Perceived needs of disease vector control programs: A review and synthesis of (sub)national assessments from South Asia and the Middle East.

Systems for disease vector control should be effective, efficient, and flexible to be able to tackle contemporary challenges and threats in the control and elimination of vector-borne diseases. As a priority activity towards the strengthening of vector control systems, it has been advocated that countries conduct a vector-control needs assessment. A review was carried out of the perceived needs for disease vector control programs among eleven countries and subnational states in South Asia and the Middle East. In each country or state, independent teams conducted vector control needs assessment with engagement of stakeholders. Important weaknesses were described for malaria, dengue and leishmaniases regarding vector surveillance, insecticide susceptibility testing, monitoring and evaluation of operations, entomological capacity and laboratory infrastructure. In addition, community mobilization and intersectoral collaboration showed important gaps. Countries and states expressed concern about insecticide resistance that could reduce the continued effectiveness of interventions, which demands improved monitoring. Moreover, attainment of disease elimination necessitates enhanced vector surveillance. Vector control needs assessment provided a useful planning tool for systematic strengthening of vector control systems. A limitation in conducting the vector control needs assessment was that it is time- and resource-intensive. To increase the feasibility and utility of national assessments, an abridged version of the guidance should focus on operationally relevant topics of the assessment. Similar reviews are needed in other regions with different contextual conditions.

Bamboo stumps that are artificially in use put pressure on Dengue and Chikungunya vector control in Dhaka City, Bangladesh.

BACKGROUND OBJECTIVES: Aedes aegypti and Ae. albopictus mosquitoes breed in natural and artificial containers, and they transmit dengue and chikungunya. Therefore, a study was conducted to identify the contribution of bamboo stumps to these disease vectors that were used in the flower garden as pillars to hold the bamboo flex fence. METHODS: Two sizes of whole bamboo were used to hold fences around gardens at Dhaka University, Bangladesh, and they were painted red and green. Mosquito larvae and pupae were collected from bamboo stumps between July and August, and vectors were identified up to the species level. The data were analyzed using the STATA/MP 14.2 version. RESULTS: We found 83.5% and 0.2% were Ae. albopictus and Ae. aegypti, respectively, and the rest were Culex and Armigeres species. Ae. albopictus, Ae. aegypti, and both species-positive bamboo stumps were 46.9, 0.7, and 47.1 percent, respectively. 54.5% of bamboo stumps had at least one mosquito species. The average stump depth for Aedes positive stumps (mean =11.7 cm, SE = 0.5) was significantly (p <0.001) higher than the Aedes negative stumps (mean = 9.5 cm, SE = 0.4). 53.8% and 38.0% stumps were found Aedes positive on the ground and upper sides of fences, respectively, and found significant (p<0.01) differences between both sides. A zero-inflated negative binomial count model is significant at a 5% level of significance, χ2(4) = 11.8, p = 0.019 (<0.05) for Ae. albopictus. Stump depth is found to have a significant positive effect on the number of Aedes-positive stumps. INTERPRETATION CONCLUSION: Artificially used natural containers are adding pressure to current mosquito control activities as mosquitoes are breeding on them, which needs additional attention.

Effect of dual sintering with laser irradiation and thermal treatment on printed copper nanoparticle patterns.

The dual sintering of copper (Cu) nanoparticles (NPs) was introduced to produce conductive patterns suitable for flexible electronics applications. In this method, laser irradiation using a Nd:YAG laser with a wavelength of 1064 nm was performed at laser powers of 400, 600 and 800 mJ. The laser irradiation time was 15 and 30 s for each laser power. After laser irradiation, all of the Cu NP patterns were thermally sintered under formic acid vapors. The temperature and time for thermal treatment were selected as 260 °C and 15 min, respectively. The resultant physical, chemical, electrical and mechanical properties were evaluated and compared considering the six different dual sintering conditions. The Cu NP patterns sintered using 800 mJ for 30 s showed increased necking and coalescence compared to the other patterns and featured a microstructure with increased density. Despite being oxidized, the Cu NP patterns sintered with 800 mJ for 30 s showed the lowest electrical resistivity of 11.25µΩ cm. The surface of every sintered Cu pattern was oxidized, and mechanical hardness increased with increasing laser power. The Cu NP pattern sintered with 800 mJ for 30 s demonstrated the highest hardness of 48.64 N mm-2. After sintering using the six different conditions, the Cu NP patterns exhibited a weight loss of 0.02-3.87 wt%, and their roughness varied in the range of 26.15-74.08 nm. This can be attributed to the effective removal of organic residues and the degree of particle agglomeration. After performing folding tests up to 50 cycles, Cu NP patterns showed an upward trend in resistance with increasing laser power and time. The highest and lowest resistance ratios were observed as 3.97 and 17.24 for the patterns sintered at 400 mJ for 15 s and 800 mJ for 30 s, respectively.

Assessment of Knowledge, Attitudes, and Practices toward Kala-azar in an Endemic District in Bangladesh: A Cross-Sectional Survey.

