Epidemiological and clinico-mycological profile of fungal wound infection from largest burn centre in Asia.

The current study was conducted to know the incidence, predisposing factors, spectrum, clinical profile and antifungal susceptibility (AFS) of fungal wound infection (FWI) in burn patients. Of a total of 71 patients, 20 (28.2%) emerged with the diagnosis of FWI. Fungal pathogens in this study were Candida tropicalis (14%), Candida parapsilosis (5.6%), Aspergillus niger (2.8%) and one each of Candida albicans (1.4%), Candida glabrata (1.4%), Syncephalestrum (1.4%) and Fusarium solani (1.4%). All patients with mould infections expired before the mycological culture results could be conveyed to clinicians. Of the yeasts isolated in the study, one each of C. tropicalis and C. albicans showed cross-resistance to azoles. All the moulds were susceptible to amphotericin B. This study depicted that fungal invasion is associated with a high mortality, burn size 30-60% and high incidence of inhalational injury. Fungal invasion was detected on an average of 14 days after injury. Association of use of four classes of drugs - aminoglycosides, imipenem, vancomycin and third generation cephalosporins and use of total parenteral nutrition was observed. Expedient laboratory diagnosis of FWI and appropriate systemic antifungal therapy guided by AFS may improve outcome for severely injured burn victims.

Sporotrichosis in Delhi among the migrant population from Uttarakhand, India.

Six cases of sporotrichosis in Delhi (non-endemic area) were recorded in migrants from Uttarakhand over a period of one year (2008-2009). The clinico-epidemiological and mycological profile of the patients is discussed. These were confirmed by mycological culture, histopathological examination and/or remission of lesion with potassium iodide. Sporotrichosis is endemic in Uttarakhand, a north-western state in the sub-Himalayan region.

Determination of methicillin resistant Staphylococcus aureus (MRSA) using several phenotypic methods--a report from a tertiary care center at New Delhi, India.

Staphylococcus aureus is a serious current health care concern. Rapid, precise identification of MRSA is a prerequisite tool for control of hospital infection. The present study aimed at studying the antibiotic susceptibility and demographic profile of MRSA isolated from clinical specimens and also to assess the reliability of five methods of identifying methicillin resistance. A total of 112 isolates of Staphylococcus aureus isolated from critical areas of the hospital were randomly selected and subjected to various phenotypic methods for determining methicillin resistance. Determination of minimum inhibitory concentration (MIC) for oxacillin by microbroth dilution method was the 'gold standard'. The present study also focuses on the comparison of oxacillin disc diffusion and cefoxitin disc diffusion screening for determination of MRSA. The sensitivity and specificity values for latex agglutination, oxacillin salt agar screening, E-Strip, cefoxitin disc diffusion and oxacillin disc diffusion method are 98.7% and 72.5%, 94.4% and 92.5%, 93.1% and 65.0%, 86.1% and 65%, 83.3% and 90.0% respectively. No single phenotypic test is completely reliable for the detection of oxacillin resistance in S. aureus. Oxacillin salt agar screening at 6 microg/ml and PBP2' detection by latex agglutination method were the most sensitive and specific method for detecting MRSA. Amidst the disc screening methods which are most often used in the smaller laboratory set-up where agar screen may not be functionally feasible for various reasons (economic and performance) cefoxitin disc screening has good sensitivity and specificity overall and perhaps is closest to being the cheap and reliable alternative in these settings.

Standardization of first and second-line antitubercular susceptibility testing using BacT Alert 3D system: a report from a tertiary care centre in India

Resurgence of multidrug resistant tuberculosis has lead to demand for rapid susceptibility testing. Conventional methods take > 3 weeks and are tedious. Automated methods have superseded them for first line drug susceptibility testing. An attempt was made to standardize first and second line susceptibility testing using the BacT Alert 3D system (Biomerieux). And compare results with Lowenstein Jensen's (LJ) method. 121 isolates of Mycobacterium tuberculosis, 67 pulmonary and 54 extra pulmonary were subjected to sensitivity to first and second line drugs. Multidrug resistance was detected equally by both methods at 15.7 percent. 100 percent agreement was observed between the two methods for aminoglycosides, rifampicin, ethionamide and ciprofloxacin. 91.5 percent agreement was observed for isoniazid, 85 percent for pyrazinamide and 72.4 percent for ethambutol. The time taken by LJ method was 18-32 days and BacT Alert 3D system took 4-12 days. In the lesser developed nations where tuberculosis is rampant a rapid effective method for confirming multidrug resistant tuberculosis is definitely desirable and the BacT Alert 3D system was found an effective method when compared to the 'gold standard' LJ proportion.

