Molecular Screening of Carbapenem-Resistant K. pneumoniae (CRKP) Clinical Isolates for Concomitant Occurrence of Beta-Lactam Genes (CTX-M, TEM, and SHV) in the Kingdom of Bahrain.

The emergence of extended-spectrum ß-lactamase-producing Klebsiella pneumoniae, including CRKP infections, has resulted in significant morbidity and mortality worldwide. We aimed to explore the presence of bla genes (CTX-M, TEM, and SHV) in CRKP isolates. A total of 24 CRKP isolates were randomly selected from the Salmaniya Medical Complex Microbiology Laboratory. These isolates, which were positive for carbapenemases, were further explored for CTX-M, TEM, and SHV genes using PCR. All the CTX-M PCR amplicons were sent for sequencing. To determine genetic relatedness, molecular typing by ERIC-PCR was performed. The bla gene testing demonstrated that a significant proportion of these isolates harbored SHV, CTX-M, and TEM genes (100%, 91.6%, and 45.8%), respectively. Bioinformatic analyses confirmed CTX-M-15 in these isolates. ERIC-PCR analysis showed three clusters demonstrating genetic relatedness. The study findings reveal the concomitant carriage of the SHV and CTX-M-15 and a comparatively lower carriage of TEM genes in CRKP isolates. Our findings highlight the significance of routinely reporting the presence of antibiotic resistance genes along with regular antibiotic sensitivity reports, as this will aid clinicians in prescribing appropriate antibiotics.

Pattern of upper respiratory tract infections and physicians' antibiotic prescribing practices in Bahrain.

OBJECTIVE: To evaluate the pattern of antibiotic prescriptions for paediatric upper respiratory tract infections (URTI) and determine the associated predictors for such antibiotic use in the Kingdom of Bahrain. SUBJECTS AND METHODS: From March 2005 to March 2006, demographic data, clinical presentation, investigations and antibiotic prescription for children with URTI (n = 184) at the Bahrain Defence Force Hospital was recorded. To assess the factors which influence physician antibiotic prescription for URTI, a cross-sectional survey of doctors was carried out using a pre-tested questionnaire which was administered to paediatricians, general practitioners and emergency room physicians. RESULTS: Antibiotics were given to 95 of the 184 (51.6%) patients, mainly children <3 years (40/95). Significant association was demonstrated for antibiotic prescription, age and diagnosis of tonsillitis or acute otitis media (p < 0.05). Amoxicillin (37/95) was the most frequently prescribed antibiotic, followed by beta-lactam/beta-lactamase combination and second-generation cephalosporins. Fever, younger age, sore throat and presence of earache increased the likelihood of antibiotic prescription. Data from the cross-sectional survey of doctors revealed that lack of national guidelines, parental pressure and diagnostic uncertainty contributed to antibiotic overuse. CONCLUSION: Antibiotic overuse for the treatment of paediatric URTI remains a problem in our setting. We suggest the development of national guidelines which are integrated with structured continuing medical education courses, public awareness campaigns and the introduction of rapid streptococcal antigen tests in the outpatient clinic.

Antimicrobial prescribing trends in primary care: implications for health policy in Bahrain.

PURPOSE: To evaluate antimicrobial prescribing pattern by primary care physicians. METHODS: A nation-wide, retrospective, multi-centric prescription-audit was carried out in primary care health centres in Bahrain. RESULTS: Systemic antimicrobials ranked the fourth most common class of drugs prescribed. Amoxycillin, cephalexin, erythromycin, ciprofloxacin and cotrimoxazole were prescribed by general practitioners (GPs) more often than by family physicians (FPs) (p < 0.05). With respect to prescribing of other antimicrobials and anthelmintic mebendazole, the differences between GPs and FPs were nonsignificant. Seventy-seven per cent of systemic antimicrobials prescribed were for respiratory tract infections (RTIs). Topical antimicrobial preparations for ear and eye infections were prescribed by GPs in a rate significantly higher than by FPs (p < 0.05); of these, chloramphenicol and Locacorten vioform (flumethasone + clioquinol) ear drops and sulphacetamide eye drops were more often prescribed by GPs (p < 0.05). There were no significant differences in prescribing between GPs and FPs as regards topical antimicrobials used for oropharyngeal, skin and vulvovaginal infections. CONCLUSION: Antimicrobials were extensively used in primary care, mainly for treating RTIs. The general practitioners were more avid prescribers of antimicrobials compared to the FPs. Rational use of antimicrobials in primary care should be encouraged and the reasons for the observed differences in prescribing of antimicrobials between the GPs and FPs need further evaluation.

Management of acute diarrhoea in primary care in Bahrain: self-reported practices of doctors.