We undertook a study to assess current knowledge, attitudes, and practices related to kala-azar to advise the national kala-azar elimination program in Bangladesh. A community-based cross-sectional study was conducted in two endemic subdistricts (upazilas): Fulbaria and Trishal. Based on upazila health complex surveillance data, one endemic village was selected randomly from each of these subdistricts. A total of 511 households (HHs) (261 in Fulbaria and 250 in Trishal) were included in the study. An adult from each HH was interviewed using a structured questionnaire. Specifically, data were collected on knowledge, attitudes, and practices related to kala-azar. Of the respondents, 52.64% were illiterate. All study participants had heard approximately kala-azar, and 30.14% of the HHs or neighboring HHs have had at least one kala-azar case. Of the respondents, 68.88% knew that kala-azar is transmitted through sick people, and more than 56.53% of the study participants said that mosquitoes transmitted kala-azar, even though 90.80% were aware of the presence of sand flies. Of the participants, 46.55% were aware that insect vectors laid their eggs in the water. The Upazila Health Complex was the preferable health-care facility for 88.14% of the villagers. In addition, 62.03% used bed nets for preventing sand fly bites and 96.48% of the families had mosquito nets. Based on these observations, the national program should strengthen its current community engagement activities to increase the knowledge of kala-azar in endemic communities.

Prevalence of epilepsy in Bangladesh: Results from a national household survey.

OBJECTIVE: To determine the prevalence and types of epilepsy in Bangladesh. METHODS: We conducted a nationwide population-based cross-sectional survey among Bangladeshi population of all ages, except children under one month. We surveyed 9839 participants (urban, 4918; rural, 4920) recruited at their households using multistage cluster sampling. Trained physicians with neurology background confirmed the diagnosis of suspected epilepsy cases identified by interviewer-administered questionnaires. We reported the overall and sex, residence, and age groups-specific prevalence of epilepsy per 1000 populations with 95% confidence interval. RESULTS: The national prevalence of epilepsy per 1000 was 8.4 (95% CI 5.6-11.1), urban 8.0 (4.6-11.4), and rural 8.5 (5.60-11.5). The prevalence in adult males and females was 9.2 (5.7-12.6) and 7.7 (3.6-11.7), respectively. The prevalence in children aged <18 years (8.2, 3.4-13.0 was similar to adults (8.5 (5.4-11.4). Among all epilepsy cases, 65.1% had active epilepsy. Their (active epilepsy) prevalence was 5.8 (3.5-8.1). Of them, 63.4% were not receiving treatment. Moreover, those who received allopathy treatment, 72.5% had low adherence leading to a high treatment gap. SIGNIFICANCE: Our findings out of this first-ever national survey were similar to other Asian countries. However, the prevalence of active epilepsy and treatment gap were considerably higher. This study serves useful evidence for tailoring interventions aimed to reduce the burden of epilepsy-primarily through targeted community awareness program-and access to antiepileptic treatment in health facilities in Bangladesh.

Effect of insecticide-treated bed nets on visceral leishmaniasis incidence in Bangladesh. A retrospective cohort analysis.

BACKGROUND: Visceral leishmaniasis (VL) is a parasitic disease, transmitted by the sand fly species Phlebotomus argentipes in the Indian sub-continent. Effective vector control is highly desirable to reduce vector density and human and vector contact in the endemic communities with the aim to curtail disease transmission. We evaluated the effect of long lasting insecticide treated bed nets (LLIN) and bed nets impregnated with slow-release insecticide tablet K-O TAB 1-2-3 (jointly insecticide-treated nets or ITN) on VL incidence in a highly endemic sub-district (upazila) in Bangladesh. METHODS: Several distributions of LLIN or K-O TAB 1-2-3 for self-impregnation of bed nets at home took place in Fulbaria upazila, Mymensigh district from 2004 to 2008 under three research projects, respectively funded by CDC, Atlanta, USA (2004) and WHO-TDR, Geneva, Switzerland (2006 & 2008). We included all households (n = 8142) in the 20 villages that had benefited in the past from one of these interventions (1295 donated LLIN and 11,918 local bed nets impregnated with K-O TAB 1-2-3) in the "exposed cohort". We recruited a "non-exposed cohort" in villages with contemporaneously similar incidence rates who had not received such vector control interventions (7729 HHs from nine villages). In both cohorts, we visited all families house to house and ascertained any VL cases for the 3 year period before and after the intervention. We evaluated the incidence rate (IR) of VL in both cohorts as primary endpoint, applying the difference-in-differences method. RESULTS: The study identified 1011 VL cases (IR 140.47/10,000 per year [py]) before the intervention, of which 534 and 477 cases in the intervention and control areas respectively. The IR was 144.13/10,000 py (534/37050) and 136.59/10,000 py (477/34923) in the intervention and control areas respectively, with no significant difference (p = 0.3901) before the intervention. After the intervention, a total of 555 cases (IR 77.11/10,000 py) were identified of which 178 (IR 48.04/10,000 py) in the intervention and 377 (107.95/10,000 py) in the control area. The intervention area had a significant lower IR than the control area during follow up, rate difference = -59.91, p<0.0001. The IR during follow up was significantly reduced by 96.09/10,000 py in the intervention area (p<0.0001) and 28.63/10,000 py in control area (p<0.0001) compared to baseline. There was a strong and significant overall effect of the ITN intervention, δ = -67.45, p <0.0001. Sex (OR = 1.36, p<0.0001) and age (OR = 0.99, p<0.0001) also had a significant effect on VL incidence. Male had a higher risk of VL than female and one year increase in age decreased the likelihood of VL by about 0.92%. Two third of the VL incidence occurred in the age range 2 to 30 years (median age of VL patients was 17 years). CONCLUSION: VL incidence rate was significantly lower in the ITN intervention cohort compared to control in Bangladesh. Some bias due to more intense screen-and-treat activities or other interventions in the intervention area cannot be ruled out. Nonetheless, given their feasibility and sustainability, ITNs should be considered for integrated vector control during the maintenance phase of the VL elimination programme.

