Mølbak, K.; Gerner-Smidt, P.; Wegener, Henrik Caspar
Until recently, Salmonella enterica serotype Enteritidis has remained sensitive to most antibiotics. However, national surveillance data from Denmark show that quinolone resistance in S. Enteritidis has increased from 0.8% in 1995 to 8.5% in 2000. These data support concerns that the current use...
Mølbak, K.; Gerner-Smidt, P.; Wegener, Henrik Caspar
Until recently, Salmonella enterica serotype Enteritidis has remained sensitive to most antibiotics. However, national surveillance data from Denmark show that quinolone resistance in S. Enteritidis has increased from 0.8% in 1995 to 8.5% in 2000. These data support concerns that the current use...... of quinolone in food animals leads to increasing resistance in S. Enteritidis and that action should be taken to limit such use....
Deng, Xiangyu; Desai, Prerak T.; den Bakker, Henk C.
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...
Madaczak, Grzegorz; Szych, Jolanta; Wasiak, Monika
Traditionally Salmonella enterica subsp. enterica serotypes are identified by slide agglutination with specific antisera for somatic, flagellar and sometimes capsular antigens. An alternative way is genoserotyping using for example a microarray, eg. commercially available test Check&Trace Salmonella. The goal of this study was to evaluate the Check&Trace Salmonella microarray for Salmonella enterica subsp. enterica serotype identification, using Salmonella strains provided by reference laboratories during External Quality Assurance Systems organized for national reference laboratories by ECDC and WHO GFN. 80 Salmonella enterica subsp. enterica have been tested using Check & Trace Salmonella (Check-Points BC, Netherlands). Also classical slide agglutination was performed according to EN ISO 6579:2003/Al:2007 norm, used as reference method. In the group of 80 tested strains, 66% were identified correctly, 4% gave uncertain results and 29% showed "Salmonella, genovar" without a serotype, of which 69% were not included in the CTS list of serotypes. Finally one strain has been recognized incorrectly. Because of IVD certification lack, the CTS test could not be recommended to clinical laboratories. AOAC-RI and OIE certification for test cause, that CTS could be used in most food, environmental and veterinary laboratories with the condition, that all unrecognized strains should be sent to a reference laboratory, to type according to EN ISO 6579:2003/Al:2007 norm, by KWM serotyping or other equal alternative methods.
Jackson, Brendan R; Griffin, Patricia M; Cole, Dana; Walsh, Kelly A; Chai, Shua J
Salmonella enterica infections are transmitted not only by animal-derived foods but also by vegetables, fruits, and other plant products. To clarify links between Salmonella serotypes and specific foods, we examined the diversity and predominance of food commodities implicated in outbreaks of salmonellosis during 1998-2008. More than 80% of outbreaks caused by serotypes Enteritidis, Heidelberg, and Hadar were attributed to eggs or poultry, whereas >50% of outbreaks caused by serotypes Javiana, Litchfield, Mbandaka, Muenchen, Poona, and Senftenberg were attributed to plant commodities. Serotypes Typhimurium and Newport were associated with a wide variety of food commodities. Knowledge about these associations can help guide outbreak investigations and control measures.
Griffin, Patricia M.; Cole, Dana; Walsh, Kelly A.; Chai, Shua J.
Salmonella enterica infections are transmitted not only by animal-derived foods but also by vegetables, fruits, and other plant products. To clarify links between Salmonella serotypes and specific foods, we examined the diversity and predominance of food commodities implicated in outbreaks of salmonellosis during 1998–2008. More than 80% of outbreaks caused by serotypes Enteritidis, Heidelberg, and Hadar were attributed to eggs or poultry, whereas >50% of outbreaks caused by serotypes Javiana, Litchfield, Mbandaka, Muenchen, Poona, and Senftenberg were attributed to plant commodities. Serotypes Typhimurium and Newport were associated with a wide variety of food commodities. Knowledge about these associations can help guide outbreak investigations and control measures. PMID:23876503
Cao, Guojie; Allard, Marc W; Hoffmann, Maria; Monday, Steven R; Muruvanda, Tim; Luo, Yan; Payne, Justin; Rump, Lydia; Meng, Kevin; Zhao, Shaohua; McDermott, Patrick F; Brown, Eric W; Meng, Jianghong
Multidrug-resistant (MDR) Salmonella enterica subsp. enterica serotype Newport has been a long-standing public health concern in the United States. We present the complete sequences of six IncA/C plasmids from animal-derived MDR S. Newport ranging from 80.1 to 158.5 kb. They shared a genetic backbone with S. Newport IncA/C plasmids pSN254 and pAM04528. Copyright © 2015 Cao et al.
Al Naiemi, Nashwan; Zwart, Bastiaan; Rijnsburger, Martine C.; Roosendaal, Robert; Debets-Ossenkopp, Yvette J.; Mulder, Janet A.; Fijen, Cees A.; Maten, Willemina; Vandenbroucke-Grauls, Christina M.; Savelkoul, Paul H.
A Salmonella enterica serotype Typhi strain was cultured from blood and fecal samples from a 54-year-old man with fever and diarrhea. He had returned from travel to the Philippines a few days earlier. Phenotypic and genotypic analysis confirmed the production of the SHV-12 extended-spectrum
Dr. Mike Miller reads an abridged version of the Emerging Infectious Diseasesâ study, Outbreak-associated Salmonella enterica Serotypes and Food Commodities, United States, 1998- 2008. Created: 9/9/2013 by National Center for Emerging and Zoonotic Infectious Diseases (NCEZID). Date Released: 9/9/2013.
Stevens, Marc J A; Zurfluh, Katrin; Althaus, Denise; Corti, Sabrina; Lehner, Angelika; Stephan, Roger
The genome of Salmonella enterica subsp. enterica serotype Senftenberg N17-509, a strain isolated from desiccated coconut, was sequenced using single-molecule real-time sequencing. It consists of a 5.1-Mbp chromosome and a 29-kb linear plasmid. Copyright © 2018 Stevens et al.
Olsen, J. E.; Skov, M. N.; Angen, Øystein
The genomic relationship between isolates representing 17 definitive phage types (DTs) of Salmonella enterica subsp. enterica serotype typhimurium (S. typhimurium) were analysed using three different typing methods: IS200 typing using the restriction enzymes EcoRI and Pvull, ribotyping using Smal...
Pulido-Landínez, M; Sánchez-Ingunza, R; Guard, J; Pinheiro do Nascimento, V
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. 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. Published 2013. This article is a U.S. Government work and is in the public domain in the USA
Full Text Available Salmonella enterica serotype Enteritidis is known as one of the most common pathogenic bacteria causing salmonellosis in humans. Raw materials of animal origin (eggs, chicken meat are frequent vectors that transmit this bacterium. Since eggs are used for the production of pasta, due to insufficient thermal treatment during pasta drying, they can be a potential risk to consumer health. Different essential oils of herbs can be used to reduce present pathogenic microorganisms. This paper compares a decrease in the number of Salmonella enterica serotype Enteritidis (D ATCC 13076 and Salmonella enterica serotype Enteritidis isolated from outbreaks of salmonellosis in egg-based pasta under the influence of thyme and sweet basil essential oils. The results indicate that the utilized oils were more effective against the epidemic strain than the ATCC strain. In addition, thyme oil caused a more significant inhibition of Salmonella enterica serotype Enteritidis during the production process.
Herman, Haley K; Hampshire, Karly N; Khoshnam, Nasim; Khan, Aleena A; Jerris, Robert; Abramowsky, Carlos R; Shehata, Bahig M
Bacterial infection of Salmonella enterica serotype Typhi is rare in the United States but endemic in many developing countries. Approximately 3-5% of patients become chronic asymptomatic carriers. We describe an atypical presentation of S. enterica serotype Typhi infection in a 10-year-old male, whose cholecystechtomy and bile culture revealed chronic carrier status despite negative stool tests and the absence of gallstones. The gallbladder showed marked thickening of the wall with an intense suppurative granulomatous reaction.
Whole genome analysis of foodborne pathogens is standard practice for regulatory agencies in the US, and a goal of 1 million genomes for Salmonella enterica is currently 76% complete. Inclusion of a range of Salmonella enterica subspecies I serotypes is important because only 30 of approximately 150...
Liebana, Ernesto; Batchelor, Miranda; Torres, Carmen; Briñas, Laura; Lagos, Luis A.; Abdalhamid, Baha; Hanson, Nancy D.; Martinez-Urtaza, Jaime
We report the case of a pediatric patient with a Salmonella enterica serotype Infantis infection. Detailed microbiological investigation revealed that this isolate carries four β-lactamase genes (blaTEM-1b variant, blaSHV-5, blaCTX-M-15, and blaCMY-2) conferring resistance to all β-lactams but imipenem. This is the first report of a Salmonella isolate with CTX-M and AmpC enzymes on the American continent, the first report of blaCMY-2 in Salmonella serotype Infantis, and the first report of blaCTX-M-15 in the genus Salmonella. PMID:15472372
Fardsanei, Fatemeh; Soltan Dallal, Mohammad Mehdi; Douraghi, Masoumeh; Zahraei Salehi, Taghi; Mahmoodi, Mahmood; Memariani, Hamed; Nikkhahi, Farhad
Salmonella enterica subspecies enterica serotype Enteritidis (S. Enteritidis) is one of the leading causes of food-borne gastroenteritis associated with the consumption of contaminated food products of animal origin. Little is known about the genetic diversity and virulence content of S. Enteritidis isolated from poultry meats and eggs in Iran. A total of 34 S. Enteritidis strains were collected from different food sources of animal origin in Tehran from May 2015 to July 2016. All of the S. Enteritidis strains were serotyped, antimicrobial susceptibility tested, and characterized for virulence genes. Pulsed-field gel electrophoresis (PFGE) was also applied for comparison of genetic relatedness. All of the strains harbored invA, hilA, ssrA, sefA, spvC, and sipA genes. A high prevalence of resistance against certain antibiotics such as cefuroxime (79.4%), nalidixic acid (47%), and ciprofloxacin (44.2%) was also observed. Regarding PFGE, S. Enteritidis strains from different sources showed considerable overlap, suggesting the lack of diversity among these isolates. Moreover, no correlation between virulence profiles or antibiotypes and PFGE clusters was observed. In conclusion, our study provided valuable information on virulence gene content, antibiotic resistance, and genetic diversity of S. Enteritidis isolated from food sources. Copyright © 2017 Elsevier Ltd. All rights reserved.
Achtman, Mark; Wain, John; Weill, François-Xavier; Nair, Satheesh; Zhou, Zhemin; Sangal, Vartul; Krauland, Mary G; Hale, James L; Harbottle, Heather; Uesbeck, Alexandra; Dougan, Gordon; Harrison, Lee H; Brisse, Sylvain
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.
Full Text Available 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.
Zhou, Zhemin; Sangal, Vartul; Krauland, Mary G.; Hale, James L.; Harbottle, Heather; Uesbeck, Alexandra; Dougan, Gordon; Harrison, Lee H.; Brisse, Sylvain
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
Tukel, Cagla; Akcelik, Mustafa; de Jong, Maarten F.; Simsek, Omer; Tsolis, Renee M.; Baumler, Andreas J.
MisL is a Salmonella enterica serotype Typhimurium fibronectin binding protein whose expression is induced during infection of mice. T-POP transposon mutagenesis identified marT as a positive regulatory element controlling expression of a misL::lacZYA transcriptional fusion. Gel shift analysis
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
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. Copyright © 2015 Elsevier B.V. and the International Society of Chemotherapy. All rights reserved.
Molbak, K.; Baggesen, Dorte Lau; Aarestrup, Frank Møller
Background Food-borne salmonella infections have become a major problem in industrialized countries. The strain of Salmonella enterica serotype typhimurium known as definitive phage type 104 (DT104) is usually resistant to five drugs: ampicillin, chloramphenicol, streptomycin, sulfonamides......, and tetracycline. An increasing proportion of DT104 isolates also have reduced susceptibility to fluoroquinolones. Methods The Danish salmonella surveillance program determines the phage types of all typhimurium strains from the food chain, and in the case of suspected outbreaks, five-drug-resistant strains...... are characterized by molecular methods. All patients infected with five-drug-resistant typhimurium are interviewed to obtain clinical and epidemiologic data. In 1998, an outbreak of salmonella occurred, in which the strain of typhimurium DT104 was new to Denmark. We investigated this outbreak and report our...
Full Text Available Salmonella enterica serovar Heidelberg is among the most detected serovars in swine and poultry, ranks among the top five serotypes associated with human salmonellosis and is disproportionately associated with invasive infections and mortality in humans. Salmonella are known to carry plasmids associated with antimicrobial resistance and virulence. To identify plasmid-associated genes in multidrug resistant S. enterica serovar Heidelberg, antimicrobial resistance plasmids from five isolates were sequenced using the 454 LifeSciences pyrosequencing technology. Four of the isolates contained incompatibility group (Inc A/C multidrug resistance plasmids harboring at least eight antimicrobial resistance genes. Each of these strains also carried a second resistance plasmid including two IncFIB, an IncHI2 and a plasmid lacking an identified Inc group. The fifth isolate contained an IncI1 plasmid, encoding resistance to gentamicin, streptomycin and sulfonamides. Some of the IncA/C plasmids lacked the full concert of transfer genes and yet were able to be conjugally transferred, likely due to the transfer genes carried on the companion plasmids in the strains. Several non-IncA/C resistance plasmids also carried putative virulence genes. When the sequences were compared to previously sequenced plasmids, it was found that while all plasmids demonstrated some similarity to other plasmids, they were unique, often due to differences in mobile genetic elements in the plasmids. Our study suggests that Salmonella Heidelberg isolates harbor plasmids that co-select for antimicrobial resistance and virulence, along with genes that can mediate the transfer of plasmids within and among other bacterial isolates. Prevalence of such plasmids can complicate efforts to control the spread of S. enterica serovar Heidelberg in food animal and human populations.
Lawrence Mark L
Full Text Available Abstract Background Salmonellosis may be a food safety problem when raw food products are mishandled and not fully cooked. In previous work, we developed bioluminescent Salmonella enterica serotypes using a plasmid-based reporting system that can be used for real-time monitoring of the pathogen's growth on food products in short term studies. In this study, we report the use of a Tn7-based transposon system for subcloning of luxCDABE genes into the chromosome of eleven Salmonella enterica serotypes isolated from the broiler production continuum. Results We found that the lux operon is constitutively expressed from the chromosome post-transposition and the lux cassette is stable without external pressure, i.e. antibiotic selection, for all Salmonella enterica serotypes used. Bioluminescence expression is based on an active electron transport chain and is directly related with metabolic activity. This relationship was quantified by measuring bioluminescence against a temperature gradient in aqueous solution using a luminometer. In addition, bioluminescent monitoring of two serotypes confirmed that our chicken skin model has the potential to be used to evaluate pathogen mitigation strategies. Conclusions This study demonstrated that our new stable reporting system eliminates bioluminescence variation due to plasmid instability and provides a reliable real-time experimental system to study application of preventive measures for Salmonella on food products in real-time for both short and long term studies.
Sheikh, Alaullah; Charles, Richelle C.; Rollins, Sean M.; Harris, Jason B.; Bhuiyan, Md. Saruar; Khanam, Farhana; Bukka, Archana; Kalsy, Anuj; Porwollik, Steffen; Brooks, W. Abdullah; LaRocque, Regina C.; Hohmann, Elizabeth L.; Cravioto, Alejandro; Logvinenko, Tanya; Calderwood, Stephen B.
Background: Salmonella enterica serotype Paratyphi A is a human-restricted cause of paratyphoid fever, accounting for up to a fifth of all cases of enteric fever in Asia. Methodology/Principal Findings: In this work, we applied an RNA analysis method, Selective Capture of Transcribed Sequences (SCOTS), and cDNA hybridization-microarray technology to identify S. Paratyphi A transcripts expressed by bacteria in the blood of three patients in Bangladesh. In total, we detected 1,798 S. Paratyphi ...
Kim, Jin Seok; Yun, Young-Sun; Kim, Soo Jin; Jeon, Se-Eun; Lee, Deog-Yong; Chung, Gyung Tae; Yoo, Cheon-Kwon; Kim, Junyoung
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.
Bogomolnaya Lydia M
Full Text Available Abstract 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.
Full Text Available 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.
Fabre, Laetitia; Le Hello, Simon; Roux, Chrystelle; Issenhuth-Jeanjean, Sylvie; Weill, François-Xavier
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
Kee Tai Goh
Full Text Available Introduction: This paper describes the epidemiological, microbiological and environmental investigations conducted during an outbreak of Salmonella gastroenteritis in Singapore.Methods: A case-control study was undertaken to identify the vehicle of transmission. Microbiological testing was performed on faecal, food and environmental samples. Isolates of Salmonella were further characterized by phage typing and ribotyping.Results: There were 216 gastroenteritis cases reported from 20 November to 4 December 2007. The causative agent was identified as Salmonella enterica subspecies enterica serotype Enteritidis for 14 out of 20 cases tested. The vehicle of transmission was traced to cream cakes produced by a bakery and sold at its retail outlets (P < 0.001, OR = 143.00, 95% Cl = 27.23–759.10. More than two-thirds of the 40 Salmonella strains isolated from hospitalized cases, food samples and asymptomatic food handlers were of phage type 1; the others reacted but did not conform to any phage type. The phage types correlated well with their unique antibiograms. The ribotype patterns of 22 selected isolates tested were highly similar, indicating genetic relatedness. The dendrogram of the strains from the outbreak showed distinct clustering and correlation compared to the non-outbreak strains, confirming a common source of infection.Discussion: The cream cakes were likely contaminated by one of the ingredients used in the icing. Cross-contamination down the production line and subsequent storage of cakes at ambient temperatures for a prolonged period before consumption could have resulted in the outbreak.
Weening, Eric H; Barker, Jared D; Laarakker, Marijke C; Humphries, Andrea D; Tsolis, Renée M; Bäumler, Andreas J
Salmonella enterica serotype Typhimurium causes human infections that can frequently be traced back through the food chain to healthy livestock whose intestine is colonized by the pathogen. Little is known about the genes important for intestinal carriage of S. enterica serotype Typhimurium in vertebrate animals. Here we characterized the role of 10 fimbrial operons, agf, fim, lpf, pef, bcf, stb, stc, std, stf, and sth, using competitive infection experiments performed in genetically susceptible (BALB/c) and resistant (CBA) mice. Deletion of agfAB, fimAICDHF, lpfABCDE, pefABCDI, bcfABCDEFG, stbABCD, stcABCD, stdAB, stfACDEFG, or sthABCDE did not reduce the ability of S. enterica serotype Typhimurium to colonize the spleen and cecum of BALB/c mice 5 days after infection. Similarly, deletion of agfAB, fimAICDHF, pefABCDI, and stfACDEFG did not result in reduced recovery of S. enterica serotype Typhimurium from fecal samples collected from infected CBA mice over a 30-day time period. However, S. enterica serotype Typhimurium strains carrying deletions in lpfABCDE, bcfABCDEFG, stbABCD, stcABCD, stdAB, or sthABCDE were recovered at significantly reduced numbers from the feces of CBA mice. There was a good correlation (R(2) = 0.9626) between competitive indices in the cecum and fecal samples of CBA mice at 30 days after infection, suggesting that the recovery of S. enterica serotype Typhimurium from fecal samples closely reflected its ability to colonize the cecum. Collectively, these data show that six fimbrial operons (lpf, bcf, stb, stc, std, and sth) contribute to long-term intestinal carriage of S. enterica serotype Typhimurium in genetically resistant mice.
Full Text Available 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. Results of phenotypic assays showed diversity of survival capabilities 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.
Full Text Available The main goal of this study was to establish how the inflammation caused by infection with two different Salmonella enterica serotypes, S. Typhimurium and S. Enteritidis, may lead to the predisposition to allergy as measured by total IgE level in the blood. Infection by S. Typhimurium did not affect the systemic IgE concentration while in S. Enteritidis-infected patients there was a significant 3.5-fold increase. This effect was especially profound in patients >4 years old, with up to the eight-fold increase above the norm. The degree of dysbiosis in these two infections measured with the comparative counts of cultivated bacteria showed an inverse relationship with the IgE concentration. Earlier we reported the elevated level of IL-17 in patients infected by S. Enteritidis. In the current study a significant correlation was found between the concentrations of IL-17 and IgE suggesting a possible role played by this cytokine in triggering the production of IgE in response to S. Enteritidis infection.
Full Text Available The benefits of probiotic bacteria have been widely explored. However, fermented foods and digestive system of humans and animals are an inexhaustible source of new potentially probiotic microorganisms. In this study we present three new Lactobacillus plantarum strains isolated from different dairy products: cow′s cheese, sheep′s cheese and whey. In order to determine the antibacterial activity of yet unexplored L. plantarum strains against Salmonella enterica serotype Typhimurium, in vitro competition and co-culture tests were done. Furthermore, adhesion of these strains to Caco-2 cells and their influence on the adhesion of Salmonella were tested. Results showed the potential probiotic activity of isolated strains. L. plantarum strains survived in the presence of 1 % bile salts, they possessed acidification ability, antibacterial activity and significantly attenuated the growth of S. Typhimurium in brain heart infusion broth. All tested L. plantarum strains were able to adhere to Caco-2 cells and significantly impair the adhesion of S. Typhimurium. All three L. plantarum strains exhibited signifi cant probiotic potential and anti-Salmonella activity; therefore, further testing on in vivo models should follow.
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.
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.
Simon eLe Hello
Full Text Available 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.
Yang, Wei-Chiun; Chan, Oi-Wa; Wu, Tsu-Lan; Chen, Chyi-Liang; Su, Lin-Hui; Chiu, Cheng-Hsun
The majority of nontyphoid Salmonella infection is identified in children. When an invasive or severe Salmonella infection is encountered, ceftriaxone is recommended for such patients. A 2-year-old girl was hospitalized for the treatment of Salmonella bacteremia and discharged with standard ceftriaxone treatment. She was readmitted to the hospital after 2 days due to the recurrence of the Salmonella bacteremia. The study aimed to unveil the mechanism for the relapse. Six isolates (4 blood and 2 stool) were recovered from the patient, with the last two blood isolates being ceftriaxone-resistant. Pulsed-field gel electrophoresis was used for genotyping. Ceftriaxone resistance genes and transferability of the resistance plasmid were examined by molecular methods. All isolates were identified as Salmonella enterica serotype Oranienburg. Five isolates demonstrated almost identical electrophoresis patterns, except that in the two ceftriaxone-resistant isolates an extra band (>100 kb) was noted. A blaCMY-2 gene, carried by a 120-kb conjugative IncI1 plasmid of the sequence type 53, was identified in the two ceftriaxone-resistant isolates. Transfer of the resistance plasmid from one blood isolate to Escherichia coli J53 resulted in the increase of ceftriaxone minimum inhibitory concentration from 0.125 μg/mL to 32 μg/mL in the recipient. Ceftriaxone is the standard therapeutic choice for invasive or serious Salmonella infections in children. Pediatricians should be aware of the possibility of resistance development during therapy, especially in areas with a widespread of ceftriaxone resistance genes that are carried by a self-transferrable plasmid, such as the blaCMY-2-carrying IncI1 plasmid identified herein. Copyright © 2014. Published by Elsevier B.V.
Ng, Kamela Charmaine S; Rivera, Windell L
Food poisoning outbreaks and livestock mortalities caused by Salmonella enterica are widespread in the Philippines, with hogs being the most commonly recognized carriers of the pathogen. To prevent and control the occurrence of S. enterica infection in the country, methods were used in this study to isolate and rapidly detect, differentiate, and characterize S. enterica in tonsils and jejuna with jejunal lymph nodes of swine slaughtered in four locally registered meat establishments (LRMEs) and four meat establishments accredited by the National Meat Inspection Services in Metro Manila. A total of 480 samples were collected from 240 animals (120 pigs from each type of meat establishment). A significantly higher proportion of pigs were positive for S. enterica in LRMEs (60 of 120) compared with meat establishments accredited by the National Meat Inspection Services (38 of 120). More S. enterica-positive samples were found in tonsils compared with jejuna with jejunal lymph nodes in LRMEs, but this difference was not significant. A PCR assay targeting the invA gene had sensitivity that was statistically similar to that of the culture method, detecting 93 of 98 culture-confirmed samples. Multiplex PCR-based O-serogrouping and H/Sdf I typing revealed four S. enterica serogroups (B, C1, D, and E) and six serotypes (Agona, Choleraesuis, Enteritidis, Heidelberg, Typhimurium, and Weltevreden), respectively, which was confirmed by DNA sequencing of the PCR products. This study was the first to report detection of S. enterica serotype Agona in the country.
Full Text Available Deepa Harichandran, Kavitha Radhakrishnan Dinesh Department of Microbiology, Amrita School of Medicine, Amrita Institute of Medical Sciences and Research Centre, Amrita Vishwa Vidyapeetham University, Kochi, Kerala, India Background/purpose: Typhoid and paratyphoid fever continue to be important causes of illness and death in parts of Asia, being associated with poor sanitation and consumption of unsafe food and water. Antimicrobial resistance has emerged to traditional first-line drugs, namely, the fluoroquinolones, as well as to third-generation cephalosporins, posing challenges to treatment. Azithromycin has proven to be an effective alternative for treatment of uncomplicated typhoid fever. The purpose of this study was to determine the antimicrobial susceptibility, clinical outcome and serotype distribution pattern of clinical isolates belonging to Salmonella enterica subspecies enterica. Methodology: All clinical isolates of S. enterica obtained from blood, sterile body fluids, as well as stool and urine samples at Amrita Institute of Medical Sciences and Research Centre, Kerala, India, between August 2011 and July 2013 were included in the study and processed based on standard microbiology protocols. Results: A total of 118 isolates of Salmonella were obtained during the study period. Out of these, 79 were of S. Typhi (66.95%, followed by isolates of S. Paratyphi A (22; 18.64% and S. Typhimurium 12 (10.17%. Five isolates could not be identified further. There was 100% susceptibility to ceftriaxone in all S. enterica subspecies. Ciprofloxacin susceptibility was 32.91% for S. Typhi and 40.90% for S. Paratyphi A as determined by the disk diffusion method. The susceptibility profile of S. Typhi isolates to different antimicrobials was as follows: chloramphenicol (94.93%, ampicillin (77.21%, cotrimoxazole (75.94% and azithromycin (78.48%. For S. Typhi, the minimum inhibitory concentration (MIC of ciprofloxacin required to inhibit the growth
Ranjbar, Reza; Ahmadi, Mitra; Memariani, Mojtaba
Salmonella is an important cause of food-borne infection worldwide. Detection of outbreaks caused by Salmonella spp. relies on suitable and robust methods for genotyping. Little is known about the genetic diversity of the Salmonella enterica subspecies enterica serotype Infantis strains isolated from human sources in Iran. In this study, 40 isolates of S. Infantis, which were previously recovered from patients with gastroenteritis or diarrhea in Tehran between years 2007 and 2009, were subjected to multiple-locus variable-number of tandem repeat (VNTR) analysis (MLVA), pulsed-field gel electrophoresis (PFGE), and ERIC-PCR. Using MLVA method, 31 types were identified. The MLVA clustering of the isolates by the unweighted pair group method with arithmetic mean (UPGMA) revealed the presence of two major clusters. The discriminatory power of MLVA was superior to that of PFGE and ERIC-PCR. Overall, our data showed that MLVA assay could effectively differentiate closely related strains. It is technically simple and inexpensive to perform. Furthermore, MLVA can be used as a helpful method for epidemiological investigations. Copyright © 2016 Elsevier Ltd. All rights reserved.
Aabo, Søren; Christensen, J.P.; Chadfield, M.S.
A. Two serotypes demonstrated intracellular log(10) counts that differed significantly from all other serotypes tested: Salmonella Enteritidis PT4 being 1.5 log(10) colony forming units (CFU) ( 31-fold) higher, and Salmonella Tennessee being 0.7 log(10) CFU (fivefold) lower than the reference strain (P...
Le Hello, Simon; Bekhit, Amany; Granier, Sophie A.
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......-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......A (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...
Vieira, A R; Grass, J; Fedorka-Cray, P J; Plumblee, J R; Tate, H; Cole, D J
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 (Salmonella Hadar.
Trebichavský, Ilja; Šplíchal, Igor; Šplíchalová, Alla; Muneta, Y.; Mori, Y.
Roč. 48, č. 3 (2003), s. 403-407 ISSN 0015-5632 R&D Projects: GA ČR GA524/01/0917; GA ČR GA524/02/1217 Institutional research plan: CEZ:AV0Z5020903 Keywords : salmonella * enterica Subject RIV: EE - Microbiology, Virology Impact factor: 0.857, year: 2003
Aabo, Søren; Christensen, J.P.; Chadfield, M.S.
The aim of the present study was to compare the invasion of selected zoonotic Salmonella serotypes of poultry in an in vivo chicken intestinal loop model and also in vitro in epithelial cell cultures. Invasion was measured relative to a reference strain, Salmonella Typhimurium 4/74 invH201::Tnpho......A. Two serotypes demonstrated intracellular log(10) counts that differed significantly from all other serotypes tested: Salmonella Enteritidis PT4 being 1.5 log(10) colony forming units (CFU) ( 31-fold) higher, and Salmonella Tennessee being 0.7 log(10) CFU (fivefold) lower than the reference strain (P......, S. Enteritidis PT6, S. Enteritidis PT8, and Salmonella Berta. The serotypes Salmonella Hadar, Salmonella Virchow, S. 4,12: b:-, S. Typhimurium DT41, and Salmonella Infantis, most of which are considered horizontally transmitted, did not show significantly different intracellular counts from...
Thomas R. Connor
Full Text Available For 100 years, it has been obvious that Salmonella enterica strains sharing the serotype with the formula 1,4,,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.
Sola, Christophe; Abadia, Edgar; Le Hello, Simon; Weill, François-Xavier
International audience; Spoligotyping was developed almost 18 years ago and still remains a popular fi rst-lane genotyping technique to identify and subtype Mycobacterium tuberculosis complex (MTC) clinical isolates at a phylogeo-graphic level. For other pathogens, such as Salmonella enterica , recent studies suggest that specifi cally designed spoligotyping techniques could be interesting for public health purposes. Spoligotyping was in its original format a reverse line-blot hybridization m...
Achtman, Mark; Wain, John; Weill, François-Xavier; Nair, Satheesh; Zhou, Zhemin; Sangal, Vartul; Krauland, Mary G.; Hales, James; Harbottle, Heather; Uesbeck, Alexandra; Dougan, Gordon; Harrison, Lee H.; Brisse, Sylvain
The co-authors included in the S. enterica MLST study group consist of : Derek Brown (Scottish Salmonella Reference Laboratory, Glasgow, UK), Edmundo Calva (Instituto de Biotecnología, Universidad Nacional Autónoma de México, Cuernavaca, Mexico), Chishih Chu (Department of Microbiology and Immunology, National Chiayi University, Chiayi,Taiwan), Martin Cormican (Department of Bacteriology and National Salmonella Reference Laboratory (GUH), National University of Ireland, Galway, Ireland), Xavi...
Kisluk, Guy; Yaron, Sima
Salmonella enterica is one of the major food-borne pathogens associated with ready-to-eat fresh foods. Although polluted water might be a significant source of contamination in the field, factors that influence the transfer of Salmonella from water to the crops are not well understood, especially under conditions of low pathogen levels in water. The aim of this study was to investigate the short- and long-term (1 h to 28 days) persistence of Salmonella enterica serotype Typhimurium in the phyllosphere and the rhizosphere of parsley following spray irrigation with contaminated water. Plate counting and quantitative real-time PCR (qRT-PCR)-based methods were implemented for the quantification. By applying qRT-PCR with enrichment, we were able to show that even irrigation with water containing as little as ∼300 CFU/ml resulted in the persistence of S. Typhimurium on the plants for 48 h. Irrigation with water containing 8.5 log CFU/ml resulted in persistence of the bacteria in the phyllosphere and the rhizosphere for at least 4 weeks, but the population steadily declined with a major reduction in bacterial counts, of ∼2 log CFU/g, during the first 2 days. Higher levels of Salmonella were detected in the phyllosphere when plants were irrigated during the night compared to irrigation during the morning and during winter compared to the other seasons. Further elucidation of the mechanisms underlying the transfer of Salmonella from contaminated water to crops, as well as its persistence over time, will enable the implementation of effective irrigation and control strategies.
Full Text Available Abstract Background Salmonella enterica subsp. enterica serotype Enteritidis is known as an important and pathogenic clonal group which continues to cause worldwide sporadic cases and outbreaks in humans. Here a new multiple-locus variable-number tandem repeat analysis (MLVA method is reported for highly-discriminative subtyping of Salmonella Enteritidis. Emphasis was given on the most predominant phage types PT4 and PT8. The method comprises multiplex PCR specifically amplifying repeated sequences from nine different loci followed by an automatic fragment size analysis using a multicolor capillary electrophoresis instrument. A total of 240 human, animal, food and environmental isolates of S. Enteritidis including 23 definite phage types were used for development and validation. Furthermore, the MLVA types were compared to the phage types of several isolates from two recent outbreaks to determine the concordance between both methods and to estimate their in vivo stability. The in vitro stability of the two MLVA types specifically for PT4 and PT8 strains were determined by multiple freeze-thaw cycles. Results Seventy-nine different MLVA types were identified in 240 S. Enteritidis strains. The Simpson's diversity index for the MLVA method was 0.919 and Nei diversity values for the nine VNTR loci ranged from 0.07 to 0.65. Twenty-four MLVA types could be assigned to 62 PT4 strains and 21 types to 81 PT8 strains. All outbreak isolates had an indistinguishable outbreak specific MLVA type. The in vitro stability experiments showed no changes of the MLVA type compared to the original isolate. Conclusion This MLVA method is useful to discriminate S. Enteritidis strains even within a single phage type. It is easy in use, fast, and cheap compared to other high-resolution molecular methods and therefore an important tool for surveillance and outbreak studies for S. Enteritidis.
Optical detection of foodborne bacteria such as Salmonella classifies bacteria by analyzing 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 spect...
Ben Salem, Rakia; Abbassi, Mohamed S; García, Vanesa; García-Fierro, Raquel; Fernández, Javier; Kilani, Hajer; Jaouani, Imen; Khayeche, Monia; Messadi, Lilia; Rodicio, María R
This study focused on 77 isolates of Salmonella enterica serotype Enteritidis collected during 2009 to 2013 from healthy and sick chickens and environmental farm samples in Tunisia. Resistance to 14 antimicrobials and the encoding genes were analyzed. 66, 26, 6.5, 3.9 and 1.3% were pan-susceptible or showed resistance to nalidixic acid (Asp87 to Tyr and Asp87 to Asn substitutions in GyrA), ampicillin (bla TEM-1-like and bla SHV ), sulfonamides (sul1and sul3) and streptomycin (strB), respectively. A single isolate with intermediate susceptibility to ciprofloxacin was positive for qnrB, whereas qnrA, qnrS or aac(6')-Ib-cr were not detected. The virulotype of the isolates was established by testing ten virulence genes. The orgA, ssaQ, mgtC, siiD, sopB genes, located on Salmonella pathogenicity islands, and spvC of the serotype-specific virulence plasmid, were common to all isolates. In contrast, the prophage-associated sopE-1, sodC1 and gipA genes and the fimbrial bcfC gene were variably represented. All isolates except one contained the virulence plasmid, which appeared either alone or together with one or more additional plasmids. One isolate carried a single plasmid of ca. 90Kb which may be derived from the virulence plasmid (60Kb). Overall, seven resistotypes, six virulotypes and six plasmid profiles were identified. XbaI-PFGE revealed four related pulsotypes (X1-X4), with 80% of the isolates sharing the X1 pattern. The latter isolates exhibited different resistance, virulence and plasmid profiles, suggesting that mobile genetic elements, particularly prophages and plasmids, are of central importance for the evolution and adaptation of S. Enteritidis circulating in chicken farms in Tunisia. Copyright © 2017 The Authors. Published by Elsevier Ltd.. All rights reserved.
Castro, M F P M; Rezende, A C B; Benato, E A; Valentini, S R T; Furlani, R P Z; Tfouni, S A V
The effect of phosphine on Salmonella enterica serotype Enteritidis inoculated in culture medium and in black pepper grains (Piper nigrum), as well as on the reduction of the microbial load of the dried and moisturized product, was verified. The postfumigation effect was verified in inoculated samples with 0.92 and 0.97 water activity (a(w)) exposed to 6 g/m(3) phosphine for 72 h, dried to 0.67 a(w), and stored for 24, 48, and 72 h. No decreases were observed in Salmonella Enteritidis populations in culture medium when fumigant concentrations up to 6 g/m(3) were applied for 48 h at 35°C. However, the colonies showed reductions in size and atypical coloration as the phosphine concentration increased. No reduction in Salmonella counts occurred on the inoculated dried samples after fumigation. On the other hand, when phosphine at concentrations of 6 g/m(3) was applied on moisturized black pepper for 72 h, decreases in Salmonella counts of around 80% were observed. The counts of total aerobic mesophilic bacterium populations of the dried and moisturized black pepper were not affected by the fumigant treatment. The results of the postfumigation studies indicated that Salmonella Enteritidis was absent in the fumigated grains after drying and storage for 72 h, indicating a promising application for this technique. It was concluded that for Salmonella Enteritidis control, phosphine fumigation could be applied to black pepper grains before drying and the producers should rigidly follow good agricultural practices, mainly during the drying process, in order to avoid product recontamination. Additional work is needed to confirm the findings with more Salmonella serotypes and strains.
Logan Julie MJ
Full Text Available Abstract Background The fliC and fljB genes in Salmonella code for the phase 1 (H1 and phase 2 (H2 flagellin respectively, the rfb cluster encodes the majority of enzymes for polysaccharide (O antigen biosynthesis, together they determine the antigenic profile by which Salmonella are identified. Sequencing and characterisation of fliC was performed in the development of a molecular serotyping technique. Results FliC sequencing of 106 strains revealed two groups; the g-complex included those exhibiting "g" or "m,t" antigenic factors, and the non-g strains which formed a second more diverse group. Variation in fliC was characterised and sero-specific motifs identified. Furthermore, it was possible to identify differences in certain H antigens that are not detected by traditional serotyping. A rapid short sequencing assay was developed to target serotype-specific sequence motifs in fliC. The assay was evaluated for identification of H1 antigens with a panel of 55 strains. Conclusion FliC sequences were obtained for more than 100 strains comprising 29 different H1 alleles. Unique pyrosequencing profiles corresponding to the H1 component of the serotype were generated reproducibly for the 23 alleles represented in the evaluation panel. Short read sequence assays can now be used to identify fliC alleles in approximately 97% of the 50 medically most important Salmonella in England and Wales. Capability for high throughput testing and automation give these assays considerable advantages over traditional methods.
Ferrato, Christina; Chui, Linda; King, Robin; Louie, Marie
Salmonella is one of the most common enteric pathogens related to foodborne illness. Alberta's Provincial Laboratory for Public Health (ProvLab) provides Outbreak and Surveillance support by performing serotyping. The Check&Trace Salmonella™ (CTS) assay (Check-Points, Netherlands), a commercial DNA microarray system, can determine the serotype designation of a Salmonella isolate with automated interpretation. Here we evaluate 1028 Salmonella isolates of human clinical or environmental sources in Alberta, Canada with the CTS assay. CTS was able to assign a serovar to 98.7% of the most frequently occurring human clinical strains in Alberta (82.5% overall), and 71.7% of isolates which were inconclusive by conventional methods. There was 99.7% concordance in environmental isolates. The CTS database has potential to expand to identify rare serovars. With the anticipated shift to molecular methods for identification, CTS provides an easy transition and demonstrates ease-of-use and reduces the turn-around-time of a reported result significantly compared to classical serotyping. Copyright © 2017 Elsevier B.V. All rights reserved.
Full Text Available In recent years, Salmonella enterica serovar Enteritidis has been a primary cause of human salmonellosis in many countries. The major objective of this study was to investigate genetic diversity among Salmonella Enteritidis strains from different origins (food and human by Enterobacterial Repetitive Intergenic Consensus (ERIC -PCR, as well as to assess their plasmid profiling and antimicrobial resistance. A total of 30 Salmonella Enteritidis isolates, 15 from food samples (chicken, lamb, beef and duck meats and 15 from clinical samples were collected in Tehran. Identification of isolates as Salmonella was confirmed by using conventional standard biochemical and serological tests. Multiplex-PCR was used for serotyping of isolates to identify Salmonella Enteritidis. Antimicrobial susceptibility testing to 16 agents founds drug resistance patterns among Salmonella Enteritidis isolates. No resistance was observed to cephalexin, ceftriaxone, ceftazidime and cefotaxime, ciprofloxacin, imipenem or meropenem, chloramphenicol and gentamicin. The highest resistance (96.7% was observed to nitrofurantoin. Seven plasmid profiles (P1–P7 were detected, and a 68-kb plasmid was found in all isolates. Two different primers; ERIC and (GTG5 were used for genotyping, which each produced four profiles. The majority of clinical and food isolates fell into two separate common types (CTs with a similar percentage of 95% by ERIC-PCR. Using primer (GTG5, 29 isolates incorporated in three CTs with 70% of isolates showing a single banding pattern. Limited genetic diversity among human and food isolates of Salmonella Enteritidis may indicate that contaminated foods were possibly the source of human salmonellosis. These results confirmed that ERIC-PCR genotyping has limited discriminatory power for Salmonella Enteritidis of different origin.
Chessa, Daniela; Winter, Maria G; Jakomin, Marcello; Bäumler, Andreas J
The std operon encodes a fimbrial adhesin of Salmonella enterica serotype Typhimurium that is required for attachment to intestinal epithelial cells and for cecal colonization in the mouse. To study the mechanism by which this virulence factor contributes to colonization we characterized its binding specificity. Std-mediated binding to human colonic epithelial (Caco-2) cells could be abrogated by removing N-linked glycans. Adherence of Std fimbriated S. Typhimurium to Caco-2 cells could be blocked by co-incubation with H type 2 oligosaccharide (Fucalpha1-2Galbeta1-4GlcNAc) or by pretreatment of cells with alpha1-2 fucosidase. In contrast, pretreatment of Caco-2 cells with neuraminidase or co-incubation with the type 2 disaccharide precursor (Galbeta1-4GlcNAc) did not reduce adherence of Std fimbriated S. Typhimurium. Binding of purified Std fimbriae to Fucalpha1-2Galbeta1-4GlcNAc in a solid phase binding assay was competitively inhibited by Ulex europaeus agglutinin-I (UEA-I), a lectin specific for Fucalpha1-2 moieties. Purified Std fimbriae and UEA both bound to a receptor localized in the mucus layer of the murine cecum. These data suggest that the std operon encodes an adhesin that binds an alpha1-2 fucosylated receptor(s) present in the cecal mucosa.
Delsol, Anne A; Woodward, Martin J; Roe, John M
There are concerns that the use of enrofloxacin in livestock production may contribute to the development of fluoroquinolone resistance in zoonotic bacteria. The objective of our study was to investigate the effect of a single 5 day enrofloxacin treatment on Salmonella enterica serotype Typhimurium DT104 in a pig model. Our results showed that a single treatment failed to eradicate S. Typhimurium DT104, which continued to be isolated up to 35 days after treatment. We also provide evidence that treatment positively selects for S. Typhimurium DT104 strains that are already nalidixic acid resistant (gyrA Asn-87) or cyclohexane resistant, the latter being indicative of an up-regulated efflux pump. Emergence of fluoroquinolone resistance was not detected during treatment or post-treatment in any of the Salmonella strains monitored. However, the effect of enrofloxacin on the nalidixic acid-resistant and cyclohexane-resistant S. Typhimurium DT104 outlasted the current withdrawal time of 10 days for Baytril (commercial veterinary formulation of enrofloxacin). In conclusion, our study has provided direct evidence that enrofloxacin-treated pigs could be entering abattoirs with higher numbers of quinolone-resistant zoonotic bacteria than untreated pigs, increasing the risk of these entering the food chain.
Irfan Ahmad Mir
Conclusions: Occurrence of high proportion of serovars in our study which can cause serious gastroenteritis in humans is a matter of concern. Salmonella Altona has been detected for the first time in India from poultry. This serotype is known to cause serious outbreaks of gastroenteritis in humans. Multidrug resistant isolates were recovered at high percentage which can be attributed to non-judicious use of antibiotics both in prophylaxis and treatment regimen. This observation draws serious attention as poultry serves as an important source of transmission of these multidrug resistant Salmonella serovars to humans.
Pantip, Sirichote; Aroon, B.; Kanokwan, Tienmanee
. 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......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...
Richelle C Charles
Full Text Available Salmonella enterica serotype Typhi can colonize and persist in the biliary tract of infected individuals, resulting in a state of asymptomatic chronic carriage. Chronic carriers may act as persistent reservoirs of infection within a community and may introduce infection to susceptible individuals and new communities. Little is known about the interaction between the host and pathogen in the biliary tract of chronic carriers, and there is currently no reliable diagnostic assay to identify asymptomatic S. Typhi carriage.To study host-pathogen interactions in the biliary tract during S. Typhi carriage, we applied an immunoscreening technique called in vivo-induced antigen technology (IVIAT, to identify potential biomarkers unique to carriers. IVIAT identifies humorally immunogenic bacterial antigens expressed uniquely in the in vivo environment, and we hypothesized that S. Typhi surviving in the biliary tract of humans may express a distinct antigenic profile. Thirteen S. Typhi antigens that were immunoreactive in carriers, but not in healthy individuals from a typhoid endemic area, were identified. The identified antigens included a number of putative membrane proteins, lipoproteins, and hemolysin-related proteins. YncE (STY1479, an uncharacterized protein with an ATP-binding motif, gave prominent responses in our screen. The response to YncE in patients whose biliary tract contained S. Typhi was compared to responses in patients whose biliary tract did not contain S. Typhi, patients with acute typhoid fever, and healthy controls residing in a typhoid endemic area. Seven of 10 (70% chronic carriers, 0 of 8 bile culture-negative controls (0%, 0 of 8 healthy Bangladeshis (0%, and 1 of 8 (12.5% Bangladeshis with acute typhoid fever had detectable anti-YncE IgG in blood. IgA responses were also present.Further evaluation of YncE and other antigens identified by IVIAT could lead to the development of improved diagnostic assays to identify asymptomatic
Full Text Available Salmonella enterica serotype Typhi is the cause of typhoid fever. It is a human-restricted pathogen, and few data exist on S. Typhi gene expression in humans.We applied an RNA capture and amplification technique, Selective Capture of Transcribed Sequences (SCOTS, and microarray hybridization to identify S. Typhi transcripts expressed in the blood of five humans infected with S. Typhi in Bangladesh. In total, we detected the expression of mRNAs for 2,046 S. Typhi genes (44% of the S. Typhi genome in human blood; expression of 912 genes was detected in all 5 patients, and expression of 1,100 genes was detected in 4 or more patients. Identified transcripts were associated with the virulence-associated PhoP regulon, Salmonella pathogenicity islands, the use of alternative carbon and energy sources, synthesis and transport of iron, thiamine, and biotin, and resistance to antimicrobial peptides and oxidative stress. The most highly represented group were genes currently annotated as encoding proteins designated as hypothetical, unknown, or unclassified. Of the 2,046 detected transcripts, 1,320 (29% of the S. Typhi genome had significantly different levels of detection in human blood compared to in vitro cultures; detection of 141 transcripts was significantly different in all 5 patients, and detection of 331 transcripts varied in at least 4 patients. These mRNAs encode proteins of unknown function, those involved in energy metabolism, transport and binding, cell envelope, cellular processes, and pathogenesis. We confirmed increased expression of a subset of identified mRNAs by quantitative-PCR.We report the first characterization of bacterial transcriptional profiles in the blood of patients with typhoid fever. S. Typhi is an important global pathogen whose restricted host range has greatly inhibited laboratory studies. Our results suggest that S. Typhi uses a largely uncharacterized genetic repertoire to survive within cells and utilize alternate
Mark R Wilson
Full Text Available 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
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
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
Brown, D. J.; Baggesen, Dorte Lau; Platt, D. J.
The plasmid pOG670, a 54 kb, conjugative plasmid that specifies resistance to ampicillin and kanamycin and belonging to the incompatibility group X (IncX), was transferred into 10 isolates of Salmonella enterica serotype Enteritidis belonging to 10 different phage types (PT1, 2, 3, 4, 8, 9, 9b, 10......, 6 and 13a respectively. The results indicate a different route for phage type conversion Enteritidis from others reported in the literature and, although IncX plasmids are not normally present in PT8 or PT13a, may suggest a possible mechanism/link connecting these phage types....
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
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.
Parvathi, A.; Vijayan, J.; Murali, G.; Chandran, P.
Salmonella enterica serotype Newport is an important cause of non-typhoidal salmonellosis, a clinically less severe infection than typhoid fever caused by S. enterica serotype Typhi. In this investigation, the virulence genotypes of S. enterica...
Full Text Available Abstract Background Salmonellosis is one of the most important foodborne diseases and a major threat to public health. Salmonella serotype Virchow ranks among the top five serovars in Europe. Method A total of 153 strains isolated from different patients from 2004 through 2009 in Switzerland were further characterized by (i assessing phenotypic antibiotic resistance profiles using the disk diffusion method and (ii by genotyping using pulsed-field gel electrophoresis (PFGE after macrorestriction with XbaI in order to evaluate strain relationship. Results The relative frequency of S. Virchow among other Salmonella serovars varied between 4th to 8th rank. The annual incidence ranged from 0.45/100'000 in 2004 to 0.40/100'000 in 2009. A total of 48 strains (32% were resistant to one to 3 antimicrobials, 54 strains (36% displayed resistance patterns to more than three antibiotics. No trend was identifiable over the years 2004 to 2009. We found a high prevalence (62% of nalidixic acid resistant strains, suggesting an equally high rate of decreased fluoroqionolone susceptibility, whereas intermediate resistance to ciprofloxacin was negligible. Two strains were extended spectrum β-lactamase (ESBL producers. Analysis of PFGE patterns uncovered a predominant cluster (similarity coefficient above 80% consisting of 104 of the 153 strains. Conclusion The worldwide increase of antibiotic resistances in Salmonella is an emerging public health problem. For Switzerland, no clear trend is identifiable over the years 2004 to 2009 for S. Virchow. Antimicrobial susceptibility and resistance profiles varied considerably within this period. Nevertheless, the situation in Switzerland coincided with findings in other European countries. Genotyping results of this strain collection revealed no evidence for an undetected outbreak within this time period.
Chadfield, M. S.; Christensen, J. P.; Madsen, Mogens
An increased prevalence of Salmonella enterica serovar Tennessee (6, 7: z(29):-) was observed in broiler flocks in Denmark in 1994 and a parallel increase in the prevalence of Salmonella enterica serovar 6, 7:-:- was demonstrated, albeit at a lower level. Plasmid profiling and ribotyping revealed...
Full Text Available Salmonella enterica serotype Paratyphi A is a human-restricted cause of paratyphoid fever, accounting for up to a fifth of all cases of enteric fever in Asia.In this work, we applied an RNA analysis method, Selective Capture of Transcribed Sequences (SCOTS, and cDNA hybridization-microarray technology to identify S. Paratyphi A transcripts expressed by bacteria in the blood of three patients in Bangladesh. In total, we detected 1,798 S. Paratyphi A mRNAs expressed in the blood of infected humans (43.9% of the ORFeome. Of these, we identified 868 in at least two patients, and 315 in all three patients. S. Paratyphi A transcripts identified in at least two patients encode proteins involved in energy metabolism, nutrient and iron acquisition, vitamin biosynthesis, stress responses, oxidative stress resistance, and pathogenesis. A number of detected transcripts are expressed from PhoP and SlyA-regulated genes associated with intra-macrophage survival, genes contained within Salmonella Pathogenicity Islands (SPIs 1-4, 6, 10, 13, and 16, as well as RpoS-regulated genes. The largest category of identified transcripts is that of encoding proteins with unknown function. When comparing levels of bacterial mRNA using in vivo samples collected from infected patients to samples from in vitro grown organisms, we found significant differences for 347, 391, and 456 S. Paratyphi A transcripts in each of three individual patients (approximately 9.7% of the ORFeome. Of these, expression of 194 transcripts (4.7% of ORFs was concordant in two or more patients, and 41 in all patients. Genes encoding these transcripts are contained within SPI-1, 3, 6 and 10, PhoP-regulated genes, involved in energy metabolism, nutrient acquisition, drug resistance, or uncharacterized genes. Using quantitative RT-PCR, we confirmed increased gene expression in vivo for a subset of these genes.To our knowledge, we describe the first microarray-based transcriptional analysis of a pathogen
Bartkus Joanne M
Full Text Available Abstract Background Salmonella enterica serotype Enteritidis (S. Enteritidis is a zoonotic pathogen, which can be found in many sources including animals and the environment. However, little is known about the molecular relatedness among S. Enteritidis isolates from different sources. We have applied multiple-locus variable number tandem repeat analysis (MLVA to study the genetic diversity of S. Enteritidis isolates from human and non-human sources. Results We identified 38 unique MLVA types using nine VNTR loci markers for discrimination between 145 S. Enteritidis isolates from different sources including humans (n = 41, chickens (n = 45, and eggs (n = 40. There were 20 distinct MLVA types identified from human isolates, 17 distinct MLVA types from chicken isolates, and 5 from egg isolates. We compared allele distribution and frequency for each VNTR marker and measured allelic polymorphism within each VNTR locus of S. Enteritidis isolates from the sources using Nei's diversity index (D. Differences in allele distribution and frequency were detected in most loci of study isolates. Different genetic diversity for certain loci was identified in isolates from different sources. The average of genetic diversity (D was lower in egg isolates (0.16 compared to human (0.41 and chicken (0.30. However, for loci SE3, SE7, and SE9, human isolates showed significantly higher diversity than both chicken and egg isolates. Whereas for loci SE5 and SE10, chicken isolates had significantly higher diversity than both human and egg isolates. Minimum-spanning tree (MST comprised one major cluster, a minor cluster, and four clonal expansions. MLVA application enabled a cluster analysis by the MST of the S. Enteritidis isolates by sources, which allows a great insight into the genetic relatedness and the possible flow of these organisms between different reservoirs and humans. Conclusion Differences in allele distribution and genetic diversity of VNTR loci in S
Mohamed A. El-Tayeb
Full Text Available Abstract Salmonella is recognized as a common foodborne pathogen, causing major health problems in Saudi Arabia. Herein, we report epidemiology, antimicrobial susceptibility and the genetic basis of resistance among S. enterica strains isolated in Saudi Arabia. Isolation of Salmonella spp. from clinical and environmental samples resulted in isolation of 33 strains identified as S. enterica based on their biochemical characteristics and 16S-rDNA sequences. S. enterica serovar Enteritidis showed highest prevalence (39.4%, followed by S. Paratyphi (21.2%, S. Typhimurium (15.2%, S. Typhi and S. Arizona (12.1%, respectively. Most isolates were resistant to 1st and 2nd generation cephalosporin; and aminoglycosides. Moreover, several S. enterica isolates exhibited resistance to the first-line antibiotics used for Salmonellosis treatment including ampicillin, trimethoprim-sulfamethoxazole and chloramphenicol. In addition, the results revealed the emergence of two S. enterica isolates showing resistance to third-generation cephalosporin. Analysis of resistance determinants in S. enterica strains (n = 33 revealed that the resistance to β-lactam antibiotics, trimethoprim-sulfamethoxazole, chloramphenicol, and tetracycline, was attributed to the presence of carb-like, dfrA1, floR, tetA gene, respectively. On the other hand, fluoroquinolone resistance was related to the presence of mutations in gyrA and parC genes. These findings improve the information about foodborne Salmonella in Saudi Arabia, alarming the emergence of multi-drug resistant S. enterica strains, and provide useful data about the resistance mechanisms.
