Interaction effects of entomopathogenic nematodes and insecticides for the management of grubs of Holotrichia longipennis and Brahmina coriacea

Aim: Laboratory studies were conducted to study the potential of entomopathogenic nematodes and their interaction with the commonly used insecticides viz., clothianidin, imidacloprid and chlorpyriphos against first, second and third instar grubs of H. longipennis and B. coriacea.Methodology: Two entomopathogenic nematodes viz., Heterorhabditis indica Poinar and Steinernema caprocapsae Weiser were tested against I-III instar grubs of Brahmina coriacea (Hope) and Holotrichia longipennis (Blanchard) under laboratory conditions @1600IJs per cup by soil application. The mortality data was recorded after every 24 hrs up to 5 days. For evaluating the interaction effects among different tested biocontrol agents with insecticides, each entomopathogenic nematode species and insecticides was tested alone and in combination. H. indica and S. carpocapsae were applied @ 400IJs per cup. Insecticides were applied at a lower concentrations viz. 150, 100 and 75 ppm for chlorpyriphos, imidacloprid and clothianidin, respectively and were applied on the same day of nematode application with a gap of 2 hrs. Mortality data was taken after 24 hrs. Results: Individual application of H. indica and S. carpocapsae @1600 IJs per cup had clearly lethal effect and produced mortality to the tune of 55.5-90.7 % in both the species of white grubs. H. indica revealed higher virulence as compared to S. carpocapsae against both the species. Combination of both the species with all the tested insecticides yielded synergistic interactions and H. indica+ imidacloprid combination proved to be the most promising among all the tested combinations. Interpretation: Tested entomopathogens and insecticide combination produced synergistic interactions more effectively to first, second and early third instar grubs (H. longipennis and B. coriacea).

Diagnostic challenges posed by a rare alpha globin chain variant Hb Fontainebleau in a pregnant female and its potential effects in her children in view of multiple globin gene defects in her husband

Alpha globin chain variants per se do not cause severe morbidity and mortality but can modify – usually ameliorate – the clinical manifestations of beta globin chain variants when co-inherited with the latter. They also pose challenges in interpretation of high-performance liquid chromatography histograms and require molecular analysis for proper characterization. Hemoglobin (Hb) Fontainebleau is a rare alpha globin chain variant [alpha 21(B2) Ala?Pro], of which only three families have been reported from India in the past. Here, we describe a case of Hb fontainebleau detected in heterozygous condition in a 19-year-old primigravida. Her husband was found to have a double heterozygous state for HbQ India and beta-thalassemia trait. This opens up the possibility of multiple combinations of hemoglobinopathies in the offspring.

Conflict of interest disclosure and interpretation – rest assured the medical professional in the audience is perceptive, alert and smart

INTRODUCTION:We present data from a systematic survey on conflict of interest (COI) disclosure and its interpretation by the doctors participating in continuing medical education (CME).METHODS:A brief 12 question online Google survey with multiple choice options (read, select, and click) was done among Indian practicing doctors using links shared through WhatsApp through the internet over a 72 h period.RESULTS:Of the 386 replies, 373 unique replies were eligible for evaluation. The majority found CME activities beneficial. About 73% of participants would watch out for bias, even if the speaker shows COI disclosure slide. The use of brand/trade names was considered as a flag for bias by the majority. About 99% wanted the speaker to show a final take home message slide. Cross verification of the data presented by comparing to published data was done in more than 75% of instances by only 25% of the participating doctors. A significantly higher number of doctors found bias when CME activities were being organized by the health-care industry as compared to programs of medical bodies/societies/organizations.DISCUSSION:COI considerations are given due to the importance of medical professionals. However, doctors are smart enough to understand the limitations of such disclosures and remain alert to ensure they are not influenced by any bias. Take home message slide gives the presenters opportunity to share their insights and allows the audience to make their own judgment on the impartiality of the data presented. The doctors are aware that bias could be more when CME activities are organized by healthcare industry and take appropriate precautions.CONCLUSION:COI is is given due importance by the medical professionals. COI disclosures are often incomplete. Doctors remain alert to ensure they are not influenced by biased presentations. Concluding take home message slide is unanimously recommended. Presentation bias is more when healthcare industry is directly organizing educational and promotional activities.

