Socio-demographic and behavioural determinants of HIV prevalence among homosexual and bisexual men having sex with men (MSM) in India: Integrated bio-behavioural surveillance.

Background: HIV prevalence among men who have sex with men (MSM) is 16.5 times higher than adult HIV prevalence. With a socio-cultural context that demands marriage, a considerable proportion of MSM in India have female sexual partners and act as a bridge population. Stratified analysis of HIV risk factors among homosexual and bisexual MSM will be instrumental in identifying the high-risk MSM. We aim to identify the socio-demographic and behavioural factors associated with HIV risk among homosexual and bisexual MSM. Methods: Overall, 23081 MSM were enrolled in the IBBS conducted across India between October 2014 and November 2015. Data and blood samples were collected. Chi-square test, univariate and multivariable logistic regression methods were used in data analysis. Results: HIV prevalence was significantly higher among homosexual MSM than bisexual MSM. Older age, lesser education, being a sex worker, being married, living with a male or hijra partner, younger age at initiation of MSM behaviour, duration, injecting drugs, and having STI symptoms were associated with higher prevalence. The prevalence of new homosexual MSM was 11.4%. Nearly 75% of the bisexual MSM reported inconsistent condom usage with female partners. Conclusion: Interventions for early identification of new MSM and advocacy for safe sex with alternative preventive techniques are recommended.

Spatial heterogeneity of risk factors associated with HIV prevalence among men who inject drugs in India: An analysis of the data from the integrated bio-behavioral surveillance, India.

People who inject drugs (PWID) are India's third-largest vulnerable population to human immunodeficiency virus (HIV) infection. PWID in India are confined to certain geographic locations and exhibit varying injecting and sexual risk behaviors, contributing considerably to increasing HIV trends in specific regions. Spatial heterogeneity in risk factors among vulnerable PWID influences HIV prevalence, transmission dynamics, and disease management. Stratified analysis of HIV prevalence based on risk behaviors and geographic locations of PWID will be instrumental in strategic interventions. To stratify the male PWID based on their risk behaviors in each state and determine the HIV prevalence for each stratum. The behavioral data and HIV prevalence of the national integrated biological and behavioural surveillance (IBBS), a nationwide cross-sectional community-based study conducted in 2014 to 2015, was analyzed. Data from 19,902 men who inject drugs across 53 domains in 29 states of India were included. Women who inject drugs were excluded at the time of IBBS, and hence PWID in this study refers to only men who inject drugs. PWID were categorized based on their risk profile, and the corresponding HIV prevalence for each state was determined. HIV prevalence was the highest (29.6%) in Uttar Pradesh, with a high prevalence of risk behaviors among PWID. High HIV prevalence ranging between 12.1% and 22.4% was observed in a few states in East and North-East India and most states in central and North India. Unsafe injecting and sexual practices were significantly (P < .05) associated with higher HIV prevalence and more significantly in National Capital Territory of Delhi (P < .001). Unsafe injecting practices among PWID were proportionally higher in Western and Central India, whereas unsafe sexual behaviors were widespread among most states. Unsafe sexual practices among male PWID were common. The high prevalence of unsafe injecting had significant HIV infection and transmission risks in Western and Central India. The results emphasize the need for stratified, region-specific interventions and combination approaches for harm reduction among PWID. Strengthening the measures that facilitate the reduction of high-risk behaviors, adoption of safe practices, and utilization of HIV services will positively impact HIV prevention measures among PWID.

Injecting drug use & HIV prevalence among female sex workers: Evidence from the National Integrated Biological & Behavioural Surveillance, India.

