Multidrug-resistant Enterobacteriaceae colonising the gut of adult rural population in South India

Background: Multidrug-resistant (MDR) colonisers act as a reservoir for transmission of antibiotic resistance and are a source of infection. Exposure to antibiotics by the commensal flora renders them resistant. Antibiotic consumption and hospitalisation are two major factors influencing this. We studied, antibiotic-resistant bacteria colonising rural adult population who had restricted access to health care and presumably had low consumption of antibiotics. Aim: Detection of multidrug resistance genes of extended spectrum ?-lactamase (ESBL-CTX-M), AmpC ?-Lactamase (CIT), Klebsiella pneumoniae carbapenemase (KPC) and New Delhi Metallo ?-lactamase (NDM) in Enterobacteriaceae colonising the gut of adult population in a South Indian rural community. Methodology: Faecal samples of 154 healthy volunteers were screened for Enterobacteriaceae resistant to commonly used antibiotics by standard methods, followed by phenotypic detection of ESBL by double disk synergy method, AmpC by spot inoculation and carbapenemases by imipenem and ethylenediaminetetraacetic acid + imipenem combined E-test strips and modified Hodge test. Polymerase chain reaction was done to detect blaCTX-M,blaCIT,blaKPC-1 and blaNDM-1 genes coding for ESBL, AmpC, KPC and NDM, respectively. Results: Colonisation rate of enteric bacteria with MDR genes in the community was 30.1%. However, phenotypically, only ESBL (3.2%) and NDM (0.65%) were detected. While the genes coding for ESBL, AmpC and NDM were detected in 35.6%, 17.8% and 4.4% of the MDR isolates, respectively. Conclusions: Carriage of MDR strains with a potential to express multidrug resistance poses a threat of dissemination in the community. Awareness for restricted use of antibiotics and proper sanitation can contain the spread of resistant bacteria.

Correlations between genotyping and antibiograms of clinical isolates of Pseudomonas aeruginosa from three different south Indian hospitals.

Purpose : To compare the molecular relationships and antibiograms of nosocomial isolates of Pseudomonas aeruginosa obtained from three different genres of hospitals located in Southern India, two located at Hyderabad (one private hospital and an ophthalmic hospital) and one in Puducherry (tertiary care teaching hospital). Each of these hospitals, which follow different infection control strategies and various problems associated with it, were investigated. Materials and Methods : Antibiograms generated by disk diffusion susceptibility testing for clinically relevant antibiotics and genotyping through fluorescent amplified fragment length polymorphism analysis (fAFLP) were the tools used in the study. Results: Molecular genotyping revealed a heterogeneous group of unrelated molecular clusters of P. aeruginosa strains having higher resistance that are apparently being endemic throughout the tertiary care teaching hospital. In eye care hospital, only a few distinct strains of P. aeruginosa predominating the study period were shown to be responsible for outbreaks. The third private hospital witnessed a group of resistant and persistent strains that might have clonally originated from a diverse collection of strains. Conclusions : The divergent kind of strains in our study suggests that there may be a direct link between the infection control practices followed in each hospital and kind of strains isolated in that particular setup. The study also emphasizes the need for maintaining infection control practices in hospitals with superior standards, failure of which might result in thriving of persistent P. aeruginosa clones in the hospitals.

Detection of pneumolysin and autolysin genes among antibiotic resistant Streptococcus pneumoniae in invasive infections.

