Non-typhoidal Salmonella Isolates Research Articles

Emergent multidrug-resistant nontyphoidal Salmonella serovars isolated from poultry in Brazil coharboring blaCTX-M-2 and qnrB or blaCMY-2 in large plasmids

The number of foodborne gastroenteritis caused by nontyphoidal Salmonella (NTS) worldwide is estimated to be 80.3 million each year. Currently, antimicrobial-resistant NTS disseminated in the animal environment increases the risk of aggravated foodborne outbreaks. Poultry are important source of foodborne NTS infections. This study was conducted to evaluate the phenotypic and genotypic characteristics of 83 NTS isolates from poultry, classified within 36 different serovars. The most prevalent serovar was S. Schwarzengrund (10/83), from which 8/10 were multidrug resistant (MDR). The antimicrobial susceptibility testing showed a total of 18 MDR isolates, from which 8/18 coharbored blaCTX-M-2 and qnrB5. The genes qnrB5, blaCTX-M-2, qnrB2, or blaCMY-2 were also found alone in other MDR isolates. All resistance genes were harbored in large plasmids, ranging from 30 to 270 kb. The pColE replicon was present in 8 MDR isolates; however it was not associated with resistance. ISCR1 and class I integron structures were always associated with blaCTX-M-2.

Poultry: A receptacle for non-typhoidal Salmonellae and antimicrobial resistance

Non-typhoidal Salmonellosis, a zoonotic infection associated with acute gastroenteritis is caused by non-typhoidal salmonellae (NTS). The study was carried out to determine the prevalence of NTS serovars and their antimicrobial resistance along with the presence of the virulence gene (invA gene) in poultry samples. This is a prospective cross-sectional study carried out at the Enteric Diseases Division, Kasturba Medical College, Manipal, South India from January 2016- December 2017. Poultry samples were collected randomly from two local poultry farms in Udupi district and processed following CDC standard protocol. From the 396 poultry meat samples, intestinal contents and faecal samples collected, 58 NTS serovars were isolated showing a prevalence of 14.64%. Salmonella Infantis, 43.1%, 25/58 was the commonest serovar. Resistance to ciprofloxacin 72.41%, ampicillin 32.8%, gentamicin 17.24%, cotrimoxazole 29.31% and amoxicillin-clavulanic acid 6.9% was observed. The invA gene was detected in 43 NTS isolates (74.13%). Poultry sources are recognized as a significant cause for non-typhoidal salmonellosis. Therefore, hygienic measures should be initiated to reduce the contamination of meat and poultry products with virulent strains of Salmonella that are of public health significance.

Nontyphoid Salmonella carriage, serovar profile and antimicrobial resistance phenotypes in slaughter cattle

AbstractCurrent nontyphoid Salmonella (NTS) carriage in 200 apparently healthy slaughter cattle by ISO 6579 standard bacteriology (ISO) was 1% (2/200) in carcass and fecal content, and 2% (4/200) in mesenterial lymph nodes. There was no isolation from liver, kidney, spleen, and gallbladder, with an overall prevalence of 4% (8/200). Real‐time PCR was in substantial agreement to ISO in confirming Salmonella‐suspect isolates (Relative Trueness: 93.33%). Predominant serovar was S. Enteritidis (50%) followed by S. Typhimurium (37.5%), and S. Albany (12.5%). Five and three of eight NTS isolates were susceptible (62.5%) and resistant (37.5%) to 18 antimicrobials, respectively. Only three S. Enteritidis isolates (37.5%) showed multidrug resistance to 2–3 of 7 antimicrobials (amikacin, cefotaxime, cefoxitin, gentamicin, norfloxacin, pefloxacin, and tobramycin). S. Enteritidis predominance over S. Typhimurium, first detection of S. Albany in cattle in Turkey, and sole resistance in mesenterial lymph node S. Enteritidis isolates highlights study findings.Practical applicationsContaminated carcass and related material, for example, fecal content and mesenterial lymph nodes of apparently healthy slaughter cattle carrying nontyphoid Salmonella serovars still pose significant health risk to public in Turkey, where bovine meat covers the highest annual red meat consumption quota with high demand to edible offal. In this study, current predominance of S. Enteritidis, particularly in mesenterial lymph nodes, and the MDR pattern identified; the presence of S. Typhimurium as the second dominant and pansusceptible serovar; detection of S. Albany for the first time in cattle fecal content are new epidemiological findings. This data could be used in revising both bovine meat and offal's actual NTS status, and the control and prevention programs in our country and in the neighboring countries of interest.