Background & objectives: Female sex workers (FSWs) who inject drugs (FSW-IDs) have a higher risk of HIV infection and transmission. Understanding the socio-demographic characteristics and other risk behaviours among FSW-IDs will help in strengthening targeted interventions for HIV prevention and management. In the present study, the HIV prevalence, associated socio-demographic characteristics and risk behaviours among FSWs who injected drugs (FSW-IDs) and those who did not ID (FSW-NIDs) was determined in India. Methods: The national cross-sectional, community-based, integrated biological and behavioural surveillance was conducted in 2014-2015 at 73 randomly selected FSW domains across 28 States and Union Territories in India. The sample size was fixed at 400 for each domain, and a probability-based sampling method was followed. The data were analyzed by logistic regression methods. Results: Data from 27,007 FSWs were included in the analysis, of which 802 (3%) were FSW-IDs. HIV prevalence among FSW-IDs was significantly higher than that in FSW-NIDs (4.5 vs. 1.9%). Univariate analysis showed that factors significantly associated with higher HIV prevalence among FSW-IDs were older age, sex work as the only source of income, dissolved marriage, living with a sex worker, urban locality of sex work and consumption of alcohol or oral drugs. In multivariable analysis, factors such as older age of FSW-IDs (35 yr and above), having a dissolved marriage and sex work being the only source of income were observed to be independently and significantly associated with higher HIV prevalence. Interpretation & conclusions: Scaling up the HIV preventive interventions for FSW-IDs, such as facilitating awareness and improved access to needle and syringe exchange programme (NSEP) and opioid substitution therapy (OST), encouraging safe sex and injecting practices, educating on the harmful effects of alcohol and drugs and providing alternative vocation options to secure their financial needs are several strategies that may reduce HIV transmission among FSWs.

Sub-regional Trend of HIV Infection Among Antenatal Clinic Attendees in Andhra Pradesh (2003-2019).

INTRODUCTION: The analyses of local risks at sub-national levels and tailored interventions are being emphasized for effective HIV management. Andhra Pradesh (AP) is a high HIV prevalence state in India, with the second-highest number of People with HIV (PWH) and a consistent decline in HIV prevalence at the state level. Probing further into the region and district-wise levels and trends of HIV prevalence will provide critical insights into sub-regional epidemic patterns. Hence, this paper analyzes the regional and district-level trends of HIV prevalence among pregnant women attending antenatal clinics (ANC) from 2003 to 2019 in AP, South India. METHODS: HIV prevalence data collected from pregnant women in AP during HIV Sentinel Surveillance (HSS) between 2003 and 2019 was used for trend analysis. The consistent sites were grouped into three regions (Coastal Andhra, Rayalaseema and Uttar Andhra), totaling 39 sites, including 21 rural and 18 urban sites. Regional and district-level HIV prevalence was analyzed using the Chisquare trend test, and spatial analysis was done using QGIS software. For the last three HSS rounds, HIV prevalence based on sociodemographic variables was calculated to understand the factors contributing to HIV positivity in each region. RESULTS: In total, 143,211 pregnant women were recruited. HIV prevalence in AP was 0.30% (95% CI: 0.22 - 0.39) in 2019. The prevalence was 0.31%, 0.35% and 0.22% in Coastal Andhra, Rayalaseema and Uttar Andhra, respectively. HIV prevalence had significantly (P < 0.05) declined in all regions. The overall trend indicated that the HIV prevalence was higher among older pregnant women and in urban regions. However, recent trends consistently report HIV positivity among young and new pregnant mothers, illiterates, and rural regions. CONCLUSION: The overall trend indicated that the HIV prevalence was higher among older pregnant women and in urban regions. However, recent trends consistently report HIV positivity among young and new pregnant mothers, illiterates, and rural regions. Identifying the contextual risk patterns associated with HIV transmission will further improve HIV preventive and management programs among the general population.

An Analysis of Levels and Trends in HIV Prevalence Among Pregnant Women Attending Antenatal Clinics in Karnataka, South India, 2003-2019.

