin; Kim, Seon Gyeong; Chung, Jae Keun

    2016-02-01

    The non-typhoidal bacterium Salmonella enterica subspecies enterica serovar Othmarschen (Salmonella Othmarschen) is a rare human pathogen. Abscess formation due to non-typhoidal Salmonella infections is a very rare complication in this antibiotic era. We report the first case of iliacus abscess after a short period of gastroenteritis which was caused by non-typhoidal Salmonella enterica belonging to group C1, serovar Othmarschen in a patient without any underlying conditions. A young female presented in our hospital complaining of pain in right hip joint area. She gave a history of watery diarrhea 3 days before the onset of pain. On examination the patient was ill-looking and there was tenderness in the right hip joint area. S. enterica was identified using 16S rRNA gene amplification by PCR and serotyped to be serovar Othmarschen from the pus sample of iliacus abscess. This is the first reported case of iliacus abscess due to Salmonella serover Othmarschen infection. Our case suggests that S. enterica serovar Othmarschen can cause severe focal infections associated with gastroenteritis. The literature on the rare association of Salmonella enterica and abscess formation is reviewed. PMID:26482919

  1. Fate of Salmonella enterica in a mixed ingredient salad containing lettuce, cheddar cheese, and cooked chicken meat.

    PubMed

    Bovo, Federica; De Cesare, Alessandra; Manfreda, Gerardo; Bach, Susan; Delaquis, Pascal

    2015-03-01

    Food service and retail sectors offer consumers a variety of mixed ingredient salads that contain fresh-cut vegetables and other ingredients such as fruits, nuts, cereals, dairy products, cooked seafood, cooked meat, cured meats, or dairy products obtained from external suppliers. Little is known about the behavior of enteric bacterial pathogens in mixed ingredient salads. A model system was developed to examine the fate of Salmonella enterica (inoculum consisting of S. enterica serovars Agona, Typhimurium, Enteritidis, Brandenberg, and Kentucky) on the surface of romaine lettuce tissues incubated alone and in direct contact with Cheddar cheese or cooked chicken. S. enterica survived but did not grow on lettuce tissues incubated alone or in contact with Cheddar cheese for 6 days at either 6 or 14°C. In contrast, populations increased from 2.01 ± 0.22 to 9.26 ± 0.22 CFU/cm(2) when lettuce washed in water was incubated in contact with cooked chicken at 14°C. Populations on lettuce leaves were reduced to 1.28 ± 0.14 CFU/cm(2) by washing with a chlorine solution (70 ppm of free chlorine) but increased to 8.45 ± 0.22 CFU/cm(2) after 6 days at 14°C. Experimentation with a commercial product in which one third of the fresh-cut romaine lettuce was replaced with inoculated lettuce revealed that S. enterica populations increased by 4 log CFU/g during storage for 3 days at 14°C. These findings indicate that rapid growth of bacterial enteric pathogens may occur in mixed ingredient salads; therefore, strict temperature control during the manufacture, distribution, handling, and storage of these products is critical. PMID:25719871

  2. Salmonella enterica Burden in Harvest-Ready Cattle Populations from the Southern High Plains of the United States▿

    PubMed Central

    Kunze, David J.; Loneragan, Guy H.; Platt, Tammy M.; Miller, Mark F.; Besser, Thomas E.; Koohmaraie, Mohammad; Stephens, Tyler; Brashears, Mindy M.

    2008-01-01

    Our objectives were to quantify the Salmonella enterica burdens in harvest-ready cattle and to identify specific at-risk populations of cattle most likely to harbor multiply resistant S. enterica. Hide swabs were collected in abattoirs from three cohorts of cattle (feedlot origin cattle that had achieved desirable harvest characteristics and dairy- and beef-type cows harvested because of poor productivity). Feces were collected from two cohorts housed in feedlots (cattle that had achieved desirable harvest characteristics and animals identified for salvage recovery because of poor productivity). Facilities were visited on four occasions over a 12-month period. Salmonella enterica isolates were recovered, and organisms were quantified using standard microbiological methodologies. Susceptibility to antimicrobial drugs and serotype were determined for one S. enterica isolate per sample. Salmonella enterica was recovered from 55.6% of 1,681 samples. The prevalences on hides and in feces were 69.6% and 30.3%, respectively. The concentrations of S. enterica organisms averaged (as determined by the most probable number technique) 1.82 log10/100 cm2 of hides and 0.75 log10/g of feces. None of the isolates recovered from cattle that had achieved desirable harvest characteristics were resistant to four or more drugs. For isolates recovered from animals with poor productivity characteristics, 6.5% were resistant to four or more drugs. Twenty-two serovars were identified, with the most common being Salmonella enterica serovar Anatum (25.5%), Salmonella enterica serovar Montevideo (22.2%), and Salmonella enterica serovar Cerro (12.5%). High-level resistance, i.e., resistance to four or more drugs, was clustered within a few relatively uncommon serovars. These results demonstrate that even though S. enterica isolates are readily recoverable from harvest-ready cattle, multiply resistant variants are rare and are associated with specific serovars in cattle harvested because of

  3. Inflammatory Responses to Salmonella Infections Are Serotype-Specific

    PubMed Central

    Ktsoyan, Zhanna; Ghazaryan, Karine; Manukyan, Gayane; Martirosyan, Anush; Mnatsakanyan, Armine; Arakelova, Karine; Gevorgyan, Zaruhi; Sedrakyan, Anahit; Asoyan, Ara; Boyajyan, Anna; Aminov, Rustam

    2013-01-01

    The main purpose of this study was to investigate the profile of inflammatory response in patients with acute salmonellosis caused by two serotypes of Salmonella enterica, S. Enteritidis and S. Typhimurium, as well as in convalescent patients with previous acute disease caused by S. Enteritidis. Patients with acute disease showed significantly elevated levels of IL-1β, IL-17, IL-10, and calprotectin compared to healthy control subjects. In convalescent patients, these markers were also significantly elevated, with the exception of IL-1β. Multivariate statistical analyses with the use of these variables produced models with a good predictive accuracy resulting in excellent separation of the diseased and healthy cohorts studied. Overall, the results suggest that the profile of inflammatory response in this disease is determined, to a significant degree, by the serotype of Salmonella, and the profile of certain cytokines and calprotectin remains abnormal for a number of months following the acute disease stage. PMID:26904722

  4. Inflammatory Responses to Salmonella Infections Are Serotype-Specific.

    PubMed

    Ktsoyan, Zhanna; Ghazaryan, Karine; Manukyan, Gayane; Martirosyan, Anush; Mnatsakanyan, Armine; Arakelova, Karine; Gevorgyan, Zaruhi; Sedrakyan, Anahit; Asoyan, Ara; Boyajyan, Anna; Aminov, Rustam

    2013-01-01

    The main purpose of this study was to investigate the profile of inflammatory response in patients with acute salmonellosis caused by two serotypes of Salmonella enterica, S. Enteritidis and S. Typhimurium, as well as in convalescent patients with previous acute disease caused by S. Enteritidis. Patients with acute disease showed significantly elevated levels of IL-1β, IL-17, IL-10, and calprotectin compared to healthy control subjects. In convalescent patients, these markers were also significantly elevated, with the exception of IL-1β. Multivariate statistical analyses with the use of these variables produced models with a good predictive accuracy resulting in excellent separation of the diseased and healthy cohorts studied. Overall, the results suggest that the profile of inflammatory response in this disease is determined, to a significant degree, by the serotype of Salmonella, and the profile of certain cytokines and calprotectin remains abnormal for a number of months following the acute disease stage. PMID:26904722

  5. Population structure of Salmonella enterica subspecies enterica (subspecies 1)

    Technology Transfer Automated Retrieval System (TEKTRAN)

    We sequenced and assembled 354 new Salmonella enterica ssp. enterica genomes. These genomes were chosen to maximize genetic diversity, representing at least 100 different serovars and distinct PFGE patterns within these serovars. 119 of the strains were of known antibiotic resistance,...

  6. Substructure within Salmonella enterica subsp. enterica Isolates from Australian Wildlife▿

    PubMed Central

    Parsons, Sandra K.; Bull, C. Michael; Gordon, David M.

    2011-01-01

    Multilocus sequence typing of 56 Salmonella enterica subsp. enterica strains isolated from Australian wildlife hosts was performed. The results of population assignment algorithms revealed that the 56 strains could be subdivided into two distinct clades. Strains belonging to the two clades were further distinguished phenotypically, genotypically, and with respect to host distribution. PMID:21378038

  7. Subtyping of Salmonella Food Isolates Suggests the Geographic Clustering of Serotype Telaviv.

    PubMed

    Durul, Bora; Acar, Sinem; Bulut, Ece; Kyere, Emmanuel O; Soyer, Yeşim

    2015-12-01

    Salmonella is commonly found in a variety of food products and is a major cause of bacterial foodborne illness throughout the world. In this study, we investigated the prevalence and diversity of Salmonella in eight different food types: sheep ground meat, cow ground meat, chicken meat, cow offal, traditional Sanliurfa cheese, unripened feta cheese, pistachios, and isot (a spice blend of dried red peppers specific to Sanliurfa), traditionally and commonly consumed in Turkey. Among 192 food samples, Salmonella was detected in 59 samples, with the highest prevalence in raw poultry parts (58%) and offal (58%) samples, while Salmonella was not detected in pistachios and dried red pepper. Resultant Salmonella isolates were characterized by serotyping, multilocus sequence typing (MLST), and pulsed-field gel electrophoresis (PFGE). Ten different serotypes represented 10 MLST sequence types (STs) with 1 novel ST and 17 PFGE types. Antimicrobial resistance profiling revealed that 30.5% of the isolates were resistant to two or more antimicrobials. Salmonella enterica subsp. enterica serotype Telaviv, which is rare throughout the world, was the second most common serotype isolated from food samples in this study, suggesting that this serotype might be one of the subtypes that is endemic to Turkey. PMID:26489049

  8. Draft Genome Sequences of Salmonella enterica subsp. enterica Serovars Typhimurium and Nottingham Isolated from Food Products

    PubMed Central

    Zheng, Jie; Ayers, Sherry; Melka, David C.; Curry, Phillip E.; Payne, Justin S.; Laasri, Anna; Wang, Charles; Hammack, Thomas S.; Brown, Eric W.

    2016-01-01

    A quantitative real-time PCR (qPCR) designed to detect Salmonella enterica subsp. enterica serovar Enteritidis, targeting the sdf gene, generated positive results for S. enterica subsp. enterica serovar Typhimurium (CFSAN033950) and S. enterica subsp. enterica serovar Nottingham (CFSAN006803) isolated from food samples. Both strains show pulsed-field gel electrophoresis (PFGE) patterns distinct from those of S. Enteritidis. Here, we report the genome sequences of these two strains. PMID:27445384

  9. Characterisation of antimicrobial resistance-associated integrons and mismatch repair gene mutations in Salmonella serotypes.

    PubMed

    Yang, Baowei; Zheng, Jie; Brown, Eric W; Zhao, Shaohua; Meng, Jianghong

    2009-02-01

    In this study, we examined the presence of integrons and Salmonella genomic island 1 (SGI1) and assessed their contribution to antimicrobial resistance as well as determining the extent of the mutator phenotype in Salmonella isolates. A total of 81 Salmonella enterica serotype Typhimurium isolates were examined for the presence of integrons and SGI1 and for hypermutators using polymerase chain reaction (PCR) and the mutator assay, respectively. An additional 336 Salmonella isolates were also used to screen for hypermutators. Fourteen S. Typhimurium isolates carried class 1 integrons, of which six were shown to possess SGI1. Five putative mutators, S. Typhimurium ST20751, S. enterica serotype Heidelberg 22396 and S. enterica serotype Enteritidis 17929, 17929N and 17929R, were identified among the 417 Salmonella isolates. Complementation analysis with the wild-type mutH, mutL, mutS and uvrD genes indicated that none of the five mutators contained defective mismatch repair (MMR) system alleles. DNA sequence analysis revealed that single point mutations resulting in aspartic acid (codon 87) substitution in the gyrA gene conferred resistance to nalidixic acid and/or other fluoroquinolone drugs (ciprofloxacin and enrofloxacin) among four isolates. Our findings indicated that integrons and SGI1 play an important role in multidrug resistance in Salmonella. The incidence of hypermutators owing to defective MMR in Salmonella appears to be rare. PMID:19013057

  10. Validation and Identification of Invasive Salmonella Serotypes in Sub-Saharan Africa by Multiplex Polymerase Chain Reaction.

    PubMed

    Al-Emran, Hassan M; Krumkamp, Ralf; Dekker, Denise Myriam; Eibach, Daniel; Aaby, Peter; Adu-Sarkodie, Yaw; Ali, Mohammad; Rubach, Mathew P; Bjerregaard-Andersen, Morten; Crump, John A; Cruz Espinoza, Ligia Maria; Løfberg, Sandra Valborg; Gassama Sow, Amy; Hertz, Julian T; Im, Justin; Jaeger, Anna; Kabore, Leon Parfait; Konings, Frank; Meyer, Christian G; Niang, Aissatou; Pak, Gi Deok; Panzner, Ursula; Park, Se Eun; Rabezanahary, Henintsoa; Rakotozandrindrainy, Raphaël; Raminosoa, Tiana Mirana; Razafindrabe, Tsiriniaina Jean Luco; Sampo, Emmanuel; Schütt-Gerowitt, Heidi; Sarpong, Nimako; Soura, Abdramane Bassiahi; Tall, Adama; von Kalckreuth, Vera; Wierzba, Thomas F; May, Jürgen; Marks, Florian

    2016-03-15

    Salmonella enterica serovar Typhi and nontyphoidal Salmonella (NTS) cause the majority of bloodstream infections in sub-Saharan Africa; however, serotyping is rarely performed. We validated a multiplex polymerase chain reaction (PCR) assay with the White-Kauffmann-Le Minor (WKLM) scheme of serotyping using 110 Salmonella isolates from blood cultures of febrile children in Ghana and applied the method in other Typhoid Fever Surveillance in Africa Program study sites. In Ghana, 47 (43%) S. Typhi, 36 (33%) Salmonella enterica serovar Typhimurium, 14 (13%) Salmonella enterica serovar Dublin, and 13 (12%) Salmonella enterica serovar Enteritidis were identified by both multiplex PCR and the WKLM scheme separately. Using the validated multiplex PCR assay, we identified 42 (66%) S. Typhi, 14 (22%) S. Typhimurium, 2 (3%) S. Dublin, 2 (3%) S. Enteritidis, and 4 (6%) other Salmonella species from the febrile patients in Burkina Faso, Guinea-Bissau, Madagascar, Senegal, and Tanzania. Application of this multiplex PCR assay in sub-Saharan Africa could advance the knowledge of serotype distribution of Salmonella. PMID:26933026

  11. Genomic Variation of Salmonella enterica Serovar Paratyphi B (Tartrate +) Isolates from Clinical and Zoonotic Specimens as Measured by PFGE and Fragment Analysis of Multiplex PCR Products

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Many serovars of Salmonella enterica are important human and animal gastrointestinal pathogens that can result in morbidity and mortality. We developed a simple, quick, and sensitive molecular assay using a 12 target multiplex PCR assay to serotype multiple samples. In developing this 1 day assay we...

  12. Diversity of multidrug-resistant salmonella enterica strains associated with cattle at harvest in the United States.

    PubMed

    Brichta-Harhay, Dayna M; Arthur, Terrance M; Bosilevac, Joseph M; Kalchayanand, Norasak; Shackelford, Steven D; Wheeler, Tommy L; Koohmaraie, Mohammad

    2011-03-01

    The prevalence and diversity of multidrug-resistant (MDR) Salmonella enterica strains associated with cattle at harvest in the United States were examined. Hides and carcasses of cattle were sampled at processing plants (n = 6) located in four geographically distant regions from July 2005 to April 2006. The mean prevalences of Salmonella on hides, preevisceration carcasses (immediately after hide removal), and postintervention carcasses (in the chiller and after the full complement of interventions) were 89.6%, 50.2%, and 0.8%, respectively. The values for MDR Salmonella enterica strains (defined as those resistant to two or more antimicrobials) as percentages of Salmonella prevalence were 16.7% (95% confidence interval [CI], 8.3 to 25.1%; median percent prevalence, 6.9%), 11.7% (95% CI, 4.4 to 19.0%; median, 4.8%), and 0.33% (95% CI, -0.3 to 0.70%; median, 0%), respectively. In this study, 16,218 Salmonella hide and carcass isolates were screened for antimicrobial resistance. Of these, 978 (6.0%) unique MDR S. enterica isolates were identified and serotyped and their XbaI pulsed-field gel electrophoresis (PFGE) profiles determined. The predominant MDR S. enterica serotypes observed were Newport (53.1%), Typhimurium (16.6%), and Uganda (10.9%). Differences in MDR S. enterica prevalence were detected, and PFGE analysis revealed both epidemic clusters (profiles found in plants in multiple regions/seasons) and endemic clusters (profiles observed in plants in limited regions/seasons) within several of the MDR serotypes examined. Despite these differences, multiple-hurdle processing interventions employed at all plants were found to be quite effective and decreased Salmonella carcass contamination by 98.4% (95% CI, 97.6 to 99.7%). PMID:21239549

  13. Diversity of Multidrug-Resistant Salmonella enterica Strains Associated with Cattle at Harvest in the United States ▿

    PubMed Central

    Brichta-Harhay, Dayna M.; Arthur, Terrance M.; Bosilevac, Joseph M.; Kalchayanand, Norasak; Shackelford, Steven D.; Wheeler, Tommy L.; Koohmaraie , Mohammad

    2011-01-01

    The prevalence and diversity of multidrug-resistant (MDR) Salmonella enterica strains associated with cattle at harvest in the United States were examined. Hides and carcasses of cattle were sampled at processing plants (n = 6) located in four geographically distant regions from July 2005 to April 2006. The mean prevalences of Salmonella on hides, preevisceration carcasses (immediately after hide removal), and postintervention carcasses (in the chiller and after the full complement of interventions) were 89.6%, 50.2%, and 0.8%, respectively. The values for MDR Salmonella enterica strains (defined as those resistant to two or more antimicrobials) as percentages of Salmonella prevalence were 16.7% (95% confidence interval [CI], 8.3 to 25.1%; median percent prevalence, 6.9%), 11.7% (95% CI, 4.4 to 19.0%; median, 4.8%), and 0.33% (95% CI, −0.3 to 0.70%; median, 0%), respectively. In this study, 16,218 Salmonella hide and carcass isolates were screened for antimicrobial resistance. Of these, 978 (6.0%) unique MDR S. enterica isolates were identified and serotyped and their XbaI pulsed-field gel electrophoresis (PFGE) profiles determined. The predominant MDR S. enterica serotypes observed were Newport (53.1%), Typhimurium (16.6%), and Uganda (10.9%). Differences in MDR S. enterica prevalence were detected, and PFGE analysis revealed both epidemic clusters (profiles found in plants in multiple regions/seasons) and endemic clusters (profiles observed in plants in limited regions/seasons) within several of the MDR serotypes examined. Despite these differences, multiple-hurdle processing interventions employed at all plants were found to be quite effective and decreased Salmonella carcass contamination by 98.4% (95% CI, 97.6 to 99.7%). PMID:21239549

  14. Salmonella enterica bacteraemia: a multi-national population-based cohort study

    PubMed Central

    2010-01-01

    Background Salmonella enterica is an important emerging cause of invasive infections worldwide. However, population-based data are limited. The objective of this study was to define the occurrence of S. enterica bacteremia in a large international population and to evaluate temporal and regional differences. Methods We conducted population-based laboratory surveillance for all salmonella bacteremias in six regions (annual population at risk 7.7 million residents) in Finland, Australia, Denmark, and Canada during 2000-2007. Results A total of 622 cases were identified for an annual incidence of 1.02 per 100,000 population. The incidence of typhoidal (serotypes Typhi and Paratyphi) and non-typhoidal (other serotypes) disease was 0.21 and 0.81 per 100,000/year. There was major regional and moderate seasonal and year to year variability with an increased incidence observed in the latter years of the study related principally to increasing rates of non-typhoidal salmonella bacteremias. Advancing age and male gender were significant risk factors for acquiring non-typhoidal salmonella bacteremia. In contrast, typhoidal salmonella bacteremia showed a decreasing incidence with advancing age and no gender-related excess risk. Conclusions Salmonella enterica is an important emerging pathogen and regional determinants of risk merits further investigation. PMID:20398281

  15. Salmonella enterica resistant to antimicrobials in wastewater effluents and black-headed gulls in the Czech Republic, 2012.

    PubMed

    Masarikova, Martina; Manga, Ivan; Cizek, Alois; Dolejska, Monika; Oravcova, Veronika; Myskova, Petra; Karpiskova, Renata; Literak, Ivan

    2016-01-15

    We investigated the presence and epidemiological relatedness of Salmonella isolates from a wastewater treatment plant (WWTP) in Brno, Czech Republic and from nestlings of black-headed gulls (Chroicocephalus ridibundus) at the Nove Mlyny waterworks, situated 35 km downstream from the WWTP. During 2012, we collected 37 wastewater samples and 284 gull cloacal swabs. From wastewater samples, we obtained 89 Salmonella isolates belonging to 19 serotypes. At least one resistant strain was contained in 89% of those samples. Ten different serotypes of Salmonella were detected in 38 young gulls, among which 14 (37%) were resistant to antimicrobials. Wastewater isolates were mostly resistant to sulphonamides and tetracycline, gull isolates to tetracycline and ampicillin. We detected the occurrence of blaTEM-1,tet(A), tet(B), tet(G), sul1, sul2, sul3, floR and strA resistance genes. For the first time, we identified a class 1 integron with the dfrA12-orfF-aadA2 gene cassette in S. Infantis. Using pulsed-field gel electrophoresis, we confirmed the presence of identical clusters of S. Agona, S. Enteritidis PT8, S. Infantis and S. Senftenberg in wastewater and black-headed gulls, thus indicating the possibility of resistant Salmonella isolates spreading over long distances in the environment. PMID:26519571

  16. A serotype 10 human rotavirus.

    PubMed

    Beards, G; Xu, L; Ballard, A; Desselberger, U; McCrae, M A

    1992-06-01

    Rotaviruses with genome rearrangements isolated from a chronically infected immunodeficient child (F. Hundley, M. McIntyre, B. Clark, G. Beards, D. Wood, I. Chrystie, and U. Desselberger, J. Virol 61:3365-3372, 1987) are the first recognized human isolates of serotype 10. This was shown by both a direct enzyme-linked immunosorbent assay and virus neutralization assays using serotype specific monoclonal antibodies. The serotype was confirmed by sequence analysis of the gene encoding VP7, which revealed a 96% amino acid homology to the bovine serotype 10 isolate B223. PMID:1320627

  17. Involvement of a Salmonella Genomic Island 1 Gene in the Rumen Protozoan-Mediated Enhancement of Invasion for Multiple-Antibiotic-Resistant Salmonella enterica Serovar Typhimurium▿

    PubMed Central

    Carlson, Steve A.; Sharma, Vijay K.; McCuddin, Zoe P.; Rasmussen, Mark A.; Franklin, Sharon K.

    2007-01-01

    Multiple-antibiotic-resistant Salmonella enterica serotype Typhimurium is a food-borne pathogen that may be more virulent than related strains lacking the multiresistance phenotype. Salmonella enterica serotype Typhimurium phage type DT104 is the most prevalent of these multiresistant/hypervirulent strains. Multiresistance in DT104 is conferred by an integron structure, designated Salmonella genomic island 1 (SGI1), while we recently demonstrated DT104 hyperinvasion mediated by rumen protozoa (RPz) that are normal flora of cattle. Hyperinvasion was also observed in other Salmonella strains, i.e., other S. enterica serovar Typhimurium phage types and other S. enterica serovars, like S. enterica serovar Infantis, possessing SGI1, while DT104 strains lacking SGI1 were not hyperinvasive. Herein we attempted to identify SGI1 genes involved in the RPz-mediated hyperinvasion of Salmonella strains bearing SGI1. Transposon mutagenesis, coupled with a novel reporter system, revealed the involvement of an SGI1 gene previously designated SO13. Disruption of SO13 expression led to an abrogation of hyperinvasion as assessed by tissue culture invasion assays and by bovine challenge experiments. However, hyperinvasion was not observed in non-SGI1-bearing strains of Salmonella engineered to express SO13. That is, SO13 and another SGI1 gene(s) may coordinately upregulate invasion in DT104 exposed to RPz. PMID:17145942

  18. Integrative Analysis of Salmonellosis in Israel Reveals Association of Salmonella enterica Serovar 9,12:l,v:− with Extraintestinal Infections, Dissemination of Endemic S. enterica Serovar Typhimurium DT104 Biotypes, and Severe Underreporting of Outbreaks

    PubMed Central

    Marzel, Alex; Desai, Prerak T.; Nissan, Israel; Schorr, Yosef Ilan; Suez, Jotham; Valinsky, Lea; Reisfeld, Abraham; Agmon, Vered; Guard, Jean; McClelland, Michael

    2014-01-01

    Salmonella enterica is the leading etiologic agent of bacterial food-borne outbreaks worldwide. This ubiquitous species contains more than 2,600 serovars that may differ in their host specificity, clinical manifestations, and epidemiology. To characterize salmonellosis epidemiology in Israel and to study the association of nontyphoidal Salmonella (NTS) serovars with invasive infections, 48,345 Salmonella cases reported and serotyped at the National Salmonella Reference Center between 1995 and 2012 were analyzed. A quasi-Poisson regression was used to identify irregular clusters of illness, and pulsed-field gel electrophoresis in conjunction with whole-genome sequencing was applied to molecularly characterize strains of interest. Three hundred twenty-nine human salmonellosis clusters were identified, representing an annual average of 23 (95% confidence interval [CI], 20 to 26) potential outbreaks. We show that the previously unsequenced S. enterica serovar 9,12:l,v:− belongs to the B clade of Salmonella enterica subspecies enterica, and we show its frequent association with extraintestinal infections, compared to other NTS serovars. Furthermore, we identified the dissemination of two prevalent Salmonella enterica serovar Typhimurium DT104 clones in Israel, which are genetically distinct from other global DT104 isolates. Accumulatively, these findings indicate a severe underreporting of Salmonella outbreaks in Israel and provide insights into the epidemiology and genomics of prevalent serovars, responsible for recurring illness. PMID:24719441

  19. The taxonomic structure of Salmonella enterica subspecies enterica

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Salmonella enterica is the leading cause of food-borne bacterial infection in humans and has a high economic burden in agriculture. Strains differ by sequence additions and losses of up to ~10% of each genome. In the last few decades, some serovars have become more common. Many strains have acquired...

  20. Revisitingmolecular serotyping of Streptococcus pneumoniae

    PubMed Central

    2015-01-01

    Background Ninety-two Streptococcus pneumoniae serotypes have been described so far, but the pneumococcal conjugate vaccine introduced in the Brazilian basic vaccination schedule in 2010 covers only the ten most prevalent in the country. Pneumococcal serotype-shifting after massive immunization is a major concern and monitoring this phenomenon requires efficient and accessible serotyping methods. Pneumococcal serotyping based on antisera produced in animals is laborious and restricted to a few reference laboratories. Alternatively, molecular serotyping methods assess polymorphisms in the cps gene cluster, which encodes key enzymes for capsular polysaccharides synthesis in pneumococci. In one such approach, cps-RFLP, the PCR amplified cps loci are digested with an endonuclease, generating serotype-specific fingerprints on agarose gel electrophoresis. Methods In this work, in silico and in vitro approaches were combined to demonstrate that XhoII is the most discriminating endonuclease for cps-RFLP, and to build a database of serotype-specific fingerprints that accommodates the genetic diversity within the cps locus of 92 known pneumococci serotypes. Results The expected specificity of cps-RFLP using XhoII was 76% for serotyping and 100% for serogrouping. The database of cps-RFLP fingerprints was integrated to Molecular Serotyping Tool (MST), a previously published web-based software for molecular serotyping. In addition, 43 isolates representing 29 serotypes prevalent in the state of Minas Gerais, Brazil, from 2007 to 2013, were examined in vitro; 11 serotypes (nine serogroups) matched the respective in silico patterns calculated for reference strains. The remaining experimental patterns, despite their resemblance to their expected in silico patterns, did not reach the threshold of similarity score to be considered a match and were then added to the database. Conclusion The cps-RFLP method with XhoII outperformed the antisera-based and other molecular serotyping

  1. Microarray-Based Detection of Salmonella enterica Serovar Enteritidis Genes Involved in Chicken Reproductive Tract Colonization

    PubMed Central

    Raspoet, R.; Appia-Ayme, C.; Shearer, N.; Martel, A.; Pasmans, F.; Haesebrouck, F.; Ducatelle, R.; Thompson, A.

    2014-01-01

    Salmonella enterica serovar Enteritidis has developed the potential to contaminate table eggs internally, by colonization of the chicken reproductive tract and internalization in the forming egg. The serotype Enteritidis has developed mechanisms to colonize the chicken oviduct more successfully than other serotypes. Until now, the strategies exploited by Salmonella Enteritidis to do so have remained largely unknown. For that reason, a microarray-based transposon library screen was used to identify genes that are essential for the persistence of Salmonella Enteritidis inside primary chicken oviduct gland cells in vitro and inside the reproductive tract in vivo. A total of 81 genes with a potential role in persistence in both the oviduct cells and the oviduct tissue were identified. Major groups of importance include the Salmonella pathogenicity islands 1 and 2, genes involved in stress responses, cell wall, and lipopolysaccharide structure, and the region-of-difference genomic islands 9, 21, and 40. PMID:25281378

  2. Isolation of Salmonella enterica and serologic reactivity to Leptospira interrogans in opossums (Didelphis virginiana) from Yucatán, México.

    PubMed

    Ruiz-Pina, Hugo Antonio; Puc-Franco, Miguel Angel; Flores-Abuxapqui, Javier; Vado-Solis, Ignacio; Cardenas-Marrufo, María Fidelia

    2002-01-01

    The presence of Salmonella enterica and serologic evidence of infection by Leptospira interrogans, were detected in the opossum Didelphis virginiana in a semi-urban locality of the Yucatán State, México. Ninety-one opossums were captured during the period April 1996 and May 1998. From a total of 17 feces samples, four Salmonella enterica subsp. enterica serotypes (Sandiego, Newport, Anatum, and Minnesota), and one Salmonella enterica subsp. arizonae serovar O44:Z4,Z23:- were isolated. Some opossums presented mixed infections. From 81 sera samples, four (4.9%) were positive to antibodies to Leptospira serovars pomona and wolfii. Both animals infected with Salmonella enterica and those serologically positive to Leptospira interrogans were captured in peridomestic habitat. Opossums infected with Salmonella enterica, were captured in dry season, and those seropositive to Leptospira interrogans during the rainy season. The implications of infection and reactivity of these zoonotic pathogens in D. virginiana in the Yucatan state are briefly discussed. PMID:12219118

  3. High-throughput molecular determination of salmonella enterica serovars by use of multiplex PCR and capillary electrophoresis analysis.

    PubMed

    Leader, Brandon T; Frye, Jonathan G; Hu, Jinxin; Fedorka-Cray, Paula J; Boyle, David S

    2009-05-01

    Salmonella enterica is a leading cause of food-borne illness worldwide and is also a major cause of morbidity and mortality in domestic and wild animals. In the current study, a high-throughput molecular assay was developed to determine the most common clinical and nonhuman serovars of S. enterica in the United States. Sixteen genomic targets were identified based on their differential distribution among common serovars. Primers were designed to amplify regions of each of these targets in a single multiplex PCR while incorporating a 6-carboxyfluorescein-labeled universal primer to fluorescently label all amplicons. The fluorescently labeled PCR products were separated using capillary electrophoresis, and a Salmonella multiplex assay for rapid typing (SMART) code was generated for each isolate, based upon the presence or absence of PCR products generated from each target gene. Seven hundred fifty-one blind clinical isolates of Salmonella from Washington State, collected in 2007 and previously serotyped via antisera, were screened with the assay. A total of 89.6% of the isolates were correctly identified based on comparison to a panel of representative SMART codes previously determined for the top 50 most common serovars in the United States. Of the remaining isolates, 6.2% represented isolates that produced a new SMART code for a previously determined serotype, while the final 8.8% were from serotypes not screened in the original panel used to score isolates in the blinded study. This high-throughput multiplex PCR assay allowed simple and accurate typing of the most prevalent clinical serovars of Salmonella enterica at a level comparable to that of conventional serotyping, but at a fraction of both the cost and time required per test. PMID:19261787

  4. Structural background for serological cross-reactivity between bacteria of different enterobacterial serotypes.

    PubMed

    Makszin, Lilla; Péterfi, Zoltán; Blaskó, Ágnes; Sándor, Viktor; Kilár, Anikó; Dörnyei, Ágnes; Ősz, Erzsébet; Kilár, Ferenc; Kocsis, Béla

    2015-06-01

    The structure of the oligosaccharide repeating units of endotoxins from Gram-negative bacteria is characteristic for the different serogroups and serotypes of bacteria. Detailed examination of the cross-reactions of three enterobacterial serotypes, Proteus morganii O34, Escherichia coli O111, and Salmonella enterica sv. Adelaide O35, was performed using sensitive tests (ELISA, immunoblotting). Fine differences between the endotoxins of the bacteria were detected using silver staining of SDS-PAGE gels and chip-technology for the intact lipopolysaccharides (LPSs). The compositions of the O-specific polysaccharides of LPSs extracted from the bacteria were studied, and it was proven that the three cross-reacting bacteria contain O-antigens built from the same monosaccharides, namely colitoses linked to glucose, galactose, and N-acetyl-galactosamine. The NMR and GC-MS studies revealed that the most probable component for the cross-reaction is the rare sugar, colitose. PMID:25630395

  5. Serotype IX, a Proposed New Streptococcus agalactiae Serotype.

    PubMed

    Slotved, Hans-Christian; Kong, Fanrong; Lambertsen, Lotte; Sauer, Susanne; Gilbert, Gwendolyn L

    2007-09-01

    We identified three isolates of Streptococcus agalactiae (group B streptococcus [GBS]), of human origin, which failed to react with antisera against any of the nine known GBS serotypes. Polyclonal rabbit antisera raised against these isolates and standard GBS typing sera were used in capillary precipitation and Ouchterlony tests to compare the strains with known GBS serotype reference strains. All three previously nontypeable isolates reacted with all three new antisera, producing lines of identity in the Ouchterlony test. Weak cross-reactions with antisera against several GBS serotypes were observed but were removed by absorption with corresponding antigens. The new antisera were used to test 227 GBS isolates that had been nontypeable or difficult to type using standard antisera. Of these, five reacted with the new antisera. These results suggested that all eight isolates belong to the previously unrecognized GBS serotype. They were tested by Western blotting for the Calpha and Cbeta proteins and by PCR to identify molecular serotypes and surface protein antigen genes. Two segments of the cps gene cluster (3' end of cpsE-cpsF and 5' end of cpsG, approximately 700 bp; 3' end of cpsH and 5' end of cpsM, approximately 560 bp) were sequenced. All eight isolates expressed Calpha, and seven expressing the Cbeta protein and the corresponding genes, bca and bac, respectively, were identified. They all share the same, unique partial cps sequence. These results indicate that these eight isolates represent a new S. agalactiae serotype, which we propose should be designated serotype IX. PMID:17634306

  6. Recovery rates, serotypes, and antimicrobial susceptibility patterns of salmonellae isolated from cloacal swabs of wild Nile crocodiles (Crocodylus niloticus) in Zimbabwe.

    PubMed

    Madsen, M; Hangartner, P; West, K; Kelly, P

    1998-03-01

    Samples from the cloaca and the ventral skin surface of 67 Nile crocodiles (Crocodylus niloticus) captured in four uninhabited areas at Lake Kariba, Zimbabwe, were cultured for Salmonella. All the skin samples tested negative for Salmonella, whereas 18 of 67 (26.9%) cloacal samples grew Salmonella. Significantly more males than females yielded Salmonella, but no statistically significant correlation among salmonella carriage, body size, and age was recorded. Ten different serotypes of S. enterica belonging to the subspecies enterica, salamae, and diarizonae were isolated. All isolates belonging to subspecies enterica displayed invasive properties in an experimental mouse model and thus exhibited pathogenic potential, whereas none of the other isolates were invasive. In general, isolates were sensitive to a number of commonly used antimicrobials, except for three isolates that were resistant to streptomycin. PMID:9638622

  7. Complete Genome and Methylome Sequences of Two Salmonella enterica spp.

    PubMed Central

    Yao, Kuan; Muruvanda, Tim; Roberts, Richard J.; Payne, Justin; Allard, Marc W.

    2016-01-01

    Salmonella enterica is responsible for major foodborne outbreaks worldwide. It can cause gastroenteritis characterized by diarrhea, vomiting, and fever. Salmonella infections raise public health concerns along with consequential economic impacts. In this report, we announce the first complete genome sequences of Salmonella enterica subsp. enterica serovar Choleraeuis (S. Choleraeuis) ATCC 10708 and Salmonella enterica subsp. enterica serovar Pullorum (S. Pullorum) ATCC 9120, isolated from patients with diarrhea. PMID:26798102

  8. Effect of desiccation on tolerance of salmonella enterica to multiple stresses.

    PubMed

    Gruzdev, Nadia; Pinto, Riky; Sela, Shlomo

    2011-03-01

    Reducing the available water in food is a long-established method for controlling bacterial growth in the food industry. Nevertheless, food-borne outbreaks of salmonellosis due to consumption of dry foods have been continuously reported. Previous studies showed that dried Salmonella cells acquire high tolerance to heat and ethanol. In order to examine if dehydration also induces tolerance to other stressors, dried Salmonella enterica serotype Typhimurium cells were exposed to multiple stresses, and their viability was assessed. Indeed, desiccated S. Typhimurium acquired higher tolerance to multiple stressors than nondesiccated cells. The dried cells were significantly more resistant to most stressors, including ethanol (10 to 30%, 5 min), sodium hypochlorite (10 to 100 ppm, 10 min), didecyl dimethyl ammonium chloride (0.05 to 0.25%, 5 min), hydrogen peroxide (0.5 to 2.0%, 30 min), NaCl (0.1 to 1 M, 2 h), bile salts (1 to 10%, 2 h), dry heat (100°C, 1 h), and UV irradiation (125 μW/cm(2), 25 min). In contrast, exposure of Salmonella to acetic and citric acids reduced the survival of the dried cells (1.5 log) compared to that of nondesiccated cells (0.5 log). Three other S. enterica serotypes, S. Enteritidis, S. Newport, and S. Infantis, had similar stress responses as S. Typhimurium, while S. Hadar was much more susceptible and gained tolerance to only a few stressors. Our findings indicate that dehydration induces cross-tolerance to multiple stresses in S. enterica, demonstrating the limitations of current chemical and physical treatments utilized by the food industry to inactivate food-borne pathogens. PMID:21216905

  9. Prevalence, Distribution, and Diversity of Salmonella enterica in a Major Produce Region of California▿†

    PubMed Central

    Gorski, Lisa; Parker, Craig T.; Liang, Anita; Cooley, Michael B.; Jay-Russell, Michele T.; Gordus, Andrew G.; Atwill, E. Robert; Mandrell, Robert E.

    2011-01-01

    A survey was initiated to determine the prevalence of Salmonella enterica in the environment in and around Monterey County, CA, a major agriculture region of the United States. Trypticase soy broth enrichment cultures of samples of soil/sediment (n = 617), water (n = 252), wildlife (n = 476), cattle feces (n = 795), and preharvest lettuce and spinach (n = 261) tested originally for the presence of pathogenic Escherichia coli were kept in frozen storage and later used to test for the presence of S. enterica. A multipathogen oligonucleotide microarray was employed to identify a subset of samples that might contain Salmonella in order to test various culture methods to survey a larger number of samples. Fifty-five of 2,401 (2.3%) samples yielded Salmonella, representing samples obtained from 20 different locations in Monterey and San Benito Counties. Water had the highest percentage of positives (7.1%) among sample types. Wildlife yielded 20 positive samples, the highest number among sample types, with positive samples from birds (n = 105), coyotes (n = 40), deer (n = 104), elk (n = 39), wild pig (n = 41), and skunk (n = 13). Only 16 (2.6%) of the soil/sediment samples tested positive, and none of the produce samples had detectable Salmonella. Sixteen different serotypes were identified among the isolates, including S. enterica serotypes Give, Typhimurium, Montevideo, and Infantis. Fifty-four strains were sensitive to 12 tested antibiotics; one S. Montevideo strain was resistant to streptomycin and gentamicin. Pulsed-field gel electrophoresis (PFGE) analysis of the isolates revealed over 40 different pulsotypes. Several strains were isolated from water, wildlife, or soil over a period of several months, suggesting that they were persistent in this environment. PMID:21378057

  10. [Classify species of Pseudomonas pseudomallei into serotypes].

    PubMed

    Han, O; Li, L; Zhao, Z

    1990-10-01

    Species of P. pseudomallei can be classified into two serotypes, serotype I and serotype II, based on whether or not it contains a thermolabile antigen beside a thermostable one. Under the condition of lack of typing serum, by means of serum absorption test, we recognized a strain which contains a major thermolabile antigen. The antigen was purified by Sephadex G-200, and it was used to inoculate rabbits. With the immunoserum at hand, we identified 68 of the domestic chinese strains and 6 of alien strains for serotyping by bi-directional agar diffusion test. The results showed that 68 strains were identified serotype I, 3 strains serotype II, and the remaining 3 comparable with those reported indicating that strains of serotype I were found mostly in Asia, and that the serotype are unrelated to their existing environments, nor that of animal bodies, but connected with their existence in geographic distribution. PMID:2251832

  11. Potential of Three-Way Randomly Amplified Polymorphic DNA Analysis as a Typing Method for Twelve Salmonella Serotypes

    PubMed Central

    Soto, S. M.; Guerra, B.; González-Hevia, M. A.; Mendoza, M. C.

    1999-01-01

    The potential of a three-way randomly amplified polymorphic DNA (RAPD) procedure (RAPD typing) for typing Salmonella enterica strains assigned to 12 serotypes was analyzed. The series of organisms used included 235 strains (326 isolates) collected mainly from clinical samples in the Principality of Asturias and 9 reference strains. RAPD typing was performed directly with broth cultures of bacteria by using three selected primers and optimized PCR conditions. The profiles obtained with the three primers were used to define RAPD types and to evaluate the procedure as a typing method at the species and serotype levels. The typeability was 100%; the reproducibility and in vitro stability could be considered good. The concordance of RAPD typing methods with serotyping methods was 100%, but some profiles obtained with two of the three primers were obtained with strains assigned to different serotypes. The discrimination index (DI) within the series of organisms was 0.94, and the DI within serotypes Typhimurium, Enteritidis, and Virchow were 0.72, 0.52, and 0.66, respectively. Within these serotypes the most common RAPD types were differentiated into phage types and vice versa; combining the types identified by the two procedures (RAPD typing and phage typing) resulted in further discrimination (DI, 0.96, 0.74, and 0.87, respectively). The efficiency, rapidity, and flexibility of the RAPD typing method support the conclusion that it can be used as a tool for identifying Salmonella organisms and as a typing method that is complementary to serotyping and phage typing methods. PMID:10543793

  12. Cross-sectional Study Examining Salmonella enterica Carriage in Subiliac Lymph Nodes of Cull and Feedlot Cattle at Harvest

    PubMed Central

    Gragg, Sara E.; Loneragan, Guy H.; Brashears, Mindy M.; Arthur, Terrance M.; Bosilevac, Joseph M.; Kalchayanand, Norasak; Wang, Rong; Schmidt, John W.; Brooks, J. Chance; Shackelford, Steven D.; Wheeler, Tommy L.; Brown, Tyson R.; Edrington, Thomas S.

    2013-01-01

    Abstract Bovine peripheral lymph nodes (LNs), including subiliac LNs, have been identified as a potential source of human exposure to Salmonella enterica, when adipose trim containing these nodes is incorporated into ground beef. In order to gain a better understanding of the burden of S. enterica in peripheral LNs of feedlot and cull cattle, a cross-sectional study was undertaken in which 3327 subiliac LNs were collected from cattle at harvest in seven plants, located in three geographically distinct regions of the United States. Samples were collected in three seasons: Fall 2010, Winter/Spring 2011, and Summer/Fall 2011. A convenience sample of 76 LNs per day, 2 days per season (approximately 1 month apart), was collected per plant, from carcasses held in the cooler for no less than 24 h. Every 10th carcass half on a rail was sampled, in an attempt to avoid oversampling any single cohort of cattle. Median point estimates of S. enterica contamination were generally low (1.3%); however, median Salmonella prevalence was found to be greater in subiliac LNs of feedlot cattle (11.8%) compared to those of cull cattle (0.65%). Enumeration analysis of a subset of 618 feedlot cattle LNs showed that 67% of those harboring S. enterica (97 of 144) did so at concentrations ranging from <0.1 to 1.8 log10 CFU/g, while 33% carried a higher burden of S. enterica, with levels ranging from 1.9 to >3.8 log10 CFU/g. Serotyping of S. enterica isolated identified 24 serotypes, with the majority being Montevideo (44.0%) and Anatum (24.8%). Antimicrobial susceptibility phenotypes were determined for all isolates, and the majority (86.1%) were pansusceptible; however, multidrug-resistant isolates (8.3%) were also occasionally observed. As Salmonella contained within LNs are protected from carcass interventions, research is needed to define opportunities for mitigating the risk of Salmonella contamination in LNs of apparently healthy cattle. PMID:23566273

  13. Antimicrobial susceptibilities of isolates of Staphylococcus aureus, Listeria species and Salmonella serotypes associated with poultry processing.

    PubMed

    Geornaras, I; von Holy, A

    2001-10-22

    The broth microdilution method was used to determine the activities of selected antimicrobial agents used in the South African poultry industry (danofloxacin, neomycin, chlortetracycline, oxytetracycline, tylosin and colistin) and vancomycin against bacterial isolates previously obtained from carcasses and selected equipment surfaces and environmental sources associated with poultry processing. The antimicrobial susceptibilities of 38 isolates of Staphylococcus (S.) aureus, 25 Listeria (L.) innocua, 18 L. monocytogenes, and 62 isolates belonging to six Salmonella (Salm.) serotypes (Salm. agona, Salm. blockley, Salm. enteritidis, Salm. isangi, Salm. reading and Salm. typhimurium) were determined. The most active antimicrobial agent against all the isolates tested was danofloxacin with minimum inhibitory concentrations (MICs) for 90% of the isolates (MIC90) not exceeding 0.25 and 2 microg/ml for gram-negative and gram-positive isolates, respectively. Conversely, high MICs were recorded for all the isolates tested against chlortetracycline and oxytetracycline (MIC90 range of 32 to > 512 microg/ml), except for the L. monocytogenes and Salm. enteritidis isolates (MIC range of < or = 0.5-4 microg/ml). Neomycin was found to be active against S. aureus, L. innocua, L. monocytogenes, Salm. enteritidis and Salm. isangi isolates, with MICs not exceeding 8 microg/ml. MIC ranges for tylosin and vancomycin, which were only tested against the gram-positive isolates, were from 1 to > 512 microg/ml and from 1 to 4 microg/ml, respectively. The MIC range for the remaining antimicrobial agent, colistin, which was only tested against the Salmonella isolates, was 0.5-16 microg/ml. The lack of MIC breakpoints for the antimicrobial agents used in the poultry industry did not allow for definite conclusions as to the level of resistant bacteria associated with poultry carcasses and the processing environment in this study. PMID:11759760

  14. Antimicrobial resistance of Salmonella enterica isolates from bulk tank milk and milk filters in the United States.

    PubMed

    Van Kessel, J S; Sonnier, J; Zhao, S; Karns, J S

    2013-01-01

    Salmonella isolates were recovered from bulk tank milk as part of the National Animal Health Monitoring System (NAHMS) Dairy 2002 and 2007 surveys. In-line milk filters were also tested in the 2007 survey. The objective of this study was to determine the prevalence of antimicrobial resistance among Salmonella enterica isolates from bulk milk and milk filters in the NAHMS Dairy 2002 and 2007 surveys and to further characterize resistant isolates. Susceptibilities to 15 antibiotics were determined for 176 Salmonella isolates of 26 serotypes using an automated antimicrobial susceptibility system. Resistant isolates were screened by PCR for the presence of the extended-spectrum β-lactamase (bla(CMY)) gene and class I integrons and further characterized by pulsed-field gel electrophoresis. Thirty isolates (17.0%) representing six S. enterica serotypes exhibited resistance to at least one antimicrobial agent (serotypes Newport [14 of 14 isolates exhibited resistance], Dublin [7 of 7], Typhimurium [3 of 5], Kentucky [4 of 22], Anatum [1 of 13], and Infantis [1 of 2]). Twenty isolates (11.4%), including all 14 Newport, 3 Dublin, 2 Typhimurium, and 1 Infantis isolate, displayed the typical multidrug-resistant, bla(CMY)-positive (MDR-AmpC) phenotype which included resistance to ampicillin, chloramphenicol, streptomycin, sulfonamide, and tetracycline, plus resistance to amoxicillin-clavulanic acid and extended-spectrum cephalosporins. Five of the MDR-AmpC isolates carried class I integrons (2.8%). Two-enzyme (XbaI and BlnI) pulsed-field gel electrophoresis discerned clades within serotypes and, together with the resistance profiles, identified strains that appeared to have persisted temporally and geographically. These results suggest that there is a low but appreciable risk of infection with MDR Salmonella from consumption of nonpasteurized milk and dairy products. PMID:23317852

  15. Whole-genome sequencing of Salmonella enterica subsp. enterica serovar Cubana strains isolated from agricultural sources

    Technology Transfer Automated Retrieval System (TEKTRAN)

    We report draft genomes of Salmonella enterica subsp. enterica Serovar Cubana strain CVM42234 isolated from chick feed in 2012 and Salmonella Cubana strain 76814 isolated from swine in 2004. The genome sizes are 4,975,046 and 4,936,251 base pairs, respectively....

