DETERMINING THE EFFICACY OF ISOXYL, A MYCOLIC ACID INHIBITOR, IN VITRO AGAINST MYCOBACTERIA OTHER THAN MYCOBACTERIUM TUBERCULOSIS (MOTT) STRAINS

Mycobacteria other than Mycobacterium tuberculosis (MOTT) cause infections more commonly in the presence of predisposing factors and underlying diseases.They are also notably resistant to commonly used antituberculosis drugs. Total 11 clinical isolates MOTT were included in the study.Drug susceptibility testing of these isolates was performed by Resistant Ratio method. Minimum inhibitory concentration (MIC) pattern of these isolates of MOTT to mycelia acid synthesis inhibitors namely, Isoxyl(ISO) and Isoniazid (INH) were determined by agar dilution and broth dilution method. Minimum bactericidal concentration (MBC) pattern of these isolates to ISO and INH werealso determined. Out of 11 MOTT isolates, 3 isolates were characterized as Mycobacteriumscrofulaceum, 3 isolates as Mycobacteriumfortuitum, 2 isolates as Mycobacteriumflavescens, 1 isolates as Mycobacterium terrae and 2 isolates as Mycobacteriumkansasi depending upon the results of biochemical tests.The MBC range of INH was found to be 0.025 to 6.4 μg/ml and of ISO was found to be 0.6 to 20 μg/ml. Bactericidal activity of ISO was 7.25 times lower than the activity of INH. It is well known that most MOTT species are more resistant to chemotherapeutic agents other than tubercle bacilli.The inhibitory activity of ISO was more to MOTT strains than Mycobacterium tuberculosis strains.There was low bactericidal activity of ISO to MOTT strains, but better than for Mycobacterium tuberculosis strains.&nbsp

Evaluation of the antimicrobial activity of Gooseberry and Wild Apple fruit juices of the northeast region of India

A large number of wild fruits belonging to the North Eastern region of India are considered to be nutritionally abundant and traditionally used for the treatment of various ailments. In the present study, juices of Gooseberry and Wild Apple have been evaluated for their antimicrobial activity in its consumable form as a potential source of natural anti-infective agent. The fruit juices were screened for their antibacterial and antifungal activities qualitatively, using the agar well diffusion method followed by quantitative assessment by determining the Minimum Inhibitory Concentration (MIC) and Minimum Bactericidal Concentration (MBC). The study revealed that both the fruit juices exhibited broad spectrum antibacterial activity. Considerable activity against drug resistant pathogens such as Methicillin Resistant Staphylococcus aureus (MRSA), Vancomycin Resistant Enterococci (VRE) and Extended Spectrum β-Lactamases (ESBL) producing Gram Negative bacteria (GNB) was observed. Thus, this study highlights the antibacterial efficacy of Wild Apple and Gooseberry fruit juices and augments the beneficiary properties of highly nutritional fruit juices that are specific to the Indian region

TO STUDY INCIDENCE OF MULTI DRUG RESISTANT TUBERCULOSIS IN MUMBAI

Ionized and Rifampicin, the two most potent anti-tuberculosis drugs are rendered ineffective in Multidrug-resistant Tuberculosis (MDR-TB). India, China and Russia contribute to more than 62% of MDR-TB globally. In India, endemic areas like Mumbai are “hotspots” for the dissemination of MDR-TB. The aim of the study was to investigate the incidence of MDR-TB in cases of pulmonary tuberculosis in Mumbai. Total hundred and two clinical isolates of.Tuberculosiswas tested in the study. Drug susceptibility testing of these strains were carried by Resistance Ratio method to ant tuberculosis drugs namely Ionized, Streptomycin andEthambutol and by absolute concentration method for Rifampicinand Pyrazinamide. In our study highest resistance (46 %) was observed to INH followed by RF (42.16 %), SM (29.41 %) and EMB (25.49 %). While, resistance to Pyrazinamide (PZ) was least (7.8%). MDR TB cases were found to be 41.18%.There was significant difference between resistance pattern of INH and EMB, INH and SM, PZ and EMB, PZ and SM, EMB and RF, PZ and RF. (chi square with Yates correction =8.5, p<0.01).Detection of MDR TB strain would not only eliminate non-essential use of antibiotics, but would also help in the selection of most effective drug regimen and guide therapy in chronic cases

Safety and Evaluation of the Immune Response of Coronavirus Nosode (BiosimCovex) in Healthy Volunteers: A Preliminary Study Extending the Homeopathic Pathogenetic Trial