BACKGROUND AND OBJECTIVE: Periodic tracking of the trends and the levels of HIV prevalence at regional and district levels helps to strengthen a state's HIV/AIDS response. HIV prevalence among pregnant women is crucial for the HIV prevalence estimation of the general population. Karnataka is one of the high HIV prevalence states in India. Probing regional and district levels and trends of HIV prevalence provides critical insights into district-level epidemic patterns. This paper analyzes the region- and district-wise levels and trends of HIV prevalence among pregnant women attending the antenatal clinics (ANC) from 2003 to 2019 in Karnataka, South India. METHODS: HIV prevalence data collected from pregnant women in Karnataka during HIV Sentinel Surveillance (HSS) between 2003 and 2019 was used for trend analysis. The consistent sites were grouped into four zones (Bangalore, Belgaum, Gulbarga and Mysore regions), totaling 60 sites, including 30 urban and 30 rural sites. Regional and district-level HIV prevalence was calculated; trend analysis using Chi-square trend test and spatial analysis using QGIS software was done. For the last three HSS rounds, HIV prevalence based on sociodemographic variables was calculated to understand the factors contributing to HIV positivity in each region. RESULTS: In total, 254,563 pregnant women were recruited. HIV prevalence in Karnataka was 0.22 (OR: 0.15 95% CI: 0.16 - 0.28) in 2019. The prevalence was 0.24, 0.32, 0.17 and 0.14 in Bangalore, Belgaum, Gulbarga, and Mysore regions, respectively. HIV prevalence had significantly (P< 0.05) declined in 26 districts. CONCLUSION AND GLOBAL HEALTH IMPLICATIONS: HIV prevalence among pregnant women was comparatively higher in Bangalore and Belgaum regions. Analysis of contextual factors associated with the transmission risk and evidence-based targeted interventions will strengthen HIV management in Karnataka. Regionalized, disaggregated, sub-national analyses will help identify emerging pockets of infections, concentrated epidemic zones and contextual factors driving the disease transmission.

Hospital-based sentinel surveillance for bacterial meningitis in under-five children prior to the introduction of the PCV13 in India.

INTRODUCTION: A hospital-based sentinel surveillance network for bacterial meningitis was established in India to estimate the burden of bacterial meningitis, and the proportion of major vaccine-preventable causative organisms. This report summarises the findings of the surveillance conducted between March 2012, and September 2016 in eleven hospitals. METHODS: We enrolled eligible children with bacterial meningitis in the age group of one to 59 months. CSF samples were collected and processed for biochemistry, culture, latex agglutination, and real-time PCR. Pneumococcal isolates were serotyped and tested for antimicrobial susceptibility. RESULTS: Among 12 941 enrolled suspected meningitis cases, 586 (4.5%) were laboratory confirmed. S. pneumoniae (74.2%) was the most commonly detected pathogen, followed by H. influenzae (22.2%), and N. meningitidis (3.6%). Overall 58.1% of confirmed bacterial meningitis cases were children aged between one, and 11 months. H. influenzae meningitis cases had a high (12.3%) case fatality rate. The serotypes covered in PCV13 caused 72% pneumococcal infections, and the most common serotypes were 14 (18.3%), 6B (12.7%) and 19F (9.9%). Non-susceptibility to penicillin was 57%. Forty-five (43.7%) isolates exhibited multidrug resistance, of which 37 were PCV13 serotype isolates. CONCLUSIONS: The results are representative of the burden of bacterial meningitis among under-five children in India. The findings were useful in rolling out PCV in the National Immunization Program. The non-susceptibility to penicillin and multidrug resistance was an important observation. Timely expansion of PCV across India will significantly reduce the burden of antimicrobial resistance. Continued surveillance is needed to understand the trend after PCV expansion in India.

Heterogeneity and Confinement of HIV Prevalence among Pregnant Women Calls for Decentralized HIV Interventions: Analysis of Data from Three Rounds of HIV Sentinel Surveillance in Karnataka: 2013-2017.