Role of latent female genital tuberculosis in recurrent early pregnancy loss: A retrospective analysis.

BACKGROUND: Latent Female Genital tuberculosis (FGTB) or tubercular infestation is prevalent in Southeast Asia and even the presence of tubercular bacilli in the genital tract is becoming an important factor for reproductive failure. An immature endometrium becomes non-receptive, preventing implantation or rejection of implanted embryo in early months, resulting in recurrent pregnancy loss (RPL) in association with other factors. OBJECTIVE: To detect the underlying causes of RPL in addition to the proven causes like uterine cavity defects, thrombophilia, chromosomal abnormalities, etc. MATERIALS AND METHODS: 317 women with RPL, enrolled over a period of 60 months (January 2014 to December 2018) conducted at Calcutta Fertility Mission in the present study. They were grouped in A, B, and C and undergone routine tests for the same along with the PCR test with an endometrial aspirate. RESULTS: Patients with only latent FGTB (Group A), patients with FGTB and associated factors (Group B), and patients with other causes of RPL (other than latent FGTB) (Group C) were34.4%, 42.3%, and 23.3% respectively. About 29.36%, 47.01%, and 21.62%of the patients had achieved pregnancy in Group A, B, and C, respectively. The rate of miscarriage was high in both Groups A and B, affected with latent FGTB, and live-birth was higher (75%) in Group C that did not have tubercular involvement of the genital tract. CONCLUSION: The tubercular infestation or latent FGTB as per our study appears to be a very important cause of RPL in patients with recurrent "unexplained" miscarriage. It should be treated adequately at an early stage to prevent permanent damage to pelvic organs and restore reproductive health in women.

The role of case proximity in transmission of visceral leishmaniasis in a highly endemic village in Bangladesh.

BACKGROUND: Visceral leishmaniasis (VL) is characterised by a high degree of spatial clustering at all scales, and this feature remains even with successful control measures. VL is targeted for elimination as a public health problem in the Indian subcontinent by 2020, and incidence has been falling rapidly since 2011. Current control is based on early diagnosis and treatment of clinical cases, and blanket indoor residual spraying of insecticide (IRS) in endemic villages to kill the sandfly vectors. Spatially targeting active case detection and/or IRS to higher risk areas would greatly reduce costs of control, but its effectiveness as a control strategy is unknown. The effectiveness depends on two key unknowns: how quickly transmission risk decreases with distance from a VL case and how much asymptomatically infected individuals contribute to transmission. METHODOLOGY/PRINCIPAL FINDINGS: To estimate these key parameters, a spatiotemporal transmission model for VL was developed and fitted to geo-located epidemiological data on 2494 individuals from a highly endemic village in Mymensingh, Bangladesh. A Bayesian inference framework that could account for the unknown infection times of the VL cases, and missing symptom onset and recovery times, was developed to perform the parameter estimation. The parameter estimates obtained suggest that, in a highly endemic setting, VL risk decreases relatively quickly with distance from a case-halving within 90m-and that VL cases contribute significantly more to transmission than asymptomatic individuals. CONCLUSIONS/SIGNIFICANCE: These results suggest that spatially-targeted interventions may be effective for limiting transmission. However, the extent to which spatial transmission patterns and the asymptomatic contribution vary with VL endemicity and over time is uncertain. In any event, interventions would need to be performed promptly and in a large radius (≥300m) around a new case to reduce transmission risk.

Indoor residual spraying for kala-azar vector control in Bangladesh: A continuing challenge.

