Effect of different methods of pasteurization on bactericidal action of human milk: A prospective observational study

Background & objectives: Pasteurization involves not only inactivation of pathogens, but also loss of immunological functions and bactericidal action of human milk. Hence, this study was aimed to explore the stability of such bactericidal action after subjecting human milk samples to thermal pasteurization under different condition of time and temperature. Methods: In this observational study 48 human milk samples were analyzed over a period of three months. The effect of holder and flash methods of pasteurization on bactericidal action against Escherichia coli was evaluated compared to the control sample before and after 72 h of storage at ?18癈. Results: Both holder and flash methods of pasteurization showed significant reduction in the E. coli growth to 46.4 and 25.5 per cent, respectively, after 24 h of incubation (P <0.001). The bactericidal activity was significantly more in samples subjected to holder method compared to flash method before and after 72 h of storage (46.41�.38 vs. 25.50�.74, P <0.001 and 42.27�.38 vs. 18.33�.55, P <0.001). Interpretation & conclusions: Our results showed that the bactericidal activity of human milk was better preserved by the holder method of pasteurization. Further well-powered and well-designed randomized trials are needed to confirm the findings.

Improved case detection using Xpert Mycobacterium tuberculosis/rifampicin assay in skeletal tuberculosis

Background: In India, musculoskeletal tuberculosis (TB) accounts for 10%�% of extrapulmonary TB. Data on drug-resistant skeletal TB are lacking. At present, the diagnosis is based mainly on radiological techniques. Laboratory confirmation of skeletal TB is delayed as 6�weeks are required for culture results. Xpert Mycobacterium tuberculosis/rifampicin (MTB/RIF) assay is a fully automated test which simultaneously detects MTB and RIF resistance within 3 h. Hence, this study was done to compare the yield of case detection using Xpert assay in comparison with culture in specimens received from clinically suspected skeletal TB cases. Methods: Retrospective analysis of microscopy, culture and Xpert assay results was carried out on specimens received in laboratory from skeletal TB cases from January 2016 to December 2017. Results: Of the 201 patients analysed, majority of the specimens were obtained from the spine (55.72%). MTB was detected in 48.68% of tissue and 24% of pus specimens. Xpert assay was detected MTB in 67 (33.33%) specimens of which 53 (47.32%) were from the spine. Culture was detected MTB in 66 (32.83%) specimens. Xpert assay was detected two specimens more than culture. One specimen was positive by only culture. RIF-resistant MTB was detected in 10 (14.92%) specimens by Xpert assay. Conclusion: The spine is the most common site involved. Tissue specimen is better for early diagnosis. High RIF resistance in skeletal TB is an alarming situation. Ability of Xpert MTB/RIF assay for rapid and simultaneous detection of MTB and RIF resistance in comparison with culture makes it a useful diagnostic tool in skeletal TB.

Study of HIV-1 subtypes in serodiscordant couples attending an integrated counselling and testing centre in Mumbai using heteroduplex mobility analysis and DNA sequencing.

Aims: To determine the prevalent subtypes of HIV-1 in serodiscordant couples. Setting: Integrated Counselling and Testing Centre (ICTC), Department of Microbiology. Study Design: Prospective pilot study. Participants: Thirty HIV-1 serodiscordant couples. Inclusion Criteria: a) Documentation of HIV-1 infection in one partner and seronegative status in the other, current history of continued unprotected sexual activity within the partnership, demonstration that they have been in a partnership for at least 1 year and are not currently on highly active antiretroviral therapy HAART; b) willingness of both partners to provide written informed consent including consent to continued couple counselling for 3 months. Materials and Methods: HIV-1 subtyping was carried out by heteroduplex mobility analysis (HMA) by amplifying env region; and DNA sequencing by amplifying gag region. Results: HIV-1 env gene was amplified successfully in 10/30 samples; gag gene, in 25/30 samples; and both env and gag gene were amplified successfully in 5/30 samples. HIV-1 subtype C was detected from 21 samples; subtype B, from 7; and subtype A, from 2. Sample from 1 positive partner was detected as subtype C by env HMA and subtype B by gag sequencing. Conclusion: HIV-1 subtype C was found to be the predominant subtype of HIV-1 in serodiscordant couples attending our ICTC, followed by HIV-1 subtype B and HIV-1 subtype A, respectively. DNA sequencing was found to be the most reliable method for determining the subtypes of HIV-1.