BACKGROUND: The HIV sentinel surveillance (HSS) serves to estimate the levels and trend of HIV prevalence among high-risk, bridge, and general population and monitors HIV management at national, state, and district levels. Data from HSS are valuable in understanding the risk factors associated with HIV transmission with particular demographic characteristics. OBJECTIVES: The objective was to analyze the sociodemographic profile of the pregnant mothers attending the antenatal care (ANC) clinics in Karnataka, in order to understand the dynamics of HIV within the general population in Karnataka. MATERIALS AND METHODS: Study design: this was a cross-sectional study conducted using consecutive sampling method. Study setting: the surveillance was conducted at select antenatal clinics, in Karnataka, India, between January and March, in the years 2013, 2015, and 2017. Methodology: in total, 74,278 eligible pregnant women aged between 15 and 49 years, attending the sentinel sites for the first time during the surveillance period, were included in the study. Information on their sociodemographic characteristics and blood samples was collected. RESULTS: HIV prevalence among the ANC clinic attendees has significantly declined, reaching a recent stabilization. The risk factors significantly associated with HIV among pregnant women were age, education, occupation, and marital status. HIV is highly concentrated in the northern and southern districts of Karnataka. CONCLUSION: Despite the declining trends of HIV prevalence in Karnataka, the epidemic is heterogeneous and concentrated within the state, calling for decentralized region-specific interventions.

Prevalence and Risk Factors Associated with HIV Infection among Pregnant Women in Odisha State, India.

BACKGROUND AND OBJECTIVES: The purpose of this study was to analyze trends in HIV prevalence and risk factors associated with HIV infection among pregnant women attending antenatal clinics in Odisha State, India. METHODS: Data were from the HIV Sentinel Surveillance (HSS) among pregnant women, a descriptive cross-sectional study using consecutive sampling method and conducted in India. Data and samples were collected from pregnant women attending select antenatal clinics that act as designated sentinel sites in Odisha State, India, during the three months surveillance period and in three surveillance years: 2013, 2015, and 2017. All eligible pregnant women aged between 15 and 49 years, attending the sentinel sites for the first time during the surveillance period, were included. Information on their socio-demographic characteristics and blood samples were also collected. RESULTS: In total, 38,384 eligible pregnant women were included in the survey. Of these, 107 women were HIV positive, with an overall prevalence of 0.28%. HIV prevalence indicated a stabilizing trend between 2013 and 2017. However, pregnant women whose spouses were non-agricultural laborers, truck drivers, or migrants were significantly at higher risk of being infected. Likewise, HIV prevalence significantly increased over the years among pregnant women whose spouses were in the service sector (government or private). District-wise fluctuations in HIV prevalence was observed, with the district of Cuttack recording the highest prevalence among the districts. CONCLUSION AND GLOBAL HEALTH IMPLICATIONS: Women who are spouses of non-agricultural laborers, truck drivers or migrants need focused interventions, such as creating awareness on HIV and its prevention. Migration, due to poverty and its impact on sexually transmitted diseases among migrants from low and middle-income countries, have been documented globally. Single male migrant specific interventions are recommended to halt the disease progression among pregnant women and general population in Odisha, India.

Substance use and risk of HIV infection among Men who have Sex with Men in India: Analysis of National IBBS data, India.

HIV prevalence is higher among Men who have Sex with Men (MSM), owing to their unsafe sexual behavior. Further, MSM indulge in behaviors such as consumption of alcohol/oral drugs and/or injecting during/before sex that poses the risk of unsafe behaviors, thereby increasing their vulnerability to HIV. The study aims to analyze the factors associated with HIV infection among the multi-risk MSM using any substances with those MSM who do not use substances.Community-based cross-sectional survey design using probability-based sampling between October 2014 and November 2015.For the nation-wide Integrated Biological and Behavioral Surveillance (IBBS), 23,081 MSM were recruited from 4067 hotspots in 108 districts across India. Information on demographics, sexual behaviors, substance use, sexual partners, and awareness on HIV and its management was collected from the consented respondents using computer-assisted personal interview (CAPI) by trained personnel. Blood samples were tested for HIV. Statistical analyses were done, to study the associations between substance use and its influence on high-risk sexual behaviors and HIV infection.One in 3 MSM (33.88%) in India were substance users, thus exhibiting "multi-risk" (MR) behaviors. Significantly higher HIV prevalence (3.8%, P < .05) was reported among MR-MSM, despite 97.2% of them being aware of HIV. Higher HIV prevalence among MSM exhibiting homosexual behavior for ≤1 year is of specific concern, as this accounts to recent infections and indicates the increased vulnerability of the infection among the new entrants.Substance-use resulting in high-risk sexual behavior was significantly associated with higher HIV prevalence among MR-MSM. Integrated targeted interventions focusing on safe sex and safe-IDU practices among MR-MSM are required to end the disease transmission.