  16. Complete Genome Sequence and Methylome of Salmonella enterica subsp. enterica Cerro, a Frequent Dairy Cow Serovar

    PubMed Central

    Haley, Bradd J.; Pirone, Cary; Muruvanda, Tim; Brown, Eric; Allard, Marc; Karns, Jeffrey S.

    2016-01-01

    Salmonella enterica subsp. enterica serovar Cerro is an infrequent pathogen of humans and other mammals but is frequently isolated from the hindgut of asymptomatic cattle in the United States. To further understand the genomic determinants of S. Cerro specificity for the bovine hindgut, the genome of isolate CFSAN001588 was fully sequenced and deposited in the GenBank database. PMID:26823571

  17. Whole-Genome Sequencing of Salmonella enterica subsp. enterica Serovar Ouakam Isolated from Ground Turkey

    PubMed Central

    Marasini, Daya; Abo-Shama, Usama H.

    2016-01-01

    In this report, we announce the first whole-genome sequencing of Salmonella enterica subsp. enterica serovar Ouakam strain GNT-01, isolated from ground turkey retail meat. The strain has a chromosome of 5,088,451 bp long, with a G+C content of 52.3%, and a plasmid of 109,715 bp. PMID:26798110

  18. Draft Genome Sequence of Salmonella enterica subsp. enterica Serovar Mishmarhaemek Isolated from Bovine Feces

    PubMed Central

    Cooper, Ashley; Lambert, Dominic; Koziol, Adam G.; Seyer, Karine

    2015-01-01

    Salmonella enterica subsp. enterica serovar Mishmarhaemek is a Gram-negative, non-spore-forming, rod-shaped bacterium implicated in human clinical disease. Here, we report a 4.8-Mbp draft genome sequence of a nalidixic acid-resistant isolate of S. serovar Mishmarhaemek. PMID:26472847

  19. Complete genome sequence of salmonella enterica subsp. enterica Serovar Thompson Strain RM6836

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Salmonella enterica subsp. enterica serovar Thompson (S. Thompson) strain RM6836 was isolated from lettuce in 2002. We report the complete sequence and annotation of the genome of S. Thompson strain RM6836. This is the first reported complete genome sequence for S. Thompson and will provide a point ...

  20. Salmonella enterica Genomics and Antimicrobial Resistance

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Salmonella is a prevalent food-borne pathogen and a model system for the study of virulence and pathogenesis. The development of DNA microarray technology has furthered investigation of genome organization that leads to the variations in Salmonella serotypes. There are over 2400 Salmonella serotypes...

  1. Complete and Closed Genome Sequences of 10 Salmonella enterica subsp. enterica Serovar Anatum Isolates from Human and Bovine Sources

    PubMed Central

    Nguyen, Scott V.; Bono, James L.; Smith, Timothy P. L.; Fields, Patricia I.; Dinsmore, Blake A.; Santovenia, Monica; Kelley, Christy M.; Wang, Rong; Bosilevac, Joseph M.; Harhay, Gregory P.

    2016-01-01

    Salmonella enterica is an important pathogen transmitted by numerous vectors. Genomic comparisons of Salmonella strains from disparate hosts have the potential to further our understanding of mechanisms underlying host specificities and virulence. Here, we present the closed genome and plasmid sequences of 10 Salmonella enterica subsp. enterica serovar Anatum isolates from bovine and human sources. PMID:27257192

  2. Complete and Closed Genome Sequences of 10 Salmonella enterica subsp. enterica Serovar Anatum Isolates from Human and Bovine Sources.

    PubMed

    Nguyen, Scott V; Harhay, Dayna M; Bono, James L; Smith, Timothy P L; Fields, Patricia I; Dinsmore, Blake A; Santovenia, Monica; Kelley, Christy M; Wang, Rong; Bosilevac, Joseph M; Harhay, Gregory P

    2016-01-01

    Salmonella enterica is an important pathogen transmitted by numerous vectors. Genomic comparisons of Salmonella strains from disparate hosts have the potential to further our understanding of mechanisms underlying host specificities and virulence. Here, we present the closed genome and plasmid sequences of 10 Salmonella enterica subsp. enterica serovar Anatum isolates from bovine and human sources. PMID:27257192

  3. Complete Genome Sequences of Two Outbreak Strains of Salmonella enterica subsp. enterica Serovar Thompson Associated with Cilantro

    PubMed Central

    Huynh, Steven; Gorski, Lisa; Cooper, Kerry K.; Miller, William G.

    2015-01-01

    Salmonella enterica subsp. enterica serovar Thompson strains RM1984 (CADPH-99A2334) and RM1986 (CADPH-99A2345) are associated with a 1999 outbreak in contaminated cilantro. We report here the complete genome sequences and annotation of these two S. Thompson strains. These genomes are distinct and provide additional data for our understanding of S. enterica. PMID:26586897

  4. Complete Genome Sequences of Two Outbreak Strains of Salmonella enterica subsp. enterica Serovar Thompson Associated with Cilantro.

    PubMed

    Parker, Craig T; Huynh, Steven; Gorski, Lisa; Cooper, Kerry K; Miller, William G

    2015-01-01

    Salmonella enterica subsp. enterica serovar Thompson strains RM1984 (CADPH-99A2334) and RM1986 (CADPH-99A2345) are associated with a 1999 outbreak in contaminated cilantro. We report here the complete genome sequences and annotation of these two S. Thompson strains. These genomes are distinct and provide additional data for our understanding of S. enterica. PMID:26586897

  5. Evolutionary Genomics of Salmonella enterica Subspecies.

    PubMed

    Desai, Prerak T; Porwollik, Steffen; Long, Fred; Cheng, Pui; Wollam, Aye; Bhonagiri-Palsikar, Veena; Hallsworth-Pepin, Kymberlie; Clifton, Sandra W; Weinstock, George M; McClelland, Michael

    2013-01-01

    ABSTRACT Six subspecies are currently recognized in Salmonella enterica. Subspecies I (subspecies enterica) is responsible for nearly all infections in humans and warm-blooded animals, while five other subspecies are isolated principally from cold-blooded animals. We sequenced 21 phylogenetically diverse strains, including two representatives from each of the previously unsequenced five subspecies and 11 diverse new strains from S. enterica subspecies enterica, to put this species into an evolutionary perspective. The phylogeny of the subspecies was partly obscured by abundant recombination events between lineages and a relatively short period of time within which subspeciation took place. Nevertheless, a variety of different tree-building methods gave congruent evolutionary tree topologies for subspeciation. A total of 285 gene families were identified that were recruited into subspecies enterica, and most of these are of unknown function. At least 2,807 gene families were identified in one or more of the other subspecies that are not found in subspecies I or Salmonella bongori. Among these gene families were 13 new candidate effectors and 7 new candidate fimbrial clusters. A third complete type III secretion system not present in subspecies enterica (I) isolates was found in both strains of subspecies salamae (II). Some gene families had complex taxonomies, such as the type VI secretion systems, which were recruited from four different lineages in five of six subspecies. Analysis of nonsynonymous-to-synonymous substitution rates indicated that the more-recently acquired regions in S. enterica are undergoing faster fixation rates than the rest of the genome. Recently acquired AT-rich regions, which often encode virulence functions, are under ongoing selection to maintain their high AT content. IMPORTANCE We have sequenced 21 new genomes which encompass the phylogenetic diversity of Salmonella, including strains of the previously unsequenced subspecies arizonae

  6. Serotype assignment by sero-agglutination, ELISA, and PCR

    Technology Transfer Automated Retrieval System (TEKTRAN)

    For assessing isolates of Listeria monocytogenes serotype designation is the foremost subtyping method used. Traditionally serotyping has been done with agglutination reactions. In the last decade alternative serotyping methods were described using Enzyme Linked Immunosorbent Assay(ELISA)and Polymer...

  7. Development and Evaluation of a Multiplex Real-Time Polymerase Chain Reaction Procedure to Clinically Type Prevalent Salmonella enterica Serovars

    PubMed Central

    Muñoz, Nélida; Diaz-Osorio, Miguel; Moreno, Jaime; Sánchez-Jiménez, Miryan; Cardona-Castro, Nora

    2010-01-01

    A multiplex real-time polymerase chain reaction procedure was developed to identify the most prevalent clinical isolates of Salmonella enterica subsp. enterica. Genes from the rfb, fliC, fljB, and viaB groups that encode the O, H, and Vi antigens were used to design 15 primer pairs and TaqMan probes specific for the genes rfbJ, wzx, fliC, fljB, wcdB, the sdf-l sequence, and invA, which was used as an internal amplification control. The primers and probes were variously combined into six sets. The first round of reactions used two of these sets to detect Salmonella O:4, O:9, O:7, O:8, and O:3,10 serogroups. Once the serogroups were identified, the results of a second round of reactions that used primers and probes for the flagellar antigen l genes, 1,2; e,h; g,m; d; e,n,x; and z10, and the Vi gene were used to identify individual serovars. The procedure was standardized using 18 Salmonella reference strains and other enterobacteria. The procedure's reliability and sensitivity was evaluated using 267 randomly chosen serotyped Salmonella clinical isolates. The procedure had a sensitivity of 95.5% and was 100% specific. Thus, our technique is a quick, sensitive, reliable, and specific means of identifying S. enterica serovars and can be used in conjunction with traditional serotyping. Other primer and probe combinations could be used to increase the number of identifiable serovars. PMID:20110454

  8. Assessment of antibiotic resistance phenotype and integrons in Salmonella enterica serovar Typhimurium isolated from swine.

    PubMed

    Rayamajhi, Nabin; Kang, Sang Gyun; Kang, Mi Lan; Lee, Hee Soo; Park, Kyung Yoon; Yoo, Han Sang

    2008-10-01

    Salmonella enterica serovar Typhimurium (S. Typhimurium) isolated and identified from swine were subjected for the analysis of antibiotic resistance pattern and clinically important class 1 and 2 integrons. In addition, S. Typhimurium isolates exhibiting ampicillin, chloramphenicol, streptomycin, sulfamethoxazole, tetracycline and florfenicol (ACSSuTF) resistance pattern as described in most Salmonella enterica serotype Typhimurium definitive type 104 (DT104) were characterized by polymerase chain reaction. All the isolates were resistant to more than four antibiotics and showed the highest resistance to streptomycin (94.1%), followed by tetracycline (90.1%), ampicillin (64.7%), chloramphenicol (56.8%) and gentamicin (54.9%). MIC value for the ten isolates ranged between 0.125-2 mug/ml for ciprofloxacin. Among the beta-lactams used, only one of the isolate exhibited resistance to ceftiofur (MIC 8 microg/ml). Sixty eight percent of these multi drug resistance (MDR) S. Typhimurium isolates carried clinically important class 1 integron with 1kb (aadA) and/or 2kb (dhfrXII-orfF-aadA2) resistance gene cassettes. This study reports the increasing trend of multi drug resistance (MDR) S. Typhimurium with clinically important class 1 integron in pigs. In addition, emergence of the ACSSuTF-type resistance in S. Typhimurium PT other than DT104 may limit the use of resistance gene markers in its detection methods by PCR. PMID:18981675

  9. Reduced fluoroquinolone susceptibility in Salmonella enterica serotypes in travelers returning from Southeast Asia.

    PubMed

    Hakanen, A; Kotilainen, P; Huovinen, P; Helenius, H; Siitonen, A

    2001-01-01

    During 1995 to 1999, we collected 1,210 Salmonella isolates; 629 were from Finnish travelers returning from abroad. These isolates were tested for susceptibility by determining MICs to ciprofloxacin, nalidixic acid, and seven additional antimicrobial agents. From 1995 to 1999, the annual proportion of reduced ciprofloxacin susceptibility (MIC > 0.125 microg/mL) among all travelers' isolates increased from 3.9% to 23.5% (p<0.001). The increasing trend was outstanding among the isolates from Southeast Asia; isolates from Thailand alone increased from 5.6% to 50.0% (p<0.001). The reduced fluoroquinolone susceptibility was nonclonal in character and significantly associated with multidrug resistance. A point mutation in the quinolone resistance-determining region of gyrA was present in all isolates with reduced susceptibility. These data provide further evidence for the rapid spread of multidrug-resistant pathogens from one continent to another. PMID:11747728

  10. Incidence and growth of Salmonella enterica on the peel and pulp of avocado (Persea americana) and custard apple (Annona squamosa).

    PubMed

    Rezende, Ana Carolina B; Crucello, Juliana; Moreira, Rafael C; Silva, Beatriz S; Sant'Ana, Anderson S

    2016-10-17

    The aim of this study was to assess the incidence and to estimate the growth kinetic parameters (maximum growth rate, μ; lag time, λ; and maximum population, κ) of Salmonella on the peel and pulp of avocado (Perseaamericana var. americana) and custard apple (Annona squamosa L.) as affected by temperature (10-30°C). The incidence of Salmonella was assessed on the peel and pulp of the fruits (n=200 of each fruit), separately, totalizing 800 analyses. Only three samples of custard apple pulp were positive for Salmonella enterica and the three isolates recovered belonged to serotype S. Typhimurium. Salmonella was not recovered from avocado and custard apple peels and from avocado pulp. Generally, the substrate (pulp or peel) of growth did not affect μ values of S. enterica (p>0.05). Very similar μ values were found for S. enterica inoculated in custard apple and avocado. S. enterica presented the highest λ in the peel of the fruits. The growth of S. enterica resulted in larger λ in custard apple in comparison to avocado. For example, the λ of S. enterica in the pulp of custard apple and avocado were 47.0±0.78h and 10.0±3.78h, respectively. The lowest values of κ were obtained at the lower storage temperature conditions (10°C). For instance, κ values of 3.7±0.06log CFU/g and 2.9±0.03log CFU/g were obtained from the growth of S. enterica in avocado and custard apple pulps at 10°C (p<0.05), respectively. On the other hand, at 30°C, κ values were 6.5±0.25log CFU/g and 6.5±0.05log CFU/g, respectively. Significantly higher κ were obtained from the growth of S. enterica in the pulp than in the peel of the fruits (p<0.05). For instance, the growth of S. enterica in the pulp of avocado led to a κ value of 6.5±0.25log CFU/g, while in the peel led to a κ value of 4.6±0.23log CFU/g (p<0.05). In general, growth kinetic parameters indicated that avocado comprises a better substrate than custard apple for the growth of S. enterica. The square root model

  11. A serotype-specific polymerase chain reaction for identification of Pasteurella multocida serotype 1

    USGS Publications Warehouse

    Rocke, Tonie E.; Smith, Susan R.; Miyamoto, Amy; Shadduck, Daniel J.

    2002-01-01

    A serotype-specific polymerase chain reaction (PCR) assay was developed for detection and identification of Pasteurella multocida serotype 1, the causative agent of avian cholera in wild waterfowl. Arbitrarily primed PCR was used to detect DNA fragments that distinguish serotype 1 from the other 15 serotypes of P. multocida (with the exception of serotype 14). Oligonucleotide primers were constructed from these sequences, and a PCR assay was optimized and evaluated. PCR reactions consistently resulted in amplification products with reference strains 1 and 14 and all other serotype 1 strains tested, with cell numbers as low as 2.3 cells/ml. No amplification products were produced with other P. multocida serotypes or any other bacterial species tested. To compare the sensitivity and further test the specificity of this PCR assay with traditional culturing and serotyping techniques, tissue samples from 84 Pekin ducks inoculated with field strains of P. multocida and 54 wild lesser snow geese collected during an avian cholera outbreak were provided by other investigators working on avian cholera. PCR was as sensitive (58/64) as routine isolation (52/64) in detecting and identifying P. multocida serotype 1 from the livers of inoculated Pekins that became sick or died from avian cholera. No product was amplified from tissues of 20 other Pekin ducks that received serotypes other than type 1 (serotype 3, 12 × 3, or 10) or 12 control birds. Of the 54 snow geese necropsied and tested for P. multocida, our PCR detected and identified the bacteria from 44 compared with 45 by direct isolation. The serotype-specific PCR we developed was much faster and less labor intensive than traditional culturing and serotyping procedures and could result in diagnosis of serotype 1 pasteurellosis within 24 hr of specimen submission.

  12. A serotype-specific polymerase chain reaction for identification of Pasteurella multocida serotype 1

    USGS Publications Warehouse

    Rocke, T.E.; Smith, S.R.; Miyamoto, A.; Shadduck, D.J.

    2002-01-01

    A serotype-specific polymerase chain reaction (PCR) assay was developed for detection and identification of Pasteurella multocida serotype 1, the causative agent of avian cholera in wild waterfowl. Arbitrarily primed PCR was used to detect DNA fragments that distinguish serotype 1 from the other 15 serotypes of P. multocida (with the exception of serotype 14). Oligonucleotide primers were constructed from these sequences, and a PCR assay was optimized and evaluated. PCR reactions consistently resulted in amplification products with reference strains 1 and 14 and all other serotype 1 strains tested, with cell numbers as low as 2.3 cells/ml. No amplification products were produced with other P. multocida serotypes or any other bacterial species tested. To compare the sensitivity and further test the specificity of this PCR assay with traditional culturing and serotyping techniques, tissue samples from 84 Pekin ducks inoculated with field strains of P. multocida and 54 wild lesser snow geese collected during an avian cholera outbreak were provided by other investigators working on avian cholera. PCR was as sensitive (58/64) as routine isolation (52/64) in detecting and identifying P. multocida serotype 1 from the livers of inoculated Pekins that became sick or died from avian cholera. No product was amplified from tissues of 20 other Pekin ducks that received serotypes other than type 1 (serotype 3, 12 ?? 3, or 10) or 12 control birds. Of the 54 snow geese necropsied and tested for P. multocida, our PCR detected and identified the bacteria from 44 compared with 45 by direct isolation. The serotype-specific PCR we developed was much faster and less labor intensive than traditional culturing and serotyping procedures and could result in diagnosis of serotype 1 pasteurellosis within 24 hr of specimen submission.

  13. Two Draft Genome Sequences of a New Serovar of Salmonella enterica, Serovar Lubbock

    PubMed Central

    den Bakker, Henk C.; Nightingale, Kendra K.; Brichta-Harhay, Dayna M.; Edrington, Thomas S.; Loneragan, Guy H.

    2015-01-01

    Salmonella enterica is principally a foodborne pathogen that shows considerable serovar diversity. In this report, we present two draft genome sequences of Salmonella enterica subsp. enterica serovar Lubbock, a novel serovar. PMID:25883279

  14. Two draft genome sequences of a new serovar of Salmonella enterica, serovar Lubbock

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Salmonella enterica is principally a foodborne pathogen that shows considerable serovar diversity. In this report, we present two draft genome sequences of Salmonella enterica subsp. enterica serovar Lubbock, a novel serovar....

  15. Detection of Salmonella enterica in pigs at slaughter and comparison with human isolates in Italy.

    PubMed

    Bonardi, Silvia; Alpigiani, Irene; Bruini, Ilaria; Barilli, Elena; Brindani, Franco; Morganti, Marina; Cavallini, Pierugo; Bolzoni, Luca; Pongolini, Stefano

    2016-02-01

    In 2013-2014, 201 pigs belonging to 67 batches were tested for Salmonella in their mesenteric lymph nodes (MLN) in one abattoir of Northern Italy. For each batch, faecal material was collected at lairage by swabbing the pen floor for approximately 1600 cm(2). The aim of this study was to investigate the prevalence of Salmonella in MLN of pigs at slaughter, to assess Salmonella contamination at lairage and to evaluate the effect of lairage duration on its prevalence. Serotyping, XbaI PFGE typing and antimicrobial testing of the isolates were performed. Pig and human Salmonella isolates of the same region of Italy were compared to evaluate possible correlations. Salmonella enterica was isolated from 19.9% of the MLN and 49.3% of the environmental faecal samples. Nine different serovars were identified among 75 S. enterica isolates. In MLN Salmonella Derby was the most common (52.5%), followed by S. enterica 4,[5],12:i:- (17.5%) and Salmonella Rissen (10.0%). In faecal samples S. Derby was prevalent (51.4%), followed by S. enterica 4,[5], 12:i:- (20.0%) and Salmonella Brandenburg (14.3%). Lairage holding varied between 1 and ≥ 12 h (median value: 2.5h). In pigs held for 1-3h, 14.1% were positive for Salmonella in MLN but the prevalence reached 31.8% when they were held for ≥ 12 h. The contamination of MLN was statistically different (p=0.0045) between the two groups, thus confirming the role of long-lasting lairage in Salmonella contamination of pigs. XbaI PFGE typing detected 36 PFGE types. Twenty-three PFGE types were identified among the 40 MLN isolates and 22 PFGE types among the 35 faecal isolates. A total of 11 PFGE types were shared between the MLN of pigs and the lairage environment. Among S. Derby, 6 shared PFGE types between MLN and faeces were found and among S. enterica 4,[5],12:i:- one PFGE type was common between MLN and the faecal samples. Shared profiles between human and swine isolates of S. Derby, S. enterica 4,[5],12:i:-, S. Rissen, Salmonella

  16. Requirement for cobalamin by Salmonella enterica serovars Typhimurium, Pullorum, Gallinarum and Enteritidis during infection in chickens

    PubMed Central

    Vaz, Jacqueline Boldrin; Penha Filho, Rafael Antonio Casarin; Junior, Angelo Berchieri; Lemos, Manoel Victor Franco

    2011-01-01

    Salmonella enterica serovar Typhimurium synthesizes cobalamin (vitamin B12) only during anaerobiosis. Two percent of the S. Typhimurium genome is devoted to the synthesis and uptake of vitamin B12 and to B12-dependent reactions. To understand the requirement for cobalamin synthesis better, we constructed mutants of Salmonella serovars Enteritidis and Pullorum that are double-defective in cobalamin biosynthesis (ΔcobSΔcbiA). We compared the virulence of these mutants to that of their respective wild type strains and found no impairment in their ability to cause disease in chickens. We then assessed B12 production in these mutants and their respective wild type strains, as well as in S. Typhimurium ΔcobSΔcbiA, Salmonella Gallinarum ΔcobSΔcbiA, and their respective wild type strains. None of the mutants was able to produce detectable B12. B12 was detectable in S. Enteritidis, S. Pullorum and S. Typhimurium wild type strains but not in S. Gallinarum. In conclusion, the production of vitamin B12in vitro differed across the tested Salmonella serotypes and the deletion of the cbiA and cobS genes resulted in different levels of alteration in the host parasite interaction according to Salmonella serotype tested. PMID:24031771

  17. Pediatric Pneumococcal Serotypes in 4 European Countries

    PubMed Central

    Kissling, Esther; Fenoll, Asuncion; George, Robert; Lepoutre, Agnes; Lernout, Tinne; Tarragó, David; Varon, Emmanuelle; Verhaegen, Jan

    2010-01-01

    After heptavalent pneumococcal conjugate vaccine (PCV7) was marketed in France, Spain, Belgium, and England and Wales (United Kingdom), invasive disease from non-PCV7 serotypes (NVT) increased. Adjusted serotype-specific incidences among children <15 years of age were compared between 1999–2002 (prevaccine) and 2005–2006 (postmarketing). Vaccine coverage increased to ≈32%–48% in France, Spain, and Belgium but remained <1% in England and Wales. Serotype 1 incidence rose in all age groups and countries (incidence rate ratio [IRR] 1.3–4.2; p<0.004), independently of PCV7 use, but incidence of serotypes 7F and 19A increased most in France, Spain, and Belgium (IRR 1.9–16.9 in children <5 years; p<0.001), where PCV7 coverage was greater. Vaccine-induced replacement of PCV7 serotypes possibly contributed to NVT increases, as did secular trends. New vaccines targeting these serotypes are available, but serotype dynamics needs further exploration that accounts for underreporting and prevaccine trends. PMID:20735928

  18. Serotyping of Streptococcus pneumoniae Based on Capsular Genes Polymorphisms

    PubMed Central

    Raymond, Frédéric; Boucher, Nancy; Allary, Robin; Robitaille, Lynda; Lefebvre, Brigitte; Tremblay, Cécile

    2013-01-01

    Streptococcus pneumoniae serotype epidemiology is essential since serotype replacement is a concern when introducing new polysaccharide-conjugate vaccines. A novel PCR-based automated microarray assay was developed to assist in the tracking of the serotypes. Autolysin, pneumolysin and eight genes located in the capsular operon were amplified using multiplex PCR. This step was followed by a tagged fluorescent primer extension step targeting serotype-specific polymorphisms. The tagged primers were then hybridized to a microarray. Results were exported to an expert system to identify capsular serotypes. The assay was validated on 166 cultured S. pneumoniae samples from 63 different serotypes as determined by the Quellung method. We show that typing only 12 polymorphisms located in the capsular operon allows the identification at the serotype level of 22 serotypes and the assignation of 24 other serotypes to a subgroup of serotypes. Overall, 126 samples (75.9%) were correctly serotyped, 14 were assigned to a member of the same serogroup, 8 rare serotypes were erroneously serotyped, and 18 gave negative serotyping results. Most of the discrepancies involved rare serotypes or serotypes that are difficult to discriminate using a DNA-based approach, for example 6A and 6B. The assay was also tested on clinical specimens including 43 cerebrospinal fluid samples from patients with meningitis and 59 nasopharyngeal aspirates from bacterial pneumonia patients. Overall, 89% of specimens positive for pneumolysin were serotyped, demonstrating that this method does not require culture to serotype clinical specimens. The assay showed no cross-reactivity for 24 relevant bacterial species found in these types of samples. The limit of detection for serotyping and S. pneumoniae detection was 100 genome equivalent per reaction. This automated assay is amenable to clinical testing and does not require any culturing of the samples. The assay will be useful for the evaluation of serotype

  19. Muscle Abscess due to Salmonella Enterica

    PubMed Central

    Akkoyunlu, Yasemin; Ceylan, Bahadir; Iraz, Meryem; Elmadag, Nuh Mehmet; Aslan, Turan

    2013-01-01

    Non typhoidal Salmonellae spp. causes clinical symptoms especially in neonates, infants, aged and immunocompromised patients. Hematogenous dissemination may occur in complicated cases whereas the formation of abscess is rare. A 61-year old woman presented to our hospital with pain and a mass in her left arm, without fever and leukocytosis. She was using methotrexate, corticosteroids and quinine for rheumatoid arthritis. She had a history of cervix cancer and was given radiotherapy and chemotherapy 3 years ago. Upon physical examination and magnetic resonance imaging, the mass was considered as an abscess and was surgically drained. Salmonella enterica spp. enterica was yielded in the culture of the drainage material. Ceftriaxon 2g/day was started intramuscularly and continued for 4 weeks. Salmonellosis is usually a self-limited disease, generally restricted to gastrointestinal tract and acquired following food poisoning. Management of Salmonella abscess requires a combination of antibiotherapy, surgical drainage and eradication of primary foci. PMID:24396582

  20. Transmission and retention of Salmonella enterica by phytophagous hemipteran insects.

    PubMed

    Soto-Arias, José Pablo; Groves, Russell L; Barak, Jeri D

    2014-09-01

    Several pest insects of human and livestock habitations are known as vectors of Salmonella enterica; however, the role of plant-feeding insects as vectors of S. enterica to agricultural crops remains unexamined. Using a hemipteran insect pest-lettuce system, we investigated the potential for transmission and retention of S. enterica. Specifically, Macrosteles quadrilineatus and Myzus persicae insects were fed S. enterica-inoculated lettuce leaf discs or artificial liquid diets confined in Parafilm sachets to allow physical contact or exclusively oral ingestion of the pathogen, respectively. After a 24-h acquisition access period, insects were moved onto two consecutive noninoculated leaf discs or liquid diets and allowed a 24-h inoculation access period on each of the two discs or sachets. Similar proportions of individuals from both species ingested S. enterica after a 24-h acquisition access period from inoculated leaf discs, but a significantly higher proportion of M. quadrilineatus retained the pathogen internally after a 48-h inoculation access period. S. enterica was also recovered from the honeydew of both species. After a 48-h inoculation access period, bacteria were recovered from a significantly higher proportion of honeydew samples from M. quadrilineatus than from M. persicae insects. The recovery of S. enterica from leaf discs and liquid diets postfeeding demonstrated that both species of insects were capable of transmitting the bacteria in ways that are not limited to mechanical transmission. Overall, these results suggest that phytophagous insects may serve as potential vectors of S. enterica in association with plants. PMID:24973069

  1. Transmission and Retention of Salmonella enterica by Phytophagous Hemipteran Insects

    PubMed Central

    Soto-Arias, José Pablo; Groves, Russell L.

    2014-01-01

    Several pest insects of human and livestock habitations are known as vectors of Salmonella enterica; however, the role of plant-feeding insects as vectors of S. enterica to agricultural crops remains unexamined. Using a hemipteran insect pest-lettuce system, we investigated the potential for transmission and retention of S. enterica. Specifically, Macrosteles quadrilineatus and Myzus persicae insects were fed S. enterica-inoculated lettuce leaf discs or artificial liquid diets confined in Parafilm sachets to allow physical contact or exclusively oral ingestion of the pathogen, respectively. After a 24-h acquisition access period, insects were moved onto two consecutive noninoculated leaf discs or liquid diets and allowed a 24-h inoculation access period on each of the two discs or sachets. Similar proportions of individuals from both species ingested S. enterica after a 24-h acquisition access period from inoculated leaf discs, but a significantly higher proportion of M. quadrilineatus retained the pathogen internally after a 48-h inoculation access period. S. enterica was also recovered from the honeydew of both species. After a 48-h inoculation access period, bacteria were recovered from a significantly higher proportion of honeydew samples from M. quadrilineatus than from M. persicae insects. The recovery of S. enterica from leaf discs and liquid diets postfeeding demonstrated that both species of insects were capable of transmitting the bacteria in ways that are not limited to mechanical transmission. Overall, these results suggest that phytophagous insects may serve as potential vectors of S. enterica in association with plants. PMID:24973069

  2. Serotypes in Saccharomyces telluris: Their relation to source of isolation

    USGS Publications Warehouse

    Hasenclever, H.F.; Kocan, R.M.

    1973-01-01

    Three serotypes have been characterized with three reference strains of Saccharomyces telluris and designated as A, B, and C. One reference strain of Torpulopsis bovina, the imperfect form of S. telluris, belonged to serotype B. Strains of S. telluris isolated from four columbid species were serotyped. All 98 strains of this yeast isolated from Columba livia belonged to serotype B. Three other columbid species, C. leucocephala, C. fasciata, and Zenaidura macroura harbored strains of serotype C only. Serotype A was not isolated from any of the avian species.

  3. Phenotypic and Genotypic Characterization of Salmonella enterica in Captive Wildlife and Exotic Animal Species in Ohio, USA.

    PubMed

    Farias, L F P; Oliveira, C J B; Medardus, J J; Molla, B Z; Wolfe, B A; Gebreyes, W A

    2015-09-01

    The purpose of this study was to investigate the occurrence, antimicrobial resistance patterns, phenotypic and genotypic relatedness of Salmonella enterica recovered from captive wildlife host species and in the environment in Ohio, USA. A total of 319 samples including faecal (n = 225), feed (n = 38) and environmental (n = 56) were collected from 32 different wild and exotic animal species in captivity and their environment in Ohio. Salmonellae were isolated using conventional culture methods and tested for antimicrobial susceptibility with the Kirby-Bauer disc diffusion method. Salmonella isolates were serotyped, and genotyping was performed using the pulsed-field gel electrophoresis (PFGE). Salmonella was detected in 56 of 225 (24.9%) faecal samples; six of 56 (10.7%) environmental samples and six of 38 (15.8%) feed samples. Salmonella was more commonly isolated in faecal samples from giraffes (78.2%; 36/46), cranes (75%; 3/4) and raccoons (75%; 3/4). Salmonella enterica serotypes of known public health significance including S. Typhimurium (64.3%), S. Newport (32.1%) and S. Heidelberg (5.3%) were identified. While the majority of the Salmonella isolates were pan-susceptible (88.2%; 60 of 68), multidrug-resistant strains including penta-resistant type, AmStTeKmGm (8.8%; six of 68) were detected. Genotypic diversity was found among S. Typhimurium isolates. The identification of clonally related Salmonella isolates from environment and faeces suggests that indirect transmission of Salmonella among hosts via environmental contamination is an important concern to workers, visitors and other wildlife. Results of this study show the diversity of Salmonella serovars and public health implications of human exposure from wildlife reservoirs. PMID:25388917

  4. Complete genomic sequences of two outbreak strains of Salmonella enterica subsp. enterica serovar Thompson associated with cilantro

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Salmonella enterica subsp. enterica serovar Thompson strains RM1984 (CADPH-99A2334) and RM1986 (CADPH -99A2345) are clinical isolates from 1999, putatively related to an outbreak in California from contaminated cilantro. We report the complete genome sequences and annotation of these two S. Thompson...

  5. Genome Sequences of Salmonella enterica subsp. enterica Serovar Lubbock Strains Isolated from Liver Abscesses of Feedlot Cattle

    PubMed Central

    Amachawadi, Raghavendra G.; Thomas, Milton

    2016-01-01

    The genome sequencing of 13 Salmonella enterica subsp. enterica serovar Lubbock strains isolated from liver abscesses of feedlot cattle is reported here. The availability of these genomes will help to further understand the etiologic role of Salmonella strains in liver abscesses of cattle and will serve as references in microbial trace-back studies to improve food safety. PMID:27151794

  6. The complete genome sequence and methylome of Salmonella enterica subsp. enterica serovar Cerro, a frequent dairy cow strain

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Salmonella enterica subsp. enterica serovar Cerro is an infrequent pathogen of humans and other mammals, but is frequently isolated from the hindgut of asymptomatic cattle in the United States. To further understand the genomic determinants of S. Cerro specificity for the bovine hindgut, the genome ...

  7. Complete Genomic Sequences of Two Outbreak Strains of Salmonella enterica subsp. enterica serovar Thompson Associated with Cilantro

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Salmonella enterica subsp. enterica serovar Thompson strains RM1984 (CADPH-99A2334) and RM1986 (CADPH -99A2345) are clinical isolates from 1999, putatively related to an outbreak in California from contaminated cilantro. We report the complete genome sequences and annotation of these two S. Thompson...

  8. Diversity and antimicrobial susceptibility of Salmonella enterica serovars isolated from pig farms in Ibadan, Nigeria.

    PubMed

    Fashae, Kayode; Hendriksen, Rene S

    2014-01-01

    Animals including food animals play a significant role in the epidemiology of Salmonella enterica. The control requires identification of sources and institution of targeted interventions. This study investigates the diversity of S. enterica serovars, antimicrobial susceptibility, and occurrence of plasmid-mediated quinolone resistance (PMQR) genes in pigs in Ibadan, Nigeria. Pooled fresh pen floor fecal samples of pigs collected from 31 pig farms were cultured; the Salmonella isolates were serotyped and their antimicrobial susceptibility was determined. PMQR genes were screened by polymerase chain reaction. The 229 Salmonella isolates were made of 50 serovars predominated by rare serovars Salmonella Give (n = 36; 15.7 %), Salmonella Brancaster (n = 17; 7.4 %), Salmonella Colindale (n = 15; 6.6 %), Salmonella Elisaberthville (n = 13; 5.7 %), Salmonella Hillingdon (n = 13; 5.7 %), and Salmonella Kingston (n = 13; 5.7 %). The most widely distributed serovars among the farms were Salmonella Give (six farms) and Salmonella Elisaberthville (six farms). Resistance to chloramphenicol, sulfonamides, nalidixic acid, streptomycin, and tetracycline ranged from 11.6 % (n = 26) to 22.8 % (n = 51). Resistance ciprofloxacin and gentamicin was low (n = 2; 0.9 %). Multiply resistant isolates included Salmonella Kentucky, the most resistant serovar. qnrB19 was found in two isolates of Salmonella Corvallis and one isolate of Salmonella Larochelle, respectively, while qnrS1 was found in two isolates of Salmonella Derby. Other PMQR genes were not detected. Pigs constitute an important source of diverse Salmonella serovars in Ibadan. The isolates were more resistant to old antimicrobials with some multiple resistant. Control measures and regulation of antimicrobials are warranted. PMID:23893398

  9. Distribution and Characterization of Salmonella enterica Isolates from Irrigation Ponds in the Southeastern United States.

    PubMed

    Luo, Zhiyao; Gu, Ganyu; Ginn, Amber; Giurcanu, Mihai C; Adams, Paige; Vellidis, George; van Bruggen, Ariena H C; Danyluk, Michelle D; Wright, Anita C

    2015-07-01

    Irrigation water has been implicated as a likely source of produce contamination by Salmonella enterica. Therefore, the distribution of S. enterica was surveyed monthly in irrigation ponds (n = 10) located within a prime agricultural region in southern Georgia and northern Florida. All ponds and 28.2% of all samples (n = 635) were positive for Salmonella, with an overall geometric mean concentration (0.26 most probable number [MPN]/liter) that was relatively low compared to prior reports for rivers in this region. Salmonella peaks were seasonal; the levels correlated with increased temperature and rainfall (P < 0.05). The numbers and occurrence were significantly higher in water (0.32 MPN/liter and 37% of samples) than in sediment (0.22 MPN/liter and 17% of samples) but did not vary with depth. Representative isolates (n = 185) from different ponds, sample types, and seasons were examined for resistance to 15 different antibiotics; most strains were resistant to streptomycin (98.9%), while 20% were multidrug resistant (MDR) for 2 to 6 antibiotics. DiversiLab repetitive extragenic palindromic-element sequence-based PCR (rep-PCR) revealed genetic diversity and showed 43 genotypes among 191 isolates, as defined by >95% similarity. The genotypes did not partition by pond, season, or sample type. Genetic similarity to known serotypes indicated Hadar, Montevideo, and Newport as the most prevalent. All ponds achieved the current safety standards for generic Escherichia coli in agricultural water, and regression modeling showed that the E. coli level was a significant predictor for the probability of Salmonella occurrence. However, persistent populations of Salmonella were widely distributed in irrigation ponds, and the associated risks for produce contamination and subsequent human exposure are unknown, supporting continued surveillance of this pathogen in agricultural settings. PMID:25911476

  10. Distribution and Characterization of Salmonella enterica Isolates from Irrigation Ponds in the Southeastern United States

    PubMed Central

    Luo, Zhiyao; Gu, Ganyu; Ginn, Amber; Giurcanu, Mihai C.; Adams, Paige; Vellidis, George; van Bruggen, Ariena H. C.; Danyluk, Michelle D.

    2015-01-01

    Irrigation water has been implicated as a likely source of produce contamination by Salmonella enterica. Therefore, the distribution of S. enterica was surveyed monthly in irrigation ponds (n = 10) located within a prime agricultural region in southern Georgia and northern Florida. All ponds and 28.2% of all samples (n = 635) were positive for Salmonella, with an overall geometric mean concentration (0.26 most probable number [MPN]/liter) that was relatively low compared to prior reports for rivers in this region. Salmonella peaks were seasonal; the levels correlated with increased temperature and rainfall (P < 0.05). The numbers and occurrence were significantly higher in water (0.32 MPN/liter and 37% of samples) than in sediment (0.22 MPN/liter and 17% of samples) but did not vary with depth. Representative isolates (n = 185) from different ponds, sample types, and seasons were examined for resistance to 15 different antibiotics; most strains were resistant to streptomycin (98.9%), while 20% were multidrug resistant (MDR) for 2 to 6 antibiotics. DiversiLab repetitive extragenic palindromic-element sequence-based PCR (rep-PCR) revealed genetic diversity and showed 43 genotypes among 191 isolates, as defined by >95% similarity. The genotypes did not partition by pond, season, or sample type. Genetic similarity to known serotypes indicated Hadar, Montevideo, and Newport as the most prevalent. All ponds achieved the current safety standards for generic Escherichia coli in agricultural water, and regression modeling showed that the E. coli level was a significant predictor for the probability of Salmonella occurrence. However, persistent populations of Salmonella were widely distributed in irrigation ponds, and the associated risks for produce contamination and subsequent human exposure are unknown, supporting continued surveillance of this pathogen in agricultural settings. PMID:25911476

  11. Genomic and Phenotypic Analyses Reveal the Emergence of an Atypical Salmonella enterica Serovar Senftenberg Variant in China.

    PubMed

    Abd El Ghany, Moataz; Shi, Xiaolu; Li, Yinghui; Ansari, Hifzur R; Hill-Cawthorne, Grant A; Ho, Y S; Naeem, Raeece; Pickard, Derek; Klena, John D; Xu, Xuebing; Pain, Arnab; Hu, Qinghua

    2016-08-01

    Human infections with Salmonella enterica subspecies enterica serovar Senftenberg are often associated with exposure to poultry flocks, farm environments, or contaminated food. The recent emergence of multidrug-resistant isolates has raised public health concerns. In this study, comparative genomics and phenotypic analysis were used to characterize 14 Salmonella Senftenberg clinical isolates recovered from multiple outbreaks in Shenzhen and Shanghai, China, between 2002 and 2011. Single-nucleotide polymorphism analyses identified two phylogenetically distinct clades of S Senftenberg, designated SC1 and SC2, harboring variations in Salmonella pathogenicity island 1 (SPI-1) and SPI-2 and exhibiting distinct biochemical and phenotypic signatures. Although the two variants shared the same serotype, the SC2 isolates of sequence type 14 (ST14) harbored intact SPI-1 and -2 and hence were characterized by possessing efficient invasion capabilities. In contrast, the SC1 isolates had structural deletion patterns in both SPI-1 and -2 that correlated with an impaired capacity to invade cultured human cells and also the year of their isolation. These atypical SC1 isolates also lacked the capacity to produce hydrogen sulfide. These findings highlight the emergence of atypical Salmonella Senftenberg variants in China and provide genetic validation that variants lacking SPI-1 and regions of SPI-2, which leads to impaired invasion capacity, can still cause clinical disease. These data have identified an emerging public health concern and highlight the need to strengthen surveillance to detect the prevalence and transmission of nontyphoidal Salmonella species. PMID:27225410

  12. Another Bacillus sphaericus serotype harbouring strains very toxic to mosquito larvae: serotype H6.

    PubMed

    de Barjac, H; Thiery, I; Cosmao-Dumanoir, V; Frachon, E; Laurent, P; Charles, J F; Hamon, S; Ofori, J

    1988-01-01

    Ten isolates of Bacillus sphaericus from Ghana, very toxic to mosquito larvae, have been identified as belonging to serotype H6. These isolates can be represented by the head-group strain IAB59. They form crystals at the sporulation stage. Their larvicidal effect on Culex pipiens and Anopheles stephensi larvae is as high as that of the most toxic strains already known, e.g. 1593 and 2362 (serotype H5a,5b) and 2297 (serotype H25). Spore-crystal extracts of all these strains contain a 43-Kd polypeptide immunologically related to the 43-Kd polypeptide from strain 2362 described by other authors. PMID:3179062

  13. Complete Genome Sequence of Salmonella enterica subsp. enterica Serovar Typhi Isolate B/SF/13/03/195 Associated with a Typhoid Carrier in Pasir Mas, Kelantan, Malaysia

    PubMed Central

    Sim, Kee-Shin; Mohd Nor, Fauziah; Mat Hussin, Hani; Hamzah, Wan Mansor; Najimudin, Nazalan

    2015-01-01

    We report here the complete genome sequence of Salmonella enterica subsp. enterica serovar Typhi B/SF/13/03/195 obtained from a typhoid carrier, who is a food handler in Pasir Mas, Kelantan. PMID:26564035

  14. Novel serotype of bluetongue virus, western North America

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Bluetongue virus serotypes 10, 11, 13, and 17 are typically found throughout the United States (US), while serotype 2 was previously only detected in the southeastern US. In 2010, serotype 2 was identified in California for the first time and preliminary sequences analysis indicated that the virus ...

  15. Genome Sequence of Salmonella enterica subsp. enterica Serovar Typhi Isolate PM016/13 from Untreated Well Water Associated with a Typhoid Outbreak in Pasir Mas, Kelantan, Malaysia.

    PubMed

    Muhamad Harish, Salwani; Sim, Kee-Shin; Najimudin, Nazalan; Aziah, Ismail

    2015-01-01

    Salmonella enterica subsp. enterica serovar Typhi is a human-restricted pathogen that causes typhoid fever. Even though it is a human-restricted pathogen, the bacterium is also isolated from environments such as groundwater and pond water. Here, we describe the genome sequence of the Salmonella enterica subsp. enterica serovar Typhi PM016/13 which was isolated from well water during a typhoid outbreak in Kelantan, Malaysia, in 2013. PMID:26564032

  16. Genome Sequence of Salmonella enterica subsp. enterica Serovar Typhi Isolate PM016/13 from Untreated Well Water Associated with a Typhoid Outbreak in Pasir Mas, Kelantan, Malaysia

    PubMed Central

    Sim, Kee-Shin; Najimudin, Nazalan

    2015-01-01

    Salmonella enterica subsp. enterica serovar Typhi is a human-restricted pathogen that causes typhoid fever. Even though it is a human-restricted pathogen, the bacterium is also isolated from environments such as groundwater and pond water. Here, we describe the genome sequence of the Salmonella enterica subsp. enterica serovar Typhi PM016/13 which was isolated from well water during a typhoid outbreak in Kelantan, Malaysia, in 2013. PMID:26564032

  17. Draft Genome Sequences of Salmonella enterica subsp. enterica Serovar Berta ATCC 8392 and a Nalidixic Acid-Resistant Isolate of This Strain

    PubMed Central

    Cooper, Ashley; Koziol, Adam G.; Carrillo, Catherine D.

    2016-01-01

    Salmonella enterica subspecies enterica serovar Berta has been isolated in multiple animal species and has been implicated in human disease. Here, we report a 4.7-Mbp draft genome sequence of S. enterica serovar Berta (ATCC strain 8392) and a nalidixic acid-resistant isolate derived from this strain. PMID:27103707

  18. Identification of Haemophilus influenzae Serotypes by Standard Slide Agglutination Serotyping and PCR-Based Capsule Typing

    PubMed Central

    LaClaire, Leslye L.; Tondella, Maria Lucia C.; Beall, David S.; Noble, Corie A.; Raghunathan, Pratima L.; Rosenstein, Nancy E.; Popovic, Tanja

    2003-01-01

    To resolve discrepancies in slide agglutination serotyping (SAST) results from state health departments and the Centers for Disease Control and Prevention (CDC), we characterized 141 of 751 invasive Haemophilus influenzae isolates that were identified in the United States from January 1998 to December 1999 through an active, laboratory-based, surveillance program coordinated by the CDC. We found discrepancies between the results of SAST performed at state health departments and those of PCR capsule typing performed at the CDC for 56 (40%) of the isolates characterized: 54 isolates that were identified as a particular serotype by SAST were shown to be unencapsulated by PCR, and two isolates that were reported as serotypes b and f were found to be serotypes f and e, respectively, by PCR. The laboratory error most likely to affect the perceived efficacy of the conjugate H. influenzae type b (Hib) vaccine was the misidentification of isolates as serotype b: of 40 isolates identified as serotype b by SAST, 27 (68%) did not contain the correlating capsule type genes. The frequency of errors fell substantially when standardized reagents and routine quality control of SAST were used during a study involving three laboratories. An overall 94% agreement between SAST and PCR results showed that slide agglutination could be a valid and reliable method for serotyping H. influenzae if the test was performed correctly, in accordance with standardized and recommended procedures. An ongoing prospective analysis of all H. influenzae surveillance isolates associated with invasive disease in children less than 5 years old will provide more accurate national figures for the burden of invasive disease caused by Hib and other H. influenzae serotypes. PMID:12517878

  19. Novel Bluetongue Virus Serotype from Kuwait

    PubMed Central

    Maan, Sushila; Maan, Narender S.; Nomikou, Kyriaki; Batten, Carrie; Antony, Frank; Belaganahalli, Manjunatha N.; Samy, Attia Mohamed; Abdel Reda, Ammar; Al-Rashid, Sana Ahmed; El Batel, Maha; Oura, Chris A.L.

    2011-01-01

    Sheep and goats sampled in Kuwait during February 2010 were seropositive for bluetongue virus (BTV). BTV isolate KUW2010/02, from 1 of only 2 sheep that also tested positive for BTV by real-time reverse transcription–PCR, caused mild clinical signs in sheep. Nucleotide sequencing identified KUW2010/02 as a novel BTV serotype. PMID:21529403

  20. Novel bluetongue virus serotype from Kuwait.

    PubMed

    Maan, Sushila; Maan, Narender S; Nomikou, Kyriaki; Batten, Carrie; Antony, Frank; Belaganahalli, Manjunatha N; Samy, Attia Mohamed; Reda, Ammar Abdel; Al-Rashid, Sana Ahmed; El Batel, Maha; Oura, Chris A L; Mertens, Peter P C

    2011-05-01

    Sheep and goats sampled in Kuwait during February 2010 were seropositive for bluetongue virus (BTV). BTV isolate KUW2010/02, from 1 of only 2 sheep that also tested positive for BTV by real-time reverse transcription-PCR, caused mild clinical signs in sheep. Nucleotide sequencing identified KUW2010/02 as a novel BTV serotype. PMID:21529403

  1. Distribution of serotypes of human rotavirus in different populations.

    PubMed Central

    Woods, P A; Gentsch, J; Gouvea, V; Mata, L; Santosham, M; Bai, Z S; Urasawa, S; Glass, R I

    1992-01-01

    Serotyping is a useful tool to study the epidemiologic characteristics of rotaviruses in large populations and to assess the need for a vaccine to protect against all strains. By using an enzyme immunoassay with serotype-specific monoclonal antibodies to the four most common rotavirus serotypes, we analyzed 1,183 rotavirus-positive specimens from 16 stool collections in eight countries on four continents that were obtained from 1978 to 1989. Of the 926 strains (78%) that could be serotyped, 48% were serotype 1, 8% were serotype 2, 15% were serotype 3, and 7% were serotype 4. Twenty-two percent had insufficient numbers of double-shelled virus particles to react with the monoclonal antibody of the VP4 rotavirus protein and therefore could not be serotyped. Our results indicate that vaccines being developed must provide the greatest coverage against serotype 1 and that the serotype distribution cannot be predicted currently by the geographic area or prevalence in the preceding year. PMID:1315333

  2. Selective Enrichment Media Bias the Types of Salmonella enterica Strains Isolated from Mixed Strain Cultures and Complex Enrichment Broths

    PubMed Central

    Gorski, Lisa

    2012-01-01

    For foodborne outbreak investigations it can be difficult to isolate the relevant strain from food and/or environmental sources. If the sample is contaminated by more than one strain of the pathogen the relevant strain might be missed. In this study mixed cultures of Salmonella enterica were grown in one set of standard enrichment media to see if culture bias patterns emerged. Nineteen strains representing four serogroups and ten serotypes were compared in four-strain mixtures in Salmonella-only and in cattle fecal culture enrichment backgrounds using Salmonella enrichment media. One or more strain(s) emerged as dominant in each mixture. No serotype was most fit, but strains of serogroups C2 and E were more likely to dominate enrichment culture mixtures than strains of serogroups B or C1. Different versions of Rappaport-Vassiliadis (RV) medium gave different patterns of strain dominance in both Salmonella-only and fecal enrichment culture backgrounds. The fittest strains belonged to serogroups C1, C2, and E, and included strains of S. Infantis, S. Thompson S. Newport, S. 6,8:d:-, and S. Give. Strains of serogroup B, which included serotypes often seen in outbreaks such as S. Typhimurium, S. Saintpaul, and S. Schwarzengrund were less likely to emerge as dominant strains in the mixtures when using standard RV as part of the enrichment. Using a more nutrient-rich version of RV as part of the protocol led to a different pattern of strains emerging, however some were still present in very low numbers in the resulting population. These results indicate that outbreak investigations of food and/or other environmental samples should include multiple enrichment protocols to ensure isolation of target strains of Salmonella. PMID:22496847

  3. Interaction of Salmonella enterica with Fresh Produce Leaves

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Attachment and colonization of Salmonella enterica serovars to fresh produce leaves was investigated. Biofilm assay and attachment of Salmonella serovars to intact and cut leaves were determined. Salmonella Tennessee and Salmonella Thompson produced stronger biofilms compared to Salmonella Newpor...

