Leishmaniasis is caused by the infection of haemoparasite Leishmania . The disease is a major public health problem in at least 88 countries, including India. Various species of Leishmania are involved in causing this disease. In India, Leishmania donovani species causes visceral leishmaniasis or kala-azar. The parasite is mainly transmitted from infected to uninfected person through the bites of female sandfly. Rarely the parasite can transmit through placenta from mother to child, through sexual intercourse, as laboratory acquired and through blood transfusion. This paper reports a unique case of transfusion-transmitted fatal kala-azar in an Indian infant who acquired this infection within few days of his birth after receiving blood from his maternal uncle, who was asymptomatic at the time of blood donation but died due to severe kala-azar within three months. The baby started having fever and developed hepatosplenomegaly within one month of blood transfusion and in spite of repeated anti-leishmanial treatment with sodium antimony gluconate the child died at the age of 7 months. This paper details the clinico-pathological findings of this child and also reviews the literature on this aspect and its impact on transfusion medicine.

Purpose: In resource-constrained laboratories of developing countries determination of antifungal susceptibility testing by NCCLS/CLSI method is not always feasible. We describe herein a simple yet comparable method for antifungal susceptibility testing. Methods: Reference MICs of 72 fungal isolates including two quality control strains were determined by NCCLS/CLSI methods against fluconazole, itraconazole, voriconazole, amphotericin B and cancidas. Dermatophytes were also tested against terbinafine. Subsequently, on selection of optimum conditions, MIC was determined for all the fungal isolates by semisolid antifungal agar susceptibility method in Brain heart infusion broth supplemented with 0.5% agar (BHIA) without oil overlay and results were compared with those obtained by reference NCCLS/CLSI methods. Results: Comparable results were obtained by NCCLS/CLSI and semisolid agar susceptibility (SAAS) methods against quality control strains. MICs for 72 isolates did not differ by more than one dilution for all drugs by SAAS. Conclusions: SAAS using BHIA without oil overlay provides a simple and reproducible method for obtaining MICs against yeast, filamentous fungi and dermatophytes in resource-constrained laboratories.

Purpose: AmpC producing K. pneumoniae have been increasingly reported from India but epidemiological studies are lacking. In the present study, molecular epidemiology of extended-spectrum AmpC beta-lactamases (ESACs) producing clinical isolates of K. pneumoniae prevalent in our hospital was studied. Methods: Fifty-one non-repeat, consecutive, clinical isolates of K. pneumoniae producing AmpC enzymes, were subjected to whole cell protein profile analysis (SDS-PAGE) and ribotyping. The antimicrobial susceptibility was determined using standard disk diffusion technique. The isolates showing decreased susceptibility to cefoxitin (<18 mm) or cefotetan (<16 mm) were subjected to modified three- dimensional test for detection of AmpC enzyme. Results: Six different types of protein profiles were observed. Ribotyping could further discriminate between the strains that were clustered by protein fingerprinting. Twelve different ribo-patterns were identified. Ribotyping was found to have a better Discriminatory Index (0.98) than that of SDS-PAGE (0.78). Of the 26 isolates that showed decreased susceptibility to cefoxitin and/or cefotetan 13 isolates were found to harbour AmpC enzyme. Conclusions: The study demonstrated the usefulness of SDS-PAGE whole cell protein profile analysis and ribotyping to identify the clonality of the ESACs isolates, the latter having a higher discriminatory power. The presence of ESACs isolates in the community as well as in hospital settings emphasizes the need for regular monitoring of antimicrobial resistance.

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.

Purpose: To study molecular profiles of oral Candida tropicalis isolates from five different geographic locales to determine the molecular diversity, clonality and evolutionary trends of this opportunistic pathogen. Methods: A total of 36 strains from five countries (China, Canada, Scotland, Japan and Tanzania) were genotyped by PCR fingerprinting with 11 separate primers. Of these, primers RSG9, RSG8, T3B and RSD12 generated complex fingerprinting patterns. Results: Three significantly dissimilar profiles were derived from the primer T3B and particularly focused on tDNA suggested the prevalence of genetic subtypes within the species. Comparison of tDNA and rDNA (RSD12) fingerprints of C. tropicalis suggested that rDNA is much more heterogeneous than the relatively distinct tDNA. Further analysis of similarity coefficient (SAB) of gel profiles derived from computer-generated dendrograms indicated some degree of similarity in isolates from five-disparate geographic locales as well as the presence of unique isotypes in each region. Conclusions: This study demonstrates the evolutionary divergence of distinct genetic subgroups within Candida tropicalis .

