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Aims@#OpenPCR is a low cost yet accurate thermocycler which can be self-built. The aim of the study is to highlight a low-cost alternative method for rapid confirmation of five predominant non-typhoidal Salmonella (NTS) serotypes using a multiplex PCR on a portable-DIY OpenPCR© thermocycler. @*Methodology and results@# Eight multiplex polymerase chain reaction (mPCR) samples containing genomic DNA of S. Agona selectively placed on the wells of the conventional PCR and OpenPCR© thermocyclers showed uniform heating in both thermocyclers. The limit of detection was similar for both thermocyclers for all five serotypes. The limit of detection for S. Typhimurium, S. Agona and S. Weltevreden was 10 pg/µL whereas the limit of detection for S. Enteriditis and S. Heidelberg was 1 pg/µL and 100 pg/µL, respectively. This assay incorporated a panel of unique genes; STM4495, SEN1392, SeHa-C4893, SeAg-B1096 and SENTW-3241 which were previously identified to be specific for S. Typhimurium, S. Enteritidis, S. Heidelberg, S. Agona, and S. Weltevreden, respectively, as well as the pan-Salmonella gene invA as internal control (IC) and pan-bacteria gene 16S rRNA to serve as amplification control (AC). The analytical specificity of the mPCR assay was found to be 100% for all five NTS using OpenPCR© thermocyclers. @*Conclusion, significance and impact of study@#The feasibility and low cost of the OpenPCR© thermocycler makes this device an ideal alternative for mPCR assay for rapid confirmation of NTS serotypes.
Subject(s)
Salmonella InfectionsABSTRACT
Resumen La meningitis por Salmonella spp. es infrecuente en pediatría, pero debe sospecharse especialmente en menores de seis meses. Salmonella subespecie enterica es la más frecuente a nivel mundial, siendo el serovar Enteritidis el más importante. Tiene una alta tasa de complicaciones, predominando las colecciones y abscesos, por lo que se recomienda realizar una neuroimagen a todos los pacientes. A pesar que el manejo no está estandarizado, hay consenso en que el tratamiento de elección son las cefalosporinas de tercera generación. La resistencia antimicrobiana es infrecuente pero va en aumento, principalmente en países subdesarrollados. Presentamos el primer caso publicado en Chile de una meningitis por S. Enteritidis en un lactante de 11 meses, que se presentó con fiebre y una convulsión focalizada, con hemocultivos y cultivo de líquido cefalorraquídeo positivos. El paciente completó tratamiento con ceftriaxona y ciprofloxacina por cuatro semanas con buena respuesta.
Abstract Salmonella spp meningitis is rare in pediatrics. However, it should be especially suspected in children younger than 6 months. The enteric subspecies is the most frequent worldwide, with the Enteritidis serovar being the most important. The complication rate is high, with collections and abscesses predominating. For this reason, neuroimaging is currently recommended for all patients. Even though management is not standardized, there is consensus that first line treatment should be with third generation cephalosporins. Antibiotic resistance is infrequent but increasing, mainly in underdeveloped countries. We present the first case reported in Chile of meningitis by Salmonella Enteritidis in an 11-month-old infant presenting with fever and focused epileptic crisis. Blood culture and cerebrospinal fluid culture were positive. The patient completed four weeks of treatment with ceftriaxone and ciprofloxacin with a good response.