Purpose: To detect the presence of autolysin and pneumolysin genes among Streptococcus pneumoniae strains isolated from different disease entities among Indian patients. The study also attempted to determine antimicrobial susceptibility of the isolates. Materials and Methods: A total of 24 S. pneumoniae isolates were checked for the presence of lytA gene coding for autolysin and ply gene coding for pneumolysin using polymerase chain reaction (PCR). All the isolates were subjected to susceptibility testing by disc diffusion method for 10 different therapeutically relevant antibiotics. Minimum inhibition concentration (MIC) was determined using broth dilution method for ampicillin, penicillin and ciprofloxacin. Results: Eleven isolates from ocular infections and 13 isolates from different invasive diseases showed susceptibility to most of the antibiotics tested except chloramphenicol and ciprofloxacin. Fifty percentage of the isolates showed resistance to chloramphenicol and ciprofloxacin. A moderate level of resistance of 18% was noted for cefepime and ceftriaxone. Only 6% of resistance was observed for amoxicillin and ceftazidime. MIC levels ranged from 0.015 to 1 μg/mL for ampicillin and only one isolate had an MIC of 1 μg/mL. The MIC levels for penicillin ranged from 0.062 to 4 μg/mL, wherein nine isolates showed high levels of MICs ranging from 2 to 4 μg/mL. Six isolates had a very high resistance levels for ciprofloxacin with MIC ranging from 32-128 μg/mL. The presence of lytA was observed in 23 out of 24 isolates tested whereas only 17 isolates were positive for pneumolysin. Four ocular isolates and one isolate from ear infection were negative for pneumolysin. Conclusion: Emerging resistance observed for cefepime and ceftriaxone might be due their increased and frequent usage nowadays. Presence of pneumolysin appears to be more critical for pathogenesis of invasive infections than the ocular infections. However, presence of lytA gene in all the isolates signifies that irrespective of site of isolation, kind of infection caused, autolysin is an obligate necessity for this organism.

Correlation between biofilm production and multiple drug resistance in imipenem resistant clinical isolates of Acinetobacter baumannii.

PURPOSE: To study the qualitative and quantitative methods for the investigation of biofilm formation and to examine the correlation between biofilm and antibiotic resistance among the clinical isolates of Acinetobacter baumannii . We also verified the association between biofilm and presence of extended spectrum beta-lactamases, particularly, bla PER-1 . METHODS: A total of 55 isolates were subjected to susceptibility testing by disc diffusion method for 13 clinically relevant antibiotics. Screening for biofilm production was done by both qualitative and quantitative methods through tube and microtitre plate assay respectively. The presence of bla PER-1 was checked by PCR. RESULTS: A. baumannii isolates showed very high resistance (>75%) to imipenem, cephotaxime, amikacin and ciprofloxacin. Only cefoperazone, netillin and norfloxacin were found to be effective agents. Results of microtitre and tube methods were concordant with 34 isolates (62%) showing biofilm formation. Resistance to four antibiotics such as amikacin (82% vs. 17.6%, P < 0.001), cephotaxime (88% vs. 11%, P P < 0.001), ciprofloxacin (70% vs. 29%, P =0.005) and aztreonam (38% vs. 11%, P =0.039) was comparatively higher among biofilm producers than non-biofilm producers. Microtitre assay additionally detected 14 weakly adherent isolates. Only 11 isolates had bla PER-1 gene and among these two were strong biofilm producers, while remaining were weakly adherent isolates. CONCLUSION: Microtitre plate method was found to be a more sensitive method for biofilm detection. This study demonstrates a high propensity among the clinical isolates of A. baumannii to form biofilm and a significant association of biofilms with multiple drug resistance. Presence of bla PER-1 appears to be more critical for cell adherence than for biofilm formation.

Characterization and susceptibility patterns of clinically important Enterococcus species in eastern Nepal.

Life threatening infections caused by enterococcus species with multidrug resistance has emerged as a threat to medical care in the present era. This study was conducted to characterize enterococcus species isolated from different clinical samples and to detect the pattern of susceptibility to some of the commonly used antibiotics in B.P Koirala Institute of Health Sciences (BPKIHS), a tertiary care hospital in eastern Nepal. Clinical samples submitted to the microbiology unit of Central Laboratory Service (CLS) for culture and sensitivity during March 2002 - February 2003 was analyzed. Enterococcus species were identified by colony characteristics, gram staining and relevant biochemical tests. Antibiotic susceptibility test was done by the Kirby Bauer disc diffusion technique. Of 50 Enterococcus species isolated, E. faecalis was the predominant isolate (48.0%) followed by E. faecium (32.0%) and E. avium (20.0%). Eighty-eight percent of E. faecalis showed sensitivity to cephotaxime and 87.0% to vancomycin. Multiple drug resistance was observed most commonly in E. faecium. Seventeen percent of E. faecium were resistant to vancomycin and 63.0% to ciprofloxacin and 44.0% to ampicillin. On the contrary E. avium rarely showed resistance to the antimicrobials tested including vancomycin. Enterococcal infections are common nowadays specially in hospitalized patients. Inappropriate use of antibiotics in clinical practice and poultry should be discouraged to prevent the emergence of multidrug resistant species.

