Methicillin-Resistant Staphylococcus aureus Contamination of Frequently Touched Objects in Intensive Care Units: Potential Threat of Nosocomial Infections.

Background: Bacterial contamination in intensive care units is an important risk factor associated with increasing incidences of nosocomial infections. This study was conducted to study the bacterial colonization on commonly touched objects of intensive care units and antibiotic resistance pattern of bacterial isolates. Methods: This study was conducted in different intensive care units of Manipal Teaching Hospital, Pokhara, Nepal. A total of 235 swabs were collected from surfaces of bed rails, monitors, door handles, IV stands, telephone sets, nursing stations, medicine trolleys, sphygmomanometers, wash basin taps, dressing drums, stethoscopes, pulse oximeters, ventilators, defibrillators, and stretchers. Isolation, identification, and antibiotic susceptibility tests of the bacteria were performed following standard microbiological techniques. Results: Of 235 samples, bacterial growth was observed in 152 samples. A total of 90 samples of Staphylococcus aureus were isolated from 235 samples. Most of the sampling sites included in this study were found contaminated with S. aureus. The highest number of S. aureus was cultured from the surface of bed rails. Of the total S. aureus isolates, 54.4% (49/90) were methicillin-resistant Staphylococcus aureus (MRSA). Vancomycin resistance was detected among 8.1% MRSA isolates (4/49). Acinetobacter species were the commonest Gram-negative bacterial isolate. Conclusion: Bacterial contamination of the objects/instruments of the ICU was recorded to be high. The most common contaminating bacteria were S. aureus with a high percentage of MRSA and emergence of VRSA. Periodic microbiological surveillance, detection of contaminated sites, and effective decontamination methods would minimize the colonization by potential pathogens and their transmission.

Kerion Celsi in a Nepalese Boy: An Underdiagnosed Cause of Scalp Swelling.

Tinea capitis (TC) is an infection of the scalp and hair caused by a dermatophyte fungus. Typically caused by the zoophilic and geophilic species of Microsporum and Trichophyton, it remains the commonest cutaneous fungal infection in children. A 9-year-old Nepalese boy was evaluated in outpatients for multiple boggy scalp lesions for two months. Suspecting a bacterial etiology, the lesions were excised and sent for cultures. While bacterial cultures failed to grow, endothrix spores were readily detected in potassium hydroxide preparation and histopathology. Trichophyton tonsurans was identified by the phenotype method and later confirmed by matrix-assisted laser desorption ionization-time of flight (MALDI-TOF). Systemic antifungal therapy for 6 weeks along with local wound dressings resulted in complete recovery. At 2-year follow-up, focal alopecia was seen; however, no recurrence was noted.

Bacterial contamination of neonatal intensive care units: How safe are the neonates?

BACKGROUND: Intensive care units (ICU) are essential healthcare facility for life threatening conditions. Bacterial contamination of objects/instruments in ICU is an important source of nosocomial infections. This study is aimed to determine the level of bacterial contamination of instruments/objects which are commonly touched by healthcare workers and frequently come in contact with the neonates. METHODS: This hospital based prospective study was conducted in neonatal intensive care unit (NICU) of Manipal Teaching Hospital, Pokhara, Nepal. A total of 146 samples collected from surfaces of incubators, radiant warmers, suction tips, ventilators, stethoscopes, door handles, weighing machines, mothers' beds, phototherapy beds, laryngoscope, telephone sets, blood pressure machine, etc. formed the material of the study. Isolation, identification and antibiotic susceptibility of the bacterial isolates was performed by standard techniques. Blood culture isolates from NICU patients during the study period were compared with the environmental isolates. RESULTS: Out of 146 samples, bacterial growth was observed in 109. A total of 119 bacterial isolates were retrieved from 109 samples. Three common potential pathogens isolated were Escherichia coli (n = 27), Klebsiella species (n = 21) and Staphylococcus aureus (n = 18). Majority of E. coli and Klebsiella isolates were from incubators, suction tips and mothers' beds. Majority of S. aureus isolates were cultured from radiant warmers. Among S. aureus isolates, 33.3% (6/18) were methicillin resistant. Majority of the bacterial isolates were susceptible to gentamicin and amikacin. Common potential pathogens isolated from blood culture of NICU patients were S. aureus and Klebsiella species. CONCLUSION: High degree of bacterial contamination of objects/instruments in NICU was recorded. Isolation of potential pathogens like E. coli, Klebsiella species and S. aureus is a major threat of nosocomial infections. Blood culture data of NICU reflects possibility of nosocomial infections from contaminated sites. Gentamicin and amikacin may be used for empirical therapy in suspected cases of nosocomial infections in NICU.