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...
El-Tayeb, Mohamed A; Ibrahim, Abdelnasser S S; Al-Salamah, Ali A; Almaary, Khalid S; Elbadawi, Yahya B
Salmonella is recognized as a common foodborne pathogen, causing major health problems in Saudi Arabia. Herein, we report epidemiology, antimicrobial susceptibility and the genetic basis of resistance among S. enterica strains isolated in Saudi Arabia. Isolation of Salmonella spp. from clinical and environmental samples resulted in isolation of 33 strains identified as S. enterica based on their biochemical characteristics and 16S-rDNA sequences. S. enterica serovar Enteritidis showed highest prevalence (39.4%), followed by S. Paratyphi (21.2%), S. Typhimurium (15.2%), S. Typhi and S. Arizona (12.1%), respectively. Most isolates were resistant to 1st and 2nd generation cephalosporin; and aminoglycosides. Moreover, several S. enterica isolates exhibited resistance to the first-line antibiotics used for Salmonellosis treatment including ampicillin, trimethoprim-sulfamethoxazole and chloramphenicol. In addition, the results revealed the emergence of two S. enterica isolates showing resistance to third-generation cephalosporin. Analysis of resistance determinants in S. enterica strains (n=33) revealed that the resistance to β-lactam antibiotics, trimethoprim-sulfamethoxazole, chloramphenicol, and tetracycline, was attributed to the presence of carb-like, dfrA1, floR, tetA gene, respectively. On the other hand, fluoroquinolone resistance was related to the presence of mutations in gyrA and parC genes. These findings improve the information about foodborne Salmonella in Saudi Arabia, alarming the emergence of multi-drug resistant S. enterica strains, and provide useful data about the resistance mechanisms. Copyright © 2017. Published by Elsevier Editora Ltda.
Saltykova, Assia; Wuyts, Véronique; Mattheus, Wesley; Bertrand, Sophie; Roosens, Nancy H. C.; Marchal, Kathleen
Whole genome sequencing represents a promising new technology for subtyping of bacterial pathogens. Besides the technological advances which have pushed the approach forward, the last years have been marked by considerable evolution of the whole genome sequencing data analysis methods. Prior to application of the technology as a routine epidemiological typing tool, however, reliable and efficient data analysis strategies need to be identified among the wide variety of the emerged methodologies. In this work, we have compared three existing SNP-based subtyping workflows using a benchmark dataset of 32 Salmonella enterica subsp. enterica serovar Typhimurium and serovar 1,4,,12:i:- isolates including five isolates from a confirmed outbreak and three isolates obtained from the same patient at different time points. The analysis was carried out using the original (high-coverage) and a down-sampled (low-coverage) datasets and two different reference genomes. All three tested workflows, namely CSI Phylogeny-based workflow, CFSAN-based workflow and PHEnix-based workflow, were able to correctly group the confirmed outbreak isolates and isolates from the same patient with all combinations of reference genomes and datasets. However, the workflows differed strongly with respect to the SNP distances between isolates and sensitivity towards sequencing coverage, which could be linked to the specific data analysis strategies used therein. To demonstrate the effect of particular data analysis steps, several modifications of the existing workflows were also tested. This allowed us to propose data analysis schemes most suitable for routine SNP-based subtyping applied to S. Typhimurium and S. 1,4,,12:i:-. Results presented in this study illustrate the importance of using correct data analysis strategies and to define benchmark and fine-tune parameters applied within routine data analysis pipelines to obtain optimal results. PMID:29408896
Chadfield, M. S.; Christensen, J. P.; Madsen, Mogens
strain. The remaining putative 6, 7:-:- strain could not be identified and was genuinely non-motile. Diagnostic procedures performed initially were thus insufficient to differentiate between the different levels of motility and also resulted in mis-serotyping. As similar observations were made with two...
Charles L. Hofacre
Full Text Available Accurate and fast detection of harmful Salmonella is a major concern of food safety. Common Salmonella serotypes responsible for human associated foodborne outbreaks are S. Enteritidis, S. Hadar, S. Heidelberg, and S. Typhimurium are also commonly isolated from poultry. Serology is commonly used to monitor disease in poultry, therefore application of Salmonella serotype-specific test will have added value in Salmonella surveillance or monitoring vaccine efficacy. Recombinant flagellins were purified to be used as antigens in an ELISA. In this study, an ELISA was developed for the serological detection of S. Enteritidis. Once optimized, 500 ng of purified recombinant S. Enteritidis flagellin and a 1:64 dilution were determined to be optimal for testing sera. A negative baseline cutoff was calculated to be an optical density (OD of 0.35. All sera from birds with history of S. Enteritidis exposure tested positive and all sera from chickens with no exposure tested negative to this Salmonella serotype. Current ELISA for serological detection of Salmonella suffers from cross reactivity inherent in lipopolysaccharide (LPS or whole cell antigen based serological tests. This new ELISA eliminates common cross reactivity by focusing specifically on the flagellins of the Salmonella serotypes common in poultry and associated with foodborne outbreaks.
Braun, Sascha D.; Ziegler, Albrecht; Methner, Ulrich; Slickers, Peter; Keiling, Silke; Monecke, Stefan; Ehricht, Ralf
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
Santillana Farakos, Sofia; Hicks, J W; Frye, J G; Frank, J F
Salmonella enterica is not able to grow at water activity (aw) levels below 0.94, but it can survive in low-aw foods for long periods of time. Temperature, aw, substrate, and serotype affect its persistence. The aim of this study was to evaluate the influence of temperature and aw on the relative persistence among four serotypes of Salmonella enterica in low-aw whey protein powder. Whey protein powder was equilibrated to aws 0.18 ± 0.02 and 0.54 ± 0.03, inoculated with a cocktail of Salmonella serovars (Agona, Tennessee, Montevideo, and Typhimurium), vacuum sealed, and stored at 36°C for 6 months and at 70°C for 48 h. Presumptive Salmonella colonies (30 to 32) were randomly picked from each plate at the end of each survival study. PCR multiplex serotyping was used to identify the isolates. A multinomial mixed logistic model with Salmonella Tennessee as a reference was used to test for significant differences in frequency distribution of the surviving serotypes. Salmonella Tennessee and Salmonella Agona were the most prevalent surviving serotypes, followed in decreasing order by Salmonella Montevideo and Salmonella Typhimurium. Statistical analysis indicated that temperature (P = 0.003) and aw (P = 0.012) influenced the relative prevalence of the Salmonella serotypes. If other environmental conditions are equal, Salmonella Tennessee is better able to survive than Salmonella Montevideo and Salmonella Typhimurium at higher temperatures and higher aw levels in low-aw whey protein powder held at 36 and 70°C. The relative prevalence of Salmonella Agona to Salmonella Tennessee did not change with increasing temperature (P = 0.211) or aw (P = 0.453). These results should be considered in risk assessment and when developing predictive models for survival of Salmonella in low-aw foods.
Bolton, Lance F.; Kelley, Lynda C.; Lee, Margie D.; Fedorka-Cray, Paula J.; Maurer, John J.
Salmonella enterica serotype typhimurium (S. typhimurium) DT104 (DT104) first emerged as a major pathogen in Europe and is characterized by its pentadrug-resistant pattern. It has also been associated with outbreaks in the United States. The organism typically carries resistance to ampicillin, chloramphenicol, streptomycin, sulfonamides, and tetracycline. The mechanism of chloramphenicol resistance in DT104 was determined by producing antibiotic-resistant Escherichia coli host strain clones from DT104 DNA. DNA from chloramphenicol-resistant clones was sequenced, and probes specific for the genes floS. typhimurium (floSt), int, invA, and spvC were produced for colony blot hybridizations. One hundred nine Salmonella isolates, including 44 multidrug-resistant DT104 isolates, were tested to evaluate the specificities of the probes. The gene floSt, reported in this study, confers chloramphenicol and florfenicol resistance on S. typhimurium DT104. Florfenicol resistance is unique to S. typhimurium DT104 and multidrug-resistant S. typhimurium isolates with the same drug resistance profile among all isolates evaluated. Of 44 DT104 isolates tested, 98% were detected based on phenotypic florfenicol resistance and 100% had the floSt-positive genotype. Resistances to florfenicol and chloramphenicol are conferred by the gene floSt, described in this paper. Presumptive identification of S. typhimurium DT104 can be made rapidly based on the presence of the floSt gene or its resulting phenotype. PMID:10203484
Gu, W; Vieira, A R; Hoekstra, R M; Griffin, P M; Cole, D
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.
Brown, D. J.; Baggesen, Dorte Lau; Platt, D. J.
The plasmid pOG670, a 54 kb, conjugative plasmid that specifies resistance to ampicillin and kanamycin and belonging to the incompatibility group X (IncX), was transferred into 10 isolates of Salmonella enterica serotype Enteritidis belonging to 10 different phage types (PT1, 2, 3, 4, 8, 9, 9b, 10......, 11 and 13). Acquisition of the plasmid by these strains did not result in the loss of any resident plasmids but resulted in phage type conversion in 8 of the 10 strains (PT1, 2, 4, 8, 9, 9b, 10 and 11). The observed changes in phage type were found to result from the loss of sensitivity to 3...... of the 10 typing phages used (phages 3, 5 and 7). Where the conversion resulted in a change to a defined phage type, both the new and original PTs belonged to the same, previously described, evolutionary lines. Enteritidis PTs 1, 4 and 8, commonly associated with poultry world-wide, were converted to PTs 21...
Onyango, David M; Ndeda, Violet Mmbone; Wandili, Sarah A; Wawire, Sifuna A; Ochieng, Phillip
Salmonella enterica subspecies enterica serovar Choleraesuis is a host-adapted, facultative, intracellular pathogen that causes swine paratyphoid. Its antimicrobial resistance presents a challenge to feed manufacturing industries. However, stopping antibiotics in animal feed would have economic implications for the industry. Conventional microbial methods for isolation and identification of S. Choleraesuis were employed. The isolates were subjected to screening against 17 antimicrobial agents and genotyping of resistance markers by PCR. The data were then analyzed and presented in percentages. Phenotypically, 43 out of 95 isolates showed multidrug resistance. Among the 17 antibiotics tested, resistance was observed as follows: sulphonamides (45.2%), nalidixic acid (44.25%), tetracycline (42%), ampicillin (36.8%), erythromycin (34.7%), carbenicillin (31.5%), chrolamphenical (28.4%), gentamicin (27.3%), kanamycin (24.2%), spectinomycin (21%), sulfamethoxazole-trimethoprim (16.8%), streptomycin (12.6%), cephalothion (8.4%), ofloxacin (5.2%), ciprofloxacin (4.2%), and norfloxacin (4.2%). Fifty-two isolates were susceptible to the antimicrobial agents tested. A total of 3.1% of the isolates had the integron gene pattern combination of dfrA2-aadA2 (2100 bp), dfrA12 (2100 bp); 4.2% had dfrA12-aadA2-sulI (2100 bp); 2.1% had dfrA12-aadA2 (2100 bp); and 1% had dfrA2-aadA2-sulI (2100 bp), oxa1-aadA2 (1500 bp), dfrA12-aadA2-sulI, and blaPSE (2100 bp). The isolated S. Choleraesuis were resistant to more than 10% of the antimicrobial agents used in this study. Appropriate surveillance is warranted to gain more information about the epidemiology, as stopping antibiotics in animal feed would have economic implications for the industry.
"Multiplex PCR" identification of the atypical and monophasic Salmonella enterica subsp. enterica serotype 1,4,,12:i:- in São Paulo State, Brazil: frequency and antibiotic resistance patterns Identificação por "Multiplex PCR" do sorotipo monofásico e atípico Salmonella enterica subsp. enterica sorotipo 1,4,,12:i:-, no Estado de São Paulo, Brasil: freqüência e resistência antimicrobiana
Ana T. Tavechio
Full Text Available Salmonella spp. are the etiologic agents of salmonellosis, a worldwide spread zoonoses causing foodborne outbreaks and clinical diseases. By serological identification, Salmonella enterica subsp. enterica serotype 1,4,,12:i:- accounted for 8.8% of human and 1.6% of nonhuman Salmonella strains isolated in São Paulo State, during 1991-2000. A total of 28.6% of them amplified a fragment corresponding to H:1,2 (flagellar phase two through PCR analysis and were further assigned as S. Typhimurium. Antimicrobial resistance was detected in 36.3% of the 369 PCR-negative strains tested, including the multiresistance to ampicillin, chloramphenicol, sulfonamides, tetracycline, and streptomycin.Salmonella spp. é o agente etiológico da salmonelose, zoonose mundialmente distribuída e responsável por surtos de doenças transmitidas por alimentos e doenças clínicas. Sorologicamente, Salmonella enterica subsp. enterica sorotipo 1,4,,12:i:- correspondeu a 8,8% e 1,6% das cepas de Salmonella de origem humana e não-humana, respectivamente, isoladas no Estado de São Paulo, no decênio 1991-2000. Aproximadamente 28,6% destas cepas amplificaram o fragmento correspondente a H:1,2 (fase flagelar dois em testes de PCR e foram, então, identificadas como S. Typhimurium. Das 369 cepas negativas em PCR, 36,3% apresentou resistência antimicrobiana, incluindo multirresistência a ampicilina, cloranfenicol, sulfonamidas, tetraciclina e estreptomicina.
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.
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.
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
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,,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
Yao, Kuan; Muruvanda, Tim; Roberts, Richard J.; Payne, Justin; Allard, Marc W.; Hoffmann, Maria
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 S...
Outbreak epidemiologically linked with a composite product of beef, mechanically separated chicken and textured vegetable protein contaminated with multiple serotypes of Salmonella enterica including multidrug-resistant Infantis, California 2016.
Hutchinson, J A; Wheeler, C; Mohle-Boetani, J C
A salmonellosis outbreak occurred at a California prison in April and May 2016. In a cohort study of 371 inmates, persons who consumed dishes from the prison kitchen made from ground meat had a higher attack rate (15%) than those who did not (4%) (risk ratio 3.4, 95% CI 1.1-10.6). The ground meat product was composed exclusively of beef, mechanically separated chicken (MSC) and textured vegetable protein; eight of eight lots of the product collected from the prison and processing facility were contaminated with Salmonella enterica of eight serotypes and 17 distinct PFGE patterns, including multidrug-resistant S. Infantis. Either the MSC or the beef could have been the source of the particular strains of S. enterica isolated from patients or the product. The microbiological evidence is most consistent with MSC as the source of the high levels of S. enterica in the epidemiologically linked meat product. Our findings contribute to the growing body of evidence about the hazard posed by the use of products containing raw mechanically separated poultry in kitchens in institutions.
Full Text Available 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.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.This is the first analysis of cellular immune responses to purified S. Typhi antigens in patients with typhoid fever. These results indicate
Full Text Available Abstract Background Salmonella enterica serotype Typhimurium produces surface-associated fimbriae that facilitate adherence of the bacteria to a variety of cells and tissues. Type 1 fimbriae with binding specificity to mannose residues are the most commonly found fimbrial type. In vitro, static-broth culture favors the growth of S. Typhimurium with type 1 fimbriae, whereas non-type 1 fimbriate bacteria are obtained by culture on solid-agar media. Previous studies demonstrated that the phenotypic expression of type 1 fimbriae is the result of the interaction and cooperation of the regulatory genes fimZ, fimY, fimW, and fimU within the fim gene cluster. Genome sequencing revealed a novel gene, stm0551, located between fimY and fimW that encodes an 11.4-kDa putative phosphodiesterase specific for the bacterial second messenger cyclic-diguanylate monophosphate (c-di-GMP. The role of stm0551 in the regulation of type 1 fimbriae in S. Typhimurium remains unclear. Results A stm0551-deleted stain constructed by allelic exchange constitutively produced type 1 fimbriae in both static-broth and solid-agar medium conditions. Quantative RT-PCR revealed that expression of the fimbrial major subunit gene, fimA, and one of the regulatory genes, fimZ, were comparably increased in the stm0551-deleted strain compared with those of the parental strain when grown on the solid-agar medium, a condition that normally inhibits expression of type 1 fimbriae. Following transformation with a plasmid possessing the coding sequence of stm0551, expression of fimA and fimZ decreased in the stm0551 mutant strain in both culture conditions, whereas transformation with the control vector pACYC184 relieved this repression. A purified STM0551 protein exhibited a phosphodiesterase activity in vitro while a point mutation in the putative EAL domain, substituting glutamic acid (E with alanine (A, of STM0551 or a FimY protein abolished this activity. Conclusions The finding that the
Full Text Available Salmonella is widespread in nature and can be found in all links of the poultry production chain. Due to its high impact on meat processing, techniques for the rapid detection and reproducible characterization of Salmonella serotypes in foods are needed. The present study investigated the potential of molecular profiling to identify and differentiate 15 Salmonella serotypes isolated from the poultry production chain, based on 5 primers by random amplified polymorphic DNA (RAPD, enterobacterial repetitive intergenic consensus (ERIC-PCR, amplification of rDNA internal spacer analysis (RISA, and amplified ribosomal DNA restriction analysis (ARDRA of 16S-23S rRNA internal spacer region (ISR cleaved with Alu I and Hha I restriction enzymes. Three isolates of each serotype were analyzed for the identification of similar and different profiles. Dendrograms were constructed from molecular profiles using the UPGMA method (unweighted pair-group method for the arithmetic averages and the software program WinBoot. The present study indicates the usefulness of RISA and ARDRA of the 16S-23S rRNA intergenic spacer region (ISR for systematic, epidemiological, and diagnostic purposes. Since these techniques can be used for the differentiation of serotypes, they are highly promising for the characterization of Salmonella serotypes and intra-serotypes. Data indicate that these techniques may be used to produce more consistent, reliable, and reproducible results in the identification and epidemiological study (traceability of Salmonella in the poultry industry.
Salmonella enterica Ituri is an uncommon serotype associated with poultry disease. One of the serotype isolated from a poultry disease in Nigeria was characterized by serotyping and screening for the presence of Salmonella genomic island 1(SGI1) as a possible factor responsible for its involvement in a poultry disease ...
Kamble, Nitin M; Lee, John Hwa
Natural infections of chickens with Salmonella enterica subsp. enterica serovar Senftenberg (S. Senftenberg) are characterized by low-level intestinal invasiveness and insignificant production of antibodies. In this study, we investigated the potential effects of lon and cpxR gene deletions on the invasiveness of S Senftenberg into the intestinal epithelium of chickens and its ability to induce an immune response, conferring protection against S Senftenberg infection. With the allelic exchange method, we developed JOL1596 (Δlon), JOL1571 (ΔcpxR), and JOL1587 (Δlon ΔcpxR) deletion mutants from wild-type S Senftenberg. Deletion of the lon gene from S Senftenberg produced increased frequency of elongated cells, with significantly greater amounts of exopolysaccharide (EPS) than in the cpxR-deleted strain and the wild-type strain. The in vivo intestinal loop invasion assay showed a significant increase in epithelial invasiveness for JOL1596 (Δlon) and JOL1587 (Δlon ΔcpxR), compared to JOL1571 (ΔcpxR) and the wild-type strain. Furthermore, the S Senftenberg wild-type and mutant strains were internalized at high levels inside activated abdominal macrophages from chicken. The in vivo inoculation of JOL1587 (Δlon ΔcpxR) into chickens led to colonization of the liver, spleen, and cecum for a short time. Chickens inoculated with JOL1587 (Δlon ΔcpxR) showed significant increases in humoral, mucosal, and cellular immune responses specific to S Senftenberg antigens. Postchallenge, compared to the control group, the JOL1587 (Δlon ΔcpxR)-inoculated chickens showed not only lower persistence but also faster clearance of wild-type S Senftenberg from the cecum. We conclude that the increased intestinal invasiveness and colonization of internal organs exhibited by JOL1587 (Δlon ΔcpxR) led to the establishment of immunogenicity and conferred protective efficacy against S Senftenberg infections in chickens. Copyright © 2016, American Society for Microbiology. All Rights
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
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.
Salmonella enteric subsp. enterica strains RM11060, serotype 6,8:d:-, and RM11065, serotype 6,8:-:e,n,z15, were isolated from environmental sampling in Central California in 2009. We report the complete genome sequences and annotation of these two strains. These genomic sequences are distinct and wi...
Influence of Saccharomyces cerevisiae fermentation products, SmartCare in milk replacer and Original XPC in calf starter, on the performance and health of preweaned Holstein calves challenged with Salmonella enterica serotype
This study was designed to investigate the effects of supplementing SmartCare™ (SC) in milk replacer and Original XPC™(XPC) in calf starter on performance and health of preweaned calves following an oral challenge with Salmonella enterica. The study was performed in two 35-d periods with 30 Holstein...
Wattiau, Pierre; Boland, Cécile; Bertrand, Sophie
For more than 80 years, subtyping of Salmonella enterica has been routinely performed by serotyping, a method in which surface antigens are identified based on agglutination reactions with specific antibodies. The serotyping scheme, which is continuously updated as new serovars are discovered, has generated over time a data set of the utmost significance, allowing long-term epidemiological surveillance of Salmonella in the food chain and in public health control. Conceptually, serotyping provides no information regarding the phyletic relationships inside the different Salmonella enterica subspecies. In epidemiological investigations, identification and tracking of salmonellosis outbreaks require the use of methods that can fingerprint the causative strains at a taxonomic level far more specific than the one achieved by serotyping. During the last 2 decades, alternative methods that could successfully identify the serovar of a given strain by probing its DNA have emerged, and molecular biology-based methods have been made available to address phylogeny and fingerprinting issues. At the same time, accredited diagnostics have become increasingly generalized, imposing stringent methodological requirements in terms of traceability and measurability. In these new contexts, the hand-crafted character of classical serotyping is being challenged, although it is widely accepted that classification into serovars should be maintained. This review summarizes and discusses modern typing methods, with a particular focus on those having potential as alternatives for classical serotyping or for subtyping Salmonella strains at a deeper level. PMID:21856826
Salmonella enterica Give is one of the serotypes that have been incriminated in Salmonella infections; sometimes associated with hospitalization and mortalities in humans and animals in some parts of the world. In this work, we characterized one Salmonella Give isolated from cloaca swab of an Agama agama lizard ...
Raufu, Ibrahim A; Odetokun, Ismail A; Oladunni, Fatai S; Adam, Mohammed; Kolapo, Ubaidat T; Akorede, Ganiu J; Ghali, Ibraheem M; Ameh, James A; Ambali, Abdulganiyu
This study aimed at determining the serotypes, antimicrobial profiles, and public health importance of Salmonella strains from camels slaughtered at Maiduguri central abattoir, Nigeria. Two hundred samples were obtained from camel comprising of intestines, feces, liver, and spleen (n=50 each). Non-lactose fermenting dark center Salmonella colonies were identified using standard biochemical techniques, serotyped and subjected to antimicrobial susceptibility test using minimum inhibition concentration method. Out of the 200 samples collected, 17 were Salmonella positive (spleen=7, intestine=6, feces=3, and liver=1) with a prevalence of 8.5%. Five serotypes comprising Salmonella Eko, 7 (3.5%), Salmonella Uganda, 4 (2.0%), Salmonella Amager, 2 (1.0%), Salmonella Westhampton, 2 (1.0%), and Salmonella Give, 2 (1.0%) were incriminated. Majority of the serotypes were sensitive to the antimicrobials, but one Salmonella Amager exhibited resistance to streptomycin, and one each of Salmonella Uganda and Salmonella Eko were resistant to sulfamethoxazole. This study revealed the prevalence and the antibiotic resistance profile of newly emerging Salmonella from camels in the northeast of Nigeria, which can serve as a means for the transmission of Salmonella to human. Therefore, there is a need for the establishment of national Salmonella surveillance and control programs.
Soyer, Yeşim; Orsi, Renato H; Rodriguez-Rivera, Lorraine D; Sun, Qi; Wiedmann, Martin
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. 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] positive selection (FDR 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. 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) branch specific positive selection contributes to the evolution of host restricted Salmonella serotypes.
Jacobsen, Annika; Hendriksen, Rene S.; Aarestrup, Frank Møller
Salmonella enterica is divided into four subspecies containing a large number of different serovars, several of which are important zoonotic pathogens and some show a high degree of host specificity or host preference. We compare 45 sequenced S. enterica genomes that are publicly available (22......, and the core and pan-genome of Salmonella were estimated to be around 2,800 and 10,000 gene families, respectively. The constructed pan-genomic dendrograms suggest that gene content is often, but not uniformly correlated to serotype. Any given Salmonella strain has a large stable core, whilst...... there is an abundance of accessory genes, including the Salmonella pathogenicity islands (SPIs), transposable elements, phages, and plasmid DNA. We visualize conservation in the genomes in relation to chromosomal location and DNA structural features and find that variation in gene content is localized in a selection...
Full Text Available Abstract 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] 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 branch specific positive selection contributes to the evolution of host restricted Salmonella serotypes.
Zhou, Xiujuan; Zhang, Lida; Shi, Chunlei; Fratamico, Pina M; Liu, Bin; Paoli, George C; Dan, Xianlong; Zhuang, Xiaofei; Cui, Yan; Wang, Dapeng; Shi, Xianming
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 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.
Bakhshi, Bita; Ghafari, Mohsen; Pourshafie, Mohammad R; Zarbakhsh, Behnaz; Katouli, Mohammad; Rahbar, Mohammad; Hajia, Masoud; Hosseini-Aliabad, Neda; Boustanshenas, Mina
The epidemiology of salmonellosis is complex because of the diversity and different serotypes of Salmonella enterica (S. enterica) that occur in different reservoirs and geographic incidences. To determine the genotype distribution and resistance-gene content of 2 classes of integron among S. enterica isolates. Thirty-six S. enterica species were isolated and tested for their serological distribution and the resistance-gene contents of 2 classes of integron, as well as for their genetic diversity, using the pulsed-field gel electrophoresis (PFGE) genotyping method. Serogroups E (36.1%) and D (30.5%) were dominant among the isolates. All of the isolates in serogroup D belonged to the serovar enteritidis. The aadA1 gene was found within all resistance-gene cassettes. We observed 4 common and 26 single pulsotypes among the isolates, which indicated a high degree of genetic diversity among the isolates. Using the PulseNet International standard protocol, it was found that these isolates were different from those reported previously in Iran. The presence of a few common and new pulsotypes among the isolates suggests the emergence and spread of new clones of S. enterica in Iran. Copyright© by the American Society for Clinical Pathology (ASCP).
Baggesen, Dorte Lau; Wegener, Henrik Caspar; Bager, Flemming
enterica were isolated from 832 pigs (6.2%). The predominant serotype was S. Typhimurium, comprising 536 (64.4%) of the isolates. Four hundred and forty-eight isolates of S. Typhimurium were examined by phage typing, resulting in detection of 17 different phage types (definitive types, DT) with DT12 being...
Carlson, James C; Hyatt, Doreene R; Bentler, Kevin; Mangan, Anna M; Russell, Michael; Piaggio, Antoinette J; Linz, George M
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. Published by Elsevier B.V.
Dieckmann, Ralf; Malorny, Burkhard
Currently, 2,610 different Salmonella serovars have been described according to the White-Kauffmann-Le Minor scheme. They are routinely differentiated by serotyping, which is based on the antigenic variability at lipopolysaccharide moieties (O antigens), flagellar proteins (H1 and H2 antigens), and capsular polysaccharides (Vi antigens). The aim of this study was to evaluate the potential of matrix-assisted laser desorption ionization–time of flight (MALDI-TOF) mass spectrometry for rapid screening and identification of epidemiologically important Salmonella enterica subsp. enterica serovars based on specific sets of serovar-identifying biomarker ions. By analyzing 913 Salmonella enterica subsp. enterica strains representing 89 different serovars using MALDI-TOF mass spectrometry, several potentially serovar-identifying biomarker ions were selected. Based on a combination of genus-, species-, subspecies-, and serovar-identifying biomarker ions, a decision tree classification algorithm was derived for the rapid identification of the five most frequently isolated Salmonella enterica serovars, Enteritidis, Typhimurium/4,,12:i:-, Virchow, Infantis, and Hadar. Additionally, sets of potentially serovar-identifying biomarker ions were detected for other epidemiologically interesting serovars, such as Choleraesuis, Heidelberg, and Gallinarum. Furthermore, by using a bioinformatic approach, sequence variations corresponding to single or multiple amino acid exchanges in several biomarker proteins were tentatively assigned. The inclusivity and exclusivity of the specific sets of serovar-identifying biomarker ions for the top 5 serovars were almost 100%. This study shows that whole-cell MALDI-TOF mass spectrometry can be a rapid method for prescreening S. enterica subsp. enterica isolates to identify epidemiologically important serovars and to reduce sample numbers that have to be subsequently analyzed using conventional serotyping by slide agglutination techniques. PMID
Arnott, Alicia; Wang, Qinning; Bachmann, Nathan; Sadsad, Rosemarie; Biswas, Chayanika; Sotomayor, Cristina; Howard, Peter; Rockett, Rebecca; Wiklendt, Agnieszka; Iredell, Jon R; Sintchenko, Vitali
Multidrug- and colistin-resistant Salmonella enterica serotype 4,,12:i:- sequence type 34 is present in Europe and Asia. Using genomic surveillance, we determined that this sequence type is also endemic to Australia. Our findings highlight the public health benefits of genome sequencing-guided surveillance for monitoring the spread of multidrug-resistant mobile genes and isolates.
Arnott, Alicia; Wang, Qinning; Bachmann, Nathan; Sadsad, Rosemarie; Biswas, Chayanika; Sotomayor, Cristina; Howard, Peter; Rockett, Rebecca; Wiklendt, Agnieszka; Iredell, Jon R.
Multidrug- and colistin-resistant Salmonella enterica serotype 4,,12:i:- sequence type 34 is present in Europe and Asia. Using genomic surveillance, we determined that this sequence type is also endemic to Australia. Our findings highlight the public health benefits of genome sequencing–guided surveillance for monitoring the spread of multidrug-resistant mobile genes and isolates. PMID:29553318
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
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. Copyright © 2015 Japanese Society of Chemotherapy and The Japanese Association for Infectious Diseases. Published by Elsevier Ltd. All rights reserved.
Karns, Jeffrey S; Haley, Bradd J; Van Kessel, Jo Ann S
Molecular serotyping through the use of PCR is a simple and useful technique for characterizing isolates of Salmonella enterica subsp. enterica belonging to serogroups B, C1, C2, D1, and E1, which are the majority of the isolates associated with human disease outbreaks. However, many of the Salmonella strains currently isolated from dairy farms in the northeastern United States are serovar Cerro, a group K strain not detected by this assay. Primers from a well-known PCR assay for the identification of Salmonella were added to a commonly used serotyping assay so that strains, such as Salmonella Cerro, that do not produce bands in the original assay can be confirmed as belonging to S. enterica subsp. enterica. The modified assay frequently misidentified the serogroup of Salmonella Mbandaka isolates because of failure to amplify the wzxC1 amplicon. Therefore, the reverse primer for the wzxC1 target was modified based on in silico analysis to provide consistent classification of Salmonella Mbandaka as belonging to serogroup C1. These two modifications to the serogrouping PCR method enhance the utility of the method for characterizing Salmonella isolates.
Davis, A L; Curtis, P A; Conner, D E; McKee, S R; Kerth, L K
Salmonella enterica serotype Enteritidis has long been associated with eggs, and more recently, Salmonella enterica serotype Heidelberg has also become associated with eggs. This study was undertaken to determine whether Salmonella Enteritidis and Salmonella Heidelberg are effectively eliminated from eggs by various cooking methods. Seven cooking methods were chosen--hard and soft cooked, scrambled, over easy, sunny-side up, poached, and free poached--and a pan insert and the free-flowing method were used. Shell eggs, purchased from a grocery store, were inoculated with Salmonella and cooked. The cooked eggs were analyzed by USDA-approved methods for Salmonella recovery. Findings indicated that existing cooking methods for the hard-cooked, soft-cooked, and poaching methods were safe. However, the same was not true for the current sunny-side-up, over-easy, and scrambled egg cooking methods.
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
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. Published by Elsevier B.V.
Full Text Available Salmonella infections are the second leading cause of zoonotic bacterial foodborne illness. Main source of infection in human is contaminated food products. The aim of this study was sub typing isolates of Salmonella enterica obtained during our previous study by Pulsed Field Gel Electrophoresis (PFGE technique. All 46 Salmonella isolates were serotyped and then subjected to PFGE. Total isolates were analyzed by means of the molecular technique XbaI PFGE. In this study, PFGE and serotyping were used to subtype 46 Salmonella isolates belonging to 27different serovars and derived from human and different food origins. Among these isolates, S. Typhimurium was found to be the most predominant serovar. 40 PFGE patterns out of 46 isolates were obtained. The Discrimination Index obtained by serotyping (DI = 0.93 was lower than PFGE (DI = 0.99. Subtyping of Salmonella enterica is very important and shows that animal origin can be one of a reservoir that potentially could be transferred to human through the food chain. In addition, results of this study also revealed that this procedure is a golden standard for genotyping of such salmonella serotypes.
Drolet, Barbara S.; Reister-Hendricks, Lindsey M.; Podell, Brendan K.; Breitenbach, Jonathan E.; Mcvey, D.S.; Rijn, van Piet A.; Bowen, Richard A.
Bluetongue virus (BTV) is an orbivirus transmitted by biting midges (Culicoides spp.) that can result in moderate to high morbidity and mortality primarily in sheep and white-tailed deer. Although only 5 serotypes of BTV are considered endemic to the United States, as many as 11 incursive
Ren, X; Li, M; Xu, C; Cui, K; Feng, Z; Fu, Y; Zhang, J; Liao, M
A total of 1145 samples were collected from chicken breeder farms, hatcheries, broiler farms, a slaughterhouse and retail refrigerated chicken stores in an integrated broiler supply chain in Guangdong Province, China, in 2013. One-hundred and two Salmonella enterica strains were isolated and subjected to serotyping, antimicrobial susceptibility testing, virulence profile determination and molecular subtyping by pulsed field gel electrophoresis (PFGE). The contamination rates in samples from breeder farms, hatcheries, broiler farms, the slaughterhouse and retail stores were 1·46%, 4·31%, 7·00%, 62·86% and 54·67%, respectively. The isolated strains of S. enterica belonged to 10 serotypes; most of them were S. Weltevreden (46·08%, 47/102) and S. Agona (18·63%, 19/102). Isolates were frequently resistant to streptomycin (38·2%), tetracycline (36·3%), sulfisoxazole (35·3%) and gentamicin (34·3%); 31·4% of isolates were multidrug resistant. The isolates were screened for 10 virulence factors. The Salmonella pathogenicity island genes avrA, ssaQ, mgtC, siiD, and sopB and the fimbrial gene bcfC were present in 100% of the strains. PFGE genotyping of the 102 S. enterica isolates yielded 24 PFGE types at an 85% similarity threshold. The PFGE patterns show that the genotypes of S. enterica in the production chain are very diverse, but some strains have 100% similarity in different parts of the production chain, which indicates that some S. enterica persist throughout the broiler supply chain.
Kjeldsen, Marianne Kirstine; Torpdahl, Mia; Pedersen, Karl
Aims Salmonella enterica subsp. enterica causes salmonellosis in humans and animals. Serovar specific multiple-locus variable-number tandem repeat analysis (MLVA) is widely used for Salmonella surveillance; however, isolates have to be serotyped prior to MLVA typing and only the most common...... serovars can be typed. We developed a MLVA scheme for high discriminatory typing of Salmonella. Methods and Results Sixty-six unique VNTRs were investigated and the polymorphisms of seven promising VNTRs were evaluated with a panel 163 diverse isolates of 14 serotypes of significance for human health. Five......-related strains. Conclusions The technique showed a high discriminatory power within most serotypes comparable with or better than that of PFGE. Significance and impact of the Study This MLVA assay makes it possible to use a single typing method for Salmonella surveillance and outbreak investigations. This allows...
Antunes, P; Campos, J; Mourão, J; Ribeiro, T G; Novais, C; Peixe, L
Non-typhoidal Salmonella is an important burden, particularly in developing countries of the African region. We report for the first time in Angola, a sub-Saharan African country with commercial/travel relationships with Europe, an unexpectedly high occurrence of Salmonella (n = 12/63, 19%) from a high diversity of sources, particularly farm and wild animals. The detection of diverse serotypes (n = 12), involving putative new S. enterica subsp. salamae serotypes, is also of note, reinforcing the need for a comprehensive surveillance in Angola critical to identify animal/food/environmental sources of salmonellosis with impact on animal health, local people, tourists and exported products.
Lamas, A; Miranda, J M; Vázquez, B; Cepeda, A; Franco, C M
Salmonella enterica subsp. enterica is one of the main food-borne pathogens. This microorganism combines an aerobic life outside the host with an anaerobic life within the host. One of the main concerns related to S. enterica is biofilm formation and cellulose production. In this study, biofilm formation, morphotype, cellulose production and transcription of biofilm and quorum sensing-related genes of 11 S. enterica strains were tested under three different conditions: aerobiosis, microaerobiosis, and anaerobiosis. The results showed an influence of oxygen levels on biofilm production. Biofilm formation was significantly higher (Penterica strains tested. This gene expression levels were less reduced in S. Typhimurium and S. Enteritidis compared to the tested serotypes. There was a relationship between the expression of biofilm and quorum sensing-related genes. Thus, the results from this study indicate that biofilm formation and cellulose production are highly influenced by atmospheric conditions. This must be taken into account as contamination with these bacteria can occur during food processing under vacuum or modified atmospheres. Copyright © 2016 Elsevier B.V. All rights reserved.
Nurhonen, Markku; Auranen, Kari
Pneumococcal conjugate vaccination has proved highly effective in eliminating vaccine-type pneumococcal carriage and disease. However, the potential adverse effects of serotype replacement remain a major concern when implementing routine childhood pneumococcal conjugate vaccination programmes. Applying a concise predictive model, we present a ready-to-use quantitative tool to investigate the implications of serotype replacement on the net effectiveness of vaccination against invasive pneumococcal disease (IPD) and to guide in the selection of optimal vaccine serotype compositions. We utilise pre-vaccination data on pneumococcal carriage and IPD and assume partial or complete elimination of vaccine-type carriage, its replacement by non-vaccine-type carriage, and stable case-to-carrier ratios (probability of IPD per carriage episode). The model predicts that the post-vaccination IPD incidences in Finland for currently available vaccine serotype compositions can eventually decrease among the target age group of children replacement through herd effects, the decrease among the older population is predicted to be much less (20-40%). We introduce a sequential algorithm for the search of optimal serotype compositions and assess the robustness of inferences to uncertainties in data and assumptions about carriage and IPD. The optimal serotype composition depends on the age group of interest and some serotypes may be highly beneficial vaccine types in one age category (e.g. 6B in children), while being disadvantageous in another. The net effectiveness will be improved only if the added serotype has a higher case-to-carrier ratio than the average case-to-carrier ratio of the current non-vaccine types and the degree of improvement in effectiveness depends on the carriage incidence of the serotype. The serotype compositions of currently available pneumococcal vaccines are not optimal and the effectiveness of vaccination in the population at large could be improved by including
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.
Marzel, Alex; Desai, Prerak T; Nissan, Israel; Schorr, Yosef Ilan; Suez, Jotham; Valinsky, Lea; Reisfeld, Abraham; Agmon, Vered; Guard, Jean; McClelland, Michael; Rahav, Galia; Gal-Mor, Ohad
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. Copyright © 2014, American Society for Microbiology. All Rights Reserved.
Full Text Available Background and Objectives: Chloramphenicol was considered the anti-microbial gold standard for typhoid treatment but, following the increasing worldwide frequency of antibiotic resistance, ciprofloxacin has been the mainstay of therapy since 1980. Recent studies have shown a shifting of susceptibility to conventional drugs like chloramphenicol, ampicillin and cotrimoxazole. The primary objective of the study was to evaluate the in vitro activity of chloramphenicol and other first-line drugs in comparison with cephalosporins and quinolones. Materials and Methods: Fifty isolates of Salmonella obtained from blood culture were subjected to serotyping at the Central Research Institute, Kasauli. Phage typing and biotyping was performed at the National Phage Typing Centre, New Delhi. Antibiotic sensitivity testing was carried out for 10 drugs by the Kirby-Bauer disc diffusion method and minimum inhibitory concentration by broth microdilution for nalidixic acid, chloramphenicol, ciprofloxacin, ceftriaxone, cefixime and ofloxacin. Multi-drug-resistant (MDR strains were checked for plasmid. Results: In the present study, 70 and 30% of the isolates were Salmonella enterica serovar typhi and paratyphi A, respectively. They were highly sensitive to chloramphenicol (86%, ampicillin (84% and cotrimoxazole (88%. Highest sensitivity was seen for cephalosporins, followed by quinolones. Seventeen/21 (81% and 100% of the Salmonella enterica serovar typhi strains belonged to E1 phage type and biotype 1, respectively. Antibiogram showed 2% of the strains to be sensitive to all the drugs tested and 12% were MDR and showed the presence of plasmids. Conclusion: The study indicates reemergence of chloramphenicol-susceptible Salmonella enterica serovar typhi and paratyphi A isolates, a significant decline in MDR strains and high resistance to nalidixic acid. E1 phage type and biotype 1 are found to be most prevalent in Chennai, India.
Foss, Dennis L; Agin, Tonia S; Bade, Donald; Dearwester, Don A; Jolie, Rika; Keich, Robin L; Lohse, Robert M; Reed, Maryke; Rosey, Everett L; Schneider, Patricia A; Taylor, Lucas P; Willy, Michael S
Pre-harvest reduction of Salmonella carriage by swine would benefit both animal health and food quality. While vaccination is an attractive pre-harvest intervention to reduce Salmonella levels in swine, the large number of potential Salmonella enterica serovars found in swine makes it critical that vaccines provide broad serotype efficacy. In order to directly compare the relative efficacy of Salmonella vaccines against serogroup-matched and serogroup-unmatched Salmonella, we vaccinated pigs with two commercially available Salmonella vaccines (either serogroup B or serogroup C1) and challenged with serovar-matched, serogroup-matched or serogroup-unmatched challenge strains. We found that while serogroup-matched vaccines provided relatively better efficacy than unmatched vaccines, serotype-unmatched vaccines also provided significant reduction of Salmonella carriage and shed. In addition, by measuring serogroup specific cell mediated (IFN-γ ELISPOT) and humoral (anti-LPS ELISA) immunity, we found that this serogroup specific efficacy correlates primarily with humoral immunity, while cell mediated immunity was mostly non-serogroup specific. While the practical relevance to pork quality of this serogroup-specific efficacy remains to be demonstrated, the large predominance of serogroup B Salmonella in swine suggests that a serogroup B Salmonella vaccine for swine would be of value to pre-harvest food safety interventions in swine. Copyright © 2013 Elsevier B.V. All rights reserved.
Valenzuela, J R; Sethi, A K; Aulik, N A; Poulsen, K P
Salmonellosis on the dairy continues to have a significant effect on animal health and productivity and in the United States. Additionally, Salmonella enterica ssp. enterica causes an estimated 1.2 million cases of human illness annually. Contributing to the morbidity and mortality in both human and domestic animal species is emergence of antimicrobial resistance by Salmonella species and increased incidence of multidrug-resistant isolates. This study describes serotype distribution and the antimicrobial resistance patterns for various Salmonella serotypes isolated from bovine samples submitted to the Wisconsin Veterinary Diagnostic Laboratory (WVDL) over the past 10 yr. Salmonella serotyping and antimicrobial susceptibility testing data were obtained from the laboratory information management system at WVDL. Data from accessions were limited to bovine samples submitted to the WVDL between January 2006 and June 2015 and those that had both a definitive serotype and complete results for antimicrobial susceptibility testing. A total of 4,976 isolates were identified. Salmonella enterica ser. Dublin was the most prevalent serotype identified among bovine samples submitted to the WVDL, accounting for a total of 1,153 isolates (23% of total isolates) over the study period. Along with Dublin, Salmonella enterica ser. Cerro (795, 16%), Newport (720, 14%), Montevideo (421, 8%), Kentucky (419, 8%), and Typhimurium (202, 4%) comprised the top 6 most commonly isolated serotypes during that time. Overall, resistance of bovine Salmonella isolates in the study population remained stable, although decreases in resistance were noted for gentamicin, neomycin, and trimethoprim sulfamethoxazole during the study period. All isolates remained susceptible to enrofloxacin. These data show that antimicrobial susceptibility for bovine Salmonella has changed in the population served by WVDL in the past 10 yr. This information is important for understanding Salmonella disease ecology in
Ruiz-Pina, Hugo Antonio; Puc-Franco, Miguel Angel; Flores-Abuxapqui, Javier; Vado-Solis, Ignacio; Cardenas-Marrufo, María Fidelia
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.
Abraham, Sam; Groves, Mitchell D; Trott, Darren J; Chapman, Toni A; Turner, Bernadette; Hornitzky, Michael; Jordan, David
Salmonella enterica is a zoonotic pathogen causing a variety of diseases in humans and animals. Many countries are reporting an increase in the prevalence of multidrug-resistant (MDR) S. enterica in food animals. The aim of this study was to determine whether S. enterica isolated from livestock in New South Wales, Australia, have similar resistance traits to those reported internationally. Salmonella enterica (n=165) from clinical infections in food animals between 2007 and 2011 were serotyped and tested for susceptibility to 18 antimicrobials. Also, 22 antimicrobial resistance genes (ARGs), 3 integrons and 18 plasmid replicon types were screened for using PCR. Most isolates (66.1%) remained susceptible to all antimicrobials; 8.5% of the isolates were resistant to four or more antimicrobials. Antimicrobials with the highest prevalence of resistance were sulfafurazole (28.5%), ampicillin (17.0%), tetracycline (15.8%) and trimethoprim (8.5%). There was no resistance to fluoroquinolones or third-generation cephalosporins. The most common ARGs were blaTEM (15.2%), sul2 (10.3%), tetB (9.1%), tetA (5.5%), aphA1 (4.8%) and dhfrV (4.8%). Class 1 integrons (7.9%) and IncFIIA (69.7%) were the most commonly detected integron and plasmid replicon types, respectively. Class 1 integrons were positively associated with MDR phenotypes and ARG carriage (P≤0.001). Internationally prominent MDR serovars associated with severe disease in humans (e.g. AmpC-positive Salmonella Newport) were not detected. Overall, the comparatively favourable resistance status of S. enterica in Australian livestock represents minimal public health risk associated with MDR strains and supports a conservative approach to the registration of antimicrobial drug classes in food-producing animals. Crown Copyright © 2013. Published by Elsevier B.V. All rights reserved.
Nurhonen, Markku; Auranen, Kari
Pneumococcal conjugate vaccination has proved highly effective in eliminating vaccine-type pneumococcal carriage and disease. However, the potential adverse effects of serotype replacement remain a major concern when implementing routine childhood pneumococcal conjugate vaccination programmes. Applying a concise predictive model, we present a ready-to-use quantitative tool to investigate the implications of serotype replacement on the net effectiveness of vaccination against invasive pneumococcal disease (IPD) and to guide in the selection of optimal vaccine serotype compositions. We utilise pre-vaccination data on pneumococcal carriage and IPD and assume partial or complete elimination of vaccine-type carriage, its replacement by non-vaccine-type carriage, and stable case-to-carrier ratios (probability of IPD per carriage episode). The model predicts that the post-vaccination IPD incidences in Finland for currently available vaccine serotype compositions can eventually decrease among the target age group of children vaccine types in one age category (e.g. 6B in children), while being disadvantageous in another. The net effectiveness will be improved only if the added serotype has a higher case-to-carrier ratio than the average case-to-carrier ratio of the current non-vaccine types and the degree of improvement in effectiveness depends on the carriage incidence of the serotype. The serotype compositions of currently available pneumococcal vaccines are not optimal and the effectiveness of vaccination in the population at large could be improved by including new serotypes in the vaccine (e.g. 22 and 9N). PMID:24550722
Full Text Available Background: Salmonella spp. are widespread zoonotic pathogens with economic importance for both humans and animals. They are categorized as the natural flora of the gastrointestinal tract of many reptiles. Human salmonellosis acquired from contact with reptiles is a well-recognized medical problem. Objective: The frequency of Salmonella contamination in Caspian pond turtles was surveyed to evaluate the danger of exposure or disease risk for humans as these species are abundant around the villages in Golestan and Mazandaran provinces. Materials and Methods: One hundred fifty fecal samples from Caspian pond turtles were tested by standard bacteriological methods and positive samples were serotyped. Antimicrobial susceptibility tests on isolated Salmonella strains were also performed. Results: Out of 150 samples, 54 turtles were detected to be contaminated with Salmonella. Of the 54 Salmonella isolates, 38.8% (21/54 were serotyped as S. typhimurium; 35.1% (19/54 as S. enterica subsp. enterica; 9.2% (5/54 as S. enterica subsp. salamae; 9.2% (5/54 as S. enterica subsp. arizona; and 7.4% (4/54 as S. enterica subsp. houtenae. Female (28/80, 35% and male (26/74, 35.1% turtles showed equal incidence of Salmonella spp. contamination. Resistance was mostly observed against ampicillin (37% followed by tetracycline (33.3%, nalidixic acid (7.4%, ciprofloxacin (5.5%, and cotrimoxazole (3.7%. The highest susceptibility was observed against gentamicin (100% and trimethoprim (98.1%. Conclusion: Our findings confirmed that people who are in close exposure to Caspian pond turtles and their feces are at the risk of Salmonella contamination. Accordingly, fundamental principles of hygiene should be applied in human contact with Caspian pond turtles. Furthermore, people should be educated about the Salmonella contamination which may occur through Caspian pond turtles.
Full Text Available Abstract Background Infections due to nontyphoidal Salmonella have increased worldwide over the last couple of decades. Salmonella enterica serotype Blockley (Salmonella Blockley infections is associated with chickens and is a rarely isolated serotype in human infections in most countries. Case presentation We report a case of human infections due to Salmonella Blockley in KwaZulu-Natal, South Africa in 2011. Three African males (aged 4, 14 and 16 presented to a clinic with diarrhoea, stomach cramps and headache. They started experiencing signs of illness a day after they consumed a common meal, consisting of meat, rice and potatoes. Stool specimens from the patients cultured Salmonella Blockley. The strains showed an indistinguishable pulsed-field gel electrophoresis pattern. Conclusion This is the first recorded case of human infections due to Salmonella Blockley in South Africa.
Introduction. The human restricted bacteria, Salmonella enterica serovar. Typhi is the major cause of typhoid fever (or enteric fever), a characteristic severe systemic illness . In 2010, typhoid fever accounted for an estimated global burden of. 27 million new cases and 200,000 deaths . For over two decades, S. enterica ...
Full Text Available Objective. The aim of this study was to establish a baseline titre for the population of Mumbai, Maharashtra, India. Method. Four hundred healthy blood donors, attending blood donation camps, were screened using a survey questionnaire. Widal tube agglutination test was performed on the diluted sera (with 0.9% normal saline of blood donors, with final dilution ranging from 1 : 40 to 1 : 320. Results. Out of 400 individuals providing samples, 78 (19.5% individuals showed antibody titres ≥ 1 : 40 for at least one antigen and 322 (80.5% showed no agglutination. The baseline antibody titres against O antigen and H antigen of Salmonella enterica serotype Typhi were found to be 1 : 40 and 1 : 80, respectively. Similarly, the baseline antibody titres for the H antigen of Salmonella enterica serotypes Paratyphi A and Paratyphi B were found to be 1 : 40 and 1 : 80, respectively. Conclusion. Thus, it was noted that the diagnostically significant cutoff of antibody titre from acute phase sample was ≥ 1 : 80 for S. Typhi O antigen and titre of ≥ 1 : 160 for both S. Typhi H antigen and S. Paratyphi BH antigen. Antibody titre of ≥ 1 : 80 can be considered significant for S. Paratyphi AH antigen.
Adriano Savoia Morales
Full Text Available Reports about acquired resistance to colistin in different bacteria species are increasing, including E. coli of animal origin, but reports of resistance in wild S. enterica of different serotypes from swine are not found in the literature. Results obtained with one hundred and twenty-six E. coli strains from diseased swine and one hundred and twenty-four S. enterica strains from diseased and carrier swine showed a frequency of 6.3% and 21% of colistin-resistant strains, respectively. When comparing the disk diffusion test with the agar dilution test to evaluate the strains, it was confirmed that the disk diffusion test is not recommended to evaluate colistin resistance as described previously. The colistin MIC 90 and MIC 50 values obtained to E. coli were 0.25 μg/mL and 0.5 μg/mL, the MIC 90 and MIC 50 to S. enterica were 1 μg/mL and 8 μg/mL. Considering the importance of colistin in control of nosocomial human infections with Gram-negative multiresistant bacteria, and the large use of this drug in animal production, the colistin resistance prevalence in enterobacteriaceae of animal origin must be monitored more closely.
Morales, Adriano Savoia; Fragoso de Araújo, Juliana; de Moura Gomes, Vasco Túlio; Reis Costa, Adrienny Trindade; dos Prazeres Rodrigues, Dália; Porfida Ferreira, Thais Sebastiana; de Lima Filsner, Pedro Henrique Nogueira; Felizardo, Maria Roberta; Micke Moreno, Andrea
Reports about acquired resistance to colistin in different bacteria species are increasing, including E. coli of animal origin, but reports of resistance in wild S. enterica of different serotypes from swine are not found in the literature. Results obtained with one hundred and twenty-six E. coli strains from diseased swine and one hundred and twenty-four S. enterica strains from diseased and carrier swine showed a frequency of 6.3% and 21% of colistin-resistant strains, respectively. When comparing the disk diffusion test with the agar dilution test to evaluate the strains, it was confirmed that the disk diffusion test is not recommended to evaluate colistin resistance as described previously. The colistin MIC 90 and MIC 50 values obtained to E. coli were 0.25 μg/mL and 0.5 μg/mL, the MIC 90 and MIC 50 to S. enterica were 1 μg/mL and 8 μg/mL. Considering the importance of colistin in control of nosocomial human infections with Gram-negative multiresistant bacteria, and the large use of this drug in animal production, the colistin resistance prevalence in enterobacteriaceae of animal origin must be monitored more closely.