Rhinosporidiosis in Delhi, north India: case series from a non-endemic area and mini-review.

We report three cases of rhinosporidiosis from migrant population of Delhi. Three male patients had sino-nasopharyngeal, nasopharyngeal and nasal rhinosporidiosis, respectively. One patient gave a history of bathing in stagnant water. The diagnosis was made by clinical presentation and microscopic observation of characteristic sporangia of Rhinosporidium seebri in mycological and histopathological investigations. All the patients were successfully treated with complete surgical excision of lesions and cauterization of base. There were no recurrences.

In vitro evaluation of a new cefixime-clavulanic acid combination for gram-negative bacteria.

The study was conducted to evaluate a new cefixime-clavulanic acid combination for in vitro susceptibility towards gram-negative bacteria. A total of 220 isolates of Escherichia coli, Klebsiella pneumoniae, Pseudomonas aeroginosa, Acinetobacter spp, Salmonella enterica serovar Typhi and Salmonella enterica serovar Typhimurium were included in the study. The isolates were tested for susceptibility towards the new combination antimicrobial molecule cefixime with clavulanic acid by disk diffusion and Epsilometer strip (E-strip) Minimum Inhibitary Concentration (MIC) method. Of the 101 E. coli and K. pneumoniae isolates, 62.4% were found to be extended spectrum beta-lactamase (ESBL) producers. Almost half of these were from the community and 55.6% were hospital isolates. Of the ESBL isolates, 19% were AmpC (cephalosporinases that are poorly inhibited by beta lactamase inhibitor) producers while the remaining 81% were non AmpC ESBL producers. The AmpC producers were resistant to both cefixime and the combination, while the non-AmpC producers were sensitive to the combination. The addition of clavulanate to cefixime did not improve the sensitivities of P. aeruginosa and Acinetobacter isolates. There were no ESBL isolates among the S. Typhi isolates, all of which were sensitive to cefixime. Of the S. Typhimurium, 88.9% were ESBL producers and all of these were resistant to cefixime but sensitive to the combination. The combination of cefixime with clavulanic acid offers the advantage of oral administration and appears to be a viable option for the treatment of uncomplicated community acquired infections caused by non-AmpC ESBL producing gram-negative bacteria.

Minimum inhibitory concentration of carbapenems and tigecycline against Salmonella spp.

Antimicrobial resistance in Salmonella spp. is of grave concern, more so in quinolone-resistant and extended-spectrum beta-lactamase (ESBL)-producing isolates that cause complicated infections. The MIC of azithromycin, ciprofloxacin, cefixime, cefepime, ceftriaxone, gatifloxacin, imipenem, levofloxacin, meropenem and ofloxacin (E-test strip) and tigecycline and faropenem (agar dilution) against 210 Salmonella spp. was determined. MIC(90) (defined as the antimicrobial concentration that inhibited growth of 90 % of the strains) of the carbapenems (imipenem and meropenem) for Salmonella Typhi and Salmonella Paratyphi A was 0.064 microg ml(-1). MIC(90) of faropenem was 0.25 microg ml(-1) for S. Typhi, S. Paratyphi A and Salmonella Typhimurium. The MIC(90) of azithromycin for all Salmonella spp. ranged from 8 to 16 microg ml(-1). Tigecycline showed an MIC(90) of 2 microg ml(-1) for S. Typhi, 1 microg ml(-1) for S. Paratyphi A and 4 microg ml(-1) for S. Typhimurium. We concluded that tigecycline and the carbapenems are likely to have roles in the final stage of treatment of quinolone-resistant and ESBL-producing multidrug-resistant salmonellae.

Standardization of first and second-line antitubercular susceptibility testing using BacT Alert 3D system: a report from a tertiary care centre in India.