Objectives: Regulatory clinical Phase I studies are aimed at establishing the human safety of an active pharmaceutical agent to be later marketed as a drug. Since homeopathic medicines are prepared by a potentizing method using alcohol, past a certain dilution, their toxicity/infectivity is assumed to be unlikely. We aimed to develop a bridge study between homeopathic pathogenetic trials and clinical trials. The primary purpose was to evaluate the safety of a nosode, developed from clinical samples of a COVID-19 patient. The secondary objectives were to explore whether a nosode developed for a specific clinical purpose, such as use during an epidemic, may elicit laboratory signals worthy of further exploration. Methods: An open-label study was designed to evaluate the safety and immune response of the Coronavirus nosode BiosimCovex, given orally on three consecutive days to ten healthy volunteers. Clinical examinations, laboratory safety and immune parameters were established. Interferon–gamma, Interleukin-6, and CD 4 were measured. (CTRI registration number: CTRI/2020/05/025496). Results: No serious/fatal adverse events were reported. Laboratory tests to measure safety were unchanged. Three subjects showed elevated Interleukin-6 (IL-6) on day 17 in comparison to the baseline, and ten subjects showed elevated IL-6 on day 34. A significant difference between IL-6 observations, calculated by repeated measures ANOVA, was found to be highly significant. On day 60, the IL-6 values of nine subjects were found to return to normal. Corresponding CD4 cell elevation was observed on day 60, when compared to day 34. Conclusions: HPT may potentially extend into physiological changes with regards to immune response and should encourage future studies

PREVALANCE OF CANDIDIASIS IN CHILDREN IN MUMBAI

This study aims to determine the prevalence of infection of Candida species and its resistance to anti-fungal agents amongst children in Mumbai. Total 169 clinical isolates of Candida species from various clinical specimens namely blood, urine, stool; Cerebral spinal fluid, liver abscess,Endotracheal secretion and ventric tip were included in the study. They were screened and identified by staining, culture and microscopy, followed by isolation on specialized chromogenic agar. Antifungal susceptibility tests of these isolates were carried out against Amphotericin B (50mcg), Itraconazole (10mcg), Clotrimazole (10mcg), Ketoconazole (30mcg), Miconazole (30mcg), and Fluconazole (10mcg) and were reported based on CLSI guidelines (2008).The prevalence of Candida infections was found to be greater in males (69%) as compared to female (31%) children. In the age group of 0-5 years, the occurrence of Candida infection was substantially higher as compared to the others age group. Candida albicans was found to be the major causative agent of fungal infections. The susceptibility of isolates to antifungal agents was found to differ in male and female patients. Overall, highest resistance was observed to Fluconazole (27.1%) while, lowest resistance to Amphotericin B (28.7%). Males exhibited highest resistance to Fluconazole (27.1%) while, females showed highest resistance to Amphotericin B (28.7%). Higher resistance was found to the azole group of antifungal agents. Hence, Amphotericin B can be the choice of treatment of Candida infection in children followed by Miconazole.&nbsp

COMPARISON OF CONVENTIONAL METHOD, RADIOMETRIC METHOD AND METHOD USING OXIDATION-REDUCTION DYE FOR DETECTION OF MULTI DRUG RESISTANT TUBERCULOSIS

Today Tuberculosis (TB) has become the most important communicable disease in the world. The emergence of multidrug resistant (MDR) TB has become the main threat to TB treatment and control programs. Rapid detection of TB is critical for the effective treatment of patients. Recently, a method using the Oxidation - Reduction dye has been proposed for drug susceptibility testing of Mycobacterium tuberculosis (M.tuberculosis). In the present study, we have compared Drug Susceptibility Testing (DST) by conventional method, Radiometricmethod like BACTEC460TB (B460TB) System and Oxidation-Reduction Assay such as Microplate Alamar Blue Assay (MABA) for detection of MDR TB. Total 60 clinical isolates of M.tuberculosis were included in the study. The results obtained by MABA for DST of M.tuberculosis were compared with those obtained by B460TB system and Conventional method. DST results obtained by MABA showed good agreement with results obtained by B460TBsystem and Conventional method for Isoniazid and Rifampicin. In our study specificity and sensitivity for susceptible and resistant strain was found to be 98% and 100% for Rifampicin respectively and 100% and 95% for Isoniazid respectively for MABA.MABA appears to be a reliable method for the rapid and simultaneous detection of MDR-TB and DST of M. tuberculosis. It is simple and inexpensive method for DST of M.Tuberculosis with minimal biohazard risk.&nbsp

RELATIONSHIP BETWEEN RPOB MUTATIONS AND MINIMUM INHIBITORY CONCENTRATIONS OF RIFAMPICIN IN MULTI DRUG RESISTANT STRAINS OF MYCOBACTERIUM TUBERCULOSIS

Tuberculosis is an infectious disease caused by obligate intracellular bacteria Mycobacterium tuberculosis, which has created a health problem worldwide. Incidence of Multi-drug resistance tuberculosis creates the scenario dangerous, which is resistant to both the first line drugs namely Rifampicin and ionized. Mutation in rpoB and inhA confers resistance to rifampicin and ionized respectively. In the present study we studied the relationship between rpoB mutations and minimum inhibitory concentrations of Rifampicin in Multi drug resistant(MDR) strains of Mycobacterium tuberculosis. Total 20 MDR strains were selected for the study. The mutations in the rpoB gene were identified by sequencing the PCR products. The minimum inhibitory concentrations of Rifampicin, was determined by Absolute Concentration method. Mutations conferring resistance to Rifampicin in Mycobacterium tuberculosis occurs in the defined region of 81 base pair of rpoB gene. The most frequent mutation found in Rifampicin resistant strain is at position 532, 526 and 516. Mutation occurring at this position results in high level of Rifampicin resistance. Mutation at 511, 518 and 522 is associated with low level of Rifampicin resistance.&nbsp