  4. High-throughput Assay to Phenotype Salmonella enterica Typhimurium Association, Invasion, and Replication in Macrophages

    PubMed Central

    Wu, Jing; Pugh, Roberta; Laughlin, Richard C.; Andrews-Polymenis, Helene; McClelland, Michael; Bäumler, Andreas J.; Adams, L. Garry

    2014-01-01

    Salmonella species are zoonotic pathogens and leading causes of food borne illnesses in humans and livestock1. Understanding the mechanisms underlying Salmonella-host interactions are important to elucidate the molecular pathogenesis of Salmonella infection. The Gentamicin protection assay to phenotype Salmonella association, invasion and replication in phagocytic cells was adapted to allow high-throughput screening to define the roles of deletion mutants of Salmonella enterica serotype Typhimurium in host interactions using RAW 264.7 murine macrophages. Under this protocol, the variance in measurements is significantly reduced compared to the standard protocol, because wild-type and multiple mutant strains can be tested in the same culture dish and at the same time. The use of multichannel pipettes increases the throughput and enhances precision. Furthermore, concerns related to using less host cells per well in 96-well culture dish were addressed. Here, the protocol of the modified in vitro Salmonella invasion assay using phagocytic cells was successfully employed to phenotype 38 individual Salmonella deletion mutants for association, invasion and intracellular replication. The in vitro phenotypes are presented, some of which were subsequently confirmed to have in vivo phenotypes in an animal model. Thus, the modified, standardized assay to phenotype Salmonella association, invasion and replication in macrophages with high-throughput capacity could be utilized more broadly to study bacterial-host interactions. PMID:25146526

  5. Survival of Salmonella enterica in Dried Turkey Manure and Persistence on Spinach Leaves.

    PubMed

    Oni, Ruth A; Sharma, Manan; Buchanan, Robert L

    2015-10-01

    Concerns about the microbiological safety of fresh produce have attracted attention in the past three decades due to multiple foodborne outbreaks. Animal manure contaminated with enteric pathogens has been identified as an important preharvest pathogen source. This study investigated the survival of Salmonella enterica in dust particles of dehydrated turkey manure and how association with manure dust may enhance the survival of salmonellae on leafy greens in the field. The survival of a cocktail of multiple Salmonella serotypes in the dried fecal material of various particle sizes (125 to 500 μm) was examined at varying moisture contents (5, 10, and 15%). Survival times of the pathogen were inversely related to moisture content and particle size of manure dust, with viable Salmonella still detectable for up to 291 days in the smallest particle size (125 μm) with 5% moisture. Association with manure dust particles increased the survival of Salmonella when subjected to UV light both under laboratory conditions and on the surface of spinach leaves in a greenhouse setting. The results of this study suggest that aerosolized manure particles could be a potential vehicle for Salmonella dispersal to leafy greens if the microorganism is present in the dry manure. PMID:26408127

  6. Identification of newly recognized serotype 1c as the most prevalent Shigella flexneri serotype in northern rural Vietnam

    PubMed Central

    STAGG, R. M.; CAM, P. D.; VERMA, N. K.

    2008-01-01

    SUMMARY We investigated the identity of 37 Shigella flexneri strains that had previously been isolated from northern rural Vietnam (Son Tay Province) and described as untypable. Twenty-four isolates reacted with MASF 1c, a monoclonal antibody specific for S. flexneri serotype 1c. A further ten untypable isolates were found to be rough mutants (no longer expressing O-antigen) that were derived from serotype 1c strains. Pulsed-field gel electrophoresis demonstrated that these strains consisted of many different clones, indicating serotype 1c was well established in this region in the late 1990s. Serotype 1c was the most prevalent S. flexneri serotype isolated in the Son Tay Province, accounting for about 40% of S. flexneri isolates. Subsequent isolation of S. flexneri serotype 1c in this region and elsewhere in Vietnam confirmed that serotype 1c is of genuine importance in Vietnam. PMID:17922932

  7. Emerging resistant serotypes of invasive Streptococcus pneumoniae

    PubMed Central

    Elshafie, Sittana; Taj-Aldeen, Saad J

    2016-01-01

    Background Streptococcus pneumoniae is the leading cause of meningitis and sepsis. The aim of the study was to analyze the distribution, vaccine serotype coverage, and antibiotic resistance of S. pneumoniae serotypes isolated from patients with invasive diseases, after the introduction of pneumococcal 7-valent conjugated vaccine (PCV-7). Methods A total of 134 isolates were collected from blood and cerebrospinal fluid specimens at Hamad Hospital during the period from 2005 to 2009. Isolate serotyping was done using the Quellung reaction. The prevaccination period was considered before 2005. Results The most common serotypes for all age groups were 3 (12.70%), 14 (11.90%), 1 (11.90%), 19A (9.00%), 9V (5.20%), 23F (5.20%), and 19F (4.50%). Coverage rates for infant <2 years for PCV-7, the 10-valent conjugated vaccine (PCV-10), and the 13-valent conjugated vaccine (PCV-13) were 34.78%, 52.17%, and 78.26%, respectively. Coverage rates of these vaccines were 50%, 67.86%, and 75% for the 2–5 years age group; 27.12%, 40.68%, and 64.41% for the age group 6–64 years; and 25%, 33.33%, and 66.67% for the ≥65 years age group, respectively. The percentage of nonsusceptible isolates to penicillin, cefotaxime, and erythromycin were 43.86%, 16.66%, and 22.81%, respectively. Thirty-seven isolates (32.46%) were multidrug resistant (MDR) and belonged to serotypes 14, 19A, 19F, 23F, 1, 9V, 12F, 4, 6B, 3, and 15A. Compared to previous results before the introduction of PCV-7, there was a significant reduction in penicillin-nonsusceptable S. pneumoniae from 66.67% to 43.86%, and a slight insignificant reduction in erythromycin nonsusceptible strains from 27.60% to 22.8%, while there was a significant increase in cefotaxime nonsusceptible strains from 3.55% to 16.66%. Conclusion Invasive pneumococcal strains and the emergence of MDR serotypes is a global burden that must be addressed through multiple strategies, including vaccination, antibiotic stewardship, and continuous

  8. Salmonella Serotype Determination Utilizing High-Throughput Genome Sequencing Data

    PubMed Central

    Zhang, Shaokang; Yin, Yanlong; Jones, Marcus B.; Zhang, Zhenzhen; Deatherage Kaiser, Brooke L.; Dinsmore, Blake A.; Fitzgerald, Collette; Fields, Patricia I.

    2015-01-01

    Serotyping forms the basis of national and international surveillance networks for Salmonella, one of the most prevalent foodborne pathogens worldwide (1–3). Public health microbiology is currently being transformed by whole-genome sequencing (WGS), which opens the door to serotype determination using WGS data. SeqSero (www.denglab.info/SeqSero) is a novel Web-based tool for determining Salmonella serotypes using high-throughput genome sequencing data. SeqSero is based on curated databases of Salmonella serotype determinants (rfb gene cluster, fliC and fljB alleles) and is predicted to determine serotype rapidly and accurately for nearly the full spectrum of Salmonella serotypes (more than 2,300 serotypes), from both raw sequencing reads and genome assemblies. The performance of SeqSero was evaluated by testing (i) raw reads from genomes of 308 Salmonella isolates of known serotype; (ii) raw reads from genomes of 3,306 Salmonella isolates sequenced and made publicly available by GenomeTrakr, a U.S. national monitoring network operated by the Food and Drug Administration; and (iii) 354 other publicly available draft or complete Salmonella genomes. We also demonstrated Salmonella serotype determination from raw sequencing reads of fecal metagenomes from mice orally infected with this pathogen. SeqSero can help to maintain the well-established utility of Salmonella serotyping when integrated into a platform of WGS-based pathogen subtyping and characterization. PMID:25762776

  9. Evaluation of Molecular Methods for Serotyping Shigella flexneri.

    PubMed

    Gentle, Amy; Ashton, Philip M; Dallman, Timothy J; Jenkins, Claire

    2016-06-01

    Shigella flexneri can be phenotypically serotyped using antisera raised to type-specific somatic antigens and group factor antigens and genotypically serotyped using PCR targeting O-antigen synthesis or modification genes. The aim of this study was to evaluate a real-time PCR for serotyping S. flexneri and to use whole-genome sequencing (WGS) to investigate the phenotypic and genotypic serotype identifications. Of the 244 cultures tested retrospectively, 226 (92.6%) had concordant results between phenotypic serotyping and PCR. Seventy of the 244 isolates (including 15 of the 18 isolates where a serotype-PCR mismatch was identified) were whole-genome sequenced, and the serotype was derived from the genome. Discrepant results between the phenotypic and genotypic tests were attributed to insertions/deletions or point mutations identified in O-antigen synthesis or modification genes, rendering them dysfunctional; inconclusive serotyping results due to nonspecific cross-reactions; or novel genotypes. Phylogenetic analysis of the WGS data indicated that the serotype, regardless of whether it was phenotypically or genotypically determined, was a weak predictor of phylogenetic relationships between strains of S. flexneri WGS data provided both genome-derived serotyping, thus supporting backward compatibility with historical data and facilitating data exchange in the community, and more robust and discriminatory typing at the single-nucleotide-polymorphism level. PMID:26984974

  10. Epidemiology of Bluetongue Virus Serotype 8, Germany

    PubMed Central

    Gethmann, Jörn M.; Staubach, Christoph; Mettenleiter, Thomas C.; Beer, Martin; Hoffmann, Bernd

    2009-01-01

    In Germany, bluetongue disease had not been reported before 2006. During August 2006–August 2008, >24,000 bluetongue virus serotype 8 infections were reported, most (20,635) in 2007. In 2006 and 2007, respectively, case-fatality rates were 6.4% and 13.1% for cattle and 37.5% and 41.5% for sheep. Vaccination in 2008 decreased cases. PMID:19239757

  11. Multistate outbreak of Salmonella serotype Bovismorbificans infections associated with hummus and tahini--United States, 2011.

    PubMed

    2012-11-23

    On September 27, 2011, three clinical isolates of Salmonella enterica serotype Bovismorbificans with indistinguishable pulsed-field gel electrophoresis (PFGE) patterns were identified by the District of Columbia Public Health Laboratory (PHL). Human infection with S. Bovismorbificans is rare in the United States. Through query of PulseNet, the national molecular subtyping network for foodborne disease surveillance, six additional cases with indistinguishable PFGE patterns were identified in three states (Maryland, Michigan, and Virginia) during the prior 60 days. All nine patients had eaten at restaurants in the District of Columbia (DC) or northern Virginia <2 weeks before illness onset. This report summarizes the investigation led by the DC Department of Health (DOH), in which 23 cases of S. Bovismorbificans infections were identified among persons from seven states and DC, with illness onset during August 19-November 21, 2011. On May 30, 2012, traceback indicated that contaminated tahini (sesame seed paste) used in hummus prepared at a Mediterranean-style restaurant in DC was a plausible source of Salmonella infections. DOH restricted the sale of hummus and prohibited the use of hummus ingredients in other food items at implicated restaurants to prevent further illness. This investigation also illustrates challenges associated with ingredient-driven outbreaks and the value of PulseNet for identifying clusters of cases that are geographically dispersed. PMID:23169315

  12. Diversity of Genome Structure in Salmonella enterica Serovar Typhi Populations†

    PubMed Central

    Kothapalli, Sushma; Nair, Satheesh; Alokam, Suneetha; Pang, Tikki; Khakhria, Rasik; Woodward, David; Johnson, Wendy; Stocker, Bruce A. D.; Sanderson, Kenneth E.; Liu, Shu-Lin

    2005-01-01

    The genomes of most strains of Salmonella and Escherichia coli are highly conserved. In contrast, all 136 wild-type strains of Salmonella enterica serovar Typhi analyzed by partial digestion with I-CeuI (an endonuclease which cuts within the rrn operons) and pulsed-field gel electrophoresis and by PCR have rearrangements due to homologous recombination between the rrn operons leading to inversions and translocations. Recombination between rrn operons in culture is known to be equally frequent in S. enterica serovar Typhi and S. enterica serovar Typhimurium; thus, the recombinants in S. enterica serovar Typhi, but not those in S. enterica serovar Typhimurium, are able to survive in nature. However, even in S. enterica serovar Typhi the need for genome balance and the need for gene dosage impose limits on rearrangements. Of 100 strains of genome types 1 to 6, 72 were only 25.5 kb off genome balance (the relative lengths of the replichores during bidirectional replication from oriC to the termination of replication [Ter]), while 28 strains were less balanced (41 kb off balance), indicating that the survival of the best-balanced strains was greater. In addition, the need for appropriate gene dosage apparently selected against rearrangements which moved genes from their accustomed distance from oriC. Although rearrangements involving the seven rrn operons are very common in S. enterica serovar Typhi, other duplicated regions, such as the 25 IS200 elements, are very rarely involved in rearrangements. Large deletions and insertions in the genome are uncommon, except for deletions of Salmonella pathogenicity island 7 (usually 134 kb) from fragment I-CeuI-G and 40-kb insertions, possibly a prophage, in fragment I-CeuI-E. The phage types were determined, and the origins of the phage types appeared to be independent of the origins of the genome types. PMID:15805510

  13. Salmonella enterica Diversity in Central Californian Coastal Waterways

    PubMed Central

    Walters, Sarah P.; González-Escalona, Narjol; Son, Insook; Melka, David C.; Sassoubre, Lauren M.

    2013-01-01

    Salmonella enterica is one of the most important bacterial enteric pathogens worldwide. However, little is known about its distribution and diversity in the environment. The present study explored the diversity of 104 strains of Salmonella enterica isolated over 2 years from 12 coastal waterways in central California. Pulsed-field gel electrophoresis (PFGE) and multilocus sequence typing were used to probe species diversity. Seventy-four PFGE patterns and 38 sequence types (STs) were found, including 18 newly described STs. Nineteen of 25 PFGE patterns were indistinguishable from those of clinical isolates in PulseNet. The most common ST was consistent with S. enterica serovar Typhimurium, and other frequently detected STs were associated with the serovars Heidelberg and Enteritidis; all of these serovars are important etiologies of salmonellosis. An investigation into S. enterica biogeography was conducted at the level of ST and subspecies. At the ST and subspecies level, we found a taxon-time relationship but no taxon-area or taxon-environmental distance relationships. STs collected during wet versus dry conditions tended to be more similar; however, STs collected from waterways adjacent to watersheds with similar land covers did not tend to be similar. The results suggest that the lack of dispersal limitation may be an important factor affecting the diversity of S. enterica in the region. PMID:23624479

  14. Genomic comparison of Salmonella enterica serotypes commonly associated with cattle and beef – Analysis of variation in virulence potential

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Category: Post-Harvest Abstract Published: Unpublished to date Objective: To create a Salmonella genomic DNA sequence resource for the purpose of characterizing differences in gene content and identification of DNA markers for rapid detection of virulent Salmonella strains. Further, to character...

  15. Cellular and Cytokine Responses to Salmonella enterica Serotype Typhi Proteins in Patients with Typhoid Fever in Bangladesh

    PubMed Central

    Sayeed, Abu; Khanam, Farhana; Leung, Daniel T.; Rahman Bhuiyan, Taufiqur; Sheikh, Alaullah; Salma, Umme; LaRocque, Regina C.; Harris, Jason B.; Pacek, Marcin; Calderwood, Stephen B.; LaBaer, Joshua; Charles, Richelle C.

    2014-01-01

    We assessed interferon-gamma (IFN-γ) responses via enzyme-linked immunosorbent spot (ELISPOT) to a number of S. Typhi antigens in samples from humans with S. Typhi bacteremia and typhoid fever in Bangladesh. Compared with responses in healthy endemic zone controls, there were significantly increased IFN-γ responses at the time of clinical presentation (acute phase) and at convalescence 14–28 days later. The majority (80–90%) of IFN-γ expressing T cells were CD4+. We observed a significant increase in interleukin-17 (IL-17) positive CD4 + T cells at convalescent versus acute stage of infection using an intracellular cytokine staining assay. We also found that stimulated peripheral blood mononuclear cells (PBMCs) produced significantly increased levels of a number of cytokines at the convalescent versus acute phase of infection, including IFN-γ, MIP-1β, sCD40L, TNF-β, IL-13, and IL-9. These results suggest that S. Typhi antigens induce a predominantly Th1 response, but that elevations in other cytokines may be modulatory. PMID:24615129

  16. Rapid Multiplex PCR and Real-Time TaqMan PCR Assays for Detection of Salmonella enterica and the Highly Virulent Serovars Choleraesuis and Paratyphi C▿ †

    PubMed Central

    Woods, David F.; Reen, F. Jerry; Gilroy, Deirdre; Buckley, Jim; Frye, Jonathan G.; Boyd, E. Fidelma

    2008-01-01

    Salmonella enterica is a human pathogen with over 2,500 serovars characterized. S. enterica serovars Choleraesuis and Paratyphi C are two globally distributed serovars. We have developed a rapid molecular-typing method to detect serovars Choleraesuis and Paratyphi C in food samples by using a comparative-genomics approach to identify regions unique to each serovar from the sequenced genomes. A Salmonella-specific primer pair based on oriC was designed as an internal control to establish accuracy, sensitivity, and reproducibility. Serovar-specific primer sets based on regions of difference between serovars Choleraesuis and Paratyphi C were designed for real-time PCR assays. Three primer sets were used to screen a collection of over 100 Salmonella strains, and both serovars Choleraesuis and Paratyphi C gave unique amplification patterns. To develop the technique for practical use, its sensitivity for detection of Salmonella spp. in a food matrix was determined by spiking experiments. The technique was also adapted for a real-time PCR rapid-detection assay for both serovars Choleraesuis and Paratyphi C that complements the current procedures for Salmonella sp. isolation and serotyping. PMID:18923008

  17. Elucidation of Antimicrobial Susceptibility Profiles and Genotyping of Salmonella enterica Isolates from Clinical Cases of Salmonellosis in New Mexico in 2008

    PubMed Central

    Smith, Kenneth P.; George, Jeffy; Cadle, Kathleen M.; Kumar, Sanath; Aragon, Steven J.; Hernandez, Ricardo L.; Jones, Suzanna E.; Floyd, Jody L.; Varela, Manuel F.

    2010-01-01

    In this study, we investigated the antimicrobial susceptibility profiles and the distribution of some well known genetic determinants of virulence in clinical isolates of Salmonella enterica from New Mexico. The minimum inhibitory concentrations (MICs) for various antimicrobials were determined by using the E-test strip method according to CLSI guidelines. Virulence genotyping was performed by polymerase chain reaction (PCR) using primers specific for known virulence genes of Salmonella enterica. Of 15 isolates belonging to 11 different serovars analyzed, one isolate of Salmonella Typhimurium was resistant to multiple drugs namely ampicillin, amoxicillin / clavulanic acid, chloramphenicol and tetracycline, that also harbored class 1 intergron, blaTEM encoding genes for β-lactamase, chloramphenicol acetyl transferase (cat1), plus floR, tet(C) and tet(G). This strain was phage typed as DT104. PCR analysis revealed the presence of invA, hilA, stn, agfA and spvR virulence genes in all the isolates tested. The plasmid-borne pefA gene was absent in 11 isolates, while 5 isolates lacked sopE. One isolate belonging to serogroup E4 (Salmonella Sombre) was devoid of multiple virulence genes pefA, iroB, shdA and sopE. These results demonstrate that clinical Salmonella serotypes from New Mexico used here are predominantly sensitive to multiple antimicrobial agents, but vary in their virulence genotypes. Information on antimicrobial sensitivity and virulence genotypes will help in understanding the evolution and spread of epidemic strains of Salmonella enterica in the region of study. PMID:20514366

  18. Salmonella enterica: Survival, Colonization, and Virulence Differences among Serovars

    PubMed Central

    Andino, A.; Hanning, I.

    2015-01-01

    Data indicate that prevalence of specific serovars of Salmonella enterica in human foodborne illness is not correlated with their prevalence in feed. Given that feed is a suboptimal environment for S. enterica, it appears that survival in poultry feed may be an independent factor unrelated to virulence of specific serovars of Salmonella. Additionally, S. enterica serovars appear to have different host specificity and the ability to cause disease in those hosts is also serovar dependent. These differences among the serovars may be related to gene presence or absence and expression levels of those genes. With a better understanding of serovar specificity, mitigation methods can be implemented to control Salmonella at preharvest and postharvest levels. PMID:25664339

  19. Growth kinetics of Salmonella enterica in Hajna tetrathionate broth, Rappaport broth and modified semisolid Rappaport agar

    PubMed Central

    FUJIHARA, Masatoshi; TABUCHI, Hiroyuki; UEGAKI, Kaho

    2015-01-01

    To determine the appropriate method for isolating Salmonella enterica, we compared the growth of S. enterica serovars using three selective enrichment media. S. enterica was more successfully isolated from artificially contaminated fecal samples after enrichment in Hajna tetrathionate broth or modified semisolid Rappaport agar than in Rappaport broth. Since most bacteria (other than motile S. enterica) do not migrate on modified semisolid Rappaport agar, the growth characteristics of S. enterica can be interpreted easily and quickly. Two S. enterica isolates did not migrate on modified semisolid Rappaport agar, but did grow in Hajna tetrathionate broth, which suggests that the combined use of these selective enrichment media is appropriate for isolating S. enterica. PMID:26498402

  20. Internal Colonization of Salmonella enterica Serovar Typhimurium in Tomato Plants

    PubMed Central

    Gu, Ganyu; Hu, Jiahuai; Cevallos-Cevallos, Juan M.; Richardson, Susanna M.; Bartz, Jerry A.; van Bruggen, Ariena H. C.

    2011-01-01

    Several Salmonella enterica outbreaks have been traced back to contaminated tomatoes. In this study, the internalization of S. enterica Typhimurium via tomato leaves was investigated as affected by surfactants and bacterial rdar morphotype, which was reported to be important for the environmental persistence and attachment of Salmonella to plants. Surfactants, especially Silwet L-77, promoted ingress and survival of S. enterica Typhimurium in tomato leaves. In each of two experiments, 84 tomato plants were inoculated two to four times before fruiting with GFP-labeled S. enterica Typhimurium strain MAE110 (with rdar morphotype) or MAE119 (without rdar). For each inoculation, single leaflets were dipped in 109 CFU/ml Salmonella suspension with Silwet L-77. Inoculated and adjacent leaflets were tested for Salmonella survival for 3 weeks after each inoculation. The surface and pulp of ripe fruits produced on these plants were also examined for Salmonella. Populations of both Salmonella strains in inoculated leaflets decreased during 2 weeks after inoculation but remained unchanged (at about 104 CFU/g) in week 3. Populations of MAE110 were significantly higher (P<0.05) than those of MAE119 from day 3 after inoculation. In the first year, nine fruits collected from one of the 42 MAE119 inoculated plants were positive for S. enterica Typhimurium. In the second year, Salmonella was detected in adjacent non-inoculated leaves of eight tomato plants (five inoculated with strain MAE110). The pulp of 12 fruits from two plants inoculated with MAE110 was Salmonella positive (about 106 CFU/g). Internalization was confirmed by fluorescence and confocal laser microscopy. For the first time, convincing evidence is presented that S. enterica can move inside tomato plants grown in natural field soil and colonize fruits at high levels without inducing any symptoms, except for a slight reduction in plant growth. PMID:22096553

  1. Colonization and internalization of Salmonella enterica in tomato plants.

    PubMed

    Zheng, Jie; Allard, Sarah; Reynolds, Sara; Millner, Patricia; Arce, Gabriela; Blodgett, Robert J; Brown, Eric W

    2013-04-01

    The consumption of fresh tomatoes has been linked to numerous food-borne outbreaks involving various serovars of Salmonella enterica. Recent advances in our understanding of plant-microbe interactions have shown that human enteric pathogenic bacteria, including S. enterica, are adapted to survive in the plant environment. In this study, tomato plants (Solanum lycopersicum cv. Micro-Tom) grown in sandy loam soil from Virginia's eastern shore (VES) were inoculated with S. enterica serovars to evaluate plausible internalization routes and to determine if there is any niche fitness for certain serovars. Both infested soil and contaminated blossoms can lead to low internal levels of fruit contamination with Salmonella. Salmonella serovars demonstrated a great ability to survive in environments under tomato cultivation, not only in soil but also on different parts of the tomato plant. Of the five serovars investigated, Salmonella enterica serovars Newport and Javiana were dominant in sandy loam soil, while Salmonella enterica serovars Montevideo and Newport were more prevalent on leaves and blossoms. It was also observed that Salmonella enterica serovar Typhimurium had a poor rate of survival in all the plant parts examined here, suggesting that postharvest contamination routes are more likely in S. Typhimurium contamination of tomato fruit. Conversely, S. Newport was the most prevalent serovar recovered in both the tomato rhizosphere and phyllosphere. Plants that were recently transplanted (within 3 days) had an increase in observable internalized bacteria, suggesting that plants were more susceptible to internalization right after transplant. These findings suggest that the particular Salmonella serovar and the growth stage of the plant were important factors for internalization through the root system. PMID:23377940

  2. Colonization and Internalization of Salmonella enterica in Tomato Plants

    PubMed Central

    Allard, Sarah; Reynolds, Sara; Millner, Patricia; Arce, Gabriela; Blodgett, Robert J.; Brown, Eric W.

    2013-01-01

    The consumption of fresh tomatoes has been linked to numerous food-borne outbreaks involving various serovars of Salmonella enterica. Recent advances in our understanding of plant-microbe interactions have shown that human enteric pathogenic bacteria, including S. enterica, are adapted to survive in the plant environment. In this study, tomato plants (Solanum lycopersicum cv. Micro-Tom) grown in sandy loam soil from Virginia's eastern shore (VES) were inoculated with S. enterica serovars to evaluate plausible internalization routes and to determine if there is any niche fitness for certain serovars. Both infested soil and contaminated blossoms can lead to low internal levels of fruit contamination with Salmonella. Salmonella serovars demonstrated a great ability to survive in environments under tomato cultivation, not only in soil but also on different parts of the tomato plant. Of the five serovars investigated, Salmonella enterica serovars Newport and Javiana were dominant in sandy loam soil, while Salmonella enterica serovars Montevideo and Newport were more prevalent on leaves and blossoms. It was also observed that Salmonella enterica serovar Typhimurium had a poor rate of survival in all the plant parts examined here, suggesting that postharvest contamination routes are more likely in S. Typhimurium contamination of tomato fruit. Conversely, S. Newport was the most prevalent serovar recovered in both the tomato rhizosphere and phyllosphere. Plants that were recently transplanted (within 3 days) had an increase in observable internalized bacteria, suggesting that plants were more susceptible to internalization right after transplant. These findings suggest that the particular Salmonella serovar and the growth stage of the plant were important factors for internalization through the root system. PMID:23377940

  3. Multi-Serotype Pneumococcal Nasopharyngeal Carriage Prevalence in Vaccine Naïve Nepalese Children, Assessed Using Molecular Serotyping

    PubMed Central

    Kandasamy, Rama; Gurung, Meeru; Thapa, Anushil; Ndimah, Susan; Adhikari, Neelam; Murdoch, David R.; Kelly, Dominic F.; Waldron, Denise E.; Gould, Katherine A.; Thorson, Stephen; Shrestha, Shrijana; Hinds, Jason; Pollard, Andrew J.

    2015-01-01

    Invasive pneumococcal disease is one of the major causes of death in young children in resource poor countries. Nasopharyngeal carriage studies provide insight into the local prevalence of circulating pneumococcal serotypes. There are very few data on the concurrent carriage of multiple pneumococcal serotypes. This study aimed to identify the prevalence and serotype distribution of pneumococci carried in the nasopharynx of young healthy Nepalese children prior to the introduction of a pneumococcal conjugate vaccine using a microarray-based molecular serotyping method capable of detecting multi-serotype carriage. We conducted a cross-sectional study of healthy children aged 6 weeks to 24 months from the Kathmandu Valley, Nepal between May and October 2012. Nasopharyngeal swabs were frozen and subsequently plated on selective culture media. DNA extracts of plate sweeps of pneumococcal colonies from these cultures were analysed using a molecular serotyping microarray capable of detecting relative abundance of multiple pneumococcal serotypes. 600 children were enrolled into the study: 199 aged 6 weeks to <6 months, 202 aged 6 months to < 12 months, and 199 aged 12 month to 24 months. Typeable pneumococci were identified in 297/600 (49·5%) of samples with more than one serotype being found in 67/297 (20·2%) of these samples. The serotypes covered by the thirteen-valent pneumococcal conjugate vaccine were identified in 44·4% of samples containing typeable pneumococci. Application of a molecular serotyping approach to identification of multiple pneumococcal carriage demonstrates a substantial prevalence of co-colonisation. Continued surveillance utilising this approach following the introduction of routine use of pneumococcal conjugate vaccinates in infants will provide a more accurate understanding of vaccine efficacy against carriage and a better understanding of the dynamics of subsequent serotype and genotype replacement. PMID:25643355

  4. Molecular Serotyping of Escherichia coli O111:H8

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Accurate Escherichia coli serotyping is critical for pathogen diagnosis and surveillance of non-O157 shiga-toxigenic strains, however, few laboratories have this capacity. The molecular serotyping protocol described in this paper targets the somatic and flagellar antigens of E. coli O111:H8 used in...

  5. Rotavirus serotype G5 associated with diarrhea in Brazilian children.

    PubMed

    Gouvea, V; de Castro, L; Timenetsky, M C; Greenberg, H; Santos, N

    1994-05-01

    Rotavirus serotype G5 in fecal specimens of 38 Brazilian children with diarrhea was identified by PCR and enzyme immunoassays. The strains exhibited long RNA electropherotypes and either subgroup II or nonsubgroup I-nonsubgroup II specificities. Serotype G5 has been found in piglets and horses but not yet in humans. PMID:8051281

  6. Probability of identifying different salmonella serotypes in poultry samples

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Recent work has called attention to the unequal competitive abilities of different Salmonella serotypes in standard broth culture and plating media. Such serotypes include Enteritidis and Typhimurium that are specifically targeted in some regulatory and certification programs because they cause a l...

  7. Plant Pathogen-Induced Water-Soaking Promotes Salmonella enterica Growth on Tomato Leaves

    PubMed Central

    Potnis, Neha; Colee, James; Jones, Jeffrey B.

    2015-01-01

    Plant pathogen infection is a critical factor for the persistence of Salmonella enterica on plants. We investigated the mechanisms responsible for the persistence of S. enterica on diseased tomato plants by using four diverse bacterial spot Xanthomonas species that differ in disease severities. Xanthomonas euvesicatoria and X. gardneri infection fostered S. enterica growth, while X. perforans infection did not induce growth but supported the persistence of S. enterica. X. vesicatoria-infected leaves harbored S. enterica populations similar to those on healthy leaves. Growth of S. enterica was associated with extensive water-soaking and necrosis in X. euvesicatoria- and X. gardneri-infected plants. The contribution of water-soaking to the growth of S. enterica was corroborated by an increased growth of populations on water-saturated leaves in the absence of a plant pathogen. S. enterica aggregates were observed with bacterial spot lesions caused by either X. euvesicatoria or X. vesicatoria; however, more S. enterica aggregates formed on X. euvesicatoria-infected leaves as a result of larger lesion sizes per leaf area and extensive water-soaking. Sparsely distributed lesions caused by X. vesicatoria infection do not support the overall growth of S. enterica or aggregates in areas without lesions or water-soaking; S. enterica was observed as single cells and not aggregates. Thus, pathogen-induced water-soaking and necrosis allow S. enterica to replicate and proliferate on tomato leaves. The finding that the pathogen-induced virulence phenotype affects the fate of S. enterica populations in diseased plants suggests that targeting of plant pathogen disease is important in controlling S. enterica populations on plants. PMID:26386057

  8. Salmonella serotypes in reptiles and humans, French Guiana.

    PubMed

    Gay, Noellie; Le Hello, Simon; Weill, François-Xavier; de Thoisy, Benoit; Berger, Franck

    2014-05-14

    In French Guiana, a French overseas territory located in the South American northern coast, nearly 50% of Salmonella serotypes isolated from human infections belong to serotypes rarely encountered in metropolitan France. A reptilian source of contamination has been investigated. Between April and June 2011, in the area around Cayenne, 151 reptiles were collected: 38 lizards, 37 snakes, 32 turtles, 23 green iguanas and 21 caimans. Cloacal swab samples were collected and cultured. Isolated Salmonella strains were identified biochemically and serotyped. The overall carriage frequency of carriage was 23.2% (95% confidence interval: 16.7-30.4) with 23 serotyped strains. The frequency of Salmonella carriage was significantly higher for wild reptiles. Near two-thirds of the Salmonella serotypes isolated from reptiles were also isolated from patients in French Guiana. Our results highlight the risk associated with the handling and consumption of reptiles and their role in the spread of Salmonella in the environment. PMID:24560590

  9. Presence and correlation of some enteric indicator bacteria, diarrheagenic Escherichia coli pathotypes, and Salmonella serotypes in alfalfa sprouts from local retail markets in Pachuca, Mexico.

    PubMed

    Rangel-Vargas, Esmeralda; Gómez-Aldapa, Carlos A; Torres-Vitela, M Del Refugio; Villarruel-López, Angélica; Gordillo-Martínez, Alberto J; Castro-Rosas, Javier

    2015-03-01

    Data on the presence of diarrheagenic Escherichia coli pathotypes (DEPs) in alfalfa sprouts and correlations between the presence of coliform bacteria (CB), fecal coliforms (FC), E. coli, DEPs, and Salmonella in alfalfa sprouts are not available. The presence of and correlations between CB, FC, E. coli, DEPs, and Salmonella in alfalfa sprouts were determined. One hundred sprout samples were collected from retail markets in Pachuca, Hidalgo State, Mexico. The presence of indicator bacteria and Salmonella was determined using conventional culture procedures. DEPs were identified using two multiplex PCR procedures. One hundred percent of samples were positive for CB, 90% for FC, 84% for E. coli, 10% for DEPs, and 4% for Salmonella. The populations of CB ranged from 6.2 up to 8.6 log CFU/g. The FC and E. coli concentrations were between , 3 and 1,100 most probable number (MPN)/g. The DEPs identified included enterotoxigenic E. coli (ETEC; 2%), enteropathogenic E. coli (EPEC; 3%), and Shiga toxin-producing E. coli (STEC; 5%). No E. coli O157:H7 strains were detected in any STEC-positive samples. In samples positive for DEPs, the concentrations ranged from 210 to 240 MPN/g for ETEC, 28 to 1,100 MPN/g for EPEC, and 3.6 to 460 MPN/g for STEC. The Salmonella isolates identified included Salmonella enterica serotype Typhimurium in three samples and Salmonella enterica serotype Enteritidis in one. STEC and Salmonella Typhimurium were identified together in one sample. Positive correlations were observed between FC and E. coli, between FC and DEPs, and between E. coli and DEPs. Negative correlations occurred between CB and DEPs and between CB and Salmonella. Neither FC nor E. coli correlated with Salmonella in the sprout samples. To our knowledge, this is the first report of ETEC, EPEC, and STEC isolated from alfalfa sprouts and the first report of correlations between different indicator groups versus DEPs and Salmonella. PMID:25719889

  10. Serotype-specific and serotype-independent strategies for pre-harvest control of foodborne Salmonella in poultry.

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Of more than 2500 identified Salmonella serotypes, only a small proportion are common in poultry flocks. However, there is an epidemiologically important connection between poultry products and human infections, as many of the serotypes that are most prevalent in humans (such as S. Typhimurium and S...

  11. Prevalence and Characterization of Salmonella enterica and Salmonella Bacteriophages Recovered from Beef Cattle Feedlots in South Texas.

    PubMed

    Xie, Yicheng; Savell, Jeffrey W; Arnold, Ashley N; Gehring, Kerri B; Gill, Jason J; Taylor, T Matthew

    2016-08-01

    Asymptomatic Salmonella carriage in beef cattle is a food safety concern, and the beef feedlot environment may function as a reservoir of this pathogen. The goal of this study was to identify and isolate Salmonella and Salmonella bacteriophages from beef cattle feedlot environments in order to better understand the microbial ecology of Salmonella and identify phages that might be useful as anti-Salmonella beef safety interventions. Three feedlots in south Texas were visited, and 27 distinct samples from each source were collected from dropped feces, feed from feed bunks, drinking water from troughs, and soil in cattle pens (n = 108 samples). Preenrichment, selective enrichment, and selective/differential isolation of Salmonella were performed on each sample. A representative subset of presumptive Salmonella isolates was prepared for biochemical identification and serotyping. Samples were pooled by feedlot and sample type to create 36 samples and enriched to recover phages. Recovered phages were tested for host range against two panels of Salmonella hosts. Salmonella bacteria were identified in 20 (18.5%) of 108 samples by biochemical and/or serological testing. The serovars recovered included Salmonella enterica serovars Anatum, Muenchen, Altona, Kralingen, Kentucky, and Montevideo; Salmonella Anatum was the most frequently recovered serotype. Phage-positive samples were distributed evenly over the three feedlots, suggesting that phage prevalence is not strongly correlated with the presence of culturable Salmonella. Phages were found more frequently in soil and feces than in feed and water samples. The recovery of bacteriophages in the Salmonella-free feedlot suggests that phages might play a role in suppressing the Salmonella population in a feedlot environment. PMID:27497120

  12. Molecular characterization of Salmonella enterica serovar Saintpaul isolated from imported seafood, pepper, environmental and clinical samples.

    PubMed

    Akiyama, Tatsuya; Khan, Ashraf A; Cheng, Chorng-Ming; Stefanova, Rossina

    2011-09-01

    A total of 39 Salmonella enterica serovar Saintpaul strains from imported seafood, pepper and from environmental and clinical samples were analyzed for the presence of virulence genes, antibiotic resistance, plasmid and plasmid replicon types. Pulsed-field gel electrophoresis (PFGE) fingerprinting using the XbaI restriction enzyme and plasmid profiling were performed to assess genetic diversity. None of the isolates showed resistance to ampicillin, chloramphenicol, gentamicin, kanamycin, streptomycin, sulfisoxazole, and tetracycline. Seventeen virulence genes were screened for by PCR. All strains were positive for 14 genes (spiA, sifA, invA, spaN, sopE, sipB, iroN, msgA, pagC, orgA, prgH, lpfC, sitC, and tolC) and negative for three genes (spvB, pefA, and cdtB). Twelve strains, including six from clinical samples and six from seafood, carried one or more plasmids. Large plasmids, sized greater than 50 kb were detected in one clinical and three food isolates. One plasmid was able to be typed as IncI1 by PCR-based replicon typing. There were 25 distinct PFGE-XbaI patterns, clustered to two groups. Cluster A, with 68.5% similarity mainly consists of clinical isolates, while Cluster C, with 67.6% similarity, mainly consisted of shrimp isolates from India. Our findings indicated the genetic diversity of S. Saintpaul in clinical samples, imported seafood, and the environment and that this serotype possesses several virulent genes and plasmids which can cause salmonellosis. PMID:21645810

  13. Requirement of siderophore biosynthesis for plant colonization by Salmonella enterica

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Contaminated fresh produce has become the number one vector of non-typhoidal salmonellosis to humans. However, Salmonella enterica genes essential for the life cycle of this organism outside the mammalian host are for the most part unknown. Screening deletion mutants led to the discovery that an aro...

  14. Limited Genetic Diversity in Salmonella enterica Serovar Enteritidis PT13

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Salmonella enterica serovar Enteritidis has emerged as a significant food-borne pathogen throughout the world and it is commonly characterized by phage typing (PT). In Canada, PT4, 8 and 13 are the predominant PTs. Epidemiological subtyping of Salmonella is typically done by PFGE but plasmid profil...

  15. Multidrug-Resistant Salmonella enterica Serovar Infantis, Israel

    PubMed Central

    Valinsky, Lea; Weinberger, Miriam; Guy, Sara; Jaffe, Joseph; Schorr, Yosef Ilan; Raisfeld, Abraham; Agmon, Vered; Nissan, Israel

    2010-01-01

    To determine whether rapid emergence of Salmonella enterica serovar Infantis in Israel resulted from an increase in different biotypes or spread of 1 clone, we characterized 87 serovar Infantis isolates on the genotypic and phenotypic levels. The emerging strain comprised 1 genetic clone with a distinct pulsed-field gel electrophoresis profile and a common antimicrobial drug resistance pattern. PMID:21029536

  16. Method for the detection of Salmonella enterica serovar Enteritidis

    DOEpatents

    Agron, Peter G.; Andersen, Gary L.; Walker, Richard L.

    2008-10-28

    Described herein is the identification of a novel Salmonella enterica serovar Enteritidis locus that serves as a marker for DNA-based identification of this bacterium. In addition, three primer pairs derived from this locus that may be used in a nucleotide detection method to detect the presence of the bacterium are also disclosed herein.

  17. Limited genetic diversity in Salmonella enterica Serovar Enteritidis PT13

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Salmonella enterica serovar Enteritidis has emerged as a significant foodborne pathogen throughout the world and is commonly characterized by phage typing. In Canada phage types (PT) 4, 8 and 13 predominate and in 2005 a large foodborne PT13 outbreak occurred in the province of Ontario. The ability ...

  18. Effect of residual sanitizers on Salmonella enterica biofilm formation

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Introduction: Salmonella enterica are a diverse group of bacteria that represent a serious risk to public health. Bacterial attachment on food and contact surfaces can lead to biofilm formation, and once in this state, bacteria are more resistant to sanitization and may serve as a continuous contam...

  19. Salmonella enterica Strains with Reduced Susceptibility to Quarternary Ammonium Compounds

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Background: Salmonella spp. are responsible for 76 million illnesses per year in the U.S. Quaternary ammonium compounds (QAC) are commonly used antimicrobial agents. Reduced susceptibility to these compounds by a broad spectrum of organisms is a concern. Methods: Salmonella enterica strains with r...

  20. Epidemiology of a Salmonella enterica subsp. enterica serovar Typhimurium strain associated with a songbird outbreak.

    PubMed

    Hernandez, Sonia M; Keel, Kevin; Sanchez, Susan; Trees, Eija; Gerner-Smidt, Peter; Adams, Jennifer K; Cheng, Ying; Ray, Al; Martin, Gordon; Presotto, Andrea; Ruder, Mark G; Brown, Justin; Blehert, David S; Cottrell, Walter; Maurer, John J

    2012-10-01

    Salmonella enterica subsp. enterica serovar Typhimurium is responsible for the majority of salmonellosis cases worldwide. This Salmonella serovar is also responsible for die-offs in songbird populations. In 2009, there was an S. Typhimurium epizootic reported in pine siskins in the eastern United States. At the time, there was also a human outbreak with this serovar that was associated with contaminated peanuts. As peanuts are also used in wild-bird food, it was hypothesized that the pine siskin epizootic was related to this human outbreak. A comparison of songbird and human S. Typhimurium pulsed-field gel electrophoresis (PFGE) patterns revealed that the epizootic was attributed not to the peanut-associated strain but, rather, to a songbird strain first characterized from an American goldfinch in 1998. This same S. Typhimurium strain (PFGE type A3) was also identified in the PulseNet USA database, accounting for 137 of 77,941 total S. Typhimurium PFGE entries. A second molecular typing method, multiple-locus variable-number tandem-repeat analysis (MLVA), confirmed that the same strain was responsible for the pine siskin epizootic in the eastern United States but was distinct from a genetically related strain isolated from pine siskins in Minnesota. The pine siskin A3 strain was first encountered in May 2008 in an American goldfinch and later in a northern cardinal at the start of the pine siskin epizootic. MLVA also confirmed the clonal nature of S. Typhimurium in songbirds and established that the pine siskin epizootic strain was unique to the finch family. For 2009, the distribution of PFGE type A3 in passerines and humans mirrored the highest population density of pine siskins for the East Coast. PMID:22885752

  1. Epidemiology of a Salmonella enterica subsp. Enterica serovar Typhimurium strain associated with a songbird outbreak.

    USGS Publications Warehouse

    Blehert, David S.; Hernandez, Sonia M.; Keel, Kevin; Sanchez, Susan; Trees, Eija; Peter Gerner-Smidt

    2012-01-01

    Salmonella enterica subsp. enterica serovar Typhimurium is responsible for the majority of salmonellosis cases worldwide. This Salmonella serovar is also responsible for die-offs in songbird populations. In 2009, there was an S. Typhimurium epizootic reported in pine siskins in the eastern United States. At the time, there was also a human outbreak with this serovar that was associated with contaminated peanuts. As peanuts are also used in wild-bird food, it was hypothesized that the pine siskin epizootic was related to this human outbreak. A comparison of songbird and human S. Typhimurium pulsed-field gel electrophoresis (PFGE) patterns revealed that the epizootic was attributed not to the peanut-associated strain but, rather, to a songbird strain first characterized from an American goldfinch in 1998. This same S. Typhimurium strain (PFGE type A3) was also identified in the PulseNet USA database, accounting for 137 of 77,941 total S. Typhimurium PFGE entries. A second molecular typing method, multiple-locus variable-number tandem-repeat analysis (MLVA), confirmed that the same strain was responsible for the pine siskin epizootic in the eastern United States but was distinct from a genetically related strain isolated from pine siskins in Minnesota. The pine siskin A3 strain was first encountered in May 2008 in an American goldfinch and later in a northern cardinal at the start of the pine siskin epizootic. MLVA also confirmed the clonal nature of S. Typhimurium in songbirds and established that the pine siskin epizootic strain was unique to the finch family. For 2009, the distribution of PFGE type A3 in passerines and humans mirrored the highest population density of pine siskins for the East Coast.

  2. Early immune response following Salmonella enterica subspecies enterica serovar Typhimurium infection in porcine jejunal gut loops.

    PubMed

    Meurens, François; Berri, Mustapha; Auray, Gael; Melo, Sandrine; Levast, Benoît; Virlogeux-Payant, Isabelle; Chevaleyre, Claire; Gerdts, Volker; Salmon, Henri

    2009-01-01

    Salmonella enterica subspecies enterica serovar Typhimurium, commonly called S. Typhimurium, can cause intestinal infections in humans and various animal species such as swine. To analyze the host response to Salmonella infection in the pig we used an in vivo gut loop model, which allows the analysis of multiple immune responses within the same animal. Four jejunal gut-loops were each inoculated with 3 x 10(8) cfu of S. Typhimurium in 3 one-month-old piglets and mRNA expressions of various cytokines, chemokines, transcription factors, antimicrobial peptides, toll like and chemokine receptors were assessed by quantitative real-time PCR in the Peyer's patch and the gut wall after 24 h. Several genes such as the newly cloned CCRL1/CCX-CKR were assessed for the first time in the pig at the mRNA level. Pro-inflammatory and T-helper type-1 (Th1) cytokine mRNA were expressed at higher levels in infected compared to non-infected control loops. Similarly, some B cell activation genes, NOD2 and toll like receptor 2 and 4 transcripts were more expressed in both tissues while TLR5 mRNA was down-regulated. Interestingly, CCL25 mRNA expression as well as the mRNA expressions of its receptors CCR9 and CCRL1 were decreased both in the Peyer's patch and gut wall suggesting a potential Salmonella strategy to reduce lymphocyte homing to the intestine. In conclusion, these results provide insight into the porcine innate mucosal immune response to infection with entero-invasive microorganisms such as S. Typhimurium. In the future, this knowledge should help in the development of improved prophylactic and therapeutic approaches against porcine intestinal S. Typhimurium infections. PMID:18922229

  3. Evaluation and comparison of molecular techniques for epidemiological typing of Salmonella enterica subsp. enterica serovar dublin.