Neonatal sepsis due to glycopeptide resistant Enterococcus faecium from colonized maternal gut- rare case evidence.

Background: Vancomycin-resistant enterococcal infections in the neonatal ICU are growing global problems. We report a case of neonatal septicemia by multidrug-resistant vancomycin-resistant Enterococcus faecium (VRE), the source of infection being the mother's gut. Case presentation: A newborn male child admitted to the neonatal intensive care unit (NICU) was diagnosed to have mild meconium aspiration syndrome, early onset neonatal septicemia, and bacteremia by multidrug and vancomycin-resistant Enterococcus faecium. Screening of gut flora of the baby and the mother were carried out to trace the source of infection. Stool cultures of the mother and the baby yielded Vancomycin-Resistant Enterococcus faecium. All three isolates of Enterococcus faecium had similar antibiogram, harbored the vanA gene and similar pulsed-field gel electrophoresis pattern. Baby responded to the 1 week therapy with oral linezolid suspension 20 mg/kg/day, 1 ml/t.d.s. No VRE was isolated from baby on a repeat stool culture 1 week after the linezolid therapy. He was discharged with the advice for the continuance of linezolid for seven more days. Conclusion: Isolation of MDR-VRE from the blood culture of the baby and stool specimens of the mother and the baby with the same antibiogram profile and clonal similarities reveals that maternal gut colonization was responsible for neonatal sepsis. Optimal infection control measures and the development of guidelines for monitoring VRE colonization in pregnant women might be useful in reducing the occurrence of neonatal sepsis.

Low rate of gut colonization by extended-spectrum ß-lactamase producing Enterobacteriaceae in HIV infected persons as compared to healthy individuals in Nepal.

A worldwide increase in the gastrointestinal colonization by extended-spectrum ß-lactamase (ESBL)-producing bacteria has been observed. Their prevalence amongst Healthy People Living with HIV (HPLWH) has not been investigated adequately. The aim of this study was to determine and compare the rates of and risk factors for intestinal carriage and acquisition of extended-spectrum ß-lactamase producing Enterobacteriaceae (ESBL-E) and carbapenemase-producing Enterobacteriaceae (CPE) among healthy people living with HIV (HPLWH) and healthy HIV negative population in the community. A cross-sectional study was conducted. Rectal swabs from HPLWH (n = 119) and HIV negative individuals (n = 357) from the community were screened for ESBL and CPE. Phenotypically confirmed ESBL-E strains were genotyped by multiplex PCR. The risk factors associated with ESBL-E colonization were analyzed by a multivariable conditional logistic regression analysis. Specimen from 357 healthy volunteers (213 female and 144 male) and 119 HPLWH (82 female and 37 male) with a median age of 30 [IQR 11-50] years were included in the study. ESBL colonization were found in 45 (37.82% [CI 29.09, 47.16]) and 246 (68.91% [CI 63.93, 73.49]), HPLWH and healthy HIV negative participants respectively. HPLWH had lower ESBL carriage rate (odds ratio 0.274 [CI 0.178, 0.423]) compared to healthy HIV negative subject's (p<0.01). In this study, no carbapenemase-producing bacteria were isolated.CTX-M-15 type was the most predominant genotype in both groups. Livestock contact and over-the-counter medications were significantly associated with a higher ESBL-E carriage rate among healthy subjects. This is the first study in Nepal that has demonstrated a high rate of gut colonization by ESBL-E in the community, predominantly of blaCTX-M-15 genotype. This study divulges the low fecal carriage rate of ESBL producing bacteria in HPLWH group compared to healthy individuals in western Nepal. The factors responsible for this inverse relationship of HIV status and gut colonization by ESBL-E are unidentified and require further large-scale study.