DETERMINATION OF EFFICACY OF ISOXYL, A MYCOLIC ACID INHIBITOR, IN VITRO AGAINST MYCOBACTERIUM.TUBERCULOSIS STRAINS

The enzymes involved in the biosynthetic pathways of the critical components including mycolic acids offer attractive and selective targets for the developments of novel anti-mycobacterial agents. Isoxyl (ISO), a mycolic acid inhibitor, is an old drug, which was used for the treatment of tuberculosis was evaluated. Determination of Minimum Inhibitory Concentration (MIC) pattern of clinical isolates of Mycobacterium tuberculosis (M. tuberculosis) to mycolic acid inhibitors namely ISO, Isoniazid (INH) and Ethionamide (ETH) by agar and broth dilution Method was done. Also the Minimum bactericidal concentrations were evaluated. Total 40 MDR and 20 susceptible strains of M tuberculosis were tested. The result of the MIC studies showed that ISO is capable of inhibiting the growth of M. tuberculosis in a range of 1-20μg/ml. Inhibitory activity of ISO was higher than activity of ETH in solid media. Amongst three antituberculosis drugs, INH showed highest bactericidal activity against M. tuberculosis strains followed by ETH. While ISO exhibited lowest bactericidal activity. Amongst, three drugs tested, ISO shows highest MBC/MIC ratio with lowest bactericidal activity. ISO showed significantly lower bactericidal activity against MDR strains than susceptible strains of M. tuberculosis. MBC/ MIC ratio of ISO was similar to MDR and susceptible strains of M. tuberculosis. Overall study implies that ISO may be suitable for the treatment of Tuberculosis, particularly multi-drug resistant kind

EVALUATION OF RADIORESPIROMETRIC TECHNIQUE FOR DETECTION OF MYCOBACTERIUM TUBERCULOSIS STRAINS FROM PULMONARY TUBERCULOSIS PATIENTS

A simple and rapid radiorespirometric method, for early detection of tubercle bacilli from sputum samples has been developed. A biphasic liquid scintillation vial system is used for detection of 14CO2 produced by metabolism of acetate 1-2-l4C on glycerol free Lowenstein- Jensen Media (LJM). Total 207 sputum samples were tested by smear, visual growth method (VGM) and radiorespirometric method (RRM) for detection of Mycobacterium tuberculosis (M. tuberculosis) strains. Total 27.54 % sputum samples were scored positive by smear, 21.7% samples by VGM and 33.3 % samples by RRM. There is significant difference between three methods related to sensitivity, specificity, positive and negative predictive value for detection of M. tuberculosis strains. RRM was found to be most sensitive and specific method amongst three. The average detection time by RRM was 9.48 days, compared to 34.47 days by VGM. There is a correlation of number of bacilli in smear and rate of growth in RRM and VGM

Detection of Macrolide, Lincosamide and Streptogramin Resistance among Methicillin Resistant Staphylococcus aureus (MRSA) in Mumbai

Background: The increase in incidence of Methicillin Resistant Staphyloccocus aureus (MRSA) and its extraordinary potential to develop antimicrobial resistance has highlighted the need for better agents to treat such infections. This has led to a renewed interest in use of new drugs for treatment with clindamycin and quinuprsitin-dalfopristin being the preferred choice for treatment. Aim & Objectives: This study was undertaken to detect the prevalence of MacrolideLincosamide-Streptogramin (MLS) resistance among clinical isolates of MRSA.Material and Methods:Two hundred and thirty clinical isolates of S. aureus were subjected to routine antibiotic susceptibility testing including cefoxitin, erythromycin and quinupristindalfopristin. Inducible resistance to clindamycin was tested by 'D' test as per Clinical and Laboratory Standards Institute (CLSI) guidelines. Results: Out of all S. aureus isolates, 93.91% were identified as MRSA. In the disc diffusion testing, 81.5% of isolates showed erythromycin resistance. Among these, the prevalence of constitutive (cMLS ), inducible (iMLS ) b b and MS-phenotype were 35.80%, 31.82% and 32.39% respectively by the D-test method. 77.8% of isolates were resistant to quinupristin-dalfopristin and the Minimum Inhibitory Concentration (MIC) ranged from 4–32 µg/ml. 89.20% of isolates were resistant to both quinupristin-dalfopristin and erythromycin of which 35.03%, 35.67% and 29.30% belonged to iMLS , cMLS and MS phenotype respectively. Conclusion: The emergence of quinupristindalfopristin resistance and MLS phenotypes brings b about the need for the simple and reliable D-test in routine diagnosis and further susceptibility testing for proper antimicrobial therapy