    PubMed Central

    Liebisch, B; Schwarz, S

    1996-01-01

    A total of 28 unrelated isolates of the Salmonella enterica subsp. enterica serovar dublin (S. dublin) collected during a 6-year period, as well as four samples of the S. dublin live vaccine strain Bovisaloral and its prototype strain S. dublin 442/039, were investigated by different molecular typing methods for the following reasons: (i) to find the most discriminatory method for the epidemiological typing of isolates belonging to this Salmonella serovar and (ii) to evaluate these methods for their capacity to discriminate among the live vaccine strain Bovisaloral, its prototype strain S. dublin 442/039, and field isolates of the serovar dublin. Five different plasmid profiles were observed; a virulence plasmid of 76 kbp as identified by hybridization with an spvB-spvC gene probe was present in all isolates. The detection of 16S rRNA genes and that of IS200 elements proved to be unsuitable for the epidemiological typing of S. dublin; only one hybridization pattern could be observed with each of these methods. The results obtained from macrorestriction analysis strongly depended on the choice of restriction enzyme. While the enzyme NotI yielded the lowest discriminatory index among all enzymes tested, it was the only enzyme that allowed discrimination between the Bovisaloral vaccine strain and its prototype strain. In contrast to the enzymes XbaI and SpeI, which only differentiated among the S. dublin field isolates, XhoI as well as AvrII also produced restriction fragment patterns of the Bovisaloral strain and of its prototype strain that were not shared by any of the S. dublin field isolates. Macrorestriction analysis proved to be the most discriminatory method not only for the epidemiological typing of S. dublin field isolates but also for the identification of the S. dublin live vaccine strain Bovisaloral. PMID:8904430

  4. Global Distribution of Campylobacter jejuni Penner Serotypes: A Systematic Review

    PubMed Central

    Pike, Brian L.; Guerry, Patricia; Poly, Frédéric

    2013-01-01

    Penner serotyping has been the principal method for differentiating Campylobacter isolates since its inception. Campylobacter capsule polysaccharide (CPS), the principal serodeterminant on which Penner serotyping is based, is presently of interest as a vaccine component. To determine the required valency of an effective CPS-based vaccine, a comprehensive understanding of CPS distribution is needed. Because of the association between Penner serotype and CPS, we conducted a systematic review to estimate the frequency and distribution of Penner serotypes associated with cases of Campylobacteriosis. In total, more than 21,000 sporadic cases of C. jejuni cases were identified for inclusion. While regional variation exists, distribution estimates indicate that eight serotypes accounted for more than half of all sporadic diarrheal cases globally and three serotypes (HS4 complex, HS2, and HS1/44) were dominant inter-regionally as well as globally. Furthermore, a total of 17 different serotypes reached a representation of 2% or greater in at least one of the five regions sampled. While this review is an important first step in defining CPS distribution, these results make it clear that significant gaps remain in our knowledge. Eliminating these gaps will be critical to future vaccine development efforts. PMID:23826280

  5. Toxoplasma Serotype Is Associated With Development of Ocular Toxoplasmosis

    PubMed Central

    Shobab, Leila; Pleyer, Uwe; Johnsen, Joerdis; Metzner, Sylvia; James, Erick R.; Torun, N.; Fay, Michael P.; Liesenfeld, Oliver; Grigg, Michael E.

    2013-01-01

    Background. Worldwide, ocular toxoplasmosis (OT) is the principal cause of posterior uveitis, a severe, life-altering disease. A Toxoplasma gondii enzyme-linked immunoassay that detects strain-specific antibodies present in serum was used to correlate serotype with disease. Methods. Toxoplasma serotypes in consecutive serum samples from German uveitis patients with OT were compared with non-OT seropositive patients with noninfectious autoimmune posterior uveitis. OT patients were tested for association of parasite serotype with age, gender, location, clinical onset, size, visual acuity, or number of lesions (mean follow-up, 3.8 years) to determine association with recurrences. Results. A novel, nonreactive (NR) serotype was detected more frequently in serum samples of OT patients (50/114, 44%) than in non-OT patients (4/56, 7%) (odds ratio, 10.0; 95% confidence interval 3.4–40.8; P < .0001). Non-OT patients were predominantly infected with Type II strains (39/56; 70%), consistent with expected frequencies in Central Europe. Among OT patients, those with NR serotypes experienced more frequent recurrences (P = .037). Polymerase chain reaction detected parasite DNA in 8/60 OT aqueous humor specimens but failed to identify Type II strain alleles. Conclusions. Toxoplasma NR and Type II serotypes predominate in German OT patients. The NR serotype is associated with OT recurrences, underscoring the value of screening for management of disease. PMID:23878321

  6. Draft Genome Sequences of Streptococcus agalactiae Serotype Ia and III Isolates from Tilapia Farms in Thailand

    PubMed Central

    Areechon, Nontawith; Kannika, Korntip; Hirono, Ikuo

    2016-01-01

    Streptococcus agalactiae serotypes Ia and III were isolated from infected tilapia in cage and pond culture farms in Thailand during 2012 to 2014, in which pathogenicity analysis demonstrated that serotype III showed higher virulence than serotype Ia. Here, we report the draft genome sequencing of piscine S. agalactiae serotypes Ia and III. PMID:27013037

  7. Draft Genome Sequence of a Salmonella enterica subsp. enterica Serovar Gallinarum bv. Gallinarum Isolate Associated with Fowl Typhoid Outbreaks in Brazil

    PubMed Central

    De Carli, Silvia; Gräf, Tiago; Mayer, Fabiana Q.; Cibulski, Samuel; Lehmann, Fernanda K. M.; Fonseca, André S. K.; Ikuta, Nilo

    2016-01-01

    Salmonella enterica subsp. enterica serovar Gallinarum bv. Gallinarum strains are bird pathogens causing fowl typhoid (FT). Isolate BR_RS12 was obtained from a poultry flock with FT in 2014. The sequencing of this genome will enable to track the origin of the recent outbreaks in Brazil. PMID:26950322

  8. Draft Genome Sequence and Annotation of Phyllosphere-Persisting Salmonella enterica subsp. enterica Serovar Livingstone Strain CKY-S4, Isolated from an Urban Lake in Regina, Canada.

    PubMed

    Tambalo, Dinah D; Perry, Benjamin J; Fitzgerald, Stephen F; Cameron, Andrew D S; Yost, Christopher K

    2015-01-01

    Here, we report the first draft genome sequence of Salmonella enterica subsp. enterica serovar Livingstone. This S. Livingstone strain CKY-S4 displayed biofilm formation and cellulose production and could persist on lettuce. This genome may help the study of mechanisms by which enteric pathogens colonize food crops. PMID:26272568

  9. Draft Genome Sequence and Annotation of Phyllosphere-Persisting Salmonella enterica subsp. enterica Serovar Livingstone Strain CKY-S4, Isolated from an Urban Lake in Regina, Canada

    PubMed Central

    Tambalo, Dinah D.; Perry, Benjamin J.; Fitzgerald, Stephen F.; Cameron, Andrew D. S.

    2015-01-01

    Here, we report the first draft genome sequence of Salmonella enterica subsp. enterica serovar Livingstone. This S. Livingstone strain CKY-S4 displayed biofilm formation and cellulose production and could persist on lettuce. This genome may help the study of mechanisms by which enteric pathogens colonize food crops. PMID:26272568

  10. Draft Genome Sequence of Salmonella enterica subsp. enterica Serovar Napoli Strain SN310, Cause of a Multischool Outbreak in Milan, Italy, in 2014.

    PubMed

    Huedo, Pol; Gori, Maria; Scaltriti, Erika; Morganti, Marina; Casadei, Gabriele; Amato, Ettore; Pontello, Mirella

    2015-01-01

    We report the draft genome sequence of Salmonella enterica subsp. enterica serovar Napoli strain SN310, isolated from a stool sample of an affected pupil during a multischool outbreak in 2014 in Milan, Italy. This represents the first reported draft genome sequence of the emerging serovar Napoli. PMID:26358605

  11. Enteropathogenic Escherichia coli Serotypes and Endemic Diarrhea in Infants

    PubMed Central

    Toledo, M. Regina F.; Alvariza, M. do Carmo B.; Murahovschi, Jayme; Ramos, Sonia R. T. S.; Trabulsi, Luiz R.

    1983-01-01

    Enteropathogenic Escherichia coli serotypes were searched for in feces of 550 children with endemic diarrhea and in 129 controls, in São Paulo, in 1978 and 1979; serotypes O111ab:H−, O111ab:H2, and O119:H6 were significantly associated with diarrhea in children 0 to 5 months old and were the most frequent agents of diarrhea in this age group as compared with enterotoxigenic and enteroinvasive E. coli, Salmonella sp., Shigella sp., and Yersinia enterocolitica. It is concluded that various enteropathogenic E. coli serotypes may be agents of endemic infantile diarrhea. PMID:6339384

  12. Complete and closed genome sequences of 10 Salmonella enterica subsp. enterica serovar Anatum isolated from human and bovine sources

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Salmonella enterica is an important pathogen transmitted by numerous vectors. Genomic comparisons of Salmonella from disparate hosts have the potential to further our understanding of mechanisms underlying host specificities and virulence. Here, we present closed genome and plasmid sequences of 10...

  13. SALMATcor: microagglutination for Salmonella flagella serotyping.

    PubMed

    Duarte Martínez, Francisco; Sánchez-Salazar, Luz Marina; Acuña-Calvo, María Teresa; Bolaños-Acuña, Hilda María; Dittel-Dittel, Isis; Campos-Chacón, Elena

    2010-08-01

    Salmonella is a complex bacterial group with more than 2400 serovars widely distributed in nature; they are considered zoonotic because they can infect a variety of animals and be transmitted to humans. Usually, they cause alimentary acquired diseases such as gastroenteritis, typhoid fever, and others that can lead to severe complications and death. Serotyping is useful to differentiate among Salmonella, because it shows an important correlation with their clinical and epidemiological patterns; consequently, it is of high value for public health, animal health, agriculture, and industry. To characterize all known Kauffmann-White Salmonella serovars, over 250 antisera are required. Due to this and to high prices antisera, many laboratories worldwide have limitations in establishing Salmonella surveillance. Therefore, we developed and validated a Salmonella flagella microagglutination test (SALMATcor) that significantly reduces laboratory requirements of antisera. SALMATcor is based on scaling down, by fivefold, the antigen:antiserum volumes actually required for the reference method: flagella standard tube agglutination technique (STAT). Antigen preparation, temperatures, and incubation periods remained as established for STAT. The SALMATcor was validated according to ISO/DIS 16140:1999 protocol, which included 1187 comparisons of flagella determinations conducted by SALMATcor and STAT, on 141 Salmonella isolates of 12 common serotypes and the use of antiserum recommended for STAT. SALMATcor concordance was excellent (Cohen's kappa index 0.9982), obtaining relative accuracy >99.9% and relative specificity >99.9%. Additionally, SALMATcor has been used by CNRB-INCIENSA since 2004 to respond to all 40 Salmonella proficiency testing strains, provided by World Health Organization-Global Salmonella Surveillance Network, obtaining 100% concordance on serovar identification. On the basis of the results achieved with SALMATcor and considering that it also significantly

  14. Prevalence of shiga toxin producing Escherichia coli, Salmonella enterica, and Listeria monocytogenes at public access watershed sites in a California Central Coast agricultural region.

    PubMed

    Cooley, Michael B; Quiñones, Beatriz; Oryang, David; Mandrell, Robert E; Gorski, Lisa

    2014-01-01

    Produce contaminated with enteric pathogens is a major source of foodborne illness in the United States. Lakes, streams, rivers, and ponds were sampled with Moore swabs bi-monthly for over 2 years at 30 locations in the vicinity of a leafy green growing region on the Central California Coast and screened for Shiga toxin producing Escherichia coli (STEC), Salmonella enterica, and Listeria monocytogenes to evaluate the prevalence and persistence of pathogen subtypes. The prevalence of STEC from 1386 samples was 11%; 110 samples (8%) contained E. coli O157:H7 with the highest prevalence occurring close to cattle operations. Non-O157 STEC isolates represented major clinical O-types and 57% contained both shiga toxin types 1 and 2 and intimin. Multiple Locus Variable Number Tandem Repeat Analysis of STEC isolates indicated prevalent strains during the period of study. Notably, Salmonella was present at high levels throughout the sampling region with 65% prevalence in 1405 samples resulting in 996 isolates with slightly lower prevalence in late autumn. There were 2, 8, and 14 sites that were Salmonella-positive over 90, 80, and 70% of the time, respectively. The serotypes identified most often were 6,8:d:-, Typhimurium, and Give. Interestingly, analysis by Pulsed Field Gel Electrophoresis indicated persistence and transport of pulsotypes in the region over several years. In this original study of L. monocytogenes in the region prevalence was 43% of 1405 samples resulting in 635 individual isolates. Over 85% of the isolates belonged to serotype 4b with serotypes 1/2a, 1/2b, 3a, 4d with 4e representing the rest, and there were 12 and 2 sites that were positive over 50 and 80% of the time, respectively. Although surface water is not directly used for irrigation in this region, transport to the produce can occur by other means. This environmental survey assesses initial contamination levels toward an understanding of transport leading to produce recalls or outbreaks. PMID

  15. Prevalence of shiga toxin producing Escherichia coli, Salmonella enterica, and Listeria monocytogenes at public access watershed sites in a California Central Coast agricultural region

    PubMed Central

    Cooley, Michael B.; Quiñones, Beatriz; Oryang, David; Mandrell, Robert E.; Gorski, Lisa

    2014-01-01

    Produce contaminated with enteric pathogens is a major source of foodborne illness in the United States. Lakes, streams, rivers, and ponds were sampled with Moore swabs bi-monthly for over 2 years at 30 locations in the vicinity of a leafy green growing region on the Central California Coast and screened for Shiga toxin producing Escherichia coli (STEC), Salmonella enterica, and Listeria monocytogenes to evaluate the prevalence and persistence of pathogen subtypes. The prevalence of STEC from 1386 samples was 11%; 110 samples (8%) contained E. coli O157:H7 with the highest prevalence occurring close to cattle operations. Non-O157 STEC isolates represented major clinical O-types and 57% contained both shiga toxin types 1 and 2 and intimin. Multiple Locus Variable Number Tandem Repeat Analysis of STEC isolates indicated prevalent strains during the period of study. Notably, Salmonella was present at high levels throughout the sampling region with 65% prevalence in 1405 samples resulting in 996 isolates with slightly lower prevalence in late autumn. There were 2, 8, and 14 sites that were Salmonella-positive over 90, 80, and 70% of the time, respectively. The serotypes identified most often were 6,8:d:-, Typhimurium, and Give. Interestingly, analysis by Pulsed Field Gel Electrophoresis indicated persistence and transport of pulsotypes in the region over several years. In this original study of L. monocytogenes in the region prevalence was 43% of 1405 samples resulting in 635 individual isolates. Over 85% of the isolates belonged to serotype 4b with serotypes 1/2a, 1/2b, 3a, 4d with 4e representing the rest, and there were 12 and 2 sites that were positive over 50 and 80% of the time, respectively. Although surface water is not directly used for irrigation in this region, transport to the produce can occur by other means. This environmental survey assesses initial contamination levels toward an understanding of transport leading to produce recalls or outbreaks. PMID

  16. A Multischool Outbreak Due to Salmonella enterica serovar Napoli Associated with Elevated Rates of Hospitalizations and Bacteremia, Milan, Italy, 2014.

    PubMed

    Huedo, Pol; Gori, Maria; Amato, Ettore; Bianchi, Roberta; Valerio, Edgardo; Magnoli, Luigi; Pontello, Mirella

    2016-08-01

    A multischool outbreak of salmonellosis caused by Salmonella enterica serovar Napoli was investigated in the province of Milan from October to November 2014, following an increase in school absenteeism coinciding with two positive cases. Epidemiological studies detected 47 cases in four primary schools: 46 children and 1 adult woman (51.4% males and 48.6% females, median age 8.9). From these, 14 cases (29.8%) were severe and resulted in hospitalization, including 6 children (12.8%) who developed an invasive salmonellosis. The epidemic curve revealed an abnormally long incubation period, peaking 1 week after the first confirmed case. Twenty-five available isolates were typed by pulsed-field gel electrophoresis showing an identical pattern. The isolate belongs to ST474, an ST composed exclusively of Salmonella Napoli human strains isolated in France and Italy. Antibiotic resistance analysis showed resistance to aminoglycosides, correlating with the presence of the aminoglycoside resistance gene aadA25 in its genome. Trace-back investigations strongly suggested contaminated ham as the most likely food vehicle, which was delivered by a common food center on 21 October. Nevertheless, this ingredient could not be retrospectively investigated since it was no longer available at the repository. This represents the largest Salmonella Napoli outbreak ever reported in Italy and provides a unique scenario for studying the outcome of salmonellosis caused by this emerging and potentially invasive nontyphoidal serotype. PMID:27148636

  17. Core-linked LPS expression of Shigella dysenteriae serotype 1 O-antigen in live Salmonella Typhi vaccine vector Ty21a: preclinical evidence of immunogenicity and protection.

    PubMed

    Xu, De Qi; Cisar, John O; Osorio, Manuel; Wai, Tint T; Kopecko, Dennis J

    2007-08-14

    Shigella dysenteriae serotype 1 (S. dysenteriae 1) causes severe shigellosis that is typically associated with high mortality. Antibodies against Shigella serotype-specific O-polysaccharide (O-Ps) have been shown to be host protective. In this study, the rfb locus and the rfp gene with their cognate promoter regions were PCR-amplified from S. dysenteriae 1, cloned, and sequenced. Deletion analysis showed that eight rfb ORFs plus rfp are necessary for biosynthesis of this O-Ps. A tandemly-linked rfb-rfp gene cassette was cloned into low copy plasmid pGB2 to create pSd1. Avirulent Salmonella enterica serovar Typhi (S. Typhi) Ty21a harboring pSd1 synthesized S. Typhi 9, 12 LPS as well as typical core-linked S. dysenteriae 1 LPS. Animal immunization studies showed that Ty21a (pSd1) induces protective immunity against high stringency challenge with virulent S. dysenteriae 1 strain 1617. These data further demonstrate the utility of S. Typhi Ty21a as a live, bacterial vaccine delivery system for heterologous O-antigens, supporting the promise of a bifunctional oral vaccine for prevention of shigellosis and typhoid fever. PMID:17629369

  18. Fungal serotype-specific differences in bacterial-yeast interactions

    PubMed Central

    Abdulkareem, Asan F; Lee, Hiu Ham; Ahmadi, Mohammed; Martinez, Luis R

    2015-01-01

    Cryptococcus neoformans (Cn) causes meningoencephalitis in immunocompromised individuals. This encapsulated fungus can be found interacting with environmental microbes in soil contaminated with pigeon excrement. Cn survival within polymicrobial and other challenging communities has been shown to affect the evolution of its virulence factors. We compared the survival of 10 serotype A and D strains after interaction with the soil bacterium, Acinetobacter baumannii (Ab). Although co-incubation with Ab stimulated virulence factors production by strains of both cryptococcal serotypes, on average, serotype A strains displayed significantly higher survival rate, number of metabolically active cells within biofilms, and capsular polysaccharide production and release than serotype D strains. Our findings suggest that interactions of Cn with other microorganisms influence the fungus' regulation and production of virulence factors, important elements needed for the successful colonization of the human host. PMID:26132337

  19. [Investigation of pathogenic phenotypes and virulence determinants of food-borne Salmonella enterica strains in Caenorhabditis elegans animal model].

    PubMed

    Aksoy, Deniz; Şen, Ece

    2015-10-01

    Salmonellosis, caused by non-typhoidal Salmonella enterica serovars with the consumption of contaminated food, is one of the leading food-borne disease that makes microbial food safety an important public health issue. This study was performed in order to determine the antibiotic resistance, serotyping, plasmid profiles and pathogenicity potentials of food-borne Salmonella isolates in Caenorhabditis elegans animal model system in Edirne province, located at Thrace region of Turkey. In this study, 32 Salmonella isolates, of which 26 belonged to Infantis, four to Enteritidis, one to Telaviv and one to Kentucky serovars, isolated from chicken carcasses were used. Antibiotic resistance profiles were determined by disc diffusion and broth microdilution methods. A new C.elegans nematode animal model system was used to determine the pathogenicity potential of the isolates. The antibiotic resistance profiles revealed that one (3.1%) isolate was resistant to gentamicin, two (6.2%) to ciprofloxacin, three (9.4%) to ampicillin, 18 (56.3%) to kanamycin, 19 (60.8%) to neomycin, 25 (78.1%) to tetracycline, 25 (78.1%) to trimethoprim, 26 (81.25%) to nalidixic acid, 27 (84.4%) to streptomycin and 32 (100%) to sulfonamide. All of the 32 strains were susceptible to chloramphenicol and ampicillin/sulbactam. High levels of resistance to streptomycin, nalidixic acid, tetracycline, trimethoprim, sulfonamide, kanamycin and neomycin was determined. According to the plasmid analysis, six isolates (18.75%) harboured 1-3 plasmids with sizes between 1.2 and 42.4 kb. In C.elegans nematode animal model system, the time (in days) required to kill 50% (TD50) of nematodes was calculated for each experimental group. TD50 values of the nematode group fed with S.Typhimurium ATCC 14028 that was used as the positive control and another group fed with E.coli OP50 as the negative control were 4.2 ± 0.5 days and 8.0 ± 0.02 days, respectively. TD50 of the groups fed with Salmonella isolates ranged

  20. Purification and characterization of a protease from Actinobacillus pleuropneumoniae serotype 1, an antigen common to all the serotypes.

    PubMed Central

    Negrete-Abascal, E; Tenorio, V R; Guerrero, A L; García, R M; Reyes, M E; de la Garza, M

    1998-01-01

    A high molecular-mass proteolytic enzyme of Actinobacillus pleuropneumoniae serotype 1, was purified from culture supernatants (CSN) by using DEAE-cellulose and sepharose-4B-gelatin chromatography. In 10% SDS-polyacrylamide gels copolymerized with porcine gelatin, the protease showed a single band of activity of > 200 kDa. However, minor molecular-mass proteolytic bands were observed when the protease was electrophoresed in the presence of either 5% beta-mercaptoethanol, 50 mM dithiothreitol, or 0.25 M urea. Furthermore, when the > 200-kDa purified protein was passed through a sucrose gradient, several bands with proteolytic activity were found: 62, 90, 190, and 540 kDa. The proteolytic activity was increased in the presence of calcium or zinc and was not affected after being heated at 90 degrees C for 5 min. Proteolytic activities were also observed in CSN from all A. pleuropneumoniae serotypes and biotypes. The purified protease hydrolyzed porcine IgA and IgG in vitro. In addition, by immunoblot the protease was recognized by serum of naturally infected pigs with serotypes 1 and 5, and by serum of pigs experimentally infected with serotypes 1, 2, 8, or 9. Serum of a pig vaccinated with CSN of a serotype 3 strain also recognized the protease, but not sera of pigs vaccinated with a bacterin (serotype 1). Proteins from CSN of all the serotypes, which were precipitated with 70% (NH4)2SO4, were recognized by a polyclonal antibody raised against the purified protease. Taken together these results indicate that an antigenic protease is produced in vivo by all the serotypes of A. pleuropneumoniae. The results indicate that proteases could have a role in the disease and in the immune response of pigs infected with A. pleuropneumoniae. Images Figure 2A. Figure 2B. Figure 3. Figure 4. Figure 5A. Figure 5B. Figure 6A. Figure 6B. PMID:9684047

  1. Quantification of the Sensitivity of Mycobacterium avium subsp paratuberculosis and Salmonella enterica subsp enterica to Low pH and High Organic Acids using Propidium Monoazide and Quantitative PCR

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Mycobacterium avium subsp paratuberculosis (Map) and Salmonella enterica subsp enterica (S. enterica) are two pathogens that are a concern to food and animal safety due to their ability to withstand harsh conditions encountered in the natural environment and within the host during pathogenesis. Acid...

  2. Serotyping of Campylobacter jejuni isolated from sporadic cases and outbreaks in British Columbia.

    PubMed Central

    McMyne, P M; Penner, J L; Mathias, R G; Black, W A; Hennessy, J N

    1982-01-01

    Campylobacter jejuni from sporadic cases and outbreaks of gastroenteritis were serotyped on the basis of heat-extracted soluble thermostable antigens identified with the use of the passive hemagglutination technique. A total of 168 isolates were separated into 45 different types. The largest proportion of the isolates fell into three serotypes, each with 11 to 12.5% of the total number. Three less frequently occurring serotypes each included approximately 5%, and the remaining 50% of the isolates were distributed among 39 other serotypes. In most cases, serotyping demonstrated that epidemiologically linked isolates were of the same serotype, but the outbreak strains could belong either to frequently or to infrequently isolated serotypes. The high correlation between clinical findings and serotyping results confirmed the applicability of the serotyping scheme in epidemiological investigations of C. jejuni infections. PMID:7119100

  3. E-nose identification of Salmonella enterica in poultry manure.

    PubMed

    Kizil, Ü; Genç, L; Genç, T T; Rahman, S; Khaitsa, M L

    2015-04-01

    A DiagNose II electronic nose (e-nose) system was tested to evaluate the performance of such systems in the detection of the Salmonella enterica pathogen in poultry manure. To build a database, poultry manure samples were collected from 7 broiler houses, samples were homogenised, and subdivided into 4 portions. One portion was left as is; the other three portions were artificially infected with S. enterica. An artificial neural network (ANN) model was developed and validated using the developed database. In order to test the performance of DiagNose II and the ANN model, 16 manure samples were collected from 6 different broiler houses and tested using these two systems. The results showed that DiagNose II was able to classify manure samples correctly as infected or non-infected based on the ANN model developed with a 94% level of accuracy. PMID:25650129

  4. Quantitative Oligonucleotide Microarray Fingerprinting of Salmonella enterica isolates

    SciTech Connect

    Willse, Alan R.; Straub, Tim M.; Wunschel, Sharon C.; Small, Jack A.; Call, Douglas R.; Daly, Don S.; Chandler, Darrell P.

    2004-03-22

    We report on a genome-independent microbial fingerprinting method using nucleic acid microarrays for microbial forensics and epidemiology applications. We demonstrate that the microarray method provides high-resolution differentiation between closely related microorganisms using Salmonella enterica strains. In replicate trials we used a simple 192-probe nonamer array to construct a fingerprint library of 25 closely related Salmonella isolates. Controlling false discovery rate for multiple testing at alpha =.05, at least 295 of 300 pairs of S. enterica isolate fingerprints were found to be statistically distinct using a modified Hotelling Tsquared test. Although we find most pairs of Salmonella fingerprints to be distinct, forensic applications will also require a protocol for library construction and reliable microbial classification against a fingerprint library. We outline additional steps required to produce a protocol for library construction and reliable classification of unknown organisms.

  5. Transcriptional profile of Salmonella enterica subsp. enterica serovar Weltevreden during alfalfa sprout colonization

    PubMed Central

    Brankatschk, Kerstin; Kamber, Tim; Pothier, Joël F; Duffy, Brion; Smits, Theo H M

    2014-01-01

    Sprouted seeds represent a great risk for infection by human enteric pathogens because of favourable growth conditions for pathogens during their germination. The aim of this study was to identify mechanisms of interactions of Salmonella enterica subsp. enterica Weltevreden with alfalfa sprouts. RNA-seq analysis of S. Weltevreden grown with sprouts in comparison with M9-glucose medium showed that among a total of 4158 annotated coding sequences, 177 genes (4.3%) and 345 genes (8.3%) were transcribed at higher levels with sprouts and in minimal medium respectively. Genes that were higher transcribed with sprouts are coding for proteins involved in mechanisms known to be important for attachment, motility and biofilm formation. Besides gene expression required for phenotypic adaption, genes involved in sulphate acquisition were higher transcribed, suggesting that the surface on alfalfa sprouts may be poor in sulphate. Genes encoding structural and effector proteins of Salmonella pathogenicity island 2, involved in survival within macrophages during infection of animal tissue, were higher transcribed with sprouts possibly as a response to environmental conditions. This study provides insight on additional mechanisms that may be important for pathogen interactions with sprouts. PMID:24308841

  6. Chromosome-Mediated Multidrug Resistance in Salmonella enterica Serovar Typhi

    PubMed Central

    Alam, Munirul; Kuo, Jung-Che; Liu, Yen-Yi; Wang, Pei-Jen

    2014-01-01

    A salmonella genomic island, designated SGI11, was found in 18 of 26 multidrug-resistant Salmonella enterica serovar Typhi isolates from Bangladesh. SGI11 was an IS1 composite transposon and carried 7 resistance genes that conferred resistance to 5 first-line antimicrobials. Eleven of the 18 SGI11-carrying S. Typhi isolates had developed resistance to high levels of ciprofloxacin. PMID:25367917

  7. Regulation of the ansB gene of Salmonella enterica.

    PubMed

    Jennings, M P; Scott, S P; Beacham, I R

    1993-07-01

    The expression of L-asparaginase II (encoded by ansB) in Salmonella enterica was found to be positively regulated by the cAMP receptor protein (CRP) and anaerobiosis. The anaerobic regulation of the S. enterica ansB gene is not mediated by the anaerobic transcriptional activator FNR. This is unlike the situation of the ansB gene of Escherichia coli, which is dependent on both CRP and FNR. To investigate this fundamental difference in the regulation of L-asparaginase II expression in S. enterica, the ansB gene was cloned and the nucleotide sequence of the promoter region determined. Sequence analysis and transcript mapping of the 5' promoter region revealed a single transcriptional start point (tsp) and two regulatory sites with substantial homology with those found in E. coli. One site, centred -90.5 bp from the tsp, is homologous to a hybrid CRP/FNR ('CF') site which is the site of CRP regulation in the E. coli promoter. The other site, centred 40.5 bp upstream of the tsp, is homologous to the FNR binding site of the E. coli promoter. Significantly, however, a single base-pair difference exists in this site, at a position of the related CRP and FNR DNA-binding site consensus sequences known to be involved in CRP versus FNR specificity. Site-directed mutagenesis indicates that this single difference, relative to the homologous E. coli site, results in a CRP binding site and the observed FNR-independent ansB expression in S. enterica.(ABSTRACT TRUNCATED AT 250 WORDS) PMID:8412661

  8. Antimicrobial Susceptibility and Plasmid Replicon Typing of Salmonella enterica serovar Kentucky isolates recovered from Broilers

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Salmonella Kentucky has become the predominate serotype recovered from broiler slaughter in the United States and the prevalence of antimicrobial resistance (AMR) has increased dramatically in this serotype. Relationships between AMR, genotype, and plasmid replicon types were characterized for 600 ...

  9. Mobilome differences between Salmonella enterica serovars Anatum and Typhimurium isolated from cattle and humans and potential impact on virulence

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Salmonella enterica subsp. enterica is an important group of pathogens capable of inhabiting a range of niches and hosts with varying degrees of impact, from commensal colonization to invasive infection. Recent outbreaks of multi-drug resistant S. enterica, attributed to consumption of contaminated ...

  10. A Highly Effective Component Vaccine against Nontyphoidal Salmonella enterica Infections

    PubMed Central

    Ferreira, Rosana B. R.; Valdez, Yanet; Coombes, Brian K.; Sad, Subash; Gouw, Joost W.; Brown, Eric M.; Li, Yuling; Grassl, Guntram A.; Antunes, L. Caetano M.; Gill, Navkiran; Truong, Mimi; Scholz, Roland; Reynolds, Lisa A.; Krishnan, Laskshmi; Zafer, Ahmed A.; Sal-Man, Neta; Lowden, Michael J.; Auweter, Sigrid D.; Foster, Leonard J.

    2015-01-01

    ABSTRACT Nontyphoidal Salmonella enterica (NTS) infections are a major burden to global public health, as they lead to diseases ranging from gastroenteritis to systemic infections and there is currently no vaccine available. Here, we describe a highly effective component vaccine against S. enterica serovar Typhimurium in both gastroenteritis and systemic murine infection models. We devised an approach to generate supernatants of S. enterica serovar Typhimurium, an organism that is highly abundant in virulence factors. Immunization of mice with this supernatant resulted in dramatic protection against a challenge with serovar Typhimurium, showing increased survival in the systemic model and decreased intestinal pathology in the gastrointestinal model. Protection correlated with specific IgA and IgG levels in the serum and specific secretory IgA levels in the feces of immunized mice. Initial characterization of the protective antigens in the bacterial culture supernatants revealed a subset of antigens that exhibited remarkable stability, a highly desirable characteristic of an effective vaccine to be used under suboptimal environmental conditions in developing countries. We were able to purify a subset of the peptides present in the supernatants and show their potential for immunization of mice against serovar Typhimurium resulting in a decreased level of colonization. This component vaccine shows promise with regard to protecting against NTS, and further work should significantly help to establish vaccines against these prevalent infections. PMID:26396246

  11. Selective and Genetic Constraints on Pneumococcal Serotype Switching

    PubMed Central

    Croucher, Nicholas J.; Kagedan, Lisa; Thompson, Claudette M.; Parkhill, Julian; Bentley, Stephen D.; Finkelstein, Jonathan A.; Lipsitch, Marc; Hanage, William P.

    2015-01-01

    Streptococcus pneumoniae isolates typically express one of over 90 immunologically distinguishable polysaccharide capsules (serotypes), which can be classified into “serogroups” based on cross-reactivity with certain antibodies. Pneumococci can alter their serotype through recombinations affecting the capsule polysaccharide synthesis (cps) locus. Twenty such “serotype switching” events were fully characterised using a collection of 616 whole genome sequences from systematic surveys of pneumococcal carriage. Eleven of these were within-serogroup switches, representing a highly significant (p < 0.0001) enrichment based on the observed serotype distribution. Whereas the recombinations resulting in between-serogroup switches all spanned the entire cps locus, some of those that caused within-serogroup switches did not. However, higher rates of within-serogroup switching could not be fully explained by either more frequent, shorter recombinations, nor by genetic linkage to genes involved in β–lactam resistance. This suggested the observed pattern was a consequence of selection for preserving serogroup. Phenotyping of strains constructed to express different serotypes in common genetic backgrounds was used to test whether genotypes were physiologically adapted to particular serogroups. These data were consistent with epistatic interactions between the cps locus and the rest of the genome that were specific to serotype, but not serogroup, meaning they were unlikely to account for the observed distribution of capsule types. Exclusion of these genetic and physiological hypotheses suggested future work should focus on alternative mechanisms, such as host immunity spanning multiple serotypes within the same serogroup, which might explain the observed pattern. PMID:25826208

  12. Structures of the SEp22 dodecamer, a Dps-like protein from Salmonella enterica subsp. enterica serovar Enteritidis

    PubMed Central

    Miyamoto, Takanori; Asahina, Yasuko; Miyazaki, Shohei; Shimizu, Hidetoshi; Ohto, Umeharu; Noguchi, Shuji; Satow, Yoshinori

    2011-01-01

    The crystal structure of SEp22, a DNA-binding protein from starved cells from Salmonella enterica subsp. enterica serovar Enteritidis, has been determined in two forms: the native state at 1.25 Å resolution and an iron-soaked form at 1.30 Å resolution. The SEp22 protomers form a dodecameric shell with 23 symmetry and a single iron ion per protomer was found at the ferroxidase centre in the iron-soaked form. Along the threefold axes of the 23 symmetry, hydrophilic Asp channels that consist of Asp146 were found. Iron ions may flow into the cavity of the dodecameric shell through the Asp channels. PMID:21206015

  13. Antibiotic resistant Escherichia coli and Salmonella enterica in the beef production and processing chain

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Background: Concerns have been raised that extended-spectrum cephalosporin-resistant Escherichia coli (CefR EC), trimethoprim-sulfamethoxazole-resistant E. coli (TxsR EC), extended-spectrum cephalosporin-resistant Salmonella enterica (CefR SE), and nalidixic acid-resistant S. enterica (NalR SE) in c...

  14. Genome-scale screening and validation of targets for identification of Salmonella enterica and serovar prediction

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Salmonella enterica is the most common foodborne pathogen worldwide, with a great diversity of 2500 recognized serovars. Detection of S. enterica and its classification into serovars are essential for food safety surveillance and clinical diagnosis. Recently, the polymerase chain reaction (PCR) meth...

  15. Improvements to a PCR-based serogrouping scheme for Salmonella enterica from dairy farm samples

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The PCR method described by Herrera-León, et al. (Research in Microbiology 158:122-127, 2007) has proved to be a simple and useful technique for characterizing isolates of Salmonella enterica enterica belonging to serogroups B, C1, C2, D1, and E1, groups which encompass a majority of the isolates fr...

  16. Role of Soil, Crop Debris, and a Plant Pathogen in Salmonella enterica Contamination of Tomato Plants

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Background: In the U.S., tomatoes have become the most implicated vehicle for produce-associated Salmonellosis with 12 outbreaks since 1998. Although unconfirmed, trace backs suggest pre-harvest contamination with Salmonella enterica. Routes of tomato crop contamination by S. enterica in the absence...

  17. [Fluorescent and Magnetic Relaxation Switch Immunosensor for the Detecting Foodborne Pathogen Salmonella enterica in Water Samples].

    PubMed

    Wang, Song-bai; Zhang, Yan; An, Wen-ting; Wei, Yan-li; Wang, Yu; Shuang, Shao-min

    2015-11-01

    Fluoroimmunoassay based on quantum dots (QDs) and magnetic relaxation switch (MRS) immunoassay based on superparamagnetic nanoparticles (SMN) were constructed to detect Salmonella enterica (S. enterica) in water samples. In fluoroimmunoassay, magnetic beads was conjugated with S. enterica capture antibody (MB-Ab2) to enrich S. enterica from sample solution, then the QDs was conjugated with the S. enterica detection antibody (QDs-Ab1) to detect S. enterica based on sandwich immunoassay format. And the fluorescence intensity is positive related to the bacteria concentration of the sample. Results showed that the limit of detection (LOD) of this method was 102 cfu · mL⁻¹ and analysis time was 2 h. In MRS assay, magnetic nanoparticle-antibody conjugate (MN-Ab1) can switch their dispersed and aggregated state in the presence of the target. This state of change can modulate the spin-spin relaxation time (T₂) of the neighboring water molecule. The change in T₂(ΔT₂) positively correlates with the amount of the target in the sample. Thus, AT can be used as a detection signal in MRS immunosensors. Results showed that LOD of MRS sensor for S. enterica was 10³ cfu · mL⁻¹ and analysis time was 0.5 h. Two methods were compared in terms of advantages and disadvantages in detecting S. enterica. PMID:26978918

  18. Studies on Biofilm Formation and Interactions of Salmonella enterica with Romaine-Lettuce Leaves

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The association between biofilm formation and the interactions of Salmonella enterica serovars with cut-Romaine-lettuce leaves was investigated. Biofilm formation by 8 S. enterica serovars was tested on polystyrene microtiter plates in the presence of different growth media. Maximal biofilm mass was...

  19. Previously uncharacterized Salmonella enterica genes required for swarming play a role in seedling colonization

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Incidences of bacterial foodborne illness caused by ingestion of fresh produce are rising. Instead of being a case of incidental contamination, the animal pathogen Salmonella enterica utilizes specific molecular mechanisms to attach to and colonize plants. This work characterizes two S. enterica gen...

  20. Bordetella pertussis isolates in Finland: Serotype and fimbrial expression

    PubMed Central

    Heikkinen, Eriikka; Xing, Dorothy K; Ölander, Rose-Marie; Hytönen, Jukka; Viljanen, Matti K; Mertsola, Jussi; He, Qiushui

    2008-01-01

    Background Bordetella pertussis causes whooping cough or pertussis in humans. It produces several virulence factors, of which the fimbriae are considered adhesins and elicit immune responses in the host. B. pertussis has three distinct serotypes Fim2, Fim3 or Fim2,3. Generally, B. pertussis Fim2 strains predominate in unvaccinated populations, whereas Fim3 strains are often isolated in vaccinated populations. In Finland, pertussis vaccination was introduced in 1952. The whole-cell vaccine contained two strains, 18530 (Fim3) since 1962 and strain 1772 (Fim2,3) added in 1976. After that the vaccine has remained the same until 2005 when the whole-cell vaccine was replaced by the acellular vaccine containing pertussis toxin and filamentous hemagglutinin. Our aims were to study serotypes of Finnish B. pertussis isolates from 1974 to 2006 in a population with > 90% vaccination coverage and fimbrial expression of the isolates during infection. Serotyping was done by agglutination and serotype-specific antibody responses were determined by blocking ELISA. Results Altogether, 1,109 isolates were serotyped. Before 1976, serotype distributions of Fim2, Fim3 and Fim2,3 were 67%, 19% and 10%, respectively. From 1976 to 1998, 94% of the isolates were Fim2 serotype. Since 1999, the frequency of Fim3 strains started to increase and reached 83% during a nationwide epidemic in 2003. A significant increase in level of serum IgG antibodies against purified fimbriae was observed between paired sera of 37 patients. The patients infected by Fim3 strains had antibodies which blocked the binding of monoclonal antibodies to Fim3 but not to Fim2. Moreover, about one third of the Fim2 strain infected patients developed antibodies capable of blocking of binding of both anti-Fim2 and Fim3 monoclonal antibodies. Conclusion Despite extensive vaccinations in Finland, B. pertussis Fim2 strains were the most common serotype. Emergence of Fim3 strains started in 1999 and coincided with nationwide

  1. Genomics Reveals the Worldwide Distribution of Multidrug-Resistant Serotype 6E Pneumococci

    PubMed Central

    van Tonder, Andries J.; Bray, James E.; Roalfe, Lucy; White, Rebecca; Zancolli, Marta; Quirk, Sigríður J.; Haraldsson, Gunnsteinn; Jolley, Keith A.; Maiden, Martin C. J.; Bentley, Stephen D.; Haraldsson, Ásgeir; Erlendsdóttir, Helga; Kristinsson, Karl G.; Goldblatt, David

    2015-01-01

    The pneumococcus is a leading pathogen infecting children and adults. Safe, effective vaccines exist, and they work by inducing antibodies to the polysaccharide capsule (unique for each serotype) that surrounds the cell; however, current vaccines are limited by the fact that only a few of the nearly 100 antigenically distinct serotypes are included in the formulations. Within the serotypes, serogroup 6 pneumococci are a frequent cause of serious disease and common colonizers of the nasopharynx in children. Serotype 6E was first reported in 2004 but was thought to be rare; however, we and others have detected serotype 6E among recent pneumococcal collections. Therefore, we analyzed a diverse data set of ∼1,000 serogroup 6 genomes, assessed the prevalence and distribution of serotype 6E, analyzed the genetic diversity among serogroup 6 pneumococci, and investigated whether pneumococcal conjugate vaccine-induced serotype 6A and 6B antibodies mediate the killing of serotype 6E pneumococci. We found that 43% of all genomes were of serotype 6E, and they were recovered worldwide from healthy children and patients of all ages with pneumococcal disease. Four genetic lineages, three of which were multidrug resistant, described ∼90% of the serotype 6E pneumococci. Serological assays demonstrated that vaccine-induced serotype 6B antibodies were able to elicit killing of serotype 6E pneumococci. We also revealed three major genetic clusters of serotype 6A capsular sequences, discovered a new hybrid 6C/6E serotype, and identified 44 examples of serotype switching. Therefore, while vaccines appear to offer protection against serotype 6E, genetic variants may reduce vaccine efficacy in the longer term because of the emergence of serotypes that can evade vaccine-induced immunity. PMID:25972423

  2. Genomics Reveals the Worldwide Distribution of Multidrug-Resistant Serotype 6E Pneumococci.

    PubMed

    van Tonder, Andries J; Bray, James E; Roalfe, Lucy; White, Rebecca; Zancolli, Marta; Quirk, Sigríður J; Haraldsson, Gunnsteinn; Jolley, Keith A; Maiden, Martin C J; Bentley, Stephen D; Haraldsson, Ásgeir; Erlendsdóttir, Helga; Kristinsson, Karl G; Goldblatt, David; Brueggemann, Angela B

    2015-07-01

    The pneumococcus is a leading pathogen infecting children and adults. Safe, effective vaccines exist, and they work by inducing antibodies to the polysaccharide capsule (unique for each serotype) that surrounds the cell; however, current vaccines are limited by the fact that only a few of the nearly 100 antigenically distinct serotypes are included in the formulations. Within the serotypes, serogroup 6 pneumococci are a frequent cause of serious disease and common colonizers of the nasopharynx in children. Serotype 6E was first reported in 2004 but was thought to be rare; however, we and others have detected serotype 6E among recent pneumococcal collections. Therefore, we analyzed a diverse data set of ∼1,000 serogroup 6 genomes, assessed the prevalence and distribution of serotype 6E, analyzed the genetic diversity among serogroup 6 pneumococci, and investigated whether pneumococcal conjugate vaccine-induced serotype 6A and 6B antibodies mediate the killing of serotype 6E pneumococci. We found that 43% of all genomes were of serotype 6E, and they were recovered worldwide from healthy children and patients of all ages with pneumococcal disease. Four genetic lineages, three of which were multidrug resistant, described ∼90% of the serotype 6E pneumococci. Serological assays demonstrated that vaccine-induced serotype 6B antibodies were able to elicit killing of serotype 6E pneumococci. We also revealed three major genetic clusters of serotype 6A capsular sequences, discovered a new hybrid 6C/6E serotype, and identified 44 examples of serotype switching. Therefore, while vaccines appear to offer protection against serotype 6E, genetic variants may reduce vaccine efficacy in the longer term because of the emergence of serotypes that can evade vaccine-induced immunity. PMID:25972423

  3. Prevalence of G and P serotypes among equine rotaviruses in the faeces of diarrhoeic foals.

    PubMed

    Browning, G F; Begg, A P

    1996-01-01

    Variant types of VP4 and VP7 gene segments of faecal rotaviruses from diarrhoeic foals were identified by restriction endonuclease digestion of reverse transcription/polymerase chain reaction (RT/PCR) products. The variants observed were correlated with serotypes by determination of the sequence of representative RT/PCR products (entire coding sequence for VP7 and the VP8 region of VP4) and comparison to published sequences of equine G and P serotype genes. Both G and P serotypes could be predicted for 95/116 (82%) strains, P serotype only for a further 8 (7%) strains and G serotype only for 1 (1%) strain. All characterised strains belonged to the same P serotype, P12, although minor sequence variations were observed. Of those strains able to be assigned to G serotypes, 84/96 (87.5%) belonged to serotype G3A, and 12/96 (12.5%) belonged to serotype G14. Comparison of G serotyping by ELISA to the RT/PCR method showed that serotyping equine rotaviruses by currently available ELISA methods was prone to error. This study establishes the restricted serotypic diversity of equine rotaviruses, and the significance of serotype G14. PMID:8712925

  4. Clinical differences among PCR-proven dengue serotype infections.

    PubMed

    Limkittikul, Kriengsak; Yingsakmongkon, Sangchai; Jittmittraphap, Akanitt; Chuananon, Somchai; Kongphrai, Yuphin; Kowasupathr, Surasak; Rojanawatsirivit, Chaiyaporn; Mammen, Mammen P; Jampangern, Wipawee

    2005-11-01

    The objective of this study was to compare the clinical spectra of the dengue serotypes proven by the PCR technique. This retrospective study reviewed the clinical information of dengue-infected patients who were admitted to northeastern provincial hospitals in Thailand from June to September 2002. Dengue infection and viral serotypes were confirmed by polymerase chain reaction (PCR). Paired anti-dengue immunoglobulin G (IgG) and IgM from paired sera were analyzed by enzyme-linked immunosorbent assay (ELISA). Ninety-nine PCR-proven dengue-infected Thai patients were studied. Their ages ranged from 3-30 years. They were infected with DEN1, DEN2, DEN3 and DEN4 in 21, 55, 12, and 12%, respectively. Twenty-two percent had primary and 78% had secondary infections. Dengue fever was the most common presentation for both primary (77.2%) and secondary infections (46.7%). The ratios of dengue fever:dengue hemorrhagic fever (DF:DHF) and non-dengue shock syndrome:dengue shock syndrome (non-DSS:DSS) for DEN2 was the lowest of the dengue serotypes. There was no difference in the duration of fever, percentage of hepatomegaly and bleeding among the serotypes in both DF and DHF. The trends in the white blood cells, lymphocyte and atypical lymphocyte counts in DEN3 were the highest, while those of DEN1 were the lowest of the dengue serotypes. PMID:16610645

  5. Botulinum neurotoxin serotypes detected by electrochemical impedance spectroscopy.

    PubMed

    Savage, Alison C; Buckley, Nicholas; Halliwell, Jennifer; Gwenin, Christopher

    2015-05-01

    Botulinum neurotoxin is one of the deadliest biological toxins known to mankind and is able to cause the debilitating disease botulism. The rapid detection of the different serotypes of botulinum neurotoxin is essential for both diagnosis of botulism and identifying the presence of toxin in potential cases of terrorism and food contamination. The modes of action of botulinum neurotoxins are well-established in literature and differ for each serotype. The toxins are known to specifically cleave portions of the SNARE proteins SNAP-25 or VAMP; an interaction that can be monitored by electrochemical impedance spectroscopy. This study presents a SNAP-25 and a VAMP biosensors for detecting the activity of five botulinum neurotoxin serotypes (A-E) using electrochemical impedance spectroscopy. The biosensors are able to detect concentrations of toxins as low as 25 fg/mL, in a short time-frame compared with the current standard methods of detection. Both biosensors show greater specificity for their compatible serotypes compared with incompatible serotypes and denatured toxins. PMID:25954998

  6. Botulinum Neurotoxin Serotypes Detected by Electrochemical Impedance Spectroscopy

    PubMed Central

    Savage, Alison C.; Buckley, Nicholas; Halliwell, Jennifer; Gwenin, Christopher

    2015-01-01

    Botulinum neurotoxin is one of the deadliest biological toxins known to mankind and is able to cause the debilitating disease botulism. The rapid detection of the different serotypes of botulinum neurotoxin is essential for both diagnosis of botulism and identifying the presence of toxin in potential cases of terrorism and food contamination. The modes of action of botulinum neurotoxins are well-established in literature and differ for each serotype. The toxins are known to specifically cleave portions of the SNARE proteins SNAP-25 or VAMP; an interaction that can be monitored by electrochemical impedance spectroscopy. This study presents a SNAP-25 and a VAMP biosensors for detecting the activity of five botulinum neurotoxin serotypes (A–E) using electrochemical impedance spectroscopy. The biosensors are able to detect concentrations of toxins as low as 25 fg/mL, in a short time-frame compared with the current standard methods of detection. Both biosensors show greater specificity for their compatible serotypes compared with incompatible serotypes and denatured toxins. PMID:25954998

  7. Inactivation of Salmonella enterica and Listeria monocytogenes in cantaloupe puree by high hydrostatic pressure with/without added ascorbic acid.