Bacterial contamination of frequently touched objects in a tertiary care hospital of Pokhara, Nepal: how safe are our hands?

Background: Objects frequently touched by patients and healthcare workers in hospitals harbor potential pathogens and may act as source of infectious agents. This study aimed to determine the bacterial contamination of common hospital objects frequently touched by patients, visitors and healthcare workers. Methods: A total of 232 samples were collected from various sites like surface of biometric attendance devices, elevator buttons, door handles, staircase railings, telephone sets and water taps. Isolation, identification and antibiotic susceptibility testing of the isolates was performed by standard microbiological techniques. Biofilm forming ability of the S. aureus isolates was tested by a microtitre plate method. Results: A total of 232 samples were collected and 219 bacterial isolates were recovered from 181 samples. Staphylococcus aureus was the most common bacterial isolate (44/219). Majority of S. aureus isolates were recovered from elevator buttons, biometric attendance devices and door handles. Among the S. aureus isolates, 36.3% (16/44) were methicillin resistant Staphylococcus aureus (MRSA) while remaining were methicillin sensitive Staphylococcus aureus (MSSA). Out of 44 S. aureus isolates, 12 (29.5%) were multidrug resistant and 14 (31.8%) were biofilm producers. The majority of MRSA isolates 62.5% (10/16) were biofilm producers. Acinetobacter was the most common Gram negative isolate followed by E coli and Pseudomonas species. Conclusions: High bacterial contamination of frequently touched objects with variety of potential pathogens and normal flora was detected. S. aureus was the most common bacterial isolate. Biofilm forming ability offers additional survival advantage to the organisms on these objects. Present study highlights the need of improved hand hygiene among healthcare workers and regular cleaning/disinfection of sites of frequent public contact.

Nasal and Pharyngeal Colonization by Bacterial Pathogens: A Comparative Study between Preclinical and Clinical Sciences Medical Students.

BACKGROUND: Upper respiratory tract is one of the commonest sites for microbial colonization. The colonized individuals are at risk of infections and can be a source of transmission of pathogens. Medical students are frequently exposed to a variety of infectious agents and more likely to get colonized by them. This study was aimed to determine the prevalence and to compare the colonization rates of nasal and pharyngeal bacterial pathogens among preclinical and clinical sciences medical students. METHODS: This cross-sectional study was conducted among 100 preclinical and 100 clinical sciences medical students. Isolation, identification, and antibiotic susceptibility testing of the isolates were performed by standard microbiological techniques. RESULTS: The nasal colonization by S. aureus and MRSA was 35% (70/200) and 19.5% (39/200), respectively. The nasal colonization by S. aureus and MRSA was significantly higher among clinical sciences students as compared to preclinical sciences students. Pharyngeal colonization by Haemophilus influenzae was significantly higher among clinical sciences students as compared to preclinical sciences students. The pharyngeal colonization by beta-hemolytic streptococci (nongroup A) was higher among preclinical sciences students than clinical sciences students. CONCLUSION: The nasal colonization by S. aureus and MRSA was higher among clinical sciences students. Pharyngeal colonization by potential bacterial pathogens was higher among clinical sciences students than preclinical students. Periodic screening of MRSA and potential throat pathogens of clinical sciences students and may reduce the incidences of nosocomial transmission of pathogens.

Potassium permanganate cleansing is an effective sanitary method for the reduction of bacterial bioload on raw Coriandrum sativum.