Carriage and antimicrobial resistance of non-typhoidal <i>Salmonella</i> in cattle slaughtered in Ambo municipality abattoir, West Shewa zone, Oromia, Ethiopia - a point prevalence survey

Cattle are reservoir for non-typhoidal Salmonella (NTS) serovars which are important foodborne zoonosis worldwide. A cross-sectional study was undertaken from October, 2015 to May, 2016 to estimate the prevalence and assess antimicrobial susceptibility profiles of NTS isolated from slaughtered cattle following standard microbiological technique. A total of 300 samples (150 mesenteric lymph nodes and 150 feces) from 150 cattle slaughtered at Ambo municipality abattoir were investigated. Standard disc-diffusion method was usedfor antimicrobial susceptibility testing of the isolates. Chi-square and Fisher’s exact tests were used to assess association between NTS prevalence and potential risk factors. An overall NTS prevalence of 8% (95 % confidence interval [CI]: 4.2-13.6%) was found at animal level. At sample level, NTS prevalence of 5.6% (17/300) (95% CI: 3.3-8.9%) was found. Prevalence of NTS in mesenteric lymph nodes and fecal samples was 6.6% and 4.6%, respectively. There was no significant difference in prevalence of NTS with respect to sex, age groups and body condition (p> 0.05). However, significantly higher prevalence was seen in animals from Guder market (12.5%) than Ambo market (1.6%) (p< 0.05). The highest level of antimicrobial resistance was observed for amoxicillin clavulanic acid (100%) and ampicillin (100%) followed by streptomycin (41.1%). Six of the 17 NTS isolates (35.3%) showed multi-drug resistance (MDR) to three or more antimicrobial classes. In conclusion, high percentage of NTS isolates were MDR posing public health threat to consumers. Improved hygiene, education, prudent use of antimicrobials and further large scale epidemiological studies are suggested.Keywords: Antimicrobial resistance; Cattle; Ethiopia; Non-typhoidal Salmonella;Prevalence

Molecular epidemiology of the emerging ceftriaxone resistant non-typhoidal Salmonella in southern Taiwan.

The increasing trend of ceftriaxone resistant non-typhoidal Salmonella (NTS) worldwide is of serious concern, however, data is lacked in southern Taiwan. Salmonella isolates were collected at a regional hospital in Kaohsiung during 2004-2013. Ceftriaxone resistant NTS isolates were further characterized for beta-lactamases, typed by pulsed field gel electrophoresis (PFGE), multilocus sequence typing (MLST) and their plasmids were analyzed by PCR replicon typing and plasmid mutilocus sequence typing. Among 528 NTS isolates, the most common serogroup is serogroup B (44.9%), followed by serogroup D, and serogroup C. Eleven (2.1%) isolates were resistant to ceftriaxone and were distributed in three peak periods (2010, 2011, and 2013). PFGE and MLST revealed the ten serogroup B isolates were of two clones. Beta-lactamase genes were detected in 10 of the 11 isolates, including CMY-2 (5 isolates), TEM-1 (2), CTX-M-14 (1), and 2 isolates carried both TEM-1 and CMY-2. Plasmid incompatibility types were identified in 9 (81.8%) isolates; three were IncI1, three was IncHI2, one was IncFIB and two had both replicons of IncI1 and IncHI2. The only ESBL gene blaCTM-X-14 was found in an isolate with plasmid belonged to IncHI2, which has not been reported in NTS in Taiwan before. Most MLST types and plasmid MLST types of NTS isolates in this study are different from those in northern Taiwan. Though clonal spread of ceftriaxone resistant NTS was suggested by PFGE and MLST, plasmid characterization and beta-lactamase detection revealed their plasmid types and beta-lactamase types were different.

Prevalence and antibiogram of non-typhoidal salmonella isolates from poultry in Ilorin, Kwara State

Prevalence and antibiogram of non-typhoidal salmonella isolates from poultry in Ilorin, Kwara State

Changes in epidemiology and antimicrobial susceptibility of nontyphoid Salmonella in children in southern Taiwan, 1997-2016.