Resurgence of multidrug resistant tuberculosis has lead to demand for rapid susceptibility testing. Conventional methods take > 3 weeks and are tedious. Automated methods have superseded them for first line drug susceptibility testing. An attempt was made to standardize first and second line susceptibility testing using the BacT Alert 3D system (Biomerieux). And compare results with Lowenstein Jensen's (LJ) method. 121 isolates of Mycobacterium tuberculosis, 67 pulmonary and 54 extra pulmonary were subjected to sensitivity to first and second line drugs. Multidrug resistance was detected equally by both methods at 15.7%. 100% agreement was observed between the two methods for aminoglycosides, rifampicin, ethionamide and ciprofloxacin. 91.5% agreement was observed for isoniazid, 85% for pyrazinamide and 72.4% for ethambutol. The time taken by LJ method was 18-32 days and BacT Alert 3D system took 4-12 days. In the lesser developed nations where tuberculosis is rampant a rapid effective method for confirming multidrug resistant tuberculosis is definitely desirable and the BacT Alert 3D system was found an effective method when compared to the 'gold standard' LJ proportion.

Microflora of bile aspirates in patients with acute cholecystitis with or without cholelithiasis: a tropical experience.

The current study determined the spectrum of biliary microflora with special emphasis on enteric fever organisms in patients with acute cholangitis with and without cholelithiasis or other biliary diseases. The patients were divided into three groups: Group A consisted of patients with acute cholecystitis with cholelithiasis; Group B consisted of patients with acute cholecystitis with gastrointestinal ailments requiring biliary drainage and group C consisted of patients with gallbladder carcinoma. Gallbladder, bile and gallstones were subjected to complete microbiological and histopathological examination. Antimicrobial susceptibility of the isolates was performed as per CLSI guidelines. Bacteria were recovered from 17 samples (32%) in Group A, 17 (51.4%) in Group B and 1 (1.6%) in Group C. The most common organisms isolated were Escherichia coli (11, 29.7%), Klebsiella pneumoniae (10, 27%), Citrobacter freundii (3, 8.1%), Salmonella enterica serovar Typhi (3, 8.1%), etc. The majority of Enterobacteriaceae isolates were susceptible to piperacillin-tazobactam and meropenem. As regards Salmonella spp., S. Typhi was isolated from 2 (3.8%) patients in Group A and 1 (16%) in Group C. Antimicrobial susceptibility of potential causative organisms, the severity of the cholecystitis, and the local susceptibility pattern must be taken into consideration when prescribing drugs. A protocol regarding the management of such cases should be formulated based on observations of similar studies.

Neurocysticercosis in a north Indian hospital.

In endemic regions, neurocysticercosis (NCC) is the most commonly diagnosed parasitic disease of the central nervous system, and the most common cause of convulsions and hydrocephalus in adults. During January 2000-December 2006, serum samples collected from patients presenting with various manifestations with a clinical diagnosis of cysticercosis and/or relevant computed tomography findings were subjected to an enzyme-linked immunosorbent assay test for NCC. Anti-cysticercus antibodies were detected in 155 of the 1096 (14.1%) cases. Generalized seizure (33.9%) was the most common presenting symptom. Solitary lesion (74.2%) was the most common radiological finding. This study provides an assessment of the epidemiology of NCC in Delhi and stresses the need for its prevention.

Microflora of bile aspirates in patients with acute cholecystitis With or without cholelithiasis: a tropical experience

The current study determined the spectrum of biliary microflora with special emphasis on enteric fever organisms in patients with acute cholangitis with and without cholelithiasis or other biliary diseases. The patients were divided into three groups: Group A consisted of patients with acute cholecystitis with cholelithiasis; Group B consisted of patients with acute cholecystitis with gastrointestinal ailments requiring biliary drainage and group C consisted of patients with gallbladder carcinoma. Gallbladder, bile and gallstones were subjected to complete microbiological and histopathological examination. Antimicrobial susceptibility of the isolates was performed as per CLSI guidelines. Bacteria were recovered from 17 samples (32 percent) in Group A, 17 (51.4 percent) in Group B and 1 (1.6 percent) in Group C. The most common organisms isolated were Escherichia coli (11, 29.7 percent), Klebsiella pneumoniae (10, 27 percent), Citrobacter freundii (3, 8.1 percent), Salmonella enterica serovar Typhi (3, 8.1 percent), etc. The majority of Enterobacteriaceae isolates were susceptible to piperacillin-tazobactam and meropenem. As regards Salmonella spp., S. Typhi was isolated from 2 (3.8 percent) patients in Group A and 1 (16 percent) in Group C. Antimicrobial susceptibility of potential causative organisms, the severity of the cholecystitis, and the local susceptibility pattern must be taken into consideration when prescribing drugs. A protocol regarding the management of such cases should be formulated based on observations of similar studies.

Epidemiologic and clinicomycologic profile of onychomycosis from north India.