BACKGROUND: Visceral leishmaniasis (VL) in the Indian subcontinent is a fatal disease if left untreated. Between 1994 to 2013, the Ministry of Health of Bangladesh reported 1,09,266 cases of VL and 329 VL related deaths in 37 endemic districts. Indoor residual spraying (IRS) using dichlorodiphenyltrichloroethane (DDT) was used by the national programme in the 1960s to control malaria. Despite findings of research trials demonstrating that the synthetic pyrethroid deltamethrin 5 WP was very effective at reducing vector densities, no national VL vector control operations took place in Bangladesh between 1999 to early 2012. In 2012, IRS using deltamethrin 5 WP was re-introduced by the national programme, which consisted of pre-monsoon spraying in eight highly endemic sub-districts (upazilas). The present study aims to evaluate the effectiveness of IRS on VL vectors, as well as the process and performance of the spraying activities by national programme staff. METHODS: Five highly endemic upazilas of Mymensingh district were purposively selected (Fulbaria, Trishal, Mukthagacha, Gaforgaon and Bhaluka) to conduct the present study using the WHO/TDR monitoring and evaluation tool kit. IRS operations, conducted by 136 squads/teams, and 544 spraymen, were observed using check lists and questionnaires included in the WHO/TDR monitoring and evaluation tool kit. A household (HH) acceptability survey of IRS was conducted in all study areas using a structured questionnaire in 600 HHs. To measure the efficacy of IRS, pre-IRS (two weeks prior) and post-IRS (at one and five months after), vector density was measured using CDC light traps for two consecutive nights. Bioassays, using the WHO cone-method, were carried out in 80 HHs (40 sprayed and 40 unsprayed) to measure the effectiveness of the insecticide on sprayed surfaces. RESULTS: Of the 544 spraymen interviewed pre-IRS, 60%, 3% and 37% had received training for one, two and three days respectively. During spraying activities, 64% of the spraying squads had a supervisor in 4 upazilas but only one upazila (Mukthagacha) achieved 100% supervision of squads. Overall, 72.8% of the spraying squads in the study upazilas had informed HHs members to prepare their houses prior to spraying. The required personal protective equipment was not provided by the national programme during our observations and the spraying techniques used by all sprayers were sub-standard compared to the standard procedure mentioned in the M&E toolkit. In the HH interviews, 94.8% of the 600 respondents said that all their living rooms and cattle sheds had been sprayed. Regarding the effectiveness measurements (i.e. reduction of vector densities), a total of 4132 sand flies were trapped in three intervals, of which 3310 (80.1%) were P. argentipes; 46.5% (1540) males and 53.5% (1770) females. At one month post-IRS, P. argentipes densities were reduced by 22.5% but the 5 months post-IRS reduction was only 6.4% for both male and female. The bioassay tests showed a mean corrected mortality of P. argentipes sand flies at one month post-IRS of 87.3% which dropped to 74.5% at 4 months post-IRS in three upazilas, which is below the WHO threshold level (80%). CONCLUSION: The national programme should conduct monitoring and evaluation activities to ensure high quality of IRS operations as a pre-condition for achieving a fast and sustained reduction in vector densities. This will continue to be important during the maintenance phase of VL elimination on the Indian subcontinent. Further research is needed to determine other suitable vector control option(s) when the case numbers are very low.

Susceptibility of field-collected Phlebotomus argentipes (Diptera: Psychodidae) sand flies from Bangladesh and Nepal to different insecticides.

BACKGROUND: The sand fly Phlebotomus argentipes is the vector for visceral leishmaniasis (VL) in the Indian sub-continent. In Bangladesh since 2012, indoor residual spraying (IRS) was applied in VL endemic areas using deltamethrin. In Nepal, IRS was initiated in 1992 for VL vector control using lambda-cyhalothrin. Irrational use of insecticides may lead to vector resistance but very little information on this subject is available in both countries. The objective of this study was to generate information on the susceptibility of the vector sand fly, P. argentipes to insecticide, in support of the VL elimination initiative on the Indian sub-continent. METHODS: Susceptibility tests were performed using WHO test kits following the standard procedures regarding alpha cypermethrin (0.05%), deltamethrin (0.05%), lambda-cyhalothrin (0.05%), permethrin (0.75%), malathion (5%) and bendiocarb (0.1%) in six upazilas (sub-districts) in Bangladesh. In Nepal, the tests were performed for two insecticides: alpha cypermethrin (0.05%) and deltamethrin (0.05%). Adult P. argentipes sand flies were collected in Bangladesh from six VL endemic upazilas (sub-districts) and in Nepal from three endemic districts using manual aspirators. RESULTS: The results show that VL vectors were highly susceptible to all insecticides at 60 minutes of exposure in both countries. In Bangladesh, corrected mortality was 100% at 15 minutes as well as 30 minutes of exposure. The study sites in Nepal, however, showed some diverse results, with a mortality rate less than 90% for 15 minutes of exposure with alpha cypermethrin and deltamethrin in two districts but was above 95% after 30 minutes of exposure. CONCLUSIONS: These results suggest that the insecticides tested can still be used in the national programmes of Bangladesh and Nepal. However, insecticide rotation should be performed to mitigate the possible development of insecticide resistance. Periodic susceptibility tests should be performed by the countries to get timely alerts regarding insecticide resistance.

Control of Phlebotomus argentipes (Diptera: Psychodidae) sand fly in Bangladesh: A cluster randomized controlled trial.