This study aimed to investigate the changing trends in serogroup distribution and antimicrobial susceptibility of nontyphoid Salmonella (NTS) isolated from children in the past two decades in Taiwan. This retrospective study enrolled patients aged younger than 18 years with culture-proven NTS infection in a tertiary medical center from 1997 to 2016. Data on age, Salmonella serogroup, culture sample, and antimicrobial susceptibility were obtained from medical records. The rates of Salmonella serogroups and antimicrobial resistance were compared between period 1 (1997-2006) and period 2 (2007-2016). A total of 2075 NTS isolates were identified from 1997 to 2016 (1036 in period 1, 1039 in period 2). The major isolates were from feces (89.1%) and blood (10.4%). The most common serogroup was serogroup B (54.8%), followed by serogroup D (18.8%). There was a significant decline in the proportion of serogroup B with a concomitant rise in serogroup D in the past two decades. In period 2, resistance to cefotaxime and cefixime increased remarkably among serogroup B, whereas resistance to ampicillin increased notably among serogroup D. Furthermore, the incidence of multidrug-resistant (≥3 antibiotics) NTS significantly decreased in both serogroup B and serogroup D in period 2. This study revealed a significant rise in serogroup D with a concomitant decline in serogroup B NTS infection in Taiwanese children. Moreover, antimicrobial resistance to third-generation cephalosporins increased in serogroup B. Continuous surveillance of NTS serogroup distribution and antimicrobial susceptibility is mandatory to formulate therapeutic strategies for NTS infections.

Prevalence and antimicrobial resistance of non-typhoid Salmonella in military personnel, 1988-2013

Objective: To describe the spanning 25 years data for the occurrence, magnitude, and trends regarding antimicrobial resistance of non-typhoidal Salmonella (NTS) isolated from non-immune travelers to Thailand participating in joint military operations. Methods: A total of 355 NTS isolates, obtained from 2 052 fecal samples from US soldiers deployed for military maneuvers in Thailand during 1988-2013, were examined for NTS serogroup/ serotypes and tested for antimicrobial susceptibility by disk diffusion to these 10 antibiotics: ampicillin, azithromycin (AZM), ciprofloxacin, colistin, gentamicin, kanamycin, nalidixic acid, streptomycin (STR), tetracycline (TET), and trimethoprim/sulfamethoxazole. Identified AZM-resistant NTS isolates were further evaluated for their minimal inhibitory concentration by the E-test method. Results: NTS infections accounted for 17.3% (355/2 052), including 11 serogroups and 50 different serotypes. The most prevalent serogroup was Salmonella group C2-C3 (35.8%, 127/355) followed by groups B (21.1%, 75/355) and C1 (18.6%, 66/355). Identified serotypes included Salmonella hadar (n=60), Salmonella rissen (n=45), and Salmonella blockley (n=34). Among the predominate serogroups, antimicrobial resistance was consistently high against TET (76.9%, 273/355) followed by STR (40.8%, 145/355). One Salmonella senftenberg isolate demonstrated decreased ciprofloxacin susceptibility. Most isolates (94.6%) were resistant to one or more antimicrobials, and the most common multidrug resistance was TET-STR-nalidixic acid (11.5%, 41/355). Conclusions: The prevalence of NTS serotypes and the growing magnitude of antibiotic resistant bacteria isolated from deployed US military in Thailand are documented from 1988-2013. This study demonstrates the antibiotic resistance profiles, highlighting the effectiveness of AZM that is a first-line treatment for travelers to Southeast Asia. AZM-resistant NTS isolates are periodically observed over a 25- year period. Hence, the ongoing surveillance and prevalence efforts are required to monitor NTS resistant strains causing further treatment failure.