Our patient at risk from unauthorized, unapproved, illegally imported fake “generic medicines”

Introduction: We came across patients inquiring with oncologists about cheap copy medicines. These were of recently licensedinnovator drugs that should have been available from original company holding their patents. In fact these copy medicines weremanufactured in our neighboring countries and made available in India. We investigated further and this manuscript puts together thestartling information that we were able to find regarding the thriving grey market for fake generic medicines.

Antiretroviral treatment, viral load of mothers & perinatal HIV transmission in Mumbai, India.

Background & objectives: Mother-to-child transmission (MTCT) is the most significant route of HIV transmission in children below the age of 15 yr. In India, perinatal HIV transmission, even after treatment, accounts for 5.4 per cent of HIV cases. The present study was conducted to evaluate the efficacy of anti-retro viral therapy (ART) or prophylactic treatment (PT) to control maternal viral load in HIV positive women, and its effect on vertical HIV transmission to their infants. Methods: A total of 58 HIV positive women were enrolled at the time of delivery and their plasma samples were obtained within 24 h of delivery for estimation of viral load. Viral load analysis was completed in 38 women. Infants received single dose nevirapine within 2 h of birth and zidovudine for 6 wk. At the end of 18 month follow up, HIV positive or negative status was available in 28 infants. Results: Results revealed undetectable levels of viral load in 58.3 per cent of women with ART compared to 30.7 per cent of women with PT. No women on ART had viral load more than 10,000 copies/ml, whereas seven (26.9%, P=0.07) women receiving PT had this viral load. Median CD4 count of women on PT (483 cells/μl) was high compared to the women on ART (289 cells/ μl). At the end of 18 months follow up, only two children were HIV positive, whose mothers were on PT. One had in utero transmission; infection detected within 48 h of delivery, while the other child was infected post partum as HIV was detected at six months follow up. Interpretation & conclusions: Women who received a single dose of nevirapine during delivery had higher levels of viral load than women on ART. Combination drug therapy for pregnant women is now a standard of care in most of the western countries; use of nevirapine monotherapy at the time of delivery in our settings is not effective in controlling viral load. This highlights initiation of ART in pregnant women to control their viral load and thus to inhibit mother to child HIV transmission.

Polymorphisms in major cytokine genes: A study among human immunodeficiency Virus-1 serodiscordant couples in Mumbai, India.

Purpose: Polymorphism in cytokine genes may affect its production, which play an important role in modulation of human immunodeficiency virus (HIV) infection. Evaluation of these polymorphisms might help to understand why some individuals remain uninfected in spite of several exposures to HIV infection, such as the negative spouses of discordant couples. The aim of this study was to evaluate the association of 22 single nucleotide polymorphisms (SNPs) in 13 cytokine genes and their receptors with HIV infection in serodiscordant couples, attending the Integrated Counselling and Testing Centre of a Municipality Hospital. Materials and Methods: At the end of at least 2 years of follow up, 42 couples were confirmed as being serodiscordant. Genotyping was carried out in blood samples of these couples using the polymerase chain reaction-sequence-specific amplification method. Results: Significantly high frequency of interleukin-1 receptor antagonist IL-1RA mspa 11100 CC ( P = 0.04), tumor necrosis factor-alpha TNF-α −238 AG ( P = 0.01) and IL-4 −33 TT ( P = 0.01) was observed in HIV seropositives (HSP) while frequency of TNF-α −238 GG ( P = 0.02) was significantly high among the exposed uninfected (EU). However, application of Bonferroni correction identified only two SNPs i.e., TNF-α −238 AG and IL-4 −33 TT to be significantly associated with the acquisition of HIV. In remaining cytokine genes, no significant association was observed. Conclusion: Our study highlighted possible association of certain specific polymorphisms with HIV transmission, whereas presence or absence of certain other polymorphism in EU individuals might be offering protection from HIV infection. These variations at the genetic level might help to explore new insights into treatment and HIV prevention strategies.

Evaluation of Calypte AWARE HIV-1/2 OMT antibody test as a screening test in an Indian setting.