Isolamento de Salmonella enterica em gambás (Didelphis aurita e Didelphis albiventris do Estado de São Paulo, Brasil Isolation of Salmonella enterica in opossum (Didelphis aurita and Didelphis albiventris of the São Paulo State, Brazil
Renata Assis Casagrande
posterior streaking in XLT4 agar. The characterization of the isolates was done through biochemical tests and serotyping. Salmonella enterica was found in 17.0% (18/106 of the D. aurita; 50% presented the bacteria in the small intestine (SI, 88.9% in the large intestine (LI and 66.7% in the cloaca. Of the S. enterica were found the subspecies: diarizonae (11.1%, enterica and houtenae (5.5% each; and the serotypes of the S. enterica enterica were Newport (83.3%, Typhimurium and Cerro (5.5% each. In the D. albiventris 17.5% (7/40 were positive; 42.8% in the SI, 85.7% in the LI and 71.4% in the cloaca. Newport (71.4% was also the most frequent serotype and the second were Typhimurium, Bareilly and Thompson (14.3% each. The presence of Salmonella enterica in the intestines of opossums in Brazil was proved.
Acar, Sinem; Bulut, Ece; Durul, Bora; Uner, Ilhan; Kur, Mehmet; Avsaroglu, M Dilek; Kirmaci, Hüseyin Avni; Tel, Yasar Osman; Zeyrek, Fadile Y; Soyer, Yesim
192 Food samples (commonly consumed 8 food types), 355 animal samples (animal feces of bovine, ovine, goat and chicken) and 50 samples from clinical human cases in Sanliurfa city, Turkey in a year were collected to determine the Salmonella enterica subsp. enterica mosaic in Turkey. 161 Salmonella isolates represented 17 serotypes, 20 sequence types (STs) and 44 PFGE patterns (PTs). 3 serotypes, S. Enteritidis, S. Typhimurium and S. Kentucky, were recovered from three different hosts. The highest discriminatory power was obtained by PFGE (SID=0.945), followed by MLST (SID=0.902) and serotyping (SID=0.885) for all isolates. The prevalence of antimicrobial resistance genes (aadA1, aadA2, strA, strB, aphA 1-Iab , bla TEM-1 , bla PSE-1 , tetA) was highly correlated with phenotypic profiles of aminoglycoside, ß-lactam and tetracycline groups (kappa >0.85). From our knowledge, this is the first study reporting spatial and temporal distribution of Salmonella species through phenotypic and genetic approaches over farm to fork chain in Turkey. Thus, our data provided further information for evolution, ecology and transmission of Salmonella in Turkey. Copyright Â© 2016. Published by Elsevier B.V.
Roer, Louise; Hendriksen, Rene S.; Leekitcharoenphon, Pimlapas
Salmonella enterica subsp. enterica bacteria are highly diverse foodborne pathogens that are subdivided into more than 1,500 serovars. The diversity is believed to result from mutational evolution, as well as intra- and interspecies recombination that potentially could be influenced by restrictio...... to the conjugational mode of horizontal gene transfer in Salmonella. Thus, we conclude that other factors must be involved in shaping the evolution of bacteria.......Salmonella enterica subsp. enterica bacteria are highly diverse foodborne pathogens that are subdivided into more than 1,500 serovars. The diversity is believed to result from mutational evolution, as well as intra- and interspecies recombination that potentially could be influenced by restriction...
Eibach, Daniel; Al-Emran, Hassan M; Dekker, Denise Myriam; Krumkamp, Ralf; Adu-Sarkodie, Yaw; Cruz Espinoza, Ligia Maria; Ehmen, Christa; Boahen, Kennedy; Heisig, Peter; Im, Justin; Jaeger, Anna; von Kalckreuth, Vera; Pak, Gi Deok; Panzner, Ursula; Park, Se Eun; Reinhardt, Alexander; Sarpong, Nimako; Schütt-Gerowitt, Heidi; Wierzba, Thomas F; Marks, Florian; May, Jürgen
Salmonella ranks among the leading causes of bloodstream infections in sub-Saharan Africa. Multidrug resistant typhoidal and nontyphoidal Salmonella (NTS) isolates have been previously identified in this region. However, resistance to ciprofloxacin has rarely been reported in West Africa. This study aims to assess susceptibility against ciprofloxacin in Salmonella causing invasive bloodstream infections among children in rural Ghana. From May 2007 until May 2012, children attending a rural district hospital in central Ghana were eligible for recruitment. Salmonella enterica isolated from blood cultures were assessed for ciprofloxacin susceptibility by Etest (susceptible minimum inhibitory concentration [MIC] ≤ 0.06 µg/mL). The gyrA, gyrB, parC, and parE genes were sequenced to identify mutations associated with changes in susceptibility to fluoroquinolones. Two hundred eighty-five Salmonella enterica isolates from 5211 blood cultures were most commonly identified as Salmonella enterica serovar Typhimurium (n = 129 [45%]), Salmonella enterica serovar Typhi (n = 89 [31%]), Salmonella enterica serovar Dublin (n = 20 [7%]), and Salmonella enterica serovar Enteritidis (n = 19 [7%]). All S. Typhi and S. Dublin were susceptible to ciprofloxacin. Reduced susceptibility (MIC >0.06 µg/mL) was found in 53% (10/19) of S. Enteritidis and in 2% (3/129) of S. Typhimurium isolates. Sequencing detected a single gyrB mutation (Glu466Asp) and a single gyrA mutation (Ser83Tyr) in all 3 S. Typhimurium isolates, while 9 of 10 S. Enteritidis harbored single gyrA mutations (Asp87Gly, Asp87Asn, or Asp87Tyr). No mutations were found in the parC and parE genes. Ciprofloxacin susceptibility in invasive NTS in rural Ghana is highly dependent on serotype. Although reduced ciprofloxacin susceptibility is low in S. Typhimurium, more than half of all S. Enteritidis isolates are affected. Healthcare practitioners in Ghana should be aware of potential treatment failure in patients with invasive
Hendriksen, Rene S.; Leekitcharoenphon, Pimlapas; Lukjancenko, Oksana; Hasman, Henrik; Aarestrup, Frank M.
ABSTRACT Salmonella enterica subsp. enterica bacteria are highly diverse foodborne pathogens that are subdivided into more than 1,500 serovars. The diversity is believed to result from mutational evolution, as well as intra- and interspecies recombination that potentially could be influenced by restriction-modification (RM) systems. The aim of this study was to investigate whether RM systems were linked to the evolution of Salmonella enterica subsp. enterica. The study included 221 Salmonella enterica genomes, of which 68 were de novo sequenced and 153 were public available genomes from ENA. The data set covered 97 different serovars of Salmonella enterica subsp. enterica and an additional five genomes from four other Salmonella subspecies as an outgroup for constructing the phylogenetic trees. The phylogenetic trees were constructed based on multiple alignment of core genes, as well as the presence or absence of pangenes. The topology of the trees was compared to the presence of RM systems, antimicrobial resistance (AMR) genes, Salmonella pathogenicity islands (SPIs), and plasmid replicons. We did not observe any correlation between evolution and the RM systems in S. enterica subsp. enterica. However, sublineage correlations and serovar-specific patterns were observed. Additionally, we conclude that plasmid replicons, SPIs, and AMR were all better correlated to serovars than to RM systems. This study suggests a limited influence of RM systems on the evolution of Salmonella enterica subsp. enterica, which could be due to the conjugational mode of horizontal gene transfer in Salmonella. Thus, we conclude that other factors must be involved in shaping the evolution of bacteria. IMPORTANCE The evolution of bacterial pathogens, their plasticity and ability to rapidly change and adapt to new surroundings are crucial for understanding the epidemiology and public health. With the application of genomics, it became clear that horizontal gene transfer played a key role in
Isolation of Salmonella enterica and serologic reactivity to Leptospira interrogans in opossums (Didelphis virginiana from Yucatán, México Aislamiento de Salmonella enterica y reactividad serológica a Leptospira interrogans en tlacuaches (Didelphis virginiana de Yucatán, México
Hugo Antonio RUIZ-PIÑA
Full Text Available 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.La presencia de Salmonella enterica y evidencia serológica de infección por Leptospira interrogans fueron detectadas en tlacuaches de la especie Didelphis virginiana capturados en una localidad semi-urbana del estado de Yucatán, México. Se capturaron 91 marsupiales durante el período de abril de 1996 a mayo de 1998. De un total de 17 muestras de heces, se aislaron cuatro serotipos de Salmonella enterica subsp. enterica (Sandiego, Newport, Anatum y Minnesota y una Salmonella enterica subsp. arizonae serovar O44:Z4,Z23:-. En algunos tlacuaches se registraron infecciones mixtas. De 81 muestras de suero, cuatro (4,9% presentaron reacciones positivas con los serovares pomona y wolffi, ambos pertenecientes al género Leptospira. Los tlacuaches con serología positiva fueron capturados en el hábitat peridomiciliar. Los animales infectados con Salmonella enterica fueron capturados en los períodos de seca y
A ciprofloxacin resistant (CipR) Salmonella enterica subsp. enterica serovar Kentucky ST198 has rapidly and extensively disseminated globally to become a major food-safety and public health concern. Here, we report a complete genome sequence of a CipR S. Kentucky ST198 strain PU131 isolated from a ...
Batista, Diego Felipe Alves; de Freitas Neto, Oliveiro Caetano; Lopes, Priscila Diniz; de Almeida, Adriana Maria; Barrow, Paul Andrew; Berchieri, Angelo
Salmonella Pullorum and Salmonella Gallinarum are classified as biovars of Salmonella enterica subsp. enterica serovar Gallinarum. These salmonellae are the causative agents of Pullorum disease and fowl typhoid, respectively, and are widely distributed throughout the world. Although many developed countries have eradicated these diseases from commercial poultry, they are still the cause of significant economic loss in developing countries. When serovar Gallinarum is isolated, it is difficult to immediately differentiate between biovars because they are antigenically identical by serotyping. However, they cause distinct diseases with different epidemiology, and therefore it is important to differentiate them. This may be done biochemically but takes 2 to 3 days. In the present study, S. Pullorum and S. Gallinarum whole genomes were compared, and 1 genomic region of difference, which is part of the ratA gene, was chosen as a molecular marker for a polymerase chain reaction assay to differentiate rapidly between these organisms. In all, 26 strains of S. Gallinarum and 17 S. Pullorum strains were tested and successfully differentiated by the assay.
Full Text Available ABSTRACT Salmonella enterica is a large group of Gram-negative bacteria responsible for a number of foodborne infections associated with the consumption of contaminated poultry products. The hygienic status of raw chicken meat marketed at Ibague, Tolima, Colombia, is currently unknown. To address this issue, a cross-sectional study was conducted to estimate the prevalence of Salmonella spp., in raw chicken marketed at different outlets in this city. Salmonella spp. was isolated by standard microbiological methods, followed by biochemical, serological, and molecular confirmation. Additionally, risk factors associated with the presence of the bacteria were identified. The prevalence of Salmonella in raw chicken was 17.41% (47/270, and 14 different serotypes were found, out of which S. Paratyphi B (36.17%, S. Hvittingfoss (19.15% and S. Muenster (10.64% were the most prevalent and represented 65.95% of all serotypes. Amplification of 284 bp of the invA gene was achieved by PCR in a number of randomly selected isolates. Raw chicken as the only type of meat sold at stores (odds ratio: 2,157, p<0.05, and stainless steel as a contact surface of chicken meat (odds ratio: 13,29, p<0.05, were found to be potential risk factors for the presence of Salmonella in chicken meat. This work serves as a reference about the current status of Salmonella in chicken meat marketed in Ibague, Tolima, Colombia, and indicates the need to establish appropriate control and contingency measures to minimize the presence of the bacteria in raw chicken to prevent its transmission to humans.
Hokunan, Hidekazu; Koyama, Kento; Hasegawa, Mayumi; Kawamura, Shuso; Koseki, Shigenobu
We investigated the survival kinetics of Salmonella enterica and enterohemorrhagic Escherichia coli under various water activity (a w ) conditions to elucidate the net effect of a w on pathogen survival kinetics and to pursue the development of a predictive model of pathogen survival as a function of a w . Four serotypes of S. enterica (Stanley, Typhimurium, Chester, and Oranienburg) and three serotypes of enterohemorrhagic E. coli ( E. coli O26, E. coli O111, and E. coli O157:H7) were examined. These bacterial strains were inoculated on a plastic plate surface at a constant relative humidity (RH) (22, 43, 58, 68, or 93% RH, corresponding to the a w ) or on a surface of almond kernels (a w 0.58), chocolate (a w 0.43), radish sprout seeds (a w 0.58), or Cheddar cheese (a w 0.93) at 5, 15, or 25°C for up to 11 months. Under most conditions, the survival kinetics were nonlinear with tailing regardless of the storage a w , temperature, and bacterial strain. For all bacterial serotypes, there were no apparent differences in pathogen survival kinetics on the plastic surface at a given storage temperature among the tested RH conditions, except for the 93% RH condition. Most bacterial serotypes were rapidly inactivated on Cheddar cheese when stored at 5°C compared with their inactivation on chocolate, almonds, and radish sprout seeds. Distinct trends in bacterial survival kinetics were also observed between almond kernels and radish sprout seeds, even though the a w s of these two foods were not significantly different. The survival kinetics of bacteria inoculated on the plastic plate surface showed little correspondence to those of bacteria inoculated on food matrices at an identical a w . Thus, these results demonstrated that, for low-a w foods and/or environments, a w alone is insufficient to account for the survival kinetics of S. enterica and enterohemorrhagic E. coli .
Tajbakhsh, Mercedeh; Avini, Mohammad Yaghoobi; Alikhajeh, Jahan; Tajeddin, Elahe; Rahbar, Mohammad; Eslami, Parisa; Alebouyeh, Masoud; Zali, Mohammad Reza
Broad-spectrum β-lactams are used for empirical therapy of severe infections with non-typhoid Salmonella serotypes; however, activities of these drugs against the strains producing different β-lactamase is not so clear. This study investigated the prevalence of β-lactamase genes among isolates of S. enterica serovars from human faecal samples and determined their diversity in activity against different β-lactams. Antimicrobial resistance of faecal isolates of S. enterica to extended-spectrum cephalosporins was analysed and MIC values were determined for the strains presenting extended-spectrum β-lactamases (ESBLs) phenotypes. The β-lactamase genes were identified by PCR and sequencing. β-lactamase activity of the Salmonella strains exhibiting ESBL phenotype was detected by biological, iodometric, spectrophotometry and nitrocefin assays. Out of 202 S. enterica isolates, ESBLs phenotype was detected among 3.4% (7/202) of the strains. blaTEM-1 and blaCTX-M-15 were among the frequent β-lactamase genes. Detection of blaTEM-169 in S. enterica serovar Typhimurium and S. enterica serovar Bredeney and blaPER-1 in S. enterica serovar Infantis was a new finding in this experiment. Location of blaCTX-M-15/blaTEM-169/blaPER-1 genes on plasmid was confirmed in a transformation experiment. While crude extracts of the enzymes from each strain showed higher activity against cephalothin and cefotaxime, the lowest activity was detected against ceftazidime. The greatest synergistic activity was seen in a strain of S. enterica that carried blaCTX-M-15 and blaPER-1 genes compared with those presenting blaCTX-M-15/blaTEM-169 or blaCTX-M-15/blaTEM-1 genotypes. The results show dissemination of ESBLs encoding genes and their combined activity among different serovars of S. enterica that are a threat for future treatment options.
Potnis, Neha; Soto-Arias, José Pablo; Cowles, Kimberly N.; van Bruggen, Ariena H. C.; Jones, Jeffrey B.
Salmonella enterica rarely grows on healthy, undamaged plants, but its persistence is influenced by bacterial plant pathogens. The interactions between S. enterica, Xanthomonas perforans (a tomato bacterial spot pathogen), and tomato were characterized. We observed that virulent X. perforans, which establishes disease by suppressing pathogen-associated molecular pattern (PAMP)-triggered immunity that leads to effector-triggered susceptibility, created a conducive environment for persistence of S. enterica in the tomato phyllosphere, while activation of effector-triggered immunity by avirulent X. perforans resulted in a dramatic reduction in S. enterica populations. S. enterica populations persisted at ∼10 times higher levels in leaves coinoculated with virulent X. perforans than in those where S. enterica was applied alone. In contrast, S. enterica populations were ∼5 times smaller in leaves coinoculated with avirulent X. perforans than in leaves inoculated with S. enterica alone. Coinoculation with virulent X. perforans increased S. enterica aggregate formation; however, S. enterica was not found in mixed aggregates with X. perforans. Increased aggregate formation by S. enterica may serve as the mechanism of persistence on leaves cocolonized by virulent X. perforans. S. enterica association with stomata was altered by X. perforans; however, it did not result in appreciable populations of S. enterica in the apoplast even in the presence of large virulent X. perforans populations. Gene-for-gene resistance against X. perforans successively restricted S. enterica populations. Given the effect of this interaction, breeding for disease-resistant cultivars may be an effective strategy to limit both plant disease and S. enterica populations and, consequently, human illness. PMID:24632252
Traoré, Oumar; Nyholm, Outi; Siitonen, Anja; Bonkoungou, Isidore Juste O; Traoré, Alfred S; Barro, Nicolas; Haukka, Kaisa
This study investigated the prevalence, serotypes and antimicrobial sensitivity patterns of Salmonella enterica in environment in Ouagadougou, Burkina Faso. A total of 476 samples, consisting of 36 samples of tap water, 51 samples of well water, 87 samples of channel water, 44 samples of reservoir water, 238 samples of fish, and 20 samples of lettuce were examined using standard bacteriological procedures for Salmonella. Salmonella were isolated from 98 samples. Salmonella were rare in drinking water, since they were not found at all from the tap water, and only in 2 % of well water. Salmonella were more common in the water of reservoir of Tanghin (15 %), reservoir of Yamtenga (20 %), and in the water channels in the city (from 20 to 31 %). Salmonella were commonly isolated from the fish (24 %) caught from the reservoir of Tanghin and from the lettuce (50 %) irrigated with water from Tanghin. The Salmonella isolates were found to represent 50 different serotypes. The 11 most common serotypes were Salmonella Bredeney and S. Colindale (both 8.2 %), S. Muenster (6.1 %), S. Korlebu (5.1 %), S. Eastbourne and S. Poona (both 4.1 %), and S. Agona, S. Derby, S. Drac, S. Senftenberg, S. Waycross (each 3.1 %), accounting for 51.3 % of all the isolates. In general, the Salmonella strains were sensitive to the antimicrobials tested, but two strains were resistant to streptomycin and many more intermediate to streptomycin or sulphonamide. This study highlights the common prevalence of Salmonella and the high diversity of Salmonella serotypes in aquatic environment in Ouagadougou, Burkina Faso. Therefore, various human activities linked to water and consumption of water-related products, such as fish and lettuce, can lead to human Salmonella infections.
Ktsoyan, Zhanna A.; Mkrtchyan, Mkhitar S.; Zakharyan, Magdalina K.
with the comparative counts of cultivated bacteria showed an inverse relationship with the IgE concentration. Earlier we reported the elevated level of IL-17 in patients infected by S. Enteritidis. In the current study a significant correlation was found between the concentrations of IL-17 and IgE suggesting...
Šplíchal, Igor; Trebichavský, Ilja; Muneta, Y.; Mori, Y.
Roč. 33, - (2002), s. 291-297 ISSN 0928-4249 R&D Projects: GA ČR GA524/01/0917 Grant - others:Ministry of Agriculture Forestry and Fisheries(JP) RCP3260 Keywords : typhimurium * gnotobiotic piglet Subject RIV: EE - Microbiology, Virology Impact factor: 2.031, year: 2002
Full Text Available Background: The emergence of antimicrobial-resistant bacteria with biofilm formation ability may be a major threat to public health and food safety and sanitation. Objectives: The aim of this study was to determine antibiotic resistance patterns and biofilm production characteristics of Salmonella typhimurium isolated from different species of birds. Materials and Methods: The antibiotic resistance patterns of 38 pre-identified isolates were screened by standard Kirby-Bauer disc-diffusion method performed on Mueller–Hinton agar to a panel of 17 antibiotics. The extent of biofilm formation was measured by Microtiter plate (MTP-based systems. Results: The highest antimicrobial resistance was detected against nalidixic acid (97%, followed by doxycycline (86%, colistin (84%, streptomycin (84% and tetracycline (84%. All isolates were sensitive to amikacin (100% and 97% and 95% of the isolates were sensitive to ceftazidime and ceftriaxone, respectively. Twenty one different antibiotic resistance patterns were observed among S. typhimurium isolates. According to the results of the microtitre plate biofilm assay, there was a wide variation in biofilm forming ability among S. typhimurium isolates. Most of the isolates (60.52% were not capable of producing biofilm, while 26.31%, 7.89%, and 5.26% isolates were weak, strong and moderate biofilm producers, respectively. Conclusions: It was concluded that nearly all S. typhimurium isolates revealed a high multiple antibiotic resistant with low biofilm forming capabilities which proposed low association between biofilm formation and antibiotic resistance of a major food important pathogen.
Pantip, Sirichote; Aroon, B.; Kanokwan, Tienmanee
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...
Le Hello, Simon; Hendriksen, Rene S.; Doublet, Benoit
selected in 3 steps in Egypt during the 1990s and the early 2000s and has now spread to several countries in Africa and, more recently, in the Middle East. Poultry has been identified as a potential major vehicle for infection by this clone. Continued surveillance and appropriate control measures should...
Molbak, K.; Baggesen, Dorte Lau; Aarestrup, Frank Møller
, and tetracycline. An increasing proportion of DT104 isolates also have reduced susceptibility to fluoroquinolones. Methods The Danish salmonella surveillance program determines the phage types of all typhimurium strains from the food chain, and in the case of suspected outbreaks, five-drug-resistant strains...... findings here. Results Until 1997, DT104 infections made up less than 1 percent of all human salmonella infections. The strain isolated from patients in the first community outbreak of DT104 in Denmark, in 1998, was resistant to nalidixic acid and had reduced susceptibility to fluoroquinolones...... with fluoroquinolones. Conclusions Our investigation of an outbreak of DT104 documented the spread of quinolone-resistant bacteria from food animals to humans; this spread was associated with infections that were difficult to treat. Because of the increase in quinolone resistance in salmonella, the use...
Full Text Available In tandem to the Salmonellosis infection worldwide, a study was conducted to determine the conserved and nonconserved region in Salmonella enterica sub-species so that it can be used to design probes in biosensors for the detection of Salmonella enterica as a species or sub-species distinctively. The region was selected for this study through fliC gene that is present in all Salmonella sub-species, encodes the Salmonella flagella or flagellin determines the serotype due to its H antigen and provides virulence to the bacteria. All sub-species were analyzed in a group of six and then analyzed individually in groups of two with all possible combinations to determine the overlapping regions. Based on the study, the predominant conserved region for S. enterica sub-species is between 103 and 158, whereas the non-conserved region is from 1245 to 1285. A phylogenetic tree was constructed for the S. enterica sub-species to determine the evolution of the Salmonella sub-species.
Full Text Available Abstract Salmonella enterica serovar Typhimurium and S.enterica serovar Enteritidis are the most frequently isolated serovars from food-borne diseases throughout the world. According to their antigenic profiles, salmonella shows different disease syndromes and host specificities. It is necessary and important to discriminate salmonella serovars from each other in order to ensure that each pathogen and its epidemiology are correctly recognized. Many PCR-based methods have been developed to identify salmonella serovars. The objective of present study was to identify S. Typhimurium in avians from different regions including: North, Northwest and capital city (Tehran of Iran. Also in this research, the quality of CHROMagar™ Salmonella medium (CAS medium in veterinary medicine was evaluated. The results of present study showed that out of 1870 intestine samples, fifty two S. Typhimurium including broiler (n=13, layer (n=12, duck (n=5, goose (n=5, sparrow (n=8, canary (n=3, pigeon (n=5 and African grey parrot (n=1 were identified using serotyping as well as multiplex-PCR. In conclusion, important measures must be taken on prevention and propagation of S. Typhimurium among avians. CHROMagar™ Salmonella medium has high levels of sensitivity and specificity and reduced the time to final identification of salmonella spp. in comparison with biochemical tests.
Gebreyes, Wondwossen Abebe; Altier, Craig
As part of a longitudinal study of antimicrobial resistance among salmonellae isolated from swine, we studied 484 Salmonella enterica subsp. enterica serovar Typhimurium (including serovar Typhimurium var. Copenhagen) isolates. We found two common pentaresistant phenotypes. The first was resistance to ampicillin, chloramphenicol, streptomycin, sulfamethoxazole, and tetracycline (the AmCmStSuTe phenotype; 36.2% of all isolates), mainly of the definitive type 104 (DT104) phage type (180 of 187 ...
Rezende, Ana Carolina B; Crucello, Juliana; Moreira, Rafael C; Silva, Beatriz S; Sant'Ana, Anderson S
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
Zhou, L.; Jones, S.C.P.; Angen, Øystein
, but the method has liminations, for example, cross-reactions between serotypes 3, 6, and 8. This study describes the development of a serotype 3-specific PCR, based on the capsule locus, which can be used in a multiplex format with the organism's specific gene apxIV. The PCR test was evaluated on 266 strains...
Dubreuil, J.D.; Letellier, A.; Stenbæk, Eva
A polystyrene agglutination test has been developed for serotyping Actinobacillus pleuropneumoniae serotype 5a and 5b strains. Protein A-coated polystyrene microparticles were sensitized with a murine monoclonal antibody recognizing an epitope on serotype 5 LPS-O chain as shown by SDS-PAGE and We......A polystyrene agglutination test has been developed for serotyping Actinobacillus pleuropneumoniae serotype 5a and 5b strains. Protein A-coated polystyrene microparticles were sensitized with a murine monoclonal antibody recognizing an epitope on serotype 5 LPS-O chain as shown by SDS...... suspension of bacterial cells grown for 18 h. All A, pleuropneumoniae strains had been previously serotyped using standard procedures, The polystyrene agglutination test was rapid (less than 3 min) and easy to perform. Overall a very good correlation (97.3%) with the standard techniques was found...
Multiplex polymerase chain reaction (PCR) was used for molecular typing of Salmonella enterica serovars in Egypt. During the summer of 2010, a total of 1075 samples were collected from cattle, sheep and poultry farms to be subjected for isolation of Salmonella (290 rectal swabs from cattle, 335 rectal swabs from sheep ...
Bonardi, Silvia; Alpigiani, Irene; Bruini, Ilaria; Barilli, Elena; Brindani, Franco; Morganti, Marina; Cavallini, Pierugo; Bolzoni, Luca; Pongolini, Stefano
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,,12:i:- (17.5%) and Salmonella Rissen (10.0%). In faecal samples S. Derby was prevalent (51.4%), followed by S. enterica 4,, 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,,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,,12:i:-, S. Rissen, Salmonella
Rocke, Tonie E.; Smith, Susan R.; Miyamoto, Amy; Shadduck, Daniel J.
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.
João B.P. Guerra Filho
Full Text Available ABSTRACT: Salmonellosis is a foodborne disease caused by bacteria of the genus Salmonella, being pigs and pork-products potentially important for its occurrence. In recent decades, some serovars of Salmonella have shown increase of resistance to conventional antimicrobials used in human and animal therapy, with serious risks for public health. The aim of this study was to evaluate feces (n=50, mediastinal (n=50, mesenteric (n=50 and mandibular (n=50 lymph nodes obtained from slaughter houses for Salmonella spp. Positive samples were serotyped and subjected to an in vitro antimicrobial susceptibility test, including the extended-spectrum beta-lactamase (ESBL production. Salmonella species were identified in 10% (20/200 of total samples. From these, 20% (10/50 were identified in the submandibular lymph nodes, 18% (9/50 in the mesenteric lymph nodes, 2% (1/50 in feces and 0% (0/50 in the mediastinal lymph nodes. The serotypes found were Salonella Typhimurium (55%, S. enterica subsp. enterica 4,5,12: i: - (35%, S. Brandenburg and S. Derby with 5% (5% each. All strains showed resistance to at least one antimicrobial; 90% were resistant to four or more antimicrobials, and 15% were multidrug-resistant. Resistance to ciprofloxacin, tetracycline and nalidixic acid was particularly prevalent amongst the tested serovars. Here, we highlighted the impact of pigs in the epidemiological chain of salmonellosis in domestic animals and humans, as well as the high antimicrobial resistance rates of Salmonella strains, reinforcing the necessity for responsible use of antimicrobials for animals as an emergent One Health issue, and to keep these drugs for human therapy approaches.
Jacqueline Boldrin de Paiva
Full Text Available 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 B12 in 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.
Full Text Available The present study reports the isolation of Salmonella enterica in organs of free-living domestic pigeons. In the clinic examination, the presence of feces in the peri-cloacal and abdominal regions were observed, as well as symptoms such as cachexy, incoordination and opisthotonos. Before any therapeutic protocol was applied the bird died and a necropsy was then performed for the removal of spleen, liver, kidney and intestine for bacteriological examination and antibiotic sensitivity test. Salmonella enterica subsp.enterica (O:4,5:i- and Salmonella enterica subsp. enterica serovar Typhimurium were isolated from the liver and intestine and the sensitivity test demonstrated that these strains are sensitive to several antibiotics.
Lu, Po-Liang; Hwang, In-Jane; Tung, Ya-Lina; Hwang, Shang-Jyh; Lin, Chun-Lu; Siu, LK
Background An increase in the number of attendees due to acute gastroenteritis and fever was noted at one hospital emergency room in Taiwan over a seven-day period from July to August, 2001. Molecular and epidemiological surveys were performed to trace the possible source of infection. Methods An epidemiological investigation was undertaken to determine the cause of the outbreak. Stool and blood samples were collected according to standard protocols per Center for Disease Control, Taiwan. Typing of the Salmonella isolates from stool, blood, and food samples was performed with serotyping, antibiotypes, and pulsed field gel electrophoresis (PFGE) following XbaI restriction enzyme digestion. Results Comparison of the number of patients with and without acute gastroenteritis (506 and 4467, respectively) during the six weeks before the outbreak week revealed a significant increase in the number of patients during the outbreak week (162 and 942, respectively) (relative risk (RR): 1.44, 95% confidence interval (CI): 1.22–1.70, P value bakery and were distributed to six different traditional Chinese markets., Salmonella enterica subsp. enterica serovar Enteritidis (S. Enteritidis) was isolated from the stool samples of 28 of 32 individuals and from a recalled bread sample. All S. Enteritidis isolates were of the same antibiogram. PFGE typing revealed that all except two of the clinical isolates and the bread isolates were of the same DNA macrorestriction pattern. Conclusions The egg-covered bread contaminated with S. Enteritidis was confirmed as the vehicle of infection. Alertness in the emergency room, surveillance by the microbiology laboratory, prompt and thorough investigation to trace the source of outbreaks, and institution of appropriate control measures provide effective control of community outbreaks. PMID:15541186
Hernández, Jorge; Lindberg, Peter; Waldenström, Jonas; Drobni, Mirva; Olsen, Björn
A novel Salmonella serovar was isolated from Peregrine falcon (Falco peregrinus) nestlings in northern Sweden in 2006. Three isolates of the same clone was retrieved from three falcon siblings and characterized as Salmonella enterica sub-species enterica: O-phase 13, 23:-: e, n, z 15 and the H-phase was not present. We propose the geographical name Salmonella enterica, sub-species entericaserovar Pajala to this novel Salmonella.Keywords: Salmonella; epidemiology; ecology; peregrine falcon; no...
Leonard, Susan R; Lacher, David W; Lampel, Keith A
Classical bacteriological characteristics of Salmonella enterica indicate that the members of this species are unable to utilize lactose as a carbon source. However, lactose-fermenting (Lac+) strains of several Salmonella serovars have been isolated from different foodborne outbreaks as well as different geographical regions worldwide. In the present study, we sequenced the genomes of 13 Lac + S. enterica isolates and characterized the lac region, comparing it to the lac region in other enteric bacterial species. Genetic analysis of the lac operons in the S. enterica genomes revealed that they all contain intact lacI, lacZ, and lacY genes. However, lacA was truncated in all of the S. enterica subsp. enterica isolates, encoding a 56 amino acid peptide rather than the full length 220 amino acid LacA protein. Molecular analyses of the 13 isolates revealed that the lac operon resided on a plasmid in some strains and in others was integrated into the bacterial chromosome. In most cases, an insertion sequence flanked at least one end of the operon. Interestingly, the S. enterica Montevideo and S. enterica Senftenberg isolates were found to harbor a plasmid with a high degree of sequence similarity to a plasmid from Klebsiella pneumoniae strain NK29 that also harbors the lac operon. In addition, two S. enterica Tennessee isolates carried two copies of the lac operon. Phylogenetic analysis based on lacIZY gene sequences determines distinct clusters, and reveals a greater correlation between lacIZY sequence and flanking organization than with either bacterial species or genomic location. Our results indicate that the lac region is highly mobile among Enterobacteriaceae and demonstrate that the Lac + S. enterica subsp. enterica serovars acquired the lac region through parallel events. The acquisition of the lac operon by several S. enterica serovars may be indicative of environmental adaptation by these bacteria.
Kieboom, J.; Abee, T.
Salmonella enterica serovar Typhimurium does not survive a pH 2.5 acid challenge under conditions similar to those used for Escherichia coli (J. W. Foster, Nat. Rev. Microbiol. 2:898-907, 2004). Here, we provide evidence that S. enterica serovar Typhimurium can display arginine-dependent acid
Soto-Arias, José Pablo; Groves, Russell L.
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
Nov 5, 2008 ... Typhoid fever continues to remain a health problem as the causative organism, Salmonella enterica serovar typhi, has developed resistance to many antibiotics used. This study was undertaken to determine the current pattern of resistance to antimicrobial agents by S. enterica serovar typhi isolates.
Salmonella enterica subsp. enterica serovar Enteritidis is a wide-host-range pathogen. Occasionally, it is involved in invasive infections, leading to a high mortality rate. Here, we present the draft genome sequences of four S Enteritidis strains obtained from human and avian hosts that had been involved in bacteremia, gastroenteritis, and primary infections.
Di Giannatale, E; Sacchini, L; Persiani, T; Alessiani, A; Marotta, F; Zilli, K
To provide an epidemiologic interpretation of a suspected outbreak of food poisoning caused by Salmonella enterica subspecies enterica serovar Berta strains isolated from humans and from the leftovers of the implicated foods (cream, dairy-based desserts and eggs). We have correlated the similarity between the strains through genotyping with Pulsed Field Gel Electrophoresis (PFGE), studying antimicrobial sensitivity patterns and epidemiological investigation. The clonal origin of the outbreak was confirmed by all laboratory tests. PFGE analysis of the restriction profiles obtained with XbaI and SpeI revealed a certainly correlation from the strains isolated from the various sources, while the antimicrobial sensitivity pattern was the same in all cases, with all strains sensitive to all antibiotics tested. Poor hygiene conditions in the facility concerned, lack of hygiene in food handling, high summer temperatures and positive cultures from asymptomatic staff could all be implicated in the infection, with food being the means through which it spread. This study describes the first outbreak of food poisoning caused by Salmonella enterica subspecies enterica serovar Berta (Salmonella Berta) reported in Italy. It confirms the importance of correlating epidemiological investigations with genotyping and phenotyping to understand the dynamics of infection. © 2012 The Authors. Letters in Applied Microbiology © 2012 The Society for Applied Microbiology.
Full Text Available Flavobacterium psychrophilum is a devastating bacterial pathogen of salmonids reared in freshwater worldwide. So far, serological diversity between isolates has been described but the underlying molecular factors remain unknown. By combining complete genome sequence analysis and the serotyping method proposed by Lorenzen and Olesen (1997 for a set of 34 strains, we identified key molecular determinants of the serotypes. This knowledge allowed us to develop a robust multiplex PCR-based serotyping scheme, which was applied to 244 bacterial isolates. The results revealed a striking association between PCR-serotype and fish host species and illustrate the use of this approach as a simple and cost-effective method for the determination of F. psychrophilum serogroups. PCR-based serotyping could be a useful tool in a range of applications such as disease surveillance, selection of salmonids for bacterial coldwater disease resistance and future vaccine formulation.
Dubreuil, J.D.; Letellier, A.; Stenbæk, Eva
A polystyrene agglutination test has been developed for serotyping Actinobacillus pleuropneumoniae serotype 5a and 5b strains. Protein A-coated polystyrene microparticles were sensitized with a murine monoclonal antibody recognizing an epitope on serotype 5 LPS-O chain as shown by SDS......-PAGE and Western blotting, A total of 205 A. pleuropneumoniae, strains including all 12 serotype reference strains and 13 strains representing 8 common bacterial species associated with swine or related to A, pleuropneumoniae, were tested by mixing 25 mu L of polystyrene reagent with the same volume of a dense...... suspension of bacterial cells grown for 18 h. All A, pleuropneumoniae strains had been previously serotyped using standard procedures, The polystyrene agglutination test was rapid (less than 3 min) and easy to perform. Overall a very good correlation (97.3%) with the standard techniques was found...
Stenbaek, E.I.; HovindHaugen, K.
Until now 12 serotypes of Actinobacillus pleuropneumoniae have been recognized. The specificity of the serotypes reside in the carbohydrate composition of the capsular polysaccharides and lipopolysaccharides (LPS). The LPS of A. pleuropneumoniae serotype 2 is a smooth type LPS with O......-PAGE). The MAI, 102-G02 was directed against an epitope on the O-chain of the LPS and was used to define a new LPS variant of A. pleuropneumoniae serotype 2 (referred to as A. pleuropneumoniae serotype 2X). Investigation of the reactivity of the MAb 102-G02 against an A. pleuropneumoniae serotype 2X field...
Ho, Ying-Ning; Tsai, Hsin-Chi; Hsu, Bing-Mu; Chiou, Chien-Shun
Salmonella is one of the most common pathogens of waterborne and foodborne disease-causing pathogens. In this study, we collected 172 surface water samples from Puzih River and Kaoping River between the years 2010 and 2011. Salmonella was detected in 31.7% (32/101) and 42.2% (30/71) of the samples from the two rivers, respectively. From these positive samples, 44 Salmonella isolates were obtained from these positive samples and were characterized using serotyping and pulsed-field gel electrophoresis (PFGE) genotyping. The isolates were found with 17 serovars and 32 PFGE patterns. Salmonella enterica Newport, Bareilly, Kedougou, Albany and subspecies IIIb 50:k:z were the five most common serovars in aquatic environmental Salmonella isolates. In addition, of the total clinical samples from Chiayi and Kaohsiung, 33.7% (60/178) Newport serovars were isolated. After conducting categorical analysis, we found that the serovar Newport was not uniformly distributed cross the cities. The serovar Newport was over-represented (p<0.001) among human isolates in Chiayi and Kaohsiung. To investigate the association between Salmonella isolates from aquatic environment and human samples, we compared the environmental PFGE patterns of the test samples with those of 2438 clinical isolates, obtained from 51 hospitals across the country between 2010 and 2011. Of the 32 PFGE genotypes of environmental isolates, 8 genotypes were identical to those of clinical samples. Serovar Newport isolates with PFGE patterns SNX.119 and SNX.183 obtained from Puzih River samples were also identified in human samples at a local hospital. These suggest that there is a link between environmental and human clinical Salmonella. Identification of Salmonella serovars and genotypes present in surface water provides an indication of the specific S. enterica serovars and genotypes present in humans. This is the first study to investigate the Salmonella serovars and genotypes present in aquatic environment and
Farias, L F P; Oliveira, C J B; Medardus, J J; Molla, B Z; Wolfe, B A; Gebreyes, W A
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. © 2014 Blackwell Verlag GmbH.
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
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. Copyright © 2016, American Society for Microbiology. All Rights Reserved.
Laorden, Lorena; Herrera-León, Silvia; Martínez, Ilargi; Sanchez, Ainhoa; Kromidas, Lefteris; Bikandi, Joseba; Rementeria, Aitor; Echeita, Aurora; Garaizar, Javier
We analyzed a collection of 60 Salmonella enterica 4,5,12:i:- phage type U302 multidrug-resistant monophasic variant strains, isolated in Spain between 2000 and 2007. Most strains showed resistance to ampicillin (A), chloramphenicol (C), sulfamethoxazole (Su), gentamicin (G), streptomycin (S), tetracycline (T), and co-trimoxazole (SxT) (an ACSuGSTSxT resistance pattern). Only one pulsed-field gel electrophoresis (PFGE) type was detected, with 19 subtypes (Simpson's index of diversity [SID] = 0.89). Multiple-locus variable-number tandem-repeat analysis (MLVA) showed more variability, with 32 profiles (SID = 0.97), but only showed diversity at the STTR5 and STTR6 loci. PCR and sequencing demonstrated all strains contained the same allantoin-glyoxylate pathway deletion. Four types of deletions were detected in the fljAB operon, all starting at the same position, at the STM2758 gene, and followed by an IS26 insertion. Furthermore, a representative set of strains of the four deletion types harbored plasmids with IS26. We propose that a Salmonella enterica serotype Typhimurium U302 multidrug-resistant (ACSuGSTSxT) strain, defective for the allantoin-glyoxylate pathway and containing IS26 at plasmid pU302L, could be the ancestor of the variant in Spain. PMID:20943866
Abd El Ghany, Moataz
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.
Peng Chen, Zhongxing; Morris, J Glenn; Rodriguez, Ramon L; Shukla, Aparna Wagle; Tapia-Núñez, John; Okun, Michael S
Two decades ago, botulinum neurotoxin (BoNT) type A was introduced to the commercial market. Subsequently, the toxin was approved by the FDA to address several neurological syndromes, involving muscle, nerve, and gland hyperactivity. These syndromes have typically been associated with abnormalities in cholinergic transmission. Despite the multiplicity of botulinal serotypes (designated as types A through G), therapeutic preparations are currently only available for BoNT types A and B. However, other BoNT serotypes are under study for possible clinical use and new clinical indications; To review the current research on botulinum neurotoxin serotypes A-G, and to analyze potential applications within basic science and clinical settings; The increasing understanding of botulinal neurotoxin pathophysiology, including the neurotoxin's effects on specific neuronal populations, will help us in tailoring treatments for specific diagnoses, symptoms and patients. Scientists and clinicians should be aware of the full range of available data involving neurotoxin subtypes A-G.
Benfield, Thomas; Skovgaard, Marlene; Schønheyder, Henrik Carl
There is limited knowledge of serotypes that cause non-bacteremic pneumococcal pneumonia (NBP). Here we report serotypes, their associated disease potential and coverage of pneumococcal conjugate vaccines (PCV) in adults with NBP and compare these to bacteremic pneumonia (BP).......There is limited knowledge of serotypes that cause non-bacteremic pneumococcal pneumonia (NBP). Here we report serotypes, their associated disease potential and coverage of pneumococcal conjugate vaccines (PCV) in adults with NBP and compare these to bacteremic pneumonia (BP)....
Seyfarth, Anne Mette; Wegener, Henrik Caspar; FrimodtMoller, N.
.4% of strains from cattle, 11.1% of strains from pigs and 9.2% of strains from poultry. Multiple resistance, i.e. resistance against at least four antimicrobial agents, was found in 9.2% of the human strains, but in only two of the cattle isolates, The majority of the multi-resistant strains in humans were from......: Poultry strains were usually resistant only to ampicillin, white pig and cattle isolates were most often resistant to sulphonamide, tetracycline and streptomycin. Typing of the strains showed that some animal strains and human strains were indistinguishable. In conclusion, while antimicrobial resistance......We have studied the frequency of antimicrobial resistance and epidemiological relatedness among 473 isolates of Salmonella enterica subsp, enterica serovar typhimurium (S. typhimurium) from human and veterinary sources. The human strains were clinical isolates from patients with diarrhoea sent...
Seyfarth, Anne Mette; Wegener, Henrik Caspar; FrimodtMoller, N.
infections contracted outside Denmark, most often in southern Europe or south-east Asia. Resistance in human strains was most common against tetracycline (13%), ampicillin (12%), sulphonamide (12%), streptomycin (10%) and chloramphenicol (8%). The resistance pattern differed somewhat in animal isolates......: Poultry strains were usually resistant only to ampicillin, white pig and cattle isolates were most often resistant to sulphonamide, tetracycline and streptomycin. Typing of the strains showed that some animal strains and human strains were indistinguishable. In conclusion, while antimicrobial resistance......We have studied the frequency of antimicrobial resistance and epidemiological relatedness among 473 isolates of Salmonella enterica subsp, enterica serovar typhimurium (S. typhimurium) from human and veterinary sources. The human strains were clinical isolates from patients with diarrhoea sent...
Santiviago Carlos A
Full Text Available Abstract Background The recently described Type VI Secretion System (T6SS represents a new paradigm of protein secretion in bacteria. A number of bioinformatic studies have been conducted to identify T6SS gene clusters in the available bacterial genome sequences. According to these studies, Salmonella harbors a unique T6SS encoded in the Salmonella Pathogenicity Island 6 (SPI-6. Since these studies only considered few Salmonella genomes, the present work aimed to identify novel T6SS loci by in silico analysis of every genome sequence of Salmonella available. Results The analysis of sequencing data from 44 completed or in progress Salmonella genome projects allowed the identification of 3 novel T6SS loci. These clusters are located in differentially-distributed genomic islands we designated SPI-19, SPI-20 and SPI-21, respectively. SPI-19 was identified in a subset of S. enterica serotypes including Dublin, Weltevreden, Agona, Gallinarum and Enteritidis. In the later, an internal deletion eliminated most of the island. On the other hand, SPI-20 and SPI-21 were restricted to S. enterica subspecies arizonae (IIIa serotype 62:z4,z23:-. Remarkably, SPI-21 encodes a VgrG protein containing a C-terminal extension similar to S-type pyocins of Pseudomonas aeruginosa. This is not only the first evolved VgrG described in Salmonella, but also the first evolved VgrG including a pyocin domain described so far in the literature. In addition, the data indicate that SPI-6 T6SS is widely distributed in S. enterica and absent in serotypes Enteritidis, Gallinarum, Agona, Javiana, Paratyphi B, Virchow, IIIa 62:z4,z23:- and IIIb 61:1,v:1,5,(7. Interestingly, while some serotypes harbor multiple T6SS (Dublin, Weltvreden and IIIa 62:z4,z23:- others do not encode for any (Enteritidis, Paratyphi B, Javiana, Virchow and IIIb 61:1,v:1,5,(7. Comparative and phylogenetic analyses indicate that the 4 T6SS loci in Salmonella have a distinct evolutionary history. Finally, we
Blondel, Carlos J; Jiménez, Juan C; Contreras, Inés; Santiviago, Carlos A
The recently described Type VI Secretion System (T6SS) represents a new paradigm of protein secretion in bacteria. A number of bioinformatic studies have been conducted to identify T6SS gene clusters in the available bacterial genome sequences. According to these studies, Salmonella harbors a unique T6SS encoded in the Salmonella Pathogenicity Island 6 (SPI-6). Since these studies only considered few Salmonella genomes, the present work aimed to identify novel T6SS loci by in silico analysis of every genome sequence of Salmonella available. The analysis of sequencing data from 44 completed or in progress Salmonella genome projects allowed the identification of 3 novel T6SS loci. These clusters are located in differentially-distributed genomic islands we designated SPI-19, SPI-20 and SPI-21, respectively. SPI-19 was identified in a subset of S. enterica serotypes including Dublin, Weltevreden, Agona, Gallinarum and Enteritidis. In the later, an internal deletion eliminated most of the island. On the other hand, SPI-20 and SPI-21 were restricted to S. enterica subspecies arizonae (IIIa) serotype 62:z4,z23:-. Remarkably, SPI-21 encodes a VgrG protein containing a C-terminal extension similar to S-type pyocins of Pseudomonas aeruginosa. This is not only the first evolved VgrG described in Salmonella, but also the first evolved VgrG including a pyocin domain described so far in the literature. In addition, the data indicate that SPI-6 T6SS is widely distributed in S. enterica and absent in serotypes Enteritidis, Gallinarum, Agona, Javiana, Paratyphi B, Virchow, IIIa 62:z4,z23:- and IIIb 61:1,v:1,5,(7). Interestingly, while some serotypes harbor multiple T6SS (Dublin, Weltvreden and IIIa 62:z4,z23:-) others do not encode for any (Enteritidis, Paratyphi B, Javiana, Virchow and IIIb 61:1,v:1,5,(7)). Comparative and phylogenetic analyses indicate that the 4 T6SS loci in Salmonella have a distinct evolutionary history. Finally, we identified an orphan Hcp-like protein
Klebsiella pneumoniae is a well known human pathogen. Although infectious in most nosocomial infections with a high level of resistance, capsular types and circulating hypervirulent strains in our context are not documented. The aims of this study are to identify capsular serotypes and hypervirulent strains circulating at the ...
The molecular serotypes and the evolutionary lineage of Listeria monocytogenes isolated from various foods in Nigeria are yet to be documented. Consequently, popular uncooked food items known locally as Okazi Utazi, Onugbu, Ogbono, Garri and Egusi obtained from plants botanically known as Gnetum africanum, ...
Forshey, Brett M.; Morrison, Amy C.; Cruz, Cristhopher; Rocha, Claudio; Vilcarromero, Stalin; Guevara, Carolina; Camacho, Daria E.; Alava, Araceli; Madrid, César; Beingolea, Luis; Suarez, Víctor; Comach, Guillermo
In 2008, dengue virus serotype 4 (DENV-4) emerged in northeastern Peru, causing a large outbreak and displacing DENV-3, which had predominated for the previous 6 years. Phylogenetic analysis of 2008 and 2009 isolates support their inclusion into DENV-4 genotype II, forming a lineage distinct from strains that had previously circulated in the region. PMID:19891873
The presence of Salmonella and human pathogens in unpasteurized milk remains a public health hazard. The study reported the phenotypic and molecular characterization of Salmonella serotypes in cow raw milk, cheese and traditional yoghurt marketed for man's consumption in Nigeria. Isolation of Salmonella was done ...
Dr DADIE Thomas
Feb 17, 2014 ... The virulence, serotype and phylogenetic traits of diarrhoeagenic Escherichia coli were detected in 502 strains isolated during digestive infections. Molecular detection of the target virulence genes, rfb gene of operon O and phylogenetic grouping genes Chua, yjaA and TSPE4.C2 was performed.
The carriage of Streptococcus pneumoniae, serotypes, antimicrobial susceptibility patterns and disease development are poorly understood in Yei. Availability of affordable antibiotics over the counter, lack of laboratory infrastructure and high rates of penicillin resistance have the potential to aggravate rates of childhood ...
Christensen, J.P.; Brown, D.J.; Madsen, Mogens
A substantial increase in the prevalence of S. enterica serovar Tennessee was observed in broiler flocks in Denmark at the turn of the year 1994 and in the following months. Epidemiological data indicated that a single hatchery was involved in spreading of the infection. Molecular characterization...... of S. enterica serovar Tennessee isolates from Danish broilers (1992 to 1995), the suspected hatchery and strains from various other sources included for comparison was initiated in order to trace the source of infection of the broilers. In general, strains of S. enterica ser. Tennessee showed only....... Restriction enzyme analysis of the plasmid ensured that the plasmids from broilers and the hatchery were identical. By analysis of cleaning and disinfection procedures and by sampling of different control points in the hatchery it was shown that S. enterica ser. Tennessee had colonized areas of the hatchers...
Dr. Reid Harvey, a CDC epidemiologist, discusses Salmonella enterica Dublin in People. Created: 10/5/2017 by National Center for Emerging and Zoonotic Infectious Diseases (NCEZID). Date Released: 10/5/2017.
Samuel, M.D.; Goldberg, Diana R.; Shadduck, D.J.; Price, J.I.; Cooch, E.G.
Pharyngeal swabs were collected from 298 lesser snow geese (Chen caerulescens caerulescens) at Banks Island (Northwest Territories. Canada) in the summer of 1994. Pasteurella multocida serotype 1 was isolated from an adult male bird and P. multocida serotype 3 was isolated from an adult female goose. Pathogenicity of the serotype 1 isolate was confirmed by inoculation in Pekin ducks (Anas platyrhynchos). The serotype 3 isolate was non-pathogenic in Pekin ducks. This is the first documented isolation of pathogenic P. multocida serotype 1 from apparently healthy wild snow geese.
Mikalová, Lenka; Bosák, Juraj; Hříbková, Hana; Dědičová, Daniela; Benada, Oldřich; Šmarda, Jan; Šmajs, David
Forty strains of Salmonella enterica (S. enterica) subspecies salamae (II), arizonae (IIIa), diarizonae (IIIb), and houtenae (IV) were isolated from human or environmental samples and tested for bacteriophage production. Production of bacteriophages was observed in 15 S. enterica strains (37.5%) belonging to either the subspecies salamae (8 strains) or diarizonae (7 strains). Activity of phages was tested against 52 pathogenic S. enterica subsp. enterica isolates and showed that phages produced by subsp. salamae had broader activity against pathogenic salmonellae compared to phages from the subsp. diarizonae. All 15 phages were analyzed using PCR amplification of phage-specific regions and 9 different amplification profiles were identified. Five phages (SEN1, SEN4, SEN5, SEN22, and SEN34) were completely sequenced and classified as temperate phages. Phages SEN4 and SEN5 were genetically identical, thus representing a single phage type (i.e. SEN4/5). SEN1 and SEN4/5 fit into the group of P2-like phages, while the SEN22 phage showed sequence relatedness to P22-like phages. Interestingly, while phage SEN34 was genetically distantly related to Lambda-like phages (Siphoviridae), it had the morphology of the Myoviridae family. Based on sequence analysis and electron microscopy, phages SEN1 and SEN4/5 were members of the Myoviridae family and phage SEN22 belonged to the Podoviridae family.
Glawischnig, Walter; Lazar, Judit; Wallner, Alice; Kornschober, Christian
Salmonella enterica serovar Dublin is endemic in the cattle population in some areas of the Austrian province Tyrol, and each year single dairy farms have experienced clinical infections. To ascertain if Tyrolean red foxes ( Vulpes vulpes ) act as a reservoir for Salmonella spp., we tested hepatic tissue and intestinal content from foxes hunted in the years 2015-16 by using microbiological methods. In addition, we included several fox fecal samples collected on a mountain pasture near chamois carcasses in the investigation. Of 434 foxes tested, nine animals (2.1%) were positive for Salmonella spp. Serotyping revealed five foxes positive with S. Dublin, demonstrating that this serovar exists in the Tyrolean fox population. The fecal samples collected in the area surrounding skeletonized chamois ( Rupicapra rupicapra ) also tested positive for S. Dublin. These chamois were probably victims of a waterborne outbreak caused by S. Dublin-shedding cattle. Our results indicate that the S. Dublin infections in red foxes were primarily acquired through ingestion of infected cattle material such as abortion tissues, but also by feeding on dead chamois. The findings underline the importance of interspecies transmission in this domestic/wildlife interface.