Purpose: Helicobacter pylori infection is common in the developing countries. The cagA gene is a marker of pathogenicity island (PAI) in H. pylori . The aim of this study was to determine the prevalence of cagA among dyspeptic patients in Bahrain directly from gastric biopsy and stool specimen. Methods: A total of 100 gastric biopsy samples, 16 clinical isolates and 44 faecal specimens were collected from Bahraini adult dyspeptic patients. cagA gene of H. pylori was assessed using polymerase chain reaction (PCR). Results: The cagA gene was detected in 59 (59%) from biopsy specimens, 10 (62%) clinical isolates and in 10 (22.7%) faecal specimens. The detection of cagA positive H. pylori was significantly higher in patients with duodenal ulcer (80%) compared to those with other endoscopic finding (42%) ( P <0.05). Conclusions: Using PCR to detect cagA gene directly from biopsy is a rapid and reliable technique. However, using stool specimen for genotyping in our patients showed reduced sensitivity.

An attempt was made to speciate 102 clinically significant isolates of coagulase negative staphylococci (CoNS) by a practical scheme adapted from various references. This scheme utilizes slide and tube coagulase test, urease test ornithine decarboxylase, novobiocin susceptibility and aerobic acid from mannose for assigning species group. Inclusion of one or two additional tests in a species group could identify the isolates to species level. Ninety eight (97%) isolates were conveniently identified as S. epidermidis (41%), S. saprophyticus (16.6%), S. haemolyticus (14.7%), S. hominis (14.7%), S. lugdunensis (4.9%), S. schleiferi (1.9%) and S. capitis (1.9%). Only four isolates were not identified to the species level, two of which were probably S. capitis subspecies ureolyticus / S. warneri / S. simulans . Antibiotic susceptibility testing showed maximum resistance to ampicillin (89%) followed by cefotaxime (59%) with no resistance to vancomycin. The increasing recognition of pathogenic potential of CoNS and emergence of drug resistance amongst them denotes the need to adopt simple laboratory procedures to identify and understand the diversity of staphylococci isolated from clinical material.

Escherichia coli isolates resistant to ceftazidime isolated in the University Malaya Medical Center (UMMC) Kuala Lumpur, Malaysia, between the years 1998 and 2000 were studied for extended-spectrum β -lactamase (ESBL) production. All strains were analysed phenotypically and genotypically and found to be ESBL-producing organisms harbouring SHV-5 β-lactamase. This was confirmed by PCR-SSCP and nucleotide sequencing of the blaSHV amplified gene. As there was no evidence of ESBL activity in E. coli prior to this, coupled with the fact that there was a predominance of SHV-5 β-lactamases in Klebsiella pneumoniae isolates in UMMC, we postulate that the E. coli obtained the SHV-5 β-lactamase genes by plasmid transfer from the ESBL-producing K. pneumoniae.

Increasing resistance to third generation cephalosporins has become a cause for concern especially among Enterobacteriaceae that cause nosocomial infections. The prevalence of extended spectrum β -lactamases (ESBLs) among members of Enterobacteriaceae constitutes a serious threat to current β -lactam therapy leading to treatment failure and consequent escalation of costs. A detailed study was initiated to identify the occurrence of ESBLs among the Enterobacteriaceae isolates at a tertiary care hospital using the double disk potentiation technique. Antibiogram profiles were determined to commonly used antibiotics and confirmation of ESBLs production was carried out by the disk diffusion assay using ceftazidime and cefotaxime in the presence and absence of clavulanic acid. Our results indicate that the majority of ESBLs were expressed in Escherichia coli.