Purpose: Integrated counselling and testing centres (ICTC) provide counselling and blood testing facilities for HIV diagnosis. Oral fluid tests provide an alternative for people whodo not want blood to be drawn. Also, it avoids the risk of occupational exposure. The goal of this study was to evaluate the utility of Calypte AWARE HIV-1/2 OMT antibody test as a screening test in an Indian setting. Materials and Methods: A cross-sectional study was carried out after ethics committee approval in 250 adult ICTC clients. Blood was collected and tested from these clients for HIV diagnosis as per routine policy and the results were considered as the gold standard. Also, after another written informed consent, oral fluid was collected from the clients and tested for the presence of HIV antibodies. Twenty five clients who had and 25 clients who had not completed their secondary school education (Group A and Group B, respectively) were also asked to perform and interpret the test on their own and their findings and experiences were noted. Result: The sensitivity, specificity, PPV and NPV of the oral fluid antibody test were 100%, 98.51%, 94.11% and 100%, respectively. Seventy six percent of clients preferred oral fluid testing. Group B found it difficult to perform the test as compared to Group A and this difference was statistically significant (P ≤ 0.05). Conclusion: Oral fluid testing can be used as a screening test for HIV diagnosis; however, confirmation of reactive results by blood-based tests is a must.

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.

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.

Enterococcal infections with special reference to phenotypic characterization & drug resistance.

BACKGROUND & OBJECTIVES: Enterococci, classified as group D streptococci, are the second leading cause of nosocomial infections. The incidence of enterococcal infections and species prevalent in India is not thoroughly investigated. The present study was undertaken to isolate and characterize enterococci from clinical specimens and determine the antimicrobial susceptibility pattern of these isolates. METHODS: Clinical specimens (blood, urine and swabs) were cultured on bile esculin azide agar (BEAA) for isolation of enterococci. The phenotype based scheme included Gram staining of growth on BEAA and subculturing of cocci on sheep blood agar plates for vancomycin disk diffusion and hydrolysis of L-pyrrolidonyl-beta-napthylamide (PYR) testing. The phenotypic method was used to surveillance cultures that yielded growth on BEAA. Enterococcal strains were further identified to the species level by conventional biochemical tests. PYR positive isolates were further characterized into vancomycin resistant enterococci (VRE) and nonVRE depending upon vancomycin inhibition zone size. The isolates were characterized into vanA, vanB and vanC depending upon minimum inhibitory concentration (MIC) values. Conventional method was used to study the antibiogram of isolates. RESULTS: A total of 52 isolates of enterococci (10 Enterococcus faecalis, 42 E. faecium) were isolated from 534 clinical specimens. Of the 52 isolates, 12 isolates were resistant to vancomycin with an MIC > 4 microg/ml but sensitive to teicoplanin (vanB isolates). INTERPRETATION & CONCLUSION: Our study reveals the problem of multiple drug resistant enterococci and emergence of VRE. Better susceptibility tests need to be used to measure the vancomycin resistance accurately.

An evaluation of routine immunization coverage in some districts of West Bengal and Assam.

A coverage evaluation survey was carried out in the five districts of West Bengal and Goalpara district of Assam during the period from November 2002 to April 2003 through 40 clusters sampling technique. Results revealed that highest coverage with routine immunization was in Paschim Medinipur (82.5%) followed by Kolkata (71.6%), Malda (65.3%), 24 Parganas South (61.9%) districts of West Bengal. Murshidabad district of West Bengal had only 41.3% coverage while poorest coverage was observed in Goalpara district (27.2%) of Assam. "Not aware of the need for routine immunization" was the main reason for not being vaccinated with all the UIP vaccines. Vaccination was given mainly through government hospitals in Kolkata while it was administered mainly through subcentres in the other rural districts.

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.

Persistent strongyloidiasis in a case of common variable immunodeficiency.

A case of strongyloidiasis in common variable immunodeficiency is reported here. The patient was given several courses of albendazole without any response.

Diet nutrition and health profile of elderly population of urban Baroda.