This nationwide study was conducted to assess the extent of adherence of primary-care physicians to the World Health Organization (WHO)-recommended guidelines on the use of oral rehydration therapy (ORT), antimicrobials, and prescribing of other drugs used in treating symptoms of acute diarrhoea in Bahrain. A questionnaire-based, cross-sectional survey was carried out in primary-care health centres. During a six-week survey period (15 August-30 September 2003), 328 (25.2%) completed questionnaires were returned from 17 of 20 health centres. In a sample of 300 patients, oral rehydration salts (ORS) solution was prescribed to 89.3% (n=268) patients; 12.3% received ORS alone, whereas 77% received ORS in combination with symptomatic drugs. Antimicrobials were prescribed to 2% of the patients. In 11.4% of the cases, rehydration fluids and other drugs were given parenterally The mean number of drugs was 2.2+0.87 per prescription. In approximately one-third of the patients, three or more drugs were used. Primary-care physicians almost always adhered to the WHO guidelines with respect to ORT and antimicrobials. However, in several instances, ORT was prescribed along with polypharmacy, including irrational use of drugs for symptomatic relief. Effective health policies are needed to reduce the unnecessary burden on the healthcare system.

Invasive capabilities of Campylobacter jejuni strains isolated in Bahrain: molecular and phenotypic characterization.

The association between putative virulence genes in Campylobacter jejuni clinical isolates, in vitro invasive capability and severity of infection is yet to be clearly described. We have characterized three virulence genes and correlated their presence with the severity of infection and in vitro invasiveness. We studied eight C. jejuni strains isolated from patients whose clinical data were scored to determine severity of infection. Cytolethal distending toxin (cdtB), invasion associated marker (iam) and Campylobacter invasion antigen (ciaB) genes were detected by PCR and INT407 cells used for invasion assays. Two strains positive for all three genes were the most invasive and isolated from patients with the most severe infection. Four strains positive for two genes and two strains negative for all the three genes were identified. The two cdtB(+ve)/ciaB(+ve) strains were more invasive than the cdtB(+ve)/iam(+ve) strains. One of the cdtB(-ve)/ciaB(-ve) strains showed invasion levels similar to cdtB(+ve)/ciaB(+ve) strains, but the second strain had a non-invasive phenotype. The findings indicate a correlation between in vitro invasive capability, and the presence of all three genes. The pattern of association between invasiveness and molecular characterization suggests that the ciaB gene confers a more invasive capability.

Clinical relevance of virulence genes in Campylobacter jejuni isolates in Bahrain.

There are no data describing the genetic make-up of Campylobacter strains (an important aetiological agent of diarrhoea) circulating in the Arabian Gulf region. Here, the molecular characterization of two virulence genes in Campylobacter jejuni from Bahrain and the relationship with clinical infection are reported. Molecular screening for cytolethal distending toxin (cdtB) and invasion-associated marker (iam) genes was carried out on C. jejuni stool isolates collected from January 2002 to January 2004 in Bahrain. The molecular characterization was correlated with the patients' socio-demographic and clinical parameters. Of the 96 C. jejuni strains tested, 50 (52 %) were cdtB+/iam+, 30 (31 %) were cdtB+/iam- and 16 (17 %) were cdtB-/iam-. Sixty-nine per cent (66/96) of patients were less than 3 years old, with significantly higher detection of cdtB+/iam+ and cdtB+/iam- strains (P < 0.001 and P < 0.01, respectively) in this age group. Seventy patients (73 %) were symptomatic. In the group that were less than 3 years old, 62 and 85 % of those with cdtB+/iam+ and cdtB+/iam- strains, respectively, were symptomatic compared with 100 % for those over 3 years of age. However, the presence of cdtB-/iam- strains still resulted in clinical infection in the children under 3 years but not in the older patients. This is the first report describing the molecular characterization of virulence genes in Campylobacter isolates from this region. The findings indicate that strains of different virulence genetic make-up are circulating in the population, with children under the age of 3 years being most vulnerable. Further work on the molecular characterization, gene expression and determination of the invasive phenotypes of C. jejuni strains circulating in different regions is needed.

Effect of antibiotic sub inhibitory concentration on cytolethal distending toxin production by Campylobacter jejuni.