The common cause of skin infections are dermatophytes and opportunistic fungi. Aim of this study was to isolate and identify the fungal agents from clinical samples from patients with different mycoses. Clinical samples from 165 patients were subjected to potassium hydroxide (KOH) examination and culture isolation; causative agents were identified macroscopically and microscopically. All the 165 specimens were KOH positive and 110/165 (66.7%) samples were culture positive. Of these, highest isolation rate was obtained in opportunistic mycoses such as candidiasis (29/29, 100%). Dermatophytes were isolated in 53/80 (66.3%) specimens and Trichophyton rubrum was the commonest isolate in skin samples (17/24) among the patients suffering from dermatophytosis. Phaeoannellomyces wernecki was isolated in a patient suffering from tinea nigra. The study signifies the importance of mycological examination in the diagnosis of various mycoses for their effective management.

A retrospective analysis of all cases of sporotrichosis was conducted to examine the pattern and frequency of sporotrichosis cases in Manipur over a period of six year from July 1999 to June 2005. Seventy-three (73) cases of sporotrichosis were detected of which 30 were confirmed by culture and 43 were diagnosed by aspiration cytology only. Most of the patients belonged to the age group of 21 to 40 years (n=23; 31.5%). There were 39 females (53.4%) and 34 males (46.5%). Commonest site of infection was upper limbs (n=39; 53.4%) followed by lower limbs (n=17; 23.2%). Commonest type of infection was lymphocutaneous form (n=46; 63.1%) followed by fixed cutaneous form (n=27; 36.9%). Among these 73 cases, 2 male patients (2.7%) were found to be HIV positive. In our experience, collection of material by aspiration of pus or infected tissue was found to be a better method compared to scraping or exudate. This study revealed Manipur as a new endemic area for sporotrichosis.

Angiostrongylus cantonensis is a nematode parasite that inhabits the pulmonary arteries and heart of rodents. It is one of the causative agents of fatal eosinophilic meningoencephalitis in man. We present five cases of eosinophilic meningitis presumably due to infection with Angiostrongylus cantonensis . All the five patients gave history of ingestion of monitor lizard within ten days of onset of symptoms.

We report a case of pulmonary zygomycosis in an adult male diabetic patient who presented with fever and altered sensorium initially and later developed streaky haemoptysis. Bronchoscopy showed picture of necrotizing pneumonia. Sputum was negative for fungal elements on admission but later bronchial wash and repeat sputum samples were positive by microscopy and culture showed growth of Rhizopus species. Immediately the patient was put on amphotericin B but had a bout of massive haemoptysis and succumbed. A high index of suspicion is needed for an early diagnosis and aggressive treatment of this infection in view of the high mortality rate.

Human immunodeficiency virus (HIV) and M. tuberculosis are two intracellular pathogens that interact with each other at both clinical and cellular levels. A known HIV positive case, presentingwith vague pulmonary symptoms followed by bilateral cervical lymphadenopathy, is reported here. The condition was treated as asthma, based on lung symptoms. Cytology revealed acute purulent lymphadenitis. Only Z. N. staining and culture of sputum and lymphnode aspirate could diagnose tuberculosis. Patient responded well to the standard anti tuberculosis treatment.

Cutaneous infections is observed in 15% of patients with disseminated cryptococcosis with AIDS. We present here a case of a 34 years old female with AIDS. She presented with multiple skin coloured umbilicated over face, neck, trunk and limbs, which mimicked molluscum contagiosum and kaposi sarcoma. The tissue from cutaneous lesions was collected by excision biopsy and processed by standard mycological methods. Cryptococcus neoformans was isolated and identified. Cerebrospinal fluid (CSF) also yielded the growth of C. neoformans . Cryptococcal antigen was detected with a titre of 1024 by Latex agglutination, is serum and CSF. Her serum was reactive for HIVI and 2 antibodies. The CD4 lymphocytes count was 80/cmm. The HIV viral load was 2,48,084 copies/mL. She was treated with amphotericin B injectable and oral fluconazole. She responded well and lesions regressed.

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.

Aspergilloma is a fungal ball that usually forms in a preformed stationary cavity, mostly in lung and paranasal sinuses. We report a rare case of primary orbital Aspergilloma following exenteration for an invasive ocular surface squamous neoplasia, clinically mimicking a recurrence of the tumor. The fungal ball showed the presence of conidiophores with a globular head and a complete row of uni and biserrate phialides, suggestive of Aspergillus flavus species. The exposure to air in the orbit, possibly promoted the formation of conidiophores, which are normally seen when the organism is located in air cavities.