OBJECTIVE: Raw vegetables including flowers, leaves, stems, and roots are important carriers of food borne pathogens. We evaluated the bacteriological contamination of unwashed coriander leaves, and effectiveness of cleansing with 0.1% potassium permanganate solution as decontamination method. RESULTS: Significant bacterial contamination including pathogens like Salmonella species and Aeromonas species were isolated from unwashed coriander leaves. Decontamination with 0.1% potassium permanganate was found to be more effective than three steps wash with sterile water.

Antifungal susceptibility and phenotypic virulence markers of Candida species isolated from Nepal.

OBJECTIVE: Candida species are part of the commensal microflora in many anatomical sites of the human body; however, breach in the integrity of the body part and impaired immunity of the host can lead to invasive candidiasis. A number of virulence determinants could contribute towards its pathogenicity. Thus we attempted to evaluate the in vitro expression of different virulence factors among clinical isolates of Candida species and assayed their susceptibility patterns against a range of antifungal agents. RESULT: Of the total of 71 isolates we obtained, 48 (67.6%) were Candida albicans, 11 (15.49%) Candida tropicalis, 09 (12.67%) Candida glabrata and 03 (4.22%) were Candida krusei. Proteinase, phospholipase and esterase production could be revealed amongst 43 (60.56%), 44 (61.97%) and 49 (69.01%) isolates respectively. None of the isolates showed DNAase activity. Fifty-five (77.39%) isolates were biofilm producers, and 53 (74.6%) exhibited high cell surface hydrophobicity.

Burden of bacterial upper respiratory tract pathogens in school children of Nepal.

INTRODUCTION: Respiratory tract infections are one of the most common human infections in all age group and important cause of mortality and morbidity worldwide. Most bacterial upper respiratory tract infections are vaccine preventable. This study aimed to determine the prevalence of carrier state of bacterial upper respiratory tract pathogens among school children. It also aimed to study their antibiograms. METHODS: The specimen from posterior pharyngeal wall and tonsils were collected from 204 participants on calcium alginate coated swabs (HiMedia). Isolates were identified by standard microbiological methods and tested for in vitro antibiotic susceptibility testing by modified Kirby-Bauer disc diffusion method. RESULTS: In this study, Streptococcus pneumoniae (16.6%) was the most common bacterial pathogen recovered, followed by Staphylococcus aureus (14.7%), ß-haemolytic streptococci (non-Group A) (8.8%), Streptococcus pyogenes (5.3%) and Corynebacterium diphtheriae (3.4%). The Gram negative bacteria were Klebsiella pneumoniae (4.9%), Haemophilus influenzae (3.4%) and Neisseria meningitidis (1.4%). Important findings in antibiogram include high resistance of Streptococcus pneumoniae to penicillin (91.17%) and resistance of S. aureus to oxacillin (23.3%). CONCLUSION: Pharyngeal colonisation by S. pneumoniae was found high among school children and this calls for an urgent need to include pneumococcal vaccine in routine national immunisation schedule of Nepal given the high burden of invasive pneumococcal disease. Despite expected universal vaccination, pharyngeal colonisation by C. diphtheriae is possible and there is possibility of transmission of these respiratory pathogens to other healthy children.

Diversity, in-vitro virulence traits and antifungal susceptibility pattern of gastrointestinal yeast flora of healthy poultry, Gallus gallus domesticus.