BACKGROUND: A number of studies on visceral leishmaniasis (VL) vector control have been conducted during the past decade, sometimes came to very different conclusion. The present study on a large sample investigated different options which are partially unexplored including: (1) indoor residual spraying (IRS) with alpha cypermethrin 5WP; (2) long lasting insecticide impregnated bed-net (LLIN); (3) impregnation of local bed-nets with slow release insecticide K-O TAB 1-2-3 (KOTAB); (4) insecticide spraying in potential breeding sites outside of house using chlorpyrifos 20EC (OUT) and different combinations of the above. METHODS: The study was a cluster randomized controlled trial where 3089 houses from 11 villages were divided into 10 sections, each section with 6 clusters and each cluster having approximately 50 houses. Based on vector density (males plus females) during baseline survey, the 60 clusters were categorized into 3 groups: (1) high, (2) medium and (3) low. Each group had 20 clusters. From these three groups, 6 clusters (about 300 households) were randomly selected for each type of intervention and control arms. Vector density was measured before and 2, 4, 5, 7, 11, 14, 15, 18 and 22 months after intervention using CDC light traps. The impact of interventions was measured by using the difference-in-differences regression model. RESULTS: A total of 17,434 sand flies were collected at baseline and during the surveys conducted over 9 months following the baseline measurements. At baseline, the average P. argentipes density per household was 10.6 (SD = 11.5) in the control arm and 7.3 (SD = 8.46) to 11.5 (SD = 20.2) in intervention arms. The intervention results presented as the range of percent reductions of sand flies (males plus females) and rate ratios in 9 measurements over 22 months. Among single type interventions, the effect of IRS with 2 rounds of spraying (applied by the research team) ranged from 13% to 75% reduction of P. argentipes density compared to the control arm (rate-ratio [RR] ranged from 0.25 to 0.87). LLINs caused a vector reduction of 9% to 78% (RR, 0.22 to 0.91). KOTAB reduced vectors by 4% to 73% (RR, 0.27 to 0.96). The combination of LLIN and OUT led to a vector reduction of 26% to 86% (RR, 0.14 to 0.74). The reduction for the combination of IRS and OUT was 8% to 88% (RR, 0.12 to 0.92). IRS and LLIN combined resulted in a vector reduction of 13% to 85% (RR, 0.15 to 0.77). The IRS and KOTAB combination reduced vector densities by 16% to 86% (RR, 0.14 to 0.84). Some intermediate measurements for KOTAB alone and for IRS plus LLIN; and IRS plus KOTAB were not statistically significant. The bioassays on sprayed surfaces or netting materials showed favourable results (>80% mortality) for 22 months (IRS tested for 12 months). In the KOTAB, a gradual decline was observed after 6 months. CONCLUSIONS: LLIN and OUT was the best combination to reduce VL vector densities for 22 months or longer. Operationally, this is much easier to apply than IRS. A cost analysis of the preferred tools will follow. The relationship between vector density (males plus females) and leishmaniasis incidence should be investigated, and this will require estimates of the Entomological Inoculation Rate.

Heart Failure Induced by Perinatal Ablation of Cardiac Myosin Light Chain Kinase.

Background: Germline knockout mice are invaluable in understanding the function of the targeted genes. Sometimes, however, unexpected phenotypes are encountered, due in part to the activation of compensatory mechanisms. Germline ablation of cardiac myosin light chain kinase (cMLCK) causes mild cardiac dysfunction with cardiomyocyte hypertrophy, whereas ablation in adult hearts results in acute heart failure with cardiomyocyte atrophy. We hypothesized that compensation after ablation of cMLCK is dependent on developmental staging and perinatal-onset of cMLCK ablation will result in more evident heart failure than germline ablation, but less profound when compared to adult-onset ablation. Methods and Results: The floxed-Mylk3 gene was ablated at the beginning of the perinatal stage using a single intra-peritoneal tamoxifen injection of 50 mg/kg into pregnant mice on the 19th day of gestation, this being the final day of gestation. The level of cMLCK protein level could no longer be detected 3 days after the injection, with these mice hereafter denoted as the perinatal Mylk3-KO. At postnatal day 19, shortly before weaning age, these mice showed reduced cardiac contractility with a fractional shortening 22.8 ± 1.0% (n = 7) as opposed to 31.4 ± 1.0% (n = 11) in controls. The ratio of the heart weight relative to body weight was significantly increased at 6.68 ± 0.28 mg/g (n = 12) relative to the two control groups, 5.90 ± 0.16 (flox/flox, n = 11) and 5.81 ± 0.33 (wild/wild/Cre, n = 5), accompanied by reduced body weight. Furthermore, their cardiomyocytes were elongated without thickening, with a long-axis of 101.8 ± 2.4 µm (n = 320) as opposed to 87.1 ± 1.6 µm (n = 360) in the controls. Conclusion: Perinatal ablation of cMLCK produces an increase of heart weight/body weight ratio, a reduction of contractility, and an increase in the expression of fetal genes. The perinatal Mylk3-KO cardiomyocytes were elongated in the absence of thickening, differing from the compensatory hypertrophy shown in the germline knockout, and the cardomyocyte thinning shown in adult-inducible knockout.