A rare case of tubercular cerebellar abscess.

Tubercular brain abscess are uncommon and tubercular cerebellar abscess are rarely reported. Most of these cases occur in immunocompromised patients. We report a case of multiple cerebellar abscesses in a 55-year-old HIV seronegative non-diabetic female, who complained of headache, neck pain and unsteadiness of gait since two months. She had been on treatment for pulmonary tuberculosis, diagnosed earlier. Diagnosis was made by CT scan of brain and confirmed by bacteriological examination of drained pus obtained by suboccipital craniotomy. The patient showed signs of recovery.

Rapid antibiotic susceptibility testing of Mycobacterium tuberculosis: its utility in resource poor settings.

PURPOSE: To compare the rapid colorimetric nitrate reductase based antibiotic susceptibility (CONRAS) test performed on Mycobacterium tuberculosis isolates with the conventional method i.e, the proportion method. METHODS: One hundred clinical isolates of M. tuberculosis were tested for susceptibility to isoniazid (INH) and rifampicin (RIF) by the conventional proportion method and CONRAS in Middlebrook 7H9 liquid medium enriched with growth supplements (MB7H9S). RESULTS: The performance of the CONRAS test was evaluated using proportion method as the gold standard. The sensitivity (ability to detect true drug resistance) and specificity (ability to detect true drug susceptibility) of the CONRAS test to INH was 93.75 and 98.52% and for RIF it was 96.10 and 100% respectively. The mean time for reporting was 6.3 days and the test showed excellent reproducibility. The kappa (k) value for INH was 0.92 and for RIF was 0.99, indicating excellent agreement between the two methods. CONCLUSIONS: CONRAS test is a rapid and reliable method of drug susceptibility for M. tuberculosis.

Immunological response to two hepatitis B vaccines administered in two different schedules.

OBJECTIVES: To evaluate the antibody response to a plasma derived hepatitis B vaccine containing pre-S and S antigens and a recombinant vaccine containing only S antigen and compare the two vaccines in 0, 1, 2 and 0, 1, 6, schedules METHODS: One hundred fifty nine healthy infants were randomized to receive the vaccines in two different schedules. Anti HBs titers were estimated prior to the vaccination and then one-month following the second and third doses. RESULTS: Both the vaccines produced a high rate of seroconversion and seroprotection. The antibody response was similar with both the vaccines and the 0, 1, 6, schedule induced a significantly higher antibody level than the 0, 1, 2, schedule. CONCLUSION: 0, 1, 6 schedule produces a higher antibody level than 0, 1, 2 schedule though both the schedules have good seroconversion and seroprotection rates with both the vaccines.

Correlation of fine needle aspiration cytology, smear and culture in tuberculous lymphadenitis: a prospective study.

BACKGROUND AND AIM: Bacteriological studies are necessary to confirm the diagnosis of tuberculous lymphadenitis, as cytological appearances mimic other granulomatous lesions. The objective was to assess the diagnostic role of culture of fine needle aspiration done on clinically suspected cases of tuberculous lymphadenitis and to determine the prevalence of drug resistance in M. tuberculosis isolates. SETTING AND DESIGN: A prospective, double-blind study over a period of one year in a tertiary care hospital. MATERIAL AND METHODS: Fine needle aspiration cytology and culture were done on 250 patients with clinical suspicion of tuberculous lymphadenitis. STATISTICAL ANALYSIS: Data was statistically analysed using chi square test. Sensitivity, specificity, positive predictive and negative predictive values and likelihood ratio were also calculated. RESULT: Of the 161 cytologically or microbiologically proven cases of tuberculous lymphadenitis, cytological changes consistent with tuberculosis were observed in 133 patients, out of which mycobacteria were isolated in 102 aspirates. Mycobacteria were also isolated from 28 aspirates cytologically missed as tuberculous lymphadenitis. Of the 130-mycobacterial isolates, 5 were non-tuberculous mycobacteria. Culture positivity was significantly higher (P<0.001) than smear positivity. Drug susceptibility studies showed resistance to one or more drugs in 61% of isolated strains with maximum resistance to isoniazid (16% primary and 48% secondary) and minimum to ethambutol (4% primary and 12% secondary). CONCLUSION: Culture for mycobacteria should be carried out on all aspirates from patients suspected with tuberculous lymphadenitis.