BACKGROUND: Onychomycosis is an important public health problem because of the increase in immunosuppressive states. Large-scale studies in India are scarce, and so the baseline incidence of onychomycosis is not firmly established. METHODS: Three hundred and two clinically suspected cases of onychomycosis were included in this study. Nail samples were collected for direct microscopic examination and culture. Clinical patterns and associated relevant factors were noted according to a predetermined protocol. RESULTS: The associated predisposing conditions included diabetes mellitus (3.9%) and systemic lupus erythematosus (2.3%). Distal and lateral subungual onychomycosis was the most common clinical pattern (62%), followed by total dystrophic onychomycosis (20.2%). The most common fungal isolates were dermatophytes (49.5%), followed by Candida spp. (40.4%) and nondermatophyte molds (10.1%). Of the dermatophytes, Trichophyton rubrum (47%) was the most common isolate, followed by Trichophyton tonsurans (20.4%). Of the Candida spp., Candida albicans was the most common (60%). CONCLUSIONS: Until recently, yeasts and nondermatophyte molds were regarded as contaminants, but their emergence as a significant cause of onychomycosis in immunocompromised patients calls for mycologic diagnosis and antifungal susceptibility testing in onychomycosis. The recognition of the changing patterns of onychomycosis will aid in the therapeutic approach and the implementation of control measures.

Current scenario of cryptococcosis and antifungal susceptibility pattern in India: a cause for reappraisal.

This study analysed the spectrum, antifungal susceptibility pattern, clinical course and molecular epidemiology of cryptococcosis. Four hundred and thirty-nine samples obtained from 378 meningitis patients were processed by standard procedures. Minimum inhibitory concentration (MIC) of fluconazole and amphotericin B for the isolates was tested by broth micro dilution and by E-strip method. Molecular analysis by random amplified polymorphic DNA-PCR of eight isolates was performed using M13 primer. Cryptococcosis was diagnosed in 35 patients [HIV-1 seropositive (19) and apparently immunocompetent (16)]. Cryptococcus neoformans var. neoformans (serotype A and D) was the predominant isolate on phenotypic identification. Three C. neoformans var. gattii were isolated from HIV-1 seropositive (2) and apparently immunocompetent (1) patients. MIC 90 for amphotericin B and fluconazole were 1 and 8 mug ml(-1) respectively. On RAPD-PCR, less diversity was seen among Indian isolates. AIDS remains the single most important risk factor for cryptococcosis. Rising MIC of the available induction and maintenance drugs is of grave concern. The DNA typing technique showed less diversity among Indian strains. Routine surveillance and application of molecular typing methods are crucial to know the baseline and existing pattern of cryptococcosis.

Disseminated cryptococcosis with necrotizing fasciitis in an apparently immunocompetent host: a case report.

We present an unusual case of disseminated cryptococcosis with necrotizing fascitis in an immunocompetent host. The multi-system cryptococcal infection was proved using standard microbiological, histopathological and radiological investigations. After a combination of antifungal, antimicrobial and surgical therapy clinical cure was achieved.

In vitro activity of azithromycin, newer quinolones and cephalosporins in ciprofloxacin-resistant Salmonella causing enteric fever.

The therapeutic alternatives available for use against ciprofloxacin-resistant enteric fever isolates in an endemic area are limited. The antibiotics currently available are the quinolones, third-generation cephalosporins and conventional first-line drugs. In this study, the MICs of various newer drugs were determined for 31 ciprofloxacin-resistant enteric fever isolates (26 Salmonella enterica serovar Typhi and 5 S. enterica serovar Paratyphi A). MICs for ciprofloxacin, ofloxacin, gatifloxacin, levofloxacin, cefotaxime, cefixime, cefepime and azithromycin were determined using Etest strips and the agar dilution method. By Etest, all of the ciprofloxacin-resistant isolates had ciprofloxacin MICs >/=32 mug ml(-1). S. Typhi showed MIC(90) values of 0.50, 0.25 and 0.38 mug ml(-1) for cefixime, cefotaxime and cefepime, respectively. For the cephalosporins, a negligible difference in MIC(90) and MIC(50) values for S. Typhi and S. Paratyphi A was observed. A single isolate of S. Typhi showed a high azithromycin MIC of 64 mug ml(-1). The MIC(90) value for azithromycin in S. Typhi and S. Paratyphi was 24 mug ml(-1). Gatifloxacin demonstrated lower resistance (80.8 %) compared with the other quinolones (92-100 %) in S. Typhi. The rise in MIC levels of these antimicrobials is a matter for serious concern.