Antimicrobial residues and resistance against critically important antimicrobials in non-typhoidal Salmonella from meat sold at wet markets and supermarkets in Vietnam

Excessive antimicrobial usage and deficiencies in hygiene in meat production systems may result in undesirable human health hazards, such as the presence of antimicrobial drug residues and non-typhoidal Salmonella (NTS), including antimicrobial resistant (AMR) NTS. Recently, Vietnam has witnessed the emergence of integrated intensive animal production systems, coexisting with more traditional, locally-sourced wet markets. To date no systematic studies have been carried out to compare health hazards in beef, pork and chicken in different production systems. We aimed to: (1) estimate the prevalence of antimicrobial residues in beef, pork and chicken meat; (2) investigate the prevalence and levels of NTS contamination; and (3) investigate serovar distribution and AMR against critically important antimicrobials by animal species and type of retail (wet market vs. supermarket) in Vietnam. Fresh pork, beef and chicken meat samples (N=357) sourced from wet markets and supermarkets in Ho Chi Minh City (HCMC), Hanoi and Dong Thap were screened for antimicrobial residues by PremiTest, and were further investigated by Charm II. Samples from HCMC (N=113) were cultured using ISO 6579:2002/Amd 1:2007. NTS bacteria were quantified using a minimum probable number (MPN) technique. NTS isolates were assigned to serovar by Multilocus Sequence Typing (MLST), and were investigated for their phenotypic susceptibility against 32 antimicrobials. A total of 26 (7.3%) samples tested positive by PremiTest (9.5% beef, 4.1% pork and 8.4% chicken meat). Sulfonamides, tetracyclines and macrolides were detected by Charm in 3.1%, 2.8% and 2.0% samples, respectively. Overall, meat samples from wet markets had a higher prevalence of residues than those from supermarkets (9.6% vs. 2.6%) (p=0.016). NTS were isolated from 68.4% samples from HCMC. Chicken samples from wet markets had by far the highest NTS counts (median 3.2 logMPN/g). NTS isolates displayed high levels of resistance against quinolones (52.2%) and β-lactams (49.6%), but low levels against 3rd generation cephalosporins (4.4%) and aminoglycosides (0.8%). The highest adjusted prevalence of multidrug resistance (MDR) corresponded to isolates from chicken meat and pork (OR 8.3 and 1.8, respectively) (baseline=beef). S. Kentucky was the most common serovar identified (11 from chicken, 1 from beef) and 91.7% isolates was MDR. 11/12 isolates corresponded to ST198, a worldwide-disseminated multi-resistant NTS clone. We recommend stepping up policy measures to promote responsible antimicrobial use in animal production, as well as awareness about withdrawal periods to limit the hazard of residues in animal products, and improving slaughtering/hygiene procedures to limit cross-contamination with NTS, particularly in poultry wet markets.

The Possible Influence of Non-synonymous Point Mutations within the FimA Adhesin of Non-typhoidal Salmonella (NTS) Isolates in the Process of Host Adaptation.

Non-typhoidal Salmonella (NTS) remains a global pathogen that affects a wide range of animal species. We analyzed a large number of NTS isolates of different host origins, including Salmonella Heidelberg (n = 80, avian), S. Dublin (50, bovine), S. Typhimurium var 5- (n = 40, porcine), S. 4,5,12,:i:- (n = 40, porcine), S. Cerro (n = 16, bovine), and S. Montevideo (n = 14, bovine), using virulence profiling of the bcfC, mgtC, ssaC, invE, pefA, stn, sopB, and siiE virulence-associated genes, a biofilm production assay, pulsed field gel electrophoresis, and the full-length sequencing of the fimA (adhesin) and iroN (receptor) genes. We determined a key amino acid substitution, A169 (i.e., threonine changed to alanine at position 169), in the FimA protein that changed ligand affinity of FimA toward N-acetyl-D-glucosamine. This finding clearly indicates the important role of non-synonymous single nucleotide polymorphism (nsSNPs) in adhesin functionality that may impact the host tropism of NTS. This nsSNP was found in S. Heidelberg and S. Cerro isolates. Although this was not the case for the IroN receptor, the phylogeny of this receptor and different host origins of NTS isolates were positively correlated, suggesting existence of specific host immune selective pressures on this unique receptor in S. enterica. We found that pefA, a gene encoding major fimbrial subunit, was the most-segregative virulence factor. It was associated with S. Heidelberg, S. Typhimurium var 5- and S. 4,5,12,:i:- but not with the rest of NTS strains. Further, we observed a significantly higher frequency of non-biofilm producers among NTS strains that do not carry pefA (42.5%) compared to S. Heidelberg (2.5%) and S. Typhimurium var 5- (7.5%) and S. 4,5,12,:i:- (0%). This study provides new insights into the host adaptation of avian and mammalian NTS isolates that are based on the bacterial antigens FimA and IroN as well as the interrelationships between host adaptation, overall genetic relatedness, and virulence potential in these NTS isolates.