Geriatric population forms a significant proportion of our total population. Hence, various problems affecting the overall health of the elderly need special consideration. In this context, studies were undertaken to assess the socio-demographic factors, diet and health profile of 320 elderly men and women of all the three income groups of Urban Baroda. Data on socio-demographic factors was collected using an open ended questionnaire. Nutritional status was assessed using anthropometric measurements of height, weight, mid upper arm circumference (MUAC) and body mass index (BMI). Information on dietary profile was collected by 24 hour dietary recall method. Fasting practices were also studied. Socio-demographic data of geriatric men of high, middle and low income groups revealed that majority of the subjects were married. A greater percentage of high income group (HIG) men had nuclear family whereas majority of low income groups (LIG) elderly men resided in a joint family. Socio-demographic profile of elderly women of all the 3 income groups revealed that most of the subjects were Hindus. The percentage of widowhood, illiteracy and joint family system were higher in LIG as compared to the elderly women in middle and high income groups (MIG and HIG). Nutrient intake data of elderly men of all the income groups revealed lower consumption of energy, protein, iron and beta-carotene as compared to the RDA whereas fats and vitamin C intakes were higher as compared to the RDA (p < 0.05). The mean nutrient intake, by the LIG elderly women, in terms of energy, protein, iron, calcium, beta-carotene and vitamin C were significantly (p < 0.05) lower than the RDA as well as when compared to the elderly women of MIG and HIG. Mean anthropometric measurements of weight and BMI were higher in elderly HIG and MIG men as compared to the elderly men from LIG. Significant difference was found in all the anthropometric measurements of elderly women of LIG as compared to HIG and MIG. Morbidity profile showed a striking rise in problems of oral cavity, cardio vascular disease, neurological problems and problems of gastro intestinal tract with advancing age in both elderly men and women. The study reveals striking differences in diet, health and disease profile with advancing age.

Seroprevalence of IgG antibodies to Helicobacter pylori among gastrointestinal endoscopists, radiologists and paramedical personnel.

BACKGROUND: The route of transmission of Helicobacter pylori is unknown. Since the organism has been isolated from saliva, gastric juice and stool, medical personnel could be at high risk for acquiring the infection during procedures like gastrointestinal endoscopy. AIMS: To study whether endoscopy is a professional hazard for acquisition of H. pylori. METHODS: We studied the prevalence of IgG antibodies to H. pylori in endoscopists (n = 17), radiologists (n = 17) and personnel from paraclinical branches (n = 35); microbiology (n = 21), pathology (n = 7) and forensic medicine (n = 7); among the paraclinical personnel five were at high risk because they worked with cultures of H. pylori. Subjects answered a questionnaire regarding upper gastrointestinal symptoms, and precautions taken at the work place against infection. The serum was tested for IgG antibodies to H. pylori using a microwell ELISA and a rapid card test. RESULTS: H. pylori antibodies were present in five (29.4%) endoscopists, three (17.6%) radiologists and seven (20%) paraclinical personnel; only one of the 5 high risk para medical personnel was positive. There was no correlation between the duration of performing endoscopies and the H. pylori IgG status. CONCLUSION: Endoscopy is not a risk factor for acquiring H. pylori infection.

Dental plaque: an unlikely reservoir of Helicobacter pylori.

BACKGROUND AND AIM: Dental plaque has been suggested as a reservoir for Helicobacter pylori, though data in this regard are conflicting. We evaluated the prevalence of H. pylori DNA in dental plaque using polymerase chain reaction (PCR). METHODS: Antral H. pylori status of 156 patients with acid-peptic disease (APD) was studied by rapid urease test (RUT), histology and culture. Dental plaque obtained from these 156 patients and 92 healthy volunteers was evaluated for the presence of H. pylori using RUT, culture and PCR. RESULTS: H. pylori was present in 133 antral biopsy samples by RUT and/or histology. The dental plaque of 37 patients with APD and 21 healthy volunteers tested positive by RUT. H. pylori was not isolated by culture from any of the dental plaques. PCR gave a significant amplification product in 11 of 248 (4.4%) dental plaque samples, 7 from patients with APD and 4 from normal healthy volunteers. CONCLUSION: The frequency of H. pylori in the dental plaque is low, and this is unlikely to be a prominent site of infection with H. pylori.