A rare case of diphyllobothriasis from Pondicherry, South India.

Diphyllobothriasis is an intestinal parasitic infection caused by the ingestion of raw fresh-water fish containing the infectious larvae of Diphyllobothrium spp. This infection is uncommon in India. We report a case of diphyllobothriasis that occurred in Pondicherry, India, in a 5-year-old boy hailing from a fishing community. He attended the Pediatric OPD with spontaneous discharge of segments of the adult parasite. The segments (macroscopically and microscopically) were identified as those of Diphyllobothrium latum. The stool examination also revealed characteristic oval eggs.

Brain abscess due to Scedosporium apiospermum in a non immunocompromised child.

Scedosporium apiospermum is a filamentous fungi that commonly causes cutaneous infection. In certain circumstances, S. apiospermum can also cause invasive disease, which can involve the central nervous system (CNS). When the CNS becomes involved, treatment is difficult, therapeutic options are limited and the prognosis is poor. Early identification and treatment can decrease the mortality rate. Here we present a case of brain abscess with chronic suppurative otitis media, caused by S. apiospermum. This is the first such case report from Nepal. We could identify the organism only post mortem. We could not save the patient, probably due to delay in diagnosis.

Risk factors and associated problems in the management of infections with methicillin resistant Staphylococcus aureus.

PURPOSE: It is necessary to define the problem of methicillin resistant Staphylococcus aureus (MRSA) in every hospital to evolve control strategies. The objectives of this study were to determine factors influencing the persistence of MRSA in patients with hospital acquired infection and to identify alternate cost effective antibiotics. METHODS: A six month study was carried out for 50 patients with MRSA infection. Treatment modalities and risk factors were determined by a preset protocol. Minimum inhibitory concentration of commonly used antibiotics was determined. RESULTS: The risk factors were prolonged postoperative morbidity, prior antibiotic therapy and emergency admissions. Seventy percent of the isolates were from postoperative cases undergoing emergency surgeries. Isolation was highest during the second week of hospital stay. Emergency admissions had a significantly higher chance of early isolation. Prior treatment with multiple antibiotics in 38% was found to be another major risk factor. Ofloxacin was seen to be efficacious in a small percentage of cases. Rifampicin in combination with ofloxacin and clindamycin were found to be other good alternatives. Ofloxacin was found to be the cheapest and vancomycin the most expensive, for a full course of treatment. CONCLUSIONS: Minimizing risk factors and attention to alternate cost effective combination therapy may ease the problem of management of infections with MRSA.

Learning from an outbreak: ESBL- the essential points.

Extended spectrum beta-lactamase (ESBL) producing strains of Klebsiella pneumoniae have emerged as important nosocomial pathogens. The present report describes an outbreak of ESBL positive K.pneumoniae in a neonatal intensive care unit of a tertiary care center in southern India. The clinical and microbiological significance of multiresistant gram negative ESBL producers have been discussed in light of this outbreak by multi-resistant gram negative bacilli. The review also offers some practical guidance regarding infection control and therapeutic options.

Unusual morphological forms of cryptococcus neoformans in cerebrospinal fluid.

Cryptococcal meningitis is recognized as one of the AIDS defining conditions. It acts as a diagnostic marker for further investigations to establish the retroviral infection. Cryptococcus neoformans can be readily identified by its characteristic capsulated morphology. But rarely variations in morphology may be seen. We report a case of chronic meningitis in an AIDS patient whose cerebrospinal fluid (CSF) stained with India ink showed elliptical, peanut shaped and pseudomycelial large yeast cells with thick capsules. The organism was cultured and identified as Cryptococcus neoformans var neoformans. Unusual morphological forms may create diagnostic dilemma in the rapid diagnosis of cryptococcal meningitis.