Molecular characterization of antimicrobial resistant non-typhoidal Salmonella from poultry industries in Korea

BackgroundAntimicrobial resistant Salmonella strains are a direct threat to human health when this resistance interferes with treatment and an indirect threat when resistance can be transferred to other human pathogens. The objective of the present study was to characterize antimicrobial resistant non-typhoidal Salmonella (NTS) isolates recovered from poultry industries, including a description of genetic diversity and virulence profiles.ResultsIn total of 93 Salmonella isolates shown antimicrobial resistance to one or more drugs, all isolates exhibited common resistance to streptomycin, nalidixic acid and cephalothin but no ciprofloxacin resistance. Among 26 virulence gene profiling, 12 virulence genes, invA, orgA, prgH, sopB, tolC, sipB, gatC, msgA, pagC, spiA, sifA, and sitC were found in all antimicrobial-resistant NTS isolates. In comparing the data from ERIC-PCR clusters, virulence profiles and resistance profiles, some Salmonella isolates grouped into the same cluster were found to exhibit similar virulence and resistance patterns.ConclusionsVirulence profiling combined with ERIC-PCR offered a rapid approach to characterize antimicrobial-resistant NTS.

Strong influence of livestock environments on the emergence and dissemination of distinct multidrug-resistant phenotypes among the population of non-typhoidal Salmonella.

The problem of emergence and dissemination of multidrug resistance, especially among Gram-negative bacteria, has reached alarming levels. This increases the need to develop surveillance methods that more effectively and accurately provide information about the emergence and spread of multidrug-resistant organisms. In this study, using a well-defined population of non-typhoidal Salmonella (NTS) isolates associated with avian, bovine and porcine hosts, we found that the livestock environments had a specific (P < 0.005) and profound (P < 0.005) effect on the evolution of multidrug-resistant phenotypes among population of NTS isolates. The MDR pattern containing penicillins, tetracyclines and macrolides and the evolving counterparts (i.e., penicillins, tetracyclines and macrolides + other antibiotic classes) were significantly (P < 0.005) associated with NTS isolates of porcine origin. Similarly, MDR patterns containing folate pathway inhibitors, macrolides and aminocyclitol or containing penicillins, cephalosporins, tetracyclines, phenicols and macrolides were significantly associated with avian (P < 0.005) and bovine (P < 0.005) NTS isolates, respectively. Furthermore, STRUCTURE, an evolutionary analysis, clearly showed that the host origin (i.e., livestock environment), and not the genetic background of different NTS serovars, was the most determinative factor for acquisition and spread of MDR phenotypes. In addition, we described a novel non-synonymous mutation, located outside of the QRDR at position 864 of GyrA, that was likely associated with fluoroquinolone resistance.

Molecular characterization of antimicrobial multi-drug resistance in non-typhoidal Salmonellae from chicken and clam in Mangalore, India.

Salmonella enterica has been documented as one of the leading causes of salmonellosis throughout the world and is most commonly associated with the consumption of contaminated food products. Thus, this research was aimed at studying the antimicrobial susceptibility pattern and detection of quinolone resistance in Salmonella spp isolated from food of animal origin. Thirty-six Salmonella isolates comprising 8 from poultry and 28 from seafood (clams) were identified, serotyped and characterized for their antimicrobial susceptibility against 10 different antibiotics. Plasmid DNA was isolated from all the isolates by alkaline lysis, quinolone resistant non-typhoidal S. Weltevreden were examined for mutation in the DNA gyrase coding gene. Among the 36 Salmonella isolates, 20 were S. weltevreden (8 from poultry and 12 from seafood) and 16 were S. Typhimurium (from seafood). All the isolates showed multiple resistance to nalidixic acid, tetracycline, co-trimoxazole and nitrofurantoin, but, interestingly, the isolates were 100% susceptible to ampicillin, chloramphenicol and gentamicin. Resistant isolates from the study carried the genes responsible for resistance to respective antibiotics. The strain S130 isolated in the study showed single point mutation, Asp87Gly, at position 87 in quinolone resistance determining region. It revealed mutation in quinolone resistance determining region as a cause for quinolone resistance in non-typhoidal Salmonellae. The occurrence of genes accountable for plasmid mediated resistance to quinolones (viz., qnrA, qnrB and qnrS) in plasmid of non-typhoidal Salmonellae isolates provides evidence for plasmid mediated quinolone resistance.