OBJECTIVES: To investigate the effect of pre-exposure antibiotics on cytolethal distending toxin (CDT) production and toxigenic effect of C. jejuni. METHODS: Sonicates and filtrates were prepared from known cdt+ and cdt- isolates of C. jejuni which had been pre-exposed to varying concentrations (MIC, 1/2 MIC, 1/4 MIC, 1/8 MIC) of erythromycin and ciprofloxacin. The CDT toxigenic effect was examined using INT 407 and HeLa cells. RESULTS: A trend of increased toxigenic effect was observed with pre-exposure to antibiotics. This was more pronounced with erythromycin pre-exposure compared to ciprofloxacin. Although a trend of increasing toxigenic effect with decreasing antibiotic concentration was demonstrable, some differences were observed between isolates. In one isolate the increased toxigenic effect was statistically significant (P<0.05) at 1/4 MIC in INT 407 cells and at 1/8 MIC in HeLa cells. CONCLUSIONS: This study provides evidence of an association between CDT production by C. jejuni and pre-exposure to antibiotics. Pre-exposure to ciprofloxacin and erythromycin at concentrations below MICs could potentiate CDT activity. Further work is needed to elucidate the mechanism involved. We recommend that these antibiotics be used in the treatment of C. jejuni enteritis only when strongly indicated and with careful monitoring of patients.

Status of malaria in the Kingdom of Bahrain: a 10-year review.

BACKGROUND: Malaria was eradicated in Bahrain about 20 years ago. However, because of a large immigrant population, cases of imported malaria continued to be seen. This paper presents an evaluation of the data from 1992 to 2001 to assess the levels of indigenous and imported malaria cases and the potential for reemergence of malaria transmission in the country. METHODS: Epidemiologic and parasitologic data on confirmed malaria cases during the review period were analyzed. Data on vector breeding activity were also analyzed to determine the potential for reemergence of local transmission. RESULTS: From 1992 to 2001, 1,572 cases of malaria were reported. All were imported malaria cases. There was a consistent decline in the number of cases, from 282 (the peak level) in 1992 to 54 in 2001. Eighty-four percent (1,318/1,572) of infections were contracted as a result of travel to or previous domicile in five countries, namely India, Pakistan, Sri Lanka, Bangladesh and Sudan. Plasmodium vivax was the etiologic agent in 85.6% (1,346/1,572) of cases, with Plasmodium falciparum accounting for 14% (220/1,572). During this period, the percentage of examined breeding sites where Anopheles mosquito larvae were identified remained fairly constant, ranging between 0.05% and 0.1%. CONCLUSIONS: From 1992 to 2001, there was a consistent decline in the number of imported malaria cases in Bahrain. There were no cases of locally acquired malaria during this period. The low level of vector breeding spots with Anopheles mosquito larvae suggests that the potential for reemergence of local transmission remains low.

Evaluation of malaria chemoprophylaxis in Bahrain.

The Kingdom of Bahrain is an archipelago located in the Arabian Gulf with a population of 650,000. A well-established network of 21 primary health care centers provide free health care for all residents, including citizens and expatriates. Patients requiring special investigation, consultation with specialists or admission are referred to the Salmaniya Medical Complex (SMC), which provides secondary and tertiary care. Malaria is a medical condition referred to SMC for medical care. Advice on appropriate prophylactic antimalarial regimens is a service provided by and limited to a public health advisory body, including the Public Health Directorate in the Ministry of Health, and a mini-clinic known as a vaccination clinic located at one of the 21 primary health centers spread across the country. This clinic is staffed by a trained nurse; it provides services such as vaccination and malaria chemoprophylaxis advice. The recommended prophylactic drugs have to be purchased by the travelers from private pharmacies; they are considered to be over-the-counter (OTC) medications.

Causative pathogens of severe diarrhea in children.

OBJECTIVES: To investigate the enteropathogens in children with diarrhea attending Salmaniya Medical Complex, Bahrain. METHODS: Fecal samples from 805 children up to 15 years were examined for parasites, ova and cysts by direct wet preparation, formol-ether concentration and modified Ziehl-Neelsen stain, during the period November 1998 through to June 2000. Samples were cultured for Salmonella, Shigella, Campylobacter and Enteropathogenic Escherichia coli. Antibiotic sensitivity tests were performed on the relevant clinical isolates by agar disk diffusion method. All stools from children below 3 years of age (653 samples) were processed for adenovirus and rotavirus using a commercially available latex agglutination test (Diarlex). In addition, reverse transcription-polymerase chain reaction was performed on 200 randomly selected samples using oligonucleotide primers for Rotavirus A, B and C. RESULTS: Four subjects were found positive for parasites. Eighty-three (10.3%) samples were found positive for Salmonella (46 isolates), Shigella (26 isolates), Campylobacter jejuni (7 isolates), and Enteropathogenic Escherichia coli (4 isolates). Rotavirus was found in 91 (13.9%) samples and 4 samples (0.6%) were found positive for adenovirus. Out of 200 samples examined by reverse transcription-polymerase chain reaction, 73 (36.5%) were positive for group A rotavirus. CONCLUSIONS: Rotavirus type A appeared to be the most common single agent in our pediatric population, followed by the classical bacterial pathogens. Adenovirus and parasites appeared to play a very minor role in diarrhea. Thus, we suggest the introduction of rotavirus diagnostic tests in microbiological examination of diarrheic stools of children below 3 years of age.