Subject(s)
Humans , Infant , Salmonella Infections/diagnosis , Salmonella Infections/drug therapy , Meningitis, Bacterial/diagnosis , Meningitis, Bacterial/drug therapy , Salmonella enteritidis , Chile , Anti-Bacterial Agents/therapeutic useABSTRACT
Purpose:Infections due to invasive non-typhoid salmonella can be dangerous and fatal. The mode of infection and the severity varies from the typhoidal fevers. It is important to find the association between clinical features and the infecting serovar to understand the pathophysiology and course of treatment Methods:In the present study, extra-intestinal specimens (blood, cerebrospinal fluid and pus) from three patients suffering from septicaemia, meningitis and osteomyelitis were received. Micro-biological and biochemical test for species identification and antibiotic susceptibility was done as per standard protocol.Further, PCR based amplification and sequencing of a portion of the flagellin gene (FliC) was done to confirm the serovar.Results: Salmonellaentericawas identified from all the threeby microbiological and biochemical examination.The sequence of the Flic gene confirmed the serovar to be S.typhimurium. All the patients were treated successfully for the infectionby appropriate antibiotic therapy. Conclusion:The study highlights that serovarTyphimurium is common in invasive non-typhoidal salmonellosis and its pathophysiology and virulence factors expression should be understood in various organ types for better treatment options and outcomes
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Background: Non-typhoidal Salmonella (NTS) infection is a serious public health problem globally. Although NTS infections are self-limited, antimicrobial therapy is recommended for severe infections and immunocompromised patients. Antimicrobial resistance (AMR) in these pathogens further limits its therapeutic options. Here, we report an incidence of ceftriaxone resistance in NTS over the past 9 years in a southern Indian region. Materials and Methods: Molecular mechanisms of resistance in ceftriaxone-resistant NTS have been tested by both phenotypic and molecular methods. Minimum inhibitory concentration was determined by the E-test and broth microdilution method. AMR gene markers of ?-lactamases such as AmpCs (blaMOX, blaCMY, blaDHA, blaFOX, blaACC and blaACT) and extended-spectrum ?-lactamases (ESBLs) (blaSHV, blaTEM, blaVEB, blaPER, blaCTXM-1like,blaCTXM-2like, blaCTXM-8like, blaCTXM-9like and blaCTXM-25like) were screened. The presence of IncH12 and IncI1 plasmid was also analysed. Results: The study reports a 5% prevalence of ceftriaxone resistance in NTS. The most common serogroup was Salmonella Group B followed by Salmonella Group E and Salmonella group C1/C2. The occurrence of blaCTX-M-1, blaTEM, blaCMY and blaSHV genes was observed in 54%, 54%, 48% and 3% of the isolates, respectively. Interestingly, few isolates carried dual resistance genes (ESBLs and AmpCs). IncH12 and IncI1 plasmid was identified in isolates carrying ESBL and AmpC genes, respectively. Conclusion: This study shows that ceftriaxone resistance is mainly mediated by ?-lactamases such as ESBL and AmpC. As the incidence of ceftriaxone resistance is rising gradually over the years, it is imperative to monitor the AMR in this species.
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Background: Salmonella enterica serovar Weltevreden has been a rare cause of acute gastroenteritis occurring worldwide. Here, we report an outbreak of food poisoning in a tea garden. Objectives: To determine the aetiological agent and risk factors responsible for the outbreak and to take necessary steps for prevention of future outbreaks. Materials and Methods: Affected area was visited by a team of microbiologists for collecting stool samples/rectal swabs from affected patients. Samples were processed by culture followed by confirmation of the isolates biochemically, automated bacterial identification system, conventional serotyping and molecular typing. Water samples were also processed for detection of faecal contamination. Antimicrobial susceptibility testing was performed by Kirby–Bauer disc diffusion technique according to the Clinical Laboratory Standard Institute guidelines. Results: The isolates were confirmed as S. enterica subspecies enterica serovar Weltevreden. They were found sensitive to ampicillin, amoxycillin‑clavulanic acid, ciprofloxacin, ofloxacin, norfloxacin, cefotaxime, ceftriaxone, co‑trimoxazole and doxycycline. Water samples showed high‑level faecal contamination. Source of outbreak was found to be drinking water contaminated with dead livestock. House to house visit was made for early diagnosis and treatment of the cases, awareness campaigning and chlorination of drinking water. Conclusions: This report emphasises the geographical distribution of this organism in Assam. As S. Weltevreden is widely distributed in domestic animals, people should be made aware of immediate reporting of any unusual death among the livestock and their safe disposal which can significantly reduce the incidence of non‑typhoidal salmonellosis in the country.