Emergence of mcr-1 among Nontyphoidal Salmonella Isolates in the United States

Background Colistin is considered a critically important antimicrobial for its role in the treatment of severe multidrug-resistant infections. Colistin resistance conferred by the plasmid-mediated gene mcr-1 has been reported in enteric pathogens globally since 2015, but remains rare in the United States. We describe the search for mcr-1 among nontyphoidal Salmonella (NTS) and the identification of the first human cases in the United States.Methods Whole genome sequencing (WGS) was performed on NTS isolates from humans by state health departments, from retail meat by the US Food and Drug Administration, and from food animals by the US Department of Agriculture. Sequences were uploaded to the National Center for Biotechnology Information and screened through their pathogen detection pipeline for the presence of resistance determinants (including mcr-1) beginning in late 2015; screening included some retrospective sequences. Isolates with the suspected mcr-1 gene were submitted to CDC for confirmatory PCR. Epidemiological information on human cases was collected from state health departments.ResultsOver 70,000 Salmonella isolates from humans, retail meat, and food animals were screened for mcr-1. No NTS with mcr-1 were identified in retail meat or food animals. Four human cases of NTS with mcr-1 were identified by WGS and three were confirmed by PCR (1 pending testing): Salmonella Corvallis in an 18-year-old man from Tennessee (isolation July 2014), Salmonella Enteritidis in a 55 year-old woman from Connecticut (isolation May 2016), Salmonella Typhimurium in a 57-year-old woman from Virginia (isolation November 2016), and Salmonella Enteritidis in a 47-year-old man from Minnesota (isolation April 2017). All patients traveled internationally in the 10 days prior to illness onset.Conclusion NTS rarely contain mcr-1 in the United States. To date, all human cases have been linked to international travel, reflecting the higher prevalence of mcr-1 reported from other parts of the world. The absence of mcr-1 in NTS from US food animals and retail meat is likely because colistin has not been used in food animal agriculture in the United States, underscoring the importance of a One Health approach to combat antimicrobial resistance.Disclosures All authors: No reported disclosures.

Molecular Detection of Mutations within the Quinolone Resistance-Determining Regions in Non Typhoidal Salmonella Isolates from Malaysia

Introduction: The efficacy of chemotherapy in bacteraemia caused by non-typhoidal Salmonella (NTS) is compromised by antibiotic resistance. Objective: This study was undertaken to describe the mechanism of resistance among clinical NTS isolates. Materials & Methodology: Thirty of NTS were isolated from blood (n = 19), stool (n = 10) and bronchioalveolar lavage (BAL; n = 1) respectively. These isolates were tested for susceptibility testing by disc diffusion method against ampicillin, gentamicin, tetracycline, co-trimoxazole, nalidixic acid, ciprofloxacin and ceftriaxone. Epsilometer tests (E-test) for nalidixic acid and ciprofloxacin were performed for nalidixic acid resistant isolates by disc diffusion method. DNA sequencing was carried out on six of the nalidixic acid resistant Salmonella Enteritidis isolates to identify mutations within quinolones resistance determining regions (QRDR) of gyrA, gyrB, parC and parE genes. Results: Resistance rates of NTS isolates from blood, stool, and BAL were respectively 37%, 20% and 0% for ampicillin, 79%, 40% and 0% for tetracycline, 32%, 40% and 0% for co-trimoxazole, 37%, 10% and 100% for nalidixic acid. Eight isolates were resistant to nalidixic acid and had exhibited reduced susceptibility towards ciprofloxacin by E-test. Mutation within QRDR was detected in gyrA gene (n = 6; Asp 47 → His [3], Asp 51 → Asn [1], Asp 73 → Gly [1], and Gly 48 → Asp [1]) and double mutation was detected in parE gene (n = 3; Gly 48 → Asp [3], Glu 82 → Ser [3]). Out of six isolates, three isolates were found to have both gyrA and parE gene mutations. Conclusions: There was no mutation observed in gyrB and parC gene. Mutation in gyrA gene was sufficient to induce decreased susceptibility to ciprofloxacin. Variation in amino acid sequences are novel, while detection of other gene mutation was uncommon.