    PubMed

    Mukhopadhyay, Sudarsan; Sokorai, Kimberly; Ukuku, Dike; Fan, Xuetong; Juneja, Vijay; Sites, Joseph; Cassidy, Jennifer

    2016-10-17

    The objective of this research was to evaluate and develop a method for inactivation of Salmonella enterica and Listeria monocytogenes in cantaloupe puree (CP) by high hydrostatic pressure (HHP). Cantaloupe being the most netted varieties of melons presents a greater risk of pathogen transmission. Freshly prepared CP with or without 0.1% ascorbic acid (AA) was inoculated with a bacterial cocktail composed of a three serotype mixture of S. enterica (S. Poona, S. Newport H1275 and S. Stanley H0558) and a mixture of three strains of L. monocytogenes (Scott A, 43256 and 51742) to a population of ca. 10(8)CFU/g. Double sealed and double bagged inoculated CP (ca. 5g) were pressure treated at 300, 400 and 500MPa at 8°C and 15°C for 5min. Data indicated increased inactivation of both Salmonella and Listeria spp. with higher pressure. Log reduction for CP at 300MPa, 8°C for 5min was 2.4±0.2 and 1.6±0.5logCFU/g for Salmonella and Listeria, respectively. Survivability of the pathogens was significantly compromised at 400MPa and 8°C, inactivating 4.5±0.3logCFU/g of Salmonella and 3.0±0.4logCFU/g of Listeria spp. Complete inactivation of the pathogens in the puree (log reduction >6.7logCFU/g), with or without AA, was achieved when the pressure was further increased to 500MPa, except that for Listeria containing no AA at 8°C. Listeria presented higher resistance to pressure treatment compared to Salmonella spp. Initial temperatures (8 and 15°C) had no significant influence on Salmonella log reductions. Log reduction of pathogens increased but not significantly with increase of temperature. AA did not show any significant antimicrobial activity. Viable counts were about 0.2-0.4logCFU/g less in presence of 0.1% AA. These data validate that HHP can be used as an effective method for decontamination of cantaloupe puree. PMID:27441819

  8. Assessment of the effect of a Salmonella enterica ser. Typhimurium culture supernatant on the single-cell lag time of foodborne pathogens.

    PubMed

    Blana, Vasiliki A; Lianou, Alexandra; Nychas, George-John E

    2015-12-23

    The objective of this study was the in vitro evaluation of the effect of a cell-free microbial supernatant, produced by a luxS-positive Salmonella enterica ser. Typhimurium strain, on the single-cell growth kinetic behavior of two strains of S. enterica (serotypes Enteritidis and Typhimurium) and a methicillin-resistant Staphylococcus aureus strain. The single-cell lag time (λ) of the pathogens was estimated in the absence and presence (20% v/v) of microbial supernatant based on optical density measurements. As demonstrated by the obtained results, the tested microbial supernatant had a strain-specific effect on the single-cell λ and its variability. Although the mean λ values were similar in the absence and presence of microbial supernatant in the case of Salmonella Enteritidis, a significant (P ≤ 0.05) reduction and increase in the mean value of this parameter in the presence of microbial supernatant were observed for Salmonella Typhimurium and St. aureus, respectively. With regard to the effect of the tested microbial supernatant on the single-cell variability of λ, similar λ distributions were obtained in its absence and presence for S. Enteritidis, while considerable differences were noted for the other two tested organisms; the coefficient of variation of λ in the absence and presence of microbial supernatant was 41.6 and 69.8% for S. Typhimurium, respectively, with the corresponding values for St. aureus being 74.0 and 56.9%. As demonstrated by the results of bioassays, the tested microbial supernatant exhibited autoinducer-2 activity, indicating a potential association of such quorum sensing compounds with the observed effects. Although preliminary in nature, the collected data provide a good basis for future research on the role of quorum sensing in the single-cell growth behavior of foodborne pathogens. PMID:26433459

  9. Multidrug-resistant Salmonella enterica Serovar Typhimurium Monophasic Variant 4,12:i:- Isolated from Asymptomatic Wildlife in a Catalonian Wildlife Rehabilitation Center, Spain.

    PubMed

    Molina-López, Rafael A; Vidal, Anna; Obón, Elena; Martín, Marga; Darwich, Laila

    2015-07-01

    Wildlife can act as long-term asymptomatic reservoirs for zoonotic bacteria, such as Salmonella. The prevalence and antimicrobial-susceptibility profiles of Salmonella spp. were assessed in 263 cases in wildlife from 22 animal orders from a wildlife rehabilitation center in Catalonia (NE Spain), September 2013-May 2014. Eleven of 263 tested animals were positive for Salmonella spp., representing an overall prevalence of 4.2%. Prevalences by taxonomic categories were 2% in mammals, 4.7% in birds, and 4.5% in reptiles. By species, one each of European hedgehog (Erinaceus europeus; from a sample of n = 26), Eurasian Eagle Owl (Bubo bubo; n = 2), Barn Owl (Tyto alba; n = 3), Tawny Owl (Strix aluco; n = 20), Egyptian Vulture (Neophron percnopterus; n = 1), Griffon Vulture (Gyps fulvus; n = 1), and Hoopoe (Upupa epops; n = 2), and two each Common Kestrels (Falco tinnunculus; n = 16) and pond sliders (Trachemys scripta; n = 25) were positive for Salmonella. By serotyping, seven of eleven isolates were classified as S. enterica subsp. enterica serovar Typhimurium, and five of seven belonged to the monophasic variant 4,12:i:-. All the monophasic variants were isolated from birds (4/5 in raptors) and showed a multidrug-resistance (MDR) profile to at least ampicillin, streptomycin, sulfonamide, and tetracycline (R-type ASSuT), and up to 12 antibiotics. The large proportion of S. Typhimurium monophasic MDR strains detected in wildlife never treated with antibiotics, especially in raptors, adds more complexity to the epidemiologic control of one of the most frequent serovars involved in human and livestock infection. PMID:25973627

  10. Emergence and Diversity of Salmonella enterica Serovar Indiana Isolates with Concurrent Resistance to Ciprofloxacin and Cefotaxime from Patients and Food-Producing Animals in China.

    PubMed

    Bai, Li; Zhao, Jiayong; Gan, Xin; Wang, Juan; Zhang, Xiuli; Cui, Shenghui; Xia, Shengli; Hu, Yujie; Yan, Shaofei; Wang, Jiahui; Li, Fengqin; Fanning, Séamus; Xu, Jin

    2016-06-01

    Salmonellosis is a major global foodborne infection, and strains that are resistant to a great variety of antibiotics have become a major public health concern. The aim of this study was to identify genes conferring resistance to fluoroquinolones and extended-spectrum β-lactams in nontyphoidal Salmonella (NTS) from patients and food-producing animals in China. In total, 133 and 21 NTS isolates from animals and humans, respectively, exhibiting concurrent resistance to ciprofloxacin and cefotaxime were cultured independently from 2009 to ∼2013. All of the isolates were identified, serotyped, and subjected to antimicrobial susceptibility testing. Importantly, the isolates with concurrent resistance to ciprofloxacin and cefotaxime all were confirmed as S. enterica serovar Indiana. The presence of fluoroquinolone resistance genes and extended-spectrum β-lactamases (ESBLs) was established by PCR and DNA sequencing. The occurrence and diversity of different genes conferring fluoroquinolone resistance [qepA, oqxAB, and aac(6')-Ib-cr] with mutations in topoisomerase-encoding genes (gyrA and parC) and several ESBLs (including CTX-M-65, CTX-M-27, CTX-M-15, CTX-M-14, and CTX-M-14/CTX-M-15) were noteworthy. Genes located on mobile genetic elements were identified by conjugation and transformation. Pulsed-field gel electrophoresis, used to determine the genetic relationships between these isolates, generated 91 pulsotypes from 133 chicken isolates and 17 pulsotypes from the 21 clinical isolates that showed considerable diversity. Analysis of the pulsotypes obtained with the isolates showed some clones appeared to have existed for several years and had been disseminating between humans and food-producing animals. This study highlights the emergence of ciprofloxacin- and cefotaxime-resistant S. enterica serovar Indiana, posing a threat to public health. PMID:27001808

  11. Complete Genome Sequence of Salmonella enterica subsp. enterica Serovar Indiana C629, a Carbapenem-Resistant Bacterium Isolated from Chicken Carcass in China

    PubMed Central

    Liu, Feng; Peng, Zixin; Li, Fengqin

    2016-01-01

    The carbapenem-resistant Salmonella enterica subsp. enterica serovar Indiana strain C629 was isolated from a chicken carcass collected from a slaughterhouse in Qingdao, China. The complete genome sequence of C629 contains a circular 4,791,723-bp chromosome and a circular 210,106-bp plasmid. Genes involved in carbapenem resistance of this bacterium were identified by whole-genome analysis. PMID:27417837

  12. FDA Approves First Botulism Antitoxin for Use in Neutralizing All Seven Known Botulinum Nerve Toxin Serotypes

    MedlinePlus

    ... approves first Botulism Antitoxin for use in neutralizing all seven known botulinum nerve toxin serotypes Product to ... contains a mixture of antibody fragments that neutralize all of the seven botulinum nerve toxin serotypes known ...

  13. Development of a Pefloxacin Disk Diffusion Method for Detection of Fluoroquinolone-Resistant Salmonella enterica.

    PubMed

    Skov, Robert; Matuschek, Erika; Sjölund-Karlsson, Maria; Åhman, Jenny; Petersen, Andreas; Stegger, Marc; Torpdahl, Mia; Kahlmeter, Gunnar

    2015-11-01

    Fluoroquinolones (FQs) are among the drugs of choice for treatment of Salmonella infections. However, fluoroquinolone resistance is increasing in Salmonella due to chromosomal mutations in the quinolone resistance-determining regions (QRDRs) of the topoisomerase genes gyrA, gyrB, parC, and parE and/or plasmid-mediated quinolone resistance (PMQR) mechanisms including qnr variants, aac(6')-Ib-cr, qepA, and oqxAB. Some of these mutations cause only subtle increases in the MIC, i.e., MICs ranging from 0.12 to 0.25 mg/liter for ciprofloxacin (just above the wild-type MIC of ≤0.06 mg/liter). These isolates are difficult to detect with standard ciprofloxacin disk diffusion, and plasmid-mediated resistance, such as qnr, is often not detected by the nalidixic acid screen test. We evaluated 16 quinolone/fluoroquinolone disks for their ability to detect low-level-resistant Salmonella enterica isolates that are not serotype Typhi. A total of 153 Salmonella isolates characterized for the presence (n = 104) or absence (n = 49) of gyrA and/or parC topoisomerase mutations, qnrA, qnrB, qnrD, qnrS, aac(6')-Ib-cr, or qepA genes were investigated. All isolates were MIC tested by broth microdilution against ciprofloxacin, levofloxacin, and ofloxacin and by disk diffusion using EUCAST or CLSI methodology. MIC determination correctly categorized all isolates as either wild-type isolates (MIC of ≤0.06 mg/liter and absence of resistance genes) or non-wild-type isolates (MIC of >0.06 mg/liter and presence of a resistance gene). Disk diffusion using these antibiotics and nalidixic acid failed to detect some low-level-resistant isolates, whereas the 5-μg pefloxacin disk correctly identified all resistant isolates. However, pefloxacin will not detect isolates having aac(6')-Ib-cr as the only resistance determinant. The pefloxacin disk assay was approved and implemented by EUCAST (in 2014) and CLSI (in 2015). PMID:26292292

  14. Biofilms promote survival and virulence of Salmonella enterica sv. Tennessee during prolonged dry storage and after passage through an in vitro digestion system.

    PubMed

    Aviles, Bryan; Klotz, Courtney; Eifert, Joseph; Williams, Robert; Ponder, Monica

    2013-04-01

    Salmonella enterica serotypes have been linked to outbreaks associated with low water activity foods. While the biofilm-forming abilities of Salmonella improve its survival during thermal processing and sanitation it is unclear whether biofilms enhance survival to desiccation and gastric stresses. The purpose of this study was to quantify the effect of physiological state (planktonic versus biofilm) and prior exposure to desiccation and storage in dry milk powder on Salmonella survival and gene expression after passage through an in vitro digestion model. Planktonic cells of Salmonella enterica serotype Tennessee were deposited onto membranes while biofilms were formed on glass beads. The cells were subsequently dried at room temperature and stored in dried milk powder (a(w)=0.3) for up to 30 days. Salmonella survival was quantified by serial dilution onto Brilliant Green Agar before desiccation, after desiccation, after 1-day storage and after 30-day storage. At each sampling period both physiological states were tested for survival through a simulated gastrointestinal system. RNA was extracted at the identical time points and Quantitative Real-Time PCR was used to determine relative expression for genes associated with stress response (rpoS, otsB), virulence (hilA, invA, sipC) and a housekeeping gene 16S rRNA. The physiological state and length of storage affected the survival and gene expression of Salmonella within the desiccated milk powder environment and after passage through an in vitro digestion system (p<0.05). Larger numbers of S. Tennessee were recovered by plate counts for biofilms compared to planktonic, however, the numbers of Salmonella genomes detected by qPCR were not significantly different suggesting entry of the planktonic cells of S. Tennessee into a viable but non-culturable state. The increased expression of stress response genes rpoS and otsB correlated with survival, indicating cross-protection to low water activity and acid stress

  15. Human Adenovirus Serotype 3 Vector Packaged by a Rare Serotype 14 Hexon

    PubMed Central

    Ma, Qiang; Liu, Qian; Lu, Xiaomei; Zhou, Rong

    2016-01-01

    Recombinant adenovirus serotype 3 (rAd3), which infects cells through the receptor desmoglein 2 (DSG2), has been investigated as a vector for gene therapy or vaccination. However, pre-existing anti-vector immunity may limit the practical application of rAd3. In this study, we investigated the seroprevalence and neutralizing antibody (NAb) titers to Ad3 and alternate serotypes in normal healthy adults in southern China. Sera samples had a high seroprevalence (80.00%) against Ad3 and Ad7 (85.83%), compared with Ad14 (22.50%). Furthermore, 19.17% and 25.83% of samples had high-titer neutralizing antibodies to Ad3 and Ad7, respectively, compared with 3.33% against Ad14. We constructed a chimeric adenovirus, rAd3H14, designed to evade anti-vector immunity by replacing the enhanced green fluorescent protein (EGFP)-expressing hexon of the rAd3EGFP vector with a hexon from Ad14. The chimeric vector rAd3H14 was not neutralized in vitro efficiently by Ad3 NAbs using sera from mice and normal healthy human volunteers. Furthermore, in contrast to the unmodified vector rAd3EGFP, rAd3H14 induced robust antibody responses against EGFP in mice with high levels of pre-existing anti-Ad3 immunity. In conclusion, the chimeric vector rAd3H14 may be a useful alternative vector in adult populations with a high prevalence of Ad3 NAbs. PMID:27328032

  16. First detection of bluetongue virus serotype 14 in Poland.

    PubMed

    Orłowska, Anna; Trębas, Paweł; Smreczak, Marcin; Marzec, Anna; Żmudziński, Jan F

    2016-07-01

    Here, we present the first detected cases of bluetongue virus (BTV) in native cattle from Poland. The virus was found in animals located near the Polish-Belarusian and Polish-Lithuanian borders. The positive animals were detected through an official epidemiological surveillance program. A combination of type-specific real-time RT-PCR and phylogenetic tests revealed the presence of BTV serotype 14 (BTV-14). This serotype is highly homologous to the vaccine strain and BTV-14 present in Russia, Lithuania, and Spain (from an animal imported from Lithuania). The most probable route of virus introduction to Poland was transmission through midges. All of the cases were subclinical. PMID:27068167

  17. Comparison of Toxicological Properties of Botulinum Neurotoxin Serotypes A and B in Mice

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Botulinum neurotoxins (BoNTs) are among the most toxic biological toxins for humans. Of the seven known serotypes (A-G) of BoNT, serotypes A, B and E cause most of the human foodborne intoxications. In this study, we compared the toxicological properties of BoNT serotype A and B holotoxins and compl...

  18. Serotyping of Cryptococcus neoformans Isolates from Clinical and Environmental Sources in Spain

    PubMed Central

    Baró, Teresa; Torres-Rodríguez, Josep M.; Morera, Yolanda; Alía, Concepción; López, Olga; Méndez, Raul

    1999-01-01

    We determined biovars and serotypes of 154 isolates of Cryptococcus neoformans from clinical and environmental sources from different areas of Spain. All clinical isolates belonged to C. neoformans var. neoformans. Serotypes showed an irregular distribution. C. neoformans var. gattii serotype B was isolated from necropsy specimens from goats with pulmonary disease. PMID:10074545

  19. Comparison of a Real-Time Multiplex PCR and Sequetyping Assay for Pneumococcal Serotyping

    PubMed Central

    Robberts, Lourens; Wolter, Nicole; Nicol, Paul; Mafofo, Joseph; Africa, Samantha; Zar, Heather J.; Nicol, Mark P.

    2015-01-01

    Background Pneumococcal serotype identification is essential to monitor pneumococcal vaccine effectiveness and serotype replacement. Serotyping by conventional serological methods are costly, labour-intensive, and require significant technical expertise. We compared two different molecular methods to serotype pneumococci isolated from the nasopharynx of South African infants participating in a birth cohort study, the Drakenstein Child Health Study, in an area with high 13-valent pneumococcal conjugate vaccine (PCV13) coverage. Methods A real-time multiplex PCR (rmPCR) assay detecting 21 different serotypes/-groups and a sequetyping assay, based on the sequence of the wzh gene within the pneumococcal capsular locus, were compared. Forty pneumococcal control isolates, with serotypes determined by the Quellung reaction, were tested. In addition, 135 pneumococcal isolates obtained from the nasopharynx of healthy children were tested by both serotyping assays and confirmed by Quellung testing. Discordant results were further investigated by whole genome sequencing of four isolates. Results Of the 40 control isolates tested, 25 had a serotype covered by the rmPCR assay. These were all correctly serotyped/-grouped. Sequetyping PCR failed in 7/40 (18%) isolates. For the remaining isolates, sequetyping assigned the correct serotype/-group to 29/33 (88%) control isolates. Of the 132/135 (98%) nasopharyngeal pneumococcal isolates that could be typed, 69/132 (52%) and 112/132 (85%) were assigned the correct serotype/-group by rmPCR and sequetyping respectively. The serotypes of 63/132 (48%) isolates were not included in the rmPCR panel. All except three isolates (serotype 25A and 38) were theoretically amplified and differentiated into the correct serotype/-group with some strains giving ambigous results (serotype 13/20, 17F/33C, and 11A/D/1818F). Of the pneumococcal serotypes detected in this study, 69/91 (76%) were not included in the current PCV13. The most frequently

  20. Purification and characterization of serotype 6 fimbriae from Bordetella pertussis and comparison of their properties with serotype 2 fimbriae.

    PubMed

    Cowell, J L; Zhang, J M; Urisu, A; Suzuki, A; Steven, A C; Liu, T; Liu, T Y; Manclark, C R

    1987-04-01

    Fimbriae were removed from Bordetella pertussis (serotype 1.3.6) by mechanical shearing and purified by precipitation with ammonium sulfate, pH-dependent precipitation at pH 7.4, followed by two successive extractions of the precipitated fimbriae with 4 M urea. By electron microscopy, the precipitated fimbriae appeared as aggregated bundles of long, relatively straight filaments which were disaggregated to individual flexuous filaments at pH 10.5. These purified fimbriae were identified as serotype 6 agglutinogens, since antibody to the purified fimbriae agglutinated B. pertussis strains serotyped as 1.3.6, 1.2.3.6, or 1.2.3.4.6 but did not agglutinate strains of serotype 1.2.3.4, 1.2.3, or 1.3. In contrast, antibody to serotype 2 fimbriae only agglutinated B. pertussis strains containing serotype 2 agglutinogen. Purified type 6 and 2 fimbriae were found to be weakly cross-reactive by enzyme-linked immunosorbent assay, using polyclonal antibody to each type of fimbria. In an immunoblot assay, polyclonal antibodies to a 22,000-dalton subunit of fimbriae from B. bronchiseptica reacted strongly with the type 2 fimbrial subunit of B. pertussis, but only weakly with the type 6 subunit. When subjected to sodium dodecyl sulfate-polyacrylamide gel electrophoresis, the protein subunit of the type 6 fimbriae migrated with a molecular weight of 21,500, whereas the type 2 fimbrial subunit had a molecular weight of 22,000. The two types of subunits had similar amino acid compositions and showed amino-terminal sequence homology in 15 of 21 amino acids. The amino-terminal amino acid sequences of the B. pertussis fimbriae were distinct from those reported for fimbriae from other gram-negative bacteria. Neither the type 6 nor the type 2 fimbriae caused hemagglutination when assayed with several types of erythrocytes. PMID:2881893

  1. Purification and characterization of serotype 6 fimbriae from Bordetella pertussis and comparison of their properties with serotype 2 fimbriae.

    PubMed Central

    Cowell, J L; Zhang, J M; Urisu, A; Suzuki, A; Steven, A C; Liu, T; Liu, T Y; Manclark, C R

    1987-01-01

    Fimbriae were removed from Bordetella pertussis (serotype 1.3.6) by mechanical shearing and purified by precipitation with ammonium sulfate, pH-dependent precipitation at pH 7.4, followed by two successive extractions of the precipitated fimbriae with 4 M urea. By electron microscopy, the precipitated fimbriae appeared as aggregated bundles of long, relatively straight filaments which were disaggregated to individual flexuous filaments at pH 10.5. These purified fimbriae were identified as serotype 6 agglutinogens, since antibody to the purified fimbriae agglutinated B. pertussis strains serotyped as 1.3.6, 1.2.3.6, or 1.2.3.4.6 but did not agglutinate strains of serotype 1.2.3.4, 1.2.3, or 1.3. In contrast, antibody to serotype 2 fimbriae only agglutinated B. pertussis strains containing serotype 2 agglutinogen. Purified type 6 and 2 fimbriae were found to be weakly cross-reactive by enzyme-linked immunosorbent assay, using polyclonal antibody to each type of fimbria. In an immunoblot assay, polyclonal antibodies to a 22,000-dalton subunit of fimbriae from B. bronchiseptica reacted strongly with the type 2 fimbrial subunit of B. pertussis, but only weakly with the type 6 subunit. When subjected to sodium dodecyl sulfate-polyacrylamide gel electrophoresis, the protein subunit of the type 6 fimbriae migrated with a molecular weight of 21,500, whereas the type 2 fimbrial subunit had a molecular weight of 22,000. The two types of subunits had similar amino acid compositions and showed amino-terminal sequence homology in 15 of 21 amino acids. The amino-terminal amino acid sequences of the B. pertussis fimbriae were distinct from those reported for fimbriae from other gram-negative bacteria. Neither the type 6 nor the type 2 fimbriae caused hemagglutination when assayed with several types of erythrocytes. Images PMID:2881893

  2. Changes in the Porcine Intestinal Microbiome in Response to Infection with Salmonella enterica and Lawsonia intracellularis

    PubMed Central

    Singer, Randall S.; Gebhart, Connie J.; Sreevatsan, Srinand; Johnson, Timothy; Isaacson, Richard E.

    2015-01-01

    Salmonella enterica is a leading cause of food borne illness. Recent studies have shown that S. enterica is a pathogen capable of causing alterations to the composition of the intestinal microbiome. A recent prospective study of French pork production farms found a statistically significant association between Lawsonia intracellularis and carriage of S. enterica. In the current study the composition of the gut microbiome was determined in pigs challenged with S. enterica serovar Typhimurium and or L. intracellularis and compared to non-challenged control pigs. Principal coordinate analysis demonstrated that there was a disruption in the composition of the gut microbiome in the colon and cecum of pigs challenged with either pathogen. The compositions of the microbiomes of challenged pigs were similar to each other but differed from the non-challenged controls. There also were statistically significant increases in Anaerobacter, Barnesiella, Pediococcus, Sporacetigenium, Turicibacter, Catenibacterium, Prevotella, Pseudobutyrivibrio, and Xylanibacter in the challenged pigs. To determine if these changes were specific to experimentally challenged pigs, we determined the compositions of the fecal microbiomes of naturally infected pigs that were carriers of S. enterica. Pigs that were frequent shedders of S. enterica were shown to have similar fecal microbiomes compared to non-shedders or pigs that shed S. enterica infrequently. In a comparison of the differentially abundant bacteria in the naturally infected pigs compared to experimentally challenged pigs, 9 genera were differentially abundant and each exhibited the same increase or decrease in abundance between the two groups. Thus, there were similar changes in the GI microbiome associated with carriage of S. enterica regardless of whether the pigs were experimentally challenged with S. enterica or acquired it naturally. PMID:26461107

  3. Changes in the Porcine Intestinal Microbiome in Response to Infection with Salmonella enterica and Lawsonia intracellularis.

    PubMed

    Borewicz, Klaudyna A; Kim, Hyeun Bum; Singer, Randall S; Gebhart, Connie J; Sreevatsan, Srinand; Johnson, Timothy; Isaacson, Richard E

    2015-01-01

    Salmonella enterica is a leading cause of food borne illness. Recent studies have shown that S. enterica is a pathogen capable of causing alterations to the composition of the intestinal microbiome. A recent prospective study of French pork production farms found a statistically significant association between Lawsonia intracellularis and carriage of S. enterica. In the current study the composition of the gut microbiome was determined in pigs challenged with S. enterica serovar Typhimurium and or L. intracellularis and compared to non-challenged control pigs. Principal coordinate analysis demonstrated that there was a disruption in the composition of the gut microbiome in the colon and cecum of pigs challenged with either pathogen. The compositions of the microbiomes of challenged pigs were similar to each other but differed from the non-challenged controls. There also were statistically significant increases in Anaerobacter, Barnesiella, Pediococcus, Sporacetigenium, Turicibacter, Catenibacterium, Prevotella, Pseudobutyrivibrio, and Xylanibacter in the challenged pigs. To determine if these changes were specific to experimentally challenged pigs, we determined the compositions of the fecal microbiomes of naturally infected pigs that were carriers of S. enterica. Pigs that were frequent shedders of S. enterica were shown to have similar fecal microbiomes compared to non-shedders or pigs that shed S. enterica infrequently. In a comparison of the differentially abundant bacteria in the naturally infected pigs compared to experimentally challenged pigs, 9 genera were differentially abundant and each exhibited the same increase or decrease in abundance between the two groups. Thus, there were similar changes in the GI microbiome associated with carriage of S. enterica regardless of whether the pigs were experimentally challenged with S. enterica or acquired it naturally. PMID:26461107

  4. Molecular clonality and antimicrobial resistance in Salmonella enterica serovars Enteritidis and Infantis from broilers in three Northern regions of Iran

    PubMed Central

    2013-01-01

    Background Multidrug-resistant Salmonella strains are frequently encountered problems worldwide with considerable increased occurrences in recent years. The aim of this study was to investigate the occurrence and frequency of antimicrobial resistance and associated resistance genes in Salmonella isolates from broiler farms in different regions of Iran covering a time period of four years. Results From 2007 to 2011, 36 Salmonella strains were isolated from broiler farms located in three northern provinces of Iran. The isolates were serotyped, antimicrobial susceptibility tested, and characterized for antimicrobial resistance genes associated to the phenotype. Pulsed-field gel electrophoresis (PFGE) was applied for comparison of genetic relatedness. Two serovars were detected among the isolates; Salmonella enterica serovar Infantis (75%) and S. Enteritidis (25%). Thirty-four (94%) of the isolates exhibited resistance to nalidixic acid and ciprofloxacin caused by a single mutation in the quinolone resistance-determining region (QRDR) of gyrA. For all strains this mutation occurred in the codon of Asp87 leading to a Asp87-Tyr, Asp87-Gly or Asp87-Asn substitutions. All S. Infantis (n = 27) were resistant to tetracycline, spectinomycin, streptomycin, and sulfamethoxazole and harbored the associated resistance genes; tetA, dfrA14, aadA1, and sulI together with class 1 integrons. The isolates revealed highly similar PFGE patterns indicating clonal relatedness across different geographical locations. Conclusion The data provided fundamental information applicable when launching future control programs for broilers in Iran with the aim to conserve the effectiveness of important antimicrobials for treatment in humans. PMID:23561048

  5. Phenotypic and Genotypic Characterization of Salmonella enterica Recovered from Poultry Meat in Tunisia and Identification of New Genetic Traits

    PubMed Central

    Soufi, Leila; Sáenz, Yolanda; de Toro, María; Salah Abbassi, Mohamed; Rojo-Bezares, Beatriz; Vinué, Laura; Bouchami, Ons; Touati, Arabella; Ben Hassen, Assia; Hammami, Salah

    2012-01-01

    Abstract Thirty-seven Salmonella enterica isolates obtained from poultry meat in Tunisia were included in this study for characterization of antibiotic resistance mechanisms. High percentages of resistance were detected to ampicillin, sulfonamides, tetracycline, nalidixic acid, and streptomycin (32.4%–89.2%), and lower percentages to amoxicillin–clavulanic acid, kanamycin, amikacin, trimethoprim–sulfamethoxazol, and chloramphenicol (2.7%–18.9%). All strains showed susceptibility to ceftazidime, cefotaxime, gentamicin, and ciprofloxacin. Class 1 integrons were detected in 30% of Salmonella isolates, and four different gene cassette arrangements were detected, including genes implicated in resistance to aminoglycosides (aadA1 and aadA2) and trimethoprim (dfrA1). Four different Pc variants (PcW, PcH1, PcH1TTN-10, PcWTGN-10) with inactive P2 have been found among these isolates. Integron-positive isolates were ascribed to eight different serotypes. A Salmonella Schwarzengrund isolate harbored a new class 1 integron containing the qacH-dfrA1b-aadA1b-catB2 gene cassette arrangement, with the very unusual PcH1TTN-10 promoter, which has been registered in GenBank (accession no. HQ874651). Different plasmid replicon types were demonstrated among integron-positive isolates: IncI1 (8 isolates), IncN (8), IncP (2), IncFIB (2), and IncFII (2). Ten different pulsed-field gel electrophoresis profiles were detected among the 11 integron-positive isolates and 8 different sequence types were identified by multilocus sequence typing, one of them (registered as ST867) was new, detected in 3 Salmonella Zanzibar isolates. A high diversity of clones is observed among poultry Salmonella isolates and a high proportion of them show a multiresistant phenotype with very diverse mobile genetic structures that could be implicated in bacterial dissemination in different environments. PMID:21919733

  6. Phylogenetic Diversity of the Enteric Pathogen Salmonella enterica subsp. enterica Inferred from Genome-Wide Reference-Free SNP Characters

    PubMed Central

    Timme, Ruth E.; Pettengill, James B.; Allard, Marc W.; Strain, Errol; Barrangou, Rodolphe; Wehnes, Chris; Van Kessel, JoAnn S.; Karns, Jeffrey S.; Musser, Steven M.; Brown, Eric W.

    2013-01-01

    The enteric pathogen Salmonella enterica is one of the leading causes of foodborne illness in the world. The species is extremely diverse, containing more than 2,500 named serovars that are designated for their unique antigen characters and pathogenicity profiles—some are known to be virulent pathogens, while others are not. Questions regarding the evolution of pathogenicity, significance of antigen characters, diversity of clustered regularly interspaced short palindromic repeat (CRISPR) loci, among others, will remain elusive until a strong evolutionary framework is established. We present the first large-scale S. enterica subsp. enterica phylogeny inferred from a new reference-free k-mer approach of gathering single nucleotide polymorphisms (SNPs) from whole genomes. The phylogeny of 156 isolates representing 78 serovars (102 were newly sequenced) reveals two major lineages, each with many strongly supported sublineages. One of these lineages is the S. Typhi group; well nested within the phylogeny. Lineage-through-time analyses suggest there have been two instances of accelerated rates of diversification within the subspecies. We also found that antigen characters and CRISPR loci reveal different evolutionary patterns than that of the phylogeny, suggesting that a horizontal gene transfer or possibly a shared environmental acquisition might have influenced the present character distribution. Our study also shows the ability to extract reference-free SNPs from a large set of genomes and then to use these SNPs for phylogenetic reconstruction. This automated, annotation-free approach is an important step forward for bacterial disease tracking and in efficiently elucidating the evolutionary history of highly clonal organisms. PMID:24158624

  7. Evaluation of aroA deletion mutant of Salmonella enterica subspecies enterica serovar Abortusequi for its vaccine candidate potential.

    PubMed

    Alam, Javad; Singh, B R; Hansda, D; Singh, V P; Verma, J C

    2009-11-01

    The present study on a defined deletion aroA mutant (B-26) of Salmonella enterica subspecies enterica serovar Abortusequi (S. Abortusequi) for residual virulence and safety in experimental model revealed that the virulence of the strain was at no difference in any of the cell assays (caprine alveolar macrophages, bovine alveolar macrophages, guinea pig blood mononuclear cells and horse blood mononuclear cells) than that of its parent virulence plasmid cured (S-787) and wild type (E-156) strains. The mutant did not cause any apparent illness in baby guinea pigs (15 days old), adult male and female guinea pigs and also not in pregnant (54-55 days of gestation) guinea pigs through oral (4.2 x 10(9) cfu/ animal) and intramuscular (im) routes (4.2 x 10(7) cfu/ animal). In pregnant females the mutant also induced abortion as its parent (E-156) though to lesser extent (33%) than the parent strain (100%) on inoculation through intravaginal (4.2 x 10(9) cfu/ animal) and intraperitoneal (4.2 x 10(7) cfu/ animal) routes. The babies born from mutant inoculated mothers survived better and were also resistant to intraperitoneal lethal challenge (7.82 x 10(9) cfu/ animal) with 100% protection. Female guinea pigs challenged after 135-165 days of inoculation with the mutant afforded 100% protection from abortion and mortality caused by lethal infection (7.82 x 10(9) cfu/ animal) of wild type S. enterica Abortusequi (E-156). The study revealed that aroA mutant (B-26) was safe through oral and im routes for immunization and afforded 100% protection against salmonellosis for more than 5.5 months in guinea pigs. Although immunization with aroA mutant in experimental model afforded good protection against abortion and mortality induced by S. Abortusequi, further studies are needed in horses to exploit the strain's vaccine potential in the natural host. PMID:20099460

  8. PCR-Based Serotyping of Streptococcus pneumoniae from Culture-Negative Specimens: Novel Primers for Detection of Serotypes within Serogroup 18.

    PubMed

    Tanmoy, Arif M; Saha, Senjuti; Darmstadt, Gary L; Whitney, Cynthia G; Saha, Samir K

    2016-08-01

    Six multiplex-compatible PCR primers were designed to distinguish Streptococcus pneumoniae serotypes within serogroup 18 from culturable/nonculturable pneumococcal specimens, with no cross-reactivity with other serotypes and respiratory organisms. These primers will aid in the generation of better data on vaccine/nonvaccine serotypes in invasive and carriage pneumococcal surveillance and contribute to future vaccine formulation and impact studies. PMID:27252464

  9. PCR-Based Serotyping of Streptococcus pneumoniae from Culture-Negative Specimens: Novel Primers for Detection of Serotypes within Serogroup 18

    PubMed Central

    Tanmoy, Arif M.; Saha, Senjuti; Darmstadt, Gary L.; Whitney, Cynthia G.

    2016-01-01

    Six multiplex-compatible PCR primers were designed to distinguish Streptococcus pneumoniae serotypes within serogroup 18 from culturable/nonculturable pneumococcal specimens, with no cross-reactivity with other serotypes and respiratory organisms. These primers will aid in the generation of better data on vaccine/nonvaccine serotypes in invasive and carriage pneumococcal surveillance and contribute to future vaccine formulation and impact studies. PMID:27252464

  10. The Widespread Multidrug-Resistant Serotype O12 Pseudomonas aeruginosa Clone Emerged through Concomitant Horizontal Transfer of Serotype Antigen and Antibiotic Resistance Gene Clusters

    PubMed Central

    Thrane, Sandra Wingaard; Taylor, Véronique L.; Freschi, Luca; Kukavica-Ibrulj, Irena; Boyle, Brian; Laroche, Jérôme; Pirnay, Jean-Paul; Lévesque, Roger C.

    2015-01-01

    ABSTRACT The O-specific antigen (OSA) in Pseudomonas aeruginosa lipopolysaccharide is highly varied by sugar identity, side chains, and bond between O-repeats. These differences classified P. aeruginosa into 20 distinct serotypes. In the past few decades, O12 has emerged as the predominant serotype in clinical settings and outbreaks. These serotype O12 isolates exhibit high levels of resistance to various classes of antibiotics. Here, we explore how the P. aeruginosa OSA biosynthesis gene clusters evolve in the population by investigating the association between the phylogenetic relationships among 83 P. aeruginosa strains and their serotypes. While most serotypes were closely linked to the core genome phylogeny, we observed horizontal exchange of OSA biosynthesis genes among phylogenetically distinct P. aeruginosa strains. Specifically, we identified a “serotype island” ranging from 62 kb to 185 kb containing the P. aeruginosa O12 OSA gene cluster, an antibiotic resistance determinant (gyrAC248T), and other genes that have been transferred between P. aeruginosa strains with distinct core genome architectures. We showed that these genes were likely acquired from an O12 serotype strain that is closely related to P. aeruginosa PA7. Acquisition and recombination of the “serotype island” resulted in displacement of the native OSA gene cluster and expression of the O12 serotype in the recipients. Serotype switching by recombination has apparently occurred multiple times involving bacteria of various genomic backgrounds. In conclusion, serotype switching in combination with acquisition of an antibiotic resistance determinant most likely contributed to the dissemination of the O12 serotype in clinical settings. PMID:26396243

  11. Impact of pneumococcal vaccination in children on serotype distribution in adult community-acquired pneumonia using the serotype-specific multiplex urinary antigen detection assay.

    PubMed

    Pletz, Mathias W; Ewig, Santiago; Rohde, Gernot; Schuette, Hartwig; Rupp, Jan; Welte, Tobias; Suttorp, Norbert; Forstner, Christina

    2016-04-29

    The aim of the study was to compare the distribution of the vaccine-serotypes covered by pneumococcal conjugate vaccines (PCV7 and PCV13) in adult patients with pneumococcal community-acquired pneumonia in Germany between the periods 2002-2006 and 2007-2011 using a novel serotype-specific multiplex urinary antigen detection assay (SSUA). Vaccination of children started with PCV7 in 2007, which was replaced by PCV13 in 2010. Following confirmation of the accuracy of SSUA in long-term stored urine samples from 112 patients with confirmed pneumonia and known pneumococcal serotype, urine samples of 391 CAPNETZ patients with documented pneumococcal pneumonia (i.e. positive BinaxNOW(®) Streptococcus pneumoniae urine antigen test) but unknown serotype were tested for the 13 vaccine-serotypes using SSUA. The proportion of PCV7-serotypes significantly decreased in adult patients with pneumonia from 30.6% (2002-6) to 13.3% (2007-11, p<0.001); in bacteremic pneumonia, PCV7-serotypes completely disappeared (3/14 versus 0/19, p=0.058). Conversely, pneumococcal serotypes included by PCV13 remained stable during study period with a coverage of 61.5% (2002-06) and 59.7% (2007-11) in non-bacteremic pneumonia and 79% (for both periods) in bacteremic pneumonia, mainly due to an increase in pneumococcal serotypes 1, 3 and 7F during the second period. Thus, implementation of PCV7 in children in Germany in 2007 was associated with a significant decrease in vaccine-serotypes covered by PCV7 in adult patients with non-bacteremic pneumococcal pneumonia and with an elimination of PCV7 vaccine-serotypes in bacteremic pneumococcal pneumonia. PCV13 coverage remained high up to 2011, mainly due to an increase in serotypes 1, 3 and 7F. German Clinical Trials Register: DRKS00005274. PMID:27016653

  12. Selection of a screening panel of rhinoviral serotypes.

    PubMed

    Tomkinson, Nick; Wenlock, Mark; McCrae, Christopher

    2012-12-15

    A novel methodology for the selection of a representative primary and secondary screening panel of rhinoviral serotypes for the purposes of identifying potential antiviral agents is presented. This methodology focuses on the active-sites of the rhinoviral proteins but does not invoke historical SAR data, thereby avoiding compound bias. PMID:23122823

  13. Urban epidemic of dengue virus serotype 3 infection, Senegal, 2009.

    PubMed

    Faye, Ousmane; Ba, Yamar; Faye, Oumar; Talla, Cheikh; Diallo, Diawo; Chen, Rubing; Mondo, Mireille; Ba, Rouguiétou; Macondo, Edgard; Siby, Tidiane; Weaver, Scott C; Diallo, Mawlouth; Sall, Amadou Alpha

    2014-03-01

    An urban epidemic of dengue in Senegal during 2009 affected 196 persons and included 5 cases of dengue hemorrhagic fever and 1 fatal case of dengue shock syndrome. Dengue virus serotype 3 was identified from all patients, and Aedes aegypti mosquitoes were identified as the primary vector of the virus. PMID:24572297

  14. AAV Hybrid Serotypes: Improved Vectors for Gene Delivery

    PubMed Central

    Choi, Vivian W.; McCarty, Douglas M.; Samulski, R. Jude

    2006-01-01

    In recent years, significant efforts have been made on studying and engineering adeno-associated virus (AAV) capsid, in order to increase efficiency in targeting specific cell types that are non-permissive to wild type (wt) viruses and to improve efficacy in infecting only the cell type of interest. With our previous knowledge of the viral properties of the naturally occurring serotypes and the elucidation of their capsid structures, we can now generate capsid mutants, or hybrid serotypes, by various methods and strategies. In this review, we summarize the studies performed on AAV retargeting, and categorize the available hybrid serotypes to date, based on the type of modification: 1) transcapsidation, 2) adsorption of bi-specific antibody to capsid surface, 3) mosaic capsid, and 4) chimeric capsid. Not only these hybrid serotypes could achieve high efficiency of gene delivery to a specific targeted cell type, which can be better-tailored for a particular clinical application, but also serve as a tool for studying AAV biology such as receptor binding, trafficking and genome delivery into the nucleus. PMID:15975007

  15. Molecular serotyping of Escherichia coli: A verification and reclassification

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Background: Serotyping of E. coli, based on the O- (polysaccharide side chain) and H- (flagellar) antigens using antisera is a common practice for diagnostics, outbreak investigations, and epidemiological surveillance. The full set of E. coli serogroups comprises O-groups O1 to O181, with several O...

  16. Bluetongue Virus Serotype 8 Reemergence in Germany, 2007 and 2008

    PubMed Central

    Hoffmann, Bernd; Saßerath, Michael; Thalheim, Sabine; Bunzenthal, Claudia; Strebelow, Günter

    2008-01-01

    Reemerging bluetongue virus serotype 8 (BTV-8) in Germany was detected first in May 2007 in a sentinel cow and in February 2008 in an export heifer. Reemergence was confirmed by retesting the samples, experimental inoculation, fingerprinting analysis, and virus isolation. Overwintering of BTV-8 and continuous low-level infections are assumed. PMID:18760009

  17. Isolation of Vibrio cholerae serotype Ogawa from a Florida estuary.

    PubMed

    Motes, M L; Zywno, S R; DePaola, A; Becker, R E; Presnell, M W

    1983-01-01

    Vibrio cholerae serotype Ogawa was recently isolated from the estuarine waters of Apalachicola Bay, Fla., in areas that are subject to consistent fecal contamination and in areas that are remote from any apparent source of contamination. The significance of these organisms in the environment has not been determined. PMID:6824323

  18. Detection of foot-and-mouth disease serotype O by ELISA using a monoclonal antibody.

    PubMed

    Chen, Hao-Tai; Peng, Yun-Hua; Zhang, Yong-Guang; Liu, Xiang-Tao

    2013-02-01

    An ELISA assay with monoclonal antibody (MELISA) was used to type serotype O of foot-and-mouth disease virus (FMDV). All FMDV serotype O reference strains were positive by MELISA, while other viruses such as FMDV serotypes Asia 1, C, and A and classical swine fever virus, swine vesicular disease virus, and porcine reproductive and respiratory syndrome virus remained negative. Furthermore, FMDV serotype O positive samples were able to be detected by MELISA. This assay may be particularly suitable for diagnosis of FMDV serotype O infection in field stations. PMID:23600506

  19. Detection of foot-and-mouth disease serotype O by ELISA using a monoclonal antibody.

    PubMed

    Chen, Hao-tai; Peng, Yun-hua; Zhang, Yong-guang; Liu, Xiang-tao

    2012-12-01

    An ELISA assay with monoclonal antibody (MELISA) was used to type serotype O of foot-and-mouth disease virus (FMDV). All FMDV serotype O reference strains were positive by MELISA, while other viruses such as FMDV serotypes Asia 1, C, A and classical swine fever virus, swine vesicular disease virus, and porcine reproductive and respiratory syndrome virus remained negative. Further, FMDV serotype O positive samples were able to be detected by MELISA. This assay may be particularly suitable for diagnosis of FMDV serotype O infection in field stations. PMID:23244327

  20. Emergence of Bluetongue Virus Serotype 1 in French Corsica Island in September 2013.

    PubMed

    Sailleau, C; Viarouge, C; Bréard, E; Perrin, J B; Doceul, V; Vitour, D; Zientara, S

    2015-10-01

    Since 2000, French Corsica Island has been exposed to the emergence of three different BT virus (BTV) serotypes: serotype 2 in 2000 and 2001, serotype 4 in 2003 and serotype 16 in 2004. Between 2005 and August 2013, no outbreaks have been reported in the French Island. At the beginning of September 2013, sheep located in the south of the island showed clinical signs suggestive of BTV infection. Laboratory analyses identified the virus as BTV serotype 1. Phylogenetic studies showed that the sequences of this strain are closely related to the BTV-1 strain that was circulating in the Mediterranean basin and in Sardinia in 2012. PMID:24456375

  1. Viral agents associated with infantile gastroenteritis in Nigeria: relative prevalence of adenovirus serotypes 40 and 41, astrovirus, and rotavirus serotypes 1 to 4.

    PubMed

    Avery, R M; Shelton, A P; Beards, G M; Omotade, O O; Oyejide, O C; Olaleye, D O

    1992-06-01

    Sixty-six stool specimens from infants with diarrhoea in Nigeria were examined for the presence of viral pathogens. Rotaviruses were found in 25.8% of specimens and astroviruses in 1.5%. Serotypes were determined for 47.1% of the rotavirus positive specimens, all of which were serotype 1. RNA analysis revealed no unusual electrophoretic profiles. No enteric adenoviruses were detected. In contrast, in a parallel study conducted in the UK, rotaviruses (including serotypes 1, 2 and 4) accounted for 21.9% of infections, adenovirus serotypes 40 and 41 13.6%, and astroviruses 4.5%. PMID:1323591

  2. Group B streptococcus serotype prevalence in reproductive-age women at a tertiary care military medical center relative to global serotype distribution

    PubMed Central

    2010-01-01

    Background Group B Streptococcus (GBS) serotype (Ia, Ib, II-IX) correlates with pathogen virulence and clinical prognosis. Epidemiological studies of seroprevalence are an important metric for determining the proportion of serotypes in a given population. The purpose of this study was to evaluate the prevalence of individual GBS serotypes at Madigan Healthcare System (Madigan), the largest military tertiary healthcare facility in the Pacific Northwestern United States, and to compare seroprevalences with international locations. Methods To determine serotype distribution at Madigan, we obtained GBS isolates from standard-of-care anogenital swabs from 207 women of indeterminate gravidity between ages 18-40 during a five month interval. Serotype was determined using a recently described molecular method of polymerase chain reaction by capsular polysaccharide synthesis (cps) genes associated with pathogen virulence. Results Serotypes Ia, III, and V were the most prevalent (28%, 27%, and 17%, respectively). A systematic review of global GBS seroprevalence, meta-analysis, and statistical comparison revealed strikingly similar serodistibution at Madigan relative to civilian-sector populations in Canada and the United States. Serotype Ia was the only serotype consistently higher in North American populations relative to other geographic regions (p < 0.005). The number of non-typeable isolates was significantly lower in the study (p < 0.005). Conclusion This study establishes PCR-based serotyping as a viable strategy for GBS epidemiological surveillance. Our results suggest that GBS seroprevalence remains stable in North America over the past two decades. PMID:21106080

  3. Implementation of Salmonella serotype determination using pulsed-field gel electrophoresis in a state public health laboratory.