Efficacy, Safety and Cost of Insecticide Treated Wall Lining, Insecticide Treated Bed Nets and Indoor Wall Wash with Lime for Visceral Leishmaniasis Vector Control in the Indian Sub-continent: A Multi-country Cluster Randomized Controlled Trial.

BACKGROUND: We investigated the efficacy, safety and cost of lime wash of household walls plus treatment of sand fly breeding places with bleach (i.e. environmental management or EM), insecticide impregnated durable wall lining (DWL), and bed net impregnation with slow release insecticide (ITN) for sand fly control in the Indian sub-continent. METHODS: This multi-country cluster randomized controlled trial had 24 clusters in each three sites with eight clusters per high, medium or low sand fly density stratum. Every cluster included 45-50 households. Five households from each cluster were randomly selected for entomological measurements including sand fly density and mortality at one, three, nine and twelve months post intervention. Household interviews were conducted for socioeconomic information and intervention acceptability assessment. Cost for each intervention was calculated. There was a control group without intervention. FINDINGS: Sand fly mortality [mean and 95%CI] ranged from 84% (81%-87%) at one month to 74% (71%-78%) at 12 months for DWL, 75% (71%-79%) at one month to 49% (43%-55%) at twelve months for ITN, and 44% (34%-53%) at one month to 22% (14%-29%) at twelve months for EM. Adjusted intervention effect on sand fly density measured by incidence rate ratio ranged from 0.28 (0.23-0.34) at one month to 0.62 (0.51-0.75) at 12 months for DWL; 0.72 (0.62-0.85) at one month to 1.02 (0.86-1.22) at 12 months for ITN; and 0.89 (0.76-1.03) at one months to 1.49 (1.26-1.74) at 12 months for EM. Household acceptance of EM was 74% compared to 94% for both DWL and ITN. Operational cost per household in USD was about 5, 8, and 2 for EM, DWL and ITN, respectively. Minimal adverse reactions were reported for EM and ITN while 36% of households with DWL reported transient itching. INTERPRETATION: DWL is the most effective, durable and acceptable control method followed by ITN. The Visceral Leishmaniasis (VL) Elimination Program in the Indian sub-continent should consider DWL and ITN for sand fly control in addition to IRS.

Implication of vector characteristics of Phlebotomus argentipes in the kala-azar elimination programme in the Indian sub-continent.

BACKGROUND: Visceral leishmaniasis (VL), also known as kala-azar in the Indian sub-continent (ISC), is a major public health concern in Bangladesh, India, and Nepal, where it is caused by Leishmania donovani transmitted by the sand fly Phlebotomus argentipes. Various ecological parameters including air temperature, rainfall, wind speed, relative humidity, soil moisture, pH, and organic carbon are known to influence the oviposition of female sand flies, as well as the survival and development of larvae. However, more detailed knowledge on vector behavior, such as biting times, breeding places, and preferred hosts are needed to design optimal evidence-based vector control interventions. METHODS: In order to facilitate rational decisions regarding VL vector control, a systematic review was conducted to identify the prevailing practice and knowledge gaps in relation to vector bionomics and behavior. Search terms included 'sand fly bionomics', 'habitat', and 'visceral leishmaniasis/kala-azar vector control' using the Boolean operator AND to identify the country of interest, namely: Bangladesh, India, and Nepal. Both PubMed and Google search engines were used. Additional unpublished documents in the three countries were also analyzed. RESULTS: Information on the life cycle of VL vectors, their breeding behavior, infection rate with L. donovani, feeding behavior, and seasonal variation are useful for designing vector control operations. Unfortunately, none of the studies on the life cycle of P. argentipes was conducted in field settings of the ISC, so the publications from other locations had to be used for determining the duration of life cycle and development from egg to adult. However, information about breeding places, seasonal variation of vector densities, and 47 out of the selected 51 papers are available from the ISC and can be used for intelligent design of control operations. CONCLUSION: Vector control services should undertake routine insecticide resistance monitoring and adapt indoor residual spraying rounds to the seasonality of vector densities. Further research is needed on potential animal reservoirs for L. donovani, on the breeding habitat, and life cycle of sand flies in the ISC.

Acute heart failure with cardiomyocyte atrophy induced in adult mice by ablation of cardiac myosin light chain kinase.