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Objective To investigate the antimicrobial resistance pattern of non-typhoidal Salmonella isolated from diarrhea cases in Guangdong province,China.The multidrug-resistant strains were analyzed by pulsed field gel electrophoresis(PFGE) typing.Methods All the non-typhoidal Salmonella strains isolated between 2009 and 2011 were serotyped,then the antimicrobial resistance was detected by the disk diffusion method and molecular typed by PFGE.Results 91.76％ (256/279) S.typhimurium isolates were multiple resistant to 3 and more antimicrobials.Forty S.typhimurium isolates were multiple resistant to 9 and more antimicrobials and 3 out of which were multiple resistant to all the 12 antimicrobials in vitro.96.91％ (94/97) Salmonella I4,5,12:i:-isolates were multiple resistant to 3 and more antimicrobials.Nine Salmonella I4,5,12:i:- isolates were multiple resistant to 9 and more antimicrobials and I out of which was multiple resistant to all the 12 antimicrobials1 in vitro.47％ (47/100) S.enteritidis isolates were multiple resistant to 3 and more antimicrobials.Only 1 S.enteritidis isolates was multiple resistant to 9 and more antimicrobials.4.27％ (27/632) non-typhoidal Salmonella isolates was resistant to ciprofloxacin,including 17 S.typhimurium and 6 Salmonella 14,5,12:i:- isolates.Also,there were 3 1.96％ ( 202/632 ) non-typhoidal Salmonella isolates was intermediary to ciprofloxacin.The PFGE patterns of the predominant strains which were highly resistant and multidrug-resistant had different genotypes and demonstrated significant genetic diversity.Conclusion The situation about the multiple antimicrobial resistances of non-typhoidal Salmonella in Guangdong province has showed the prevalent problem.The PFGE types of the multiple drug-resistant strains prompted these strains were come from different clones.This requires that we continue to strengthen the resistance monitoring and control of the rational use of antibiotics.
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BACKGROUND: The emergence of non-typhoidal Salmonella (NTS) with decreased susceptibilities to fluoroquinolone, ampicillin, or ceftriaxone has been reported worldwide. However, current surveillance studies of resistance among NTS in Korea are limited. Thus, the antimicrobial susceptibilities; resistance mechanisms such as extended-spectrum beta-lactamase (ESBL), plasmid-mediated AmpC beta-lactamase (PABL), and plasmid-mediated quinolone resistance (PMQR); and molecular epidemiologic characteristics were investigated in the present study. METHODS: National Institute of Health and National Veterinary Research and Quarantine Service collected NTS strains from 219 clinical and 293 non-clinical specimens from 2006 to 2008. The antimicrobial susceptibilities were determined using the Clinical and Laboratory Standards Institute disk diffusion test. ESBL, PABL, and qnr genotyping were performed using PCR and nucleotide sequencing. Pulsed-field gel electrophoresis was used for the molecular epidemiologic study. RESULTS: The resistance to ampicillin in clinical and non-clinical NTS was 49% and 18 to 47%, respectively. The resistance rates to trimethoprim-sulfamethoxazole in clinical and non-clinical NTS were 8% and 0 to 41%, respectively. The rates to extended-spectrum cephalosporin were 0 to 1%. One CTX-M-15-producing isolate and four CMY-2-producing isolates were detected. Notably, PFGE analysis showed four isolates carrying bla CMY-2, including one non-clinical strain had high clonality. Although the rate of ciprofloxacin resistance was very low, two qnrS1-carrying NTS strains were detected in non-clinical specimens. CONCLUSION: The resistance rates to ampicillin in both clinical and non-clinical NTS were high, while those to trimethoprim-sulfamethoxazole varied depending on the specimen. NTS strains harboring CTX-M-15-type ESBL or CMY-2-type PABL were detected even though the resistance rates to cephalosporins were very low. Four NTS strains carrying the blaCMY-2-gene implied zoonotic infection. Continuous effort to minimize transfer of resistance genes in NTS is necessary.