    PubMed

    Bopp, Dianna J; Baker, Deborah J; Thompson, Lisa; Saylors, Amy; Root, Timothy P; Armstrong, Leeanna; Mitchell, Kara; Dumas, Nellie B; Musser, Kimberlee Arruda

    2016-08-01

    We examined the use of pulsed-field gel electrophoresis (PFGE) to predict serotype for Salmonella isolates. Between 2012 and 2014 we assessed 4481 isolates, resulting in >90% assigned serotypes. PFGE is efficient for determining serotype in the majority of cases and results in expedited serotype determination, as well as cost savings. PMID:27220605

  4. Chlorhexidine susceptibilities of mutans streptococcal serotypes and ribotypes.

    PubMed Central

    Grönroos, L; Mättö, J; Saarela, M; Luoma, A R; Luoma, H; Jousimies-Somer, H; Pyhälä, L; Asikainen, S; Alaluusua, S

    1995-01-01

    The susceptibilities of 379 clinical mutans streptococcal isolates to chlorhexidine (CHX) were tested by agar dilution according to the standards of the National Committee for Clinical Laboratory Standards. Isolates were obtained from saliva samples of 34 young mothers who had high or moderate salivary levels of mutans streptococci at baseline. Samples were collected on three occasions, before childbirth, when each child was 6 months old, and 1 year later. Of these isolates, 50% were inhibited at 1 microgram of CHX per ml, 90% were inhibited at 2.0 micrograms/ml, and all were inhibited at 4.0 micrograms/ml. The MICs for Streptococcus mutans isolates (serotypes c, e, and f) were lower than those for Streptococcus sobrinus isolates (serotypes d and g). In some subjects, the MICs for isolates of the same serotype were different. This phenomenon was studied by ribotyping isolates (n = 45) from selected subjects (n = 7). It was found that if there were intraindividual differences in the MICs for isolates of the same serotype, then the ribotypes of these isolates were different. In order to decrease the mutans streptococcal infection risk for children, 24 mothers (test group) brushed their teeth periodically with a gel that contained 0.3% CHX digluconate and 0.2% NaF, pH 5.8, between the second and third sampling occasions. The gel was used twice a day for the first 10 days of each month. Development of resistant strains during CHX-NaF gel use was not detected. The serotype distribution of isolates from the test group after 1 year of periodic CHX-NaF gel use did not differ from that at baseline. Periodic CHX-NaF gel brushing did not lead to lower salivary mutans streptococcal counts. PMID:7785991

  5. Mannanoligosaccharide agglutination by Salmonella enterica strains isolated from carrier pigs

    PubMed Central

    Borowsky, Luciane; Corção, Gertrudes; Cardoso, Marisa

    2009-01-01

    Type-1 fimbriae are associated with most Salmonella enterica serovars and are an essential factor for host colonization. Mannanoligosaccharides (MOS), a prebiotic that is agglutinated by type-1 fimbriae, are proposed for the control of enterobacteria colonization and may be an alternative to Salmonella control in pigs. The aim of this study was to evaluate the capability of porcine Salmonella strains to adhere to MOS in vitro. A total of 108 strains of Salmonella sp. isolated from carrier pigs were evaluated for the amplification of fimA and fimH genes, agglutination of MOS and hemagglutination. In all tested strains, amplicons of expected size were detected for both fimA and fimH gene. In the hemagglutination assays, 31 (28.7%) strains presented mannose–sensitive agglutination of erythrocytes, indicating that the strains were expressing type-1 fimbriae. Considering only strains expressing the type-1 fimbriae, 23 (74.2%) presented a strong agglutination of MOS, 3 (9.6%) a weak reaction and 5 (16.2%) none. The results indicate that Salmonella enterica strains expressing type-1 fimbriae can agglutinate effectively in vitro to MOS. PMID:24031388

  6. Capsular polysaccharide antigens for detection of serotype-specific antibodies to Actinobacillus pleuropneumoniae.

    PubMed Central

    Bossé, J T; Johnson, R P; Rosendal, S

    1990-01-01

    Capsular polysaccharides (CPS) of serotypes 1, 2, 5 and 7 of Actinobacillus (Haemophilus) pleuropneumoniae were obtained from 18 h culture supernatants by precipitation with hexadecyltrimethyl-ammonium bromide (Cetavlon) followed by extraction with sodium chloride and reprecipitation in ethanol. These crude extracts, and portions purified further by phenol extraction to remove contaminating proteins, were evaluated as antigens for the detection of serotype-specific antibodies to A. pleuropneumoniae in sera from immunized rabbits and swine by an enzyme-linked immunosorbent assay. The crude extracts reacted strongly with homologous antisera, but except for serotype 1 showed considerable cross-reactivity with antisera to other serotypes. Phenol extraction greatly improved the serospecificity of the antigens from serotypes 1, 7 and, to a lesser extent, 5. The serotype 2 CPS antigen showed poor reactivity following phenol extraction, and did not appear as useful for detection of serotype-specific antibodies. PMID:2379111

  7. Changing Pattern of Dengue Virus Serotypes in Thailand between 2004 and 2010

    PubMed Central

    Pongsiri, Piyathida; Themboonlers, Apiradee

    2012-01-01

    Dengue virus infection is a major concern in several countries, and more than 50 million people are infected worldwide each year. Thailand is one of the countries where people are susceptible to infection due to favourable geographical and environmental conditions. In this retrospective study, we reported the changing pattern of dengue virus serotypes during the period between 2004 and 2010. The following percentage prevalence showed different serotypes of dengue virus (DENV) predominant in respective years: DENV1 in 2004 (56.41%), DENV4 in 2007 (50%), DENV1 in 2008 (57.41%), and DENV3 in 2010 (38.7%). Moreover, the major serotypes were not stable as they showed a shift from one serotype to another. We also found co-infection with two different serotypes and reported the clinical manifestations, which were not different from infection with a single serotype. Co-infection with various serotypes may not necessarily cause more severe disease. PMID:23082638

  8. Immunoblot Method To Detect Streptococcus pneumoniae and Identify Multiple Serotypes from Nasopharyngeal Secretions

    PubMed Central

    Bronsdon, Melinda A.; O'Brien, Katherine L.; Facklam, Richard R.; Whitney, Cynthia G.; Schwartz, Benjamin; Carlone, George M.

    2004-01-01

    Conventional culture techniques are limited in the ability to detect multiple Streptococcus pneumoniae serotypes in nasopharyngeal (NP) secretions. We developed an immunoblot (IB) method with monoclonal antibodies (MAbs) to detect S. pneumoniae and to identify serotypes. NP specimens stored in skim milk-tryptone-glucose-glycerol medium were assessed by the IB method and the reference culture method (RM). In the RM, four optochin-sensitive alpha-hemolytic colonies resembling pneumococci were typed by the Quellung reaction. In the IB method, a nitrocellulose membrane blot of surface growth was reacted with a pneumococcal surface adhesion (PsaA) MAb and visualized. Of 47 NP specimens, 32 (68%) were found to be positive and 13 (28%) were found to be negative for pneumococci by both methods; each method alone yielded one positive result. The sensitivity and specificity of the IB method for the detection of pneumococci were 97 and 93%, respectively. To identify serotypes, blots were tested with serotype-specific MAbs (4, 6A, 6B, 9V, 14, 18C, 19F, and 23F). To detect the remaining serotypes, positive serotype-specific replicate blots were compared visually to an original anti-PsaA-positive blot; four unidentified colonies were subcultured and serotyped by the Quellung reaction. Fifty-eight S. pneumoniae-positive NP specimens containing 69 pneumococcal strains (23 serotypes) were tested; 68 (98.6%) of the strains were detected by the IB method, and 66 (95.6%) were detected by the RM. For 11 specimens found to contain two serotypes, both methods detected both serotypes in 7 (63.6%), the IB method alone detected the two serotypes in 3 (27.3%), and the RM alone detected both serotypes in 1 (9%). The IB method identified multiple clones and minor populations of pneumococci in NP secretions. This method is useful for detecting specific serotypes and carriage of multiple serotypes in epidemiologic surveillance and carriage studies. PMID:15071010

  9. Molecular identification of Salmonella enterica subsp. enterica serovar Gallinarum biovars Gallinarum and Pullorum by a duplex PCR assay.

    PubMed

    Batista, Diego Felipe Alves; de Freitas Neto, Oliveiro Caetano; de Almeida, Adriana Maria; Barrow, Paul Andrew; de Oliveira Barbosa, Fernanda; Berchieri Junior, Angelo

    2016-07-01

    Salmonella enterica subsp. enterica serovar Gallinarum biovar Gallinarum (S Gallinarum) and biovar Pullorum (S Pullorum) are 2 poultry pathogens that cause major economic losses to the poultry industry worldwide. Control of both diseases mainly relies on the adoption of biosecurity programs, and success is dependent on accurate and fast detection. Based on this concept, we developed a duplex PCR assay, targeting 2 chromosomal sequences, which allowed us to precisely identify and differentiate S Gallinarum and S Pullorum field strains. This assay was validated by testing genomic DNA from 40 S Gallinarum and 29 S Pullorum field strains, 87 other Salmonella serovars, and 7 non-Salmonella strains. The serovar identifier region (SIR) primers produced a fragment only in S Gallinarum and S Pullorum strains, whereas the fragment from the ratA coding sequence, which was previously demonstrated to differentiate the 2 biovars, was also amplified from other Salmonella serovars. Our results showed that the combination of both SIR and ratA amplifications could be used to identify as well as to differentiate colonies of S Gallinarum and S Pullorum reliably. Thus, we believe this methodology can be a useful ancillary tool for routine veterinary diagnostic laboratories by providing rapid, accurate results. PMID:27216724

  10. Intestinal Cytokine Responses to Salmonella enterica Serovar Typhimurium Infection in Young Chicks

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Salmonella enterica serovar typhimurium is one of the most frequently isolated strains in human salmonellosis worldwide, and is commonly found in broilers. Successful prevention and control of Salmonella colonization in poultry require better understanding of intestinal mucosal immune response to ...

  11. Characterization of the novel T4-like Salmonella enterica bacteriophage STP4-a and its endolysin.

    PubMed

    Li, Meng; Li, Mengzhe; Lin, Hong; Wang, Jingxue; Jin, Yanqiu; Han, Feng

    2016-02-01

    While screening for new antimicrobial agents for multidrug-resistant Salmonella enterica, the novel lytic bacteriophage STP4-a was isolated and characterized. Phage morphology revealed that STP4-a belongs to the family Myoviridae. Bacterial challenge assays showed that different serovars of Salmonella enterica were susceptible to STP4-a infection. The genomic characteristics of STP4-a, containing 159,914 bp of dsDNA with an average GC content of 36.86 %, were determined. Furthermore, the endolysin of STP4-a was expressed and characterized. The novel endolysin, LysSTP4, has hydrolytic activity towards outer-membrane-permeabilized S. enterica and Escherichia coli. These results provide essential information for the development of novel phage-based biocontrol agents against S. enterica. PMID:26563319

  12. Differential Responses of Macrophages to Salmonella enterica Serovars Enteritidis and Typhimurium

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Macrophages are major effectors against Salmonella infection, and also transport bacteria between host tissues and provide a protected site for intracellular bacterial replication. We hypothesized that differences in chicken macrophage responses to Salmonella enterica serovar Enteritidis (SE) and s...

  13. Analysis of Plasmid and Chromosomal DNA of Multidrug-Resistant Salmonella enterica Serovar Typhi from Asia

    PubMed Central

    Mirza, S.; Kariuki, S.; Mamun, K. Z.; Beeching, N. J.; Hart, C. A.

    2000-01-01

    Molecular analysis of chromosomal DNA from 193 multidrug-resistant (MDR) Salmonella enterica serovar Typhi isolates from 1990 to 1995 from Pakistan, Kuwait, Malaysia, Bangladesh, and India produced a total of five major different pulsed-field gel electrophoresis (PFGE) patterns. Even within a particular country MDR S. enterica serovar Typhi DNA was found to be in different PFGE groups. Similar self-transferable 98-MDa plasmids belonging to either incompatibility group incHI1 or incHI1/FIIA were implicated in the MDR phenotype in S. enterica serovar Typhi isolates from all the locations except Quetta, Pakistan, where the majority were of incFIA. A total of five different PFGE genotypes with six different plasmids, based on incompatibility and restriction endonuclease analysis groups, were found among these MDR S. enterica serovar Typhi isolates. PMID:10747124

  14. Evaluation of corn oil as an additive in the pre-enrichment step to increase recovery of Salmonella enterica from oregano.

    PubMed

    Jean-Gilles Beaubrun, Junia; Flamer, Marie-Laure; Addy, Nicole; Ewing, Laura; Gopinath, Gopal; Jarvis, Karen; Grim, Chris; Hanes, Darcy E

    2016-08-01

    Phenolic compounds associated with essential oils of spices and herbs possess a variety of antioxidant and antimicrobial properties that interfere with Salmonella detection from fresh and dried products. Finding a compound to neutralize the effect of these antimicrobial compounds, while allowing Salmonella growth during pre-enrichment, is a crucial step in both traditional pathogen isolation and molecular detection from these foods. This study evaluated the effectiveness of corn oil as a component of the pre-enrichment broth to counteract antimicrobial compounds properties and increase the recovery of Salmonella from spices. Oregano samples artificially contaminated with Salmonella enterica were pre-enriched in modified Buffered Peptone Water (mBPW) supplemented with and without 2% (vol/vol) corn oil respectively. Samples were incubated overnight at 37 °C. The results showed that recovery of Salmonella from oregano samples was increased by ≥50% when pre-enriched with corn oil. Serovars were confirmed using a PCR serotyping method. In addition, shot-gun metagenomics analyses demonstrated bacterial diversity and the effect of corn oil on the relative prevalence of Salmonella in the oregano samples. Modifying pre-enrichment broths with corn oil improved the detection and isolation of Salmonella from oregano, and may provide an alternative method for pathogen detection in dried food matrices such as spices. PMID:27052719

  15. National Outbreak of Salmonella Serotype Saintpaul Infections: Importance of Texas Restaurant Investigations in Implicating Jalapeño Peppers

    PubMed Central

    Mody, Rajal K.; Greene, Sharon A.; Gaul, Linda; Sever, Adrianne; Pichette, Sarah; Zambrana, Ingrid; Dang, Thi; Gass, Angie; Wood, René; Herman, Karen; Cantwell, Laura B.; Falkenhorst, Gerhard; Wannemuehler, Kathleen; Hoekstra, Robert M.; McCullum, Isaac; Cone, Amy; Franklin, Lou; Austin, Jana; Delea, Kristin; Behravesh, Casey Barton; Sodha, Samir V.; Yee, J. Christopher; Emanuel, Brian; Al-Khaldi, Sufian F.; Jefferson, Val; Williams, Ian T.; Griffin, Patricia M.; Swerdlow, David L.

    2011-01-01

    Background In May 2008, PulseNet detected a multistate outbreak of Salmonella enterica serotype Saintpaul infections. Initial investigations identified an epidemiologic association between illness and consumption of raw tomatoes, yet cases continued. In mid-June, we investigated two clusters of outbreak strain infections in Texas among patrons of Restaurant A and two establishments of Restaurant Chain B to determine the outbreak's source. Methodology/Principal Findings We conducted independent case-control studies of Restaurant A and B patrons. Patients were matched to well controls by meal date. We conducted restaurant environmental investigations and traced the origin of implicated products. Forty-seven case-patients and 40 controls were enrolled in the Restaurant A study. Thirty case-patients and 31 controls were enrolled in the Restaurant Chain B study. In both studies, illness was independently associated with only one menu item, fresh salsa (Restaurant A: matched odds ratio [mOR], 37; 95% confidence interval [CI], 7.2–386; Restaurant B: mOR, 13; 95% CI 1.3–infinity). The only ingredient in common between the two salsas was raw jalapeño peppers. Cultures of jalapeño peppers collected from an importer that supplied Restaurant Chain B and serrano peppers and irrigation water from a Mexican farm that supplied that importer with jalapeño and serrano peppers grew the outbreak strain. Conclusions/Significance Jalapeño peppers, contaminated before arrival at the restaurants and served in uncooked fresh salsas, were the source of these infections. Our investigations, critical in understanding the broader multistate outbreak, exemplify an effective approach to investigating large foodborne outbreaks. Additional measures are needed to reduce produce contamination. PMID:21373185

  16. Formal Comment to Pettengill: The Time to Most Recent Common Ancestor Does Not (Usually) Approximate the Date of Divergence

    PubMed Central

    Achtman, Mark; Zhou, Zhemin; Didelot, Xavier

    2015-01-01

    In 2013 Zhou et al. concluded that Salmonella enterica serovar Agona represents a genetically monomorphic lineage of recent ancestry, whose most recent common ancestor existed in 1932, or earlier. The Abstract stated ‘Agona consists of three lineages with minimal mutational diversity: only 846 single nucleotide polymorphisms (SNPs) have accumulated in the non-repetitive, core genome since Agona evolved in 1932 and subsequently underwent a major population expansion in the 1960s.’ These conclusions have now been criticized by Pettengill, who claims that the evolutionary models used to date Agona may not have been appropriate, the dating estimates were inaccurate, and the age of emergence of Agona should have been qualified by an upper limit reflecting the date of its divergence from an outgroup, serovar Soerenga. We dispute these claims. Firstly, Pettengill’s analysis of Agona is not justifiable on technical grounds. Secondly, an upper limit for divergence from an outgroup would only be meaningful if the outgroup were closely related to Agona, but close relatives of Agona are yet to be identified. Thirdly, it is not possible to reliably date the time of divergence between Agona and Soerenga. We conclude that Pettengill’s criticism is comparable to a tempest in a teapot. PMID:26274924

  17. Formal Comment to Pettengill: The Time to Most Recent Common Ancestor Does Not (Usually) Approximate the Date of Divergence.

    PubMed

    Achtman, Mark; Zhou, Zhemin; Didelot, Xavier

    2015-01-01

    In 2013 Zhou et al. concluded that Salmonella enterica serovar Agona represents a genetically monomorphic lineage of recent ancestry, whose most recent common ancestor existed in 1932, or earlier. The Abstract stated 'Agona consists of three lineages with minimal mutational diversity: only 846 single nucleotide polymorphisms (SNPs) have accumulated in the non-repetitive, core genome since Agona evolved in 1932 and subsequently underwent a major population expansion in the 1960s.' These conclusions have now been criticized by Pettengill, who claims that the evolutionary models used to date Agona may not have been appropriate, the dating estimates were inaccurate, and the age of emergence of Agona should have been qualified by an upper limit reflecting the date of its divergence from an outgroup, serovar Soerenga. We dispute these claims. Firstly, Pettengill's analysis of Agona is not justifiable on technical grounds. Secondly, an upper limit for divergence from an outgroup would only be meaningful if the outgroup were closely related to Agona, but close relatives of Agona are yet to be identified. Thirdly, it is not possible to reliably date the time of divergence between Agona and Soerenga. We conclude that Pettengill's criticism is comparable to a tempest in a teapot. PMID:26274924

  18. Pork Meat as a Potential Source of Salmonella enterica subsp. arizonae Infection in Humans

    PubMed Central

    Kritas, Spyridon; Govaris, Alexander; Burriel, Angeliki R.

    2014-01-01

    Salmonella enterica subsp. arizonae was isolated from 13 of 123 slaughtered pigs in central Greece. The samples cultured were feces, ileum tissue, mesenteric lymph nodes, and gallbladder swabs. A total of 74 isolates from 492 samples were identified as Salmonella spp. by use of standard laboratory culture media and two commercial micromethods and by use of a polyvalent slide agglutination test for the detection of O and H antigens. Among them were 19 (25.68%) suspected to be S. enterica subsp. arizonae according to analysis with standard laboratory culture media. Of those, 14 were identified as S. enterica subsp. arizonae by the API 20E (bioMérieux, France) and the Microgen GnA+B-ID (Microgen Bioproducts, Ltd., United Kingdom) identification systems. All the isolates were tested for resistance to 23 antimicrobials. Strains identified as S. enterica subsp. arizonae were resistant to 17 (70.8%) antibiotics. The highest proportions of resistance were observed for sulfamethoxazole-trimethoprim (71.4%), tetracycline (71.4%), ampicillin (64.3%), and amoxicillin (57.1%). Two isolates were resistant to aztreonam (7.1%) and tigecycline (7.1%), used only for the treatment of humans. Thus, pork meat may play a role in the transmission of antibiotic-resistant S. enterica subsp. arizonae to human consumers. This is the first report of S. enterica subsp. arizonae isolation from pigs. PMID:24335956

  19. PCR-based identification of serotype 2 isolates of Actinobacillus pleuropneumoniae biovars I and II.

    PubMed

    Hüssy, Daniela; Schlatter, Yvonne; Miserez, Raymond; Inzana, Thomas; Frey, Joachim

    2004-04-19

    A genetic typing method utilizing PCR for the identification of Actinobacillus pleuropneumoniae serotype 2 isolates has been developed based on the in vitro amplification of a 1.4 kb DNA segment of the serotype 2 capsular polysaccharide genes cps2AB. The assay was tested with all serotype reference strains and a collection of 92 different A. pleuropneumoniae strains of all 15 serotypes of both biovars I and II, originating from 18 different countries worldwide. The cps2 based PCR identified the serotype 2 reference strain and all 12 serotype 2 collection strains contained in this set. DNA was not amplified from the remaining A. pleuropneumoniae reference and collection strains, indicating the PCR assay was highly specific. Furthermore, the PCR method detected all 31 A. pleuropneumoniae serotype 2 field isolates from diseased pigs that were identified in parallel as serotype 2 by agar gel diffusion. The serotype 2 PCR assay proved to be highly specific and reliable for the identification of serotype 2 isolates of A. pleuropneumoniae. PMID:15066734

  20. Draft Genome Sequences of Listeria monocytogenes Serotype 4b Strains 944 and 2993 and Serotype 1/2c Strains 198 and 2932.

    PubMed

    Casey, Aidan; McAuliffe, Olivia; Fox, Edward M; Leong, Dara; Gahan, Cormac G M; Jordan, Kieran

    2016-01-01

    Listeria monocytogenes is a foodborne pathogen and the causative agent of listeriosis among humans and animals. The draft genome sequences of L. monocytogenes serotype 4b strains 944 and 2993 and serotype 1/2c strains 198 and 2932 are reported here. PMID:27257200

  1. Draft Genome Sequences of Listeria monocytogenes Serotype 4b Strains 944 and 2993 and Serotype 1/2c Strains 198 and 2932

    PubMed Central

    Casey, Aidan; Fox, Edward M.; Leong, Dara; Gahan, Cormac G. M.; Jordan, Kieran

    2016-01-01

    Listeria monocytogenes is a foodborne pathogen and the causative agent of listeriosis among humans and animals. The draft genome sequences of L. monocytogenes serotype 4b strains 944 and 2993 and serotype 1/2c strains 198 and 2932 are reported here. PMID:27257200

  2. Serotype-specific differences in short- and longer-term mortality following invasive pneumococcal disease.

    PubMed

    Hughes, G J; Wright, L B; Chapman, K E; Wilson, D; Gorton, R

    2016-09-01

    Invasive pneumococcal disease (IPD), caused by infection with Streptococcus pneumoniae, has a substantial global burden. There are over 90 known serotypes of S. pneumoniae with a considerable body of evidence supporting serotype-specific mortality rates immediately following IPD. This is the first study to consider the association between serotype and longer-term mortality following IPD. Using enhanced surveillance data from the North East of England we assessed both the short-term (30-day) and longer-term (⩽7 years) independent adjusted associations between individual serotypes and mortality following IPD diagnosis using logistic regression and extended Cox proportional hazards models. Of the 1316 cases included in the analysis, 243 [18·5%, 95% confidence interval (CI) 16·4-20·7] died within 30 days of diagnosis. Four serotypes (3, 6A, 9N, 19 F) were significantly associated with overall increased 30-day mortality. Effects were observable only for older adults (⩾60 years). After extension of the window to 12 months and 36 months, one serotype was associated with significantly increased mortality at 12 months (19 F), but no individual serotypes were associated with increased mortality at 36 months. Two serotypes had statistically significant hazard ratios (HR) for longer-term mortality: serotype 1 for reduced mortality (HR 0·51, 95% CI 0·30-0·86) and serotype 9N for increased mortality (HR 2·30, 95% CI 1·29-4·37). The association with serotype 9N was no longer observed after limiting survival analysis to an observation period starting 30 days after diagnosis. This study supports the evidence for associations between serotype and short-term (30-day) mortality following IPD and provides the first evidence for the existence of statistically significant associations between individual serotypes and longer-term variation in mortality following IPD. PMID:27193457

  3. Salmonella enterica flagellin is recognized via FLS2 and activates PAMP-triggered immunity in Arabidopsis thaliana.

    PubMed

    Garcia, Ana Victoria; Charrier, Amélie; Schikora, Adam; Bigeard, Jean; Pateyron, Stephanie; de Tauzia-Moreau, Marie-Ludivine; Evrard, Alexandre; Mithöfer, Axel; Martin-Magniette, Marie Laure; Virlogeux-Payant, Isabelle; Hirt, Heribert

    2014-04-01

    Infections with Salmonella enterica belong to the most prominent causes of food poisoning and infected fruits and vegetables represent important vectors for salmonellosis. Recent evidence indicates that plants recognize S. enterica and raise defense responses. Nonetheless, the molecular mechanisms controlling the interaction of S. enterica with plants are still largely unclear. Here, we show that flagellin from S. enterica represents a prominent pathogen-associated molecular pattern (PAMP) in Arabidopsis thaliana, which induces PAMP-triggered immunity (PTI) via the recognition of the flg22 domain by the receptor kinase FLS2. The Arabidopsis fls2 mutant shows reduced though not abolished PTI activation, indicating that plants rely also on recognition of other S. enterica PAMPs. Interestingly, the S. enterica type III secretion system (T3SS) mutant prgH- induced stronger defense gene expression than wild-type bacteria in Arabidopsis, suggesting that T3SS effectors are involved in defense suppression. Furthermore, we observe that S. enterica strains show variation in the flg22 epitope, which results in proteins with reduced PTI-inducing activity. Altogether, these results show that S. enterica activates PTI in Arabidopsis and suggest that, in order to accomplish plant colonization, S. enterica evolved strategies to avoid or suppress PTI. PMID:24198231

  4. Common occurrence of concurrent infections by multiple dengue virus serotypes.

    PubMed

    Loroño-Pino, M A; Cropp, C B; Farfán, J A; Vorndam, A V; Rodríguez-Angulo, E M; Rosado-Paredes, E P; Flores-Flores, L F; Beaty, B J; Gubler, D J

    1999-11-01

    The co-circulation of all 4 dengue virus serotypes in the same community, common since the 1950s in Southeast Asia, has now become a frequent occurrence in many Caribbean Islands, Mexico, and Central and South America in the past 20 years. As a consequence, the frequency of concurrent infections would be expected to increase in these areas. To assess this, using state of the art technology, we screened viremic serum samples and mosquitoes inoculated with serum samples collected during epidemics involving multiple dengue virus serotypes in Indonesia, Mexico, and Puerto Rico for virus isolation. Of 292 samples tested, 16 (5.5%) were found to contain 2 or more dengue viruses by an indirect immunofluorescence test and/or the reverse transcriptase-polymerase chain reaction. PMID:10586902

  5. Adenovirus serotype 5 hexon mediates liver gene transfer.

    PubMed

    Waddington, Simon N; McVey, John H; Bhella, David; Parker, Alan L; Barker, Kristeen; Atoda, Hideko; Pink, Rebecca; Buckley, Suzanne M K; Greig, Jenny A; Denby, Laura; Custers, Jerome; Morita, Takashi; Francischetti, Ivo M B; Monteiro, Robson Q; Barouch, Dan H; van Rooijen, Nico; Napoli, Claudio; Havenga, Menzo J E; Nicklin, Stuart A; Baker, Andrew H

    2008-02-01

    Adenoviruses are used extensively as gene transfer agents, both experimentally and clinically. However, targeting of liver cells by adenoviruses compromises their potential efficacy. In cell culture, the adenovirus serotype 5 fiber protein engages the coxsackievirus and adenovirus receptor (CAR) to bind cells. Paradoxically, following intravascular delivery, CAR is not used for liver transduction, implicating alternate pathways. Recently, we demonstrated that coagulation factor (F)X directly binds adenovirus leading to liver infection. Here, we show that FX binds to the Ad5 hexon, not fiber, via an interaction between the FX Gla domain and hypervariable regions of the hexon surface. Binding occurs in multiple human adenovirus serotypes. Liver infection by the FX-Ad5 complex is mediated through a heparin-binding exosite in the FX serine protease domain. This study reveals an unanticipated function for hexon in mediating liver gene transfer in vivo. PMID:18267072

  6. Neisseria meningitidis: serotyping and subtyping by whole cell ELISA.

    PubMed

    Prakash, K; Lakshmy, A; Malhotra, V L

    1993-09-01

    Twenty strains of Neisseria meningitidis isolated from clinically diagnosed cases of meningococcal disease were subjected to serogrouping, employing slide agglutination followed by serotyping and serosubtyping by whole cell ELISA using monoclonal typing antisera. All isolates were from sporadic cases of meningitis during a period of two years from various hospitals in Delhi. All 20 isolates were grouped as serogroup A and typed as serotype 4, except one strain which was untypable. On serosubtyping the isolates were found to belong to P1.9 (7 strains) followed by P1.1 (5), P1.9 (2), P1.16,1 (2), P1.6,10 (2), P1.10,7,1 (1) and non-subtypable (1). PMID:8241833

  7. The structural elucidation of the Salmonella enterica subsp. enterica, reveals that it contains both O-factors 4 and 5 on the LPS antigen.

    PubMed

    De Castro, Cristina; Lanzetta, Rosa; Leone, Serena; Parrilli, Michelangelo; Molinaro, Antonio

    2013-04-01

    Spectroscopic investigation of the O-antigen from Salmonella enterica subsp. enterica revealed fine details on the acetylation pattern, the biological repeating unit and the polymerization degree. Acetylation at O-2 of the abequose residue, defined both O-factors 4 and 5 in the O-antigen chain of the lipopolysaccharide. NMR observation of the terminal non-reducing end of the polymer confirmed previous data regarding the biological repeating unit and showed an average polymerization degree of 5. The information about these structural elements might contribute to the understanding of key features of the biology of this pathogen, as phase variation and/or adaptation to the external environment. PMID:23419941

  8. Virulence Gene Regulation by l-Arabinose in Salmonella enterica

    PubMed Central

    López-Garrido, Javier; Puerta-Fernández, Elena; Cota, Ignacio; Casadesús, Josep

    2015-01-01

    Invasion of the intestinal epithelium is a critical step in Salmonella enterica infection and requires functions encoded in the gene cluster known as Salmonella Pathogenicity Island 1 (SPI-1). Expression of SPI-1 genes is repressed by l-arabinose, and not by other pentoses. Transport of l-arabinose is necessary to repress SPI-1; however, repression is independent of l-arabinose metabolism and of the l-arabinose-responsive regulator AraC. SPI-1 repression by l-arabinose is exerted at a single target, HilD, and the mechanism appears to be post-translational. As a consequence of SPI-1 repression, l-arabinose reduces translocation of SPI-1 effectors to epithelial cells and decreases Salmonella invasion in vitro. These observations reveal a hitherto unknown role of l-arabinose in gene expression control and raise the possibility that Salmonella may use L-arabinose as an environmental signal. PMID:25991823

  9. Gene Transfer between Salmonella enterica Serovar Typhimurium inside Epithelial Cells

    PubMed Central

    Ferguson, Gayle C.; Heinemann, Jack A.; Kennedy, Martin A.

    2002-01-01

    Virulence and antibiotic resistance genes transfer between bacteria by bacterial conjugation. Conjugation also mediates gene transfer from bacteria to eukaryotic organisms, including yeast and human cells. Predicting when and where genes transfer by conjugation could enhance our understanding of the risks involved in the release of genetically modified organisms, including those being developed for use as vaccines. We report here that Salmonella enterica serovar Typhimurium conjugated inside cultured human cells. The DNA transfer from donor to recipient bacteria was proportional to the probability that the two types of bacteria occupied the same cell, which was dependent on viable and invasive bacteria and on plasmid tra genes. Based on the high frequencies of gene transfer between bacteria inside human cells, we suggest that such gene transfers occur in situ. The implications of gene transfer between bacteria inside human cells, particularly in the context of antibiotic resistance, are discussed. PMID:11914355

  10. Osteomyelitis caused by Salmonella enterica serovar derby in boa constrictor.

    PubMed

    de Souza, Suyene O; Casagrande, Renata A; Guerra, Priscila R; Cruz, Cláudio E F; Veit, Evandro; Cardoso, Marisa R I; Driemeier, David

    2014-09-01

    After demonstrating chronic weight loss, prostration, and muscle flaccidness, a captive-bred 9-mo-old boa constrictor (Boa constrictor constrictor) died and was submitted for necropsy. Along the spinal column there were multiple, yellowish white, macroscopic nodules of 1-5 mm in diameter in the ventral side of the vertebral body and in the intervertebral spaces. Severe multifocal necrotizing osteomyelitis associated with granulomatous inflammation was the main histologic finding in the vertebral column. In the liver, there was discrete but similar granulomatous changes. Positive anti-Salmonella immunostaining was observed in the spinal column and in the liver. Salmonella enterica serovar Derby was isolated from fragments of the spinal column. These bacteria are important cause of disease in captive reptiles. PMID:25314834

  11. Cytotoxic T cell adjuvant effects of three Salmonella enterica flagellins

    PubMed Central

    Braga, Catarina J.M.; Massis, Liliana M.; Alencar, Bruna C.G.; Rodrigues, Maurício M.; Sbrogio-Almeida, M.E.; Ferreira, Luís C.S.

    2008-01-01

    Bacterial flagellins are important virulence-associated factors and strong inducers of inflammatory responses in mammalian hosts. Flagellins have also been investigated as potential vaccine adjuvants, either for induction of humoral or cellular immune responses, to different target antigens. In this study we investigated the adjuvant properties of three Salmonella enterica flagellins types (FliCd, FliCi and FljB) to an ovalbumin-derived CD8+ T cell-restricted epitope (OVA257–264). Although mice immunized with the three tested flagellins elicited antigen-specific activated CD8+ T cells, only animals immunized with FliCi and FliCd flagellins admixed with ovalbumin mounted specific in vivo cytotoxic responses to peptide-pulsed target cells. The present results indicate that Salmonella flagellins are endowed with type-specific adjuvant effects toward murine CD8+ T cells, a feature that may impact their use as adjuvants for prophylatic or therapeutic vaccines. PMID:24031176

  12. Flagella-independent surface motility in Salmonella enterica serovar Typhimurium

    PubMed Central

    Park, Sun-Yang; Pontes, Mauricio H.; Groisman, Eduardo A.

    2015-01-01

    Flagella are multiprotein complexes necessary for swimming and swarming motility. In Salmonella enterica serovar Typhimurium, flagella-mediated motility is repressed by the PhoP/PhoQ regulatory system. We now report that Salmonella can move on 0.3% agarose media in a flagella-independent manner when experiencing the PhoP/PhoQ-inducing signal low Mg2+. This motility requires the PhoP-activated mgtA, mgtC, and pagM genes, which specify a Mg2+ transporter, an inhibitor of Salmonella’s own F1Fo ATPase, and a small protein of unknown function, respectively. The MgtA and MgtC proteins are necessary for pagM expression because pagM mRNA levels were lower in mgtA and mgtC mutants than in wild-type Salmonella, and also because pagM expression from a heterologous promoter rescued motility in mgtA and mgtC mutants. PagM promotes group motility by a surface protein(s), as a pagM-expressing strain conferred motility upon a pagM null mutant, and proteinase K treatment eliminated motility. The pagM gene is rarely found outside subspecies I of S. enterica and often present in nonfunctional allelic forms in organisms lacking the identified motility. Deletion of the pagM gene reduced bacterial replication on 0.3% agarose low Mg2+ media but not in low Mg2+ liquid media. Our findings define a form of motility that allows Salmonella to scavenge nutrients and to escape toxic compounds in low Mg2+ semisolid environments. PMID:25624475

  13. Quinolone Resistance Mechanisms Among Salmonella enterica in Malaysia.

    PubMed

    Thong, Kwai Lin; Ngoi, Soo Tein; Chai, Lay Ching; Teh, Cindy Shuan Ju

    2016-06-01

    The prevalence of quinolone-resistant Salmonella enterica is on the rise worldwide. Salmonella enterica is one of the major foodborne pathogens in Malaysia. Therefore, we aim to investigate the occurrence and mechanisms of quinolone resistance among Salmonella strains isolated in Malaysia. A total of 283 Salmonella strains isolated from food, humans, and animals were studied. The disk diffusion method was used to examine the quinolone susceptibility of the strains, and the minimum inhibitory concentration (MIC) values of nalidixic acid and ciprofloxacin were also determined. DNA sequencing of the quinolone resistance-determining regions (QRDRs) of gyrase and topoisomerase IV genes and the plasmid-borne qnr genes was performed. The transfer of the qnr gene was examined through transconjugation experiment. A total of 101 nalidixic acid-resistant Salmonella strains were identified. In general, all strains were highly resistant to nalidixic acid (average MICNAL, 170 μg/ml). Resistance to ciprofloxacin was observed in 30.7% of the strains (1 ≤ MICCIP ≤ 2 μg/ml). Majority of the strains contained missense mutations in the QRDR of gyrA (69.3%). Silent mutations were frequently detected in gyrB (75.2%), parC (27.7%), and parE (51.5%) within and beyond the QRDRs. Novel mutations were detected in parC and parE. The plasmid-borne qnrS1 variant was found in 36.6% of the strains, and two strains were found to be able to transfer the qnrS1 gene. Overall, mutations in gyrA and the presence of qnrS1 genes might have contributed to the high level of quinolone resistance among the strains. Our study provided a better understanding on the status of quinolone resistance among Salmonella strains circulating in Malaysia. PMID:26683630

  14. Flagella-independent surface motility in Salmonella enterica serovar Typhimurium.

    PubMed

    Park, Sun-Yang; Pontes, Mauricio H; Groisman, Eduardo A

    2015-02-10

    Flagella are multiprotein complexes necessary for swimming and swarming motility. In Salmonella enterica serovar Typhimurium, flagella-mediated motility is repressed by the PhoP/PhoQ regulatory system. We now report that Salmonella can move on 0.3% agarose media in a flagella-independent manner when experiencing the PhoP/PhoQ-inducing signal low Mg(2+). This motility requires the PhoP-activated mgtA, mgtC, and pagM genes, which specify a Mg(2+) transporter, an inhibitor of Salmonella's own F1Fo ATPase, and a small protein of unknown function, respectively. The MgtA and MgtC proteins are necessary for pagM expression because pagM mRNA levels were lower in mgtA and mgtC mutants than in wild-type Salmonella, and also because pagM expression from a heterologous promoter rescued motility in mgtA and mgtC mutants. PagM promotes group motility by a surface protein(s), as a pagM-expressing strain conferred motility upon a pagM null mutant, and proteinase K treatment eliminated motility. The pagM gene is rarely found outside subspecies I of S. enterica and often present in nonfunctional allelic forms in organisms lacking the identified motility. Deletion of the pagM gene reduced bacterial replication on 0.3% agarose low Mg(2+) media but not in low Mg(2+) liquid media. Our findings define a form of motility that allows Salmonella to scavenge nutrients and to escape toxic compounds in low Mg(2+) semisolid environments. PMID:25624475

  15. Advances in Molecular Serotyping and Subtyping of Escherichia coli†

    PubMed Central

    Fratamico, Pina M.; DebRoy, Chitrita; Liu, Yanhong; Needleman, David S.; Baranzoni, Gian Marco; Feng, Peter

    2016-01-01

    Escherichia coli plays an important role as a member of the gut microbiota; however, pathogenic strains also exist, including various diarrheagenic E. coli pathotypes and extraintestinal pathogenic E. coli that cause illness outside of the GI-tract. E. coli have traditionally been serotyped using antisera against the ca. 186 O-antigens and 53 H-flagellar antigens. Phenotypic methods, including bacteriophage typing and O- and H- serotyping for differentiating and characterizing E. coli have been used for many years; however, these methods are generally time consuming and not always accurate. Advances in next generation sequencing technologies have made it possible to develop genetic-based subtyping and molecular serotyping methods for E. coli, which are more discriminatory compared to phenotypic typing methods. Furthermore, whole genome sequencing (WGS) of E. coli is replacing established subtyping methods such as pulsed-field gel electrophoresis, providing a major advancement in the ability to investigate food-borne disease outbreaks and for trace-back to sources. A variety of sequence analysis tools and bioinformatic pipelines are being developed to analyze the vast amount of data generated by WGS and to obtain specific information such as O- and H-group determination and the presence of virulence genes and other genetic markers. PMID:27199968

  16. Advances in molecular serotyping and subtyping of Escherichia coli

    DOE PAGESBeta

    Fratamico, Pina M.; DebRoy, Chitrita; Liu, Yanhong; Needleman, David S.; Baranzoni, Gian Marco; Feng, Peter

    2016-05-03

    Escherichia coli plays an important role as a member of the gut microbiota; however, pathogenic strains also exist, including various diarrheagenic E. coli pathotypes and extraintestinal pathogenic E. coli that cause illness outside of the GI-tract. E. coli have traditionally been serotyped using antisera against the ca. 186 O-antigens and 53 H-flagellar antigens. Phenotypic methods, including bacteriophage typing and O- and H- serotyping for differentiating and characterizing E. coli have been used for many years; however, these methods are generally time consuming and not always accurate. Advances in next generation sequencing technologies have made it possible to develop genetic-based subtypingmore » and molecular serotyping methods for E. coli, which are more discriminatory compared to phenotypic typing methods. Furthermore, whole genome sequencing (WGS) of E. coli is replacing established subtyping methods such as pulsedfield gel electrophoresis, providing a major advancement in the ability to investigate food-borne disease outbreaks and for trace-back to sources. Furthermore, a variety of sequence analysis tools and bioinformatic pipelines are being developed to analyze the vast amount of data generated by WGS and to obtain specific information such as O- and H-group determination and the presence of virulence genes and other genetic markers.« less

  17. A DNA Microarray-Based Assay to Detect Dual Infection with Two Dengue Virus Serotypes

    PubMed Central

    Díaz-Badillo, Alvaro; de Lourdes Muñoz, María; Perez-Ramirez, Gerardo; Altuzar, Victor; Burgueño, Juan; Mendoza-Alvarez, Julio G.; Martínez-Muñoz, Jorge P.; Cisneros, Alejandro; Navarrete-Espinosa, Joel; Sanchez-Sinencio, Feliciano

    2014-01-01

    Here; we have described and tested a microarray based-method for the screening of dengue virus (DENV) serotypes. This DNA microarray assay is specific and sensitive and can detect dual infections with two dengue virus serotypes and single-serotype infections. Other methodologies may underestimate samples containing more than one serotype. This technology can be used to discriminate between the four DENV serotypes. Single-stranded DNA targets were covalently attached to glass slides and hybridised with specific labelled probes. DENV isolates and dengue samples were used to evaluate microarray performance. Our results demonstrate that the probes hybridized specifically to DENV serotypes; with no detection of unspecific signals. This finding provides evidence that specific probes can effectively identify single and double infections in DENV samples. PMID:24776933

  18. Complete genome sequence of Streptococcus mutans GS-5, a serotype c strain.

    PubMed

    Biswas, Saswati; Biswas, Indranil

    2012-09-01

    Streptococcus mutans, a principal causative agent of dental caries, is considered to be the most cariogenic among all oral streptococci. Of the four S. mutans serotypes (c, e, f, and k), serotype c strains predominate in the oral cavity. Here, we present the complete genome sequence of S. mutans GS-5, a serotype c strain originally isolated from human carious lesions, which is extensively used as a laboratory strain worldwide. PMID:22887682

  19. De Novo Amino Acid Biosynthesis Contributes to Salmonella enterica Growth in Alfalfa Seedling Exudates

    PubMed Central

    Kwan, Grace; Pisithkul, Tippapha; Amador-Noguez, Daniel

    2014-01-01

    Salmonella enterica is a member of the plant microbiome. Growth of S. enterica in sprouting-seed exudates is rapid; however, the active metabolic networks essential in this environment are unknown. To examine the metabolic requirements of S. enterica during growth in sprouting-seed exudates, we inoculated alfalfa seeds and identified 305 S. enterica proteins extracted 24 h postinoculation from planktonic cells. Over half the proteins had known metabolic functions, and they are involved in over one-quarter of the known metabolic reactions. Ion and metabolite transport accounted for the majority of detected reactions. Proteins involved in amino acid transport and metabolism were highly represented, suggesting that amino acid metabolic networks may be important for S. enterica growth in association with roots. Amino acid auxotroph growth phenotypes agreed with the proteomic data; auxotrophs in amino acid-biosynthetic pathways that were detected in our screen developed growth defects by 48 h. When the perceived sufficiency of each amino acid was expressed as a ratio of the calculated biomass requirement to the available concentration and compared to growth of each amino acid auxotroph, a correlation between nutrient availability and bacterial growth was found. Furthermore, glutamate transport acted as a fitness factor during S. enterica growth in association with roots. Collectively, these data suggest that S. enterica metabolism is robust in the germinating-alfalfa environment; that single-amino-acid metabolic pathways are important but not essential; and that targeting central metabolic networks, rather than dedicated pathways, may be necessary to achieve dramatic impacts on bacterial growth. PMID:25416761

  20. Streptococcus agalactiae Serotype Distribution and Antimicrobial Susceptibility in Pregnant Women in Gabon, Central Africa.

    PubMed

    Belard, Sabine; Toepfner, Nicole; Capan-Melser, Mesküre; Mombo-Ngoma, Ghyslain; Zoleko-Manego, Rella; Groger, Mirjam; Matsiegui, Pierre-Blaise; Agnandji, Selidji T; Adegnika, Ayôla A; González, Raquel; Kremsner, Peter G; Menendez, Clara; Ramharter, Michael; Berner, Reinhard

    2015-01-01

    Neonatal invasive disease due to Streptococcus agalactiae is life threatening and preventive strategies suitable for resource limited settings are urgently needed. Protective coverage of vaccine candidates based on capsular epitopes will relate to local epidemiology of S. agalactiae serotypes and successful management of critical infections depends on timely therapy with effective antibiotics. This is the first report on serotype distribution and antimicrobial susceptibility of S. agalactiae in pregnant women from a Central African region. Serotypes V, III, and Ib accounted for 88/109 (81%) serotypes and all isolates were susceptible to penicillin and clindamycin while 13% showed intermediate susceptibility to erythromycin. PMID:26603208

  1. Streptococcus agalactiae Serotype Distribution and Antimicrobial Susceptibility in Pregnant Women in Gabon, Central Africa

    PubMed Central

    Belard, Sabine; Toepfner, Nicole; Capan-Melser, Mesküre; Mombo-Ngoma, Ghyslain; Zoleko-Manego, Rella; Groger, Mirjam; Matsiegui, Pierre-Blaise; Agnandji, Selidji T.; Adegnika, Ayôla A.; González, Raquel; Kremsner, Peter G.; Menendez, Clara; Ramharter, Michael; Berner, Reinhard

    2015-01-01

    Neonatal invasive disease due to Streptococcus agalactiae is life threatening and preventive strategies suitable for resource limited settings are urgently needed. Protective coverage of vaccine candidates based on capsular epitopes will relate to local epidemiology of S. agalactiae serotypes and successful management of critical infections depends on timely therapy with effective antibiotics. This is the first report on serotype distribution and antimicrobial susceptibility of S. agalactiae in pregnant women from a Central African region. Serotypes V, III, and Ib accounted for 88/109 (81%) serotypes and all isolates were susceptible to penicillin and clindamycin while 13% showed intermediate susceptibility to erythromycin. PMID:26603208

  2. Development of a TaqMan Array Card for Pneumococcal Serotyping on Isolates and Nasopharyngeal Samples.

    PubMed

    Pholwat, Suporn; Sakai, Fuminori; Turner, Paul; Vidal, Jorge E; Houpt, Eric R

    2016-07-01

    Streptococcus pneumoniae is both a commensal and a major pathogen that causes invasive disease in people of all ages. The introduction of serotype-specific pneumococcal vaccines has reduced the burden of disease but has also led to replacement with new strains; thus, serotyping remains important for vaccine-related disease surveillance. Conventional serotyping methods are laborious and expensive. We developed an easy-to-perform genotypic TaqMan array card (TAC) to identify S. pneumoniae strains, including lytA-based sequences, and 53 sequence-specific PCRs to identify 74 serotypes/serogroups covering all current vaccine types as well as prevalent nonvaccine types. The TAC method was evaluated on 146 clinical S. pneumoniae isolates and 13 nonpneumococcal species that naturally inhabit the upper respiratory tract and yielded 97% (142/146) sensitivity and 100% (13/13) specificity versus results of standard Quellung serotyping. The calculated limit of detection was 20 to 200 fg (∼8 to 84 genome equivalents) per reaction. On 23 blinded nasopharyngeal specimens that were pneumococcus culture positive, the TAC pan-pneumococcus lytA assay was positive in 21 (91% sensitivity versus culture). On TAC lytA-positive specimens, a serotype result was obtained on 86%, and the result was 95% accurate versus the subsequent culture's Quellung result. TAC also detected mixed serotypes in two specimens where Quellung detected only the predominant serotype. This TAC method yields fast and comprehensive serotyping compared to the standard method and may be useful on direct specimens. PMID:27170020

  3. Development of a TaqMan Array Card for Pneumococcal Serotyping on Isolates and Nasopharyngeal Samples

    PubMed Central

    Pholwat, Suporn; Sakai, Fuminori; Turner, Paul; Vidal, Jorge E.

    2016-01-01

    Streptococcus pneumoniae is both a commensal and a major pathogen that causes invasive disease in people of all ages. The introduction of serotype-specific pneumococcal vaccines has reduced the burden of disease but has also led to replacement with new strains; thus, serotyping remains important for vaccine-related disease surveillance. Conventional serotyping methods are laborious and expensive. We developed an easy-to-perform genotypic TaqMan array card (TAC) to identify S. pneumoniae strains, including lytA-based sequences, and 53 sequence-specific PCRs to identify 74 serotypes/serogroups covering all current vaccine types as well as prevalent nonvaccine types. The TAC method was evaluated on 146 clinical S. pneumoniae isolates and 13 nonpneumococcal species that naturally inhabit the upper respiratory tract and yielded 97% (142/146) sensitivity and 100% (13/13) specificity versus results of standard Quellung serotyping. The calculated limit of detection was 20 to 200 fg (∼8 to 84 genome equivalents) per reaction. On 23 blinded nasopharyngeal specimens that were pneumococcus culture positive, the TAC pan-pneumococcus lytA assay was positive in 21 (91% sensitivity versus culture). On TAC lytA-positive specimens, a serotype result was obtained on 86%, and the result was 95% accurate versus the subsequent culture's Quellung result. TAC also detected mixed serotypes in two specimens where Quellung detected only the predominant serotype. This TAC method yields fast and comprehensive serotyping compared to the standard method and may be useful on direct specimens. PMID:27170020

  4. Around the World in 1,475 Salmonella Geo-serotypes

    PubMed Central

    Le Hello, Simon; de Jong, Birgitta; Rolfhamre, Per; Faensen, Daniel; Weill, François-Xavier; Giesecke, Johan

    2016-01-01

    It’s easy to remember Salmonella serotypes names, isn’t it? Surely, this is because the naming system of Salmonella serotypes is by far the most scientist friendly. Traditionally, most Salmonella serotypes have been named after geographic locations. We decided to explore the geographic locations to which Salmonella serotypes refer and describe some unexpected twists in the naming scheme. We found that 93% (n = 1,475) of the 1,585 serotypes could be categorized as geo-serotypes; that is, the name refers to a geographic location. The 3 countries with the most geo-serotypes are Germany, the United Kingdom, and the United States. Other serotype names refer to the name of a person, animal, tribe, or food item or are a composite of symptoms and host. The Salmonella serotypes naming scheme has had a valuable effect on public health microbiology, and in the current era of fast development of whole-genome sequencing, it should remain a reference.