AIMS: Under pressure overload, initial adaptive hypertrophy of the heart is followed by cardiomyocyte elongation, reduced contractile force, and failure. The mechanisms governing the transition to failure are not fully understood. Pressure overload reduced cardiac myosin light chain kinase (cMLCK) by ∼80% within 1 week and persists. Knockdown of cMLCK in cardiomyocytes resulted in reduced cardiac contractility and sarcomere disorganization. Thus, we hypothesized that acute reduction of cMLCK may be causative for reduced contractility and cardiomyocyte remodelling during the transition from compensated to decompensated cardiac hypertrophy. METHODS AND RESULTS: To mimic acute cMLCK reduction in adult hearts, the floxed-Mylk3 gene that encodes cMLCK was inducibly ablated in Mylk3(flox/flox)/merCremer mice (Mylk3-KO), and compared with two control mice (Mylk3(flox/flox) and Mylk3(+/+)/merCremer) following tamoxifen injection (50 mg/kg/day, 2 consecutive days). In Mylk3-KO mice, reduction of cMLCK protein was evident by 4 days, with a decline to below the level of detection by 6 days. By 7 days, these mice exhibited heart failure, with reduction of fractional shortening compared with those in two control groups (19.8 vs. 28.0% and 27.7%). Severely convoluted cardiomyocytes with sarcomeric disorganization, wavy fibres, and cell death were demonstrated in Mylk3-KO mice. The cardiomyocytes were also unable to thicken adaptively to pressure overload. CONCLUSION: Our results, using a new mouse model mimicking an acute reduction of cMLCK, suggest that cMLCK plays a pivotal role in the transition from compensated to decompensated hypertrophy via sarcomeric disorganization.

Quantification of the natural history of visceral leishmaniasis and consequences for control.

BACKGROUND: Visceral leishmaniasis has been targeted for elimination as a public health problem (less than 1 case per 10,000 people per year) in the Indian sub-continent by 2017. However, there is still a high degree of uncertainty about the natural history of the disease, in particular about the duration of asymptomatic infection and the proportion of asymptomatically infected individuals that develop clinical visceral leishmaniasis. Quantifying these aspects of the disease is key for guiding efforts to eliminate visceral leishmaniasis and maintaining elimination once it is reached. METHODS: Data from a detailed epidemiological study in Bangladesh in 2002-2004 was analysed to estimate key epidemiological parameters. The role of diagnostics in determining the probability and rate of progression to clinical disease was estimated by fitting Cox proportional hazards models. A multi-state Markov model of the natural history of visceral leishmaniasis was fitted to the data to estimate the asymptomatic infection period and the proportion of asymptomatic individuals going on to develop clinical symptoms. RESULTS: At the time of the study, individuals were taking several months to be diagnosed with visceral leishmaniasis, leading to many opportunities for ongoing transmission. The probability of progression to clinical disease was strongly associated with initial seropositivity and even more strongly with seroconversion, with most clinical symptoms developing within a year. The estimated average durations of asymptomatic infection and symptomatic infection for our model of the natural history are 147 days (95 % CI 130-166) and 140 days (95 % CI 123-160), respectively, and are significantly longer than previously reported estimates. We estimate from the data that 14.7 % (95 % CI 12.6-20.0 %) of asymptomatic individuals develop clinical symptoms-a greater proportion than previously estimated. CONCLUSIONS: Extended periods of asymptomatic infection could be important for visceral leishmaniasis transmission, but this depends critically on the relative infectivity of asymptomatic and symptomatic individuals to sandflies. These estimates could be informed by similar analysis of other datasets. Our results highlight the importance of reducing times from onset of symptoms to diagnosis and treatment to reduce opportunities for transmission.

Mouse Model of Human Congenital Heart Disease: Progressive Atrioventricular Block Induced by a Heterozygous Nkx2-5 Homeodomain Missense Mutation.

BACKGROUND: Heterozygous human NKX2-5 homeodomain (DNA-binding domain) missense mutations are highly penetrant for varied congenital heart defects, including progressive atrioventricular (AV) block requiring pacemaker implantation. We recently replicated this genetic defect in a murine knockin model, in which we demonstrated highly penetrant, pleiotropic cardiac anomalies. In this study, we examined postnatal AV conduction in the knockin mice. METHODS AND RESULTS: A murine knockin model (Arg52Gly, Nkx2-5(+/R52G)) in a 129/Sv background was analyzed by histopathology, surface, and telemetry ECG, and in vivo electrophysiology studies, comparing with control Nkx2-5(+/+) mice at diverse postnatal stages, ranging from postnatal day 1 (P1) to 17 months. PR prolongation (first degree AV block) was present at 4 weeks, 7 months, and 17 months of age, but not at P1 in the mutant mice. Advanced AV block was also occasionally demonstrated in the mutant mice. Electrophysiology studies showed that AV nodal function and right ventricular effective refractory period were impaired in the mutant mice, whereas sinus nodal function was not affected. AV nodal size was significantly smaller in the mutant mice than their controls at 4 weeks of age, corresponding to the presence of PR prolongation, but not P1, suggesting, at least in part, that the conduction abnormalities are the result of a morphologically atrophic AV node. CONCLUSIONS: The highly penetrant and progressive AV block phenotype seen in human heterozygous missense mutations in NKX2-5 homeodomain was replicated in mice by knocking in a comparable missense mutation.

How dengue vector Aedes albopictus (Diptera: Culicidae) survive during the dry season in Dhaka City, Bangladesh?