The relationship between educational attainment and hiv prevalence among pregnant women attending antenatal clinics in six states of India: Sentinel surveillance from 2010 to 2017.

BACKGROUND: The National AIDS Control Programme provides support for HIV prevention services to pregnant women attending antenatal care (ANC) clinics through testing, kit delivery, counseling, and treatment services. The impact of HIV prevention programs in the general population is assessed by monitoring trends and progress made against the HIV epidemic among pregnant women attending ANC clinics during HIV Sentinel Surveillance (HSS). OBJECTIVES: This study explores the association of HIV risk with educational attainment for Indian women across different age groups from four repeated cross-sectional surveillance of antenatal clinics in six states from the southern part of India. METHODS: Data collected from the repeated cross-sectional HSS conducted during the year 2010-2011 (baseline) and 2016-2017 (end line) across six states were used for this analysis. The total sample size was 94,266 at baseline and 99,434 at end line. In the logistic regression analysis, we focused on identifying the association between educational attainment, and HIV prevalence adjusting for period effects across two age groups for women attending ANC clinics. RESULTS: The analysis showed an inverse association between education and HIV risk across different age groups. The age-segregated and survey period adjusted analysist showed that for older women (≥25 years), the HIV risk in 2010 ranged from 41% lower among 5th Grade to 80% lower among postgraduates than illiterates. For the <25 year age group, this risk of HIV for pregnant women was 35% to 49% lower. CONCLUSIONS: To ensure an effective national response to control and prevent HIV infection, policymakers in India need to focus on ≥25 years' age group of women attending ANC for designing educational interventions to reduce HIV risk as well as the prevention of mother-to-child transmission of HIV.

Losartan Rescues Inflammation-related Mucociliary Dysfunction in Relevant Models of Cystic Fibrosis.

Rationale: Despite therapeutic progress in treating cystic fibrosis (CF) airway disease, airway inflammation with associated mucociliary dysfunction remains largely unaddressed. Inflammation reduces the activity of apically expressed large-conductance Ca2+-activated and voltage-dependent K+ (BK) channels, critical for mucociliary function in the absence of CFTR (CF transmembrane conductance regulator).Objectives: To test losartan as an antiinflammatory therapy in CF using CF human bronchial epithelial cells and an ovine model of CF-like airway disease.Methods: Losartan's antiinflammatory effectiveness to rescue BK activity and thus mucociliary function was tested in vitro using primary, fully redifferentiated human airway epithelial cells homozygous for F508del and in vivo using a previously validated, now expanded pharmacologic sheep model of CF-like, inflammation-associated mucociliary dysfunction.Measurements and Main Results: Nasal scrapings from patients with CF showed that neutrophilic inflammation correlated with reduced expression of LRRC26 (leucine rich repeat containing 26), the γ subunit mandatory for BK function in the airways. TGF-ß1 (transforming growth factor ß1), downstream of neutrophil elastase, decreased mucociliary parameters in vitro. These were rescued by losartan at concentrations achieved by nebulization in the airway and oral application in the bloodstream: BK dysfunction recovered acutely and over time (the latter via an increase in LRRC26 expression), ciliary beat frequency and airway surface liquid volume improved, and mucus hyperconcentration and cellular inflammation decreased. These effects did not depend on angiotensin receptor blockade. Expanding on a validated and published nongenetic, CF-like sheep model, ewes inhaled CFTRinh172 and neutrophil elastase for 3 days, which resulted in prolonged tracheal mucus velocity reduction, mucus hyperconcentration, and increased TGF-ß1. Nebulized losartan rescued both mucus transport and mucus hyperconcentration and reduced TGF-ß1.Conclusions: Losartan effectively reversed CF- and inflammation-associated mucociliary dysfunction, independent of its angiotensin receptor blockade.

Nipah virus epidemic in southern India and emphasizing "One Health" approach to ensure global health security.