BACKGROUND: Poultry farming and consumption of poultry (Gallus gallus domesticus) meat and eggs are common gastronomical practices worldwide. Till now, a detailed understanding about the gut colonisation of Gallus gallus domesticus by yeasts and their virulence properties and drug resistance patterns in available literature remain sparse. This study was undertaken to explore this prevalent issue. RESULTS: A total of 103 specimens of fresh droppings of broiler chickens (commercial G domesticus) and domesticated chickens (domesticated G domesticus) were collected from the breeding sites. The isolates comprised of 29 (33%) Debaryozyma hansenii (Candida famata), 12 (13.6%) Sporothrix catenata (C. ciferrii), 10 (11.4%) C. albicans, 8 (9.1%) Diutnia catenulata (C. catenulate), 6 (6.8%) C. tropicalis, 3 (3.4%) Candida acidothermophilum (C. krusei), 2 (2.3%) C. pintolopesii, 1 (1.1%) C. parapsilosis, 9 (10.2%) Trichosporon spp. (T. moniliiforme, T. asahii), 4 (4.5%) Geotrichum candidum, 3 (3.4%) Cryptococcus macerans and 1 (1%) Cystobasidium minuta (Rhodotorula minuta). Virulence factors, measured among different yeast species, showed wide variability. Biofilm cells exhibited higher Minimum Inhibitory Concentration (MIC) values (µg/ml) than planktonic cells against all antifungal compounds tested: (fluconazole, 8-512 vs 0.031-16; amphotericin B, 0.5-64 vs 0.031-16; voriconazole 0.062-16 vs 0.062-8; caspofungin, 0.062-4 vs 0.031-1). CONCLUSIONS: The present work extends the current understanding of in vitro virulence factors and antifungal susceptibility pattern of gastrointestinal yeast flora of G domesticus. More studies with advanced techniques are needed to quantify the risk of spread of these potential pathogens to environment and human.

Onychomycosis due to Candida parapsilosis in a Child with Ventricular Septal Defect: An Unusual Predisposition.

Candida parapsilosis is emerging as a potential pathogen for onychomycosis. A 4-year-old male child with perimembranous ventricular septal defect (VSD) was admitted with features of cystitis and was treated with broad spectrum antibiotics. Two weeks later, he developed yellowish discoloration of nails of both hands. The sloughed out nail, on microscopy, showed numerous yeast forms that were identified as Candida parapsilosis by both phenotypic and genotypic methods. Antifungal sensitivity testing of the isolate was performed by microbroth dilution method in accordance with CLSI guidelines. Patient was successfully treated with topical amphotericin B and oral fluconazole. Thus, one should have a high index of suspicion of C. parapsilosis onychomycosis, especially when the patient is in the paediatric age group, presenting with unusual predisposing condition like congenital heart disease, and is on broad spectrum antibiotics.

Association of Panton Valentine Leukocidin (PVL) genes with methicillin resistant Staphylococcus aureus (MRSA) in Western Nepal: a matter of concern for community infections (a hospital based prospective study).

BACKGROUND: Methicillin resistant Staphylococcus aureus (MRSA) is a major human pathogen associated with nosocomial and community infections. Panton Valentine leukocidin (PVL) is considered one of the important virulence factors of S. aureus responsible for destruction of white blood cells, necrosis and apoptosis and as a marker of community acquired MRSA. This study was aimed to determine the prevalence of PVL genes among MRSA isolates and to check the reliability of PVL as marker of community acquired MRSA isolates from Western Nepal. METHODS: A total of 400 strains of S. aureus were collected from clinical specimens and various units (Operation Theater, Intensive Care Units) of the hospital and 139 of these had been confirmed as MRSA by previous study. Multiplex PCR was used to detect mecA and PVL genes. Clinical data as well as antimicrobial susceptibility data was analyzed and compared among PVL positive and negative MRSA isolates. RESULTS: Out of 139 MRSA isolates, 79 (56.8 %) were PVL positive. The majority of the community acquired MRSA (90.4 %) were PVL positive (Positive predictive value: 94.9 % and negative predictive value: 86.6 %), while PVL was detected only in 4 (7.1 %) hospital associated MRSA strains. None of the MRSA isolates from hospital environment was found positive for the PVL genes. The majority of the PVL positive strains (75.5 %) were isolated from pus samples. Antibiotic resistance among PVL negative MRSA isolates was found higher as compared to PVL positive MRSA. CONCLUSION: Our study showed high prevalence of PVL among community acquired MRSA isolates. Absence of PVL among MRSA isolates from hospital environment indicates its poor association with hospital acquired MRSA and therefore, PVL may be used a marker for community acquired MRSA. This is first study from Nepal, to test PVL among MRSA isolates from hospital environment.