Microbiological, clinical and molecular findings of non-typhoidal Salmonella bloodstream infections associated with malaria, Oriental Province, Democratic Republic of the Congo.

BackgroundIn sub-Saharan Africa, non-typhoidal Salmonella (NTS) can cause bloodstream infections, referred to as invasive non-typhoidal Salmonella disease (iNTS disease); it can occur in outbreaks and is often preceded by malaria. Data from Central Africa is limited.MethodsClinical, microbiological and molecular findings of NTS recovered in a blood culture surveillance project (2009–2014) were analyzed.ResultsIn March-July 2012 there was an epidemic increase in malaria infections in the Oriental Province of the Democratic Republic of the Congo (DRC). In one referral hospital, overall hospital admissions in June 2012 were 2.6 times higher as compared to the same period in the years before and after (336 versus an average of 128 respectively); numbers of malaria cases and blood transfusions were nearly three- and five-fold higher respectively (317 versus 112 and 250 versus 55). Case fatality rates (in-hospital deaths versus all admissions) peaked at 14.6 %. Salmonella Typhimurium and Salmonella Enteritidis together accounted for 88.9 % of pathogens isolated from blood cultures collected during an outreach visit to the affected districts in June 2012. Children infected with Salmonella Enteritidis (33 patient files available) tended to be co-infected with Plasmodium falciparum more often than children infected with Salmonella Typhimurium (40 patients files available) (81.8 % versus 62.5 %). Through the microbiological surveillance project (May 2009–May 2014) 113 unique NTS isolates were collected (28.5 % (113/396) of pathogens); most (95.3 %) were recovered from children < 15 years. Salmonella Typhimurium (n = 54) and Salmonella Enteritidis (n = 56) accounted for 47.8 % and of 49.6 % NTS isolates respectively. Multilocus variable-number tandem-repeat analysis (MLVA) revealed more heterogeneity for Salmonella Typhimurium than for Salmonella Enteritidis. Most (82/96, 85.4 %) NTS isolates that were available for antibiotic susceptibility testing were multidrug resistant. All isolates were susceptible to ceftriaxone and azithromycin.ConclusionDuring the peak of an epidemic increase in malaria in the DRC in 2012, a high proportion of multidrug resistant Salmonella Typhimurium and Salmonella Enteritidis were isolated from blood cultures. Overall, the two serovars showed subtle differences in clinical presentation and genetic diversity.Electronic supplementary materialThe online version of this article (doi:10.1186/s12879-016-1604-1) contains supplementary material, which is available to authorized users.

Physiological fluid shear alters the virulence potential of invasive multidrug-resistant non-typhoidal Salmonella Typhimurium D23580.

Salmonella enterica serovar Typhimurium strains belonging to sequence type ST313 are a major cause of fatal bacteremia among HIV-infected adults and children in sub-Saharan Africa. Unlike “classical” non-typhoidal Salmonella (NTS), gastroenteritis is often absent during ST313 infections and isolates are most commonly recovered from blood, rather than from stool. This is consistent with observations in animals, in which ST313 strains displayed lower levels of intestinal colonization and higher recovery from deeper tissues relative to classic NTS isolates. A better understanding of the key environmental factors regulating these systemic infections is urgently needed. Our previous studies using dynamic Rotating Wall Vessel (RWV) bioreactor technology demonstrated that physiological levels of fluid shear regulate virulence, gene expression, and stress response profiles of classic S. Typhimurium. Here we provide the first demonstration that fluid shear alters the virulence potential and pathogenesis-related stress responses of ST313 strain D23580 in a manner that differs from classic NTS.

Potential Sources and Transmission of Salmonella and Antimicrobial Resistance in Kampala, Uganda.