  5. Temporal Variations among Invasive Pneumococcal Disease Serotypes in Children and Adults in Germany (1992–2008)

    PubMed Central

    Imöhl, Matthias; Reinert, Ralf René; van der Linden, Mark

    2010-01-01

    Nationwide surveillance of invasive pneumococcal disease has been conducted in Germany since 1992. From 1992 to 2008, a total of 12,137 isolates from invasive pneumococcal disease were collected. Data on serotypes were available for 9,394 invasive isolates. The leading serotypes were serotypes 14 (16.5%), 3 (8.0%), 7F (7.6%), 1 (7.3%), and 23F (6.0%). Variations in serotype distribution over the years are particularly extensive, especially concerning serotype 14 (min 7.4%, max 33.5%) with the highest percentages among the isolates serotyped from around 1997 to 2006. Serotypes 1 and 7F increased over the last decade. No increase was observed concerning serotype 19A. Higher pneumococcal conjugate vaccine coverages were observed among children (7v, 57.3%; 10v, 72.8%; 13v, 83.5%) than among adults (7v, 39.9%; 10v, 55.5%; 13v, 73.5%). The temporal variations in serotype distribution have to be kept in mind when interpreting vaccine coverages reported in epidemiological studies. PMID:20671944

  6. Dengue virus serotyping based on envelope and membrane and nonstructural protein NS1 serotype-specific capture immunoglobulin M enzyme-linked immunosorbent assays.

    PubMed

    Shu, Pei-Yun; Chen, Li-Kuang; Chang, Shu-Fen; Su, Chien-Ling; Chien, Li-Jung; Chin, Chuan; Lin, Ting-Hsiang; Huang, Jyh-Hsiung

    2004-06-01

    Envelope and membrane (E/M) and nonstructural protein NS1 serotype-specific capture Immunoglobulin M (IgM) enzyme-linked immunosorbent assays (ELISAs) were developed to differentiate four dengue virus serotypes. A total of 93 anti-dengue virus IgM-positive serum samples collected between days 5 and 45 of illness from 59 confirmed dengue patients were analyzed. The results showed that positive serotype specificity could be identified for 86.1 and 47.6% of serum samples tested for E/M-specific IgM antibodies versus 83.3 and 42.9% of serum samples tested for NS1-specific IgM antibodies from patients with primary and secondary dengue virus infections, respectively. Dual analyses with both E/M and NS1 serotype-specific capture IgM ELISAs showed that positive serotype specificity could be correctly identified for 98.6 and 61.9% of all of the primary and secondary serum samples tested, respectively. These findings suggested that E/M and NS1 serotype-specific capture IgM ELISAs have the potential to be of use in dengue virus serotyping. PMID:15184425

  7. Characterization of Small ColE1-Like Plasmids Conferring Kanamycin Resistance in Salmonella enterica subsp. enterica serovars Typhimurium and Newport

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Multi-antibiotic resistant (MR) Salmonella enterica serovars Typhimurium and Newport are an increasing concern in human and animal health. Many strains are known to carry antibiotic resistance determinants on multiple plasmids, yet detailed information is scarce. Three plasmids conferring kanamycin...

  8. Characterization of Salmonella enterica serovar Typhimurium and Salmonella enterica serovar 4,[5],12:i:- isolates from pigs presenting with diarrhea in Korea

    PubMed Central

    LEE, Ki-Eun; LEE, Deog-Yong; CHOI, Hwan-Won; CHAE, Su-Jin; YUN, Young-Sun; LEE, Ki-Chan; CHO, Yun-Sang; YANG, Dong-Kun

    2015-01-01

    Between 2011 and 2012, a total of 896 pig fecal samples were collected from nine provinces in Korea, and 50 salmonella enterica susp. enterica serovar Typhimurium (S. Typhimurium) was isolated. The characteristics of the 50 strains were analyzed, and 4 strains were identified as Salmonella enterica subsp. enterica serovar 4,[5],12:i:-. Salmonella 4,[5],12:i:- could not be distinguished from S. Typhimurium through phage typing, antimicrobial resistance testing or multiple-locus variable-number tandem repeat analysis (MLVA). However, among the four Salmonella 4,[5],12:i:- strains, one (KVCC-BA1400078) was identified as a Salmonella 4,[5],12:i:- clone isolated from humans in the United States, and another (KVCC-BA1400080) was identified as DT193, which has been primarily isolated from humans and animals in European countries. The presence of Salmonella 4,[5],12:i:- in Korea poses a significant threat of horizontal transfer between pigs and humans. PMID:26074410

  9. Gene expression analysis of Salmonella enterica Enteritidis NalR and Salmonella enterica Kentucky 3795 exposed to HCl and acetic acid in rich medium

    Technology Transfer Automated Retrieval System (TEKTRAN)

    In the United States, serovar Kentucky has become one of the most frequently isolated Salmonella enterica serovars from chickens. The reasons for this prevalence are not well understood. Phenotypic comparisons of poultry Salmonella isolates belonging to various serovars demonstrated that serovar Ken...

  10. Cross-sectional study examining Salmonella enterica carriage in subiliac lymph nodes of cull and feedlot cattle at harvest

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Bovine peripheral lymph nodes (LNs), including subiliac LNs, have been identified as a potential source of human exposure to Salmonella enterica, when adipose trim containing these nodes is incorporated into ground beef. In order to gain a better understanding of the burden of S. enterica in periphe...

  11. Characterization of tetracycline resistance in Salmonella enterica strains recovered from irrigation water in the Culiacan Valley, Mexico

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Salmonella enterica is one of the most important pathogens responsible for gastrointestinal infections in humans. The increase of S. enterica strains showing resistance against antibiotics has resulted in limiting the effective treatment of human infections. The present study characterized the resi...

  12. Salmonella enterica serovar Kentucky isolates from dairy cows and poultry demonstrate different evolutionary histories and host-specific polymorphisms

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Salmonella enterica subsp. enterica serovar Kentucky is commonly isolated from dairy cows and poultry in the United States. Although it is not among the most frequently isolated serovars from cases of human salmonellosis, its high prevalence in livestock and poultry indicate it is a potential public...

  13. Cross-sectional study examining Salmonella enterica carriage in subiliac lymph nodes of cull and feedlot cattle at harvest

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Bovine peripheral lymph nodes, including subiliac lymph nodes, have been identified as a potential source of human exposure to Salmonella enterica when trim containing these nodes is incorporated into ground beef. In order to gain a better understanding of the burden of S. enterica in subiliac lymp...

  14. Analysis of antimicrobial resistance genes detected in multidrug-resistant salmonella enterica serovar typhimurium isolated from food animals

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The development of multi drug resistance (MDR) in foodborne pathogens such as Salmonella enterica is a concern for both animal and human health. MDR Salmonella enterica serovar Typhimurium is the most prevalent penta-resistant serovar isolated from animals as part of the National Antimicrobial Resis...

  15. Occurrence of antimicrobial-resistant Escherichia coli and Salmonella enterica in the beef cattle production and processing continuum

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Specific concerns have been raised that 3rd-generation cephalosporin-resistant (3GCr) Escherichia coli, trimethoprim-sulfamethoxazole-resistant (COTr) E. coli, 3GCr Salmonella enterica, and nalidixic acid-resistant (NALr) S. enterica, may be present in cattle production environments, persist through...

  16. Phylogenomic analysis identifies gene gains that define Salmonella enterica subspecies I.

    PubMed

    Lienau, E Kurt; Blazar, Jeffrey M; Wang, Charles; Brown, Eric W; Stones, Robert; Musser, Steven; Allard, Marc W

    2013-01-01

    Comparative methods for analyzing whole genome sequence (WGS) data enable us to assess the genetic information available for reconstructing the evolutionary history of pathogens. We used the comparative approach to determine diagnostic genes for Salmonella enterica subspecies I. S. enterica subsp. I strains are known to infect warm-blooded organisms regularly while its close relatives tend to infect only cold-blooded organisms. We found 71 genes gained by the common ancestor of Salmonella enterica subspecies I and not subsequently lost by any member of this subspecies sequenced to date. These genes included many putative functional phenotypes. Twenty-seven of these genes are found only in Salmonella enterica subspecies I; we designed primers to test these genes for use as diagnostic sequence targets and data mined the NCBI Sequence Read Archive (SRA) database for draft genomes which carried these genes. We found that the sequence specificity and variability of these amplicons can be used to detect and discriminate among 317 different serovars and strains of Salmonella enterica subspecies I. PMID:24204679

  17. Serotype Specific Invasive Capacity and Persistent Reduction in Invasive Pneumococcal Disease

    PubMed Central

    Yildirim, Inci; Hanage, William P.; Lipsitch, Marc; Shea, Kimberly M.; Stevenson, Abbie; Finkelstein, Jonathan; Huang, Susan S.; Lee, Grace M.; Kleinman, Ken; Pelton, SI

    2011-01-01

    Defining the propensity of Streptoccocus pneumoniae (SP) serotypes to invade sterile body sites following nasopharyngeal (NP) acquisition has the potential to inform about how much invasive pneumococcal disease (IPD) may occur in a typical population with a given distribution of carriage serotypes. Data from enhanced surveillance for IPD in Massachusetts children ≤7 years in 2003/04, 2006/07 and 2008/09 seasons and surveillance of SP NP carriage during the corresponding respiratory seasons in 16 Massachusetts communities in 2003/04 and 8 of the 16 communities in both 2006/07 and 2008/09 were used to compute a serotype specific “invasive capacity (IC)” by dividing the incidence of IPD due to serotype x by the carriage prevalence of that same serotype in children of the same age. A total of 206 IPD and 806 NP isolates of SP were collected during the study period. An approximate 50-fold variation in the point estimates between the serotypes having the highest (18C, 33F, 7F, 19A, 3 and 22F) and lowest (6C, 23A, 35F, 11A, 35B, 19F, 15A, and 15BC) IC was observed. Point estimates of IC for most of the common serotypes currently colonizing children in Massachusetts were low and likely explain the continued reduction in IPD from the pre-PCV era in the absence of specific protection against these serotypes. Invasive capacity differs among serotypes and as new pneumococcal conjugate vaccines are introduced, ongoing surveillance will be essential to monitor whether serotypes with high invasive capacity emerge (e.g. 33F, 22F) as successful colonizers resulting in increased IPD incidence due to replacement serotypes. PMID:21029807

  18. The structural characterization of the O-polysaccharide antigen in the lipopolysaccharide of Escherichia coli serotype O118 and its relationship to the O-antigens of Salmonella enterica O47 and Escherichia coli serotype O151

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Mild acid hydrolysis of the lipopolysaccharide produced by Escherichia coli O118:H16 (standard reference strain; NRCC 6613) contained an O-polysaccharide (O-PS) composed of D-galactose, 2-acetimidoylamino-2,6-dideoxy-L-galactose (L-FucNAm), 2-acetamido-2-deoxy-D-glucose, ribitol, and phosphate (1:1...

  19. Efficacy of European starling control to reduce Salmonella enterica contamination in a concentrated animal feeding operation in the Texas panhandle

    PubMed Central

    2011-01-01

    Background European starlings (Sturnus vulgaris) are an invasive bird species known to cause damage to plant and animal agriculture. New evidence suggests starlings may also contribute to the maintenance and spread of diseases within livestock facilities. Identifying and mitigating the risk pathways that contribute to disease in livestock is necessary to reduce production losses and contamination of human food products. To better understand the impact starlings have on disease transmission to cattle we assessed the efficacy of starling control as a tool to reduce Salmonella enterica within a concentrated animal feeding operation. We matched a large facility, slated for operational control using DRC-1339 (3-chloro-4-methylaniline hydrochloride, also 3-chloro p-toluidine hydrochloride, 3-chloro-4-methylaniline), with a comparable reference facility that was not controlling birds. In both facilities, we sampled cattle feed, cattle water and cattle feces for S. enterica before and after starling control operations. Results Within the starling-controlled CAFO, detections of S. enterica contamination disappeared from feed bunks and substantially declined within water troughs following starling control operations. Within the reference facility, detections of S. enterica contamination increased substantially within feed bunks and water troughs. Starling control was not observed to reduce prevalence of S. enterica in the cattle herd. Following starling control operations, herd prevalence of S. enterica increased on the reference facility but herd prevalence of S. enterica on the starling-controlled CAFO stayed at pretreatment levels. Conclusions Within the starling-controlled facility detections of S. enterica disappeared from feed bunks and substantially declined within water troughs following control operations. Since cattle feed and water are obvious routes for the ingestion of S. enterica, starling control shows promise as a tool to help livestock producers manage

  20. Novel strain of Shigella dysenteriae serotype 7 from India

    PubMed Central

    Mandal, J.; Poonambath, D.K.; Bhosale, N.K.; Das, A.

    2015-01-01

    We describe a strain of Shigella dysenteriae serotype 7 which had novel biochemical and genetic characters. Unlike other S. dysenteriae, it produced gas, fermented mannitol, was a late-lactose fermenter and harboured the set 1A and set 1B genes. The significance of such atypical strains is that they are difficult to identify. If such strains are missed, they could prove to be a serious public health problem because the infectious dose is very low and they may harbour integrons contributing to drug resistance. PMID:26442152

  1. Plane of nutrition influences the performance, innate leukocyte responses, and resistance to an oral Salmonella enterica serotype Typhimurium challenge in Jersey calves

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Two experiments were conducted to determine the influence of plane of nutrition on performance, leukocyte responses, and the pathophysiological response to an oral Salmonella typhimurium challenge in Jersey calves. In experiment 1, forty-six (2 ± 1 days of age) calves were randomly assigned to 2 die...

  2. Part I. Analysis of data gaps pertaining to Salmonella enterica serotype Typhi infections in low and medium human development index countries, 1984–2005

    PubMed Central

    CRUMP, J. A.; RAM, P. K.; GUPTA, S. K.; MILLER, M. A.; MINTZ, E. D.

    2008-01-01

    SUMMARY There are only 10 contemporary, population-based studies of typhoid fever that evaluate disease incidence using blood culture for confirmation of cases. Reported incidence ranged from 13 to 976/100 000 persons per year. These studies are likely to have been done preferentially in high- incidence sites which makes generalization of data difficult. Only five of these studies reported mortality. Of these the median (range) mortality was 0% (0–1·8%). Since study conditions usually involved enhanced clinical management of patients and the studies were not designed to evaluate mortality as an outcome, their usefulness for generalizing case-fatality rates is uncertain. No contemporary population-based studies reported rates of complications. Hospital-based typhoid fever studies reported median (range) complication rates of 2·8% (0·6–4·9%) for intestinal perforation and case-fatality rates of 2·0% (0–14·8%). Rates of complications other than intestinal perforation were not reported in contemporary hospital-based studies. Hospital-based studies capture information on the most severe illnesses among persons who have access to health-care services limiting their generalizability. Only two studies have informed the current understanding of typhoid fever age distribution curves. Extrapolation from population-based studies suggests that most typhoid fever occurs among young children in Asia. To reduce gaps in the current understanding of typhoid fever incidence, complications, and case-fatality rate, large population-based studies using blood culture confirmation of cases are needed in representative sites, especially in low and medium human development index countries outside Asia. PMID:17686194

  3. Contribution of the Type VI Secretion System Encoded in SPI-19 to Chicken Colonization by Salmonella enterica Serotypes Gallinarum and Enteritidis

    PubMed Central

    Blondel, Carlos J.; Yang, Hee-Jeong; Castro, Benjamín; Chiang, Sebastián; Toro, Cecilia S.; Zaldívar, Mercedes; Contreras, Inés; Andrews-Polymenis, Helene L.; Santiviago, Carlos A.

    2010-01-01

    Salmonella Gallinarum is a pathogen with a host range specific to poultry, while Salmonella Enteritidis is a broad host range pathogen that colonizes poultry sub-clinically but is a leading cause of gastrointestinal salmonellosis in humans and many other species. Despite recent advances in our understanding of the complex interplay between Salmonella and their hosts, the molecular basis of host range restriction and unique pathobiology of Gallinarum remain largely unknown. Type VI Secretion System (T6SS) represents a new paradigm of protein secretion that is critical for the pathogenesis of many Gram-negative bacteria. We recently identified a putative T6SS in the Salmonella Pathogenicity Island 19 (SPI-19) of Gallinarum. In Enteritidis, SPI-19 is a degenerate element that has lost most of the T6SS functions encoded in the island. In this work, we studied the contribution of SPI-19 to the colonization of Salmonella Gallinarum strain 287/91 in chickens. Non-polar deletion mutants of SPI-19 and the clpV gene, an essential T6SS component, colonized the ileum, ceca, liver and spleen of White Leghorn chicks poorly compared to the wild-type strain after oral inoculation. Return of SPI-19 to the ΔSPI-19 mutant, using VEX-Capture, complemented this colonization defect. In contrast, transfer of SPI-19 from Gallinarum to Enteritidis resulted in transient increase in the colonization of the ileum, liver and spleen at day 1 post-infection, but at days 3 and 5 post-infection a strong colonization defect of the gut and internal organs of the experimentally infected chickens was observed. Our data indicate that SPI-19 and the T6SS encoded in this region contribute to the colonization of the gastrointestinal tract and internal organs of chickens by Salmonella Gallinarum and suggest that degradation of SPI-19 T6SS in Salmonella Enteritidis conferred an advantage in colonization of the avian host. PMID:20661437

  4. Fibronectin Binding to the Salmonella enterica Serotype Typhimurium ShdA Autotransporter Protein Is Inhibited by a Monoclonal Antibody Recognizing the A3 Repeat

    PubMed Central

    Kingsley, Robert A.; Abi Ghanem, Daad; Puebla-Osorio, Nahum; Keestra, A. Marijke; Berghman, Luc; Bäumler, Andreas J.

    2004-01-01

    ShdA is a large outer membrane protein of the autotransporter family whose passenger domain binds the extracellular matrix proteins fibronectin and collagen I, possibly by mimicking the host ligand heparin. The ShdA passenger domain consists of ∼1,500 amino acid residues that can be divided into two regions based on features of the primary amino acid sequence: an N-terminal nonrepeat region followed by a repeat region composed of two types of imperfect direct amino acid repeats, called type A and type B. The repeat region bound bovine fibronectin with an affinity similar to that for the complete ShdA passenger domain, while the nonrepeat region exhibited comparatively low fibronectin-binding activity. A number of fusion proteins containing truncated fragments of the repeat region did not bind bovine fibronectin. However, binding of the passenger domain to fibronectin was inhibited in the presence of immune serum raised to one truncated fragment of the repeat region that contained repeats A2, B8, A3, and B9. Furthermore, a monoclonal antibody that specifically recognized an epitope in a recombinant protein containing the A3 repeat inhibited binding of ShdA to fibronectin. PMID:15262930

  5. Fibronectin binding to the Salmonella enterica serotype Typhimurium ShdA autotransporter protein is inhibited by a monoclonal antibody recognizing the A3 repeat.

    PubMed

    Kingsley, Robert A; Abi Ghanem, Daad; Puebla-Osorio, Nahum; Keestra, A Marijke; Berghman, Luc; Bäumler, Andreas J

    2004-08-01

    ShdA is a large outer membrane protein of the autotransporter family whose passenger domain binds the extracellular matrix proteins fibronectin and collagen I, possibly by mimicking the host ligand heparin. The ShdA passenger domain consists of approximately 1,500 amino acid residues that can be divided into two regions based on features of the primary amino acid sequence: an N-terminal nonrepeat region followed by a repeat region composed of two types of imperfect direct amino acid repeats, called type A and type B. The repeat region bound bovine fibronectin with an affinity similar to that for the complete ShdA passenger domain, while the nonrepeat region exhibited comparatively low fibronectin-binding activity. A number of fusion proteins containing truncated fragments of the repeat region did not bind bovine fibronectin. However, binding of the passenger domain to fibronectin was inhibited in the presence of immune serum raised to one truncated fragment of the repeat region that contained repeats A2, B8, A3, and B9. Furthermore, a monoclonal antibody that specifically recognized an epitope in a recombinant protein containing the A3 repeat inhibited binding of ShdA to fibronectin. PMID:15262930

  6. Prevalence, enumeration, serotypes, and antimicrobial resistance phenotypes of Salmonella enterica isolates from carcasses at two large United States pork processing plants

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The objective of this study was to characterize Salmonella contamination on carcasses in two large U.S. commercial pork processing plants. Carcasses were sampled before scalding, after dehairing/polishing but before evisceration, and after chilling on two days in each of the four seasons. The prev...

  7. Generation of serotype 1/serotype 2 reassortant viruses of the infectious bursal disease virus and their investigation in vitro and in vivo.

    PubMed

    Zierenberg, Kati; Raue, Rüdiger; Nieper, Hermann; Islam, Md Rafiqul; Eterradossi, Nicolas; Toquin, Didier; Müller, Hermann

    2004-09-15

    Infectious bursal disease virus (IBDV) is the causative agent of acute or immunosuppressive disease in chickens. Serotype 1 strains are pathogenic whereas serotype 2 strains neither cause disease nor protect against infection with the serotype 1 strains. The target organ of serotype 1 strains is the bursa Fabricii (BF). The molecular determinants of this tropism, and therefore pathogenicity, are poorly understood. IBDV is a non-enveloped icosahedral virus particle of 60 nm in diameter, which contains two genome segments of double-stranded RNA. Here, the generation of interserotypic reassortants using the reverse genetics approach is reported. The results of in vitro and in vivo investigations show that genome segment A determines the bursa tropism of IBDV, whereas segment B is involved in the efficiency of viral replication; they further indicate the significance of the interaction of the polymerase (segment B) with the structural protein VP3 (segment A) or the viral genome for efficient virus formation and replication. PMID:15325078

  8. Draft Genome Sequences of Streptococcus agalactiae Serotype Ia and III Isolates from Tilapia Farms in Thailand.

    PubMed

    Areechon, Nontawith; Kannika, Korntip; Hirono, Ikuo; Kondo, Hidehiro; Unajak, Sasimanas

    2016-01-01

    Streptococcus agalactiaeserotypes Ia and III were isolated from infected tilapia in cage and pond culture farms in Thailand during 2012 to 2014, in which pathogenicity analysis demonstrated that serotype III showed higher virulence than serotype Ia. Here, we report the draft genome sequencing of piscineS. agalactiaeserotypes Ia and III. PMID:27013037

  9. Clinical Implications of Pneumococcal Serotypes: Invasive Disease Potential, Clinical Presentations, and Antibiotic Resistance

    PubMed Central

    Nahm, Moon H.; Moseley, M. Allen

    2013-01-01

    Streptococcus pneumoniae can asymptomatically colonize the nasopharynx and cause a diverse range of illnesses. This clinical spectrum from colonization to invasive pneumococcal disease (IPD) appears to depend on the pneumococcal capsular serotype rather than the genetic background. According to a literature review, serotypes 1, 4, 5, 7F, 8, 12F, 14, 18C, and 19A are more likely to cause IPD. Although serotypes 1 and 19A are the predominant causes of invasive pneumococcal pneumonia, serotype 14 remains one of the most common etiologic agents of non-bacteremic pneumonia in adults, even after 7-valent pneumococcal conjugate vaccine (PCV7) introduction. Serotypes 1, 3, and 19A pneumococci are likely to cause empyema and hemolytic uremic syndrome. Serotype 1 pneumococcal meningitis is prevalent in the African meningitis belt, with a high fatality rate. In contrast to the capsule type, genotype is more closely associated with antibiotic resistance. CC320/271 strains expressing serotype 19A are multidrug-resistant (MDR) and prevalent worldwide in the era of PCV7. Several clones of MDR serotype 6C pneumococci emerged, and a MDR 6D clone (ST282) has been identified in Korea. Since the pneumococcal epidemiology of capsule types varies geographically and temporally, a nationwide serosurveillance system is vital to establishing appropriate vaccination strategies for each country. PMID:23341706

  10. Changes in Streptococcus pneumoniae Serotype 19A Invasive Infections in Children from 1993 to 2011

    PubMed Central

    Kaplan, Sheldon L.; Lamberth, Linda B.; Barson, William J.; Romero, José R.; Lin, Philana Ling; Bradley, John S.; Givner, Laurence B.; Tan, Tina Q.; Hoffman, Jill A.; Mason, Edward O.

    2013-01-01

    Among 594 Streptococcus pneumoniae serotype 19A invasive pneumococcal disease (IPD) isolates collected from 1993 to 2011, we identified 85 sequence types by multilocus sequence typing. CC320 was associated with multidrug resistance and reduced susceptibility to penicillin and ceftriaxone and still predominated among declining serotype 19A IPD isolates following PCV13 introduction. PMID:23390277

  11. A molecular scheme for Yersinia enterocolitica patho-serotyping derived from genome-wide analysis.

    PubMed

    Garzetti, Debora; Susen, Rosa; Fruth, Angelika; Tietze, Erhard; Heesemann, Jürgen; Rakin, Alexander

    2014-05-01

    Yersinia enterocolitica is a food-borne, gastro-intestinal pathogen with world-wide distribution. Only 11 serotypes have been isolated from patients, with O:3, O:9, O:8 and O:5,27 being the serotypes most commonly associated with human yersiniosis. Serotype is an important characteristic of Y. enterocolitica strains, allowing differentiation for epidemiology, diagnosis and phylogeny studies. Conventional serotyping, performed by slide agglutination, is a tedious and laborious procedure whose interpretation tends to be subjective, leading to poor reproducibility. Here we present a PCR-based typing scheme for molecular identification and patho-serotyping of Y. enterocolitica. Genome-wide comparison of Y. enterocolitica sequences allowed analysis of the O-antigen gene clusters of different serotypes, uncovering their formerly unknown genomic locations, and selection of targets for serotype-specific amplification. Two multiplex PCRs and one additional PCR were designed and tested on various reference strains and isolates from different origins. Our genotypic assay proved to be highly specific for identification of Y. enterocolitica species, discrimination between virulent and non-virulent strains, distinguishing the main human-related serotypes, and typing of conventionally untypeable strains. This genotyping scheme could be applied in microbiology laboratories as an alternative or complementary method to the traditional phenotypic assays, providing data for epidemiological studies. PMID:24246413

  12. Serotyping of Streptococcus pneumoniae by agglutination assays: a cost-effective technique for developing countries.

    PubMed Central

    Lalitha, M. K.; Pai, R.; John, T. J.; Thomas, K.; Jesudason, M. V.; Brahmadathan, K. N.; Sridharan, G.; Steinhoff, M. C.

    1996-01-01

    There is a need for additional data on the distribution of pneumococcal serotypes in developing countries. We report the use of a coagglutination (COA) and a latex agglutination (LA) test for serotyping Streptococcus pneumoniae which were evaluated using 114 clinical isolates in Vellore, India. In tests to serotype 30 fresh isolates of pneumococci from meningitis (8 isolates), bacteraemia/septicaemia (21 isolates) and peritonitis (1 isolate) cases, there was complete concordance among the three methods. An additional 20 isolates (11 from cerebrospinal fluid and 9 from blood cultures) were serotyped using both LA and COA, with full agreement between the results. With a further 30 isolates, there was 93% concordance for the COA types with serotypes assigned by a WHO reference laboratory. The COA and LA serotyping results were equivalent in accuracy to those obtained using quellung serotyping. Both these agglutination tests are rapid, valid, and relatively cheap, and with appropriate validation by reference laboratories they could be more widely used in developing countries to obtain local and regional data on pneumococcal serotype distribution. PMID:8823960

  13. A 7-plex microbead-based immunoassay for serotyping Shiga toxin-producing Escherichia coli

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Serotyping of Shiga toxin-producing Escherichia coli (STEC) has been contingent upon the availability of antisera. Here we describe a 7-plex microbead-based immunoassay to simultaneously serotype seven STEC (i.e., belonging to serogroups O26, O45, O103, O111, O121, O145, and O157) by the Luminex xMA...

  14. Complete Genome Sequence of Aggregatibacter actinomycetemcomitans Serotype g Strain NUM4039 (JCM 30399)

    PubMed Central

    Saito, Masanori; Hirasawa, Masaaki; Kuwahara, Noriko; Okada, Tamami; Umezawa, Koji; Kobayashi, Taira; Okamoto, Masaaki; Naito, Mariko; Hirasawa, Masatomo

    2016-01-01

    Aggregatibacter actinomycetemcomitans is considered to be a major etiological agent of aggressive periodontitis and includes serotype a to g strains. We herein report the first complete genome sequence of A. actinomycetemcomitans serotype g strain NUM4039. The genome is 2,382,853 bp in length with a G+C content of 44.34%. PMID:26988057

  15. Determination of the median lethal dose of botulinum serotype E in channel catfish fingerlings

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The median lethal dose of botulinum serotype E in 5.3-g channel catfish Ictalurus punctatus fingerlings was determined. Five tanks (five fish/tank) were assigned to each of the following treatment groups: 70, 50, 35, 25, or 15 pg of purified botulinum serotype E. Fish were injected intracoelomically...

  16. A monoclonal antibody based capture ELISA for botulinum neurotoxin serotype B: toxin detection in food

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Botulism is a serious foodborne neuroparalyic disease caused by botulinum neurotoxin (BoNT) produced by the anaerobic bacterium Clostridium botulinum. Seven toxin serotypes (A-H) have been described. The majority of human cases of botulism are caused by serotypes A and B followed by E and F. We repo...

  17. Single-Step Multiplex PCR Assay for Determining 92 Pneumococcal Serotypes.

    PubMed

    Marimón, José M; Ercibengoa, María; Santacatterina, Erica; Alonso, Marta; Pérez-Trallero, Emilio

    2016-08-01

    For pneumococcal disease surveillance, simple and cost-effective methods capable of determining all serotypes are needed. Combining a single-tube multiplex PCR with fluorescently labeled primers followed by amplicon analysis using automated fluorescent capillary electrophoresis, each serotype of 92 reference isolates and 297 recently collected clinical isolates was successfully determined. PMID:27280423

  18. Susceptibility of North American white-tailed deer to the Netherlands strain of BTV serotype 8

    Technology Transfer Automated Retrieval System (TEKTRAN)

    World-wide there are at least 24 serotypes of bluetongue virus (BTV), a complex non-enveloped virus in the family Reoviridae, genus Orbivirus. Bluetongue (BT) is an arthropod-borne disease of cattle, sheep, goats, and deer and is transmitted by Culicoides biting midges. In 2006, bluetongue serotype ...

  19. Complete Coding Genome Sequence of Putative Novel Bluetongue Virus Serotype 27

    PubMed Central

    Jenckel, Maria; Bréard, Emmanuel; Schulz, Claudia; Sailleau, Corinne; Viarouge, Cyril; Hoffmann, Bernd; Beer, Martin; Zientara, Stéphan

    2015-01-01

    We announce the complete coding genome sequence of a novel bluetongue virus (BTV) serotype (BTV-n = putative BTV-27) detected in goats in Corsica, France, in 2014. Sequence analysis confirmed the closest relationship between sequences of the novel BTV serotype and BTV-25 and BTV-26, recently discovered in Switzerland and Kuwait, respectively. PMID:25767218

  20. Experimental infection of white-tailed deer (Odocoileus virginianus) with Northern European bluetongue virus serotype 8

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Bluetongue (BT) is an insect-transmitted, economically important disease of domestic and wild ruminants. Although only five of the 26 reported bluetongue virus (BTV) serotypes are considered endemic to the USA, 10 exotic serotypes have been isolated primarily in the southeastern region of the count...

  1. Emergence of a new multidrug-resistant serotype X variant in an epidemic clone of Shigella flexneri.

    PubMed

    Ye, Changyun; Lan, Ruiting; Xia, Shengli; Zhang, Jin; Sun, Qiangzheng; Zhang, Shaomin; Jing, Huaiqi; Wang, Lei; Li, Zhenjun; Zhou, Zhemin; Zhao, Ailan; Cui, Zhigang; Cao, Jingjing; Jin, Dong; Huang, Lili; Wang, Yiting; Luo, Xia; Bai, Xuemei; Wang, Yan; Wang, Ping; Xu, Qiang; Xu, Jianguo

    2010-02-01

    Shigella spp. are the causative agent of shigellosis with Shigella flexneri serotype 2a being the most prevalent in developing countries. Epidemiological surveillance in China found that a new serotype of S. flexneri appeared in 2001 and replaced serotype 2a in 2003 as the most prevalent serotype in Henan Province. The new serotype also became the dominant serotype in 7 of the 10 other provinces under surveillance in China by 2007. The serotype was identified as a variant of serotype X. It differs from serotype X by agglutination to the monovalent anti-IV type antiserum and the group antigen-specific monoclonal antibody MASF IV-I. Genome sequencing of a serotype X variant isolate, 2002017, showed that it acquired a Shigella serotype conversion island, also as an SfX bacteriophage, containing gtr genes for type X-specific glucosylation. Multilocus sequence typing of 15 genes from 37 serotype X variant isolates and 69 isolates of eight other serotypes, 1a, 2a, 2b, 3a, 4a, 5b, X, and Y, found that all belong to a new sequence type (ST), ST91. Pulsed-field gel electrophoresis revealed 154 pulse types with 655 S. flexneri isolates analyzed and identified 57 serotype switching events. The data suggest that S. flexneri epidemics in China have been caused by a single epidemic clone, ST91, with frequent serotype switching to evade infection-induced immunity to serotypes to which the population was exposed previously. The clone has also acquired resistance to multiple antibiotics. These findings underscore the challenges to the current vaccine development and control strategies for shigellosis. PMID:19955273

  2. The prevalence of Salmonella enterica in Spanish feed mills and potential feed-related risk factors for contamination.

    PubMed

    Torres, Gregorio J; Piquer, F Javier; Algarra, Leonor; de Frutos, Cristina; Sobrino, Odón J

    2011-02-01

    A cross-sectional study was conducted in Spain to estimate the prevalence of Salmonella enterica in feed mills and to identify and evaluate potential risk factors associated with feed contamination. A total of 3844 samples were collected from 523 different feed mills using a stratified sampling method. Samples were tested for the presence of Salmonella using conventional culture methods. When the presence of Salmonella was detected, samples were further characterised using serotyping at the National Reference Laboratory (NRL) for animal feed. Additional data about the biosecurity and hygiene measures, feed material used and compound feed produced, were collected by official veterinarians using a questionnaire in situ. In 144 of the feed mills visited (28%), Salmonella were present. However, it was only isolated from 4.8% of samples taken from all of the feed mills (3.5% from feed materials, 3.2% from compound feed and 12.5% from dust of the feed mill facilities). Salmonella serovars of public health importance (Enteritidis, Typhimurium, Infantis, Virchow and Hadar), were detected in only 2.7% of feed mills and in 0.3% of the samples studied. Logistic regression was used to investigate potential feed-mill risk factors for the isolation of Salmonella. Feed mill intake pits were demonstrated to have an increased risk of culture-positive dust samples (OR=6.4; 95% CI: 2.7-15.1). The feed material used in the production of compound feed was associated with recovery of Salmonella. Of the feed material used, cotton seeds were identified as having the highest odds of contamination (OR=3.8; 95% CI: 1.7-8.3). Pelleting appears to reduce the chance of contamination because non-pelleted compound feed is 8 times more likely to be contaminated than pelleted compound feed (OR=8.2; 95% CI: 2.5-26.6). The role of the feed itself in the epidemiology of Salmonella seems to be of limited importance as compound feed is not frequently contaminated at the feed mill level. This should not

  3. Antimicrobial Resistance and Molecular Investigation of H2S-Negative Salmonella enterica subsp. enterica serovar Choleraesuis Isolates in China

    PubMed Central

    Li, Peng; Liang, Beibei; Li, Hao; Yang, Xiaoxia; Wang, Ligui; Hao, Rongzhang; Jia, Leili; Wu, Zhihao; Qiu, Shaofu; Song, Hongbin

    2015-01-01

    Salmonella enterica subsp. enterica serovar Choleraesuis is a highly invasive pathogen of swine that frequently causes serious outbreaks, in particular in Asia, and can also cause severe invasive disease in humans. In this study, 21 S. Choleraesuis isolates, detected from 21 patients with diarrhea in China between 2010 and 2011, were found to include 19 H2S-negative S. Choleraesuis isolates and two H2S-positive isolates. This is the first report of H2S-negative S. Choleraesuis isolated from humans. The majority of H2S-negative isolates exhibited high resistance to ampicillin, chloramphenicol, gentamicin, tetracycline, ticarcillin, and trimethoprim-sulfamethoxazole, but only six isolates were resistant to norfloxacin. In contrast, all of the isolates were sensitive to cephalosporins. Fifteen isolates were found to be multidrug resistant. In norfloxacin-resistant isolates, we detected mutations in the gyrA and parC genes and identified two new mutations in the parC gene. Pulsed-field gel electrophoresis (PFGE), multilocus sequence typing (MLST), and clustered regularly interspaced short palindromic repeat (CRISPR) analysis were employed to investigate the genetic relatedness of H2S-negative and H2S-positive S. Choleraesuis isolates. PFGE revealed two groups, with all 19 H2S-negative S. Choleraesuis isolates belonging to Group I and H2S-positive isolates belonging to Group II. By MLST analysis, the H2S-negative isolates were all found to belong to ST68 and H2S-positive isolates belong to ST145. By CRISPR analysis, no significant differences in CRISPR 1 were detected; however, one H2S-negative isolate was found to contain three new spacers in CRISPR 2. All 19 H2S-negative isolates also possessed a frame-shift mutation at position 760 of phsA gene compared with H2S-positive isolates, which may be responsible for the H2S-negative phenotype. Moreover, the 19 H2S-negative isolates have similar PFGE patterns and same mutation site in the phsA gene, these results indicated

  4. Evaluation of Two Enzyme-Linked Immunosorbent Assays for Detecting Salmonella enterica subsp. enterica Serovar Dublin Antibodies in Bulk Milk

    PubMed Central

    Veling, J.; van Zijderveld, F. G.; van Zijderveld-van Bemmel, A. M.; Schukken, Y. H.; Barkema, H. W.

    2001-01-01

    Two enzyme-linked immunosorbent assays (ELISAs) for the detecting Salmonella enterica subsp. enterica serovar Dublin antibodies in bulk milk were developed and evaluated for potential use in control programs. The ELISAs were based on either lipopolysacharide (LPS ELISA) or flagellar antigen (GP ELISA). Sensitivity was determined with 79 case herds with a wide range of clinical signs. Specificity was determined with 125 Dutch and 200 Swedish control herds. The relation between antibodies in bulk milk, antibodies in serum, and the level of milk production of individual cows was studied with 61 case herds. The optimal optical density (OD) values of the LPS ELISA and the GP ELISA were determined to be 0.2 and 0.5, respectively. The sensitivities of the LPS ELISA and the GP ELISA were 54 and 63%, respectively, with a specificity of 98% for both ELISAs with samples from the Dutch control herds. The specificities for samples from the Swedish herds were 100% for the LPS ELISA and 95% for the GP ELISA. The sensitivity of the combination of tests was 65% when samples were run in parallel, and the specificity was 100% when samples were run in series, irrespective of whether the samples came from Dutch or Swedish control herds. The variance (R2) in the OD value for bulk milk samples could be explained by the percentage of seropositive lactating cows in a herd with the LPS ELISA for 51% of the samples and with the GP ELISA for 72%. The variance in the OD value was best explained by the combination of the percentage of seropositive lactating cows in the herd and the mean log10 serum antibody titer for that herd (R2 = 62% for the LPS ELISA and R2 = 75% for the GP ELISA). Case herds more often tested negative by the ELISA with bulk milk when the percentage of seropositive lactating cows was less than 5%. It is concluded that both ELISAs with bulk milk can be used in control programs to distinguish between infected and noninfected herds. Specificity can be increased by using the

  5. The serological relationship between Brucella spp., Yersinia enterocolitica serotype IX and Salmonella serotypes of Kauffmann-White group N.

    PubMed Central

    Corbell, M. J.

    1975-01-01

    The serological relationship between Brucella spp., Yersinia enterocolitica IX, and the group N salmonella serotypes S. godesberg, S. landau, S. morehead, S. neusdorf, S. soerenga and S. urbana was examined using agglutination, antiglobulin, complement fixation, immunodiffusion and fluorescent antibody methods. Antisera to the group N salmonella serotypes all reacted to significant titres in agglutination and complement fixation, but not antiglobulin or immunodiffusion tests with smooth brucella antigens. These antisera also reacted in agglutination, but not antiglobulin, tests with Y. enterocolitica IX. They did not react significantly in any tests with rough brucella antigens. Conversely, antisera to smooth Brucella spp. agglutinated group N salmonellas to low titre and Y. enterocolitica IX to titres similar to those given against the homologous strain. Antiserum to Y. enterocolitica IX on the other hand reacted with smooth brucella antigens to high titre in agglutination, complement fixation and antiglobulin tests, and with the group N salmonella antigens to substantial titres in agglutination tests. In direct fluorescent antibody tests, smooth Brucella strains and Y. enterocolitica IX reacted strongly with FITC-labelled antibody to Br. abortus whereas the group N salmonella strains reacted weakly. In tests with monospecific antisera to the A and M determinants of Br. abortus and Br. melitensis respectively, Y. enterocolitica IX reacted only with the antiserum to the A determinant whereas group N salmonellas reacted to low titre with both A and M antisera. The results of cross-absorption tests confirmed this relationship and suggested that the O30 antigens of group N salmonella serotypes contained antigenic determinants similar to, but not identical with, the antigenic structure shared by smooth Brucella spp. and Y. enterocolitica IX. PMID:807618

  6. The giant adhesin SiiE of Salmonella enterica.

    PubMed

    Barlag, Britta; Hensel, Michael

    2015-01-01

    Salmonella enterica is a Gram-negative, food-borne pathogen, which colonizes the intestinal tract and invades enterocytes. Invasion of polarized cells depends on the SPI1-encoded type III secretion system (T3SS) and the SPI4-encoded type I secretion system (T1SS). The substrate of this T1SS is the non-fimbrial giant adhesin SiiE. With a size of 595 kDa, SiiE is the largest protein of the Salmonella proteome and consists of 53 repetitive bacterial immunoglobulin (BIg) domains, each containing several conserved residues. As known for other T1SS substrates, such as E. coli HlyA, Ca2+ ions bound by conserved D residues within the BIg domains stabilize the protein and facilitate secretion. The adhesin SiiE mediates the first contact to the host cell and thereby positions the SPI1-T3SS to initiate the translocation of a cocktail of effector proteins. This leads to actin remodeling, membrane ruffle formation and bacterial internalization. SiiE binds to host cell apical membranes in a lectin-like manner. GlcNAc and α2-3 linked sialic acid-containing structures are ligands of SiiE. Since SiiE shows repetitive domain architecture, we propose a zipper-like binding mediated by each individual BIg domain. In this review, we discuss the characteristics of the SPI4-T1SS and the giant adhesin SiiE. PMID:25587788

  7. Clustered Intracellular Salmonella enterica Serovar Typhimurium Blocks Host Cell Cytokinesis

    PubMed Central

    Durkin, Charlotte H.; Helaine, Sophie; Boucrot, Emmanuel

    2016-01-01

    Several bacterial pathogens and viruses interfere with the cell cycle of their host cells to enhance virulence. This is especially apparent in bacteria that colonize the gut epithelium, where inhibition of the cell cycle of infected cells enhances the intestinal colonization. We found that intracellular Salmonella enterica serovar Typhimurium induced the binucleation of a large proportion of epithelial cells by 14 h postinvasion and that the effect was dependent on an intact Salmonella pathogenicity island 2 (SPI-2) type 3 secretion system. The SPI-2 effectors SseF and SseG were required to induce binucleation. SseF and SseG are known to maintain microcolonies of Salmonella-containing vacuoles close to the microtubule organizing center of infected epithelial cells. During host cell division, these clustered microcolonies prevented the correct localization of members of the chromosomal passenger complex and mitotic kinesin-like protein 1 and consequently prevented cytokinesis. Tetraploidy, arising from a cytokinesis defect, is known to have a deleterious effect on subsequent cell divisions, resulting in either chromosomal instabilities or cell cycle arrest. In infected mice, proliferation of small intestinal epithelial cells was compromised in an SseF/SseG-dependent manner, suggesting that cytokinesis failure caused by S. Typhimurium delays epithelial cell turnover in the intestine. PMID:27185791

  8. Salmonella enterica induces and subverts the plant immune system

    PubMed Central

    García, Ana V.; Hirt, Heribert

    2014-01-01

    Infections with Salmonella enterica belong to the most prominent causes of food poisoning and infected fruits and vegetables represent important vectors for salmonellosis. Although it was shown that plants raise defense responses against Salmonella, these bacteria persist and proliferate in various plant tissues. Recent reports shed light into the molecular interaction between plants and Salmonella, highlighting the defense pathways induced and the means used by the bacteria to escape the plant immune system and accomplish colonization. It was recently shown that plants detect Salmonella pathogen-associated molecular patterns (PAMPs), such as the flagellin peptide flg22, and activate hallmarks of the defense program known as PAMP-triggered immunity (PTI). Interestingly, certain Salmonella strains carry mutations in the flg22 domain triggering PTI, suggesting that a strategy of Salmonella is to escape plant detection by mutating PAMP motifs. Another strategy may rely on the type III secretion system (T3SS) as T3SS mutants were found to induce stronger plant defense responses than wild type bacteria. Although Salmonella effector delivery into plant cells has not been shown, expression of Salmonella effectors in plant tissues shows that these bacteria also possess powerful means to manipulate the plant immune system. Altogether, these data suggest that Salmonella triggers PTI in plants and evolved strategies to avoid or subvert plant immunity. PMID:24772109

  9. Simultaneous Rapid Detection and Serotyping of Cronobacter sakazakii Serotypes O1, O2, and O3 by Using Specific Monoclonal Antibodies.

    PubMed

    Scharinger, Eva J; Dietrich, Richard; Kleinsteuber, Ina; Märtlbauer, Erwin; Schauer, Kristina

    2016-04-01

    Cronobacter sakazakii is a foodborne pathogen associated with rare but often lethal infections in neonates. Powdered infant formula (PIF) represents the most frequent source of infection. Out of the identified serotypes (O1 to O7), O1, O2, and O3 are often isolated from clinical and PIF samples. Serotype-specific monoclonal antibodies (MAbs) suitable for application in enzyme immunoassays (EIAs) for the rapid detection of C. sakazakii have not yet been developed. In this study, we created specific MAbs with the ability to bind toC. sakazakii of serotypes O1, O2, and O3. Characterization by indirect EIAs, immunofluorescence, motility assays, and immunoblotting identified lipopolysaccharide (LPS) and exopolysaccharide (EPS) as the antigenic determinants of the MAbs. The established sandwich EIAs were highly sensitive and were able to detect between 2 × 10(3)and 9 × 10(6)CFU/ml. Inclusivity tests confirmed that 93% of serotype O1 strains, 100% of O2 strains, and 87% of O3 strains were detected at low cell counts. No cross-reactivity with >100 strains of Cronobacter spp. and other Enterobacter iaceae was observed, except for that with C. sakazakii serotype O3 and Cronobacter muytjensii serotype O1. Moreover, the sandwich EIAs detected C. sakazakii in PIF samples artificially contaminated with 1 to 10 bacterial cells per 10 g of sample after 15 h of preenrichment. The use of these serotype-specific MAbs not only allows the reliable detection of C. sakazakii strains but also enables simultaneous serotyping in a simple sandwich EIA method. PMID:26850303

  10. Diversity and Antimicrobial Resistance of Salmonella enterica Isolates from Surface Water in Southeastern United States

    PubMed Central

    Vellidis, George; Liu, Huanli; Jay-Russell, Michele; Zhao, Shaohua; Hu, Zonglin; Wright, Anita; Elkins, Christopher A.