Rodríguez, Edna Catering; Díaz-Guevara, Paula; Moreno, Jaime; Bautista, Adriana; Montaño, Lucy; Realpe, María Elena; Della Gaspera, Anabella; Wiesner, Magdalena
Salmonella is an enteropathogen acquired through contaminated food or water. In Colombia, Salmonella spp. is included in the national surveillance of Acute Diarrhoeal Diseases and typhoid fever initiated in 1997. This report shows the phenotype and genotype results obtained from 2005 to 2011. A total of 4010 isolates of Salmonella enterica were analysed by serotyping with Kauffmann-White-LeMinor, antimicrobial resistance patterns, and pulse-field gel electrophoresis (PFGE). A total of 93 serovars were identified, of which, Typhimurium, Enteritidis, Typhi, Dublin, Panama, Derby, Braenderup, Saintpaul, and Uganda were prominent. The highest levels of resistance were found for tetracycline and nalidixic acid. Susceptibility was observed in 52.4% (2101/4010) of the isolates. Multi-resistance was recorded in 54.9% of Typhimurium isolates, with 81 different combinations. Using PFGE, 51.9% (2083/4010) isolates were analysed in 34 serovars, and 828 electrophoretic patterns were obtained. From these, 8 patterns were found in at least two Latin-American countries. The surveillance of Salmonella spp. provides information on the serovar distribution, antimicrobial resistance, and clonal distribution in Colombia, as well as information to treat this disease and control the spread of antimicrobial bacterial resistance. Copyright © 2016 Elsevier España, S.L.U. and Sociedad Española de Enfermedades Infecciosas y Microbiología Clínica. All rights reserved.
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.
Šplíchal, Igor; Trebichavský, Ilja; Šplíchalová, Alla; Barrow, P. A.
Roč. 103, - (2005), s. 155-161 ISSN 0165-2427 R&D Projects: GA ČR GA524/01/0917; GA MŠk ME 580 Institutional research plan: CEZ:AV0Z5020903 Keywords : salmonella typhymurium * gnotobiotic pig * cytokine Subject RIV: EE - Microbiology, Virology Impact factor: 1.626, year: 2005
Šplíchal, Igor; Trebichavský, Ilja; Šplíchalová, Alla; Barrow, P. A.
Roč. 103, - (2005), s. 155-161 ISSN 0165-2427 R&D Projects: GA ČR GA524/01/0917 Institutional research plan: CEZ:AV0Z5020903 Keywords : salmonella typhimurium * gnotobiotic pig * cytokine Subject RIV: EE - Microbiology, Virology Impact factor: 1.626, year: 2005
Pearse Lisa A
Full Text Available Abstract Background In December 2001, a fatal case of pneumococcal meningitis in a Marine Corps recruit was identified. As pneumococcal vaccine usage in recruit populations is being considered, an investigation was initiated into the causative serotype. Case presentation Traditional and molecular methods were utilized to determine the serotype of the infecting pneumococcus. The pneumococcal isolate was identified as serotype 38 (PS38, a serotype not covered by current vaccine formulations. The global significance of this serotype was explored in the medical literature, and found to be a rare but recognized cause of carriage and invasive disease. Conclusion The potential of PS38 to cause severe disease is documented in this report. Current literature does not support the hypothesis that this serotype is increasing in incidence. However, as we monitor the changing epidemiology of pneumococcal illness in the US in this conjugate era, PS38 might find a more prominent and concerning niche as a replacement serotype.
Wheeler, Emily; Cann, Isaac K O; Mackie, Roderick I
Salmonella carriage patterns in wild and captive reptiles suggest that both geographical proximity and host ecological differences may determine bacterial diversity among reptile populations. In this study, we explore the relative importance of these factors on Salmonella diversity in free-living Galápagos iguanas. We isolated Salmonella enterica from marine iguanas (Amblyrhynchus cristatus) and land iguanas (Conolophus subcristatus and C. pallidus) living on two islands (Plaza Sur and Santa Fe). We evaluated Salmonella population patterns using genomic fingerprints, sequence typing and serotyping. Rep-PCR fingerprinting revealed significant grouping of isolates by iguana population. Island residence had the strongest effect on isolate similarity, but a smaller divergence among Salmonella isolates from different iguana ecotypes (land versus marine) was detected within each island. In contrast, sequence typing detected a marginal difference in isolate genotypes between islands. Sequence types corresponded strongly to serotype identity, with both islands hosting a unique serovar pool. Our findings suggest that both geographical location and host ecotype differences (either from within host strain selection or from differences in habitat use) contribute to Salmonella population patterns in the Galápagos Islands. © 2010 Society for Applied Microbiology and Blackwell Publishing Ltd.
Bonardi, Silvia; Bassi, Luca; Brindani, Franco; D'Incau, Mario; Barco, Lisa; Carra, Elena; Pongolini, Stefano
In 2005-2008, 1152 samples (451 faecal samples, 451 carcass swabs and 250 tonsils) were collected from 451 finishing pigs slaughtered in three abattoirs of northern Italy. In two abattoirs, 34 scalding water samples were collected. The aim of this study was to investigate the faecal and palatine tonsil carriage rate of Salmonella enterica and Yersinia enterocolitica in pigs at slaughter and the degree of carcass contamination by these bacteria. Typing of the isolates, virulence characterization and antimicrobial testing were also performed. S. enterica was isolated from 21.5% of the faecal samples, 10.9% of the carcasses and 10.4% of the tonsils, but not from scalding water. Nineteen different serovars were identified among 172 S. enterica isolates. The prevalent serovars were Derby (41.3%), Rissen (12.2%), Typhimurium (11%), 4,,12:i:- (8.7%) and Give (4.1%). S. enterica ser. Typhimurium and S. enterica ser. 4,,12:i:- isolates were phage-typed and PT DT120 was the most common (23.5%). Y. enterocolitica was detected in 17.1% of the faecal samples, 2.4% of the carcasses, 10.8% of the tonsils and 11.8% of the scalding water samples. A total of 119 isolates were found, four of them in water. Of the 115 Y. enterocolitica isolates of pig origin, 24 (20.9%) were 4/O:3 and 4 (3.5%) were 2/O:9. Y. enterocolitica 4/O:3 represented 85.7% of the pathogenic isolates found in all types of samples and 100% of those found in tonsils. In 4/O:3 isolates the most common virulence-associated genes were ystA (100%), inv (95.8%), ail (87.5%) and yadA (54.2%). In 2/O:9 isolates the prevalent genes were ail (100%), inv (100%) and ystA (100%), followed by ystB (25.0%). The majority (75.7%) of Y. enterocolitica isolates was biotype 1A, belonging to 13 serotypes (O:3; O:5; O:4,32-4,33; O:6,30-6,31; O:7,8-8; O:7,8-8-8,19; O:7,13; O:8; O:9; O:13; O:16-16,29; O:41,42-41,43; O:52). The most common virulence genes in 1A isolates were inv (95.4%) and ystB (72.4%). The antimicrobial
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 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.
SACCHI Claudio Tavares
Full Text Available In the present study we examine the potential use of oligonucleotide probes to characterize Neisseria meningitidis serotypes without the use of monoclonal antibodies (MAbs. Antigenic diversity on PorB protein forms the bases of serotyping method. However, the current panel of MAbs underestimated, by at least 50% the PorB variability, presumably because reagents for several PorB variable regions (VRs are lacking, or because a number of VR variants are not recognized by serotype-defining MAbs12. We analyzed the use of oligonucleotide probes to characterize serotype 10 and serotype 19 of N. meningitidis. The porB gene sequence for the prototype strain of serotype 10 was determined, aligned with 7 other porB sequences from different serotypes, and analysis of individual VRs were performed. The results of DNA probes 21U (VR1-A and 615U (VR3-B used against 72 N. meningitidis strains confirm that VR1 type A and VR3 type B encode epitopes for serotype-defined MAbs 19 and 10, respectively. The use of probes for characterizing serotypes possible can type 100% of the PorB VR diversity. It is a simple and rapid method specially useful for analysis of large number of samples.
Hwang, Jeong-Hwan; Shin, Gee-Wook; Hwang, Joo-Hee; Lee, Chang-Seop
A 57-year-old man presented with high fever and diarrhea. A blood culture revealed the presence of a Group C nontyphoidal Salmonella (NTS) isolate. On Salmonella serotyping, the isolate was identified as Salmonella enterica serovar Virchow. Its sequence type was determined to be ST16 by sequence analysis of 7 different housekeeping genes. The bla CTX-M group 1 and bla TEM genes were amplified using multiplex PCR assay for detecting extended-spectrum β-lactamases (ESBL) genes. Sequences of both amplicons were respectively identical to CTX-M-15- and TEM-1-encoding genes. Since NTS is a cause of foodborne illness outbreaks in communities and an important cause of community-acquired bloodstream infection, clinicians should consider ESBL- or AmpC-producing NTS species in the differential diagnosis.
Full Text Available Salmonella bacteria are among the most frequent causes of food poisoning, and milk and milk products are food items through which these bacteria can be transmitted. More recent data indicate that the infective dose of Salmonella is very small - up to 101/g/ml food. The preservation of Salmonella in milk depends on the acidity, temperature and the duration of storage. The objective of the work was to examine the survival of Salmonella in milk depending on the changes of the total number of bacteria, pH and temperature. In the study, 0.1 ml of 24-hour bouillon culture of S. enteritidis was inoculated in raw collective milk (104 cfu/ml and in marketed pasteurized milk (102 cfu/ml each. The examined milk samples were maintained at room temperature and at a temperature of 7oC. At the beginning of the examinations, after 24h, 48h, 72h, after 7, 14 and 21 days, decimal solutions were made from the milk, and, following incubation, the number of S. enteritidis was determined (Rambach agar, the total number of bacteria (base for total number of bacteria by multiplying the number of grown colonies and the corresponding solution, and the milk pH was measured. There was a bigger reduction of S. enteritidis at room temperature because the pH value dropped more swiftly. The lowest pH at which S. enteritidis was isolated from experimentally contaminated milk was 3.88. The results of the examinations show that Salmonella can adapt to biochemical processes during milk acid fermentation of soft cheeses. No Salmonella bacteria were isolated from experimentally contaminated pasterurized milk at both maintenance temperatures after seven days of investigations.
Manesh, Abi; Balaji, Veeraraghavan; Kumar, Devanga Ragupathi Naveen; Rupali, Priscilla
Typhoid fever remains a serious problem in many developing countries. Due to resistance to multiple first line drugs, azithromycin has evolved as an important drug in the treatment of typhoid. While therapy with azithromycin is highly effective, no clinically validated mean inhibitory concentration (MIC) break points or disc diffusion cutoff guidelines are available so far. We describe an Indian adult with clinical and microbiological failure to azithromycin despite low azithromycin MIC. Copyright © 2016. Published by Elsevier Ltd.
html . 4. Olson AB, Andrysiak AK, Tracz DM, Guard-Bouldin J, Demczuk W, Ng LK, Maki A, Jamieson F, Gilmour MW. 2007. Limited genetic diversity in...06. 33. Hyeon JY, Chon JW, Park JH, Kim MS, Oh YH, Choi IS, Seo KH. 2013. A comparison of subtyping methods for differentiating Salmonella en- terica
Andino, A.; Hanning, I.
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
Hoffmann, Maria; Pettengill, James B; Gonzalez-Escalona, Narjol; Miller, John; Ayers, Sherry L; Zhao, Shaohua; Allard, Marc W; McDermott, Patrick F; Brown, Eric W; Monday, Steven R
Determinants of multidrug resistance (MDR) are often encoded on mobile elements, such as plasmids, transposons, and integrons, which have the potential to transfer among foodborne pathogens, as well as to other virulent pathogens, increasing the threats these traits pose to human and veterinary health. Our understanding of MDR among Salmonella has been limited by the lack of closed plasmid genomes for comparisons across resistance phenotypes, due to difficulties in effectively separating the DNA of these high-molecular weight, low-copy-number plasmids from chromosomal DNA. To resolve this problem, we demonstrate an efficient protocol for isolating, sequencing and closing IncA/C plasmids from Salmonella sp. using single molecule real-time sequencing on a Pacific Biosciences (Pacbio) RS II Sequencer. We obtained six Salmonella enterica isolates from poultry, representing six different serovars, each exhibiting the MDR-Ampc resistance profile. Salmonella plasmids were obtained using a modified mini preparation and transformed with Escherichia coli DH10Br. A Qiagen Large-Construct kit™ was used to recover highly concentrated and purified plasmid DNA that was sequenced using PacBio technology. These six closed IncA/C plasmids ranged in size from 104 to 191 kb and shared a stable, conserved backbone containing 98 core genes, with only six differences among those core genes. The plasmids encoded a number of antimicrobial resistance genes, including those for quaternary ammonium compounds and mercury. We then compared our six IncA/C plasmid sequences: first with 14 IncA/C plasmids derived from S. enterica available at the National Center for Biotechnology Information (NCBI), and then with an additional 38 IncA/C plasmids derived from different taxa. These comparisons allowed us to build an evolutionary picture of how antimicrobial resistance may be mediated by this common plasmid backbone. Our project provides detailed genetic information about resistance genes in
Allard, Sarah; Reynolds, Sara; Millner, Patricia; Arce, Gabriela; Blodgett, Robert J.; Brown, Eric W.
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
Full Text Available Abstract Background Shigella flexneri is the major pathogen causing bacillary dysentery. Fifteen serotypes have been recognized up to now. The genesis of new S. flexneri serotypes is commonly mediated by serotype-converting bacteriophages. Untypeable or novel serotypes from natural infections had been reported worldwide but have not been generated in laboratory. Results A new S. flexneri serotype-serotype 1 d was generated when a S. flexneri serotype Y strain (native LPS was sequentially infected with 2 serotype-converting bacteriophages, SfX first and then SfI. The new serotype 1 d strain agglutinated with both serotype X-specific anti-7;8 grouping serum and serotype 1a-specific anti- I typing serum, and differed from subserotypes 1a, 1b and 1c. Twenty four S. flexneri clinical isolates of serotype X were all converted to serotype 1 d by infection with phage SfI. PCR and sequencing revealed that SfI and SfX were integrated in tandem into the proA-yaiC region of the host chromosome. Conclusions These findings suggest a new S. flexneri serotype could be created in nature. Such a conversion may be constrained by susceptibility of a strain to infection by a given serotype-converting bacteriophage. This finding has significant implications in the emergence of new S. flexneri serotypes in nature.
Rochat, Tatiana; Fujiwara-Nagata, Erina; Calvez, Ségolène
Flavobacterium psychrophilum is a devastating bacterial pathogen of salmonids reared in freshwater worldwide. So far, serological diversity between isolates has been described but the underlying molecular factors remain unknown. By combining complete genome sequence analysis and the serotyping me...... for bacterial coldwater disease resistance and future vaccine formulation....
Potnis, Neha; Colee, James; Jones, Jeffrey B.
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
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.
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
The prevalence of Shigella serotypes and their sensitivity pattern was studied from January 2000 to December 2002 for a period of 3 years. Of the 2420 pediatric diarrhoeal stools screened, 84 Shigella organisms were isolated giving an isolation rate of 3.5%. S.flexneri was the predominant serotype (64%) followed by ...
Full Text Available Objective: Streptococcus agalactiae (Group B streptococci, GBS are frequently responsible for sepsis and meningitis seen in the early weeks of life. GBS may cause perinatal infection and premature birth in pregnant women. The aim of this study was to serotype GBS strains isolated from clinical samples and evaluate their serotype distribution according to their susceptibilities to antibiotics and isolation sites. Material and Methods: One hundred thirty one S. agalactiae strains isolated from the clinical samples were included in the study. Of the strains, 99 were isolated from urine, 20 from soft tissue, 10 from blood and 2 from vaginal swab. Penicillin G and ceftriaxone susceptibilities of GBS were determined by the agar dilution method. Susceptibilities to erythromycin, clindamycin, vancomycin and tetracycline were determined by the Kirby-Bauer method according to CLSI criteria. Serotyping was performed using the latex aglutination method using specific antisera (Ia, Ib, II-VIII. Results: While in 131 GBS strains, serotypes VII and VIII were not detected, the most frequently isolated serotypes were types Ia (36%, III (30.5% and II (13% respectively. Serotype Ia was the most frequently seen serotype in all samples. All GBS isolates were susceptible to penicilin G, ceftriaxone and vancomycin. Among the strains, tetracycline, erythromycin and clindamycin resistance rates were determined as 90%, 14.5%, and 13% respectively. Conclusion: Penicillin is still the first choice of treatment for the infections with all serotypes of S. agalactiae in Turkey.
Gates-Hollingsworth, Marcellene A; Kozel, Thomas R
To meet the needs of a global community, an immunoassay for cryptococcal antigen (CrAg) must have high sensitivity for CrAg of all major serotypes. A new immunoassay for CrAg in lateral flow format was evaluated and found to have a high sensitivity for detection of serotypes A, B, C, and D.
Sep 15, 2009 ... A recombinant His-Vi protein of Salmonella enterica serovar Typhi was successfully constructed and cloned into ... mainly through consumption of food or water contami- nated with .... and healthy individuals (double arrows) followed by the detection using recombinant His-Vi protein as the primary antibody ...
Apr 17, 2013 ... food borne diseases in England and Wales (Adak et al.,. 2002). In the USA, about one million ... as a source of human foodborne diseases (Humphrey,. 2000; Adak et al., 2005). Apart from the .... rumen protozoan-mediated enhancement of invasion for multiple- antibiotic-resistant Salmonella enterica ...
Agron, Peter G. (Castro Valley, CA); Andersen, Gary L. (Berkeley, CA); Walker, Richard L. (Davis, CA)
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.
The susceptibility of 41 Salmonella enterica isolates isolated from poultry disease outbreaks between April 2005 and August 2007 in Oyo and Ogun States Nigeria to ciprofloxacin, nalidixic acid, chloramphenicol, streptomycin, kanamycin, ampicil in, neomycin, and tetracycline was determined. Using microtitre method for ...
Blehert, David S.; Hernandez, Sonia M.; Keel, Kevin; Sanchez, Susan; Trees, Eija; ,
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.
Serovariedades de Salmonella enterica subespecie enterica en porcinos de faena y su resistencia a los antimicrobianos Serovars of Salmonella enterica subspecies enterica and its antimicrobial resistance in slaughterhouse pigs
M. P. Ibar
Full Text Available Se realizó un estudio para determinar la prevalencia de Salmonella y sus serovariedades en cerdos de faena, para evaluar sus perfiles de resistencia a los antimicrobianos y para conocer la presencia de integrones de clase 1 como posibles reservorios de resistencia. A partir de un total de 386 muestras de porcinos provenientes de cuatro frigoríficos de las provincias de Buenos Aires y de Santa Fe (Argentina, se identificaron 93 (24,1% cepas de Salmonella enterica subespecie enterica, 52 (55,9% de contenido cecal y 41 (44,1% de nódulo linfático ileocecal. Se hallaron 13 serovariedades de S. enterica, las más prevalentes fueron S. Schwarzengrund, S. Heidelberg, S. subespecie I 6,8:e,h:-, S. Derby y S. Bredeney. Se probaron 15 antimicrobianos por el método de dilución en agar: amikacina, gentamicina, ciprofloxacina, cefalotina, cefotaxima, enrofloxacina, fosfomicina, polimixina-B, tetraciclina, cloranfenicol, estreptomicina, trimetoprima-sulfametoxazol, ampicilina, nitrofurantoína y ácido nalidíxico. Según se estableció mediante la determinación de la CIM, el 73% de las cepas de S. enterica subespecie enterica fueron sensibles a todos los antimicrobianos probados. Se observó resistencia a tetraciclina en 24 (25,8% de las 93 cepas, a cloranfenicol en 22 (23,7%, a estreptomicina en 22 (23,7% a trimetoprima-sulfametoxazol en 20 (21,5%, a ampicilina en 18 (19,4%, a nitrofurantoína en 3 (3,2% y a ácido nalidíxico en 3 (3,2%. Algunos aislamientos de S. Typhimurium, S. Heildelberg, S. Derby y S. Orion presentaron multirresistencia y portaban el gen de la integrasa clase 1. Los mayores porcentajes de resistencia correspondieron a los antimicrobianos habitualmente utilizados en veterinaria y en las explotaciones porcinas.A study was carried out in order to determine the prevalence of Salmonella and its serovars among porcine slaughterhouses, to evaluate the antimicrobial resistance profiles and to know the presence of class 1 integrons as
Gu, Ganyu; Hu, Jiahuai; Cevallos-Cevallos, Juan M.; Richardson, Susanna M.; Bartz, Jerry A.; van Bruggen, Ariena H. C.
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 (Penterica 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
Turner, Paul; Hinds, Jason; Turner, Claudia; Jankhot, Auscharee; Gould, Katherine; Bentley, Stephen D.; Nosten, François; Goldblatt, David
Identification of Streptococcus pneumoniae in the nasopharynx is critical for an understanding of transmission, estimates of vaccine efficacy, and possible replacement disease. Conventional nasopharyngeal swab (NPS) culture and serotyping (the WHO protocol) is likely to underestimate multiple-serotype carriage. We compared the WHO protocol with methods aimed at improving cocolonization detection. One hundred twenty-five NPSs from an infant pneumococcal-carriage study, containing ≥1 serotype by WHO culture, were recultured in duplicate. A sweep of colonies from one plate culture was serotyped by latex agglutination. DNA extracted from the second plate was analyzed by S. pneumoniae molecular-serotyping microarray. Multiple serotypes were detected in 11.2% of the swabs by WHO culture, 43.2% by sweep serotyping, and 48.8% by microarray. Sweep and microarray were more likely to detect multiple serotypes than WHO culture (P microarray and sweep, but the microarray identified the greatest number of serotypes. A common serogroup type was identified in 95.2% of swabs by all methods. WHO methodology significantly underestimates multiple-serotype carriage compared to these alternate methods. Sweep serotyping is cost-effective and field deployable but may fail to detect serotypes at low abundance, whereas microarray serotyping is more costly and technology dependent but may detect these additional minor carried serotypes. PMID:21411589
Full Text Available WHO estimated 50 million dengue infections happen every year in the world. In Indonesia, there were 90,245 DHF cases on 2012 with 816 deaths. In the Province of Aceh, 2,269 cases happened in the same year. This study aimed to identify dengue virus serotype in Aceh. Sampling was done in Kota Banda Aceh Hospital, Kota Lhokseumawe Hospital, Kabupaten Aceh Tamiang Hospital, Kabupaten Aceh Barat Hospital, and Kabupaten Simeulue Hospital between May to December 2012. This was a clinical laboratory research with observation design using cross sectional approach. Research’s population was sample from patients with dengue clinical symptom. Using purposive sampling technique, we have collected 100 samples from the five hospitals (20 samples from each hospital. From RT-PCR, we found 16 positive samples (9 samples were DENV-4, 3 samples were DENV-1, 2 samples were DENV-2, and 2 samples were DENV-3.
Nicholas J Croucher
Full Text Available 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.
Noriega, F R; Liao, F M; Maneval, D R; Ren, S; Formal, S B; Levine, M M
Based upon the lipopolysaccharide (LPS) structure and antigenicity of Shigella group B, a strategy for broad cross-protection against 14 Shigella flexneri serotypes was designed. This strategy involves the use of two S. flexneri serotypes (2a and 3a), which together bear the all of the major antigenic group factors of this group. The novel attenuated strains used in these studies were S. flexneri 2a strain CVD 1207 (DeltaguaB-A DeltavirG Deltaset1 Deltasen) and S. flexneri 3a strain CVD 1211 (DeltaguaB-A DeltavirG Deltasen). Guinea pigs were immunized with an equal mixture of these strains and later challenged (Sereny test) with a wild-type S. flexneri serotype 1a, 1b, 2b, 4b, 5b, Y, or 6 strain of demonstrated virulence in the same model. Guinea pigs that were immunized with these two vaccine strains produced serum and mucosal antibodies that cross-reacted with all the S. flexneri serotypes tested (except of S. flexneri serotype 6) as assessed by enzyme-linked immunosorbent assay, immunoblotting, and slide agglutination. Furthermore, the combination vaccine conferred significant protection against challenge with S. flexneri serotypes 1b, 2b, 5b, and Y but not with serotypes 1a, 4b, or (as predicted) 6.
Brian L Pike
Full Text Available 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.
Jean-Gilles Beaubrun, J; Tall, Ben D; Flamer, M-L; Patel, I; Gopinath, G; Auguste, Winny; Jean, Catherine; George, Melvin; Tartera, Carmen; Ewing, L; Hanes, D E
During an investigation to increase the recovery of Salmonella enterica from Oregano, an increased expression of exopolysaccharide was induced in Salmonella serovar Montevideo. The atypical mucoid (SAL242S) and the non-mucoid (SAL242) strains of Montevideo were compared and characterized using various methods. Serotyping analysis demonstrated that both strains are the same serovar Montevideo. Electron microscopy (EM) of cultured SAL242S cells revealed the production of a prominent EPS-like structure enveloping aggregates of cells that are composed of cellulose. Mucoid cells possessed a higher binding affinity for Calcofluor than that of the non-mucoid strain. Genotypic analysis revealed no major genomic differences between these morphotypes, while expression analyses using a DNA microarray shows that the mucoid variant exhibited heightened expression of genes encoding proteins produced by the SPI-1 type III secretion system. This increased expression of SPI1 genes may play a role in protecting Salmonella from environmental stressors. Based on these observations, Salmonella serovar Montevideo mucoid variant under stressful or low-nutrient environments presented atypical growth patterns and phenotypic changes, as well as an upregulated expression of virulence factors. These findings are significant in the understanding of survival abilities of Salmonella in a various food matrices. Published by Elsevier Ltd.
Cooley, Michael B; Quiñones, Beatriz; Oryang, David; Mandrell, Robert E; Gorski, Lisa
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.
Ives, A-K; Antaki, E; Stewart, K; Francis, S; Jay-Russell, M T; Sithole, F; Kearney, M T; Griffin, M J; Soto, E
Salmonellae are Gram-negative zoonotic bacteria that are frequently part of the normal reptilian gastrointestinal flora. The main objective of this project was to estimate the prevalence of non-typhoidal Salmonella enterica in the nesting and foraging populations of sea turtles on St. Kitts and in sand from known nesting beaches. Results suggest a higher prevalence of Salmonella in nesting leatherback sea turtles compared with foraging green and hawksbill sea turtles. Salmonella was cultured from 2/9 and identified by molecular diagnostic methods in 3/9 leatherback sea turtle samples. Salmonella DNA was detected in one hawksbill turtle, but viable isolates were not recovered from any hawksbill sea turtles. No Salmonella was detected in green sea turtles. In samples collected from nesting beaches, Salmonella was only recovered from a single dry sand sample. All recovered isolates were positive for the wzx gene, consistent with the O:7 serogroup. Further serotyping characterized serovars Montevideo and Newport present in cloacal and sand samples. Repetitive-element palindromic PCR (rep-PCR) fingerprint analysis and pulsed-field gel electrophoresis of the 2014 isolates from turtles and sand as well as archived Salmonella isolates recovered from leatherback sea turtles in 2012 and 2013, identified two distinct genotypes and four different pulsotypes, respectively. The genotyping and serotyping were directly correlated. To determine the persistence of representative strains of each serotype/genotype in these environments, laboratory-controlled microcosm studies were performed in water and sand (dry and wet) incubated at 25 or 35°C. Isolates persisted for at least 32 days in most microcosms, although there were significant decreases in culturable bacteria in several microcosms, with the greatest reduction in dry sand incubated at 35°C. This information provides a better understanding of the epizootiology of Salmonella in free-ranging marine reptiles and the potential
Angen, Øystein; Heegaard, Peter M. H.; Lavritsen, D.T.
In Denmark porcine pleuropneumonia is most frequently caused by Actinobacillus pleuropneumoniae serotype 2 (60%). Isolation of A. pleuropneumoniae from nasal cavities or tonsils from carrier animals is complicated due to the mixed bacterial flora present. An immunomagnetic separation technique (I...
L. Katzelnick (Leah); J.M. Fonville (Judith); G.D. Gromowski (Gregory D.); J.B. Arriaga (Jose Bustos); A. Green (Angela); S.L. James (Sarah ); L. Lau (Louis); M. Montoya (Magelda); C. Wang (Chunling); L.A. Van Blargan (Laura A.); C.A. Russell (Colin); H.M. Thu (Hlaing Myat); T.C. Pierson (Theodore C.); P. Buchy (Philippe); J.G. Aaskov (John G.); J.L. Muñoz-Jordán (Jorge L.); N. Vasilakis (Nikos); R.V. Gibbons (Robert V.); R.B. Tesh (Robert B.); A.D.M.E. Osterhaus (Albert); R.A.M. Fouchier (Ron); A. Durbin (Anna); C.P. Simmons (Cameron P.); E.C. Holmes (Edward C.); E. Harris (Eva); S.S. Whitehead (Stephen S.); D.J. Smith (Derek James)
textabstractThe four genetically divergent dengue virus (DENV) types are traditionally classified as serotypes. Antigenic and genetic differences among the DENV types influence disease outcome, vaccine-induced protection, epidemic magnitude, and viral evolution.We scharacterized antigenic diversity
Hoorfar, Jeffrey; Ahrens, Peter; Rådström, P.
A simple and ready-to-go test based on a 5' nuclease (TaqMan) PCR technique was developed for identification of presumptive Salmonella enterica isolates. The results were compared with those of conventional methods. The TaqMan assay was evaluated for its ability to accurately detect 210 S. enterica...... isolates, including 100 problematic "rough" isolates. An internal positive control was designed to use the same Salmonella primers for amplification of a spiked nonrelevant template (116 bp) in the sample tube. The PCR test correctly identified all the Salmonella strains by resulting in positive end......-point fluorescence (FAM) signals for the samples and positive control (TET) signals (relative sensitivity [Delta Rn], >0.6). The diagnostic specificity of the method was assessed using 120 non-Salmonella strains, which all resulted in negative FAM signals (Delta Rn, less than or equal to 0.5). All 100 rough...
Martens, Pernille; Worm, Signe Westring; Lundgren, Bettina
Serotype-specific mortality from invasive Streptococcus pneumoniae disease revisited.Martens P, Worm SW, Lundgren B, Konradsen HB, Benfield T. Department of Infectious Diseases 144, Hvidovre University Hospital, DK-2650 Hvidovre, Denmark. firstname.lastname@example.org BACKGROUND: Invasive infection w...... pneumococcal disease. The limitations of the current polysaccharide pneumococcal vaccine warrant the development of alternative vaccines. We suggest that the virulence of pneumococcal serotypes should be considered in the design of novel vaccines....
Full Text Available The methylation of DNA bases plays an important role in numerous biological processes including development, gene expression, and DNA replication. Salmonella is an important foodborne pathogen, and methylation in Salmonella is implicated in virulence. Using single molecule real-time (SMRT DNA-sequencing, we sequenced and assembled the complete genomes of eleven Salmonella enterica isolates from nine different serovars, and analysed the whole-genome methylation patterns of each genome. We describe 16 distinct N6-methyladenine (m6A methylated motifs, one N4-methylcytosine (m4C motif, and one combined m6A-m4C motif. Eight of these motifs are novel, i.e., they have not been previously described. We also identified the methyltransferases (MTases associated with 13 of the motifs. Some motifs are conserved across all Salmonella serovars tested, while others were found only in a subset of serovars. Eight of the nine serovars contained a unique methylated motif that was not found in any other serovar (most of these motifs were part of Type I restriction modification systems, indicating the high diversity of methylation patterns present in Salmonella.
Aksoy, Deniz; Şen, Ece
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
Kramer, B; Wunderlich, J; Muranyi, P
The objective of this study was a comprehensive characterization of physiological changes of Salmonella enterica induced by intense broad spectrum pulsed light (PL). After exposing the bacteria to this nonthermal decontamination technology on a gel surface, multiple viability parameters beyond culturability were assessed. By applying flow cytometry, a luciferin-luciferase bioluminescence assay and a microplate assay to measure the current redox activity, the impact of pulsed light on the membrane potential, membrane integrity, esterase activity, efflux pump activity, expression of the green fluorescent protein (GFP), respiration activity and ATP-content of Salm. enterica ATCC BAA-1045 was determined. These culture-independent methods for assessing the bacterial activity were compared to the ability to grow on tryptic soy agar. It is shown that this strain is rather sensitive to PL considering colony count reductions, while on the other hand unculturable bacteria still exhibit significant cellular energetic functions. However, this residual activity after PL exposure significantly decreases during sample storage in buffer for 24 h. This study also shows that the GFP expression of PL-treated cells which have rendered unculturable is severely reduced. This study reveals that although not all cellular functions of Salm. enterica are immediately shut down after PL exposure, the synthesis of new GFP is strongly reduced and affected to a similar extent as the culturability. It is shown for the first time, that even there is significant bacterial activity measurable after PL exposure, it is likely that nongrowing pathogenic bacteria like Salm. enterica are unable to express proteins, which is of great importance regarding their pathogenicity. © 2016 The Society for Applied Microbiology.
Jeri D Barak
Full Text Available 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 of direct artificial inoculation have not been investigated. METHODOLOGY/PRINCIPAL FINDINGS: This work examined the role of contaminated soil, the potential for crop debris to act as inoculum from one crop to the next, and any interaction between the seedbourne plant pathogen Xanthomonas campestris pv. vesicatoria and S. enterica on tomato plants. Our results show S. enterica can survive for up to six weeks in fallow soil with the ability to contaminate tomato plants. We found S. enterica can contaminate a subsequent crop via crop debris; however a fallow period between crop incorporation and subsequent seeding can affect contamination patterns. Throughout these studies, populations of S. enterica declined over time and there was no bacterial growth in either the phyllosphere or rhizoplane. The presence of X. campestris pv. vesicatoria on co-colonized tomato plants had no effect on the incidence of S. enterica tomato phyllosphere contamination. However, growth of S. enterica in the tomato phyllosphere occurred on co-colonized plants in the absence of plant disease. CONCLUSIONS/SIGNIFICANCE: S. enterica contaminated soil can lead to contamination of the tomato phyllosphere. A six week lag period between soil contamination and tomato seeding did not deter subsequent crop contamination. In the absence of plant disease, presence of the bacterial plant pathogen, X. campestris pv. vesicatoria was beneficial to S. enterica allowing multiplication of the human pathogen population. Any event leading to soil contamination with S. enterica could pose a public health risk with subsequent tomato production, especially in areas prone to bacterial spot disease.
Zheng, Zhibei; Zheng, Wei; Wang, Haoqiu; Pan, Jincao; Pu, Xiaoying
Currently, no protocols or commercial kits are available to determine the serotypes of Salmonella by using Luminex MAGPIX®. In this study, an xTAG assay for serotype determination of Salmonella suitable for Luminex MAGPIX® is described and 228 Salmonella isolates were serotype determined by this xTAG assay. The xTAG assay consists of two steps: 1) Multiplex PCR to amplify simultaneously O, H and Vi antigen genes of Salmonella, and 2) Magplex-TAG™ microsphere hybridization to identify accurately the specific PCR products of different antigens. Compared with the serotyping results of traditional serum agglutination test, the sensitivity and specificity of the xTAG assay were 95.1% and 100%, respectively. The agreement rate of these two assays was 95.2%. Compared with Luminex xMAP® Salmonella Serotyping Assay (SSA) kit, the advantages of this xTAG assay are: First, the magnetic beads make it applicable to both the Luminex®100/200™ and MAGPIX® systems. Second, only primers rather than both primers and probes are needed in the xTAG assay, and the process of coupling antigen-specific oligonucleotide probes to beads is circumvented, which make the xTAG assay convenient to be utilized by other laboratories. The xTAG assay may serve as a rapid alternative or complementary method for traditional Salmonella serotyping tests, especially for laboratories that utilize the MAGPIX® systems. Copyright © 2017 Elsevier B.V. All rights reserved.
Bailey, J S; Cox, N A; Craven, S E; Cosby, D E
The widespread presence of Salmonella in all phases of broiler chicken production and processing is well documented. However, little information is available to indicate the identity and movement of specific serotypes of Salmonella through the different phases of an integrated operation. In this study, samples were collected from the breeder farm, from the hatchery, from the previous grow-out flock, from the flock during grow-out, and from carcasses after processing. Salmonella were recovered from 6, 98, 24, 60, and 7% of the samples, respectively, in the first trial and from 7, 98, 26, 22, and 36% of the samples, respectively, in the second trial. Seven different serotypes were identified in the first trial, and 12 different serotypes were identified in the second trial. For both trials there was poor correlation between the serotypes found in the breeder farms and those found in the hatchery. This finding and the fact that similar serotypes were found in the hatchery in both trials suggests that there was an endemic population of Salmonella in the hatchery. An association between the serotypes found in the hatchery and those found on the final processed carcasses was observed in both trials. This study confirms that a successful intervention program for broiler production operations must be multifaceted, with one component being disinfection in the hatchery.
Turner, Paul; Turner, Claudia; Jankhot, Auscharee; Phakaudom, Kawalee; Nosten, Francois; Goldblatt, David
Background Nasopharyngeal swab (NPS) culture by World Health Organisation (WHO) methodology underestimates multiple pneumococcal serotype colonisation compared to a simple culture and latex sweep method. The impacts of this on descriptions of pneumococcal serotype distributions and colonisation dynamics in infancy are not clear. Methods 8,736 NPS collected from infants enrolled into a longitudinal study were processed to evaluate the field utility of the latex sweep method. 1,107 had previously been cultured by WHO methodology. Additionally, colonisation results were compared in 100 matched pairs of infants, where swabs from an individual were cultured either by WHO or latex sweep method. Results In 1,107 swabs cultured by both methods, the latex sweep method was three times more likely to detect colonisation with multiple pneumococcal serotypes than the WHO method (platex sweep (p = 0.03). The overall rate of non-vaccine type pneumococcal acquisition was also greater in infants cultured by latex sweep (p = 0.04). Conclusions Latex sweep serotyping was feasible to perform on a large specimen collection. Multiple serotype colonisation detection was significantly improved compared with WHO methodology. However, use of the latex sweep method is unlikely to significantly alter colonisation study serotype distribution or colonisation dynamics results. PMID:23844133
Klerks, M.M.; Gent-Pelzer, van M.P.E.; Franz, E.; Zijlstra, C.; Bruggen, van A.H.C.
This paper describes the physiological and molecular interactions between the human-pathogenic organism Salmonella enterica serovar Dublin and the commercially available mini Roman lettuce cv. Tamburo. The association of S. enterica serovar Dublin with lettuce plants was first determined, which
Pedersen, Karl; Sørensen, Gitte; Szabo, Istvan
We report here the appearance of a very rare serovar of Salmonella, S. enterica subsp. enterica serovar Goverdhan, in routine Salmonella surveillance samples from Danish poultry production. S. Goverdhan was found on nine occasions: in one broiler breeder farm in October 2010, four broiler farms a...
Parkhill, J.; Dougan, G.; James, K.D.
Salmonella enterica serovar Typhi (S. typhi) is the aetiological agent of typhoid fever, a serious invasive bacterial disease of humans with an annual global burden of approximately 16 million cases, leading to 600,000 fatalities(1). Many S. enterica serovars actively invade the mucosal surface o...... plasmid of Yersinia pestis....
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...
Straaten, van T.; Janssen, R.; Mevius, D.J.; Dissel, van J.T.
MarA and its homologue, RamA, have been implicated in multidrug resistance (MDR). RamA overexpression in Salmonella enterica serovar Typhimurium and Escherichia coli conferred MDR independently of marA. Inactivation of ramA did not affect the antibiotic susceptibilities of wild-type S. enterica
Patterson, Sheila K; Kim, Hyeun Bum; Borewicz, Klaudyna; Isaacson, Richard E
Salmonella enterica is an important food borne pathogen that is frequently carried by swine. Carrier animals pose a food safety risk because they can transmit S. enterica to finished food products in the processing plant or by contamination of the environment. Environmental contamination has become increasingly important as non-animal foods (plant-based) have been implicated as sources of S. enterica. The prevalence of S. enterica in swine is high and yet carrier animals remain healthy. S. enterica has developed a highly sophisticated set of virulence factors that allow it to adapt to host environments and to cause disease. It is assumed that S. enterica also has developed unique ways to maintain itself in animals and yet not cause disease. Here we describe our research to understand persistence. Specifically, data are presented that demonstrates that detection of most carrier animals requires specific stresses that cause S. enterica to be shed from pigs. As well, we describe a phenotypic phase variation process that appears to be linked to the carrier state and a complex set of factors that control phenotypic phase variation. Finally, we describe how the composition of the gut bacterial microbiome may contribute to persistence and at the least how S. enterica might alter the composition of the gut bacterial microbiome.
Lee, Ki-Eun; Lee, Deog-Yong; Choi, Hwan-Won; Chae, Su-Jin; Yun, Young-Sun; Lee, Ki-Chan; Cho, Yun-Sang; Yang, Dong-Kun
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,,12:i:-. Salmonella 4,,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,,12:i:- strains, one (KVCC-BA1400078) was identified as a Salmonella 4,,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,,12:i:- in Korea poses a significant threat of horizontal transfer between pigs and humans.
Hoffmann, Maria; Payne, Justin; Roberts, Richard J; Allard, Marc W; Brown, Eric W; Pettengill, James B
Within the last several years, Salmonella enterica subsp. enterica serovar Agona has been among the 20 most frequently isolated serovars in clinical cases of salmonellosis. In this report, the complete genome sequence of S. Agona strain 460004 2-1 isolated from unsweetened puffed-rice cereal during a multistate outbreak in 2008 was sequenced using single-molecule real-time DNA sequencing. Copyright © 2015 Hoffmann et al.
Tomiyama, M P O; Werle, C H; Milanez, G P; Nóbrega, D B; Pereira, J P; Calarga, A P; Flores, F; Brocchi, M
Salmonella enterica subsp enterica serovar 4,5,12:i:- has been responsible for many recent Salmonella outbreaks worldwide. Several studies indicate that this serovar originated from S. enterica subsp enterica serovar Typhimurium, by the loss of the flagellar phase II gene (fljB) and adjacent sequences. However, at least two different clones of S. enterica 4,5,12:i:- exist that differs in the molecular events responsible for fljB deletion. The aim of this study was to test the stability of the fljBA operon responsible for the flagellar phase variation under different growth conditions in order to verify if its deletion is a frequent event that could explain the origin and dissemination of this serovar. In fact, coding sequences for transposons are present near this operon and in some strains, such as S. enterica Typhimurium LT2, the Fels-2 prophage gene is inserted near this operon. The presence of mobile DNA could confer instability to this region. In order to examine this, the cat (chloramphenicol acetyltransferase) gene was inserted adjacent to the fljBA operon so that deletions involving this genomic region could be identified. After growing S. enterica chloramphenicol-resistant strains under different conditions, more than 104 colonies were tested for the loss of chloramphenicol resistance. However, none of the colonies were sensitive to chloramphenicol. These data suggest that the origin of S. enterica serovar 4,5,12:i:- from Typhimurium by fljBA deletion is not a frequent event. The origin and dissemination of 4,5,12:i:- raise several questions about the role of flagellar phase variation in virulence.
McCabe, Evonne M; Burgess, Catherine M; O'Regan, Edel; McGuinness, Sheila; Barry, Thomas; Fanning, Séamus; Duffy, Geraldine
The objective of this study was the development of DNA and RNA real-time PCR methods for detection of food-borne Salmonella sp. as rapid alternatives to the traditional cultural method (ISO 6579, 2004) in fresh meat carcasses and processed meat samples. These PCR methods were based on the hilA sequence, with primers and hybridisation probes designed against this gene target. The primers and probes were evaluated for their efficiency and dynamic range and subsequently the specificity of the assay was tested using 106 Salmonella enterica subspecies enterica strains and 30 non-salmonellae strains. An internal amplification control (IAC) was also developed for incorporation. The optimum copy number of IAC was determined to be 500 copies per reaction. A complementary enrichment protocol was adapted from the existing standard ISO 6579:2004 and consisted of enrichment in Buffered Peptone Water (BPW) 22 ± 2 h and a second selective enrichment for 6 h in Rappaport Vassiliadis with Soya (RVS). The DNA and RNA-based real-time PCR protocols, were applied to meat samples inoculated with Salmonella enterica subspecies enterica strains, including swabs from meat carcasses and minced beef samples which were heat treated or frozen. The developed methods have the potential as useful alternatives to the standard ISO 6579:2004 method for the detection of Salmonella enterica subspecies enterica on carcass swabs and raw meat using hilA as a target. The DNA assay is a useful tool for the screening of meat samples in the abattoir within 3 days of slaughter or in a food production process and the RNA-based assay has the potential to detect viable Salmonella enterica subspecies enterica in ready-to-eat products. Copyright © 2010 Elsevier Ltd. All rights reserved.
Mukhopadhyay, Sudarsan; Sokorai, Kimberly; Ukuku, Dike; Fan, Xuetong; Juneja, Vijay; Sites, Joseph; Cassidy, Jennifer
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. Published by Elsevier B.V.
Hadjilouka, Agni; Andritsos, Nikolaos D; Paramithiotis, Spiros; Mataragas, Marios; Drosinos, Eleftherios H
The aim of this study was to assess serotype prevalence and biodiversity of Listeria monocytogenes strains isolated from diverse food products, i.e., minced pork, fruits, and vegetables. Three hundred twenty-six samples previously purchased from supermarkets and street markets within the Athens area were studied for L. monocytogenes prevalence. A total of 121 strains were isolated from the 36 samples that were positive for L. monocytogenes. Serotyping was performed with multiplex PCR, and biodiversity was assessed with random amplified polymorphic DNA (RAPD) PCR analysis using M13, UBC155, and HLWL85 as primers and with repetitive element palindromic (rep) PCR analysis using (GTG)5 as the primer. The majority (17 of 22) of the contaminated minced pork samples contained strains identified as serotype 1/2a, either alone or in combination with strains belonging to serotypes 1/2b, 4a, 4c, or 4ab. However, all L. monocytogenes isolates from fruits and vegetables belonged to serotype 4b. Rep-PCR provided better differentiation of the isolates than did RAPD PCR and resulted in discrimination of the isolates into a larger number of unique profiles. Complete differentiation was achieved only with the combination of these subtyping techniques.
Gay, Noellie; Le Hello, Simon; Weill, François-Xavier; de Thoisy, Benoit; Berger, Franck
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. Copyright © 2014 Elsevier B.V. All rights reserved.
van Asselt, E D; Thissen, J T N M; van der Fels-Klerx, H J
Salmonella serotype distribution can give insight in contamination routes and persistence along a production chain. Therefore, it is important to determine not only Salmonella prevalence but also to specify the serotypes involved at the different stages of the supply chain. For this purpose, data from a national monitoring program in the Netherlands were used to estimate the serotype distribution and to determine whether this distribution differs for the available sampling points in the broiler supply chain. Data covered the period from 2002 to 2005, all slaughterhouses (n = 22), and the following 6 sampling points: departure from hatchery, arrival at the farm, departure from the farm, arrival at the slaughterhouse, departure from the slaughterhouse, and end of processing. Furthermore, retail data for 2005 were used for comparison with slaughterhouse data. The following serotypes were followed throughout the chain: Salmonella Enteritidis, Salmonella Typhimurium, Salmonella Paratyphi B var. Java (Salmonella Java), Salmonella Infantis, Salmonella Virchow, and Salmonella Mbandaka. Results showed that serotype distribution varied significantly throughout the supply chain (P supply chain up to the retail phase.
Eller, Christoph; Simon, Sandra; Miller, Tatjana; Frick, Julia-Stefanie; Prager, Rita; Rabsch, Wolfgang; Guerra, Beatriz; Werner, Guido; Pfeifer, Yvonne
Between 20 000 and 35 000 cases of salmonellosis are detected annually in Germany, but only a few Salmonella are resistant to third-generation cephalosporins. The German National Reference Centre for Salmonella and other Enterics obtained 150 Salmonella enterica isolates from human infections between 2005 and 2011. In the present study we identified the β-lactamase genes causing resistance to third-generation cephalosporins in these isolates. For all isolates serotyping and antimicrobial susceptibility testing were performed. The presence of β-lactamase genes was detected by PCR amplification and sequencing. Isolates with identical serovar and β-lactamase genes were typed by XbaI macrorestriction followed by PFGE. Broth mate conjugation assays and plasmid analysis using S1 nuclease restriction of genomic DNA and subsequent PFGE as well as PCR-based replicon typing were performed for selected isolates. The 150 isolates were assigned to 30 different serovars, with S. enterica serovar Typhimurium (n = 73; 48.7%) as the most prevalent. Two different AmpC β-lactamase genes (blaCMY-2, n = 8; blaACC-1, n = 6) and various extended-spectrum β-lactamase (ESBL) genes were identified. The majority harboured the blaCTX-M-1 gene (n = 91; 60.7%) followed by blaCTX-M-14 (n = 12; 8.0%) and blaSHV-12 (n = 11; 7.3%). Typing of strains and subsequent comparison with selected Salmonella isolates from livestock revealed the presence of several clones in both humans and livestock. The wide spread of ESBL and AmpC genes in Salmonella of various serovars is most probably due to transfer of conjugative plasmids. Furthermore, our data indicate the clonal spread of distinct cephalosporin-resistant Salmonella strains from livestock to humans.
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
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. Copyright © 2016, American Society for Microbiology. All Rights Reserved.
Molina-López, Rafael A; Vidal, Anna; Obón, Elena; Martín, Marga; Darwich, Laila
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.
Prevalence of Salmonella enterica, Listeria monocytogenes, and pathogenic Escherichia coli in bulk tank milk and milk filters from US dairy operations in the National Animal Health Monitoring System Dairy 2014 study.
Sonnier, Jakeitha L; Karns, Jeffrey S; Lombard, Jason E; Kopral, Christine A; Haley, Bradd J; Kim, Seon-Woo; Van Kessel, Jo Ann S
The dairy farm environment is a well-documented reservoir for zoonotic pathogens such as Salmonella enterica, Shiga-toxigenic Escherichia coli, and Listeria monocytogenes, and humans may be exposed to these pathogens via consumption of unpasteurized milk and dairy products. As part of the National Animal Health Monitoring System Dairy 2014 study, bulk tank milk (BTM, n = 234) and milk filters (n = 254) were collected from a total of 234 dairy operations in 17 major dairy states and analyzed for the presence of these pathogens. The invA gene was detected in samples from 18.5% of operations and Salmonella enterica was isolated from 18.0% of operations. Salmonella Dublin was detected in 0.7% of operations. Sixteen Salmonella serotypes were isolated, and the most common serotypes were Cerro, Montevideo, and Newport. Representative Salmonella isolates (n = 137) were tested against a panel of 14 antimicrobials. Most (85%) were pansusceptible; the remaining were resistant to 1 to 9 antimicrobials, and within the resistant strains the most common profile was resistance to ampicillin/clavulanic acid, ampicillin, cefoxitin, ceftiofur, ceftriaxone, chloramphenicol, streptomycin, sulfisoxazole, and tetracycline. Listeria spp. were isolated from 19.9% of operations, and L. monocytogenes was isolated from 3.0% of operations. Serogroups 1/2a and 1/2b were the most common, followed by 4b and 4a. One or more E. coli virulence genes were detected in the BTM from 30.5% of operations and in the filters from 75.3% of operations. A combination of stx 2 , eaeA, and γ-tir genes was detected in the BTM from 0.5% of operations and in the filters from 6.6% of operations. The results of this study indicate an appreciable prevalence of bacterial pathogens in BTM and filters, including serovars known to infect humans. Copyright © 2018 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.
Baggesen, Dorte Lau; Wegener, Henrik Caspar
S. Typhimurium is one of the 2 most common salmonella serotypes causing human salmonellosis in Denmark. In order to illustrate the significance of different production animals as a source of infection, 1461 isolates were characterized by phage typing. The isolates originated from human patients a...
Ganguly, Arpeeta; Joerger, Rolf D
The open reading frames SEN0085 and SeKA_A4361, from Salmonella enterica serovar Enteritidis Nal R and serovar Kentucky 3795, respectively, corresponding to the acid-inducible sulfatase gene aslA from Salmonella enterica serovar Typhimurium, were previously suggested by microarray analysis to be differentially expressed under acid conditions. However, growth and enzyme activity tests in the present study demonstrated that both wild-type strains exhibited sulfatase activity with 4-nitrophenyl sulfate and 5-bromo-4-chloro-3 indolyl sulfate at pH 5.5. The acid sulfatase does not appear to be involved in sugar sulfate, tyrosine sulfate, 4-hydroxy-3-methoxyphenylglycol sulfate, heparin sulfate, or chondroitin sulfate hydrolysis at pH 5.5. Adhesion and invasion assays did not reveal differences between the serotypes and their corresponding aslA deletion mutants. Thus, the role and substrate(s) of AslA, a protein unique to salmonella and encoded in all sequenced Salmonella strains, remain elusive.
Takayama, Koichi; Reynolds, Paul N.; Short, Joshua J.; Kawakami, Yosuke; Adachi, Yasuo; Glasgow, Joel N.; Rots, Marianne G.; Krasnykh, Victor; Douglas, Joanne T.; Curiel, David T.