BACKGROUND & OBJECTIVES: In 2000, a dengue outbreak occurred in Bangladesh that included Dhaka City. Both dengue vectors, Aedes aegypti and Ae. albopictus are present in Bangladesh. Aedes aegypti mosquitoes mainly breed in and around houses and Ae. albopictus is an outside breeder. There are many old trees throughout Dhaka City in different parks, streets and the university campus which may have holes that can contribute as potential breeding habitat for the dengue vector. Therefore, a survey was conducted to investigate the presence of eggs of the dengue vector mosquitoes in treeholes during the dry season in February 2001 to know their contribution on dengue outbreaks. METHODS: All treeholes in 10 different localities (parks, streets and university campus) of Dhaka City were surveyed. All trees were examined for treeholes up to the height of approximately 3 m and sampled. Debris were collected and packed in poly bags and brought to the laboratory for detailed studies. These were then soaked with tap water to observe egg hatching. The soaked materials were kept up to 20 days covered by a fine mosquito net. After 2-3 days, the eggs started hatching and larvae were separated from the sample for rearing up to IV instar. RESULTS: A total of 245 treeholes were surveyed in 49 identified tree species and 18 unidentified trees. Altogether, 1365 Aedes larvae were found, of which 1096 were Aedes albopictus and 269 were other Aedes species. The largest number of larvae was observed in Delonix regia of Leguminosae family. The number of Aedes albopictus found in the treeholes have perfect positive correlation with the number of other Aedes species. Not a single egg of Aedes aegypti was found in this survey. INTERPRETATION & CONCLUSION: This information will inform public health workers as well as the national control programme to help to solve mosquito borne diseases specially that of dengue. This is critical in planning for vector control operations due to the diversity of dengue outbreak in the nature.

How far are we from visceral leishmaniasis elimination in Bangladesh? An assessment of epidemiological surveillance data.

INTRODUCTION: In 2005, Bangladesh, India, and Nepal joined forces to eliminate Visceral Leishmaniasis (or kala-azar) from the region by 2015. In Bangladesh the elimination target is set at less than one new case per 10,000 population per year at upazila (sub-district) level. As the deadline approaches, we review the status of the elimination initiative in this country. METHODS: We collected all available disease surveillance data at the Disease Control Unit of the Directorate General of Health Services, Government of Bangladesh from 1994 to 2013. Additionally, we retrieved data from the Civil Surgeon Office from the Mymensingh district, one of the most heavily affected areas in Bangladesh. RESULTS: Between 1994 and 2013, 109,266 kala-azar cases causing 329 deaths were reported from 37 endemic districts in Bangladesh. Only 16 districts reported cases every year. The Mymensingh district was the most affected with 53,582 (49.04%) cases. Between 2008 and 2013 only 16 upazilas showed incidence rates above the elimination target in which they ranged from 1.06 to 18.25 per 10,000 people per year. DISCUSSION: While clear progress has been made towards eliminating VL, 16 upazilas in Bangladesh had not yet reached the target in 2013, based on official notification data that probably suffered from under-reporting bias. The elimination initiative urgently needs to establish methods to ascertain and monitor the elimination target.

Visceral leishmaniasis-associated mortality in Bangladesh: a retrospective cross-sectional study.

OBJECTIVE: To investigate visceral leishmaniasis (VL) deaths and risk factors in two VL endemic areas of Bangladesh. DESIGN: Retrospective cross-sectional. SETTING: Two geographically and culturally different VL endemic subdistricts, Godagari in the district of Rajshahi and Trishal in the district of Mymensingh in Bangladesh, August 2009-December 2011. PARTICIPANTS: 51 094 inhabitants from randomly selected Unions in the two subdistricts. MAIN OUTCOME MEASURES: VL deaths, confirmed independently by qualified physicians using the verbal autopsy procedure ICD10 guideline. RESULTS: The total number of people screened for VL deaths was 51 094 from 12 032 households from Godagari and Trishal subdistricts . About 16% of the people from Godagari were Tribals. The average age of the study population was 25.6 years (SD 18.4) and 49.7% were females. The VL case fatality rate averaged 6.12% (12/196) including 2/137 in Trishal and 10/59 in Godagari. Most of the VL deaths (9/12, 75%) occurred at home and the rest in tertiary hospitals. None of these deaths had been reported in the national VL surveillance system. The VL case fatality rate in the Tribal ethnic (22.2%) population was about 17 times higher than that in the Bangali ethnic (1.3%) population (p<0.0001). Tribal ethnicity had an 18 times (OR=18.1, 95% CI 3.6 to 90.6) higher risk for VL deaths compared with Bangali ethnicity (p<0.0001). CONCLUSION: VL deaths were found to be high in the study areas and were under-reported. The Tribal ethnic population was at the highest risk for VL deaths. The national VL Elimination Programme should give special attention to the tribal community in the endemic areas, especially for those in Rajshahi, and should strengthen VL surveillance by including tertiary hospitals in the national surveillance system.