    2014-01-01

    A study of prevalence, diversity, and antimicrobial resistance of Salmonella enterica in surface water in the southeastern United States was conducted. A new scheme was developed for recovery of Salmonella from irrigation pond water and compared with the FDA's Bacteriological Analytical Manual (8th ed., 2014) (BAM) method. Fifty-one isolates were recovered from 10 irrigation ponds in produce farms over a 2-year period; nine Salmonella serovars were identified by pulsed-field gel electrophoresis analysis, and the major serovar was Salmonella enterica serovar Newport (S. Newport, n = 29), followed by S. enterica serovar Enteritidis (n = 6), S. enterica serovar Muenchen (n = 4), S. enterica serovar Javiana (n = 3), S. enterica serovar Thompson (n = 2), and other serovars. It is noteworthy that the PulseNet patterns of some of the isolates were identical to those of the strains that were associated with the S. Thompson outbreaks in 2010, 2012, and 2013, S. Enteritidis outbreaks in 2011 and 2013, and an S. Javiana outbreak in 2012. Antimicrobial susceptibility testing confirmed 16 S. Newport isolates of the multidrug resistant-AmpC (MDR-AmpC) phenotype, which exhibited resistance to ampicillin, chloramphenicol, streptomycin, sulfamethoxazole, and tetracycline (ACSSuT), and to the 1st, 2nd, and 3rd generations of cephalosporins (cephalothin, amoxicillin-clavulanic acid, and ceftriaxone). Moreover, the S. Newport MDR-AmpC isolates had a PFGE pattern indistinguishable from the patterns of the isolates from clinical settings. These findings suggest that the irrigation water may be a potential source of contamination of Salmonella in fresh produce. The new Salmonella isolation scheme significantly increased recovery efficiency from 21.2 (36/170) to 29.4% (50/170) (P = 0.0002) and streamlined the turnaround time from 5 to 9 days with the BAM method to 4 days and thus may facilitate microbiological analysis of environmental water. PMID:25107969

  11. Resident bacteria on leaves enhance survival of immigrant cells of Salmonella enterica.

    PubMed

    Poza-Carrion, Cesar; Suslow, Trevor; Lindow, Steven

    2013-04-01

    Although Salmonella enterica apparently has comparatively low epiphytic fitness on plants, external factors that would influence its ability to survive on plants after contamination would be of significance in the epidemiology of human diseases caused by this human pathogen. Viable population sizes of S. enterica applied to plants preinoculated with Pseudomonas syringae or either of two Erwinia herbicola strains was ≥10-fold higher than that on control plants that were not precolonized by such indigenous bacteria when assessed 24 to 72 h after the imposition of desiccation stress. The protective effect of P. fluorescens, which exhibited antibiosis toward S. enterica in vitro, was only ≈50% that conferred by other bacterial strains. Although S. enterica could produce small cellular aggregates after incubation on wet leaves for several days, and the cells in such aggregates were less susceptible to death upon acute dehydration than solitary cells (as determined by propidium iodide staining), most Salmonella cells were found as isolated cells when it was applied to leaves previously colonized by other bacterial species. The proportion of solitary cells of S. enterica coincident with aggregates of cells of preexisting epiphytic species that subsequently were judged as nonviable by viability staining on dry leaves was as much as 10-fold less than those that had landed on uncolonized portions of the leaf. Thus, survival of immigrant cells of S. enterica on plants appears to be strongly context dependent, and the presence of common epiphytic bacteria on plants can protect such immigrants from at least one key stress (i.e., desiccation) encountered on leaf surfaces. PMID:23506362

  12. Core Oligosaccharide of Plesiomonas shigelloides PCM 2231 (Serotype O17) Lipopolysaccharide—Structural and Serological Analysis

    PubMed Central

    Maciejewska, Anna; Lukasiewicz, Jolanta; Kaszowska, Marta; Man-Kupisinska, Aleksandra; Jachymek, Wojciech; Lugowski, Czeslaw

    2013-01-01

    The herein presented complete structure of the core oligosaccharide of lipopolysaccharide (LPS) P. shigelloides Polish Collection of Microorganisms (PCM) 2231 (serotype O17) was investigated by 1H, 13C NMR spectroscopy, mass spectrometry, chemical analyses and serological methods. The core oligosaccharide is composed of an undecasaccharide, which represents the second core type identified for P. shigelloides serotype O17 LPS. This structure is similar to that of the core oligosaccharide of P. shigelloides strains 302-73 (serotype O1) and 7-63 (serotype O17) and differs from these only by one sugar residue. Serological screening of 55 strains of P. shigelloides with the use of serum against identified core oligosaccharide conjugated with bovine serum albumin (BSA) indicated the presence of similar structures in the LPS core region of 28 O-serotypes. This observation suggests that the core oligosaccharide structure present in strain PCM 2231 could be the most common type among P. shigelloides lipopolysaccharides. PMID:23389090

  13. Molecular characterization and analysis of high-level multidrug-resistance of Shigella flexneri serotype 4s strains from China

    PubMed Central

    Yang, Chaojie; Li, Peng; Zhang, Xiujuan; Ma, Qiuxia; Cui, Xianyan; Li, Hao; Liu, Hongbo; Wang, Jian; Xie, Jing; Wu, Fuli; Sheng, Chunyu; Du, Xinying; Qi, Lihua; Su, Wenli; Jia, Leili; Xu, Xuebin; Zhao, Jiayong; Xia, Shengli; Zhou, Na; Ma, Hui; Qiu, Shaofu; Song, Hongbin

    2016-01-01

    To conduct the first comprehensive analysis of Shigella flexneri serotype 4s, a novel serotype found in 2010, we identified 24 serotype 4s isolates from 1973 shigellosis cases in China (2002–2014). The isolates were characterized by single nucleotide polymorphism (SNP) phylogenetic analysis, pulsed-field gel electrophoresis (PFGE) and multilocus sequence typing (MLST) to determine their genetic relatedness, and analysed further for their antimicrobial susceptibilities and antimicrobial resistance determinants. The PFGE and SNP phylogenetic analyses suggest that S. flexneri serotype 4s strains are derived from multiple serotypes, including two predominant serotypes in China: serotype X variant and serotype II. Three new sequence types were identified by MLST. All isolates were resistant to ticarcillin, ampicillin and tetracycline, with high-level resistance to third-generation cephalosporins. Notably, all the isolates were multidrug resistant (MDR), with the highest levels of resistance observed for eight antimicrobials classes. Most isolates contain various antimicrobial resistance determinants. In conclusion, we found that serotype 4s isolates have multiple evolutionary sources, diverse biochemical characteristics and genomes, and highly prevalent multidrug resistance and antimicrobial-resistant determinants. With few clinical treatment options, continuous monitoring and timely intervention against this emerging MDR serotype is essential. The possibility that serotype 4s will become the next predominant serotype exists. PMID:27374009

  14. Molecular characterization and analysis of high-level multidrug-resistance of Shigella flexneri serotype 4s strains from China.

    PubMed

    Yang, Chaojie; Li, Peng; Zhang, Xiujuan; Ma, Qiuxia; Cui, Xianyan; Li, Hao; Liu, Hongbo; Wang, Jian; Xie, Jing; Wu, Fuli; Sheng, Chunyu; Du, Xinying; Qi, Lihua; Su, Wenli; Jia, Leili; Xu, Xuebin; Zhao, Jiayong; Xia, Shengli; Zhou, Na; Ma, Hui; Qiu, Shaofu; Song, Hongbin

    2016-01-01

    To conduct the first comprehensive analysis of Shigella flexneri serotype 4s, a novel serotype found in 2010, we identified 24 serotype 4s isolates from 1973 shigellosis cases in China (2002-2014). The isolates were characterized by single nucleotide polymorphism (SNP) phylogenetic analysis, pulsed-field gel electrophoresis (PFGE) and multilocus sequence typing (MLST) to determine their genetic relatedness, and analysed further for their antimicrobial susceptibilities and antimicrobial resistance determinants. The PFGE and SNP phylogenetic analyses suggest that S. flexneri serotype 4s strains are derived from multiple serotypes, including two predominant serotypes in China: serotype X variant and serotype II. Three new sequence types were identified by MLST. All isolates were resistant to ticarcillin, ampicillin and tetracycline, with high-level resistance to third-generation cephalosporins. Notably, all the isolates were multidrug resistant (MDR), with the highest levels of resistance observed for eight antimicrobials classes. Most isolates contain various antimicrobial resistance determinants. In conclusion, we found that serotype 4s isolates have multiple evolutionary sources, diverse biochemical characteristics and genomes, and highly prevalent multidrug resistance and antimicrobial-resistant determinants. With few clinical treatment options, continuous monitoring and timely intervention against this emerging MDR serotype is essential. The possibility that serotype 4s will become the next predominant serotype exists. PMID:27374009

  15. Variable abattoir conditions affect Salmonella enterica prevalence and meat quality in swine and pork.

    PubMed

    Hurd, H S; Gailey, J K; McKean, J D; Griffith, R W

    2005-01-01

    Research suggests that abattoir holding pens pose significant Salmonella enterica risk to swine immediately preharvest. The goal of this study was to evaluate those factors related to holding that increased the prevalence of S. enterica in swine at slaughter. To accomplish this goal, we focused on holding time and flooring. Our objectives were to (1) compare Salmonella enterica prevalence among pigs held for short (15-45 min) versus long (up to 4 h) periods before slaughter; and (2) determine the impact of flooring (slatted vs. concrete) as it relates to the prevalence of S. enterica. The study consisted of seven repetitions at a large volume (11,000 head/day) Midwest abattoir. Each repetition consisted of one truck load of pigs (n = 170) sorted into one of three groups: (1) animals held for a short time (15-45 min) on solid floors (short-hold); (2) animals held for 4 +/- 0.5 h on slatted floors; and (3) animals held for 4 +/- 0.5 h on solid concrete floors. At slaughter, samples were collected from 30 pigs in each group. Cecal contents (20 mL), feces (20 g), and the ileocecal lymph node were cultured for S. enterica. Additionally, the effect of holding time on meat quality parameters (loin pH at 35 min and 6 h, color, drip loss) was evaluated for the first four replicates. The proportion of S. enterica-positive samples was highest (p < 0.05) in the cecum of pigs held on solid concrete floors (72.4%), and slightly less for pigs held on slatted floors (63.3%). Animals held for less than 45 min before slaughter demonstrated the lowest proportion of S. enterica-positive samples (52.9%). The pig prevalence, as measured by any one of the three samples being positive, was significantly different (p < 0.05) between animals held on solid floors (81%) and those animals held for 45 min or less before slaughter (69%). Meat quality, as measured by multiple parameters, was adversely affected by lack of a rest period. The mean 24-h pH was significantly lower for the short

  16. Complete genome sequence of Salmonella enterica serovar typhimurium bacteriophage SPN1S.

    PubMed

    Shin, Hakdong; Lee, Ju-Hoon; Lim, Jeong-A; Kim, Hyeryen; Ryu, Sangryeol

    2012-01-01

    To understand the interaction between the host of pathogenic Salmonella enterica serovar Typhimurium and its bacteriophage, we isolated the bacteriophage SPN1S. It is a lysogenic phage in the Podoviridae family and uses the O-antigen of lipopolysaccharides (LPS) as a host receptor. Comparative genomic analysis of phage SPN1S and the S. enterica serovar Anatum-specific phage ε15 revealed different host specificities, probably due to the low homology of host specificity-related genes. Here we report the complete circular genome sequence of S. Typhimurium-specific bacteriophage SPN1S and show the results of our analysis. PMID:22205721

  17. Serotype distribution of Streptococcus pneumoniae in children with invasive diseases in Turkey: 2008-2014.

    PubMed

    Ceyhan, Mehmet; Ozsurekci, Yasemin; Gürler, Nezahat; Öksüz, Lütfiye; Aydemir, Sohret; Ozkan, Sengul; Yuksekkaya, Serife; Keser Emiroglu, Melike; Gültekin, Meral; Yaman, Akgün; Kiremitci, Abdurrahman; Yanık, Keramettin; Karli, Arzu; Ozcinar, Hatice; Aydin, Faruk; Bayramoglu, Gulcin; Zer, Yasemin; Gulay, Zeynep; Gayyurhan, Efgan Dogan; Gül, Mustafa; Özakın, Cüneyt; Güdücüoğlu, Hüseyin; Perçin, Duygu; Akpolat, Nezahat; Ozturk, Candan; Camcıoğlu, Yıldız; Karadağ Öncel, Eda; Çelik, Melda; Şanal, Laser; Uslu, Hakan

    2016-01-01

    Successful vaccination policies for protection from invasive pneumococcal diseases (IPD) dependent on determination of the exact serotype distribution in each country. We aimed to identify serotypes of pneumococcal strains causing IPD in children in Turkey and emphasize the change in the serotypes before and after vaccination with 7-valent pneumococcal conjugate vaccine (PCV-7) was included and PCV-13 was newly changed in Turkish National Immunization Program. Streptococcus pneumoniae strains were isolated at 22 different hospitals of Turkey, which provide healthcare services to approximately 65% of the Turkish population. Of the 335 diagnosed cases with S. pneumoniae over the whole period of 2008-2014, the most common vaccine serotypes were 19F (15.8%), 6B (5.9%), 14 (5.9%), and 3 (5.9%). During the first 5 y of age, which is the target population for vaccination, the potential serotype coverage ranged from 57.5 % to 36.8%, from 65.0% to 44.7%, and from 77.4% to 60.5% for PCV-7, PCV-10, and PCV-13 in 2008-2014, respectively. The ratio of non-vaccine serotypes was 27.2% in 2008-2010 whereas was 37.6% in 2011-2014 (p=0.045). S. penumoniae serotypes was less non-susceptible to penicillin as compared to our previous results (33.7 vs 16.5 %, p=0.001). The reduction of those serotype coverage in years may be attributed to increasing vaccinated children in Turkey and the increasing non-vaccine serotype may be explained by serotype replacement. Our ongoing IPD surveillance is a significant source of information for the decision-making processes on pneumococcal vaccination. PMID:26325175

  18. Non-invasive pneumococcal pneumonia in Portugal--serotype distribution and antimicrobial resistance.

    PubMed

    Horácio, Andreia N; Lopes, Joana P; Ramirez, Mário; Melo-Cristino, José

    2014-01-01

    There is limited information on the serotypes causing non-invasive pneumococcal pneumonia (NIPP). Our aim was to characterize pneumococci causing NIPP in adults to determine recent changes in serotype prevalence, the potential coverage of pneumococcal vaccines and changes in antimicrobial resistance. Serotypes and antimicrobial susceptibility profiles of a sample of 1300 isolates recovered from adult patients (≥18 yrs) between 1999 and 2011 (13 years) were determined. Serotype 3 was the most frequent cause of NIPP accounting for 18% of the isolates. The other most common serotypes were 11A (7%), 19F (7%), 19A (5%), 14 (4%), 22F (4%), 23F (4%) and 9N (4%). Between 1999 and 2011, there were significant changes in the proportion of isolates expressing vaccine serotypes, with a steady decline of the serotypes included in the 7-valent conjugate vaccine from 31% (1999-2003) to 11% (2011) (P<0.001). Taking together the most recent study years (2009-2011), the potential coverage of the 13-valent conjugate vaccine was 44% and of the 23-valent polysaccharide vaccine was 66%. While erythromycin resistance increased from 8% in 1999-2003 to 18% in 2011 (P<0.001), no significant trend was identified for penicillin non-susceptibility, which had an average value of 18.5%. The serotype distribution found in this study for NIPP was very different from the one previously described for IPD, with only two serotypes in common to the ones responsible for half of each presentation in 2009-2011 - serotypes 3 and 19A. In spite of these differences, the overall prevalence of resistant isolates was similar in NIPP and in IPD. PMID:25075961

  19. Risk factors for Salmonella enterica subsp. enterica shedding by market-age pigs in French farrow-to-finish herds.

    PubMed

    Beloeil, P-A; Fravalo, P; Fablet, C; Jolly, J-P; Eveno, E; Hascoet, Y; Chauvin, C; Salvat, G; Madec, F

    2004-04-30

    Fattening-pigs carriers of Salmonella enterica are believed to be a main source of carcass and pork contamination at the later steps of the meat process. We did a prospective study in 2000-2001 in 105 French farrow-to-finish pig farms. In each farm, a batch of contemporary fattening pigs housed in the same room was followed throughout the fattening period. Salmonella shedding was assessed on environmental samples of faecal material (taken by means of pairs of gauze socks) analysed by classical bacteriological methods. 36.2% of the batches studied had at least one contaminated environmental sample and therefore were classified as Salmonella-shedding batches. Logistic regression was used to assess the association between managerial and hygiene practices and health status and the shedding risk at the end of the finishing period. Emptying the pit below the slatted floor after the previous batch of sows was removed and frequent removal of sow dung during the lactation period were protective. Presence of residual Salmonella contamination of the floor and pen partitions in the fattening rooms before loading the growing pigs also was a risk factor. The risk for Salmonella shedding at the end of the fattening period was increased when dry feed (versus wet feed) was provided during the fattening period. Lastly, Lawsonia intracellularis seroconversion and PRRSV seropositivity during the fattening period also was a risk factor for Salmonella shedding. PMID:15099720

  20. Vertical transmission of Salmonella paratyphi A.

    PubMed

    Raveendran, R; Wattal, C; Sharma, A; Kler, N; Garg, P; Gujral, K; Khera, N

    2007-08-01

    Neonatal enteric fever is a rare but life-threatening illness. Patients may present with varying severity, Salmonella enterica serotype Typhi causing more severe illness than Salmonella enterica serotype Paratyphi A. Salmonella enterica serotype Paratyphi A is considered to cause milder infection with fewer complications. We report a rare case of vertical transmission of Salmonella enterica serotype Paratyphi A with severe complications and high mortality. Even though there are case reports of vertical transmission of Salmonella enterica serotype Typhi, to our knowledge, this is the first case report of vertical transmission of Salmonella enterica serotype ParatyphiA. The role of blood culture in accurate diagnosis and treatment is also discussed. PMID:17785907

  1. VP7 from African horse sickness virus serotype 9 protects mice against a lethal, heterologous serotype challenge.

    PubMed

    Wade-Evans, A M; Pullen, L; Hamblin, C; O'Hara, R S; Burroughs, J N; Mertens, P P

    1998-01-01

    An established mouse model system was used to evaluate the effectiveness of the major outer core protein VP7 of African horse sickness virus (AHSV) serotype 9 as a subunit vaccine. Balb C mice were immunised with VP7 crystals purified from AHSV infected BHK cells. In groups of mice, each of which was immunised with > or = 1.5 micrograms of the protein in Freund's adjuvant, > or = 80% of mice survived challenge with a virulent strain of a heterologous AHSV serotype (AHSV 7), that killed > or = 80% of the mice in the uninoculated control groups. This level of protection was significantly greater than that observed in mice inoculated with equivalent amounts of either denatured VP7 (50% survival), or GST/VP7 fusion protein (50-70% survival), or which were vaccinated with AHSV 9 (40-50% survival). The VP7 protein folding, or its assembly into crystals, are thought to play some role in the effectiveness of the protective response observed. Titres of circulating antibodies against AHSV VP7 were determined by competitive ELISA but did not appear to correlate with the levels of protection observed. Passive transfer of these antibodies to syngeneic recipients also failed to protect Balb C mice from the AHSV 7 challenge. The observed protection is therefore unlikely to be due to an antibody mediated immune response. PMID:9785508

  2. Evaluation of Salmonella serotype distributions from commercial broiler hatcheries and grower houses.

    PubMed

    Byrd, J A; DeLoach, J R; Corrier, D E; Nisbet, D J; Stanker, L H

    1999-01-01

    By conventional trayliner (hatcheries) and drag swab assembly (broiler houses) culture methods, the isolation distribution of Salmonella serotypes from five commercial broiler hatcheries (three sample times) and 13 broiler farms (eight sample times) was evaluated. A total of 11 different Salmonella serotypes were isolated from hatcheries, with Salmonella heidelberg (9/30) and Salmonella kentucky (6/30) accounting for 50% of the total isolations. Of 700 chick paperpad trayliners sampled, regardless of lot (breeder flock source) or hatchery, 12% were positive for Salmonella. When 10 individual trayliners were cultured from individual lots (same breeder flock source), Salmonella was detected in 24/57 lots (42%). Multiple serotypes were simultaneously isolated from the same lot on three occasions (6%). Of the 21 lots that were serially sampled, the Salmonella serotype detected was different within lots eight times (38%) on at least one occasion of two or more sampling times. Of the 196 individual broiler houses sampled, 44 were positive for Salmonella (42%). Twelve different serotypes were isolated from broiler houses during this study. The serotypes isolated most frequently were S. heidelberg (34/94) and S. kentucky (22/94). These two serotypes accounted for 59.6% (56/94) of the total broiler house isolations. Of the 38 houses that were serially sampled, two or more serotypes were detected in the same broiler house on 20 occasions (53%). Of the 38 serially sampled houses (four or more times), a consistent Salmonella serotype was detected in five houses (13%). In only 5 of the 38 (13%) serially sampled houses did we fail to detect Salmonella on four or more samplings. No significant difference in Salmonella isolation frequency was observed between poultry houses using new or used litter. These data support previous findings indicating that paratyphoid Salmonella serotypes are prevalent in some broiler hatcheries and houses. Further, the observation of multiple

  3. Determination of Pneumococcal Serotypes in Meningitis Cases in Niger, 2003–2011

    PubMed Central

    Collard, Jean-Marc; Alio Sanda, Abdel-kader; Jusot, Jean-François

    2013-01-01

    Background The epidemiology of pneumococcal meningitis in the African ‘meningitis belt’ is poorly studied. In order to ensure an effective vaccination strategy and post-vaccination surveillance, we examined the serotype distribution patterns of pneumococcal meningitis in Niger over the period 2003–2011. Methods Cerebrospinal fluid (CSF) samples were collected from different health facilities throughout Niger in the frame of the national microbiological surveillance of meningitis. Determination of the serotype of CSF positive for pneumococci was performed using a sequential multiplex PCR method (SM-PCR) adapted with a national algorithm in which 32 different serotypes were covered and grouped into eight consecutive PCR. Results The SM-PCR assay could predict the Sp serotype for 779 CSF (88.7%), 98 CSF (11.3%) were not-typeable in our national-adapted algorithm. In total, 26 different serotypes were identified. Serotype 1 (n = 393) was the most prevalent and accounted for 45.3% of infections, followed by serogroups/serotypes 12F/(12A)/(44)/(46) (7.3%), 6/(6A/6B/6C/6D) (5.4%), 14 (5.2%), 5 (4.6%), 23F (4.2%), 45 (3.6%), 2 (3.1%), 18/(18A/18B/18C/18F) (2.9%) and 17 others serotypes with a prevalence of less than 2%. The proportion of serotype 1 in infants(<2 years old) represented only 4.3% of the cases affected by this serotype. In contrast, serotypes 5, 6, 14, 19A and 23F were only detected in very young children. Conclusions The proportion of serotype 1 in the pneumococcal meningitis cases and the theoretical vaccine coverage across all age groups advocates for the introduction of a conjugate vaccine (PCV10 or 13) into the Expanded Programme on Immunization (EPI) in Niger. Post-vaccine introduction surveillance supported by molecular approaches will be essential to provide a comprehensive picture of the impact of the vaccine on the burden reduction of pneumococcal meningitis and on pneumococcal serotype distribution. PMID:23555971

  4. A novel high-throughput method for molecular serotyping and serotype-specific quantification of Streptococcus pneumoniae using a nanofluidic real-time PCR system.

    PubMed

    Dhoubhadel, Bhim Gopal; Yasunami, Michio; Yoshida, Lay-Myint; Thi, Hien Anh Nguyen; Thi, Thu Huong Vu; Thi, Thuy Ai Nguyen; Watanabe, Kiwao; Suzuki, Motoi; Morimoto, Konosuke; Dang, Duc Anh; Ariyoshi, Koya

    2014-04-01

    Serotype-specific quantification data are essential for elucidating the complex epidemiology of Streptococcus pneumoniae and evaluating pneumococcal vaccine efficacy. Various PCR-based assays have been developed to circumvent the drawback of labour-intensive and time-consuming culture-based procedures for serotype determination and quantification of pneumococcus. Here, we applied a nanofluidic real-time PCR system to establish a novel assay. Twenty-nine primer pairs, 13 of which were newly designed, were selected for the assay to cover 50 serotypes including all currently available conjugate and polysaccharide vaccine serotypes. All primer pairs were evaluated for their sensitivity, specificity, efficiency, repeatability, accuracy and reproducibility on the Fluidigm Biomark HD System, a nanofluidic real-time PCR system, by drawing standard curves with a serial dilution of purified DNA. We applied the assay to 52 nasopharyngeal swab samples from patients with pneumonia confirmed by chest X-ray to validate its accuracy. Minimum detection levels of this novel assay using the nanofluidic real-time PCR system were comparable to the conventional PCR-based assays (between 30 and 300 copies per reaction). They were specific to their targets with good repeatability (sd of copy number of 0.1), accuracy (within ±0.1 fold difference in log10 copy number) and reproducibility (sd of copy number of 0.1). When artificially mixed DNA samples consisting of multiple serotypes in various ratios were tested, all the serotypes were detected proportionally, including a minor serotype of one in 1000 copies. In the nasopharyngeal samples, the PCR system detected all the culture-positive samples and 22 out of 23 serotypes identified by the conventional method were matched with PCR results. We conclude that this novel assay, which is able to differentially quantify 29 pneumococcus groups for 45 test samples in a single run, is applicable to the large-scale epidemiological study of

  5. The association of serotype and pulsed-field gel electrophoresis genotype in isolates of Streptococcus pneumoniae isolated in Israel.

    PubMed

    Bar-Meir, M; Naaman, G; Assous, M; Korenman, Z; Valinsky, L; Picard, E

    2015-05-01

    The relationship between Streptococcus pneumoniae isolates causing invasive infections in children admitted to a single center in central Israel was examined by pulsed-field gel electrophoresis (PFGE) and serotyping. Although there was a close correlation between serotype and PFGE clone, the genetic diversity varied by serotype, with some genotypes comprising multiple serotypes. Additionally, clones C and D were associated with higher penicillin minimum inhibitory concentrations. Serotyping alone may be insufficient for epidemiological mapping of pneumococcal isolates in the era of pneumococcal conjugate polysaccharide vaccines. PMID:25749648

  6. IncI1 Plasmids Carrying Various blaCTX-M Genes Contribute to Ceftriaxone Resistance in Salmonella enterica Serovar Enteritidis in China

    PubMed Central

    Kan, Biao; Chan, Edward Wai-chi

    2015-01-01

    Resistance to extended-spectrum β-lactams in Salmonella, in particular, in serotypes such as Salmonella enterica serovar Enteritidis that are frequently associated with clinical infections, is a serious public health concern. In this study, phenotypic characterization of 433 clinical S. Enteritidis strains obtained from a nationwide collection of the Chinese Center for Disease Control and Prevention during the period from 2005 to 2010 depicted a trend of increasing resistance to ceftriaxone from 2008 onwards. Seventeen (4%) of the strains were found to be resistant to ceftriaxone, 7% were found to be resistant to ciprofloxacin, and 0.7% were found to be resistant to both ciprofloxacin and ceftriaxone. Most of the ceftriaxone-resistant S. Enteritidis strains (15/17) were genetically unrelated and originated from Henan Province. The complete sequence of an IncI1 plasmid, pSE115, which belonged to a novel sequence type, was obtained. This 87,255-bp IncI1 plasmid was found to harbor a blaCTX-M-14 gene in a novel multidrug resistance region (MRR) within the tra locus. Although the majority of strains were also found to contain conjugative IncI1 plasmids with a size similar to that of pSE115 (∼90 kb) and harbor a variety of blaCTX-M group 1 and group 9 elements, the novel MRR site at the tra locus in pSE115 was not detectable in the other IncI1 plasmids. The findings from this study show that cephalosporin resistance in S. Enteritidis strains collected in China was mainly due to the dissemination of IncI1 plasmids carrying blaCTX-M, resembling the situation in which IncI1 plasmids serve as major vectors of blaCTX-M variants in other members of the Enterobacteriaceae. PMID:26643327

  7. Emergence and Prevalence of Non-H2S-Producing Salmonella enterica Serovar Senftenberg Isolates Belonging to Novel Sequence Type 1751 in China

    PubMed Central

    Yi, Shengjie; Xie, Jing; Liu, Nan; Li, Peng; Xu, Xuebin; Li, Hao; Sun, Jichao; Wang, Jian; Liang, Beibei; Yang, Chaojie; Wang, Xu; Hao, Rongzhang; Wang, Ligui; Wu, Zhihao; Zhang, Jianmin; Wang, Yong; Huang, Liuyu; Sun, Yansong; Klena, John D.; Meng, Jianghong

    2014-01-01

    Salmonella enterica serovar Senftenberg is a common nontyphoidal Salmonella serotype which causes human Salmonella infections worldwide. In this study, 182 S. Senftenberg isolates, including 17 atypical non-hydrogen sulfide (H2S)-producing isolates, were detected in China from 2005 to 2011. The microbiological and genetic characteristics of the non-H2S-producing and selected H2S-producing isolates were determined by using pulsed-field gel electrophoresis (PFGE), multilocus sequence typing (MLST), and clustered regularly interspaced short palindromic repeat (CRISPR) analysis. The phs operons were amplified and sequenced. The 17 non-H2S-producing and 36 H2S-producing isolates belonged to 7 sequence types (STs), including 3 new STs, ST1751, ST1757, and ST1758. Fourteen of the 17 non-H2S-producing isolates belonged to ST1751 and had very similar PFGE patterns. All 17 non-H2S-producing isolates had a nonsense mutation at position 1621 of phsA. H2S-producing and non-H2S-producing S. Senftenberg isolates were isolated from the same stool sample from three patients; isolates from the same patients displayed the same antimicrobial susceptibility, ST, and PFGE pattern but could be discriminated based on CRISPR spacers. Non-H2S-producing S. Senftenberg isolates belonging to ST1751 have been prevalent in Shanghai, China. It is possible that these emerging organisms will disseminate further, because they are difficult to detect. Thus, we should strengthen the surveillance for the spread of this atypical S. Senftenberg variant. PMID:24829240

  8. Virulence Characterization of Salmonella enterica by a New Microarray: Detection and Evaluation of the Cytolethal Distending Toxin Gene Activity in the Unusual Host S. Typhimurium

    PubMed Central

    Figueiredo, Rui; Card, Roderick; Nunes, Carla; AbuOun, Manal; Bagnall, Mary C.; Nunez, Javier; Mendonça, Nuno; Anjum, Muna F.; da Silva, Gabriela Jorge

    2015-01-01

    Salmonella enterica is a zoonotic foodborne pathogen that causes acute gastroenteritis in humans. We assessed the virulence potential of one-hundred and six Salmonella strains isolated from food animals and products. A high through-put virulence genes microarray demonstrated Salmonella Pathogenicity Islands (SPI) and adherence genes were highly conserved, while prophages and virulence plasmid genes were variably present. Isolates were grouped by serotype, and virulence plasmids separated S. Typhimurium in two clusters. Atypical microarray results lead to whole genome sequencing (WGS) of S. Infantis Sal147, which identified deletion of thirty-eight SPI-1 genes. Sal147 was unable to invade HeLa cells and showed reduced mortality in Galleria mellonella infection model, in comparison to a SPI-1 harbouring S. Infantis. Microarray and WGS of S. Typhimurium Sal199, established for the first time in S. Typhimurium presence of cdtB and other Typhi-related genes. Characterization of Sal199 showed cdtB genes were upstream of transposase IS911, and co-expressed with other Typhi-related genes. Cell cycle arrest, cytoplasmic distension, and nuclear enlargement were detected in HeLa cells infected by Sal199, but not with S. Typhimurium LT2. Increased mortality of Galleria was detected on infection with Sal199 compared to LT2. Thus, Salmonella isolates were rapidly characterized using a high through-put microarray; helping to identify unusual virulence features which were corroborated by further characterisation. This work demonstrates that the use of suitable screening methods for Salmonella virulence can help assess the potential risk associated with certain Salmonella to humans. Incorporation of such methodology into surveillance could help reduce the risk of emergence of epidemic Salmonella strains. PMID:26244504

  9. Extensive genetic variability linked to IS26 insertions in the fljB promoter region of atypical monophasic variants of Salmonella enterica serovar Typhimurium.

    PubMed

    Boland, Cécile; Bertrand, Sophie; Mattheus, Wesley; Dierick, Katelijne; Jasson, Vicky; Rosseel, Toon; Van Borm, Steven; Mahillon, Jacques; Wattiau, Pierre

    2015-05-01

    Fifty-nine monophasic Salmonella enterica serovar Typhimurium isolates, collected in Belgium during the period from 2008 to 2011, have been serotyped as 4,[5]:i:- and shown to harbor an fljB coding sequence. The genetic differences between these strains and phenotypically biphasic Salmonella Typhimurium were analyzed through PCR and DNA sequencing. Genetic alterations in the fljB promoter region affecting expression of the phase 2 flagellin were observed in 53 isolates. Other genetic events in the invertible region carrying the fljB promoter were observed in 2 isolates. For the remaining 4 isolates, no molecular differences with a reference biphasic Salmonella Typhimurium strain could be observed. Next-generation sequencing of one representative isolate affected in the fljB promoter region revealed a 26-kb IS26 composite transposon insertion along with a local genomic rearrangement. Several other IS26 element-mediated alterations of this genomic region were observed. This group of monophasic Salmonella Typhimurium isolates was genetically heterogeneous, as revealed by multilocus variable-number tandem-repeat analysis (MLVA), PCR, and sequencing. Pigs and pork represented a major source of such monophasic isolates in Belgium, as reported in other countries. Three out of 5 isolates of human origin presented genetic profiles identical to those of food isolates, demonstrating the pathogenic potential of the newly characterized variants and potential dissemination along the food chain. This study highlighted the key role played by IS26 insertions in the loss of phase 2 flagellin expression and the subsequent generation of multiple monophasic variant lineages from biphasic Salmonella Typhimurium ancestors. PMID:25724958

  10. Variable Carbon Catabolism among Salmonella enterica Serovar Typhi Isolates

    PubMed Central

    Chai, Lay Ching; Kong, Boon Hong; Elemfareji, Omar Ismail; Thong, Kwai Lin

    2012-01-01

    Background Salmonella enterica serovar Typhi (S. Typhi) is strictly a human intracellular pathogen. It causes acute systemic (typhoid fever) and chronic infections that result in long-term asymptomatic human carriage. S. Typhi displays diverse disease manifestations in human infection and exhibits high clonality. The principal factors underlying the unique lifestyle of S. Typhi in its human host during acute and chronic infections remain largely unknown and are therefore the main objective of this study. Methodology/Principal Findings To obtain insight into the intracellular lifestyle of S. Typhi, a high-throughput phenotypic microarray was employed to characterise the catabolic capacity of 190 carbon sources in S. Typhi strains. The success of this study lies in the carefully selected library of S. Typhi strains, including strains from two geographically distinct areas oftyphoid endemicity, an asymptomatic human carrier, clinical stools and blood samples and sewage-contaminated rivers. An extremely low carbon catabolic capacity (27% of 190 carbon substrates) was observed among the strains. The carbon catabolic profiles appeared to suggest that S. Typhi strains survived well on carbon subtrates that are found abundantly in the human body but not in others. The strains could not utilise plant-associated carbon substrates. In addition, α-glycerolphosphate, glycerol, L-serine, pyruvate and lactate served as better carbon sources to monosaccharides in the S. Typhi strains tested. Conclusion The carbon catabolic profiles suggest that S. Typhi could survive and persist well in the nutrient depleted metabolic niches in the human host but not in the environment outside of the host. These findings serve as caveats for future studies to understand how carbon catabolism relates to the pathogenesis and transmission of this pathogen. PMID:22662115

  11. Global Genomic Epidemiology of Salmonella enterica Serovar Typhimurium DT104

    DOE PAGESBeta

    Leekitcharoenphon, Pimlapas; Hendriksen, Rene S.; Le Hello, Simon; Weill, François-Xavier; Baggesen, Dorte Lau; Jun, Se-Ran; Ussery, David W.; Lund, Ole; Crook, Derrick W.; Wilson, Daniel J.; et al

    2016-03-04

    It has been 30 years since the initial emergence and subsequent rapid global spread of multidrug-resistant Salmonella enterica serovar Typhimurium DT104 (MDR DT104). Nonetheless, its origin and transmission route have never been revealed. In this paper, we used whole-genome sequencing (WGS) and temporally structured sequence analysis within a Bayesian framework to reconstruct temporal and spatial phylogenetic trees and estimate the rates of mutation and divergence times of 315 S. Typhimurium DT104 isolates sampled from 1969 to 2012 from 21 countries on six continents. DT104 was estimated to have emerged initially as antimicrobial susceptible in ~1948 (95% credible interval [CI], 1934more » to 1962) and later became MDR DT104 in ~1972 (95% CI, 1972 to 1988) through horizontal transfer of the 13-kb Salmonella genomic island 1 (SGI1) MDR region into susceptible strains already containing SGI1. This was followed by multiple transmission events, initially from central Europe and later between several European countries. An independent transmission to the United States and another to Japan occurred, and from there MDR DT104 was probably transmitted to Taiwan and Canada. An independent acquisition of resistance genes took place in Thailand in ~1975 (95% CI, 1975 to 1990). In Denmark, WGS analysis provided evidence for transmission of the organism between herds of animals. Interestingly, the demographic history of Danish MDR DT104 provided evidence for the success of the program to eradicate Salmonella from pig herds in Denmark from 1996 to 2000. Finally, the results from this study refute several hypotheses on the evolution of DT104 and suggest that WGS may be useful in monitoring emerging clones and devising strategies for prevention of Salmonella infections.« less

  12. Stress Response of Salmonella enterica Serovar Typhimurium to Acidified Nitrite

    PubMed Central

    Mühlig, Anna; Behr, Jürgen; Scherer, Siegfried

    2014-01-01

    The antimicrobial action of the curing agent sodium nitrite (NaNO2), which is added as a preservative to raw meat products, depends on its conversion to nitric oxide and other reactive nitrogen species under acidic conditions. In this study, we used RNA sequencing to analyze the acidified-NaNO2 shock and adaptive responses of Salmonella enterica serovar Typhimurium, a frequent contaminant in raw meat, considering parameters relevant for the production of raw-cured sausages. Upon a 10-min exposure to 150 mg/liter NaNO2 in LB (pH 5.5) acidified with lactic acid, genes involved in nitrosative-stress protection, together with several other stress-related genes, were induced. In contrast, genes involved in translation, transcription, replication, and motility were downregulated. The induction of stress tolerance and the reduction of cell proliferation obviously promote survival under harsh acidified-NaNO2 stress. The subsequent adaptive response was characterized by upregulation of NsrR-regulated genes and iron uptake systems and by downregulation of genes involved in anaerobic respiratory pathways. Strikingly, amino acid decarboxylase systems, which contribute to acid tolerance, displayed increased transcript levels in response to acidified NaNO2. The induction of systems known to be involved in acid resistance indicates a nitrite-mediated increase in the level of acid stress. Deletion of cadA, which encodes lysine decarboxylase, resulted in increased sensitivity to acidified NaNO2. Intracellular pH measurements using a pH-sensitive green fluorescent protein (GFP) variant showed that the cytoplasmic pH of S. Typhimurium in LB medium (pH 5.5) is decreased upon the addition of NaNO2. This study provides the first evidence that intracellular acidification is an additional antibacterial mode of action of acidified NaNO2. PMID:25107963

  13. Global Genomic Epidemiology of Salmonella enterica Serovar Typhimurium DT104

    PubMed Central

    Hendriksen, Rene S.; Le Hello, Simon; Weill, François-Xavier; Baggesen, Dorte Lau; Jun, Se-Ran; Lund, Ole; Crook, Derrick W.; Wilson, Daniel J.; Aarestrup, Frank M.

    2016-01-01

    It has been 30 years since the initial emergence and subsequent rapid global spread of multidrug-resistant Salmonella enterica serovar Typhimurium DT104 (MDR DT104). Nonetheless, its origin and transmission route have never been revealed. We used whole-genome sequencing (WGS) and temporally structured sequence analysis within a Bayesian framework to reconstruct temporal and spatial phylogenetic trees and estimate the rates of mutation and divergence times of 315 S. Typhimurium DT104 isolates sampled from 1969 to 2012 from 21 countries on six continents. DT104 was estimated to have emerged initially as antimicrobial susceptible in ∼1948 (95% credible interval [CI], 1934 to 1962) and later became MDR DT104 in ∼1972 (95% CI, 1972 to 1988) through horizontal transfer of the 13-kb Salmonella genomic island 1 (SGI1) MDR region into susceptible strains already containing SGI1. This was followed by multiple transmission events, initially from central Europe and later between several European countries. An independent transmission to the United States and another to Japan occurred, and from there MDR DT104 was probably transmitted to Taiwan and Canada. An independent acquisition of resistance genes took place in Thailand in ∼1975 (95% CI, 1975 to 1990). In Denmark, WGS analysis provided evidence for transmission of the organism between herds of animals. Interestingly, the demographic history of Danish MDR DT104 provided evidence for the success of the program to eradicate Salmonella from pig herds in Denmark from 1996 to 2000. The results from this study refute several hypotheses on the evolution of DT104 and suggest that WGS may be useful in monitoring emerging clones and devising strategies for prevention of Salmonella infections. PMID:26944846

  14. Genetic and virulence-phenotype characterization of serotypes 2 and 9 of Streptococcus suis swine isolates.

    PubMed

    Blume, Verena; Luque, Inmaculada; Vela, Ana I; Borge, Carmen; Maldonado, Alfonso; Domínguez, Lucas; Tarradas, Carmen; Fernández-Garayzábal, José F

    2009-09-01

    The aim of this study was to analyze the genetic characteristics and virulence phenotypes of Streptococcus suis, specifically, in clinical isolates of serotypes 2 and 9 (n = 195), obtained from diverse geographical areas across Spain. Pulsed-field gel electrophoresis (PFGE) typing identified 97 genetic profiles, 68% of which were represented by single isolates, indicative of a substantial genetic diversity among the S. suis isolates analyzed. Five PFGE profiles accounted for 33.3% of the isolates and were isolated from 38% of the herds in nine different provinces, indicative of the bacterium's widespread distribution in the Spanish swine population. Representative isolates of the most prevalent PFGE profiles of both serotypes were subjected to multilocus sequence typing (MLST) analysis. The results indicated that serotypes 2 and 9 have distinct genetic backgrounds. Serotype 2 isolates belong to the ST1 complex, a highly successful clone that has spread over most European countries. In accordance with isolates of this complex, most serotype 2 isolates also expressed the phenotype MRP(+)EF(+)SLY(+). Serotype 9 isolates belong to the ST61 complex, which is distantly related to the widespread European ST87 clone. Also, in contrast to most isolates of the European ST87 clone, which express the large variant MRP*, the majority of serotype 9 isolates (97.9%) did not express the protein. PMID:19784922

  15. Emergence and Distribution of Foot-and-Mouth Disease Virus Serotype A and O in Bangladesh.

    PubMed

    Nandi, S P; Rahman, M Z; Momtaz, S; Sultana, M; Hossain, M A

    2015-06-01

    Foot-and-mouth disease (FMD) is endemic in Bangladesh and is predominantly due to FMDV serotype O. In 2012, FMD outbreaks were identified in five different districts of Bangladesh. Of 56 symptomatic cattle epithelial tissue samples, diagnostic PCR assay based on 5'-URT detected 38 FMDV infections. Viral genotyping targeting VP1-encoding region confirmed emergence of two distinct serotypes, A and O with an abundance of serotype A in Chittagong and Gazipur districts and serotype O in Pabna and Faridpur. Only single lineage of both A and O was retrieved from samples of five different regions. Sequencing and phylogenetic analysis of VP1 sequences revealed that serotype O sequences were closely related to the Ind 2001 sublineage of Middle East-South Asia (ME-SA) topotype that was previously circulating in Bangladesh, and serotype A sequences belonging to the genotype VII that was dominant in India during the last decade. The results suggest that extensive cross-border animal movement from neighbouring countries is the most likely source of FMDV serotypes in Bangladesh. PMID:23734722

  16. Evaluation of commercial antisera for serotyping heat-labile antigens of Campylobacter jejuni and Campylobacter coli.

    PubMed Central

    Nicholson, M A; Patton, C M

    1993-01-01

    Commercial antisera for serotyping 22 heat-labile antigens of Campylobacter jejuni and Campylobacter coli were evaluated by using 66 isolates from human and nonhuman sources. Test results were compared with results of tests using antisera produced at the Centers for Disease Control (CDC), Atlanta, Ga. All strains (three isolates of each of the 22 serotypes) were typeable with the CDC antisera. Of 66 test strains, 39 (59%) were typed as the same serotype with both sets of antisera. Twenty-four strains (36%), including two heat-labile serotype reference strains, were nonreactive with the commercial antisera, and three strains (4.5%) were typed as serotypes different from those obtained with CDC antisera. Five of the 22 commercial antisera correctly serotyped all homologous strains. Our study indicated that two polyvalent antiserum pools, 7 unabsorbed antisera, and 16 absorbed monovalent antisera are weak and need modification to enhance their antibody titers. Further studies are necessary to explain the antigenic change to a different serotype in three strains. PMID:8463402

  17. Serotype 1 and 8 Pneumococci Evade Sensing by Inflammasomes in Human Lung Tissue

    PubMed Central

    Fatykhova, Diana; Rabes, Anne; Machnik, Christoph; Guruprasad, Kunchur; Pache, Florence; Berg, Johanna; Toennies, Mario; Bauer, Torsten T.; Schneider, Paul; Schimek, Maria; Eggeling, Stephan; Mitchell, Timothy J.; Mitchell, Andrea M.; Hilker, Rolf; Hain, Torsten; Suttorp, Norbert; Hippenstiel, Stefan

    2015-01-01

    Streptococcus pneumoniae is a major cause of pneumonia, sepsis and meningitis. The pore-forming toxin pneumolysin is a key virulence factor of S. pneumoniae, which can be sensed by the NLRP3 inflammasome. Among the over 90 serotypes, serotype 1 pneumococci (particularly MLST306) have emerged across the globe as a major cause of invasive disease. The cause for its particularity is, however, incompletely understood. We therefore examined pneumococcal infection in human cells and a human lung organ culture system mimicking infection of the lower respiratory tract. We demonstrate that different pneumococcal serotypes differentially activate inflammasome-dependent IL-1β production in human lung tissue and cells. Whereas serotype 2, 3, 6B, 9N pneumococci expressing fully haemolytic pneumolysins activate NLRP3 inflammasome-dependent responses, serotype 1 and 8 strains expressing non-haemolytic toxins are poor activators of IL-1β production. Accordingly, purified haemolytic pneumolysin but not serotype 1-associated non-haemolytic toxin activates strong IL-1β production in human lungs. Our data suggest that the evasion of inflammasome-dependent innate immune responses by serotype 1 pneumococci might contribute to their ability to cause invasive diseases in humans. PMID:26317436

  18. Serotype 1 and 8 Pneumococci Evade Sensing by Inflammasomes in Human Lung Tissue.

    PubMed

    Fatykhova, Diana; Rabes, Anne; Machnik, Christoph; Guruprasad, Kunchur; Pache, Florence; Berg, Johanna; Toennies, Mario; Bauer, Torsten T; Schneider, Paul; Schimek, Maria; Eggeling, Stephan; Mitchell, Timothy J; Mitchell, Andrea M; Hilker, Rolf; Hain, Torsten; Suttorp, Norbert; Hippenstiel, Stefan; Hocke, Andreas C; Opitz, Bastian

    2015-01-01

    Streptococcus pneumoniae is a major cause of pneumonia, sepsis and meningitis. The pore-forming toxin pneumolysin is a key virulence factor of S. pneumoniae, which can be sensed by the NLRP3 inflammasome. Among the over 90 serotypes, serotype 1 pneumococci (particularly MLST306) have emerged across the globe as a major cause of invasive disease. The cause for its particularity is, however, incompletely understood. We therefore examined pneumococcal infection in human cells and a human lung organ culture system mimicking infection of the lower respiratory tract. We demonstrate that different pneumococcal serotypes differentially activate inflammasome-dependent IL-1β production in human lung tissue and cells. Whereas serotype 2, 3, 6B, 9N pneumococci expressing fully haemolytic pneumolysins activate NLRP3 inflammasome-dependent responses, serotype 1 and 8 strains expressing non-haemolytic toxins are poor activators of IL-1β production. Accordingly, purified haemolytic pneumolysin but not serotype 1-associated non-haemolytic toxin activates strong IL-1β production in human lungs. Our data suggest that the evasion of inflammasome-dependent innate immune responses by serotype 1 pneumococci might contribute to their ability to cause invasive diseases in humans. PMID:26317436

  19. Epidemiological analysis of pneumococcal serotype 19A in healthy children following PCV7 vaccination.

    PubMed

    Tóthpál, A; Laub, K; Kardos, S; Tirczka, T; Kocsis, A; VAN DER Linden, M; Dobay, O

    2016-05-01

    After the introduction of conjugate vaccines, a strong rearrangement of pneumococcal serotypes was observed globally. Probably most concerning was the emergence of serotype 19A, which has not only high invasive disease potential, but also high antibiotic resistance. In the current study we focused on the increased prevalence of serotype 19A after the PCV vaccination rate became widely used in Hungary. A total of 2262 children aged 3-6 years were screened for pneumococcus carriage using nasal swabs. Children were divided into two groups according to the vaccination rates, low level (group 1) vs. high level (group 2). While the carriage rate did not change over time (average 32·9%), the serotype distribution differed greatly in the two groups. The prevalence of serotype 19A increased >eightfold. Almost all 19A isolates had high-level macrolide resistance and elevated penicillin minimum inhibitory concentrations. Genotyping methods revealed that these new 19A isolates are different from the previously frequent Hungary19A-6 PMEN clone. Both the carriage rate and the overall penicillin and macrolide resistance remained stable over time, but while several serotypes were represented in group 1, serotype 19A alone was clearly dominant in group 2. PMID:26548594

  20. El Niño-Southern Oscillation, local weather and occurrences of dengue virus serotypes

    NASA Astrophysics Data System (ADS)

    Huang, Xiaodong; Clements, Archie C. A.; Williams, Gail; Devine, Gregor; Tong, Shilu; Hu, Wenbiao

    2015-11-01

    Severe dengue fever is usually associated with secondary infection by a dengue virus (DENV) serotype (1 to 4) that is different to the serotype of the primary infection. Dengue outbreaks only occur following importations of DENV in Cairns, Australia. However, the majority of imported cases do not result in autochthonous transmission in Cairns. Although DENV transmission is strongly associated with the El Niño-Southern Oscillation (ENSO) climate cycle and local weather conditions, the frequency and potential risk factors of infections with the different DENV serotypes, including whether or not they differ, is unknown. This study used a classification tree model to identify the hierarchical interactions between Southern Oscillation Index (SOI), local weather factors, the presence of imported serotypes and the occurrence of the four autochthonous DENV serotypes from January 2000-December 2009 in Cairns. We found that the 12-week moving average of SOI and the 2-week moving average of maximum temperature were the most important factors influencing the variation in the weekly occurrence of the four DENV serotypes, the likelihoods of the occurrence of the four DENV serotypes may be unequal under the same environmental conditions, and occurrence may be influenced by changes in global and local environmental conditions in Cairns.