The efficiency of cancer gene therapy with recombinant adenoviruses based on serotype 5 (Ad5) has been limited partly because of variable, and often low, expression by human primary cancer cells of the primary cellular-receptor which recognizes the knob domain of the fiber protein, the coxsackie and adenovirus receptor (CAR). As a means of circumventing CAR deficiency, Ad vectors have been retargeted by utilizing chimeric fibers possessing knob domains of alternate Ad serotypes. We have reported that ovarian cancer cells possess a primary receptor for Ad3 to which the Ad3 knob binds independently of the CAR-Ad5 knob interaction. Furthermore, an Ad5-based chimeric vector, designated Ad5/3, containing a chimeric fiber proteins possessing the Ad3 knob, demonstrates CAR-independent tropism by virtue of targeting the Ad3 receptor. Based on these findings, we hypothesized that a mosaic virus possessing both the Ad5 knob and the Ad3 knob on the same virion could utilize either primary receptor, resulting in expanded tropism. In this study, we generated a dual-knob mosaic virus by coinfection of 293 cells with Ad5-based and Ad5/3-based vectors. Characterization of the resultant virions confirmed the incorporation of both Ad5 and Ad3 knobs in the same particle. Furthermore, this mosaic virus was able to utilize either receptor, CAR and the Ad3 receptor, for virus attachment to cells. Enhanced Ad infectivity with the mosaic virus was shown in a panel of cell lines, with receptor profiles ranging from CAR-dominant to Ad3 receptor-dominant. Thus, this mosaic virus strategy may offer the potential to improve Ad-based gene therapy approaches by infectivity enhancement and tropism expansion
Pollard, Dominic J; Young, Joanna C; Covarelli, Valentina; Herrera-León, Silvia; Connor, Thomas R; Fookes, Maria; Walker, Danielle; Echeita, Aurora; Thomson, Nicholas R; Berger, Cedric N; Frankel, Gad
Salmonella species utilize type III secretion systems (T3SSs) to translocate effectors into the cytosol of mammalian host cells, subverting cell signaling and facilitating the onset of gastroenteritis. In this study, we compared a draft genome assembly of Salmonella enterica subsp. salamae strain 3588/07 against the genomes of S. enterica subsp. enterica serovar Typhimurium strain LT2 and Salmonella bongori strain 12419. S. enterica subsp. salamae encodes the Salmonella pathogenicity island 1 (SPI-1), SPI-2, and the locus of enterocyte effacement (LEE) T3SSs. Though several key S Typhimurium effector genes are missing (e.g., avrA, sopB, and sseL), S. enterica subsp. salamae invades HeLa cells and contains homologues of S. bongori sboK and sboC, which we named seoC SboC and SeoC are homologues of EspJ from enteropathogenic and enterohemorrhagic Escherichia coli (EPEC and EHEC, respectively), which inhibit Src kinase-dependent phagocytosis by ADP-ribosylation. By screening 73 clinical and environmental Salmonella isolates, we identified EspJ homologues in S. bongori, S. enterica subsp. salamae, and Salmonella enterica subsp. arizonae The β-lactamase TEM-1 reporter system showed that SeoC is translocated by the SPI-1 T3SS. All the Salmonella SeoC/SboC homologues ADP-ribosylate Src E310 in vitro Ectopic expression of SeoC/SboC inhibited phagocytosis of IgG-opsonized beads into Cos-7 cells stably expressing green fluorescent protein (GFP)-FcγRIIa. Concurrently, S. enterica subsp. salamae infection of J774.A1 macrophages inhibited phagocytosis of beads, in a seoC-dependent manner. These results show that S. bongori, S. enterica subsp. salamae, and S. enterica subsp. arizonae share features of the infection strategy of extracellular pathogens EPEC and EHEC and shed light on the complexities of the T3SS effector repertoires of Enterobacteriaceae. Copyright © 2016 Pollard et al.
Schuchert, J.A.; Inzana, T.J.; Angen, Øystein
Multiplex PCR assays were developed to identify Actinobacillus pleuropneumoniae serotypes 1, 2, and 8. Primers designed for the conserved capsular polysaccharide (CP) export region amplified a 489-bp DNA fragment from all serotypes. Primers specific to the CP biosynthesis regions of serotypes 1, 2......, and 8 amplified fragments of 1.6 kb, 1.7 kb, and 970 bp from only their respective serotypes....
Kagambèga, Assèta; Lienemann, Taru; Frye, Jonathan G; Barro, Nicolas; Haukka, Kaisa
Multidrug-resistant Salmonella is an important cause of morbidity and mortality in developing countries. The aim of this study was to characterize and compare multidrug-resistant Salmonella enterica serovar Typhimurium isolates from patients and poultry feces. Salmonella strains were isolated from poultry and patients using standard bacteriological methods described in previous studies. The strains were serotype according to Kaufmann-White scheme and tested for antibiotic susceptibility to 12 different antimicrobial agents using the disk diffusion method. The whole genome of the S. Typhimurium isolates was analyzed using Illumina technology and compared with 20 isolates of S. Typhimurium for which the ST has been deposited in a global MLST database.The ResFinder Web server was used to find the antibiotic resistance genes from whole genome sequencing (WGS) data. For comparative genomics, publicly available complete and draft genomes of different S. Typhimurium laboratory-adapted strains were downloaded from GenBank. All the tested Salmonella serotype Typhimurium were multiresistant to five commonly used antibiotics (ampicillin, chloramphenicol, streptomycin, sulfonamide, and trimethoprim). The multilocus sequence type ST313 was detected from all the strains. Our sequences were very similar to S. Typhimurium ST313 strain D23580 isolated from a patient with invasive non-typhoid Salmonella (NTS) infection in Malawi, also located in sub-Saharan Africa. The use of ResFinder web server on the whole genome of the strains showed a resistance to aminoglycoside associated with carriage of the following resistances genes: strA , strB , and aadA1 ; resistance to β-lactams associated with carriage of a bla TEM-1B genes; resistance to phenicol associated with carriage of catA1 gene; resistance to sulfonamide associated with carriage of sul1 and sul2 genes; resistance to tetracycline associated with carriage of tet B gene; and resistance to trimethoprim associated to dfrA1 gene
Chávez-Navarro, Hilda; Hernández-Cueto, Daniel Dimitri; Vilchis-Estrada, Ariel; Bermúdez-Pulido, David César; Antonio-Andrés, Gabriela; Luria-Pérez, Rosendo
Salmonella enterica es una especie de bacterias anaeróbicas facultativas que han sido empleadas con gran éxito como vector bacteriano vivo atenuado con fines vacunales. Recientemente se ha documentado que S. enterica tiene propiedades importantes para ser considerada como agente terapéutico contra el cáncer. Estudios preclínicos y clínicos han demostrado que S. enterica coloniza tumores sólidos, semisólidos y metástasis, además de que contribuye a disminuir la resistencia a los tratamientos. ...
Nielsen, Eva Møller; Nielsen, Niels Ladefoged
from chicken products covered a large selection of serotypes. In contrast, the majority of the isolates from other product groups (turkey, poussin, wild birds) were concentrated on 1-3 serotypes. Using the standard procedure for antigen preparation and serotyping, 25 of the 156 strains (16%) were...
Thrane, Sandra Wingaard; Taylor, Véronique L.; Freschi, Luca
Introduction: Since the 1980’s the serotype O12 of Pseudomonas aeruginosa has emerged as the predominant serotype in clinical settings and in epidemic outbreaks. These serotype O12 isolates exhibit high levels of resistance to various classes of antibiotics.Methods: In this study, we explore how ......, and dangerous clones like O12 can be identified quickly....
Full Text Available Introduction. As one of mesophilic, easily cultivated species of pathogenic bacteria, Salmonella enterica transformed into viable but nonculturable (VNC state in response to environmental stresses, including action of biocides. The cells in this state, preserve the integrity of membranes and metabolism of some, but not detected by conventional methods of cultivation. Some researchers suggest that the evolutionary significance of this phenomenon is part of an adaptive response aimed at long-term survival of bacteria in adverse conditions; others argue that it is the result of stochastic cellular damage, in which nonculturable cells are in a state of gradual death. In any case, the phenomenon of existence VNC pathogens if they retain the ability to restore its growth in vivo is a significant problem in medicine, pharmaceutical, veterinary, food industry. VNC subpopulation of S. enterica was obtained under action of ethanol. In this paper was investigated in vivo resuscitation VNC S. enterica using intraperitoneal injection of mice. Materials and methods. Obtaining of stressful S. enterica populations. Bacteria were grown to exponential phase in broth Luria–Bertani (LB. To 1.0 ml sample suspension diluted to 1.5 × 106 cells/ml was added 1.0 ml of ethanol at a concentration of 40 % (v/v. After exposure of 10 to 600 minutes in the suspension were added 8.0 ml of phosphate buffered saline (FBS, washed by centrifugation (4500 g for 5 minutes and serially diluted at a ratio of 1:10 (v/v samples were stained with LIVE/DEAD BacLight (produced by "Invitrogen", USA, filtrated on membrane filters for fluorescence microscopy and parallel plated on LB agar cup to determine colony-forming units (CFU per ml. In vivo resuscitation VNC S. enterica was made following way. Three groups of animals were inoculated by intraperitoneal injection: 1 103 culturable cells (0.1 ml suspension containing 104 CFU / ml; 2 103 VNC cells (0.1 ml suspension containing 104 cells
P Scott Chandry
Full Text Available The genomic island encoding the locus of enterocyte effacement (LEE is an important virulence factor of the human pathogenic Escherichia coli. LEE typically encodes a type III secretion system (T3SS and secreted effectors capable of forming attaching and effacing lesions. Although prominent in the pathogenic E. coli such as serotype O157:H7, LEE has also been detected in Citrobacter rodentium, E. albertii, and although not confirmed, it is likely to also be in Shigella boydii. Previous phylogenetic analysis of LEE indicated the genomic island was evolving through stepwise acquisition of various components. This study describes a new LEE region from two strains of Salmonella enterica subspecies salamae serovar Sofia along with a phylogenetic analysis of LEE that provides new insights into the likely evolution of this genomic island. The Salmonella LEE contains 36 of the 41 genes typically observed in LEE within a genomic island of 49, 371 bp that encodes a total of 54 genes. A phylogenetic analysis was performed on the entire T3SS and four T3SS genes (escF, escJ, escN, and escV to elucidate the genealogy of LEE. Phylogenetic analysis inferred that the previously known LEE islands are members of a single lineage distinct from the new Salmonella LEE lineage. The previously known lineage of LEE diverged between islands found in Citrobacter and those in Escherichia and Shigella. Although recombination and horizontal gene transfer are important factors in the genealogy of most genomic islands, the phylogeny of the T3SS of LEE can be interpreted with a bifurcating tree. It seems likely that the LEE island entered the Enterobacteriaceae through horizontal gene transfer as a single unit, rather than as separate subsections, which was then subjected to the forces of both mutational change and recombination.
Sjölund-Karlsson, Maria; Howie, Rebecca L; Blickenstaff, Karen; Boerlin, Patrick; Ball, Takiyah; Chalmers, Gabhan; Duval, Brea; Haro, Jovita; Rickert, Regan; Zhao, Shaohua; Fedorka-Cray, Paula J; Whichard, Jean M
Non-Typhi Salmonella cause over 1.7 million cases of gastroenteritis in North America each year, and food-animal products are commonly implicated in human infections. For invasive infections, antimicrobial therapy is indicated. In North America, the antimicrobial susceptibility of Salmonella is monitored by the U.S. National Antimicrobial Resistance Monitoring System (NARMS) and The Canadian Integrated Program for Antimicrobial Resistance Surveillance (CIPARS). In this study, we determined the susceptibility to cephalosporins by broth microdilution among 5,041 non-Typhi Salmonella enterica isolated from food animals, retail meats, and humans. In the United States, 109 (4.6%) of isolates collected from humans, 77 (15.7%) from retail meat, and 140 (10.6%) from food animals displayed decreased susceptibility to cephalosporins (DSC). Among the Canadian retail meat and food animal isolates, 52 (13.0%) and 42 (9.4%) displayed DSC. All isolates displaying DSC were screened for β-lactamase genes (bla(TEM), bla(SHV), bla(CMY), bla(CTX-M), and bla(OXA-1)) by polymerase chain reaction. At least one β-lactamase gene was detected in 74/109 (67.9%) isolates collected from humans, and the bla(CMY) genes were most prevalent (69/109; 63.3%). Similarly, the bla(CMY) genes predominated among the β-lactamase-producing isolates collected from retail meats and food animals. Three isolates from humans harbored a bla(CTX-M-15) gene. No animal or retail meat isolates harbored a bla(CTX-M) or bla(OXA-1) gene. A bla(TEM) gene was found in 5 human, 9 retail meat, and 17 animal isolates. Although serotype distributions varied among human, retail meat, and animal sources, overlap in bla(CMY)-positive serotypes across sample sources supports meat and food-animal sources as reservoirs for human infection.
Dierickx, K; Pauwels, M; Laine, ML; Van Eldere, J; Cassiman, JJ; van Winkelhoff, AJ; van Steenberghe, D; Quirynen, M
Background: Porphyromonas gingivalis, a key pathogen in periodontitis, is able to adhere to and invade the pocket epithelium. Different capsular antigens of P gingivalis have been identified (K-serotyping). These P gingivalis capsular types show differences in adhesion capacity to human cell lines
Occurrence of foot and mouth disease serotypes in Tanzania: A retrospective study of tongue epithelial tissue samples. E. S. Swai, A. Mrosso, J. I.G. Masambu. Abstract. Samples of suspected foot and mouth disease (FMD) cases were collected via veterinary investigation centers (VIC) from different geographical locations ...
Sparding, Nadja; Dayie, Nicholas Tete Kwaku Dzifa; Mills, Richael O.
Streptococcus pneumoniae is a major cause of morbidity and mortality worldwide. Pneumococcal strains are classified according to their capsular polysaccharide and more than 90 different serotypes are currently known. In this project, three distinct groups of pneumococcal carriage isolates from Gh...
Barley yellow dwarf luteovirus (BYDV) serotypes PAV and RPV were identified from irrigated wheat (Triticum aestivum L.) samples from three provinces of Zambia by double antibody sandwich enzyme-linked immunosorbent assay using polyclonal and monoclonal antisera. Nine wheat cultivars were surveyed in 11 wheat ...
Objectives: The purpose of this study was to find out the sources and distribution of Salmonella serotypes isolated from apparently healthy slaughtered cattle and ... in food animals and meat products and underlines the necessity for a joint and coordinated surveillance and monitoring programs for salmonellosis and other ...
Conclusion: This study showed the diversity of Salmonella serotypes from both clinical and environmental samples and emergence of multiresistant Salmonella to antibiotics in Burkina Faso. A lettuce is a potential source of transmission of Salmonella causing diarrhea among human in Burkina Faso. Keywords: Salmonella ...
Dr. Cynthia Whitney, a CDC medical doctor and Epidemiologist, discusses serotype 12F pneumoniae. Created: 2/8/2018 by National Center for Emerging and Zoonotic Infectious Diseases (NCEZID). Date Released: 2/8/2018.
Objectives: To determine the serotype and antimicrobial susceptibility of E. coli in children under-five years of age. Methods: A cross-sectional study was conducted among 422 children with diarrhea from December 2011 to February 2012. Identification of E. coli and antimicrobial susceptibility testing were done following ...
Background: Diarrheal disease and its complications remain a major cause of morbidity and mortality in children. The prevalence and antibiogram of E. coli as causative agents of diarrhea vary from region to region, and even within countries in the same geographical area. Objectives: To determine the serotype and ...
Bello Gonzalez, Teresita; Rivera-Olivero, Ismar Alejandra; Sisco, María Carolina; Spadola, Enza; Hermans, Peter W; de Waard, Jacobus H
Serotype surveillance of Streptococcus pneumoniae is indispensable for evaluating the potential impact of pneumococcal conjugate vaccines. Serotyping by the standard Quellung reaction is technically demanding, time consuming, and expensive. A simple and economical strategy is multiplex PCR-based serotyping. We evaluated the cost effectiveness of a modified serial multiplex PCR (mPCR), resolving 24 serotypes in four PCR reactions and optimally targeting the most prevalent invasive and colonizing pneumococcal serotypes found in Venezuela. A total of 223 pneumococcal isolates, 140 invasive and 83 carriage isolates, previously serotyped by the Quellung reaction and representing the 18 most common serotypes/groups identified in Venezuela, were serotyped with the adapted mPCR. The mPCR serotyped 76% of all the strains in the first two PCR reactions and 91% after four reactions, correctly identifying 17 serotypes/groups. An isolate could be serotyped with mPCR in less than 2 minutes versus 15 minutes for the Quellung reaction, considerably lowering labor costs. A restrictive weakness of mPCR was found for the detection of 19F strains. Most Venezuelan 19F strains were not typeable using the mPCR, and two 19F cps serotype variants were identified. The mPCR assay is an accurate, rapid, and economical method for the identification of the vast majority of the serotypes from Venezuela and can be used in place of the standard Quellung reaction. An exception is the identification of serotype 19F. In this setting, most 19F strains were not detectable with mPCR, demonstrating a need of serology-based quality control for PCR-based serotyping.
Maurischat, Sven; Baumann, Beatrice; Martin, Annett; Malorny, Burkhard
Salmonella enterica is one of the most common zoonotic pathogens worldwide causing clinical diseases in human and animal hosts. Targeting a reduction of Salmonella prevalence in poultry, the EU set up a microbiological criterion that demands the absence of S. enterica subsp. enterica serovars Enteritidis and Typhimurium including its monophasic variant with seroformula 4,,12:i:- in 25 g of poultry neck skin samples and fresh meat according to regulation (EU) no 1086/2011. We developed and in-house validated a method that detects and differentiates these Salmonella serovars based on a 5-plex real-time PCR assay within 24 h after sampling. The inclusivity and exclusivity were between 98 and 99% analysing 456 bacterial strains. Validation according to ISO 16140:2003 against the traditional cultural reference method ISO 6579:2002 was performed using 60 artificially contaminated and 31 presumably naturally contaminated chicken neck skin samples resulting in a relative accuracy of 100%. The detection probability reached 100% between 3 and 5 CFU/25 g sample. We were also able to assign rough and non-motile strains to S. enterica subsp. enterica serovars Enteritidis and Typhimurium. In conclusion, we provide diagnostic laboratories a fast and accurate method to monitor these Salmonella serovars in chicken neck skin samples. Other matrices could be easily adapted. Copyright © 2014 Elsevier B.V. All rights reserved.
Full Text Available Over the last decades, several antimicrobial agents have been made available. Due to increasing antimicrobial resistance, bacteriophages were rediscovered for their potential applications against bacterial infections. In the present study, biofilm inhibition and eradication of Salmonella enterica subsp. enterica serovar Typhimurium DMC4 strain (S. Typhimurium was evaluated with respect to different incubation periods at different P22 phage titrations. The efficacy of P22 phage on biofilm formation and eradication of S. Typhimurium DMC4 strain was screened in vitro on polystyrene and stainless steel surfaces. The biofilm forming capacity of S. Typhimurium was significantly reduced at higher phage titrations (106 pfu/mL ≤. All phage titers (104-108 pfu/mL were found to be effective at the end of the 24 h-incubation period whereas higher phage titrations were found to be effective at the end of the 48 h and 72 h of incubation. P22 phage has less efficacy on already formed, especially mature biofilms (72 h-old biofilm. Notable results of P22 phage treatment on S. Typhimurium biofilm suggest that P22 phage has potential uses in food systems.
Batista, Diego Felipe Alves; de Freitas Neto, Oliveiro Caetano; de Almeida, Adriana Maria; Barrow, Paul Andrew; de Oliveira Barbosa, Fernanda; Berchieri Junior, Angelo
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. © 2016 The Author(s).
Perry, Malcolm B.; Angen, Øystein; MacLean, Leann L.
Atypical Actinobacillus pleuropneumoniae serotype 13 strains present in North America are described here for the first time. Different from serotype 13 strains described in Europe, North America strains are biotype I and antigenically related to both, serotypes 13 and 10. Chemical and structural...... and structurally identical with that of the reference strain of A. pleuropneumoniae serotype 10. The O-PS was characterized as a homopolymer of 1,2 linked β-d-galactofuranosyl residues, a structure unrelated to that of the O-PS produced by the reference strain of serotype 13. Strains from Canada and United States...
Valls Serón, M.; Haiko, J.; de Groot, P. G.; Korhonen, T. K.; Meijers, J. C. M.
Background: Pathogenic bacteria modulate the host coagulation system to evade immune responses or to facilitate dissemination through extravascular tissues. In particular, the important bacterial pathogens Salmonella enterica and Yersinia pestis intervene with the plasminogen/fibrinolytic system.
Full Text Available Salmonella enterica es una especie de bacterias anaeróbicas facultativas que han sido empleadas con gran éxito como vector bacteriano vivo atenuado con fines vacunales. Recientemente se ha documentado que S. enterica tiene propiedades importantes para ser considerada como agente terapéutico contra el cáncer. Estudios preclínicos y clínicos han demostrado que S. enterica coloniza tumores sólidos, semisólidos y metástasis, además de que contribuye a disminuir la resistencia a los tratamientos. En esta revisión se aborda la capacidad de S. enterica atenuada para eliminar células tumorales y su empleo como vector bacteriano vivo acarreador de moléculas heterólogas contra el cáncer.
Tapalski, D.; Hendriksen, Rene S.; Hasman, Henrik
This study describes the characterisation by pulsed-field gel electrophoresis (PFGE), multilocus variable number tandem repeat analysis (MLVA) typing and antimicrobial resistance profiles of 35 Salmonella enterica serovar Typhimurium isolates, mostly from infections in children who acquired...
Stege, H.; Christensen, Julia; Nielsen, J.P.
Our aim was to determine the Salmonella enterica prevalence in 96 randomly selected Danish pig herds, based on serological examination of blood samples and bacteriological examination of faecal samples (collected simultaneously from the same pens). For comparison, 39 high-seroprevalence herds were...... included in the study. The representativeness of the selected herds was assessed, based on descriptive statistics of herd size and type. Totals of 1330 pen samples and 6814 blood samples were examined. The results from the meat-juice screening in the Danish S. enterica Control Programme were available...... (average within-herd seroprevalence = 2%, maximum = 32%). (2) Among the 39 high-seroprevalence herds (recently assigned level 3 in the S. enterica Control Programme), S. enterica was isolated from 77% of the herds when 10 pen samples were examined bacteriologically. (3) Seropositivity tended to be related...
Li, Meng; Li, Mengzhe; Lin, Hong; Wang, Jingxue; Jin, Yanqiu; Han, Feng
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.
Full Text Available Abstract 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 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.
Christensen, Laurids Siig; Josefsen, Mathilde Hartmann; Pedersen, Karl
Free-range geese were sampled longitudinally and Salmonella isolates characterized to reveal highly diverging colonization dynamics. One flock was intermittently colonized with one strain of Salmonella enterica serovar Enteritidis from 2 weeks of age, while in another, S. enterica serovar Mbandaka...... appeared after 9 weeks, without dissemination but with multiple serovars appearing at later stages. ©American Society for Microbiology. All rights reserved....
Pickard, Derek; Wain, John; Baker, Stephen; Line, Alexandra; Chohan, Sonia; Fookes, Maria; Barron, Andrew; Gaora, Peadar O; Chabalgoity, José A; Thanky, Niren; Scholes, Christoph; Thomson, Nicholas; Quail, Michael; Parkhill, Julian; Dougan, Gordon
Vi capsular polysaccharide production is encoded by the viaB locus, which has a limited distribution in Salmonella enterica serovars. In S. enterica serovar Typhi, viaB is encoded on a 134-kb pathogenicity island known as SPI-7 that is located between partially duplicated tRNA(pheU) sites. Functional and bioinformatic analysis suggests that SPI-7 has a mosaic structure and may have evolved as a consequence of several independent insertion events. Analysis of viaB-associated DNA in Vi-positive S. enterica serovar Paratyphi C and S. enterica serovar Dublin isolates revealed the presence of similar SPI-7 islands. In S. enterica serovars Paratyphi C and Dublin, the SopE bacteriophage and a 15-kb fragment adjacent to the intact tRNA(pheU) site were absent. In S. enterica serovar Paratyphi C only, a region encoding a type IV pilus involved in the adherence of S. enterica serovar Typhi to host cells was missing. The remainder of the SPI-7 islands investigated exhibited over 99% DNA sequence identity in the three serovars. Of 30 other Salmonella serovars examined, 24 contained no insertions at the equivalent tRNA(pheU) site, 2 had a 3.7-kb insertion, and 4 showed sequence variation at the tRNA(pheU)-phoN junction, which was not analyzed further. Sequence analysis of the SPI-7 region from S. enterica serovar Typhi strain CT18 revealed significant synteny with clusters of genes from a variety of saprophytic bacteria and phytobacteria, including Pseudomonas aeruginosa and Xanthomonas axonopodis pv. citri. This analysis suggested that SPI-7 may be a mobile element, such as a conjugative transposon or an integrated plasmid remnant.
Full Text Available Severe evolution of pneumococcal infections with multiresistant strains in children under 2 years of age determined the introduction, in some countries, of the heptavalent vaccine, which includes the most frequent capsular serotypes. The knowledge of serotypes circulating in our area is crucial for the introduction of such a vaccine in our country. We studied 202 pneumococcal strains; out of these, serologic identification of 172 strains established classification in 23 serotypes/15 serogroups; 24 strains were non-typable. 66,3% of isolates belong to serotypes 23F/23B, 6B/6A and 19F/19A. Only 54% of the serotypes isolated from children under 2 years of age are included in the heptavalent vaccine. Pneumococcal strains with high level resistance to beta-lactams and multiresistant to other antibiotics belong to the 2 most frequently isolated serotypes, 19A and 23B. Vaccinal serotypes 4 and 18C were not identified in our study.
Erol, Irfan; Goncuoglu, Muammer; Ayaz, Naim Deniz; Ellerbroek, Lüppo; Ormanci, Fatma Seda Bilir; Kangal, Ozlem Iseri
The aim of the study was to find out the serotype distribution of 169 Salmonella colonies recovered from 112 Salmonella positive ground turkey (115 colonies) and 52 turkey meat parts (54 colonies). Out of 15 Salmonella serotypes: S. Corvallis, S. Kentucky, S. Bredeney, S. Virchow, S. Saintpaul and S. Agona were identified as the predominant serovars at the rates of 27%, 13%, 12%, 12%, 11%, and 10%, respectively. Other serotypes were below 6% of the total isolates. All S. Kentucky and S. Virchow and most of the S. Corvallis (39/46) and S. Heidelberg (9/9) serotypes were recovered from ground turkey. The results indicate that turkey ground meat and meat parts were contaminated with quite distinct Salmonella serotypes. This is the first study reporting Salmonella serotype distribution in turkey meat and S. Corvallis as predominant serotype in poultry meat in Turkey.
Full Text Available The aim of the study was to find out the serotype distribution of 169 Salmonella colonies recovered from 112 Salmonella positive ground turkey (115 colonies and 52 turkey meat parts (54 colonies. Out of 15 Salmonella serotypes: S. Corvallis, S. Kentucky, S. Bredeney, S. Virchow, S. Saintpaul and S. Agona were identified as the predominant serovars at the rates of 27%, 13%, 12%, 12%, 11%, and 10%, respectively. Other serotypes were below 6% of the total isolates. All S. Kentucky and S. Virchow and most of the S. Corvallis (39/46 and S. Heidelberg (9/9 serotypes were recovered from ground turkey. The results indicate that turkey ground meat and meat parts were contaminated with quite distinct Salmonella serotypes. This is the first study reporting Salmonella serotype distribution in turkey meat and S. Corvallis as predominant serotype in poultry meat in Turkey.
Evangelopoulou, Grammato; Kritas, Spyridon; Govaris, Alexander; Burriel, Angeliki R
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.
Full Text Available Rapid detection of food-borne pathogens is essential to public health and the food industry. Although the conventional culture method is highly sensitive, it takes at least a few days to detect food-borne pathogens. Even though polymerase chain reaction (PCR can detect food-borne pathogens in a few hours, it is more expensive and unsatisfactorily sensitive relative to the culture method. We have developed a method to rapidly detect Salmonella enterica by using a compact disc (CD-shaped device that can reduce reagent consumption in conventional PCR. The detection method, which combines culture and PCR, is more rapid than the conventional culture method and is more sensitive and cheaper than PCR. In this study, we also examined a sample preparation method that involved collecting bacterial cells from food. The bacteria collected from chicken meat spiked with S. enterica were mixed with PCR reagents, and PCR was performed on the device. At a low concentration of S. enterica, the collected S. enterica was cultured before PCR for sensitive detection. After cultivation for 4 h, S. enterica at 1.7 × 104 colony-forming units (CFUs·g−1 was detected within 8 h, which included the time needed for sample preparation and detection. Furthermore, the detection of 30 CFUs·g−1 of S. enterica was possible within 12 h including 8 h for cultivation.
Buchovec, I; Vaitonis, Z; Luksiene, Z
Salmonellosis is one of the most common foodborne diseases in the world. The aim of this study was to evaluate the antibacterial efficiency of 5-aminolevulinic acid (ALA) based photosensitization against one of food pathogens Salmonella enterica. Salmonella enterica was incubated with ALA (7.5 mmol l(-1)) for 1-4 h and afterwards illuminated with visible light. The light source used for illumination of S. enterica emitted light lambda = 400 nm with energy density 20 mW cm(-2). The illumination time varied from 0 to 20 min and subsequently a total energy dose reached 0-24 J cm(-2). The data obtained indicate that S. enterica is able to produce endogenous photosensitizer PpIX when incubated with ALA. Remarkable inactivation of micro-organisms can be achieved (6 log) after photosensitization. It is obvious that photosensitization-based inactivation of S. enterica depends on illumination as well as incubation with ALA time. ALA-based photosensitization can be an effective tool against multi-drug resistant Gram-negative bacteria S. enterica serovar Typhimurium. Experimental data and mathematical evaluations support the idea that ALA-based photosensitization can be a useful tool for the development of nonthermal food preservation technology in future.
Le Thomas, I; Lepercq, J; Bergeret, M; Francoual, C; Raymond, J
The classification of serogroup B streptococci in serotype is based on the structural differences of capsular polysaccharides and on presence or absence of a protein c antigen. They are classified as Ia, Ia/c, Ib/c, II, II/c, III, IV and V. The serotype V, unknown in 1970, seems emerging, and is placed in third position of frequency in some American studies. We have therefore decided to evaluate its frequency in Paris. In a population of 137 pregnant women and 60 neonates carrying streptococcus of serogroup B, the serotype was systematically determined using the test "Group B streptococcus serotyping test" (Dako, Danemark). In the pregnant women population, 12% of the isolated strains were of serotype V, 26% of serotype III, 15% of serotype II, 14% of serotype Ia, and 21% could not be typed. In neonates, it represented 15% of the isolates and took place after the serotype Ia (20%), the serotype III (18%) and the serotype II (15%). None of the neonates had early- or late-onset disease. They were only colonized. Only one mother exhibited, during the per-partum, a positive blood culture with a streptococcus group B of serotype V. These results confirm, in Paris, the importance of this serotype previously observed in foreign studies. It represents 11 to 15% of the isolated streptococcus group B in the neonates and can cause early or late-onset disease. However, larger studies are needed to evaluate the exact risk of pathology for the serotype V and its significance in neonatal infectious disease.
Cooper, Caitlin; Moore, Robert J; Chandry, P Scott; Fegan, Narelle
1. Salmonella enterica subsp. salamae serovar sofia (S. sofia) is a prevalent strain of Salmonella in Australian broilers and has been isolated from broiler chickens, litter, dust, as well as pre- and post-processing carcasses, and retail chicken portions but has never been reported in commercial Australian layers or eggs. 2. To investigate whether a S. sofia isolate from a broiler could colonise layers, one-month-old Hyline brown layers were orally inoculated with S. sofia and colonisation was monitored for 2-4 weeks. 3. Overall, 30-40% of the chickens shed S. sofia from the cloaca between 6 and 14 d post-inoculation which then declined to 10% by d 21. Necropsy at 2 weeks post-inoculation revealed 80% of birds harboured S. sofia in the caecum, whilst, by 4 weeks post-infection, no chickens were colonised with S. sofia in the gastrointestinal tract, liver or spleen. Additionally, no aerosol 'bird to bird' transfer was evident. 4. This study demonstrated that laying hens can be colonised by broiler-derived S. sofia; however, this colonisation was transient, reaching a peak at 14 d post-inoculation, and was completely cleared by 28 d post-inoculation. The transience of colonisation of S. sofia in layers could be a factor explaining why S. sofia has never been detected when screening for Salmonella serotypes found in Australian laying hens or eggs.
Jean-Gilles Beaubrun, Junia; Flamer, Marie-Laure; Addy, Nicole; Ewing, Laura; Gopinath, Gopal; Jarvis, Karen; Grim, Chris; Hanes, Darcy E
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. Published by Elsevier Ltd.
Russo, Elizabeth T; Biggerstaff, Gwen; Hoekstra, R Michael; Meyer, Stephanie; Patel, Nehal; Miller, Benjamin; Quick, Rob
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
Jelsbak, Lotte; Thomsen, Line Elnif; Wallrodt, Inke
Sensing and responding to environmental cues is a fundamental characteristic of bacterial physiology and virulence. Here we identify polyamines as novel environmental signals essential for virulence of Salmonella enterica serovar Typhimurium, a major intracellular pathogen and a model organism......, we show that an S. Typhimurium polyamine mutant is defective for invasion, intracellular survival, killing of the nematode Caenorhabditis elegans and systemic infection of the mouse model of typhoid fever. Virulence of the mutant could be restored by genetic complementation, and invasion...... and intracellular survival could, as well, be complemented by the addition of exogenous putrescine and spermidine to the bacterial cultures prior to infection. Interestingly, intracellular survival of the polyamine mutant was significantly enhanced above the wild type level by the addition of exogenous putrescine...
Jijón, Steffani; Wetzel, Amy; LeJeune, Jeffrey
Between May and September 2004, fecal samples from various wildlife species admitted to two rehabilitation centers in Ohio were cultured for Salmonella enterica and Escherichia coli O157:H7. Eight of 71 (11%) samples, including specimens from three opossums (Didelphis virginiana), two gray squirrels (Sciurus carolinensis), a woodchuck (Marmota monax), a Harris hawk (Parabuteo unicinctus), and a screech owl (Otus asio) tested positive for Salmonella serovars Braenderup, Senftenberg, Oranienburg, and Kentucky. The Salmonella Oranienburg isolates were indistinguishable by pulsed-field gel electrophoresis. Most isolates were susceptible to commonly used antibiotics; however, the Salmonella Kentucky isolate was resistant to multiple beta-lactam antibiotics (amoxicillin/clavulanic acid and ampicillin), cefoxitin, and ceftiofur, a third-generation cephalosporin. Escherichia coli O157:H7 was not isolated from any sample. Transmission of Salmonella from wildlife may occur between animals at rehabilitation centers.
Thong, Kwai Lin; Ngoi, Soo Tein; Chai, Lay Ching; Teh, Cindy Shuan Ju
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.
Mkrtchyan, M.S.; Zakaryan, М. K.; Mnatsakanyan, А. А.
.Enteritidis). Previously, we reported that the induction of the cytokine network and an antimicrobial protein is serotype-specific and also depends on the disease stage. Differential genomic context of the serotypes may explain the differential induction of inflammatory responses. Recent studies have indicated...... that bacterial infections in early life may help to inhibit excessive allergic Th2 reactions by angling the immune system towards Th1 responses. However, it is known that infections can also cause the exacerbation of allergic reactions. Skewing of response away from Treg cells may lead to the onset and....../or progression of autoimmune diseases in humans. It is also thought that there is a high probability that infectious gastroenteritis increases the risk of subsequent autoimmune and allergic diseases....
Ashshi, Ahmed Mohamed
Transmission of dengue virus (DENV) through blood transfusion has been documented and hence screening for DENV during blood donation has been recently recommended by the American Association of Blood Banks and Centres of Disease Control and Prevention. DENV is endemic in the Western province of the Kingdom of Saudi Arabia (KSA) and serotypes 1, 2 and 3, but not 4, have been detected. However, little is known regarding the rates of DENV during blood donation in the kingdom. The aim of this study was therefore to measure the prevalence of dengue virus and its serotypes in eligible Saudi blood donors in the endemic Western region of KSA. This was a cross-sectional study and serum samples were collected from 910 eligible Saudi male blood donors. DENV IgM and IgG antibodies were measured serologically by ELISA while viral serotypes were detected by a single step IVD CE certified multiplex RT-PCR kit. The overall prevalence was 39 and 5.5% for IgG+ and IgM+, respectively. There were 12 (1.3%) with exclusively IgM+, 317 (34.8%) exclusively IgG+ and 38 (4.2%) with dual IgM+/IgG+ donors. The overall prevalence was 3.2% (n = 29) and 2.3% (n = 21) for primary and secondary infections. PCR was positive in 5.5% (n = 50) and, DENV-2 (n = 24; 48%) was the most frequent serotype and was significantly higher than DENV-1 (20%; P = 0.02) and DENV-3 (2%; P = 0.1 × 10 -5 ) but not DENV-4 (30%; P = 0.2). There was no significant difference between both DENV-4 and DENV-1 (P = 0.4). The combination of the PCR and serology findings showed that 22 (2.4%) and 28 (3.1%) donors had primary and secondary viremic infections, respectively. The detected rates of DENV by PCR suggest a potential high risk of viral transmission by blood transfusion. To the best of our knowledge, this study is the first to report the detection of DENV-4 serotype in Saudi Arabia. More studies are required to measure the precise prevalence of DENV serotypes and their potential
Thrane, Sandra Wingaard; Taylor, Véronique L.; Freschi, Luca
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 cli...
Feenstra, F.; Maris-Veldhuis, M.A.; Daus, F.J.; Tacken, M.G.J.; Moormann, R.J.M.; Gennip, van H.G.P.; Rijn, van P.A.
Bluetongue virus (BTV) causes Bluetongue in ruminants and is transmitted by Culicoides biting midges. Vaccination is the most effective measure to control vector borne diseases; however, there are 26 known BTV serotypes showing little cross protection. The BTV serotype is mainly determined by genome
Spanjaard, L.; Bol, P.; de Marie, S.; Zanen, H. C.
Case histories of 692 patients with meningococcal disease due to serogroup B, C, or W (W-135) were reviewed to study the association of the serotypes 2a and 2b with the course of disease. The case-fatality rate in group B disease was significantly associated with serotype 2b (B:2b) strains (P =
Samir K Saha
Full Text Available BACKGROUND: Streptococcus pneumoniae is a leading cause of meningitis in countries where pneumococcal conjugate vaccines (PCV targeting commonly occurring serotypes are not routinely used. However, effectiveness of PCV would be jeopardized by emergence of invasive pneumococcal diseases (IPD caused by serotypes which are not included in PCV. Systematic hospital based surveillance in Bangladesh was established and progressively improved to determine the pathogens causing childhood sepsis and meningitis. This also provided the foundation for determining the spectrum of serotypes causing IPD. This article reports an unprecedented upsurge of serotype 2, an uncommon pneumococcal serotype, without any known intervention. METHODS AND FINDINGS: Cases with suspected IPD had blood or cerebrospinal fluid (CSF collected from the beginning of 2001 till 2009. Pneumococcal serotypes were determined by capsular swelling of isolates or PCR of culture-negative CSF specimens. Multicenter national surveillance, expanded from 2004, identified 45,437 patients with suspected bacteremia who were blood cultured and 10,618 suspected meningitis cases who had a lumber puncture. Pneumococcus accounted for 230 culture positive cases of meningitis in children <5 years. Serotype-2 was the leading cause of pneumococcal meningitis, accounting for 20.4% (45/221; 95% CI 15%-26% of cases. Ninety eight percent (45/46 of these serotype-2 strains were isolated from meningitis cases, yielding the highest serotype-specific odds ratio for meningitis (29.6; 95% CI 3.4-256.3. The serotype-2 strains had three closely related pulsed field gel electrophoresis types. CONCLUSIONS: S. pneumoniae serotype-2 was found to possess an unusually high potential for causing meningitis and was the leading serotype-specific cause of childhood meningitis in Bangladesh over the past decade. Persisting disease occurrence or progressive spread would represent a major potential infection threat since serotype-2
Huhtamo, E; Korhonen, Em; Vapalahti, O
Imported dengue cases originating from the Madeiran outbreak are increasingly reported. In 2012 five Finnish travellers returning from Madeira were diagnosed with dengue fever. Viral sequence data was obtained from two patients. The partial C-preM sequences (399 and 396 bp respectively) were found similar to that of an autochthonous case from Madeira. The partial E-gene sequence (933 bp) which was identical among the two patients grouped phylogenetically with South American strains of dengue virus serotype 1.
M.H. Chew; M.M. Rahman; J. Jelip; M.R. Hassan; I. Isahak
Dengue is a severe disease caused by dengue virus (DENV), transmitted to human being by infected Aedes mosquitoes. It is a major public health concern in Southeast Asia due to its fatality in the form of hemorrhagic fever (DHF) and dengue shock syndrome (DSS). The objective of the study was to isolate and identify dengue virus serotypes prevalent in endemic areas of Kuala Lumpur and Selangor in Malaysia by virus culture, indirect immunoflurecent assay and molecular techniques. A total number ...
Lopes, Graciela Volz; Pissetti, Caroline; da Cruz Payão Pellegrini, Débora; da Silva, Luis Eduardo; Cardoso, Marisa
Salmonella enterica subsp. enterica plays a role as a foodborne pathogen worldwide. The consumption of contaminated pork has been associated with human salmonellosis and the increase in antimicrobial resistance among Salmonella from pigs and pork products is a concern. A total of 225 Salmonella isolates from feed mills, the lairage environment, and the intestinal contents of pigs and carcasses were investigated for their antimicrobial susceptibility. A MIC for ciprofloxacin was screened by agar dilution, and antimicrobial resistance genes were investigated by PCR assays. Among the tested isolates, 171 (76%) showed resistance to at least one antimicrobial agent, and 91 (40.4%) were multiresistant. Resistance occurred most frequently to tetracycline (54.5%), sulfonamides (39.6%), and streptomycin (33.7%). Thirty-two (94.1%) nalidixic acid-resistant isolates exhibited decreased susceptibility to ciprofloxacin. The resistance genes found were blaTEM (ampicillin), tet(A) (tetracycline), tet(B) (tetracycline/minocycline), sul1, sul2, and sul3 (sulfonamides), catA1 (chloramphenicol), floR (florfenicol/chloramphenicol), strA and strB (streptomycin), aph(3')-Ia (kanamycin), aac(3)-IIa and aac(3)-IVa (apramycin/gentamicin), aadA variant (streptomycin/spectinomycin), and dfrA1 (trimethoprim). Salmonella isolates from pig feces and carcasses displayed a higher frequency of resistance to most antimicrobials tested than isolates from feed mills. Common resistance gene profiles were found in isolates from the lairage and the intestinal content of pigs and carcasses, demonstrating that resistance genes selected on farms may be found in pork.
Hurd, H S; Gailey, J K; McKean, J D; Griffith, R W
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 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-hold group compared to the other two groups. The mean Minolta L and the drip loss were significantly higher in the short-hold group. From this and other studies, it appears that elimination of the holding process is not feasible S. enterica control option, given current U.S. harvesting systems.
Narender S Maan
Full Text Available Bluetongue (BT is an arthropod-borne viral disease, which primarily affects ruminants in tropical and temperate regions of the world. Twenty six bluetongue virus (BTV serotypes have been recognised worldwide, including nine from Europe and fifteen in the United States. Identification of BTV serotype is important for vaccination programmes and for BTV epidemiology studies. Traditional typing methods (virus isolation and serum or virus neutralisation tests (SNT or VNT are slow (taking weeks, depend on availability of reference virus-strains or antisera and can be inconclusive. Nucleotide sequence analyses and phylogenetic comparisons of genome segment 2 (Seg-2 encoding BTV outer-capsid protein VP2 (the primary determinant of virus serotype were completed for reference strains of BTV-1 to 26, as well as multiple additional isolates from different geographic and temporal origins. The resulting Seg-2 database has been used to develop rapid (within 24 h and reliable RT-PCR-based typing assays for each BTV type. Multiple primer-pairs (at least three designed for each serotype were widely tested, providing an initial identification of serotype by amplification of a cDNA product of the expected size. Serotype was confirmed by sequencing of the cDNA amplicons and phylogenetic comparisons to previously characterised reference strains. The results from RT-PCR and sequencing were in perfect agreement with VNT for reference strains of all 26 BTV serotypes, as well as the field isolates tested. The serotype-specific primers showed no cross-amplification with reference strains of the remaining 25 serotypes, or multiple other isolates of the more closely related heterologous BTV types. The primers and RT-PCR assays developed in this study provide a rapid, sensitive and reliable method for the identification and differentiation of the twenty-six BTV serotypes, and will be updated periodically to maintain their relevance to current BTV distribution and
Salmonella enterica subsp. enterica bacteria are important foodborne pathogens with major economic impact. Some isolates exhibit increased heat tolerance, a concern for food safety. Analysis of a finished-quality genome sequence of an isolate commonly used in heat resistance studies, S. enterica sub...
Use of multiple-locus variable-number of tandem repeats analysis (MLVA) to investigate genetic diversity of Salmonella enterica subsp. enterica serovar Typhimurium isolates from human, food, and veterinary sources
Mateva, Gergana; Pedersen, Karl; Sørensen, Gitte
Salmonella enterica subspecies enterica serovar Typhimurium is the most common zoonotic pathogen in Bulgaria. To allow efficient outbreak investigations and surveillance in the food chain, accurate and discriminatory methods for typing are needed. This study evaluated the use of multiple-locus va...
Muthuirulandi Sethuvel, Dhiviya Prabaa; Devanga Ragupathi, Naveen Kumar; Anandan, Shalini; Walia, Kamini; Veeraraghavan, Balaji
It is not always possible to identify Shigella serogroups/serotypes by biochemical properties alone. Specific identification requires serotyping. Occasionally, isolates that resemble Shigella spp. biochemically, but are non-agglutinable with available antisera, have been observed. Several mechanisms have been reported to limit the efficiency of the serotyping assay. Serotype conversion is a major mechanism in Shigella spp. to escape protective host immune responses. This easy conversion through significant modification of the O-antigen backbone results in different serotypes, which makes laboratory identification difficult. Furthermore, members of the family Enterobacteriaceae are closely related and there is antigenic cross-over (intra- and inter-specific cross-reaction) which affects the agglutination reaction. The performance of the available methods for identification of non-serotypeable Shigella is discussed here, and reveals them to be non-reliable. This shows a need for an alternative method for identification and typing of Shigella spp.
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
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
Full Text Available 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.
Kiely, R A
This study determined the carriage rate and serotype distribution of group B Streptococcus (GBS) in women of child-bearing age in the southern region of Ireland. A total of 2000 vaginal swabs collected in two periods in 2004 and 2006 were examined and revealed a GBS carriage rate of 16·1%. Serotyping of isolates showed that serotypes Ia, II, III, IV, and V were the most prevalent. A high prevalence of serotype IV was found, increasing from 7·6% to 15·2% between 2004 and 2006. Random amplified polymorphic DNA analysis demonstrated considerable genetic heterogeneity in the serotype IV isolates. This serotype should be considered for inclusion in potential vaccines for use in Ireland.
The number of food poisoning cases caused by enteropathogens has increased in recent years. A significant part of the outbreaks associated with the consumption of raw vegetables has been attributed to Escherichia coli O157:H7 and Salmonella enterica subsp. enterica serovar Typhimurium. Bovine manure
Franz, E.; Bruggen, van A.H.C.
There is an increased concern that plants might be more important as a carrier for human enteric pathogens like E. coli O157:H7 and Salmonella enterica serovars than previously thought. This review summarizes the knowledge available on the ecology of E. coli O157:H7 and Salmonella enterica in the
Iriarte, Andrés; Giner-Lamia, Joaquín; Silva, Claudia; Betancor, Laura; Astocondor, Lizeth; Cestero, Juan J; Ochoa, Theresa; García, Coralith; Puente, José L; Chabalgoity, José A; García-Del Portillo, Francisco
We report a 4.99-Mb draft genome sequence of Salmonella enterica subsp. enterica serovar Infantis strain SPE101, isolated from feces of a 5-month-old breast-fed female showing diarrhea associated with severe dehydration and malnutrition. The infection prolonged for 6 months despite antibiotic treatment. Copyright © 2017 Iriarte et al.
Fischer, Nicholas J
This article describes a notable case of Haemophilus influenzae serotype a (Hia) septic arthritis in an immunized central Australian indigenous child. Since the widespread immunization for H. influenzae serotype b (Hib) in many indigenous peoples worldwide, there has been an increase in reported cases of Hia, postulating that this serotype is taking over the niche that Hib once occupied in indigenous populations. Copyright © 2014 The Authors. Published by Elsevier Ltd.. All rights reserved.
inhibitors of Clostridium botulinum neurotoxin serotype A (BoNT/A). Virtual screening was initially performed by computationally docking com- pounds of the...species Clostridium botulinum , C. baratii, and C. butyricum, consist of seven immunologically distinct serotypes (A to G). BoNTs are synthesized as 150...4. TITLE AND SUBTITLE Identification and biochemical characterization of small-molecule inhibitors of Clostridium botulinum neurotoxin serotype
Full Text Available Introduction: Porcine pleuropneumonia inflicts important economic losses on most commercial herds. Detection of subclinical or chronic infection in animals still remains a challenge, as isolation and identification of A. pleuropneumoniae serotypes is difficult and quantification of the bacteria on agar plates is often almost impossible. The aim of the study was to develop and evaluate a serotype-specific quantitative TaqMan probe-based PCR for detection of serotype 2 in pig lungs, tonsils, and nasal swabs.
Kendall, Brian A; Dascomb, Kristin K; Mehta, Rajesh R; Stockmann, Chris; Mason, Edward O; Ampofo, Krow; Pavia, Andrew T; Byington, Carrie L
Pneumococcal conjugate vaccines (PCV) have indirect effects due to decreased Streptococcus pneumoniae colonization in vaccine recipients. We sought to determine whether the introduction of PCV13 in children led to changes in the epidemiology and clinical manifestations of invasive pneumococcal disease (IPD) in adults. We described demographics, comorbidities, clinical manifestations, and serotypes of IPD in Utah adults before (November 2009-February 2010) and after (March 2010-March 2012) the introduction of PCV13 in children. We also compare serotypes causing IPD in Utah adults and children. After the introduction of PCV13 in the childhood vaccine program, the proportion of IPD due to PCV13 exclusive serotypes decreased significantly in Utah adults (64-40%, p=0.009), primarily due to a decline in serotype 7F (36-15%, p=0.008). There were non-significant increases in IPD due to Pneumococcal polysaccharide 23 (PPV23) unique serotypes and non-vaccine serotypes, most notably serotype 22F. Changes in the proportions of vaccine and non-vaccine serotypes were similar in adults and children. Meningitis was more commonly due to non-vaccine serotypes relative to non-meningitis cases (47% vs. 18%, p=0.007). When stratified by sex, decreases in PCV13 serotype IPD were only noted in men (76-33%, p=0.001). Serotype epidemiology of IPD in adults closely follows that of children in the PCV13 era. Continued surveillance is needed to confirm whether replacement serotypes will lead to increases in pneumococcal meningitis and whether there are sex differences in the indirect effects of PCV vaccination in children. Copyright © 2015 Elsevier Ltd. All rights reserved.
Irfan Erol; Muammer Goncuoglu; Naim Deniz Ayaz; Lüppo Ellerbroek; Fatma Seda Bilir Ormanci; Ozlem Iseri Kangal
The aim of the study was to find out the serotype distribution of 169 Salmonella colonies recovered from 112 Salmonella positive ground turkey (115 colonies) and 52 turkey meat parts (54 colonies). Out of 15 Salmonella serotypes: S. Corvallis, S. Kentucky, S. Bredeney, S. Virchow, S. Saintpaul and S. Agona were identified as the predominant serovars at the rates of 27%, 13%, 12%, 12%, 11%, and 10%, respectively. Other serotypes were below 6% of the total isolates. All S. Kentucky and S. V...
Janaina Viana da Rosa
Full Text Available Microorganisms that cause human diseases can contaminate fishes in aquatic environments as well as during their capture, handling, and transport. The purpose of this study was to isolate Vibrio parahaemolyticus and Salmonella enterica from fishes captured in the Lagoa dos Patos estuary that were eviscerated and cleaned for trade. Thirteen fish landing were analyzed and 65 entire fishes and 65 cleaned fishes were studied to determine the presence of V. parahaemolyticus and S. enterica. Bacterial isolates were compared using rep-PCR. V. parahaemolyticus was isolated from one entire Micropogonias furnieri and two entire Mugil platanus, as well as from three eviscerated M. platanus. S. enterica was isolated from two eviscerated Paralichthys orbignyanus. Identical rep-PCR bands from V. parahaemolyticus were observed in entire and eviscerated fishes from the same discharge, suggesting processing failures that neither eliminated the microorganism from the raw material nor prevented cross-contamination. S. enterica was not isolated from entire fishes, presumably because contamination occurred due to hygiene and sanitary failures. Our results showed that M. furnieri and M. platanus captured in the Lagoa dos Patos estuary may host V. parahaemolyticus and that this microorganism, as with S. enterica, may also persist even after the fish is cleaned. This is the first record of the isolation of V. parahaemolyticus from M. furnieri.
Lienau, E. Kurt; Blazar, Jeffrey M.; Wang, Charles; Brown, Eric W.; Stones, Robert; Musser, Steven; Allard, Marc W.
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
Bugarel, M; Tudor, A; Loneragan, G H; Nightingale, K K
Foodborne illnesses due to Salmonella represent an important public-health concern worldwide. In the United States, a majority of Salmonella infections are associated with a small number of serotypes. Furthermore, some serotypes that are overrepresented among human disease are also associated with multi-drug resistance phenotypes. Rapid detection of serotypes of public-health concern might help reduce the burden of salmonellosis cases and limit exposure to multi-drug resistant Salmonella. We developed a two-step real-time PCR-based rapid method for the identification and detection of five Salmonella serotypes that are either overrepresented in human disease or frequently associated with multi-drug resistance, including serotypes Enteritidis, Typhimurium, Newport, Hadar, and Heidelberg. Two sets of four markers were developed to detect and differentiate the five serotypes. The first set of markers was developed as a screening step to detect the five serotypes; whereas, the second set was used to further distinguish serotypes Heidelberg, Newport and Hadar. The utilization of these markers on a two-step investigation strategy provides a diagnostic specificity of 97% for the detection of Typhimurium, Enteritidis, Heidelberg, Infantis, Newport and Hadar. The diagnostic sensitivity of the detection makers is >96%. The availability of this two-step rapid method will facilitate specific detection of Salmonella serotypes that contribute to a significant proportion of human disease and carry antimicrobial resistance. Published by Elsevier B.V.
Bailey, J S; Fedorka-Cray, P J; Stern, N J; Craven, S E; Cox, N A; Cosby, D E
The subtyping and identification of bacterial pathogens throughout food processing and production chains is useful to the new hazard analysis critical control point-based food safety plans. Traditional manual serotyping remains the primary means of subtyping Salmonella isolates. Molecular biology techniques, however, offer the promise of more rapid and sensitive subtyping of Salmonella. This study evaluates the potential of restriction enzyme PvuII, followed by probing with the rRNA operon from Escherichia coli, to generate serotype-specific DNA fingerprints. A total of 32 identified serotypes were found with an overall agreement in 208 of the 259 (80%) isolates tested between U.S. Department of Agriculture serotype identification and riboprint serotype identification. Many of the isolates that did not correlate were serotype identified as Salmonella Montevideo, which indicates that for this serotype, there are multiple ribotypes. When Salmonella Montevideo isolates were not included, the ribotype identification agreed with serotyping in 207 of the 231 (90%) isolates. The primary outcome of any ribotyping procedure is to give distinct ribotype patterns. This extensive poultry epidemiological study demonstrates that, in addition to ribotype patterns, the identification of isolates to known serotypes provides the investigator with additional information that can be more useful than traditional epidemiology and isolate identification studies.
Gatti, B M; Ramirez Gronda, G A; Etchevarría, M; Vescina, C M; Varea, A M; González Ayala, S E
Haemophilus influenzae (Hi) is the causative agent of several human diseases such as sepsis, meningitis, celulitis, and osteoarthritis. We investigated the isolation of Hi serotypes from sterile sites in sick children. One hundred and seventy nine strains from 146 patients were studied, period 1996-2002, at the Microbiology Laboratory, Hospital de Niños Superiora Sor María Ludovica, Argentina. The serotype distribution was:1 a, 112 b,1 c,1 d, 4 e, 3 f y 24 no typable. Since the beginning of universal Hi b vaccination in 1998, we have observed the fast decrease of serotype b and a relative increase of other serotypes.