Nipah virus (NiV) encephalitis first reported in "Sungai Nipah" in Malaysia in 1999 has emerged as a global public health threat in the Southeast Asia region. From 1998 to 2018, more than 630 cases of NiV human infections were reported. NiV is transmitted by zoonotic (from bats to humans, or from bats to pigs, and then to humans) as well as human-to-human routes. Deforestation and urbanization of some areas have contributed to greater overlap between human and bat habitats resulting in NiV outbreaks. Common symptoms of NiV infection in humans are similar to that of influenza such as fever and muscle pain and in some cases, the inflammation of the brain occurs leading to encephalitis. The recent epidemic in May 2018 in Kerala for the first time has killed over 17 people in 7 days with high case fatality and highlighted the importance of One Health approach. The diagnosis is often not suspected at the time of presentation and creates challenges in outbreak detection, timely control measures, and outbreak response activities. Currently, there are no drugs or vaccines specific for NiV infection although this is a priority disease on the World Health Organization's agenda. Antivirals (Ribavirin, HR2-based fusion inhibitor), biologicals (convalescent plasma, monoclonal antibodies), immunomodulators, and intensive supportive care are the mainstay to treat severe respiratory and neurologic complications. There is a great need for strengthening animal health surveillance system, using a One Health approach, to detect new cases and provide early warning for veterinary and human public health authorities.

Diabetic retinopathy and its risk factors in patients with type 2 diabetes attending rural primary healthcare facilities in Tamil Nadu.

BACKGROUND: India has a high burden of diabetic retinopathy ranging from 12.2% to 20.4% among patients with type 2 diabetes mellitus (T2DM). A T2DM management programme was initiated in the public sector in Tamil Nadu. We estimated the prevalence of diabetic retinopathy and its associated risk factors. METHODS: We did a cross-sectional survey among patients with T2DM attending two primary health centres for treatment and follow-up in Kancheepuram, Tamil Nadu in January- March 2013. We did a questionnaire-based survey, and measured blood pressure and biochemical parameters (serum creatinine, plasma glucose, etc.) of the patients. We examined their eyes by direct and indirect ophthalmoscopy and defined diabetic retinopathy using a modified classification by Klein et al. We calculated the proportion and 95% CI for the prevalence and adjusted odds ratio (AOR) for risk factors associated with diabetic retinopathy. RESULTS: Among the 270 patients, the mean (SD) age was 54.5 (10) years. The median duration of T2DM was 48 months. The prevalence of diabetic retinopathy was 29.6%. Overall, 65.9% of patients had hypertension, 14.4% had nephropathy (eGFR <60 mg/dl) and 67.4% had neuropathy. Among patients with comorbid conditions, 60%, 48%, 32%, and 3% were already diagnosed to have hypertension, neuropathy, retinopathy, and nephropathy, respectively. The risk factors for diabetic retinopathy were hypertension (AOR 3.2, 95% CI 1.7-6.3), duration of T2DM >5 years (AOR 6.5, 95% CI 3.6-11.7), poor glycaemic control (AOR 2.4, 95% CI 1.4-4.4), and nephropathy (AOR 2.3, 95% CI 1.1-4.6). CONCLUSIONS: There was a high burden of undetected retinopathy and other comorbid conditions among patients with T2DM. Early detection of comorbid conditions and glycaemic control can be improved by training care-providers and educating patients.

Cholera outbreak linked with lack of safe water supply following a tropical cyclone in Pondicherry, India, 2012.