Subject(s)
Animals , Humans , Ampicillin , Bacterial Proteins , beta-Lactamases , Ceftriaxone , Cephalosporins , Ciprofloxacin , Diffusion , Electrophoresis, Gel, Pulsed-Field , Korea , Lifting , Polymerase Chain Reaction , Quarantine , Salmonella , Sprains and Strains , Trimethoprim, Sulfamethoxazole Drug CombinationABSTRACT
Non-typhoidal salmonella (NTS) is a common pathogen involved in food poisoning. It has various extraintestinal complications including encephalopathy. However, NTS associated encephalopathy is rarely reported. A previously healthy 14-year-old boy was admitted for altered level of consciousness. He had profuse watery diarrhea with fever on the day of admission. Physical examination was unremarkable except slightly increased bowel sounds and altered mental state. Cerebrospinal fluid analysis revealed mild pleocytosis. Meningoencephalitis was the presumptive diagnosis. His consciousness was recovered over 24 hours, bloody and mucoid diarrhea developed the next day after admission. NTS organism was isolated from his stool. After 1 week, he fully recovered neurologically but a mild watery diarrhea persisted at the time of discharge.
Subject(s)
Adolescent , Humans , Consciousness , Consciousness Disorders , Diarrhea , Dysentery , Fever , Foodborne Diseases , Leukocytosis , Meningoencephalitis , Physical Examination , Salmonella , Salmonella InfectionsABSTRACT
BACKGROUND: Non-typhoidal Salmonella (NTS) are important commensal microorganisms. We intended to investigate the prevalence and mechanisms of nalidixic acid resistance among NTS isolated from human and poultry/livestock. METHODS: A total of 151 Salmonella isolates (36 human and 115 livestock isolates, respectively) was tested for the Minimum inhibitory concentrations (MICs) of nalidixic acid, together with serotyping. As for the nalidixic acid resistant isolates, further studies were taken: MICs of ciprofloxacin, mutation analysis of gyrA and parC genes, and organic solvent tolerance test. RESULTS: Eighty-four isolates of 151 human and livestock isolates were resistant to nalidixic acid. The prevalence of nalidixic acid resistance and was 13.9% (5 of 36 isolates) in human isolates and 68.7% (79 of 151 isolates), in the livestock isolates respectively. Among 84 nalidixic acid-resistant isolates, the The prevalence of ciprofloxacin resistance in livestock isolates was 24.1% (1 resistant and 18 intermediate of 79 strains), but no ciprofloxacin resistance was found in 5 human isolates. Among 65 nalidixic acid resistant, ciprofloxacin-susceptible isolates, 3 (60%, of 5 human isolates) and 60 (100%, all livestock isolates) showed low level fluoroquinolone resistance (ciprofloxacin MIC, 0.125-1.0 microgram/microliter). Six types of point mutations were found in the analysis of DNA sequencing of the gyrA gene in the 84 isolates; 75 isolates showed point mutations on amino acid Ser 83 and/or Asp 87. On the other hand, no point mutation was found from the parC genes. Forty-seven nalidixic acid resistant isolates showed tolerance to organic solvents. CONCLUSIONS: Nalidixic acid resistance was a good marker of low level fluoroquinolone resistance. As for the severe NTS infection, MIC test for nalidixic acid would be required.