Shif, I; Silberstein, I; Aboudy, Y; Mendelson, E; Mates, A; Gotlieb-Stematsky, T
Each of three consecutive cold seasons (November-March) in the town of Tiberias, Israel, was dominated by one particular rotavirus serotype causing acute diarrhoea in the community: the 1987/88 season by serotype-2; 1988/89 by serotype-1 and 1989/90 by serotype-4. Each season was also characterized by a particular pattern of rotaviral RNA when visualized using electrophoresis in gels. RNA profiles of identical rota serotypes and serotypic prevalence for any given cold season were unique for the town of Tiberias and different from other localities throughout Israel. The meaning of these findings in terms of herd immunity is discussed.
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...
Anup Kollanoor Johny
Full Text Available Background:Salmonella Enteritidis phage type 8 (PT8 is a major poultry-associated Salmonella strain implicated in foodborne outbreaks in the United States. We previously reported that two plant-derived compounds generally recognized as safe (GRAS, trans-cinnamaldehyde (TC, and eugenol (EG, significantly reduced S. Enteritidis colonization in broiler and layer chickens. To elucidate potential PT8 genes affected by TC and EG during colonization, a whole-genome microarray analysis of the bacterium treated with TC and EG was conducted.Results:S. Enteritidis PT8 was grown in Luria-Bertani broth at 37°C to an OD600 of ~0.5. Subinhibitory concentrations (SICs; concentration that does not inhibit bacterial growth of TC (0.01%; 0.75 mM or EG (0.04%; 2.46 mM were then added to the culture. S. Enteritidis PT8 RNA was extracted before and 30 min after TC or EG addition. Labeled cDNA from three replicate experiments was subsequently hybridized to a microarray of over 99% of S. Enteritidis PT4 genes, and the hybridization signals were quantified. The plant-derived compounds down-regulated (P < 0.005 expression of S. Enteritidis PT8 genes involved in flagellar motility, regulation of the Salmonella Pathogenicity Island 1, and invasion of intestinal epithelial cells. TC and EG also suppressed transcription of genes encoding multiple transport systems and outer membrane proteins. Moreover, several metabolic and biosynthetic pathways in the pathogen were down-regulated during exposure to the plant-derived compounds. Both TC and EG stimulated the transcription of heat shock genes, such as dnaK, dnaJ, ibpB, and ibpA in S. Enteritidis PT8 (P < 0.005. The results obtained from microarray were validated using a quantitative real-time PCR.Conclusion: The plant-derived compounds TC and EG exert antimicrobial effects on S. Enteritidis PT8 by affecting multiple genes, including those associated with virulence, colonization, cell membrane composition, and transport systems.
Laconcha, I.; Baggesen, Dorte Lau; Rementeria, A.
profile A10-A10-A1 was predominant and specific for this phage type. It is concluded that PFGE, in combination with phage typing, represents a suitable tool for the epidemiological typing of Salmonella Enteritidis strains which could be used for investigations or surveillance of the international spread......A total of 101 strains of Salmonella Enteritidis phage types (PT) 1, 4, 6, and 8 from Denmark, England and Spain were studied by PFGE to elucidate genetic relationships among strains isolated from animal, human and environmental sources between 1983 and 1997. Analysis with Xba I, Bin I and Spe I......, confirming the close relationship among these phage types and the protracted spread of a single clone over a large geographical area. In general, strains from Denmark showed more variation in their PFGE profiles than those from England and Spain. PT4 strains exhibited genetic homogeneity in the three...
Kermas, Rachida; Touati, Abdelaziz; Brasme, Lucien; Le Magrex-Debar, Elisabeth; Mehrane, Sadjia; Weill, François-Xavier; De Champs, Christophe
The purpose of this work was to study the genetic determinants responsible for extended-spectrum cephalosporin (ESC) resistance of Salmonella collected during the period of 1995-2008 at the Hussein Dey hospital in Algiers (Algeria). Fourteen ESC-resistant Salmonella isolates were tested towards 22 antimicrobial agents. Polymerase chain reaction (PCR) and sequencing were used to determine the underlying genetic determinants responsible for the extended-spectrum beta-lactamase (ESBL) phenotypes. Enterobacterial Repetitive Intergenic Consensus PCR was employed to type the isolates. All tested isolates were resistant to ticarcillin, ticarcillin-clavulanate, piperacillin, cefuroxime, aztreonam, ceftazidime, cefotaxime (except two isolates), cefepime, and cefpirome. PCR and DNA sequencing identified these ESBLs as TEM-48 (n=6), TEM-4 (n=3), CTX-M-15 (n=4), and one new TEM, designated TEM-188. Thus, continued surveillance for the presence of ESBL-producing (non-typhoidal) salmonellae in Algeria is essential.
Background: Salmonella Enteritidis phage type 8 (PT8) is a major poultry-associated Salmonella strain implicated in foodborne outbreaks in the United States. We previously reported that two GRAS-status, plant-derived compounds, trans-cinnamaldehyde (TC) and eugenol (EG) significantly reduced S. Ent...
Full Text Available The live vaccine Cevac S. Gallinarum, made from a rough strain of Salmonella enterica subspecies enterica serotype Gallinarum is used for preventing fowl typhoid, a disease that still causes considerable economic losses in countries with a developing poultry industry. The objective of this paper was to evaluate a possible reversion to virulence of the strain used in a vaccine in commercial brown layers. Only Salmonella-free chicks were utilized. One hundred twenty (120 12-day-old Dekalb brown layers divided in two trials were used. The first trial had six groups of 15 birds each. Birds of group 1 were vaccinated with 10 doses of Cevac S. Gallinarum subcutaneously and 10 doses orally, in a total of 20 doses of vaccine. Then the birds of groups 2, 3, 4, and 5 received inocula that contained feces and a pool of organs with fragments of liver, heart, spleen, and cecal tonsils obtained from the immediately previous group. The second trial had three groups with 10 birds each. Birds in group 7 received inocula containing a pool of organs from birds of group 5 from trial 1, whilst the birds in group 8 were vaccinated subcutaneously with one dose of vaccine. Both trials included negative control groups (6 and 9. Throughout the experimental period, birds were monitored for reactions to the vaccination on the site of administration, clinical signs, and post-mortem lesions. In each passage, in addition to the birds euthanized to provide the inocula material, two birds from each group were euthanized for assessment of possible lesions, and their organs (liver, heart, spleen and cecal tonsils were cultured in an attempt to isolate the vaccine strain. Except for one bird from group 1, that had a local reaction on the site of vaccination - a small vesicle with less that 0.5 mm that persisted until the third day post vaccination -, no other bird had any local reaction to the vaccine or any visible clinical alteration. Birds in group 8 did not present any
Mousing, Jan; Jensen, P.T.; Halgaard, C.
A nation-wide Salmonella enterica surveillance and control programme was initiated in Danish finishing herds over the first quarter of 1995, In Denmark, all swine for slaughter are identifiable by a unique herd code. For each herd code, and depending on the herd's annual kill, random samples...... herd sizes). Irrespective of Salmonella level, all herds receive a monthly update on the current results of the S. enterica test results. IF a herd is categorized in Level 2 or 3, it must receive an advisory visit by a practising veterinarian and a local swine extension specialist, and certain...... management hygiene precautions must be taken. If a herd is categorized in Level 3, the finishers from the herd must additionally be slaughtered under special hygiene precautions. This is supervised by the veterinary authorities. During 1995, 604 000 samples were tested for S. enterica, corresponding to 3...
Full Text Available 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.
Ana Victoria Garcia
Full Text Available Infections with Salmonella enterica belong to the most prominent causes of food poisoning and infected fruits and vegetables represent important vectors for salmonellosis. Whereas 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, the data gathered suggest that Salmonella triggers PTI in plants and evolved strategies to avoid or subvert plant immunity.
Korhonen, T K
Pla of the plague bacterium Yersinia pestis and PgtE of the enteropathogen Salmonella enterica are surface-exposed, transmembrane β-barrel proteases of the omptin family that exhibit a complex array of interactions with the hemostatic systems in vitro, and both proteases are established virulence factors. Pla favors fibrinolysis by direct activation of plasminogen, inactivation of the serpins plasminogen activator inhibitor-1 and α2-antiplasmin, inactivation of the thrombin-activable fibrinolysis inhibitor, and activation of single-chain urokinase. PgtE is structurally very similar but exhibits partially different functions and differ in expression control. PgtE proteolysis targets control aspects of fibrinolysis, and mimicry of matrix metalloproteinases enhances cell migration that should favor the intracellular spread of the bacterium. Enzymatic activity of both proteases is strongly influenced by the environment-induced variations in lipopolysaccharide that binds to the β-barrel. Both proteases cleave the tissue factor pathway inhibitor and thus also express procoagulant activity. © 2015 International Society on Thrombosis and Haemostasis.
García, Ana V.
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. 2014 Garca and Hirt.
de Toro, María; Sáenz, Yolanda; Cercenado, Emilia; Rojo-Bezares, Beatriz; García-Campello, Marta; Undabeitia, Esther; Torres, Carmen
The mechanisms of antimicrobial resistance were characterized in 90 Salmonella enterica isolates either resistant or with intermediate resistance to amoxicillin/clavulanate (AMC(R/I)) or resistant to third-generation cephalosporins (C3G(R)). These isolates were recovered in three Spanish hospitals during 2007-2009. The C3G(R) phenotype was expressed by three isolates that carried the following extended-spectrum β-lactamase genes: phage-associated bla(CTX M-10) in S. Virchow, bla(CTX-M-14a) surrounded by ISEcp1 and IS903 in S. Enteritidis, and bla(CTX-M-15) linked to ISEcp1 and orf477 in S. Gnesta (first description in this serotype). The AMC(R/I) phenotype was found in 87 isolates (79 S. Typhimurim, 7 S. Enteritidis, and one S. Thompson). The bla(PSE-1) gene, followed by bla(OXA-1) was mostly found among S. Typhimurim, and the bla(TEM-1) gene among S. Enteritidis. Three different gene combinations [bla(PSE-1) +floR+aadA2+sul+tet(G); bla(OXA-1) +catA+aadA1/strA-strB+sul+tet(B) and bla(TEM-1) + cmlA1+aadA/strA-strB+sul+tet(A)/tet(B) genes] were associated with the ampicillin-chloramphenicol-streptomycin-sulfonamides-tetracycline phenotype in 68 AMC(R/I) S. enterica isolates. Class 1 integrons were observed in 79% of the isolates and in most of them (45 isolates) two integrons including the aadA2 and bla(PSE-1) gene cassettes, respectively, were detected. The bla(OXA-1) +aadA1 arrangement was detected in 23 isolates, and the aac(6')-Ib-cr+bla(OXA-1) +catB3+arr3 in another one. Non-classic class 1 integrons were found in three isolates: dfrA12+orfF+aadA2+cmlA1+aadA1 (1 isolate), dfrA12+orfF+aadA2+ cmlA1+aadA1+qacH+IS440+sul3 (1 isolate) and dfrA12+orfF+aadA2+cmlA1+aadA1+qacH+IS440+ sul3+orf1+mef(B)Δ-IS26 (1 isolate). Taken together, these results underline the need for clinical concern regarding β-lactam resistance in Salmonella and thus for continuous monitoring.
González Morales, N L; Núñez-López, M; Ramos-Castañeda, J; Velasco-Hernández, J X
In this work we present a mathematical model that incorporates two Dengue serotypes. The model has been constructed to study both the epidemiological trends of the disease and conditions that allow coexistence in competing strains under vaccination. We consider two viral strains and temporary cross-immunity with one vector mosquito population. Results suggest that vaccination scenarios will not only reduce disease incidence but will also modify the transmission dynamics. Indeed, vaccination and cross immunity period are seen to decrease the frequency and magnitude of outbreaks but in a differentiated manner with specific effects depending upon the interaction vaccine and strain type. Copyright © 2016 Elsevier Inc. All rights reserved.
Newton, Richard; Wernisch, Lorenz
Streptococcus pneumoniae is a human pathogen that is a major cause of infant mortality. Identifying the pneumococcal serotype is an important step in monitoring the impact of vaccines used to protect against disease. Genomic microarrays provide an effective method for molecular serotyping. Previously we developed an empirical Bayesian model for the classification of serotypes from a molecular serotyping array. With only few samples available, a model driven approach was the only option. In the meanwhile, several thousand samples have been made available to us, providing an opportunity to investigate serotype classification by machine learning methods, which could complement the Bayesian model. We compare the performance of the original Bayesian model with two machine learning algorithms: Gradient Boosting Machines and Random Forests. We present our results as an example of a generic strategy whereby a preliminary probabilistic model is complemented or replaced by a machine learning classifier once enough data are available. Despite the availability of thousands of serotyping arrays, a problem encountered when applying machine learning methods is the lack of training data containing mixtures of serotypes; due to the large number of possible combinations. Most of the available training data comprises samples with only a single serotype. To overcome the lack of training data we implemented an iterative analysis, creating artificial training data of serotype mixtures by combining raw data from single serotype arrays. With the enhanced training set the machine learning algorithms out perform the original Bayesian model. However, for serotypes currently lacking sufficient training data the best performing implementation was a combination of the results of the Bayesian Model and the Gradient Boosting Machine. As well as being an effective method for classifying biological data, machine learning can also be used as an efficient method for revealing subtle biological
Li, Baoguang; Vellidis, George; Liu, Huanli; Jay-Russell, Michele; Zhao, Shaohua; Hu, Zonglin; Wright, Anita; Elkins, Christopher A
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. Copyright © 2014, American Society for
Pickard, Derek; Wain, John; Baker, Stephen; Line, Alexandra; Chohan, Sonia; Fookes, Maria; Barron, Andrew; Ó Gaora, Peadar; Chabalgoity, José A.; Thanky, Niren; Scholes, Christoph; Thomson, Nicholas; Quail, Michael; Parkhill, Julian; Dougan, Gordon
Vi capsular polysaccharide production is encoded by the viaB locus, which has a limited distribution in Salmonella enterica serovars. In S. enterica serovar Typhi, viaB is encoded on a 134-kb pathogenicity island known as SPI-7 that is located between partially duplicated tRNApheU sites. Functional and bioinformatic analysis suggests that SPI-7 has a mosaic structure and may have evolved as a consequence of several independent insertion events. Analysis of viaB-associated DNA in Vi-positive S...
Colavecchio, Anna; D’Souza, Yasmin; Tompkins, Elizabeth; Jeukens, Julie; Freschi, Luca; Emond-Rheault, Jean-Guillaume; Kukavica-Ibrulj, Irena; Boyle, Brian; Bekal, Sadjia; Tamber, Sandeep; Levesque, Roger C.; Goodridge, Lawrence D.
Salmonella enterica is a bacterial species that is a major cause of illness in humans and food-producing animals. S. enterica exhibits considerable inter-serovar diversity, as evidenced by the large number of host adapted serovars that have been identified. The development of methods to assess genome diversity in S. enterica will help to further define the limits of diversity in this foodborne pathogen. Thus, we evaluated a PCR assay, which targets prophage integrase genes, as a rapid method to investigate S. enterica genome diversity. To evaluate the PCR prophage integrase assay, 49 isolates of S. enterica were selected, including 19 clinical isolates from clonal serovars (Enteritidis and Heidelberg) that commonly cause human illness, and 30 isolates from food-associated Salmonella serovars that rarely cause human illness. The number of integrase genes identified by the PCR assay was compared to the number of integrase genes within intact prophages identified by whole genome sequencing and phage finding program PHASTER. The PCR assay identified a total of 147 prophage integrase genes within the 49 S. enterica genomes (79 integrase genes in the food-associated Salmonella isolates, 50 integrase genes in S. Enteritidis, and 18 integrase genes in S. Heidelberg). In comparison, whole genome sequencing and PHASTER identified a total of 75 prophage integrase genes within 102 intact prophages in the 49 S. enterica genomes (44 integrase genes in the food-associated Salmonella isolates, 21 integrase genes in S. Enteritidis, and 9 integrase genes in S. Heidelberg). Collectively, both the PCR assay and PHASTER identified the presence of a large diversity of prophage integrase genes in the food-associated isolates compared to the clinical isolates, thus indicating a high degree of diversity in the food-associated isolates, and confirming the clonal nature of S. Enteritidis and S. Heidelberg. Moreover, PHASTER revealed a diversity of 29 different types of prophages and 23
Full Text Available Tetracyclines are broad-spectrum agents, exhibiting activity against a wide range of gram-positive and gram-negative bacteria and are currently used for therapy and prophylaxis of human infections and for the prevention and control of bacterial infections in veterinary medicine. Streptomycin has only limited current usage in clinical medicine, but this antibiotic remains important for therapeutic and growth promotion in animals and for the bacterial disease control in plants.The increasing incidence of resistance to streptomycin and tetracyclines has been reported worldwide in Salmonella spp. of human and animal origin. Fifty-eight multidrug-resistant Salmonella enterica strains of twenty different serotypes, were chosen among the collection of multidrug-resistant strains isolated from animals and food of animal origin at the Istituto Zooprofilattico Sperimentale delle Venezie and at the Istituto Zooprofilattico Sperimentale dell’Abruzzo e Molise, during their routine surveillance activity in the 2000 and 2001 period.All strains showed resistance to at least three different antimicrobials: tetracycline and streptomycin resistances represent the 98% and 95% of the strains, respectively. Salmonella isolates were analyzed for the presence of genetic determinants conferring streptomycin and tetracyclines resistance by PCR for the tet(A and strA-strB genes, respectively.The strA-strB genes were highly prevalent in Salmonella strains of our collection, being detected in the 83% of the streptomycin resistant strains; the 68% of the tetracycline resistant strains were tet(A gene positive, indicating that this gene is widely diffused in Salmonella strains circulating in animals in Italy. Two prevalent repN- and repI1-resistance plasmids were identified in Salmonella isolates of our collection. In many strains, the strA-strB genes were linked to a particular Tn5393-derivative transposon, characterized by the presence of the insertion sequence IS1133
Baggesen, Dorte Lau; Wegener, Henrik Caspar
S. Typhimurium is one of the 2 most common salmonella serotypes causing human salmonellosis in Denmark. In order to illustrate the significance of different production animals as a source of infection, 1461 isolates were characterized by phage typing. The isolates originated from human patients...... and from cattle, pigs and poultry. By phage typing the isolates could be separated in 35 different phage types. Five types (10, 12, 66, 110 and 135) predominated and comprised 78.8% of the isolates. In humans, 57.3% of the isolates were phage type 12. This phage type was also predominant in pig herds and......, to a lesser degree, in cattle. Phage types 110, 120, 135 and 193 constituted 86.5% of the poultry isolates while these phage types only made up 12.9% of the human isolates. The investigation showed that pigs are probably a major source of S. Typhimurium infection in humans in Denmark today....
Park, Saeyoung; Gerber, Sabina; Lee, Jean C.
Most Staphylococcus aureus isolates produce either a serotype 5 (CP5) or 8 (CP8) capsular polysaccharide, and the CP antigens are targets for vaccine development. Since CP5 and CP8 have similar trisaccharide repeating units, it is important to identify an epitope shared by both CP5 and CP8. To characterize cross-reactivity between CP5 and CP8, the immunogenicity of CP5 and CP8 conjugate vaccines in mice and rabbits was evaluated by serological assays. Immune sera were also tested for function...
Nielsen, R.; Andresen, Lars Ole; Plambeck, Tamara
). Immunodiffusion confirmed the antigenic relationship with serotype 2 and further demonstrated an antigenic relationship with strain WF83 (reference strain of serotype 7). SDS-PAGE with LPS from strains 1536, 4226, WF83 and strain 7317 (representative of the 9 isolates examined) showed that strains WF83 and 7317...
Drolet, B.S.; Reister, L.M.; Mecham, J.O.; Wilson, W.C.; Nol, P.; Vercauteren, K.C.; Rijn, van P.A.; Bowen, R.A.
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
Hatrongjit, Rujirat; Kerdsin, Anusak; Gottschalk, Marcelo; Takeuchi, Dan; Hamada, Shigeyuki; Oishi, Kazunori; Akeda, Yukihiro
Streptococcus suis is a zoonotic pathogen that causes invasive infections in humans and pigs. It has been reported that S. suis infection in humans is mostly caused by serotype 2. However, human cases caused by other serotypes have rarely been reported. This is the first report of a human case of infection with S. suis serotype 31 in Thailand. A 55-year-old male alcohol misuser with liver cirrhosis was admitted with sepsis to a hospital in the Central Region of Thailand. He had consumed a homemade, raw pork product prior to the onset of illness. He was alive after treatment with ceftriaxone and no complication occurred. An isolate from blood culture at the hospital was suspected as viridans group Streptococcus. It was confirmed at a reference laboratory as S. suis serotype 31 by biochemical tests, 16S rDNA sequencing, and multiplex polymerase chain reaction for serotyping, but it was untypable by the co-agglutination test with antisera against recognized S. suis serotypes, suggesting loss of capsular material. The absence of a capsule was confirmed by transmission electron microscopy. The isolate was confirmed to be sequence type 221, with 13 putative virulence genes that are usually found in serotype 2 strains. We should be aware of the emergence of S. suis infections caused by uncommon serotypes in patients with predisposing conditions. Laboratory capacity to identify S. suis in the hospital is needed in developing countries, which can contribute to enhanced surveillance, epidemiological control, and prevention strategies in the prevalent area.
Long, M; Lai, H; Deng, W; Zhou, K; Li, B; Liu, S; Fan, L; Wang, H; Zou, L
The study aimed to serotype the Salmonella isolates recovered from chicken and egg production chains, and to investigate the disinfectant resistance phenotypes and genotypes of these isolates. The Salmonella isolates were serotyped, and the minimal inhibitory concentrations (MICs) of disinfectants were determined. Results showed that the Salmonella isolates recovered from both chains were diverse, and the serotypes in each part of the production chain and between the two production chains were significantly different. In the chicken production chain, 19 different serotypes were recovered, while only five serotypes were found in the egg production chain. The isolates showed a high susceptibility to didecyldimethylammonium bromide (DDAB) but a low susceptibility to benzalkonium chloride (BC), benzalkonium bromide (BAB) and chlorhexidine (CHX). Salmonella Enteritidis and Salmonella Typhimurium were more resistant to BC and BAB. The qacEΔ1 and qacF resistance genes were detected in 26·7 and 7·7% of the isolates respectively. The qacEΔ1 gene was frequently found in Salmonella Derby and Salm. Enteritidis (P < 0·05). Our findings indicated that Salmonella was commonly present in both chains, and could serve as a critical vector in spreading disinfectant resistance associated with different serotypes. This study first demonstrated disinfectant resistance phenotypes and genotypes of serotyped Salmonella. The study highlights the need for monitoring the disinfectant resistance varied in different Salmonella serotypes. © 2016 The Society for Applied Microbiology.
Marinho, Paula Eillanny Silva; Bretas de Oliveira, Danilo; Candiani, Talitah Michel Sanchez; Crispim, Ana Paula Correia; Alvarenga, Pedro Paulo Martins; Castro, Fabrizia Cristina Dos Santos; Abrahão, Jonatas Santos; Rios, Maria; Coimbra, Roney Santos; Kroon, Erna Geessien
To determine the causes of viral meningitis, we analyzed 22 cerebrospinal fluid samples collected during the 2014-2015 dengue epidemics in Brazil. We identified 3 serotypes of dengue virus (DENV-1, -2, and -3), as well as co-infection with 2 or 3 serotypes. We also detected the Asian II genotype of DENV-2.
Bruun, B; Gahrn-Hansen, B; Westh, H
Surveillance performed after the introduction of general Haemophilus influenzae serotype b (Hib) vaccination in Denmark identified 13 cases of invasive bacteraemic H. influenzae serotype f (Hif) disease in adults over a period of 7 years. Bacteraemic respiratory tract infections accounted for 61...
Garzetti, Debora; Susen, Rosa; Fruth, Angelika; Tietze, Erhard; Heesemann, Jürgen; Rakin, Alexander
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. Copyright © 2013 Elsevier GmbH. All rights reserved.
Qi, Wei; Zeng, Yuhong; Orgel, Scott; Francon, Alain; Kim, Jae Hyun; Randolph, Theodore W; Carpenter, John F; Middaugh, C Russell
To improve the effectiveness of the polio vaccination campaign, improvements in the thermal stability of the vaccine are being investigated. Here, inactivated polio vaccine, serotype 3 (IPV3) was characterized via a number of biophysical techniques. The size was characterized by transmission electronic microscopy and light scattering. The capsid protein conformation was evaluated by intrinsic fluorescence and circular dichroism (CD), and the D-antigen content by enzyme-linked immunosorbent assay (ELISA). The pH thermal stability of IPV3 (pH 3.0-8.0; 10°C-87.5°C) was evaluated by fluorescence, CD, and static light scattering. The transition temperatures reflect the responses, respectively, of tertiary structure, secondary structure, and size to applied thermal stress. The data were summarized as empirical phase diagrams, and the most stable conditions were found to be pH 7.0 with temperature lower than 40°C. CD detected a higher transition temperature for capsid protein than that for RNA. The effects of certain excipients on IPV3 thermal stability and antigen content were evaluated. The results of their effects, based on intrinsic fluorescence and ELISA, were in good agreement, suggesting the feasibility of applying intrinsic fluorescence as a high-throughput tool for formulation development. The study improves the understanding of IPV3 thermal stability, and provides a starting point for future formulation development of IPV3 and other serotypes. © 2013 Wiley Periodicals, Inc. and the American Pharmacists Association.
Full Text Available Recombinant, replication-deficient serotype 5 adenovirus infects the liver upon in vivo, systemic injection in rodents. This infection requires the binding of factor X to the capsid of this adenovirus. Another organ, the adrenal gland is also infected upon systemic administration of Ad, however, whether this infection is dependent on the cocksackie adenovirus receptor (CAR or depends on the binding of factor X to the viral capsid remained to be determined. In the present work, we have used a pharmacological agent (warfarin as well as recombinant adenoviruses lacking the binding site of Factor X to elucidate this mechanism in mice. We demonstrate that, as observed in the liver, adenovirus infection of the adrenal glands in vivo requires Factor X. Considering that the level of transduction of the adrenal glands is well-below that of the liver and that capsid-modified adenoviruses are unlikely to selectively infect the adrenal glands, we have used single-photon emission computed tomography (SPECT imaging of gene expression to determine whether local virus administration (direct injection in the kidney could increase gene transfer to the adrenal glands. We demonstrate that direct injection of the virus in the kidney increases gene transfer in the adrenal gland but liver transduction remains important. These observations strongly suggest that serotype 5 adenovirus uses a similar mechanism to infect liver and adrenal gland and that selective transgene expression in the latter is more likely to be achieved through transcriptional targeting.
Full Text Available The multidimensional nature of dengue virus (DENV infections, which can be caused by four distinct serotypes of the virus, complicates the sensitivity of assays designed for the diagnosis of infection. Different viral markers can be optimally detected at different stages of infection. Of particular clinical importance is the early identification of infection, which is pivotal for disease management and the development of blood screening assays. Non-structural protein 1 (NS1 is an early surrogate marker of infection and its detection in serum coincides with detectable viraemia. The aim of this work was to isolate and characterise serotype-specific monoclonal antibodies that bind to NS1 for each of the four DENV serotypes. This was achieved using phage display and a subtractive biopanning strategy to direct the antibody selection towards serotype-specific epitopes. This antibody isolation strategy has advantages over immunisation techniques where it is difficult to avoid antibody responses to cross-reactive, immunodominant epitopes. Serotype specificity to recombinant antigen for each of the antibodies was confirmed by Enzyme Linked Immunosorbent Assay (ELISA and Surface Plasmon Resonance. Confirmation of binding to native DENV NS1 was achieved using ELISA and immunofluorescence assay on DENV infected Vero cells. No cross-reactivity with Zika or Kunjin viruses was observed. A previously isolated pan-reactive antibody that binds to an immunodominant epitope was able to pair with each of the serotype-specific antibodies in a sandwich ELISA, indicating that the serotype specific antibodies bind to epitopes which are all spatially distinct from the immunodominant epitope. These antibodies were suitable for use in a multiplexed assay for simultaneous detection and serotyping of DENV NS1 in human serum. This work demonstrates that phage display coupled with novel biopanning strategies is a valuable in vitro methodology for isolation of binders that can
Lebani, Kebaneilwe; Jones, Martina L; Watterson, Daniel; Ranzoni, Andrea; Traves, Renee J; Young, Paul R; Mahler, Stephen M
The multidimensional nature of dengue virus (DENV) infections, which can be caused by four distinct serotypes of the virus, complicates the sensitivity of assays designed for the diagnosis of infection. Different viral markers can be optimally detected at different stages of infection. Of particular clinical importance is the early identification of infection, which is pivotal for disease management and the development of blood screening assays. Non-structural protein 1 (NS1) is an early surrogate marker of infection and its detection in serum coincides with detectable viraemia. The aim of this work was to isolate and characterise serotype-specific monoclonal antibodies that bind to NS1 for each of the four DENV serotypes. This was achieved using phage display and a subtractive biopanning strategy to direct the antibody selection towards serotype-specific epitopes. This antibody isolation strategy has advantages over immunisation techniques where it is difficult to avoid antibody responses to cross-reactive, immunodominant epitopes. Serotype specificity to recombinant antigen for each of the antibodies was confirmed by Enzyme Linked Immunosorbent Assay (ELISA) and Surface Plasmon Resonance. Confirmation of binding to native DENV NS1 was achieved using ELISA and immunofluorescence assay on DENV infected Vero cells. No cross-reactivity with Zika or Kunjin viruses was observed. A previously isolated pan-reactive antibody that binds to an immunodominant epitope was able to pair with each of the serotype-specific antibodies in a sandwich ELISA, indicating that the serotype specific antibodies bind to epitopes which are all spatially distinct from the immunodominant epitope. These antibodies were suitable for use in a multiplexed assay for simultaneous detection and serotyping of DENV NS1 in human serum. This work demonstrates that phage display coupled with novel biopanning strategies is a valuable in vitro methodology for isolation of binders that can discern amongst
De Ridder, L; Maes, D; Dewulf, J; Butaye, P; Pasmans, F; Boyen, F; Haesebrouck, F; Van der Stede, Y
Salmonella enterica infection in pigs is economically important and poses a zoonotic risk. In this study, the efficacy of an attenuated S. enterica serovar Typhimurium strain was evaluated in three farrow-to-finish pig herds. In each herd, 120 piglets were vaccinated orally at 3 and 24 days of age, while 120 piglets served as unvaccinated controls. Faeces, ileocaecal lymph nodes and caecal contents were examined for S. Typhimurium by isolation and serum was analysed for antibodies against S. Typhimurium by ELISA. All pigs were weighed at pre-weaning and slaughter to determine daily weight gain. In vaccinated pigs prior to slaughter, significantly fewer animals excreted S. enterica, there was a significantly lower S. enterica-specific mean antibody titre and there was a significantly higher mean daily weight gain compared to unvaccinated controls. In two herds, there were significantly lower proportions of S. enterica positive ileocaecal lymph nodes and caecal contents at slaughter between the vaccinated and control groups, but this difference was not significant across all three herds. S. enterica with the same auxotrophic characteristics and genotype as the vaccine strain was isolated from several samples of faeces, ileocaecal lymph nodes and caecal contents from vaccinated pigs. These findings indicate that vaccination with an attenuated S. Typhimurium strain reduces S. enterica shedding, but the reduction is not consistent and the vaccine strain may persist in tissues. Copyright © 2014 Elsevier Ltd. All rights reserved.
Full Text Available 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.
Moore, Catrin E.; Sengduangphachanh, Amphone; Thaojaikong, Thaksinaporn; Sirisouk, Joy; Foster, Dona; Phetsouvanh, Rattanaphone; McGee, Lesley; Crook, Derrick W.; Newton, Paul N.; Peacock, Sharon J.
A prospective hospital-based study was undertaken to define the incidence of invasive pneumococcal disease (IPD) and circulating serotypes in Laos. Of 10,799 patients with hemocultures and 353 patients with cerebrospinal fluid samples, 0.21% and 5.4%, respectively, were positive for Streptococcus pneumoniae, giving a total of 35 IPD patients. We developed a real-time polymerase chain reaction to detect serotypes represented in the 13-valent pneumococcal vaccine. A blinded evaluation comparing serotype as defined by the Quellung reaction versus the polymerase chain reaction demonstrated 100% concordance. The most frequent serotype (n = 33 patients) was 1 (n = 6), followed by serotypes 5, 6A/B/C, 14, and 23F. Serotypes represented in the 7-valent polysaccharide-protein conjugate vaccine (PCV-7) infected 39% of patients, with 73% coverage for the PCV-10 and PCV-13 vaccines. Although the sample size is small, these data suggest that the PCV-7 vaccine may have relatively low efficacy in Laos. Further studies are urgently needed to guide pneumococcal vaccine policy in Laos. PMID:20810803
Hjálmarsdóttir, Martha Á; Gumundsdóttir, Pálína Fanney; Erlendsdóttir, Helga; Kristinsson, Karl G; Haraldsson, Gunnsteinn
Pneumococci are common colonizer, especially of children, and cocolonization of different serotypes is an important factor for intraspecies genetic exchange. The aim of this study was to analyze pneumococcal carriage and serotype distribution in unvaccinated healthy children in Iceland and compare conventional culture methods and molecular methods using DNA extracted directly from the samples. Nasopharyngeal swabs were obtained from 514 children aged 2-6 year attending day care centers in Reykjavik in 2009. The swabs were selectively cultured for pneumococci and the isolates serotyped using latex agglutination. DNA was also extracted directly from the swabs and serotyped using a multiplex PCR panel designed to detect vaccine serotypes and the most commonly carried non-vaccine serotypes. Pneumococcal carriage was detected in 391 (76.1%) of the children using polymerase chain reaction (PCR) and in 371 (72.2%) using conventional methods. Cocolonization was detected in 92 (23.5%) of the carriers when PCR method was used and in 30 (8.1%) when conventional methods were used, detecting 500 and 401 strains, respectively (P < 0.0001). The most common serotypes were 23F, 19A, 6B, 6A and 19F, rates 13-8%. The number of isolates of serotypes included in the 10-valent and 13-valent vaccines and detected by PCR were 234 (58.4%) and 363 (90.5%), respectively and by conventional methods 186 (46.4%) and 293 (73.1%), respectively. Cocolonization was detected in a fourth of the children carrying pneumococci using DNA extracted directly from nasopharyngeal swabs. The rate of carriage was very high, but no serotype dominated, and the children were commonly colonized by vaccine serotypes, especially cocolonized children.
Thrane, Sandra Wingaard; Taylor, Véronique L.; Lund, Ole
aeruginosa serotyper (PAst) program, which enabled in silico serotyping of P. aeruginosa isolates using WGS data. PAst has been made publically available as a web-service, and aptly facilitate high-throughput serotyping analysis. The program overcomes critical issues such as the loss of in vitro typeability...
Prins-van Ginkel, Annemarijn C|info:eu-repo/dai/nl/413970434; Berbers, Guy A M; Grundeken, Lucienne H; Tcherniaeva, Irina; Wittenberns, Jelle I; Elberse, Karin; Mollema, Liesbeth; de Melker, Hester E; Knol, Mirjam J
INTRODUCTION: Introduction of pneumococcal conjugate vaccines (PCVs) for infants decreased overall invasive pneumococcal disease (IPD), while non-vaccine serotype IPD increased. To fully understand this serotype replacement, knowledge about serotype dynamics in the pre-vaccine era is needed. In
Jessing, Stine Graakjær; Ahrens, Peter; Inzana, Thomas J.
serotypes. In serotypes 6 and 7 the sequenced DNA regions comprised five and four open reading frames, respectively, designated cps6ABCDE and cps 7ABCD, whereas the sequenced DNA region in serotype 12 comprised only two open reading frames designated cps12AB. At the amino acid level, CpsA, CpsB, and Cps...
Lay Ching Chai
Full Text Available 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 of typhoid 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.
Pornruangwong, Srirat; Hendriksen, Rene S; Pulsrikarn, Chaiwat; Bangstrakulnonth, Aroon; Mikoleit, Matthew; Davies, Rob H; Aarestrup, Frank M; Garcia-Migura, Lourdes
Salmonella enterica serovar Kedougou is among the top 10 serovars reported in northern Thailand. The objective of this study was to identify risk factors associated with Salmonella Kedougou infection in Thailand and to compare the molecular types and antimicrobial resistance with Salmonella Kedougou isolates of human origin from United States and of animal origin from the United Kingdom. Data from 13,976 Salmonella infections of which 253 were Salmonella Kedougou collected in Thailand between 2002 and 2008 were analyzed by logistic regression. Antimicrobial susceptibility testing and pulsed-field gel electrophoresis (PFGE) were performed on selected Salmonella Kedougou strains causing infections in Thailand (n = 66), and compared to isolates from the United States (n = 5) and the United Kingdom (n = 20). Logistic analysis revealed season (hot/dry; p = 0.023), region (northern Thailand; p Thailand were resistant to third-generation cephalosporins: two harbored bla(CTX-M-63) and one bla(CMY-2). PFGE revealed 45 unique clusters. Isolates obtained from humans in Thailand and the United States presented identical PFGE profiles suggesting a travel association, whereas the majority of the animal isolates from United Kingdom clustered separately. This study reveals Salmonella Kedougou as a major cause of human infections in northern Thailand especially during the hot period and suggests a global spread probably due to travel. The clonal types causing infections in humans differed from those observed in animals in United Kingdom, which suggests the absence of an epidemiological link and could suggest differences in virulence. The high frequency of antimicrobial resistance, including emergence of resistance to fluoroquinolones and third-generation cephalosporins, might pose problems for treatment of infections.
Salmonella isolated from chicken carcasses from a slaughterhouse in the state of Mato Grosso, Brazil: antibiotic resistance profile, serotyping, and characterization by repetitive sequence-based PCR system.
Cunha-Neto, Adelino da; Carvalho, Larrayane Albuês; Carvalho, Ricardo César Tavares; Dos Prazeres Rodrigues, Dália; Mano, Sergio Borges; Figueiredo, Eduardo Eustáquio de Souza; Conte-Junior, Carlos Adam
Salmonella is one of the major causative agents of foodborne infections. Salmonellosis becomes more dangerous when strains resistant to several antibiotics are found in food, especially in chicken, one of the primary transmission vehicles of this pathogen for humans. The present study aimed to estimate the occurrence of Salmonella in chicken carcasses from the state of Mato Grosso, Brazil, as well as determine the antibiotic resistance profile and genotypic characteristic of multi-drug resistant (MDR) isolates. During a 15-month period, from 01/2014 to 05/2015, 850 samples of chilled fresh chicken carcasses were sampled from a slaughterhouse and submitted to Salmonella determinations according to the ISO-6579/2002 method, serotyping and multiplex PCR. The disc diffusion test was applied for 17 antibiotics, according to CLSI (2014). Five isolates were genotyped by repetitive sequence-based PCR using the semi-automated DiversiLab (bioMérieux®) system. The occurrence of Salmonella in chicken carcasses was of 3.7% (31/850), with only 4 strains (12.9%) presenting as MDR, and 6 strains (19.35%) displaying ESBL. The predominant serovars were Salmonella Infantis (35.4%, 11/31), and S. Abony (25.8%, 8/31), followed by serovars S. Agona (12.9%, 4/31), S. Schwarzengrund (9.7%, 3/31), S. Anatum and Salmonella enterica O:4,5 (6.5%, 2/31), and only one Salmonella enterica O:6,7 strain (3.2%, 1/31). All isolates were resistant to one to 5 classes of antibiotics in decreasing order: folate pathway inhibitors, β-lactams (cephalosporins, penicillin, monobactams), tetracyclines, chloramphenicol, and gentamicin. However, strains sensitive to florfenicol, streptomycin, nalidixic acid, ciprofloxacin, enrofloxacin, and nitrofurantoin were also found in this study. Genotyping revealed 98 to 99% homology between 3 Salmonella strains, which displayed high phenotypic resistance similarity to β-lactams and folate pathway inhibitors. Detection of MDR non-typhoid Salmonella in chicken
Live attenuated Salmonella enterica serovar Choleraesuis vaccine vector displaying regulated delayed attenuation and regulated delayed antigen synthesis to confer protection against Streptococcus suis in mice.
Ji, Zhenying; Shang, Jing; Li, Yuan; Wang, Shifeng; Shi, Huoying
Salmonella enterica serotype Choleraesuis (S. Choleraesuis) and Streptococcus suis (S. suis) are important swine pathogens. Development of a safe and effective attenuated S. Choleraesuis vaccine vector would open a new window to prevent and control pig diseases. To achieve this goal, the mannose and arabinose regulated delayed attenuated systems (RDAS), Δpmi and ΔPcrp::TT araC PBADcrp, were introduced into the wild type S. Choleraesuis strain C78-3. We also introduced ΔrelA::araC PBADlacI TT to achieve regulated delayed antigen synthesis and ΔasdA to constitute a balanced-lethal plasmid system. The safety and immunogenicity of the resulted RDAS S. Choleraesuis strain rSC0011 carrying 6-phosphogluconate dehydrogenase (6-PGD) of S. suis serotype 2 (SS2) were evaluated in vitro and in vivo. Compared with the wild type parent strain C78-3 and vaccine strain C500, a live attenuated S. Choleraesuis vaccine licensed for piglet in China, the results showed that the survival curves of the vaccine strain rSC0011 were similar to those of strains C78-3 and C500 at the early stage of infection, but lower than those of C78-3 and higher than those of C500 at the later stage in both porcine alveolar macrophages and peripheral porcine monocytes. The LD50 of the RDAS strains rSC0011 by oral route in mice was close to that of C500 and 10,000-fold higher than that of C78-3. Similar results were achieved by intraperitoneal (i.p.) route, suggesting that the RDAS strains rSC0011 achieved similar attenuation as C500. However, the RDAS strain rSC0011 was superior to C500 in colonization of Peyer's patches. Adult mice orally immunized with strain rSC0011 carrying a plasmid expression 6-phosphogluconate dehydrogenase (6-PGD) gene from SS2 developed strong immune responses against 6-PGD and Salmonella antigens, and conferred high protection against i.p. challenge with SS2. Copyright © 2015 Elsevier Ltd. All rights reserved.
Lamas, A; Fernandez-No, I C; Miranda, J M; Vázquez, B; Cepeda, A; Franco, C M
Salmonella serovars are responsible for foodborne diseases around the world. The ability to form biofilms allows microorganisms to survive in the environment. In this study, 73 Salmonella strains, belonging to four different subspecies, were isolated from poultry houses and foodstuffs and tested. Biofilm formation was measured at four different temperatures and two nutrient concentrations. Morphotypes and cellulose production were evaluated at three different temperatures. The presence of several genes related to biofilm production was also examined. All strains and subspecies of Salmonella had the ability to form biofilms, and 46.57% of strains produced biofilms under all conditions tested. Biofilm formation was strain dependent and varied according to the conditions. This is the first study to analyze biofilm formation in a wide number of Salmonella enterica subsp. arizonae strains, and no direct relationship between the high prevalence of Salmonella enterica subsp. arizonae strains and their ability to form biofilm was established. Morphotypes and cellulose production varied as the temperature changed, with 20°C being the optimum temperature for expression of the red, dry, and rough morphotype and cellulose. Salmonella enterica subsp. arizonae, whose morphotype is poorly studied, only showed a smooth and white morphotype and lacked the csgD and gcpA genes that are implicated in biofilm production. Thus, Salmonella biofilm formation under different environmental conditions is a public health problem because it can survive and advance through the food chain to reach the consumer.
Cartas Espinel, Irene; Guerra, Priscila Regina; Jelsbak, Lotte
Polyamines (putrescine and spermidine) are small-cationic amines ubiquitous in nature and present in most living cells. In recent years they have been linked to virulence of several human pathogens including Shigella spp and Salmonella enterica serovar Typhimurium (S. Typhimurium). Central to S...
Nontyphoidal Salmonella strains are the main source of pathogenic bacterial contamination in the poultry industry. Recently, Salmonella enterica serovar Kentucky has been recognized as the most prominent serovar on carcasses in poultry-processing plants. Previous studies showed that flagella are one...
Valls Serón, M; Haiko, J; DE Groot, P G; Korhonen, T K; Meijers, J C M
Pathogenic bacteria modulate the host coagulation system to evade immune responses or to facilitate dissemination through extravascular tissues. In particular, the important bacterial pathogens Salmonella enterica and Yersinia pestis intervene with the plasminogen/fibrinolytic system. Thrombin-activatable fibrinolysis inhibitor (TAFI) has anti-fibrinolytic properties as the active enzyme (TAFIa) removes C-terminal lysine residues from fibrin, thereby attenuating accelerated plasmin formation. Here, we demonstrate inactivation and cleavage of TAFI by homologous surface proteases, the omptins Pla of Y. pestis and PgtE of S. enterica. We show that omptin-expressing bacteria decrease TAFI activatability by thrombin-thrombomodulin and that the anti-fibrinolytic potential of TAFIa was reduced by recombinant Escherichia coli expressing Pla or PgtE. The functional impairment resulted from C-terminal cleavage of TAFI by the omptins. Our results indicate that TAFI is degraded directly by the omptins PgtE of S. enterica and Pla of Y. pestis. This may contribute to the ability of PgtE and Pla to damage tissue barriers, such as fibrin, and thereby to enhance spread of S. enterica and Y. pestis during infection. © 2010 International Society on Thrombosis and Haemostasis.
Hendriksen, Rene S.; Joensen, Katrine Grimstrup; Lukwesa-Musyani, Chileshe
Two cases of extremely drug-resistant Salmonella enterica serovar Senftenberg isolated from patients in Zambia were investigated by utilizing MIC determinations and whole-genome sequencing. The isolates were resistant to, and harbored genes toward, nine drug classes, including fluoroquinolones an...... and extended-spectrum cephalosporins, contained two plasmid replicons, and differed by 93 single-nucleotide polymorphisms....
Kutter, Stefan; Hartmann, Anton; Schmid, Michael
Colonization of barley plants by the food-borne pathogens Salmonella enterica serovar typhimurium and three Listeria spp. (L. monocytogenes, L. ivanovii, L. innocua) was investigated in a monoxenic system. Herbaspirillum sp. N3 was used as a positive control and Escherichia coli HB101 as a negative control for endophytic root colonization. Colonization of the plants was tested 1-4 weeks after inoculation by determination of CFU, specific PCR assays and fluorescence in situ hybridization (FISH) with fluorescently labelled oligonucleotide probes in combination with confocal laser scanning microscopy (CLSM). Both S. enterica strains were found as endophytic colonizers of barley roots and reached up to 2.3 x 10(6) CFU per g root fresh weight after surface sterilization. The three Listeria strains had 10-fold fewer cell numbers after surface sterilization on the roots and therefore were similar to the results of nonendophytic colonizers, such as E. coli HB101. The FISH/CSLM approach demonstrated not only high-density colonization of the root hairs and the root surface by S. enterica but also a spreading to subjacent rhizodermis layers and the inner root cortex. By contrast, the inoculated Listeria spp. colonized the root hair zone but did not colonize other parts of the root surface. Endophytic colonization of Listeria spp. was not observed. Finally, a systemic spreading of S. enterica to the plant shoot (stems and leaves) was demonstrated using a specific PCR analysis and plate count technique.
Sandvang, Dorthe; Aarestrup, Frank Møller; Jensen, Lars Bogø
The presence and genetic content of integrons was investigated in eight Salmonella enterica Typhimurium DT104 isolates from different pig herds in Denmark. Two different integrons were identified using PCR and sequencing. Each of the integrons carried a single resistance cassette in addition to t...
Multidrug-resistant (MDR) Salmonella is associated with increased morbidity in humans and presents an important food safety concern. Antibiotic resistance among isolates of Salmonella enterica serovar Typhimurium has become especially prevalent as over 27 per cent of isolates from humans in the Unit...
Dasgupta, Asish; Mukherjee, Sayanti; Chaki, Shaswati
When administered to mice at doses of 100microg/mouse and 200microg/mouse, thioridazine (TDZ) significantly protected animals from the lethality produced by a virulent strain of Salmonella enterica serovar Typhimurium and reduced the number of bacteria retrieved from the spleen, liver and heart...
Hatta, Mochammad; Sultan, Andi R.; Pastoor, Rob; Smits, Henk L.
Phase variation is a property unique of some Salmonella enterica serovar Typhi strains from Indonesia. Salmonella Typhi isolates from Indonesia have been described that in addition to the phase 1 Hd flagellin gene contain a second flagellin gene named z66. S. Typhi isolates from Indonesia with a
Leekitcharoenphon, Pimlapas; Nielsen, Eva M.; Kaas, Rolf Sommer
Salmonella enterica is a common cause of minor and large food borne outbreaks. To achieve successful and nearly ‘real-time’ monitoring and identification of outbreaks, reliable sub-typing is essential. Whole genome sequencing (WGS) shows great promises for using as a routine epidemiological typing...
The objectives of the present study were to investigate the effectiveness of zein-based coatings in reducing populations of Salmonella enterica serovar Typhimurium and preserving quality of cherry tomatoes. Tomatoes were inoculated with a cocktail of S. Typhimurium LT2 plus three mutants on the smoo...
Rodicio, M. Rosario; Guerra, Beatriz; Hopkins, Katie L.
In developing countries, Salmonella enterica serovar Enteritidis causes substantial illness and death, and drug resistance is increasing. Isolates from the United Kingdom containing virulence-resistance plasmids were characterized. They mainly caused invasive infections in adults linked to Africa. The common features in isolates from these continents indicate the role of human travel in their spread. PMID:22709653
Hald, Birthe; Olsen, A.; Madsen, Mogens
In December 1994, Salmonella enterica serovar Infantis (S. Infantis) was accidentally introduced into a Danish broiler house by stocking an S. Infantis-infected broiler nock of 39,900 day-old chicks. At the time of the study, the infection had persisted through e broiler cycles. Typhaea stercorea...
Salmonella enterica, a human enteric pathogen, has the ability to multiply and survive endophytically in plants, and mutations in genes encoding the type III secretion system (T3SS) or its effectors (T3Es) may contribute to this colonization. Two reporter plasmids for T3E translocation into plant ce...
Hopkins, K.L.; Kirchner, M.; Guerra, B.; Granier, A.; Lucarelli, C.; Porrero, M.C.; Jakubczak, A.; Threlfall, J.; Mevius, D.J.
A marked increase in the prevalence of S. enterica serovar 4,,12:i:- with resistance to ampicillin, streptomycin, sulphonamides and tetracyclines (R-type ASSuT) has been noted in food-borne infections and in pigs/pig meat in several European countries in the last ten years. One hundred and
Iqbal, Muhammad Kashif; Ijaz, Muhammad; Aslam, Hassaan Bin; Farooqi, Shahid Hussain; Ahmad, Syed Saleem; Akhtar, Raheela
Salmonella enterica (S. enterica) is the major zoonotic threat for small ruminants and humans responsible for huge economic losses and high mortality in Pakistan. Lambs and kids of Lahore district were examined to determine the prevalence, hematology and chemotherapy of S. enterica. A total of 200 diarrheic samples (n=100 lambs; n=100 kids) were collected and examined; 59 (29.50%) were found positive for S. enterica. Lambs had lightly greater prevalence (31%) than kids (29%). The frequency analysis (OR=1.16 [reciprocal =0.87]) showed non-significant difference in both the lambs and kids. The significant decrease (Pℜ0.001) in hemoglobin, pack cell volume and total erythrocyte count was observed in infected lambs and kids. Results of in-vitro antibiotic susceptibility test revealed that S. enteric isolated from both lambs and kids were susceptible to levofloxacin, ciprofloxacin, ofloxacin, gentamicin, azithromycin, tobramycin, amoxicillin, ampicillin and nalidixic acid. Where as the results of in vivo antibiotic trials showed that isolates from both lambs and the kids with diarrhea were susceptible to levofloxacin and ciprofloxacin.
Full Text Available 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.
Condell, Orla; Iversen, Carol; Cooney, Shane; Power, Karen A; Walsh, Ciara; Burgess, Catherine; Fanning, Séamus
Biocides play an essential role in limiting the spread of infectious disease. The food industry is dependent on these agents, and their increasing use is a matter for concern. Specifically, the emergence of bacteria demonstrating increased tolerance to biocides, coupled with the potential for the development of a phenotype of cross-resistance to clinically important antimicrobial compounds, needs to be assessed. In this study, we investigated the tolerance of a collection of susceptible and multidrug-resistant (MDR) Salmonella enterica strains to a panel of seven commercially available food-grade biocide formulations. We explored their abilities to adapt to these formulations and their active biocidal agents, i.e., triclosan, chlorhexidine, hydrogen peroxide, and benzalkonium chloride, after sequential rounds of in vitro selection. Finally, cross-tolerance of different categories of biocidal formulations, their active agents, and the potential for coselection of resistance to clinically important antibiotics were investigated. Six of seven food-grade biocide formulations were bactericidal at their recommended working concentrations. All showed a reduced activity against both surface-dried and biofilm cultures. A stable phenotype of tolerance to biocide formulations could not be selected. Upon exposure of Salmonella strains to an active biocidal compound, a high-level of tolerance was selected for a number of Salmonella serotypes. No cross-tolerance to the different biocidal agents or food-grade biocide formulations was observed. Most tolerant isolates displayed changes in their patterns of susceptibility to antimicrobial compounds. Food industry biocides are effective against planktonic Salmonella. When exposed to sublethal concentrations of individual active biocidal agents, tolerant isolates may emerge. This emergence was associated with changes in antimicrobial susceptibilities.
Full Text Available The role of the Salmonella Pathogenicity Islands (SPIs in pathogenesis of Salmonella enterica Typhimurium infection in the chicken is poorly studied, while many studies have been completed in murine models. The Type VI Secretion System (T6SS is a recently described protein secretion system in Gram-negative bacteria. The genus Salmonella contains five phylogenetically distinct T6SS encoded in differentially distributed genomic islands. S. Typhimurium harbors a T6SS encoded in SPI-6 (T6SSSPI-6, which contributes to the ability of Salmonella to colonize mice. On the other hand, serotype Gallinarum harbors a T6SS encoded in SPI-19 (T6SSSPI-19 that is required for colonization of chicks. In this work, we investigated the role of T6SSSPI-6 in infection of chicks by S. Typhimurium. Oral infection of White Leghorn chicks showed that a ΔT6SSSPI-6 mutant had reduced colonization of the gut and internal organs, compared with the wild-type strain. Transfer of the intact T6SSSPI-6 gene cluster into the T6SS mutant restored bacterial colonization. In addition, our results showed that transfer of T6SSSPI-19 from S. Gallinarum to the ΔT6SSSPI-6 mutant of S. Typhimurium not only complemented the colonization defect but also resulted in a transient increase in the colonization of the cecum and ileum of chicks at days 1 and 3 post-infection. Our data indicates that T6SSSPI-6 contributes to chicken colonization and suggests that both T6SSSPI-6 and T6SSSPI-19 perform similar functions in vivo despite belonging to different phylogenetic families.