Salmonella enterica serovar typhi: molecular analysis of strains with decreased susceptibility and resistant to ciprofloxacin in India from 2001-2003.

Chromosomally-mediated reduced susceptibility to ciprofloxacin narrows the therapeutic options in enteric fever. We made a molecular comparison of clinical isolates of fluoroquinolone-resistant strains of Salmonella enterica serotype Typhi from January 2001 to May 2003; 178 isolates were subjected to antimicrobial susceptibility testing by the Kirby-Bauer method of disk diffusion, and agar dilution was used to determine the minimum inhibitory concentration (MIC) to ciprofloxacin. Nalidixic-acid resistant strains (NARST) were observed in 51% of the isolates, of which 98.9% had decreased susceptibility (MIC> or =0.125-1 microg/mL) to ciprofloxacin. A single strain (4 microg/mL) was resistant to ciprofloxacin and double mutations were found in the gyrA gene (76 Asp->Asn, 44 leu->Ileu). Among seven NARST strains with reduced susceptibility, a single mutation was found in five strains, one of which had 76 Asp->Asn and two each had mutations at 87 Asp->Asn and 72 Phe->Tyr, respectively); no mutations could be detected in two isolates. Routine antimicrobial surveillance, coupled with molecular analysis of fluoroquinolone resistance, is crucial for revision of enteric fever therapeutics.

Salmonella enterica serovar typhi: molecular analysis of strains with decreased susceptibility and resistant to ciprofloxacin in india from 2001-2003

Chromosomally-mediated reduced susceptibility to ciprofloxacin narrows the therapeutic options in enteric fever. We made a molecular comparison of clinical isolates of fluoroquinolone-resistant strains of Salmonella enterica serotype Typhi from January 2001 to May 2003; 178 isolates were subjected to antimicrobial susceptibility testing by the Kirby-Bauer method of disk diffusion, and agar dilution was used to determine the minimum inhibitory concentration (MIC) to ciprofloxacin. Nalidixic-acid resistant strains (NARST) were observed in 51 percent of the isolates, of which 98.9 percent had decreased susceptibility (MIC>0.125-1mug/mL) to ciprofloxacin. A single strain (4 mug/mL) was resistant to ciprofloxacin and double mutations were found in the gyrA gene (76 Asp->Asn, 44 leu->Ileu). Among seven NARST strains with reduced susceptibility, a single mutation was found in five strains, one of which had 76 Asp->Asn and two each had mutations at 87 Asp->Asn and 72 Phe->Tyr, respectively); no mutations could be detected in two isolates. Routine antimicrobial surveillance, coupled with molecular analysis of fluoroquinolone resistance, is crucial for revision of enteric fever therapeutics.

Ciprofloxacin-resistant Neisseria meningitidis, Delhi, India.

Decreased susceptibility of Neisseria meningitidis isolates to ciprofloxacin emerged from an outbreak in Delhi, India. Results of antimicrobial susceptibility testing of the meningococcal isolates to ciprofloxacin and further sequencing of DNA gyrase A quinolone-resistance-determining region confirmed the emergence of ciprofloxacin resistance in the outbreak.

Resistance to erythromycin and rising penicillin MIC in Streptococcus pyogenes in India.

There has been worldwide resurgence in the incidence of Streptococcus pyogenes infection and its sequelae. S. pyogenes remains uniformly susceptible to penicillin, and it is speculated that its minimum inhibitory concentration (MIC) has not changed during the past 70 years. The purpose of the present study was to determine the occurrence and pattern of resistance to penicillin and erythromycin amongst clinical isolates of S. pyogenes. A total of 34 clinical strains of S. pyogenes were identified by standard procedures. Antimicrobial susceptibility was analyzed by the Kirby-Bauer method of disk diffusion, and the E-test method was used to determine the MIC to penicillin and erythromycin. All the strains were sensitive to penicillin, clindamycin and vancomycin on disk diffusion. Ten (29.4%) strains were resistant to erythromycin. The pattern of macrolide resistance observed was M type. By the E-test method, 7 (20.6%) strains were penicillin nonsusceptible and 6 (17.6%) were erythromycin resistant. We concluded that surveillance of its susceptibility pattern is crucial to monitoring the development of antibiotic resistance in S. pyogenes.