In sub‒Saharan Africa, non‒typhoidal Salmonellae (NTS) cause invasive disease particularly in children and HIV infected adults, but the disease epidemiology is poorly understood. Between 2012 and 2013, we investigated NTS sources and transmission in Kampala. We detected Salmonella in 60% of the influent and 60% of the effluent samples from a wastewater treatment plant and 53.3% of the influent and 10% of the effluent samples from waste stabilization ponds that serve the human population; 40.9% of flush‒water samples from ruminant slaughterhouses, 6.6% of the poultry fecal samples from live bird markets and 4% of the fecal samples from swine at slaughter; and in 54.2% of the water samples from a channel that drains storm–water and effluents from the city. We obtained 775 Salmonella isolates, identified 32 serovars, and determined resistance to 15 antimicrobials. We genotyped common serovars using multiple‒locus variable number tandem repeats analysis or pulsed‒field gel electrophoresis. In addition, we analyzed 49 archived NTS isolates from asymptomatic livestock and human clinical cases. Salmonella from ruminant and swine sources were mostly pan‒susceptible (95%) while poultry isolates were generally more resistant. Salmonella Kentucky isolated from poultry exhibited extensive drug resistance characterized by resistance to 10 antimicrobials. Interestingly, similar genotypes of S. Kentucky but with less antimicrobial resistance (AMR) were found in poultry, human and environmental sources. The observed AMR patterns could be attributed to host or management factors associated with production. Alternatively, S. Kentucky may be prone to acquiring AMR. The factors driving AMR remain poorly understood and should be elucidated. Overall, shared genotypes and AMR phenotypes were found in NTS from human, livestock and environmental sources, suggesting zoonotic and environmental transmissions most likely occur. Information from this study could be used to control NTS transmission.

Emergence and Diversity of Salmonella enterica Serovar Indiana Isolates with Concurrent Resistance to Ciprofloxacin and Cefotaxime from Patients and Food-Producing Animals in China.

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.

Association of Phenotypic and Genotypic Fluoroquinolone Susceptibility among Non-Typhoidal Salmonella Isolates from Jeddah, KSA

Non-typhoid Salmonella (NTS) infects 250 to 3200 per 100,000 individuals from all over the world. NTS infection is relatively high in Jeddah (ranges between 44-132/100,000 population) as compared to other cities of Saudi Arabia. NTS isolates have also shown increasing resistance to conventional antibiotics. Therefore, fluoroquinolone (FQ) is considered drug of choice for the treatment of invasive NTS infections. A rapid detection of FQ resistance may greatly assist in appropriate therapy and containment of resistant NTS strains. Thus, molecular detection of mutations in FQ resistance genes (gyrA and parC) may play a promising role. Since limited data were available about FQ resistance among NTS isolates, therefore, this study primarily explored the occurrence of phenotypic and genotypic FQ resistance among NTS isolates from Jeddah, Saudi Arabia. Study also explored any correlation between phenotypic and genotypic FQ resistance. Fifty NTS isolates were collected from a public sector hospital of Jeddah from January to December, 2014. FQ susceptibility was determined for 48 NTS isolates using Kirby-bauer disk diffusion method and results were interpreted according to Clinical and Laboratory Standards Institute (CLSI) methodology. Genotypic resistance to FQ was determined by exploring mutations in gyrA and parC genes using PCR- based gene-sequencing method. Results were statistically analyzed using Social Package of Statistical Science (SPSS) version 22. Phenotypic antibiogram revealed that 38% (18/48) NTS isolates were FQ resistant, that 23% (11/48) were intermediately susceptible and that 39% (19/48) were susceptible. Genotypic resistance revealed mutations in only four codons of gyrA and parC genes among 39% (7/18) of FQ resistant isolates. 43% (3/7) of FQ resistant isolates showed mutations at two codons 83 (S83F, S83Y) and 87, (D87G, D87Y, D87W) of gyrA gene. Two resistant isolates showed triple mutations i.e. at codons 83 and 87 of gyrA and codon 80 (S80I and S80W) of parC gene, while one resistant isolate revealed mutation at codon 87 of gyrA and 57 (S57T) of parC gene. Moreover, 55% (6/11) intermediately susceptible isolates for FQ also revealed mutation at codon 83 of gyrA gene whereas; one intermediately susceptible isolate (1/6) also revealed additional mutation at codon 57 of parC gene. None of the FQ susceptible NTS isolates showed any mutations in gyrA or parC genes. Occurrence of mutations at only four codons in gyrA and parC genes among FQ resistant isolates may assist in development of rapid molecular method for FQ resistance detection. Presence of mutations among more than fifty percent of intermediately susceptible FQ isolates is alarming and may serve as a predictor for pre-resistant isolates for FQ. Moreover, absence of mutation in about sixty percent of phenotypically FQ resistant NTS isolates shows existence of an alternate resistance mechanism requiring further investigations.