Tamamura, Yukino; Uchida, Ikuo; Tanaka, Kiyoshi; Okazaki, Hizuru; Tezuka, Satoru; Hanyu, Hideki; Kataoka, Natsumi; Makino, Sou-Ichi; Kishima, Masato; Kubota, Takayuki; Kanno, Toru; Hatama, Shinichi; Ishihara, Ryoko; Hata, Eiji; Yamada, Hironari; Nakaoka, Yuuji; Akiba, Masato
The molecular epidemiology of 545 Salmonella enterica serovar Typhimurium isolates collected between 1977 and 2009 from cattle in Hokkaido, Japan, was investigated using pulsed-field gel electrophoresis (PFGE). Nine main clusters were identified from 116 PFGE patterns. Cluster I comprised 248 isolates, 243 of which possessed a sequence specific to definitive phage type 104 (DT104) or U302. The cluster I isolates were dominant in 1993 to 2003, but their numbers declined beginning in 2004. Beginning in 2002, an increase was observed in the number of cluster VII isolates, consisting of 21 PFGE patterns comprising 165 isolates. A total of 116 isolates representative of the 116 PFGE profiles were analyzed by multilocus variable-number tandem-repeat analysis (MLVA). Other than two drug-sensitive isolates, 19 isolates within cluster VII were classified in the same cluster by MLVA. Among the cluster VII isolates, an antibiotic resistance type showing resistance to ampicillin, chloramphenicol, streptomycin, sulfonamides, tetracycline, kanamycin, cefazolin, and sulfamethoxazole-trimethoprim and a resistance type showing resistance to ampicillin, streptomycin, sulfonamides, tetracycline, and kanamycin were found in 23 and 125 isolates, respectively. In the 19 isolates representative of cluster VII, the bla(TEM-1) gene was found on a Salmonella serotype Typhimurium virulence plasmid, which was transferred to Escherichia coli by electroporation along with resistance to two to four other antimicrobials. Genomic analysis by subtractive hybridization and plasmid analysis suggested that the bla(TEM-1)-carrying virulence plasmid has a mosaic structure composed of elements of different origin. These results indicate an emerging multidrug-resistant S. Typhimurium clone carrying a virulence-resistance plasmid among cattle in Hokkaido, Japan.
Tamamura, Yukino; Uchida, Ikuo; Tanaka, Kiyoshi; Okazaki, Hizuru; Tezuka, Satoru; Hanyu, Hideki; Kataoka, Natsumi; Makino, Sou-ichi; Kishima, Masato; Kubota, Takayuki; Kanno, Toru; Hatama, Shinichi; Ishihara, Ryoko; Hata, Eiji; Yamada, Hironari; Nakaoka, Yuuji; Akiba, Masato
The molecular epidemiology of 545 Salmonella enterica serovar Typhimurium isolates collected between 1977 and 2009 from cattle in Hokkaido, Japan, was investigated using pulsed-field gel electrophoresis (PFGE). Nine main clusters were identified from 116 PFGE patterns. Cluster I comprised 248 isolates, 243 of which possessed a sequence specific to definitive phage type 104 (DT104) or U302. The cluster I isolates were dominant in 1993 to 2003, but their numbers declined beginning in 2004. Beginning in 2002, an increase was observed in the number of cluster VII isolates, consisting of 21 PFGE patterns comprising 165 isolates. A total of 116 isolates representative of the 116 PFGE profiles were analyzed by multilocus variable-number tandem-repeat analysis (MLVA). Other than two drug-sensitive isolates, 19 isolates within cluster VII were classified in the same cluster by MLVA. Among the cluster VII isolates, an antibiotic resistance type showing resistance to ampicillin, chloramphenicol, streptomycin, sulfonamides, tetracycline, kanamycin, cefazolin, and sulfamethoxazole-trimethoprim and a resistance type showing resistance to ampicillin, streptomycin, sulfonamides, tetracycline, and kanamycin were found in 23 and 125 isolates, respectively. In the 19 isolates representative of cluster VII, the blaTEM-1 gene was found on a Salmonella serotype Typhimurium virulence plasmid, which was transferred to Escherichia coli by electroporation along with resistance to two to four other antimicrobials. Genomic analysis by subtractive hybridization and plasmid analysis suggested that the blaTEM-1-carrying virulence plasmid has a mosaic structure composed of elements of different origin. These results indicate an emerging multidrug-resistant S. Typhimurium clone carrying a virulence-resistance plasmid among cattle in Hokkaido, Japan. PMID:21239560
Han, Jing; Pendleton, Sean J; Deck, Joanna; Singh, Ruby; Gilbert, Jeffrey; Johnson, Timothy J; Sanad, Yasser M; Nayak, Rajesh; Foley, Steven L
Antimicrobial resistance in Salmonella enterica is often plasmid encoded. A key resistance plasmid group is the incompatibility group (Inc) A/C plasmids that often carry multiple resistance determinants. Previous studies showed that IncA/C plasmids were often co-located with other plasmids. The current study was undertaken to evaluate the impact of plasmid co-carriage on antimicrobial resistance and plasmid transfer. A total of 1267 Salmonella isolates, representing multiple serotypes and sources were previously subjected to susceptibility testing and 251 isolates with resistance to at least 5 antimicrobial agents were identified for further study. Each isolate was subjected to PCR-based replicon typing, and those with IncA/C plasmids were selected for plasmid isolation, PCR-based mapping of IncA/C plasmid backbone genes, and conjugation assays to evaluate resistance plasmid transferability. Of the 87 identified IncA/C positive isolates, approximately 75% carried a plasmid with another identified replicon type, with the most common being I1 (39%), FIA, FIIA, FIB and HI2 (each 15%). PCR-based mapping indicated significant diversity in IncA/C backbone content, especially in regions encoding transfer-associated and hypothetical proteins. Conjugation experiments showed that nearly 68% of the isolates transferred resistance plasmids, with 90% containing additional identified plasmids or larger (>50 kb) non-typeable plasmids. The majority of IncA/C-positive strains were able to conjugally transfer antimicrobial resistance to the recipient, encoded by IncA/C and/or co-carried plasmids. These findings highlight the importance of co-located plasmids for resistance dissemination either by directly transferring resistance genes or by potentially providing the needed conjugation machinery for IncA/C plasmid transfer. Copyright © 2018. Published by Elsevier B.V.
Serovariedades y patrones de susceptibilidad a los antimicrobianos de cepas de Salmonella aisladas de alimentos en Cuba Serotypes and antimicrobial susceptibility patterns of Salmonella strains isolated from food in Cuba
Yamila Puig Peña
Full Text Available Se describen los serotipos de 178 cepas de Salmonella enterica aisladas de alimentos en diferentes regiones de Cuba entre enero de 2008 y diciembre de 2009, y el patrón de susceptibilidad a los antimicrobianos de 100 aislados seleccionados mediante muestreo por estratos. Se identificaron 20 serovariedades de Salmonella entre las que predominaron S. Enteritidis (23%; S. Agona (13,5% y S. London (11,2%. Del total, 75% de las cepas fueron resistentes o presentaron resistencia intermedia a al menos uno de los fármacos probados, en el siguiente orden, según su frecuencia: tetraciclina (70,7%; ampicilina (22,7% y ácido nalidíxico (14,7%. Se identificaron 10 patrones de resistencia diferentes y predominaron las cepas resistentes o conresistencia intermedia a un fármaco (89,3%. Tres cepas (S. Infantis, S. Derby y S. Enteritidis fueron multirresistentes y una, de S. Enteritidis, dio un resultado no sensible al ácido nalidíxico y la ciprofloxacina. Se destaca la necesidad de extremar la vigilancia sanitaria integrada en el país para el control de la salmonelosis.The serotypes of 178 isolates of Salmonella enterica taken from food in different regions of Cuba between January 2008 and December 2009 were identified, and the antimicrobial susceptibility pattern of 100 selected isolates was determined by strata sampling. A total of 20 Salmonella serotypes were identified, with a predominance of S. Enteritidis (23%, S. Agona (13.5%, and S. London (11.2%. Of all the strains, 75% were resistant or presented intermediate resistance to at least one of the drugs tested, in the following order: tetracycline (70.7%, ampicillin (22.7%, and nalidixic acid (14.7%. Ten different resistance patterns were identified. The most frequent patterns corresponded to strains that were either drugresistant or had intermediate resistance (89.3%. Three strains (identified as S. Infantis, S. Derby, and S. Enteritidis were multiresistant, and one of them, S. Enteritidis, was
Fett, William F
Foodborne illness due to the consumption of contaminated raw or lightly cooked sprouts is a continuing food safety concern. In this study, we tested several plant-associated pseudomonads for their ability to inhibit the growth of Salmonella enterica both in vitro and in situ. An agar spot bioassay method was used with three different media. Only Pseudomonas fluorescens 2-79 produced clear zones of inhibition when tested against five serovars of S. enterica, and activity was dependent on media type and serovar. The antibiosis by derivative strains of P. fluorescens 2-79 defective in the production of phenazine-1-carboxylic acid and fluorescent siderophore was not reduced, indicating that these known antimicrobial metabolites were not responsible for the inhibition observed in our studies. However, mutants defective in the regulatory gene gacS (global antibiotic and cyanide control) were severely reduced in inhibitory activity. In tryptic soy broth, the control cultures of a cocktail of S. enterica strains reached approximately 10 log CFU/ml by 24 h but, when coinoculated with P. fluorescens 2-79, reached only approximately 5 log CFU/ml. The addition of P. fluorescens 2-79 to the seed soak water prior to the germination of alfalfa seed previously inoculated with a cocktail of S. enterica strains led to an average reduction of 5 log CFU/g at 6 days of sprouting without an adverse effect on sprout yield or appearance. Time course studies indicated that S. enterica outgrowth was controlled on days 1 through 6 of sprouting. Competitive exclusion as a potential food safety intervention for seed sprouts merits further study.
Muthuirulandi Sethuvel, D P; Devanga Ragupathi, N K; Anandan, S; Veeraraghavan, B
Shigellosis represents a major burden of disease in developing countries. A low infectious dose allows the disease to be spread effectively. Although shigellosis is mostly a self-limiting disease, antibiotics are recommended to reduce deaths, disease symptoms and organism-shedding time. However, in India, antimicrobial resistance among the genus Shigella is more common than among any other enteric bacteria. Notably, new serotypes or subserotypes in Shigella are reported from various parts of the world. Identification of new subserotypes of Shigella spp. is becoming a major issue as these strains are nontypeable by conventional serotyping. The commercially available antisera may not cover all possible epitopes of the O lipopolysaccharide antigen of Shigella serotypes. Therefore, molecular methods which most closely approach the resolution of full serotyping are necessary to identify such strains. In addition, the knowledge of a prevalent serotype in various geographic regions may assist in formulating strategies such as the development of a vaccine to prevent infection especially when the immunity to disease is serotype specific, and to understand the disease burden caused by new Shigella serotypes. © 2016 The Society for Applied Microbiology.
Huang, Xiaodong; Clements, Archie C. A.; Williams, Gail; Devine, Gregor; Tong, Shilu; Hu, Wenbiao
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.
Full Text Available Objective: To identify the homology and phylogenetic relationship among the four dengue virus (DENV serotypes, and conservation of amino acid in E-proteins and to find out the phylogenetic relationship among the strains of four DENV serotypes. Methods: Clustal W analysis for homology and phylogram, European molecular biology open software suite for pairwise alignment of amino acid sequences and BLAST-P analysis for various strains of four DENV serotypes were carried out. Results: Homology of E-protein sequences of four DENV serotypes indicated a close relationship of DENV-1 with DENV-3. DENV-2 showed close relationship with DENV-1 and -3 forming a single cluster whereas DENV-4 alone formed group with a single serotype. In the multiple sequence alignment, 19 amino acid conserved groups were observed. BLAST-P analysis showed more number of 100% similarity among DENV-1 and -3 strains whereas only few strains showed 100% similarity in DENV-4. However, 100% similarity was absent among the DENV-3 strains. Conclusions: From the present study, phylogenetically all the four DENV serotypes were related but DENV-1, -2 and -3 were very closely related whereas DENV-4 was somewhat distant from the other three serotypes.
Nandi, S P; Rahman, M Z; Momtaz, S; Sultana, M; Hossain, M A
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. © 2013 Blackwell Verlag GmbH.
We need to raise the issue that focus on children as the only carriage group for pneumococci is not optimal; we need to consider that other age groups might also be carriers of pneumococcal serotypes causing invasive pneumococcal diseases (IPD) in unvaccinated age groups. The pneumococcal conjugate vaccines (PCV) have successfully removed IPD from vaccinated children. Studies have shown an effect of PCV reducing the pneumococcal carriage of PCV serotypes in children. The status for several countries having used PCV for many years is that they do not see PCV serotypes neither carried nor as a cause of IPD in children. PCV vaccination of children has shown a herd protection effect in unvaccinated groups as a reduction in IPD cases caused by PCV serotypes. However, not all PCV serotypes have disappeared as the cause of IPD in the unvaccinated age groups. The author therefore believes that if we are to see PCV serotypes disappear as a cause of IPD in unvaccinated age groups, we need to perform further carriage studies to examine carriage in other age groups. Alternatively, all age groups should be vaccinated against pneumococci to eliminate IPD caused by PCV serotypes from possible hidden carriers.
The 7-valent pneumococcal conjugate vaccine (PCV7) was included in the routine infant immunization schedule in Ireland in September 2008. We determined the serotype of 977 S. pneumoniae isolates causing invasive disease between 2000-2002 and 2007-2008, assessed for the presence of the recently described serotype 6C and determined the susceptibility of isolates during 2007-2008 to penicillin and cefotaxime. Serotype 14 was the most common serotype during both periods and 7·7% of isolates previously typed as serotype 6A were serotype 6C. During 2000-2002 and 2007-2008, PCV7 could potentially have prevented 85% and 74% of invasive pneumococcal disease in the target population (i.e. children aged <2 years), respectively. The level of penicillin non-susceptibility was 17% in 2007-2008. Ongoing surveillance of serotypes is required to determine the impact of PCV7 in the Irish population and to assess the potential of new vaccines with expanded valency.
Full Text Available An observational study was conducted to know the serotypes and antimicrobial susceptibility of isolates of Streptococcus pneumoniae responsible for meningitis in Cuba, where there is no vaccine yet to prevent invasive pneumococcal disease. The study included the total number of isolates submitted to the "Pedro Kourí" Institute between 2007 and 2012 (N=237. Serotypes identification was performed using capsular swelling test and antimicrobial susceptibility was studied by determining the minimum inhibitory concentration using the broth microdilution method. Predominant serotypes were 6A, 6B, 14, 19F and 23F and other non-vaccinal 18 serogroups/serotypes were identified in 29.1% of the isolates. A tendency to an increased resistance to penicillin (44.3 % was observed; the most common resistance patterns were: penicillin-trimethoprim/sulfamethoxazole and penicillin-erythromycin (21.1% and 10.5%, respectively. The largest number of isolates resistant to penicillin was in serotypes 6B, 14, 19F and 23F and the possibility of resistant non-vaccine serotypes emergence should be considered. The results show that 70.4 % of the isolates studied corresponds to the serotypes included in 13-valent conjugated pneumococcal vaccine, but with 10-valent it would achieve a lower vaccination potential coverage (56.1%. This information must be considered when evaluating the decision to use in Cuba any commercially available vaccine or the proposal of another strategy of vaccination from autochthonous vaccine candidates.
Sunil K Khattar
Full Text Available The nine serotypes of avian paramyxoviruses (APMVs are frequently isolated from domestic and wild birds worldwide. APMV-1, also called Newcastle disease virus, was shown to be attenuated in non-avian species and is being developed as a potential vector for human vaccines. In the present study, we extended this evaluation to the other eight serotypes by evaluating infection in BALB/c mice. Mice were inoculated intranasally with a prototype strain of each of the nine serotypes and monitored for clinical disease, gross pathology, histopathology, virus replication and viral antigen distribution, and seroconversion. On the basis of multiple criteria, each of the APMV serotypes except serotype 5 was found to replicate in mice. Five of the serotypes produced clinical disease and significant weight loss in the following order of severity: 1, 2>6, 9>7. However, disease was short-lived. The other serotypes produced no evident clinical disease. Replication of all of the APMVs except APMV-5 in the nasal turbinates and lungs was confirmed by the recovery of infectious virus and by substantial expression of viral antigen in the epithelial lining detected by immunohistochemistry. Trace levels of infectious APMV-4 and -9 were detected in the brain of some animals; otherwise, no virus was detected in the brain, small intestine, kidney, or spleen. Histologically, infection with the APMVs resulted in lung lesions consistent with broncho-interstitial pneumonia of varying severity that were completely resolved at 14 days post infection. All of the mice infected with the APMVs except APMV-5 produced serotype-specific HI serum antibodies, confirming a lack of replication of APMV-5. Taken together, these results demonstrate that all APMV serotypes except APMV-5 are capable of replicating in mice with minimal disease and pathology.
Phenotype microarrays were analyzed for 51 datasets derived from Salmonella enterica. The top 4 serovars associated with poultry products and one associated with turkey, respectively Typhimurium, Enteritidis, Heidelberg, Infantis and Senftenberg, were represented. Datasets were clustered into two ...
Kajikawa, A.; Satoh, E.; Leer, R.J.; Yamamoto, S.; Igimi, S.
A recombinant Lactobacillus casei expressing a flagellar antigen from Salmonella enterica serovar Enteritidis was constructed and evaluated as a mucosal vaccine. Intragastric immunization of the recombinant strain conferred protective immunity against Salmonella infection in mice. This immunization
A Constitutively Mannose-Sensitive Agglutinating Salmonella enterica subsp. enterica Serovar Typhimurium Strain, Carrying a Transposon in the Fimbrial Usher Gene stbC, Exhibits Multidrug Resistance and Flagellated Phenotypes
Full Text Available Static broth culture favors Salmonella enterica subsp. enterica serovar Typhimurium to produce type 1 fimbriae, while solid agar inhibits its expression. A transposon inserted in stbC, which would encode an usher for Stb fimbriae of a non-flagellar Salmonella enterica subsp. enterica serovar Typhimurium LB5010 strain, conferred it to agglutinate yeast cells on both cultures. RT-PCR revealed that the expression of the fimbrial subunit gene fimA, and fimZ, a regulatory gene of fimA, were both increased in the stbC mutant when grown on LB agar; fimW, a repressor gene of fimA, exhibited lower expression. Flagella were observed in the stbC mutant and this phenotype was correlated with the motile phenotype. Microarray data and RT-PCR indicated that the expression of three genes, motA, motB, and cheM, was enhanced in the stbC mutant. The stbC mutant was resistant to several antibiotics, consistent with the finding that expression of yhcQ and ramA was enhanced. A complementation test revealed that transforming a recombinant plasmid possessing the stbC restored the mannose-sensitive agglutination phenotype to the stbC mutant much as that in the parental Salmonella enterica subsp. enterica serovar Typhimurium LB5010 strain, indicating the possibility of an interplay of different fimbrial systems in coordinating their expression.
Bruun, B; Gahrn-Hansen, B; Westh, H
Surveillance performed after the introduction of general Haemophilus influenzae serotype b (Hib) vaccination in Denmark identified 13 cases of invasive bacteraemic H. influenzae serotype f (Hif) disease in adults over a period of 7 years. Bacteraemic respiratory tract infections accounted for 61...... sequences. Multilocus enzyme electrophoresis typing revealed that recent Danish and American isolates belonged to a single Hif clone, which may be undergoing expansion. The need for accurate serotyping of H. influenzae to enable reliable monitoring for Hib replacement by other capsular types is emphasized....
Lock, J. L.; Board, R. G.
A study was made of the persistence of different Salmonella serotypes in hens' egg albumin in vitro at 4, 20 and 30 degrees C. The majority of serotypes remained viable but did not increase in numbers at 20 and 30 degrees C for 42 days. At 4 degrees C many of the serotypes died out. The addition of ferric ammonium citrate on the 42nd day of incubation induced multiplication of organisms incubated at 20 and 30 degrees C, but not at 4 degrees C. The pH and glucose concentration of the albumen d...
Notermans, S; Chakraborty, T; Leimeister-Wächter, M; Dufrenne, J; Heuvelman, K J; Maas, H; Jansen, W; Wernars, K; Guinee, P
A total of 284 strains of Listeria, including all known serovars and biovars together with Listeria grayi and Listeria murrayi, were biotyped and serotyped. Biotyping and serotyping could be done in 2 days. A gene probe encoding a delayed hypersensitivity factor (DTH) was used in the detection of pathogenic biotypes and serotypes of the tested strains. The gene was found in all 117 tested Listeria monocytogenes strains of serogroups 1/2a, 1/2b, 1/2c, 3a, 3b, 3c, 4c, 4d, 4e, 4ab, and 7. It was...
Dekker, Niels; Daemen, Ineke; Verstappen, K.M.; Greeff, de A.; Smith, H.E.; Duim, Birgitta
Abstract Invasive Streptococcus suis (S. suis) infections in pigs are often associated with serotypes 2 and 9. Mucosal sites of healthy pigs can be colonized with these serotypes, often multiple serotypes per pig. To unravel the contribution of these serotypes in pathogenesis and epidemiology,
Parsons, Sandra K.; Bull, C. Michael
The life cycles of many enteric bacterial species require a transition between two very distinct environments. Their primary habitat is the gastrointestinal tract of the host, while their secondary habitat, during transmission from one host to another, consists of environments external to the host, such as soil, water, and sediments. Consequently, both host and environmental factors shape the genetic structure of enteric bacterial populations. This study examined the distribution of four Salmonella enterica subspecies in a population of sleepy lizards, Tiliqua rugosa, in a semiarid region of South Australia. The lizards living within the 1,920-m by 720-m study site were radio tracked, and their enteric bacteria were sampled at regular intervals throughout their active seasons in the years 2001, 2002, and 2006. Four of the six subspecies of S. enterica were present in this population and were nonrandomly distributed among the lizards. In particular, S. enterica subsp. diarizonae was restricted to lizards living in the most shaded parts of the study site with an overstorey of Casuarina trees. Experiments undertaken to investigate the survival of S. enterica cells under seminatural conditions revealed that cell survival decreased with increased exposure to elevated temperatures and UV light. Among the three S. enterica subspecies tested, S. enterica subsp. diarizonae consistently had an average expected life span that was shorter than that observed for the other two subspecies. There was no indication in the data that there was any competitive dominance hierarchy among the S. enterica subspecies within individual hosts. Thus, the nonrandom distribution of S. enterica subspecies in this population of lizards appears to be driven by their different survival characteristics in the external environment. PMID:26092451
Kim, Hyeun Bum; Singer, Randall S; Borewicz, Klaudyna; White, Bryan A; Sreevatsan, Srinand; Johnson, Timothy J; Espejo, Luis A; Isaacson, Richard E
To evaluate the effects of tylosin on C-reactive protein concentration, carriage of Salmonella enterica, and antimicrobial resistance genes in commercial pigs. 120 pigs on 2 commercial farms. A cohort of sixty 10-week-old pigs in 4 pens/farm (15 pigs/pen) was randomly selected. Equal numbers of pigs were given feed containing tylosin (40 μg/g of feed) for 0, 6, or 12 weeks. C-reactive protein concentrations were measured, microbial culture for S enterica in feces was performed, and antimicrobial resistance genes in feces were quantified. No significant associations were detected between C-reactive protein concentration or S enterica status and tylosin treatment. During the 12 weeks of tylosin administration, increased levels of 6 antimicrobial resistance genes did not occur. Treatment of pigs with tylosin did not affect C-reactive protein concentration or reduce carriage or load of S enterica. There was no evidence that pigs receiving tylosin had increased carriage of the 6 antimicrobial resistance genes measured. S enterica is a public health concern. Use of the antimicrobial growth promoter tylosin did not pose a public health risk by means of increased carriage of S enterica.
Sandeepa M Eswarappa
Full Text Available The genus Salmonella includes many pathogens of great medical and veterinary importance. Bacteria belonging to this genus are very closely related to those belonging to the genus Escherichia. lacZYA operon and lacI are present in Escherichia coli, but not in Salmonella enterica. It has been proposed that Salmonella has lost lacZYA operon and lacI during evolution. In this study, we have investigated the physiological and evolutionary significance of the absence of lacI in Salmonella enterica. Using murine model of typhoid fever, we show that the expression of LacI causes a remarkable reduction in the virulence of Salmonella enterica. LacI also suppresses the ability of Salmonella enterica to proliferate inside murine macrophages. Microarray analysis revealed that LacI interferes with the expression of virulence genes of Salmonella pathogenicity island 2. This effect was confirmed by RT-PCR and Western blot analysis. Interestingly, we found that SBG0326 of Salmonella bongori is homologous to lacI of Escherichia coli. Salmonella bongori is the only other species of the genus Salmonella and it lacks the virulence genes of Salmonella pathogenicity island 2. Overall, our results demonstrate that LacI is an antivirulence factor of Salmonella enterica and suggest that absence of lacI has facilitated the acquisition of virulence genes of Salmonella pathogenicity island 2 in Salmonella enterica making it a successful systemic pathogen.
José Pablo Soto-Arias
Full Text Available Recently, most foodborne illness outbreaks of salmonellosis have been caused by consumption of contaminated fresh produce. Yet, the mechanisms that allow the human pathogen Salmonella enterica to contaminate and grow in plant environments remain poorly described. We examined the effect of feeding by phytophagous insects on survival of S. enterica on lettuce. Larger S. enterica populations were found on leaves infested with Macrosteles quadrilineatus. In contrast, pathogen populations among plants exposed to Frankliniella occidentalis or Myzus persicae were similar to those without insects. However, on plants infested with F. occidentalis, areas of the infested leaf with feeding damage sustained higher S. enterica populations than areas without damage. The spatial distribution of S. enterica cells on leaves infested with F. occidentalis may be altered resulting in higher populations in feeding lesions or survival may be different across a leaf dependent on local damage. Results suggest the possibility of some specificity with select insects and the persistence of S. enterica. Additionally, we demonstrated the potential for phytophagous insects to become contaminated with S. enterica from contaminated plant material. S. enterica was detected in approximately 50% of all M. quadrilineatus, F. occidentalis, and M. persicae after 24 h exposure to contaminated leaves. Particularly, 17% of F. occidentalis, the smallest of the insects tested, harbored more than 10(2 CFU/F. occidentalis. Our results show that phytophagous insects may influence the population dynamics of S. enterica in agricultural crops. This study provides evidence of a human bacterial pathogen interacting with phytophagous insect during plant infestation.
Soto-Arias, José Pablo; Groves, Russell; Barak, Jeri D
Recently, most foodborne illness outbreaks of salmonellosis have been caused by consumption of contaminated fresh produce. Yet, the mechanisms that allow the human pathogen Salmonella enterica to contaminate and grow in plant environments remain poorly described. We examined the effect of feeding by phytophagous insects on survival of S. enterica on lettuce. Larger S. enterica populations were found on leaves infested with Macrosteles quadrilineatus. In contrast, pathogen populations among plants exposed to Frankliniella occidentalis or Myzus persicae were similar to those without insects. However, on plants infested with F. occidentalis, areas of the infested leaf with feeding damage sustained higher S. enterica populations than areas without damage. The spatial distribution of S. enterica cells on leaves infested with F. occidentalis may be altered resulting in higher populations in feeding lesions or survival may be different across a leaf dependent on local damage. Results suggest the possibility of some specificity with select insects and the persistence of S. enterica. Additionally, we demonstrated the potential for phytophagous insects to become contaminated with S. enterica from contaminated plant material. S. enterica was detected in approximately 50% of all M. quadrilineatus, F. occidentalis, and M. persicae after 24 h exposure to contaminated leaves. Particularly, 17% of F. occidentalis, the smallest of the insects tested, harbored more than 10(2) CFU/F. occidentalis. Our results show that phytophagous insects may influence the population dynamics of S. enterica in agricultural crops. This study provides evidence of a human bacterial pathogen interacting with phytophagous insect during plant infestation.
Michael B Cooley
Full Text Available 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 two 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 1,386 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 1,405 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 1,405 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 towards an understanding of transport leading to produce
Arredondo-García, José Luis; Calderón, Ernesto; Echániz-Aviles, Gabriela; Soto-Noguerón, Araceli; Arzate, Patricia; Amabile-Cuevas, Carlos F
Streptococcus pneumoniae is a worldwide leading cause of morbidity and mortality, while susceptibility towards penicillin and macrolides can be less than 50% in many regions. A total of 150 isolates of S. pneumoniae causative of invasive diseases in children were characterized, of which 24.6% had a fatal outcome. The most prevalent serotypes were 19F, 6B, 23F and 14. Resistance to penicillin, erythromycin (mostly of macrolide-lincosamide-streptogramin resistance phenotype) or trimethoprim-sulfamethoxazole was found in more than 40% of the isolates, but no resistance phenotype appeared linked to lethality. Serotype 3 isolates, which were seldom resistant, had a twofold lethality rate compared to the total sample. Serotyping could provide a better outcome-predicting tool than susceptibility testing. The seven-valent vaccine does not include the most prevalent serotypes found in Mexico.
Nayarit-Ballesteros, Nova; Rubio-Lozano, María Salud; Delgado-Suárez, Enrique; Méndez-Medina, Danilo; Braña-Varela, Diego; Rodas-Suárez, Oscar
To determine the serotype and antibiotic resistance profile of Salmonella spp. isolated from retail ground beef in Mexico City. A total of 100 samples of ground beef were analyzed. The pathogen was isolated by conventional methods and confirmed by PCR (invA gene, 284 bp).The antibiotic resistance profile was determined by the Kirby-Bauer method while serotyping was performed according to the Kauffman-White scheme. We isolated a total of 19 strains of Lomita (6), Derby (4), Senftenberg (2), Javiana and Cannsttat (1) and undetermined (5) serotypes. The strains showed a high resistance rate to ampicillin (18/19), carbenicillin (16/19), tetracyclin (13/19), and trimethoprim-sulfamethoxazole (13/19). Multidrug resistance was observed in 14 isolates. Several Salmonella spp. serotypes of public health significance are circulating in ground beef sold in the major Mexican city. Some of these strains are multi-drug resistance.
Heckenberg, S. G. B.; Brouwer, M. C.; van der Ende, A.; Hensen, E. F.; van de Beek, D.
Clin Microbiol Infect 2012; 18: 849855 Abstract We assessed the incidence of hearing loss and its relationship with clinical characteristics and pneumococcal serotypes in adults surviving pneumococcal meningitis. We analysed hearing loss in 531 adults surviving pneumococcal meningitis included in
Metzgar, David; Osuna, Miguel; Yingst, Samuel; Rakha, Magda; Earhart, Kenneth; Elyan, Diaa; Esmat, Hala; Saad, Magdi D; Kajon, Adriana; Wu, Jianguo; Gray, Gregory C; Ryan, Margaret A; Russell, Kevin L
.... Species and serotype identities were determined using several well-validated multiplex PCR protocols culled from the literature and supplemented with a few novel primer sets designed to identify rare types...
Harboe, Zitta B; Valentiner-Branth, Palle; Ingels, Helene
A seven-valent pneumococcal conjugate vaccine (PCV7) was introduced in the Danish childhood immunization program (2+1 schedule) in October 2007, followed by PCV13 starting from April 2010. The nationwide incidence of IPD among children younger than 5 years nearly halved after the introduction...... of children suspected to present with a vaccine failure. The period between April 19 and December 31, 2010 was considered a PCV7/PCV13 transitional period, where both vaccines were offered. We identified 45 episodes of IPD caused by a PCV7 serotype (23% of the total number) and 105 (55%) caused by one...... of the 6 additional serotypes in PCV13. Ten children had received at least one PCV7 dose before the onset of IPD caused by a PCV7 serotype. Seven children were considered to be incompletely vaccinated before IPD, but only three cases fulfilled the criteria of vaccine failure (caused by serotypes 14, 19F...
Full Text Available Objective. To determine the serotype and antibiotic resistance profile of Salmonella spp. isolated from retail ground beef in Mexico City. Materials and methods. A total of 100 samples of ground beef were analyzed. The pathogen was isolated by conventional methods and confirmed by PCR (invA gene, 284 bp. The antibiotic resistance profile was determined by the Kirby-Bauer method while serotyping was performed according to the Kauffman-White scheme. Results. We isolated a total of 19 strains of Lomita (6, Derby (4, Senftenberg (2, Javiana and Cannsttat (1 and undeter- mined (5 serotypes. The strains showed a high resistance rate to ampicillin (18/19, carbenicillin (16/19, tetracyclin (13/19, and trimethoprim-sulfamethoxazole (13/19. Multidrug resistance was observed in 14 isolates. Conclusions. Several Salmonella spp. serotypes of public health significance are circulating in ground beef sold in the major Mexican city. Some of these strains are multi-drug resistance.
Sharma-Chawla, Niharika; Sender, Vicky; Kershaw, Olivia; Gruber, Achim D.; Volckmar, Julia; Henriques-Normark, Birgitta
Influenza A virus (IAV) and Streptococcus pneumoniae are major causes of respiratory tract infections, particularly during coinfection. The synergism between these two pathogens is characterized by a complex network of dysregulated immune responses, some of which last until recovery following IAV infection. Despite the high serotype diversity of S. pneumoniae and the serotype replacement observed since the introduction of conjugate vaccines, little is known about pneumococcal strain dependency in the enhanced susceptibility to severe secondary S. pneumoniae infection following IAV infection. Thus, we studied how preinfection with IAV alters host susceptibility to different S. pneumoniae strains with various degrees of invasiveness using a highly invasive serotype 4 strain, an invasive serotype 7F strain, and a carrier serotype 19F strain. A murine model of pneumococcal coinfection during the acute phase of IAV infection showed a significantly increased degree of pneumonia and mortality for all tested pneumococcal strains at otherwise sublethal doses. The incidence and kinetics of systemic dissemination, however, remained bacterial strain dependent. Furthermore, we observed strain-specific alterations in the pulmonary levels of alveolar macrophages, neutrophils, and inflammatory mediators ultimately affecting immunopathology. During the recovery phase following IAV infection, bacterial growth in the lungs and systemic dissemination were enhanced in a strain-dependent manner. Altogether, this study shows that acute IAV infection predisposes the host to lethal S. pneumoniae infection irrespective of the pneumococcal serotype, while the long-lasting synergism between IAV and S. pneumoniae is bacterial strain dependent. These results hold implications for developing tailored therapeutic treatment regimens for dual infections during future IAV outbreaks. PMID:27647871
Nguyen, Bach Hoang; Phan, Dieu Hong Nu; Nguyen, Hien Xuan; Le, An Van; Alberti, Alberto
Streptococcus suis (S. suis) serotype 2 has recently become the most prevalent cause of meningitis in adults in many areas of Vietnam. This study provides data on S. suis molecular diagnosis in central Vietnam using a real-time polymerase chain reaction (PCR) assay targeting the S. suis serotype 2 cps2J gene. Additionally, 16S-23S rDNA intragenic spacer (ITS)-based phylogenic analysis of strains isolated from cerebrospinal fluid (CSF) in Thua Thien Hue Province, Vietnam, is presented and discussed. Pathogenic bacteria were isolated from 40 CSF samples, and 18 were identified as S. suis by culture-dependent methods. Capsular serotyping was assessed by real-time PCR. ITS sequences were obtained after traditional PCR and were used in phylogenic analyses. Pathogenic bacteria were isolated from 36 out of 40 CSF samples. A total of 18 S. suis strains were isolated and assigned to serotype 2 by real-time PCR. One CSF sample, negative when tested by culture-dependent methods, was positive to S. suis serotype 2 by real-time PCR. Pairwise alignments of the 18 ITS sequences did not reveal any variable nucleotide position, and resulted in a single sequence type. Sequences were similar to S. suis serotype 2 reference ITS sequences (> 98.1%), and there was no lack of an ITS spacer region in the isolates. S. suis serotype 2 is the most prevalent serotype in central Vietnam. Real-time PCR assay proved to be a reliable diagnostic method for early detection of S. suis 2 in CSF samples.
Harboe, Zitta B; Thomsen, Reimar W; Riis, Anders
younger than age 5 y. Age, male sex, meningitis, high comorbidity level, alcoholism, and early decade of diagnosis were significantly associated with mortality. Among individuals aged 5 y and older, serotypes 31, 11A, 35F, 17F, 3, 16F, 19F, 15B, and 10A were associated with highly increased mortality...... as compared with serotype 1 (all: adjusted odds ratio >or=3, p
Andresen, Lars Ole; Tegtmeier, Conny
The safety and protective efficacy of a horse antiserum raised against inactivated whole cell preparations of Streptococcus suis serotype 2 was investigated in pigs by experimental challenge. The antiserum was evaluated in two similar experiments each comprising 12 4-week-old pigs treated with 6 ...... indicate that passive immunization of pigs may be a way to reduce or control S. suis serotype 2 infections in pigs....
Pan, Zihao; Ma, Jiale; Dong, Wenyang; Song, Wenchao; Wang, Kaicheng; Lu, Chengping; Yao, Huochun
Streptococcus suis is an emerging zoonotic pathogen causing severe infections in pigs and humans. In previous studies, 33 serotypes of S. suis have been identified using serum agglutination. Here, we describe a novel S. suis strain, CZ130302, isolated from an outbreak of acute piglet meningitis in eastern China. Strong pathogenicity of meningitis caused by strain CZ130302 was reproduced in the BALB/c mouse model. The strain showed a high fatality rate (8/10), higher than those for known virulent serotype 2 strains P1/7 (1/10) and 9801 (2/10). Cell adhesion assay results with bEnd.3 and HEp2 cells showed that CZ130302 was significantly close to P1/7 and 9801. Both the agglutination test and its complementary test showed that strain CZ130302 had no strong cross-reaction with the other 33 S. suis serotypes. The multiplex PCR assays revealed no specified bands for all four sets used to detect the other 33 serotypes. In addition, genetic analysis of the whole cps gene clusters of all serotypes was performed in this study. The results of comparative genomics showed that the cps gene cluster of CZ130302, which was not previously reported, showed no homology to the gene sequences of the other strains. Especially, the wzy, wzx, and acetyltransferase genes of strain CZ130302 are phylogenetically distinct from strains of the other 33 serotypes. Therefore, this study suggested that strain CZ130302 represents a novel variant serotype of S. suis (designated serotype Chz) which has a high potential to be virulent and associated with meningitis in animals. Copyright © 2015, American Society for Microbiology. All Rights Reserved.
Li, Xiaoxin; Bangari, Dinesh S.; Sharma, Anurag; Mittal, Suresh K.
Bovine adenovirus serotype 3 (BAd3) and porcine adenovirus serotype 3 (PAd3) entry into the host cells is independent of Coxsackievirus -adenovirus receptor and integrins. The role of sialic acid in BAd3 and PAd3 entry was investigated. Removal of sialic acid by neuraminidase, or blocking sialic acid by wheat germ agglutinin lectin significantly inhibited BAd3, but not PAd3, transduction of Madin Darby bovine kidney cells. Maackia amurensis agglutinin or Sambucus nigra (elder) agglutinin trea...
Full Text Available Vibrio parahaemolyticus has emerged as a major foodborne pathogen in China, Japan, Thailand and other Asian countries. In this study, 72 strains of V. parahaemolyticus were isolated from clinical and environmental samples between 2006 and 2014 in Jiangsu, China. The serotypes and six virulence genes including thermostable direct hemolysin (TDR and TDR-related hemolysin (TRH genes were assessed among the isolates. Twenty five serotypes were identified and O3:K6 was one of the dominant serotypes. The genetic diversity was assessed by multilocus sequence typing (MLST analysis, and 48 sequence types (STs were found, suggesting this V. parahaemolyticus group is widely dispersed and undergoing rapid evolution. A total of 25 strains of pandemic serotypes such as O3:K6, O5:K17 and O1:KUT were identified. It is worth noting that the pandemic serotypes were not exclusively identified from clinical samples, rather, nine strains were also isolated from environmental samples; and some of these strains harbored several virulence genes, which may render those strains pathogenicity potential. Therefore, the emergence of these environmental pandemic V. parahaemolyticus strains may poses a new threat to the public health in China. Furthermore, six novel serotypes and 34 novel STs were identified among the 72 isolates, indicating that V. parahaemolyticus were widely distributed and fast evolving in the environment in Jiangsu, China. The findings of this study provide new insight into the phylogenic relationship between V. parahaemolyticus strains of pandemic serotypes from clinical and environmental sources and enhance the MLST database; and our proposed possible O- and K- antigen evolving paths of V. parahaemolyticus may help understand how the serotypes of this dispersed bacterial population evolve.
Nova Nayarit-Ballesteros; María Salud Rubio-Lozano; Enrique Delgado-Suárez; Danilo Méndez-Medina; Diego Braña-Varela; Oscar Rodas-Suárez
Objective. To determine the serotype and antibiotic resistance profile of Salmonella spp. isolated from retail ground beef in Mexico City. Materials and methods. A total of 100 samples of ground beef were analyzed. The pathogen was isolated by conventional methods and confirmed by PCR (invA gene, 284 bp). The antibiotic resistance profile was determined by the Kirby-Bauer method while serotyping was performed according to the Kauffman-White scheme. Results. We isolated a total of 19 strains o...
Fussing, V.; Møller Nielsen, E.; Neimann, J.
In order to monitor the distribution of subtypes of Campylobacter and to identify clusters, 975 isolates of Campylobacter spp., obtained from human infections occurring in two Danish counties, were studied during a 1-year period. The isolates were characterised by Penner serotyping and automated ...... the validity of selected clusters identified by serotyping and ribotyping. The observed clustering of Campylobacter isolates, with identical types in time and place, indicates that common-source outbreaks of campylobacteriosis are more common than is usually thought....
Valarcher, Jean-Francois; Knowles, Nick J.; Zakharov, Valery; Scherbakov, Alexey; Zhang, Zhidong; Shang, You-Jun; Liu, Zai-Xin; Liu, Xiang-Tao; Sanyal, Aniket; Hemadri, Divakar; Tosh, Chakradhar; Rasool, Thaha J.; Pattnaik, Bramhadev; Schumann, Kate R.; Beckham, Tammy R.
We investigated the molecular epidemiology of foot-and-mouth disease virus (FMDV) serotype Asia 1, which caused outbreaks of disease in Asia during 2003?2007. Since 2004, the region affected by outbreaks of this serotype has increased from disease-endemic countries in southern Asia (Afghanistan, India, Iran, Nepal, Pakistan) northward to encompass Kyrgyzstan, Tajikistan, Uzbekistan, several regions of the People?s Republic of China, Mongolia, Eastern Russia, and North Korea. Phylogenetic anal...
Halsey, Eric S.; Marks, Morgan A.; Gotuzzo, Eduardo; Fiestas, Victor; Suarez, Luis; Vargas, Jorge; Aguayo, Nicolas; Madrid, Cesar; Vimos, Carlos; Kochel, Tadeusz J.; Laguna-Torres, V. Alberto
Background Disease caused by the dengue virus (DENV) is a significant cause of morbidity throughout the world. Although prior research has focused on the association of specific DENV serotypes (DENV-1, DENV-2, DENV-3, and DENV-4) with the development of severe outcomes such as dengue hemorrhagic fever and dengue shock syndrome, relatively little work has correlated other clinical manifestations with a particular DENV serotype. The goal of this study was to estimate and compare the prevalence of non-hemorrhagic clinical manifestations of DENV infection by serotype. Methodology and Principal Findings Between the years 2005–2010, individuals with febrile disease from Peru, Bolivia, Ecuador, and Paraguay were enrolled in an outpatient passive surveillance study. Detailed information regarding clinical signs and symptoms, as well as demographic information, was collected. DENV infection was confirmed in patient sera with polyclonal antibodies in a culture-based immunofluorescence assay, and the infecting serotype was determined by serotype-specific monoclonal antibodies. Differences in the prevalence of individual and organ-system manifestations were compared across DENV serotypes. One thousand seven hundred and sixteen individuals were identified as being infected with DENV-1 (39.8%), DENV-2 (4.3%), DENV-3 (41.5%), or DENV-4 (14.4%). When all four DENV serotypes were compared with each other, individuals infected with DENV-3 had a higher prevalence of musculoskeletal and gastrointestinal manifestations, and individuals infected with DENV-4 had a higher prevalence of respiratory and cutaneous manifestations. Conclusions/Significance Specific clinical manifestations, as well as groups of clinical manifestations, are often overrepresented by an individual DENV serotype. PMID:22563516
Handberg, Kurt; Nielsen, Ole L.; Jørgensen, Poul Henrik
was applied to samples collected from four commercial table egg layer flocks of young stock or pullets vaccinated with either serotype 1 (CVI988) or serotype 3 (HVT) vaccine. These flocks had various clinical signs of Marek's disease. MDV-1 was detected in buffy-coat cells, spleen, liver, skin, feather tips...... and ovaries. The detection of MDV in feather tips appeared to be as sensitive as co-cultivation of buffy-coat cells, although an inhibiting factor was observed in extracts from feather tips of non-white chickens. This inhibition could be overcome in most extracts by applying a bovine serum albumen...
Full Text Available Shigella flexneri is the major pathogen causing bacillary dysentery in developing countries. S. flexneri is divided into at least 16 serotypes based on the combination of antigenic determinants present in the O-antigen. All the serotypes (except for serotype 6 share a basic O-unit containing one N-acetyl-d-glucosamine and three l-rhamnose residues, whereas differences between the serotypes are conferred by phage-encoded glucosylation and/or O-acetylation. Serotype Xv is a newly emerged and the most prevalent serotype in China, which can agglutinate with both MASF IV-1 and 7,8 monoclonal antibodies. The factor responsible for the presence of MASF IV-1 (E1037 epitope has not yet been identified. In this study, we analyzed the LPS structure of serotype Xv strains and found that the MASF IV-1 positive phenotype depends on an O-antigen modification with a phosphoethanolamine (PEtN group attached at position 3 of one of the rhamnose residues. A plasmid carried gene, lpt-O (LPS phosphoethanolamine transferase for O-antigen, mediates the addition of PEtN for serotype Xv and other MASF IV-1 positive strains. These findings reveal a novel serotype conversion mechanism in S. flexneri and show the necessity of further extension of the serotype classification scheme recognizing the MASF IV-1 positive strains as distinctive subtypes.
Uddin, Reaz; Sufian, Muhammad
Infections caused by Salmonella enterica, a Gram-negative facultative anaerobic bacteria belonging to the family of Enterobacteriaceae, are major threats to the health of humans and animals. The recent availability of complete genome data of pathogenic strains of the S. enterica gives new avenues for the identification of drug targets and drug candidates. We have used the genomic and metabolic pathway data to identify pathways and proteins essential to the pathogen and absent from the host. We took the whole proteome sequence data of 42 strains of S. enterica and Homo sapiens along with KEGG-annotated metabolic pathway data, clustered proteins sequences using CD-HIT, identified essential genes using DEG database and discarded S. enterica homologs of human proteins in unique metabolic pathways (UMPs) and characterized hypothetical proteins with SVM-prot and InterProScan. Through this core proteomic analysis we have identified enzymes essential to the pathogen. The identification of 73 enzymes common in 42 strains of S. enterica is the real strength of the current study. We proposed all 73 unexplored enzymes as potential drug targets against the infections caused by the S. enterica. The study is comprehensive around S. enterica and simultaneously considered every possible pathogenic strain of S. enterica. This comprehensiveness turned the current study significant since, to the best of our knowledge it is the first subtractive core proteomic analysis of the unique metabolic pathways applied to any pathogen for the identification of drug targets. We applied extensive computational methods to shortlist few potential drug targets considering the druggability criteria e.g. Non-homologous to the human host, essential to the pathogen and playing significant role in essential metabolic pathways of the pathogen (i.e. S. enterica). In the current study, the subtractive proteomics through a novel approach was applied i.e. by considering only proteins of the unique metabolic
Marluísa de Oliveira Guimarães Ishak
Full Text Available INTRODUCTION: Chlamydia infection is associated with debilitating human diseases including trachoma, pneumonia, coronary heart disease and urogenital diseases. Serotypes of C. trachomatis show a fair correlation with the group of diseases they cause, and their distribution follows a well-described geographic pattern. Serotype A, a trachoma-associated strain, is known for its limited dissemination in the Middle East and Northern Africa. However, knowledge on the spread of bacteria from the genus Chlamydia as well as the distribution of serotypes in Brazil is quite limited. METHODS: Blood samples of 1,710 individuals from ten human population groups in the Amazon region of Brazil were examined for antibodies to Chlamydia using indirect immunofluorescence and microimmunofluorescence assays. RESULTS: The prevalence of antibodies to Chlamydia ranged from 23.9% (Wayana-Apalai to 90.7% (Awa-Guaja with a mean prevalence of 50.2%. Seroreactivity was detected to C. pneumoniae and to all serotypes of C. trachomatis tested; furthermore, we report clear evidence of the as-yet-undescribed occurrence of serotype A of C. trachomatis. CONCLUSIONS: Specific seroreactivity not only accounts for the large extent of dissemination of C. trachomatis in the Amazon region of Brazil but also shows an expanded area of occurrence of serotype A outside the epidemiological settings previously described. Furthermore, these data suggest possible routes of Chlamydia introduction into the Amazon region from the massive human migration that occurred during the 1,700s.
Full Text Available ABSTRACT The aim of this work was to study the prevalence of Listeria monocytogenes in foods obtained in retail shops and food industries located in Montevideo-Uruguay, and to identify the serogroups of the obtained isolates. Three-thousand one-hundred and seventy-five food samples (frozen, deli meats, ready-to-eat and cheese were analyzed. The obtained isolates were serogrouped by multiplex PCR and serotyped by conventional procedure. Genetic comparisons were performed using pulsed-field gel electrophoresis on a sub-set of isolates belonging to the same serotype successively recovered from the same establishment. L. monocytogenes was isolated from 11.2% of samples. The highest prevalence was observed in frozen foods (38%, followed by cheese (10%. 1/2b and 4b were the most frequently identified serotypes. In six of 236 analyzed establishments we successively recovered L. monocytogenes isolates belonging to the same serotype. Most of them corresponded to serotype 1/2b. Pulsed-field gel electrophoresis profiles suggest that at least 33% of L. monocytogenes 1/2b isolates are genetically related and that may remain viable for prolonged periods. The observed prevalence of L. monocytogenes was lower than reported in neighboring countries. Our findings highlight the role that frozen foods may play in the spread of this pathogen, and the relevance of serotypes 1/2b and 4b.
Stockmann, Chris; Ampofo, Krow; Pavia, Andrew T; Blaschke, Anne J; Mason, Edward O; Presson, Angela P; Forney, Larry J; Byington, Carrie L
The Red Queen hypothesis is an evolutionary theory that describes the reciprocal coevolution of competing species. We sought to study whether introduction of the 7- and 13-valent pneumococcal conjugate vaccines (PCV7 and PCV13, respectively) altered pneumococcal serotype dynamics among children with invasive pneumococcal disease (IPD) as predicted by the Red Queen hypothesis. This study examined pneumococcal isolates (n = 641) obtained from children <18 years of age hospitalized with IPD from 1997 to 2014 in Utah. A review of the literature also identified several additional studies conducted in the United States and Europe that were used to test the external generalizability of our Utah findings. Simpson's index was used to quantify pneumococcal serotype diversity. In Utah, the introduction of PCV7 and PCV13 was associated with rapid increases in serotype diversity (P < .001). Serotypes rarely present before vaccine introduction emerged as common causes of IPD. Diversity then decreased (P < .001) as competition selected for the fittest serotypes and new evolutionary equilibriums were established. This pattern was also observed more broadly in the United States, the United Kingdom, Norway, and Spain. This vaccine-driven example of human/bacterial coevolution appears to confirm the Red Queen hypothesis, which reveals a limitation of serotype-specific vaccines and offers insights that may facilitate alternative strategies for the elimination of IPD. © The Author 2016. Published by Oxford University Press for the Infectious Diseases Society of America. All rights reserved. For permissions, e-mail email@example.com.
Ukae, S; Nakata, S; Adachi, N; Kogawa, K; Chiba, S
We conducted a clinical trial of rhesus rotavirus vaccine MMU-18006 (RRV, serotype 3) to assess the immunogenicity, transmissibility and booster effect of this vaccine in a welfare nursery in Sapporo, from September 1986 to October 1988. After the trial, in March 1989, an outbreak of gastroenteritis due to a wild strain of serotype 1 rotavirus (RV-1) occurred in the study population. Infants were divided into three groups based on vaccination history: five booster vaccinees, 18 one-dose vaccinees and 18 control infants who did not receive vaccine. There was a significant relationship between asymptomatic infection and higher levels of preoutbreak antibody titres against KU (serotype 1) but not RRV. Significant protection from rotavirus illness was observed both in the booster vaccine group and in the one-dose vaccine group but not in the control group. Rotavirus-specific serum IgA immune response was considered to be one of the indicators of recent rotavirus infection, and did not correlate with resistance to rotavirus illness. Our results revealed that protection from rotavirus illness was serotype-specific and that previous rotavirus infection, including vaccination, was important to induce the heterotypic immune response, and that ageing or booster inoculation of RRV might play a role in the protection against serotype 1 rotavirus infection. From our findings, a booster administration was thought to be important to induce effective heterotypic immunity and should be included in a future rotavirus vaccine trial to obtain sufficient protection against four major serotypes of rotavirus.
Cheaib, Miriam; Dehghani Amirabad, Azim; Nordström, Karl J V; Schulz, Marcel H; Simon, Martin
Phenotypic variation of a single genotype is achieved by alterations in gene expression patterns. Regulation of such alterations depends on their time scale, where short-time adaptations differ from permanently established gene expression patterns maintained by epigenetic mechanisms. In the ciliate Paramecium, serotypes were described for an epigenetically controlled gene expression pattern of an individual multigene family. Paradoxically, individual serotypes can be triggered in Paramecium by alternating environments but are then stabilized by epigenetic mechanisms, thus raising the question to which extend their expression follows environmental stimuli. To characterize environmental adaptation in the context of epigenetically controlled serotype expression, we used RNA-seq to characterize transcriptomes of serotype pure cultures. The resulting vegetative transcriptome resource is first analysed for genes involved in the adaptive response to the altered environment. Secondly, we identified groups of genes that do not follow the adaptive response but show co-regulation with the epigenetically controlled serotype system, suggesting that their gene expression pattern becomes manifested by similar mechanisms. In our experimental set-up, serotype expression and the entire group of co-regulated genes were stable among environmental changes and only heat-shock genes altered expression of these gene groups. The data suggest that the maintenance of these gene expression patterns in a lineage represents epigenetically controlled robustness counteracting short-time adaptation processes. © The Author 2015. Published by Oxford University Press on behalf of Kazusa DNA Research Institute.