In the aftermath of a severe cyclonic storm on 7 January 2012, a cluster of acute diarrhoea cases was reported from two localities in Pondicherry, Southern India. We investigated the outbreak to identify causes and recommend control measures. We defined a case as occurrence of diarrhoea of more than three loose stools per day with or without vomiting in a resident of affected areas during 6-18 January 2012. We used active (door-to-door survey) and stimulated passive (healthy facility-based) surveillance to identify cases. We described the outbreak by time, place, and person. We compared the case-patients with up to three controls without any apparent signs and symptoms of diarrhoea and matched for age, gender, and neighbourhood. We calculated matched odds ratio (MOR), 95% confidence intervals (CI), and population attributable fractions (PAF). We collected rectal swabs and water samples for laboratory diagnosis and tested water samples for microbiological quality. We identified 921 cases and one death among 8,367 residents (attack rate: 11%, case-fatality: 0.1%). The attack rate was the highest among persons of 50 years and above (14%) and females (12%). The outbreak started on 6 January and peaked on the 9th and lasted till 14 January. Cases were clustered around two major leakages in water supply system. Nine of the 16 stool samples yielded V. cholerae O1 Ogawa. We identified that consumption of water from the public distribution system (MOR=37, 95% CI 4.9-285, PAF: 97%), drinking unboiled water (MOR=35, 95% CI 4.5-269, PAF: 97%), and a common latrine used by two or more households (MOR=2.7, 95% CI 1.3-5.6) were independently associated with cholera. Epidemiological evidence suggested that this outbreak was due to ingestion of water contaminated by drainage following rains during cyclone. We recommended repair of the water supply lines, cleaning-up of the drains, handwashing, and drinking of boiled water.

Chloroquine efficacy studies confirm drug susceptibility of Plasmodium vivax in Chennai, India.

BACKGROUND: Assessing the Plasmodium vivax burden in India is complicated by the potential threat of an emerging chloroquine (CQ) resistant parasite population from neighbouring countries in Southeast Asia. Chennai, the capital of Tamil Nadu and an urban setting for P. vivax in southern India, was selected as a sentinel site for investigating CQ efficacy and sensitivity in vivax malaria. METHODS: CQ efficacy was evaluated with a 28-day in vivo therapeutic study, while CQ sensitivity was measured with an in vitro drug susceptibility assay. In both studies, isolates also underwent molecular genotyping to investigate correlations between parasite diversity and drug susceptibility to CQ. Molecular genotyping included sequencing a 604 base pair (bp) fragment of the P. vivax multidrug resistant gene-1 (Pvmdr1) for single nucleotide polymorphisms (SNPs) and also the amplification of eight microsatellite (MS) loci located across the genome on eight different chromosomes. RESULTS: In the 28-day in vivo study (N=125), all subjects were aparasitaemic by Day 14. Passive case surveillance continuing beyond Day 28 in 22 subjects exposed 17 recurrent infections, which ranged from 44 to 148 days post-enrollment. Pvmdr1 sequencing of these recurrent infections revealed that 93.3% had identical mutant haplotypes (958M/Y976/1076L) to their baseline Day 0 infection. MS genotyping further revealed that nine infection pairs were related with ≥ 75% haplotype similarity (same allele at six or more loci). To test the impact of this mutation on CQ efficacy, an in vitro drug assay (N=68) was performed. No correlation between IC50 values and the percentage of ring-stage parasites prior to culture was observed (r(sadj): -0.00063, p = 0.3307) and the distribution of alleles among the Pvmdr1 SNPs and MS haplotypes showed no significant associations with IC50 values. CONCLUSIONS: Plasmodium vivax was found to be susceptible to CQ drug treatment in both the in vivo therapeutic drug study and the in vitro drug assay. Though the mutant 1076 L of Pvmdr1 was found in a majority of isolates tested, this single mutation did not associate with CQ resistance. MS haplotypes revealed strong heterogeneity in this population, indicating a low probability of reinfection with highly related haplotypes.

Comparison of ITS and IGS1 regions for strain typing of clinical and non-clinical isolates of Pichia anomala.

Pichia anomala is an emerging nosocomial pathogen and there is a need for methods that distinguish between different P. anomala strains. In the typing of several clinical as well as non-clinical P. anomala strains, the sequence variation of the internal transcribed spacer (ITS) was found to be inadequate for typing purposes. The intergenic spacer 1 (IGS1) region of the rDNA of several P. anomala strains was therefore investigated in detail. The IGS1 region (which varied from 1213 to 1231 bp in length) was interspersed with repeats and had more variation than the ITS regions. Comparative analysis in cases where analysis by the ITS was ambiguous clearly revealed the IGS1 region to be a more discriminatory tool in the typing of P. anomala strains.