Streptococcus pneumoniae and Haemophilus species colonization in health care workers: the launch of invasive infections?

BACKGROUND: Streptococcus pneumoniae and Haemophilus influenzae are important human pathogens. The risk of airborne and droplet-transmitted respiratory tract infections in healthcare workers (HCW) is substantial. The aim of this study was to determine the extent of oropharyngeal colonization with S. pneumoniae and Haemophilus spp. their antibiogram and risk factors of colonization in HCW at a tertiary care center, Western Nepal. METHODS: During 3 month period, 100 oropharyngeal swab specimens were collected from HCW of Manipal Teaching Hospital and 50 from non HCW from community. All the 150 specimens were screened for Haemophilus spp. and S. pneumoniae by standard techniques. Serotyping of H. influenzae type b was done by using specific antiserum. Antibiotic sensitivity patterns of isolates were determined by modified Kirby Bauer disc diffusion method. Association between the groups was analyzed using the Pearson χ(2) test and Fisher exact test. A forward step logistic regression model was used to identify significant predictors for colonization. RESULT: Sixty-five percent of HCW were colonized with S. pneumoniae and/or Haemophilus species compared to 32 % of non-HCW. Health care workers had odd ratio (OR) 3.946 [CI (1.916, 8.128)] times more tendency of colonization compared to non-HCW (P < 0.05). Pneumococcal colonization was observed high among smokers (81.5 %). Amongst HCW, post graduate resident doctors had higher rate of colonization (83.3 %) followed by interns (64.9 %), least being amongst the laboratory workers (58.3 %). CONCLUSION: The higher rate of colonization amongst HCW raises the possibility of occupational risk as well as horizontal spread of infections.

Emergence of Magnusiomyces capitatus infections in Western Nepal.

Magnusiomyces capitatus is an emerging opportunistic yeast in the Mediterranean region. We report from Nepal one case of M. capitatus infection and six other cases of colonization/probable infection due to M. capitatus at a tertiary care center. Majority of the patients were immunocompromised, at extreme age, associated with comorbidities, and had history of close contact with livestock and poultry. The isolates were identified by phenotypic and genotypic (ITS and D1/D2 region of 26S rDNA sequence) methods. Molecular typing of the isolates was carried out by amplified fragment length polymorphism. Minimum inhibitory concentration (MIC) of the isolates for amphotericin B, caspofungin, fluconazole, itraconazole, voriconazole, posaconazole, anidulafungin, and micafungin were 2, 0.1-4, 2, 0.12-0.5, 0.12-0.5, 0.25, 1-4, and 1-4 µg/ml, respectively. Presence of M. capitatus infection was not known in Nepal, and the study should alert the clinicians and infectious disease specialists.

Hepatitis B Virus (HBV) and Hepatitis C Virus (HCV) Co-Infection Among HIV Infected Individuals at Tertiary Care Hospital in Western Nepal.

BACKGROUND: The HIV, HBV and HCV viruses are the major public health concern all over the world including Nepal. The aim of the study is to determine the rate of HBV and HCV co-infections in patients with HIV infection. METHODS: The study cohort included 218 consecutive HIV infected patients who were examined for co-infection with HBV or HCV or both at Manipal teaching hospital, Western Nepal. The demographic data of the subjects was collected retrospectively. The data was analyzed with SPSS software and EPI Info to measure the correlation of variables and infection rates. RESULTS: In the course of six years study period, a total of 25,708 samples were collected for HIV screening test. The 218 (0.8%) screen test positive for HIV were confirmed as per WHO guidelines. The overall rate of co-infection with HBV and or HCV was 7.3% (16 of 218 patients). Only 7 (3.2% [CI 1.3, 6.5]) were positive for both HIV and HBV infection markers and 9 (4.1% [CI 1.9, 7.7]) were positive for HIV and HCV infection markers. None were positive of all three virus markers. CONCLUSION: It is advisable to implementregular screening for Hepatitis B Virus and Hepatitis C Virus among all HIV infected individuals and their sexual partners.