Guo, Xi; Wang, Min; Wang, Lu; Wang, Yao; Chen, Tingting; Wu, Pan; Chen, Min; Liu, Bin; Feng, Lu
Serotyping based on surface polysaccharide antigens is important for the clinical detection and epidemiological surveillance of pathogens. Polysaccharide gene clusters (PSgcs) are typically responsible for the diversity of bacterial surface polysaccharides. Through whole-genome sequencing and analysis, eight putative PSgc types were identified in 23 Enterobacter aerogenes strains from several geographic areas, allowing us to present the first molecular serotyping system for E. aerogenes . A conventional antigenic scheme was also established and correlated well with the molecular serotyping system that was based on PSgc genetic variation, indicating that PSgc-based molecular typing and immunological serology provide equally valid results. Further, a multiplex Luminex-based array was developed, and a double-blind test was conducted with 97 clinical specimens from Shanghai, China, to validate our array. The results of these analyses indicated that strains containing PSgc4 and PSgc7 comprised the predominant groups. We then examined 86 publicly available E. aerogenes strain genomes and identified an additional seven novel PSgc types, with PSgc10 being the most abundant type. In total, our study identified 15 PSgc types in E. aerogenes , providing the basis for a molecular serotyping scheme. From these results, differing epidemic patterns were identified between strains that were predominant in different regions. Our study highlights the feasibility and reliability of a serotyping system based on PSgc diversity, and for the first time, presents a molecular serotyping system, as well as an antigenic scheme for E. aerogenes , providing the basis for molecular diagnostics and epidemiological surveillance of this important emerging pathogen.
Llanes, Rafael; Lazo, Alcides; Somarriba, Lorenzo; Mas, Pedro
Over 700,000 cases of cholera were reported in Haiti between October 2010 and February 2015. In November 2011, the Cuban Medical Team serving in Haiti established a laboratory-supported sentinel surveillance system for cholera in 10 public hospitals (one in each of Haiti's 10 departments), to estimate the proportion of hospitalized patients with cholera and detect emergence of new Vibrio cholerae serotypes. Each month, the first ten stool samples collected from patients admitted with acute watery diarrhea were studied in all hospitals involved. Surveillance system findings from November 1, 2011, to October 30, 2012 showed that acute watery diarrhea was caused by V. cholerae serogroup O1 in 45.9% (210/458) of patients: Serotype Ogawa was found in 98.6% of this isolates (207/210) and serotype Inaba in 1.4% (3/210), indicating low circulation level of the latter in Haiti. Continuing laboratory sentinel surveillance of V. cholerae is needed to monitor the spread of the disease and prevent and contain outbreaks, particularly of new serotypes. It is important to ensure that these findings are systematically integrated with data available to MSPP from other surveillance sources. KEYWORDS Vibrio cholerae, serotype Inaba, serotype Ogawa, epidemiological surveillance, medical cooperation, Haiti, Cuba.
Gentsch, Jon R; Laird, Ashley R; Bielfelt, Brittany; Griffin, Dixie D; Banyai, Krisztian; Ramachandran, Madhu; Jain, Vivek; Cunliffe, Nigel A; Nakagomi, Osamu; Kirkwood, Carl D; Fischer, Thea K; Parashar, Umesh D; Bresee, Joseph S; Jiang, Baoming; Glass, Roger I
The development of rotavirus vaccines that are based on heterotypic or serotype-specific immunity has prompted many countries to establish programs to assess the disease burden associated with rotavirus infection and the distribution of rotavirus strains. Strain surveillance helps to determine whether the most prevalent local strains are likely to be covered by the serotype antigens found in current vaccines. After introduction of a vaccine, this surveillance could detect which strains might not be covered by the vaccine. Almost 2 decades ago, studies demonstrated that 4 globally common rotavirus serotypes (G1-G4) represent >90% of the rotavirus strains in circulation. Subsequently, these 4 serotypes were used in the development of reassortant vaccines predicated on serotype-specific immunity. More recently, the application of reverse-transcription polymerase chain reaction genotyping, nucleotide sequencing, and antigenic characterization methods has confirmed the importance of the 4 globally common types, but a much greater strain diversity has also been identified (we now recognize strains with at least 42 P-G combinations). These studies also identified globally (G9) or regionally (G5, G8, and P2A) common serotype antigens not covered by the reassortant vaccines that have undergone efficacy trials. The enormous diversity and capacity of human rotaviruses for change suggest that rotavirus vaccines must provide good heterotypic protection to be optimally effective.
Abreu, Cândida; Silva-Pinto, André; Lazzara, Daniela; Sobrinho-Simões, Joana; Guimarães, João Tiago; Sarmento, António
All the reports from Angola's 2013 dengue outbreak revealed serotype 1. However, previously dengue serotypes 1-4 have been reported in Africa and in 2014 serotype 4 was reported in Angola. To report dengue serotypes in patients returning from Angola during 2013 outbreak. Retrospective, cross-sectional study. We serotyped the dengue by an in house Polymerase Chain Reaction technique in randomly selected cases. From the 2013 Angola's dengue outbreak we treated 47 adult patients. None had history of past dengue. A combo kit test for dengue revealed positive NS1 antigen in 39 and IgM antibodies in 8. From 17 randomly patients tested by RNA Real Time-PCR, 11 were positive: 7 for DENV-1, 2 for DENV-2, 1 for DENV-3 (co-infected with DENV-1) and 1 for DENV-4. None had a complicated or fatal evolution. Unlike previous reports the 4 serotypes were detected, and this resulted in a different epidemiological situation, raising the risk of future outbreaks of severe dengue. Copyright © 2016 Elsevier B.V. All rights reserved.
Harapas, Dean; Premier, Robert; Tomkins, Bruce; Hepworth, Graham; Ajlouni, Said
Minor shoot injury significantly (P cos lettuce in the greenhouse. Initial mean counts of the Salmonella on the injured and uninjured cos lettuce were on the order of 6 log CFU/g. After 3 days, the mean count decreased to 4.8 log CFU/g on the injured plants compared with the significantly (P cos lettuce and S. enterica serovar Sofia on chive. The findings reaffirm earlier results with Escherichia coli and increase the impetus to avoid shoot injury during the production of cos lettuce and chive, if bacteria of food safety concern are present.
Jensen, Annette Nygaard; Dalsgaard, Anders; Stockmarr, Anders
, which caused substantial contamination of the environment. This suggests that isolation of animals as soon as a Salmonella infection is indicated by clinical symptoms of diarrhea could be a means of reducing and controlling the spread and persistence of Salmonella in outdoor organic pig production......It was investigated how organic rearing conditions influence the Salmonella enterica infection dynamics in pigs and whether Salmonella persists in the paddock environment. Pigs inoculated with S. enterica serovar Typhimurium were grouped with Salmonella-negative tracer pigs. Bacteriological...... of Salmonella-negative pigs into four naturally Salmonella-contaminated paddocks caused Salmonella infections of pigs in two paddocks. In one of these paddocks, all tracer pigs (n = 10) became infected, coinciding with a previous high Salmonella infection rate and high Salmonella excretion level. Our results...
Full Text Available Abstract A total of 518 fecal samples collected from 183 apparently healthy cattle, 180 pigs and 155 broilers throughout Japan in 1999 were examined to determine the prevalence and antimicrobial susceptibility of Salmonella. The isolation rates were 36.1% in broilers, 2.8% in pigs and 0.5% in cattle. S. enterica Infantis was the most frequent isolate, found in 22.6% of broiler fecal samples. Higher resistance rates were observed against oxytetracycline (82.0%, dihydrostreptomycin (77.9%, kanamycin (41.0% and trimethoprim (35.2%. Resistance rates to ampicillin, ceftiofur, bicozamycin, chloramphenicol and nalidixic acid were S. enterica Senftenberg was found in the isolates obtained from one broiler fecal sample. This is the first report of cephalosporin-resistant Salmonella directly isolated from food animal in Japan.
Shachar, Dina; Yaron, Sima
Recent large foodborne outbreaks caused by Salmonella enterica serovars have been associated with consumption of foods with high fat content and reduced water activity, even though their ingredients usually undergo pasteurization. The present study was focused on the heat tolerance of Salmonella enterica serovars Agona, Enteritidis, and Typhimurium in peanut butter. The Salmonella serovars in the peanut butter were resistant to heat, and even at a temperature as high as 90 degrees C only 3.2-log reduction in CFU was observed. The obtained thermal inactivation curves were upwardly concave, indicating rapid death at the beginning (10 min) followed by lower death rates and an asymptotic tail. The curves fitted the nonlinear Weibull model with beta parameters pasteurization process cannot be improved significantly by longer treatment or higher temperatures.
Full Text Available Salmonella enterica serovar Typhi (S. Typhi causes typhoid fever, a disseminated infection, while the closely related pathogen S. enterica serovar Typhimurium (S. Typhimurium is associated with a localized gastroenteritis in humans. Here we investigated whether both pathogens differ in the chemotactic response they induce in neutrophils using a single-cell experimental approach. Surprisingly, neutrophils extended chemotactic pseudopodia toward Escherichia coli and S. Typhimurium, but not toward S. Typhi. Bacterial-guided chemotaxis was dependent on the presence of complement component 5a (C5a and C5a receptor (C5aR. Deletion of S. Typhi capsule biosynthesis genes markedly enhanced the chemotactic response of neutrophils in vitro. Furthermore, deletion of capsule biosynthesis genes heightened the association of S. Typhi with neutrophils in vivo through a C5aR-dependent mechanism. Collectively, these data suggest that expression of the virulence-associated (Vi capsular polysaccharide of S. Typhi obstructs bacterial-guided neutrophil chemotaxis.
Rounds, Joshua M; Hedberg, Craig W; Meyer, Stephanie; Boxrud, David J; Smith, Kirk E
We determined characteristics of Salmonella enterica pulsed-field gel electrophoresis clusters that predict their being solved (i.e., that result in identification of a confirmed outbreak). Clusters were investigated by the Minnesota Department of Health by using a dynamic iterative model. During 2001-2007, a total of 43 (12.5%) of 344 clusters were solved. Clusters of ≥4 isolates were more likely to be solved than clusters of 2 isolates. Clusters in which the first 3 case isolates were received at the Minnesota Department of Health within 7 days were more likely to be solved than were clusters in which the first 3 case isolates were received over a period >14 days. If resources do not permit investigation of all S. enterica pulsed-field gel electrophoresis clusters, investigation of clusters of ≥4 cases and clusters in which the first 3 case isolates were received at a public health laboratory within 7 days may improve outbreak investigations.
Löfström, Charlotta; Hintzmann, Ann-Sofie; Sørensen, Gitte
Salmonella enterica subspecies enterica serovar Typhimurium (S. Typhimurium) is one of the most prevalent serovars in Europe - where both poultry and poultry related products are common sources of human salmonellosis. Due to efficient control programs, the prevalence of S. Typhimurium in Danish...... poultry production is very low. Despite this, during the past decades there has been a reoccurring problem with infections with S. Typhimurium phage type DT41 in the Danish poultry production without identifying a clear source. In the end of 2013 and beginning of 2014 an increased isolation of S....... Typhimurium DT41 was noted mainly in this production, but also in other samples. To investigate this is in more detail, 47 isolates from egg layers (n = 5, 1 flock), broilers (n = 33, 13 flocks), broiler breeding flocks and hatches (n = 5; 2 flocks and 1 environmental hatchery sample), feed (n = 1), poultry...
Kwan, Grace; Charkowski, Amy O; Barak, Jeri D
Although enteric human pathogens are usually studied in the context of their animal hosts, a significant portion of their life cycle occurs on plants. Plant disease alters the phyllosphere, leading to enhanced growth of human pathogens; however, the impact of human pathogens on phytopathogen biology and plant health is largely unknown. To characterize the interaction between human pathogens and phytobacterial pathogens in the phyllosphere, we examined the interactions between Pectobacterium carotovorum subsp. carotovorum and Salmonella enterica or Escherichia coli O157:H7 with regard to bacterial populations, soft rot progression, and changes in local pH. The presence of P. carotovorum subsp. carotovorum enhanced the growth of both S. enterica and E. coli O157:H7 on leaves. However, in a microaerophilic environment, S. enterica reduced P. carotovorum subsp. carotovorum populations and soft rot progression by moderating local environmental pH. Reduced soft rot was not due to S. enterica proteolytic activity. Limitations on P. carotovorum subsp. carotovorum growth, disease progression, and pH elevation were not observed on leaves coinoculated with E. coli O157:H7 or when leaves were coinoculated with S. enterica in an aerobic environment. S. enterica also severely undermined the relationship between the phytobacterial population and disease progression of a P. carotovorum subsp. carotovorum budB mutant defective in the 2,3-butanediol pathway for acid neutralization. Our results show that S. enterica and E. coli O157:H7 interact differently with the enteric phytobacterial pathogen P. carotovorum subsp. carotovorum. S. enterica inhibition of soft rot progression may conceal a rapidly growing human pathogen population. Whereas soft rotted produce can alert consumers to the possibility of food-borne pathogens, healthy-looking produce may entice consumption of contaminated vegetables. Salmonella enterica and Escherichia coli O157:H7 may use plants to move between animal
Khan, Zaman; Rehman, Abdul; Hussain, Syed Z; Nisar, Muhammad A; Zulfiqar, Soumble; Shakoori, Abdul R
Cadmium resistant bacterium, isolated from industrial wastewater, was characterized as Salmonella enterica 43C on the basis of biochemical and 16S rRNA ribotyping. It is first ever reported S. enterica 43C bared extreme resistance against heavy metal consortia in order of Pb(2+)>Cd(2+)>As(3+)>Zn(2+)>Cr(6+)>Cu(2+)>Hg(2+). Cd(2+) stress altered growth pattern of the bacterium in time dependent manner. It could remove nearly 57 % Cd(2+) from the medium over a period of 8 days. Kinetic and thermodynamic studies based on various adsorption isotherm models (Langmuir and Freundlich) depicted the Cd(2+) biosorption as spontaneous, feasible and endothermic in nature. Interestingly, the bacterium followed pseudo first order kinetics, making it a good biosorbent for heavy metal ions. The S. enterica 43C Cd(2+) processivity was significantly influenced by temperature, pH, initial Cd(2+) concentration, biomass dosage and co-metal ions. FTIR analysis of the bacterium revealed the active participation of amide and carbonyl moieties in Cd(2+) adsorption confirmed by EDX analysis. Electron micrographs beckoned further surface adsorption and increased bacterial size due to intracellular Cd(2+) accumulation. An overwhelming increase in glutathione and other non-protein thiols levels played a significant role in thriving oxidative stress generated by metal cations. Presence of metallothionein clearly depicted the role of such proteins in bacterial metal resistance mechanism. The present study results clearly declare S. enterica 43C a suitable candidate for green chemistry to bioremediate environmental Cd(2+).
Full Text Available Aims: Salmonella enterica serovar Typhi is the major causative agent for typhoidial fever around the globe among human population reported till date. Present research work was carried out for detection and molecular characterisation of Salmonella enterica serovar Typhi isolated from humans with Typhoidial fever by biochemical, phenotypical and virulence gene based polymerase chain reaction (PCR techniques. The isolated strains were also investigated for antibiotic susceptibility patterns as a control measure. Methodology and Results: A total of 16 clinical samples were collected from the same numbers of patients (7 males and 9 females from Coimbatore, Erode and Salem districts of Tamil Nadu and were processed via broth enrichment methods for isolation and identification of the causative agent S. enterica serovar Typhi. Microbiological and biochemical investigations revealed the presence of S. Typhi from 16 samples. The biotyping of the isolates showed that all the isolates belonged to biotype IV. The PCR analysis confirmed the presence of invA (Invasion gene, 244bp, tyv (Tyveloseepimerase gene, 615 bp, fliC-d (Phage-1 flagellin gene for d-antigen, 750 bp and viaB (Vi antigen gene, 439bp in all 16 clinical samples. The antibiotic susceptibility test that was carried out among the isolates against 12 antimicrobial agents, showed 100 % resistance to only ampicillin and 100 % sensitivity to carbenicillin, chloramphenicol, clindamycin, gentamycin, kanamycin and tetracycline.Conclusion, significance and impact of study: This study confirmed the association of virulent strains of S. enterica serovar Typhi from Typhoidial fever among human population and suggested that PCR based diagnostic could be very useful for the rapid detection of S. Typhi isolates. Present study emphasized the use of antibiotic like chloramphenicol or in combination with other antibiotics for the effective control of S. Typhi.
Liakopoulos, Apostolos; Geurts, Yvon; Dierikx, Cindy M.; Brouwer, Mike; Kant, Arie; Wit, Ben; Heymans, Raymond; Pelt, Van Wilfrid; Mevius, Dik J.
Extended-spectrum cephalosporin-resistant Salmonella enterica serovar Heidelberg strains (JF6X01.0022/XbaI.0251, JF6X01.0326/XbaI.1966, JF6X01.0258/XbaI.1968, and JF6X01.0045/XbaI.1970) have been identified in the United States with pulsed-field gel electrophoresis. Our examination of isolates
Cloeckaert, Axel; Schwarz, Stefan
International audience; Multidrug-resistant Salmonella enterica serovar Typhimurium phage type DT104 has emerged during the last decade as a global health problem because of its involvement in diseases in animals and humans. Multidrug-resistant DT104 strains are mostly resistant to ampicillin, chloramphenicol, streptomycin, sulfonamides and tetracyclines (ACSSuT resistance type). The genes coding for such resistances are clustered on the chromosome. This paper reviews new developments in the ...
Zhou, Zhemin; Lundstrøm, Inge; Dien, Alicia Tran; Duchêne, Sebastián; Alikhan, Nabil-Fareed; Sergeant, Martin J.; Langridge, Gemma; Fotakis, Anna K.; Nair, Satheesh; Stenøien, Hans K.; Hamre, Stian S.; Casjens, Sherwood; Green, Monica H.; Christophersen, Axel; Quince, Christopher
Salmonella enterica serovar Paratyphi C causes enteric (paratyphoid) fever in humans. Its presentation can range from asymptomatic infections of the blood stream to gastrointestinal or urinary tract infection or even a fatal septicemia. Paratyphi C is very rare in Europe and North America, except for occasional travelers from South and East Asia or Africa where the disease is more common. However, early 20th century observations in Eastern Europe suggest that Paratyphi C enteric fever may onc...
Cameron-Veas, Karla; Fraile, Lorenzo; Napp, Sebastian; Garrido, Victoria; Grilló, María Jesús; Migura-Garcia, Lourdes
A longitudinal study was conducted to investigate the presence of multidrug antimicrobial resistance (multi-AR) in Salmonella enterica in pigs reared under conventional preventative medicine programmes in Spain and the possible association of multi-AR with ceftiofur or tulathromycin treatment during the pre-weaning period. Groups of 7-day-old piglets were treated by intramuscular injection with ceftiofur on four farms (n=40 piglets per farm) and with tulathromycin on another four farms (n=40 piglets per farm). A control group of untreated piglets (n=30 per farm) was present on each farm. Faecal swabs were collected for S. enterica culture prior to treatment, at 2, 7 and 180days post-treatment, and at slaughter. Minimal inhibitory concentrations of 14 antimicrobial agents, pulsed-field gel electrophoresis and detection of resistance genes representing five families of antimicrobial agents were performed. Plasmids carrying cephalosporin resistant (CR) genes were characterised. Sixty-six S. enterica isolates were recovered from five of eight farms. Forty-seven isolates were multi-AR and four contained bla CTX-M genes harboured in conjugative plasmids of the IncI1 family; three of these isolates were recovered before treatment with ceftiofur. The most frequent AR genes detected were tet(A) (51/66, 77%), sul1 (17/66, 26%); tet(B) (15/66, 23%) and qnrB (10/66, 15%). A direct relation between the use of ceftiofur in these conditions and the occurrence of CR S. enterica was not established. However, multi-AR was common, especially for ampicillin, streptomycin, sulphonamides and tetracycline. These antibiotics are used frequently in veterinary medicine in Spain and, therefore, should be used sparingly to minimise the spread of multi-AR. Copyright © 2018 The Authors. Published by Elsevier Ltd.. All rights reserved.
Full Text Available Abstract Parainfluenza virus is an important pathogen threatening the health of animals and human, which brings human many kinds of disease, especially lower respiratory tract infection involving infants and young children. In order to control the virus, it is necessary to fully understand the molecular basis resulting in the genetic diversity of the virus. Homologous recombination is one of mechanisms for the rapid change of genetic diversity. However, as a negative-strand virus, it is unknown whether the recombination can naturally take place in human PIV. In this study, we isolated and identified a mosaic serotype 3 human PIV (HPIV3 from in China, and also provided several putative PIV mosaics from previous reports to reveal that the recombination can naturally occur in the virus. In addition, two swine PIV3 isolates transferred from cattle to pigs were found to have mosaic genomes. These results suggest that homologous recombination can promote the genetic diversity and potentially bring some novel biologic characteristics of HPIV.
Toboldt, Anne; Tietze, Erhard; Helmuth, Reiner; Junker, Ernst; Fruth, Angelika
Salmonella enterica serovar 4,,12:b:− is a monophasic serovar not able to express the second-phase flagellar antigen (H2 antigen). In Germany, the serovar is occasionally isolated from poultry, reptiles, fish, food, and humans. In this study, a selection of 67 epidemiologically unrelated Salmonella enterica serovar 4,,12:b:− strains isolated in Germany between 2000 and 2011 from the environment, animal, food, and humans was investigated by phenotypic and genotypic methods to better understand the population structure and to identify potential sources of human infections. Strains of this monophasic serovar were highly diverse. Within the 67 strains analyzed, we identified 52 different pulsed-field gel electrophoresis XbaI profiles, 12 different multilocus sequence types (STs), and 18 different pathogenicity array types. The relatedness of strains based on the pathogenicity gene repertoire (102 markers tested) was in good agreement with grouping by MLST. S. enterica serovar 4,,12:b:− is distributed across multiple unrelated eBurst groups and consequently is highly polyphyletic. Two sequence types (ST88 and ST127) were linked to S. enterica serovar Paratyphi B (d-tartrate positive), two single-locus variants of ST1583 were linked to S. enterica serovar Abony, and one sequence type (ST1484) was associated with S. enterica serovar Mygdal, a recently defined, new serovar. From the characterization of clinical isolates and those of nonhuman origin, it can be concluded that the potential sources of sporadic human infections with S. enterica serovar 4,,12:b:− most likely are mushrooms, shellfish/fish, and poultry. PMID:23793625
Dundore-Arias, José Pablo; Groves, Russell L; Barak, Jeri D
Phytophagous insects can encounter Salmonella enterica on contaminated plant surfaces and transmit externally adhered and internalized bacteria on and among leaves. Excretion of ingested S. enterica by the leafhopper Macrosteles quadrilineatus has been previously reported; however, the sites of persistence of ingested bacteria remain undetermined. Fluorescence microscopy revealed the presence and persistence of S. enterica in various organs of M. quadrilineatus fed an inoculated diet for 12 h and then moved to two consecutive noninoculated diets for a total of 48 h. Ingested S. enterica was predominantly observed in the filter chamber, midgut, and Malpighian tubules of M. quadrilineatus dissected immediately after acquisition and at 24- and 48-h post-acquisition access periods (post-AAPs). Additionally, we examined the potential roles of the Salmonella pathogenicity island 1 (SPI-1) and SPI-2 type III secretion systems (T3SSs) in the persistence and excretion of ingested S. enterica. In competition assays, a prgH mutant lacking a functional SPI-1 T3SS was recovered at significantly lower levels than the WT in insect homogenates at 24 h post-AAP, and complementation with prgH restored S. enterica persistence in M. quadrilineatus. Moreover, expression of prgH inside M. quadrilineatus was observed up to 48 post-AAP. No differences were observed between the WT and an ssaK mutant lacking a functional SPI-2 T3SS in insect homogenates or between the WT and either mutant in insect excretions. This study provides novel insight into the presence and persistence of S. enterica inside M. quadrilineatus and demonstrates that the SPI-1 T3SS influences the persistence of the pathogen in the gut of a potential vector. Copyright © 2015, American Society for Microbiology. All Rights Reserved.
Zhou, Bin; Luo, Yaguang; Nou, Xiangwu; Yang, Yang; Wu, Yunpeng; Wang, Qin
Salmonella internalization in tomatoes during postharvest handling is a major food safety concern. This study was conducted to determine the effect of immersion time, immersion depth, and temperature differential between bacterial suspension and tomato pulp on the internalization of Salmonella enterica in tomato fruits. The effect of storage temperature and duration on the survival and growth of internalized Salmonella cells was also evaluated. Overall, immersion time significantly affected the incidence and extent of S. enterica internalization (P internalization. The depth of Salmonella internalization in tomato tissues also increased with increasing immersion time. Immersion time also significantly influenced the degree to which the temperature differential affected Salmonella internalization. With an immersion time of 2 min, the temperature differential had no significant effect on Salmonella internalization (P = 0.2536). However, with an immersion time of 15 min, a significantly larger Salmonella population became internalized in tomatoes immersed in solutions with a -30°F (-16.7°C) temperature differential. Internalized S. enterica cells persisted in the core tissues during 14 days of storage. Strain type and storage duration significantly affected (P internalized Salmonella recovered, but storage temperatures of 55 to 70°F (12.8 to 21.1°C) did not (P > 0.05). These findings indicate the importance of preventing pathogen internalization during postharvest handling.
Patchanee, Prapas; Boonkhot, Phacharaporn; Kittiwan, Nattinee; Tadee, Pakpoom; Chotinun, Suwit
Food-borne illness caused by Salmonella enterica remains a public health problem and results in economic loss worldwide. With the up-coming establish- ment of the ASEAN Economic Community (AEC) allowing unrestricted move- ment of labor and goods, there is a higher risk of pathogen transmission among the AEC countries. This study characterized and investigated the spatial and temporal associations of S. enterica strains isolated in AEC countries during 1940- 2012 compared with those isolated in northern-Thailand during 2011-2013. Of the 173 S. enterica strains examined, 68 sequence types (STs) and 32 clonal complexes (CCs) were identified by multi loci sequence typing. Twenty-one strains belonged to four sequence types new to AEC countries, and they constituted only two CCs. A number of strains originated from various countries with multiple hosts, were highlighted. There was evidence of strains circulating in the AEC region well over a decade. Such information will be important in formulating biosecurity measures, as well as in educating regarding the risk of disease transmission in AEC.
Howe, Kevin; Salehi, Sanaz; Hartford Bailey, R; Brooks, John P; Wills, Robert; Lawrence, Mark L; Karsi, Attila
Critical to the development of Salmonellosis in humans is the interaction of the bacterium with the epithelial lining of the gastrointestinal tract. Traditional scientific reasoning held type III secretion system (T3SS) as the virulence factor responsible for bacterial invasion. In this study, field-isolated Salmonella enterica serovar Kentucky and a known human pathogen Salmonella enterica serovar Typhimurium were mutated and evaluated for the invasion of human colorectal adenocarcinoma epithelial cells. S. enterica serovar Kentucky was shown to actively invade a eukaryotic monolayer, though at a rate that was significantly lower than Typhimurium. Additionally, strains mutated for T3SS formation were less invasive than the wild-type strains, but the decrease in invasion was not significant in Kentucky. Strains mutated for T3SS formation were able to initiate invasion of the eukaryotic monolayer to varying degrees based on strain, In the case of Kentucky, the mutated strain initiated invasion at a level that was not significantly different from the wild-type strain. A different result was observed for Typhimurium as the mutation significantly lowered the rate of invasion in comparison to the wild-type strain.
Andrew C. Elton
Full Text Available Salmonella meningitis is a rare manifestation of meningitis typically presenting in neonates and the elderly. This infection typically associates with foodborne outbreaks in developing nations and AIDS-endemic regions. We report a case of a 19-year-old male presenting with altered mental status after 3-day absence from work at a Wisconsin tourist area. He was febrile, tachycardic, and tachypneic with a GCS of 8. The patient was intubated and a presumptive diagnosis of meningitis was made. Treatment was initiated with ceftriaxone, vancomycin, acyclovir, dexamethasone, and fluid resuscitation. A lumbar puncture showed cloudy CSF with Gram negative rods. He was admitted to the ICU. CSF culture confirmed Salmonella enterica subsp. I (enterica Enteritidis (A. Based on this finding, a 4th-generation HIV antibody/p24 antigen test was sent. When this returned positive, a CD4 count was obtained and showed 3 cells/mm3, confirming AIDS. The patient ultimately received 38 days of ceftriaxone, was placed on elvitegravir, cobicistat, emtricitabine, and tenofovir alafenamide (Genvoya for HIV/AIDS, and was discharged neurologically intact after a 44-day admission.
Silva-Hidalgo, Gabriela; López-Valenzuela, Martin; Cárcamo-Aréchiga, Nora; Cota-Guajardo, Silvia; López-Salazar, Mayra; Montiel-Vázquez, Edith
bapA, previously named stm2689, encodes the BapA protein, which, along with cellulose and fimbriae, constitutes biofilms. Biofilms are communities of microorganisms that grow in a matrix of exopolysaccharides and may adhere to living tissues or inert surfaces. Biofilm formation is associated with the ability to persist in different environments, which contributes to the pathogenicity of several species. We analyzed the presence of bapA in 83 strains belonging to 17 serovars of Salmonella enterica subsp. enterica from wildlife in captivity at Culiacan's Zoo and Mazatlán's Aquarium. Each isolate amplified a product of 667 bp, which corresponds to the expected size of the bapA initiator, with no observed variation between different serovars analyzed. bapA gene was found to be highly conserved in Salmonella and can be targeted for the genus-specific detection of this organism from different sources. Since bapA expression improves bacterial proliferation outside of the host and facilitates resistance to disinfectants and desiccation, the survival of Salmonella in natural habitats may be favored. Thus, the risk of bacterial contamination from these animals is increased.
Internalisation potential of Escherichia coli O157:H7, Listeria monocytogenes, Salmonella enterica subsp. enterica serovar Typhimurium and Staphylococcus aureus in lettuce seedlings and mature plants.
Standing, Taryn-Ann; du Plessis, Erika; Duvenage, Stacey; Korsten, Lise
The internalisation potential of Listeria monocytogenes, Staphylococcus aureus, Escherichia coli O157:H7 and Salmonella enterica subsp. enterica serovar Typhimurium in lettuce was evaluated using seedlings grown in vermiculite in seedling trays as well as hydroponically grown lettuce. Sterile distilled water was spiked with one of the four human pathogenic bacteria (10(5) CFU/mL) and used to irrigate the plants. The potential for pathogen internalisation was investigated over time using light microscopy, transmission electron microscopy and viable plate counts. Additionally, the identities of the pathogens isolated from internal lettuce plant tissues were confirmed using polymerase chain reaction with pathogen-specific oligonucleotides. Internalisation of each of the human pathogens was evident in both lettuce seedlings and hydroponically grown mature lettuce plants. To our knowledge, this is the first report of S. aureus internalisation in lettuce plants. In addition, the levels of background microflora in the lettuce plants were determined by plate counting and the isolates identified using matrix-assisted laser ionisation-time of flight (MALDI-TOF). Background microflora assessments confirmed the absence of the four pathogens evaluated in this study. A low titre of previously described endophytes and soil inhabitants, i.e., Enterobacter cloacae, Enterococcus faecalis, Lysinibacillus fusiformis, Rhodococcus rhodochrous, Staphylococcus epidermidis and Staphylococcus hominis were identified.
Resistance phenotypes and genotypes among multiple-antimicrobial-resistant Salmonella enterica subspecies enterica serovar Choleraesuis strains isolated between 2008 and 2012 from slaughter pigs in Okinawa Prefecture, Japan.
Matayoshi, Masanao; Kitano, Takashi; Sasaki, Tetsu; Nakamura, Masaji
A total of 349 Salmonella enterica subspecies enterica serovar Choleraesuis (S. Choleraesuis) strains, which were isolated between 2008 and 2012 from 349 pigs at two slaughterhouses in Okinawa Prefecture, Japan, were investigated for antimicrobial susceptibility and the presence of antimicrobial resistance genes. All isolates were resistant to at least four antimicrobial agents. The antimicrobial agents for which isolates showed a high incidence of resistance were as follows: ampicillin (100%) and streptomycin (100%), followed by gentamicin (99.7%), oxytetracycline (99.7%), sulfamethoxazole/trimethoprim (99.4%), nalidixic acid (40.1%) and oxolinic acid (40.1%). All isolates were sensitive to cefuroxime, ceftiofur, colistin, fosfomycin, enrofloxacin, orbifloxacin and danofloxacin. The predominant resistance phenotypes and genotypes were: resistance to ampicillin, streptomycin, gentamicin, oxytetracycline and sulfamethoxazole/trimethoprim (58.5%, 204/349) and blaTEM-strA-strB-aadA1-aadA2-aacC2-tet (B)-sul1-sul2-dhfrXII-dhfrXIII (36.1%, 126/349). The quinolone resistance-determining regions (QRDRs) of gyrA, gyrB, parC and parE of the quinolone-resistant isolates (n=12) showed amino acid substitutions of Ser-83→Phe or Asp-87→Tyr in GyrA and Ser-107→Ala in ParC. To our knowledge, this is the first report on the molecular characterization of antimicrobial resistance among S. Choleraesuis strains in Japan.
inhibit other BoNT serotypes or thermolysin . 15. SUBJECT TERMS Clostridium botulinum, neurotoxin, serotype A, peptidomimetic inhibitor , light chain...20 0 BoNT/E SNAP-25, residues 1–206 20 0 BoNT/F VAMP residues 1–94 20 0 Thermolysin Synthetic peptidea 500 0 The inhibitor concentration was 4 mM. a...Structure ArticleA Potent Peptidomimetic Inhibitor of Botulinum Neurotoxin Serotype A Has a Very Different Conformation than SNAP-25 Substrate Jorge
Méndez-Lage, Susana; Losada-Castillo, Isabel; Agulla-Budiño, Andrés
To examine the epidemiology of pneumococcal infection in Galicia (Spain) after the incorporation of the pneumococcal conjugate vaccine, and to determine serotype distribution, antibiotic susceptibility, risk factors and associated mortality in cases of invasive pneumococcal disease (IPD) during 2011 and 2012. All strains causing IPD in Galicia were studied. Serotyping was performed by agglutination and Quellung reaction. Antibiotic sensitivity to penicillin, cefotaxime, erythromycin, vancomycin, and levofloxacin was determined. The risk factors considered were chronic respiratory disease, heart disease, liver disease, kidney disease, diabetes mellitus, and HIV and non-HIV immunodeficiency. A total of 555 strains were collected, with 43 different serotypes being found. The most frequently isolated ones were: serotype3 (17.5%), serotype7F (12.6%), serotype19A (9.4%), serotype14 (4.1%), serotype6C (4.1%), serotype11A (4%) and serotype22F (3.8%). 57.1% of isolates were serotypes included in VNC-13V. Two non-penicillin-sensitive strains and two others were not sensitive to cefotaxime, and 24.7% of the strains were not susceptible to erythromycin (26.9% in 2011 and 22.5% in 2012). The case fatality rate was 16.5%, reaching 23.3% in patients over 75years. Diseases with a statistically significant risk of mortality were: liver, kidney and immunodeficiency without HIV. Serotype3 was the most frequent in Galicia. Very few strains were not susceptible to penicillin. Erythromycin resistance decreased from 2011 to 2012. It is highlighted that mortality increases with age. Liver disease, renal disease and non-HIV immunodeficiency increases the mortality risk. Copyright © 2015 Elsevier España, S.L.U. y Sociedad Española de Enfermedades Infecciosas y Microbiología Clínica. All rights reserved.
Kim, Han Wool; Lee, Ji Hyen; Cho, Hye Kyung; Lee, Hyunju; Seo, Ho Seong; Lee, Soyoung; Kim, Kyung Hyo
Group B streptococcus (GBS) infection is a leading cause of sepsis and meningitis among infants, and is associated with high rates of morbidity and mortality in many countries. Protection against GBS typically involves antibody-mediated opsonization by phagocytes and complement components. The present study evaluated serotype-specific functional antibodies to GBS among Korean infants and in intravenous immunoglobulin (IVIG) products. An opsonophagocytic killing assay (OPA) was used to calculate the opsonization indices (OIs) of functional antibodies to serotypes Ia, Ib, and III in 19 IVIG products from 5 international manufacturers and among 98 Korean infants (age: 0-11 months). The GBS Ia, Ib, and III serotypes were selected because they are included in a trivalent GBS vaccine formulation that is being developed. The OI values for the IVIG products were 635-5,706 (serotype Ia), 488-1,421 (serotype Ib), and 962-3,315 (serotype III), and none of the IVIG lots exhibited undetectable OI values (Korean manufacturers. The seropositive rate among infants was significantly lower for serotype Ia (18.4%), compared to serotype Ib and serotype III (both, 38.8%). Infant age of ≥ 3 months was positively correlated with the seropositive rates for each serotype. Therefore, only a limited proportion of infants exhibited protective immunity against serotype Ia, Ib, and III GBS infections. IVIG products that exhibit high antibody titers may be a useful therapeutic or preventive measure for infants. Further studies are needed to evaluate additional serotypes and age groups. © 2017 The Korean Academy of Medical Sciences.
Wang, Hsi-Kai; Tsai, Cheng-Han; Chen, Kuan-Hung; Tang, Chung-Tao; Leou, Jiun-Shyang; Li, Pi-Chun; Tang, Yin-Liang; Hsieh, Hsyue-Jen; Wu, Han-Chung; Cheng, Chao-Min
Here, two types of cellulose-based in vitro diagnostic devices are demonstrated for the diagnosis of dengue virus infection in both buffer system and human serum: 1) paper-based ELISA for providing the semiquantitative information of the disease activity of serotype-2 dengue fever to healthcare persons (i.e., monitoring the disease activity with a specific serotype in single patients); 2) lateral flow immunoassays to screen for infection with serotype-2 dengue fever (i.e., rapid YES or NO diagnosis prepared for large populations, in terms of global public health). Paper-based ELISA (specific to serotype-2 dengue fever), which builds off of our previous studies and a revised previous ELISA procedure, owns multiple advantages: 1) high sensitivity (about 40 times higher than the current ELISA-based approaches, due to our therapeutic-based monoclonal antibody) and specificity (specific to dengue virus serotype-2 nonstructural protein-1 antigens); 2) tiny amount of sample and reagent used for single tests; 3) short operating duration (i.e., rapid diagnostic device); and, 4) inexpensiveness (appropriate for use in all developing and underdeveloped nations of the world). Due to the higher sensitivity and shorter operating duration of paper-based ELISA (compared with conventional ELISA, and lateral flow immunoassays also performed in this study), this study has not only been able to perform the diagnosis of dengue virus serotype-2 nonstructural protein-1 antigens in both buffer system and human serum but also to evaluate dengue virus serotype-2 envelope proteins in the buffer system, thus successfully achieving the first such use of these proteins as the target antigen for the development of diagnostic tools. These results provide a more comprehensive understanding for the genesis of dengue fever diagnostic tools (through antibody-antigen recognition). Copyright © 2014 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.
Janoir, Claire; Cohen, Robert; Levy, Corinne; Bingen, Edouard; Lepoutre, Agnès; Gutmann, Laurent; Varon, Emmanuelle
In France, the use of the 7-valent pneumococcal conjugate vaccine (PCV7) lead to an overall significant decrease in PCV7 invasive pneumococcal disease (IPD) incidence. However, the decrease in vaccine serotype prevalence was partially counterbalanced by the serotype replacement phenomenon. In this study, we analyzed the role of the newly described serotype 6C as one of the replacement serotypes. This work was conducted on a large time scale from the early PCV7 era (2002–2003) to the PCV13 era (2010–2011), both on IPD strains recovered from the whole population and nasopharyngeal colonizing strains isolated in infant less than two years, who are known to be the main reservoir for pneumococci. Serotype 6C took advantage over 6A and 6B serotypes, which both decreased over time. A continuous and significant increase in 6C IPD was observed in adults along the study period; in contrast, in children less than two years, only an increase in 6C nasopharyngeal carriage was found, the prevalence of serotype 6C in IPD remaining very low over time. Among 101 6C invasive and colonizing strains studied by MLST, 24 STs were found to be related to three major clonal complexes, CC395, CC176, and CC315. STs related to CC176 tend to disappear after 2009 and were essentially replaced by ST386 (CC315), which dramatically increased over time. This clonal expansion may be explained by the erythromycin and tetracycline resistances associated with this clone. Finally, the decrease observed in nasopharyngeal 6C carriage since 2010, likely related to the PCV13 introduction in the French immunization schedule, is expected to lead to a decrease in 6C IPD in adults thereafter. PMID:24603763
Full Text Available There is a paucity of longitudinal data on the serotype-specific burden of invasive group B Streptococcus (GBS disease from low-middle income countries, which could inform selection of vaccine epitopes.From 2005 to 2014, infants less than 90 days of age with invasive GBS disease were identified through sentinel laboratory and hospital admission surveillance at Chris Hani Baragwanath Academic Hospital in Soweto, South Africa.We identified 820 cases of invasive GBS disease, including 55% among newborns <7 days age (i.e. early-onset disease; EOD. The overall incidence (per 1,000 live births of invasive GBS disease was 2.59 (95% CI: 2.42-2.77, including 1.41 (95% CI: 1.28-1.55 for EOD and 1.18 (95% CI: 1.06-1.30 in infants 7-89 days age (late-onset disease. Year-on-year, from 2005 to 2014, we observed a 9.4% increase in incidence of serotype Ia invasive disease (RR: 1.09; 95% CI: 1.04-1.15; p<0.001, and a 7.4% decline in serotype III invasive disease (RR: 0.93; 95% CI: 0.90-0.96; p<0.001. Overall, serotypes Ia (28.2%, III (55.4% and V (7.9% were the commonest disease causing serotypes.The incidence of invasive GBS disease has remained persistently high in our setting, with some changes in serotype distribution, albeit mainly involving the same group of dominant serotypes.
Han, Sanghyun; Micallef, Shirley A
Foodborne illness-causing enteric bacteria are able to colonize plant surfaces without causing infection. We lack an understanding of how epiphytic persistence of enteric bacteria occurs on plants, possibly as an adaptive transit strategy to maximize chances of reentering herbivorous hosts. We used tomato (Solanum lycopersicum) cultivars that have exhibited differential susceptibilities to Salmonella enterica colonization to investigate the influence of plant surface compounds and exudates on enteric bacterial populations. Tomato fruit, shoot, and root exudates collected at different developmental stages supported growth of S. enterica to various degrees in a cultivar- and plant organ-dependent manner. S. enterica growth in fruit exudates of various cultivars correlated with epiphytic growth data (R(2) = 0.504; P = 0.006), providing evidence that plant surface compounds drive bacterial colonization success. Chemical profiling of tomato surface compounds with gas chromatography-time of flight mass spectrometry (GC-TOF-MS) provided valuable information about the metabolic environment on fruit, shoot, and root surfaces. Hierarchical cluster analysis of the data revealed quantitative differences in phytocompounds among cultivars and changes over a developmental course and by plant organ (P enterica growth, while fatty acids, including palmitic and oleic acids, were negatively correlated. We demonstrate that the plant surface metabolite landscape has a significant impact on S. enterica growth and colonization efficiency. This environmental metabolomics approach provides an avenue to understand interactions between human pathogens and plants that could lead to strategies to identify or breed crop cultivars for microbiologically safer produce. In recent years, fresh produce has emerged as a leading food vehicle for enteric pathogens. Salmonella-contaminated tomatoes represent a recurrent human pathogen-plant commodity pair. We demonstrate that Salmonella can utilize
Full Text Available There are no licensed vaccines available against Moraxella catarrhalis, a significant human respiratory pathogen. Lipooligosaccharide (LOS based conjugate vaccines derived from individual serotype M. catarrhalis only showed partial protection coverage. A vaccine combining LOS conjugates of two or three serotypes might provide a broader protection.Mice were immunized intranasally with the combined conjugates consisting of LOS from serotype A and B or serotype A, B, and C followed by challenge with different M. catarrhalis strains of three serotypes. Mouse lungs, nasal washes, and sera were collected after each challenge for bacterial counts, histological evaluation, cytokine profiles, antibody level and binding activity determinations.Intranasal administration of the combined LOS conjugates not only enhanced pulmonary bacterial clearance of all three serotypes of M. catarrhalis strains in vaccinated mice, but also elevated serotype-specific anti-LOS immunoglobulin (IgA and IgG titers in nasal wash and serum respectively. Mice vaccinated with the combined LOS conjugates also showed increased interferon (IFN-γ, interleukin (IL-12, and IL-4 in the lungs after challenges. Compared to the control group, mice immunized with the combined LOS conjugates also showed reduced lung inflammation after M. catarrhalis infections. The hyperimmune sera induced by the combined conjugates exhibited a broad cross-reactivity toward all three serotypes of M. catarrhalis under transmission electron microscopy.The combined vaccine of serotype A and B LOS conjugates provides protection against most M. catarrhalis strains by eliciting humoral and cellular immune responses.
Conclusion: Serotype distribution, antimicrobial susceptibility, and sequence type characterization in Asia and in other global regions may contribute to improve the prevention and treatment of neonatal GBS infections.
Calayag, Alyzza Marie B; Paclibare, Phyllis Anne P; Santos, Pauline Dianne M; Bautista, Corinne Aimee C; Rivera, Windell L
Salmonella enterica is a well-known pathogen commonly acquired from the consumption of contaminated food. It has been estimated to affect millions of humans and cause hundreds of thousands of deaths per year globally. Pork, one of the most commonly consumed meats worldwide, has been identified as one of the main sources of human salmonellosis. In this study, we aimed to detect and characterize S. enterica from slaughtered swine and generate antimicrobial resistance profiles of select isolates. Tonsils and jejunum with mesenteric lymph nodes (MLN) were collected from a total of 240 swine from eight abattoirs (five accredited and three locally registered abattoirs) across Metro Manila. S. enterica were isolated using conventional culture methods and confirmed by PCR amplification of the invA gene. Isolates were further characterized based on somatic antigen by multiplex PCR. We report that there is no significant difference (P = 0.42) between the incidences of S. enterica in swine slaughtered in accredited (44.0%) and in locally registered abattoirs (46.7%). Most samples were contaminated with S. enterica under serogroup O:3,10. Antimicrobial susceptibility testing of 183 isolates using the VITEK ® 2 system revealed high resistance to ampicillin (67.8%) and trimethoprim/sulfamethoxazole (80.3%). Multidrug-resistance was found in 124 (67.8%) isolates. Copyright © 2017 Elsevier Ltd. All rights reserved.
Kim, Yoonbin; Kim, Hoikyung; Beuchat, Larry R; Ryu, Jee-Hoon
Non-pathogenic bacterial biofilms were developed on the surface of stainless steel possessing desiccation tolerance and antimicrobial activity against Salmonella enterica. Three bacteria exhibiting strong antimicrobial activities against S. enterica were isolated from various soils, foods, and food-contact surfaces. Isolates were identified as Pseudomonas extremorientalis (strain Lettuce-28), Paenibacillus peoriae (strain Lettuce-7), and Streptomyces cirratus (strain Geumsan-207). These bacteria grew rapidly and formed biofilms within 24 h on the surface of stainless steel coupons (SSCs) immersed in laboratory media (tryptic soy broth or Bennet's broth) at 25 °C. Cells in biofilms had enhanced tolerance to desiccation (exposure to 43% atmospheric relative humidity [RH]) and retained antimicrobial activity against S. enterica. Populations of S. enterica deposited on SSCs containing biofilm formed by Ps. extremorientalis strain Lettuce-28, for example, decreased by > 2.5 log CFU/coupon within 24 h at 25 °C and 43% RH, while the number of cells inoculated on SSCs lacking biofilm decreased by 1.5 log CFU/coupon. Antimicrobial activities of the three antagonistic bacteria against S. enterica persisted in desiccated biofilms. This study provides insights to developing strategies to inactivate Salmonella and perhaps other foodborne pathogens on abiotic surfaces using non-pathogenic antagonistic bacteria. Copyright © 2017. Published by Elsevier Ltd.
Gao, Qingqing; Zhang, Debao; Ye, Zhengqin; Zhu, Xiaoping; Yang, Weixia; Dong, Lanmei; Gao, Song; Liu, Xiufan
Urinary tract infections (UTIs) are among the most common human diseases worldwide. This study aimed to collect uropathogenic Escherichia coli (UPEC) isolates from Jiangsu Province and obtain insights into the molecular epidemiology of UPEC in this region. The O serotypes, phylogenetic groups, and virulence factors of 183 UPEC isolates were determined. In this study, we isolated 51 UPEC isolates with common O serotypes including O1, O2, O4, O6, O7, O16, O18 and O75, as well as 35 of those with uncommonly encountered O serotypes including O8, O12, O15, O26, and O74. Groups B2 and D were the most prevalent phylogenetic groups and accounted for 29.5% and 41% of the isolates, respectively. In the tested 13 virulence genes (VGs), tonB and dsdA possessed the highest prevalence rate, followed by fimH, degP and ompR. Several other virulence genes such as fliC, neuC, ireA, and vat had prevalence less than 23%. Moreover, representative isolates belonging to common or uncommon O serotypes with different numbers of VGs were chosen for the pathogenic analyses. Based on the results of 1-day-old chick lethality assay and UTI ascending mouse infection model, our study suggested that the virulence of UPEC isolates for chicks and/or mice depended on both the number of VGs expressed and the O serotypes. Copyright © 2017 Elsevier Ltd. All rights reserved.
Nan Nwe Win
Full Text Available Determination of hepatitis C virus (HCV genotypes plays an important role in the direct-acting agent era. Discrepancies between HCV genotyping and serotyping assays are occasionally observed. Eighteen samples with discrepant results between genotyping and serotyping methods were analyzed. HCV serotyping and genotyping were based on the HCV nonstructural 4 (NS4 region and 5′-untranslated region (5′-UTR, respectively. HCV core and NS4 regions were chosen to be sequenced and were compared with the genotyping and serotyping results. Deep sequencing was also performed for the corresponding HCV NS4 regions. Seventeen out of 18 discrepant samples could be sequenced by the Sanger method. Both HCV core and NS4 sequences were concordant with that of genotyping in the 5′-UTR in all 17 samples. In cloning analysis of the HCV NS4 region, there were several amino acid variations, but each sequence was much closer to the peptide with the same genotype. Deep sequencing revealed that minor clones with different subgenotypes existed in two of the 17 samples. Genotyping by genome amplification showed high consistency, while several false reactions were detected by serotyping. The deep sequencing method also provides accurate genotyping results and may be useful for analyzing discrepant cases. HCV genotyping should be correctly determined before antiviral treatment.
Full Text Available Cryptococcus neoformans is a major human pathogenic fungus that can cause meningoencephalitis in immunocompromised hosts. It contains two divergent varieties, var. grubii (serotype A and var. neoformans (serotype D, as well as hybrids (serotype AD between these two varieties. In this study, we investigated the extent of chromosomal rearrangements between the two varieties, estimated the effects of chromosomal rearrangements on recombination frequencies, and surveyed the potential polymorphisms of the rearrangements among natural strains of the three serotypes. Through the analyses of two sequenced genomes from strains H99 (representing var. grubii and JEC21 (representing var. neoformans, we revealed a total of 32 unambiguous chromosome rearrangements, including five translocations, nine simple inversions, and 18 complex rearrangements. Our analyses identified that overall, rearranged regions had recombination frequencies about half of those around syntenic regions. Using a direct PCR screening strategy, we examined the potential polymorphisms of 11 rearrangements among 64 natural C. neoformans strains from five countries. We found no polymorphism within var. neoformans and very limited polymorphism within var. grubii. However, strains of serotype AD showed significant polymorphism, consistent with their hybrid origins coupled with differential loss of heterozygosity. We discuss the implications of these results on the genome structure, ecology, and evolution of C. neoformans.
Parra, Eliana L.; Ramos, Viviana; Sanabria, Olga; Moreno, Jaime
In Colombia, a laboratory-based surveillance of invasive Streptococcus pneumoniae isolates as part of SIREVA II PAHO has been conducted since 1994. This study describes the serotype distribution, antimicrobial resistance, and genetic relationships of pneumococcal isolates recovered in Colombia from 2005 to 2010. In this study, demographic data of invasive S. pneumoniae isolates were analyzed, and antimicrobial susceptibility patterns were determined. Pulse field gel electrophoresis (n = 629) and multilocus sequence typing (n = 10) were used to determine genetic relationship of isolates with minimal inhibitory concentration to penicillin ≥0.125 µg/mL. A total of 1775 isolates of S. pneumoniae were obtained. Fifteen serotypes accounted for 80.7% of isolates. Serotype 14 (23.1%) was the most frequent in the general population. Penicillin resistance was 30.7% in meningitis and 9.0% in non-meningitis. Clones Spain6BST90, Spain9VST156, Spain23FST81, and Colombia23FST338 were associated to isolates. Additionally, serotype 6A isolates were associated with ST460 and ST473, and 19A isolates with ST276, ST320, and ST1118. In conclusion, the surveillance program provided updated information of trends in serotype distribution, antimicrobial resistance and the circulation of clones in invasive pneumococcal diseases. These results could be helpful to understand the epidemiology of S. pneumoniae in Colombia, and provide a baseline to measure the impact of vaccine introduction. PMID:24416330
Eva Del Amo
Full Text Available The aim of this study was to study the serotypes and clonal diversity of pneumococci causing invasive pneumococcal disease in Catalonia, Spain, in the era of 13-valent pneumococcal conjugate vaccine (PCV13. In our region, this vaccine is only available in the private market and it is estimated a PCV13 vaccine coverage around 55% in children. A total of 1551 pneumococcal invasive isolates received between 2010 and 2013 in the Molecular Microbiology Department at Hospital Sant Joan de Déu, Barcelona, were included. Fifty-two serotypes and 249 clonal types-defined by MLST-were identified. The most common serotypes were serotype 1 (n = 182; 11.7%, 3 (n = 145; 9.3%, 19A (n = 137; 8.8% and 7F (n = 122; 7.9%. Serotype 14 was the third most frequent serotype in children < 2 years (15 of 159 isolates. PCV7 serotypes maintained their proportion along the period of study, 16.6% in 2010 to 13.4% in 2013, whereas there was a significant proportional decrease in PCV13 serotypes, 65.3% in 2010 to 48.9% in 2013 (p<0.01. This decrease was mainly attributable to serotypes 19A and 7F. Serotype 12F achieved the third position in 2013 (n = 22, 6.4%. The most frequent clonal types found were ST306 (n = 154, 9.9%, ST191 (n = 111, 7.2%, ST989 (n = 85, 5.5% and ST180 (n = 80, 5.2%. Despite their decrease, PCV13 serotypes continue to be a major cause of disease in Spain. These results emphasize the need for complete PCV13 vaccination.