Subject(s)
Humans , Ciprofloxacin , Hand , Livestock , Microbial Sensitivity Tests , Nalidixic Acid , Point Mutation , Prevalence , Salmonella , Sequence Analysis, DNA , Serotyping , Solvents , ViperidaeABSTRACT
Objective To understand non-typhoid Salmonella in diarrhea patients from Guangdong province in order to timely discover the outbreaks caused by them as well as to grasp the serotypes, antibiotic resistance and pulse-field gel electrophoresis (PFGE) types of those strains isolated from this surveillance program. Methods Salmonella strains from diarrhea patients were detected and all the positive strains were tested by serum agglutination, antibiotic susceptibility and PFGE. Results 71 nontyphoid Salmonella strains were isolated from 1128 stoop samples, with a positive rate of 6.29 %. All the strains were divided into 29 serotypes, with Salmonella serotype enteritidis and typhimurium showing the most common serotypes. Most of the strains were sensitive to cephalosporins and quinolones. The antibiotic resistance rates of S. typhimurium were higher than S. enteritidis and S. stanley. Other than S. enteritidis, all the serotype strains did not have the same type of PFGE. 17 S. enteritidis strains digested by Xba Ⅰ were divided to 8 PFGE types while the PFGE 4 type appeared the most common one. 12 S. enteritidis strains were typed again by Sfi Ⅰ and Not Ⅰ , and there were still 3 groups of strains showing the same PFGE pattern. Conclusion Most of the infection caused by non-typhoidal Salmonella was sporadic in Guangdong province in 2007. Cephalosporins and quinolones seemed the best in curing the infection of non-typhoidal Salmonella at the clinics.
ABSTRACT
Clinical manifestations of Salmonella infection are variable such as enterocolitis, bacteremia, enteric fever, focal infection and asymptomatic carrier status. Among these presentations, enterocolitis is the most common clinical manifestation and can be diagnosed by the use of fecal specimens. Patients with severe infectious colitis are at increased risk of developing a colon perforation after colonoscopy due to colon trauma. We report a case of a 31-year-old woman with transverse colon perforation after colonoscopy that was diagnosed with enterocolitis caused by infection with non-typhoidal group D Salmonella.
Subject(s)
Adult , Female , Humans , Bacteremia , Colitis , Colon , Colon, Transverse , Colonoscopy , Enterocolitis , Focal Infection , Megacolon, Toxic , Salmonella , Salmonella Infections , Typhoid FeverABSTRACT
PURPOSE: As the incidence of non-typhoidal salmonella strains resistant to antibiotics has been increased, we attempted to investigate clinical aspects of non-typhoidal salmonella gastroenteritis and antibiotics resistance. METHODS: From January 2000 to June 2002, 99 children with positive stool culture of non-typhoidal salmonella were studied about clinical features, the incidence of antibiotics and multi-drug resistance and the difference of incidence of antibiotics resistance according to immune status. RESULTS: There were 66 males and 33 females. The majority of them were under 5 years of age (71%). 25 children were immunocompromised due to chemotherapy, steroid or immunosuppressive treatment. Serogroup D was the most common isolates (65%) followed by B (16%), C (8%) and E (8%). Resistance rate of 30% to ampicillin, 12% to chloramphenicol, 20% to trimethoprim- sulfamethoxazole (TMP-SMX), 11% to cefotaxime and 8% to cefixime were obtained. All isolates were susceptible to ciprofloxacine. Resistance rate to cefotaxime and cefixime in immunocompromised patients was 24% and 14.3% respectively, which were significantly higher compared to that in immunocompetent patients (6.8%, 5.6%, p<0.05). 11 isolates were resistant to three or more antibiotics. The incidence of multi-drug resistant isolates was significantly higher in immunocompromised patients (24%) than that of immunocompetent patients (6.8%). CONCLUSION: Because of the high prevalence of non-typhoidal salmonella strains resistant to ampicillin, chloramphenicol and TMP-SMX, third-generation cephalosporin might be the treatment of choice in non-typhoidal salmonella gastroenteritis. In particular, antibiotics should be carefully selected in immunocompromised patients because non-typhoidal salmonellas from them showed the higher incidence of antibiotic resistance and multi-drug resistance.