Beyond the regulatory radar: knowledge and practices of rural medical practitioners in Bangladesh.

BACKGROUND: Informal and unregulated rural medical practitioners (RMPs) provide healthcare services to about two-thirds of people in Bangladesh, although their service is assumed to be substandard by qualified providers. As the RMPs are embedded in the local community and provide low-cost services, their practice pattern demands investigation to identify the shortfalls and design effective strategies to ameliorate the service. METHODS: We conducted a cross-sectional study in 2015-16 using a convenient sample from all 64 districts of Bangladesh. Personnel practising modern medicine, without any recognized training, or with recognized training but practising outside their defined roles, and without any regulatory oversight were invited to take part in the study. Appropriateness of the diagnosis and the rationality of antibiotic and other drug use were measured as per the Integrated Management of Childhood Illness guideline. RESULTS: We invited 1004 RMPs, of whom 877 consented. Among them, 656 (74.8%) RMPs owned a drugstore, 706 (78.2%) had formal education below higher secondary level, and 844 (96.2%) had informal training outside regulatory oversight during or after induction into the profession. The most common diseases encountered by them were common cold, pneumonia, and diarrhoea. 583 (66.5%) RMPs did not dispense any antibiotic for common cold symptoms. 59 (6.7%) and 64 (7.3%) of them could identify all main symptoms of pneumonia and diarrhoea, respectively. In pneumonia, 28 (3.2%) RMPs dispensed amoxicillin as first-line treatment, 819 (93.4%) dispensed different antibiotics including ceftriaxone, 721 (82.2%) dispensed salbutamol, and 278 (31.7%) dispensed steroid. In diarrhoea, 824 (94.0%) RMPs dispensed antibiotic, 937 (95.4%) dispensed ORS, 709 (80.8%) dispensed antiprotozoal, and 15 (1.7%) refrained from dispensing antibiotic and antiprotozoal together. CONCLUSIONS: Inappropriate diagnoses, irrational use of antibiotics and other drugs, and polypharmacy were observed in the practising pattern of RMPs. The government and other stakeholders should acknowledge them as crucial partners in the healthcare sector and consider ways to incorporate them into curative and preventive care.

Rapid and non-invasive multivariate approach for the quality control of raw milk from mountain areas based on proton transfer reaction mass spectrometry data.

RATIONALE: A rapid and non-invasive method for the control of milk by proton transfer reaction mass spectrometry was developed. The approach has the potential to verify the geographic origin and altitude of dairy farms, provided that the cows have been extensively grazed with forage that reflects the botanical composition of the mountain environment. METHODS: Over a 1-month period, a total of 116 samples were analysed by proton transfer reaction mass spectrometry (PTRMS). A multivariate control chart based on the Hotelling T2 statistic was built with PTRMS data and, for comparison, with the chemical parameters obtained by infrared spectroscopy (FTIR, MilkoScan). RESULTS: The headspace analysis of the samples led to characteristic volatile profiles. Farms located in different mountain areas were discriminated by the protonated molecules m/z 45 (acetaldehyde), 59 (acetone), 73 (2-butanone) and 89 (butyric acid, ethyl acetate, pentanol). Milk samples were also discriminated according to the altitude of the farms according to m/z 45, 59, 63 (dimethyl sulfide), 73 (propionic acid, methyl acetate) and 81 (terpenes). CONCLUSIONS: A multivariate control chart based on PTRMS data was used for the quality control of milk. Milk samples from farms located at different mountain areas and altitudes were successfully discriminated.

Effects of ovarian disaggregation on adult murine follicle yield and viability.

Follicles are isolated from ovaries for numerous reasons, including IVM, but adult murine yields are <2 folliclesmg-1. The aim of the present study was to optimise ovarian disaggregation and develop methods applicable to the rapid screening of follicle viability. Ovaries from adult mice (n=7) were halved and disaggregated mechanically, or by using collagenase IV (Col-IV; 590UmL-1) or animal origin-free collagenase IV (AOF) at 590 or 1180UmL-1. Isolated follicles were stained with 4',6'-diamidino-2-phenylindole (DAPI; nuclei), chloromethyl-X-rosamine (CMXRos; mitochondria) or fluorescein isothiocyanate-conjugated anti-α-tubulin antibody. Follicle diameters and staining were measured and analysed using ImageJ, and data analysed using GraphPad Prism. Col-IV disaggregation yielded the highest number of follicles (17±10 folliclesmg-1 ovarian tissue). All disaggregation methods released more secondary follicles (86±20 per ovary; P<0.05) than any other size cohort. Mechanical and Col-IV disaggregation yielded similar numbers of morphologically intact follicles, whereas AOF disaggregation caused more damage (P<0.01). As the morphological disruption increased, DAPI and CMXRos staining decreased (P<0.05), and tubulin localisation became more heterogeneous. Col-IV disaggregation gave the best yield of morphologically intact follicles containing viable granulosa cells. In conclusion, we improved adult murine follicle yields and applied molecular markers to assess follicle morphology, cellular cytoskeleton and mitochondrial function.

Hexanal as biomarker for milk oxidative stress induced by copper ions.

Milk flavor varies greatly due to oxidative stress during storage. Several studies have documented the use of volatile biomarkers for determining milk oxidation, but only a few have focused on the development of inline procedures enabling the monitoring of milk oxidative stress. In this work, oxidative stress was induced in pasteurized milk samples by spiking increasing concentrations of copper ions (from 0 to 32 mg·L-1). During storage (4°C), hexanal evolution was monitored by a proton transfer reaction mass spectrometer. The mass fragment m/z 83 was selected as a biomarker for hexanal determination. Its intensity evolved with a sigmoidal trend, showing a maximum rate proportional to the Cu2+ content in milk. The proposed approach is simple, fast (up to 120 sample/h), sensitive (8.8 µg·m-3 per µM hexanal in the sample), with low limit of detection (0.5 µM, determined as 3 times the standard deviation divided by the slope of a calibration line), precise (<6%), with good recovery (99-104%), and noninvasive. The method can be used for laboratory screening of milk susceptibility toward oxidation or for quality control in the processing line.

Perception and attitude of medical doctors in Dhaka, Bangladesh, with regard to Ayurvedic medicine.

The World Health Organization (WHO) Traditional Medicine Strategy (2014-2023) aimed to help member states promote the safe and effective use of traditional medicine. While economic conditions have markedly improved in Bangladesh, the country is experiencing significant public health problems. Because of limited medical resources, there is a strong incentive to enhance complementary and alternative medicine usage in Bangladesh. Therefore, this study aimed to confirm the perceptions and attitudes of medical doctors (MDs) in Dhaka, Bangladesh, with regard to Ayurvedic medicine (AM). A total number of 159 MDs in Dhaka were interviewed by face-to-face between February and June 2015. The study revealed that 62.0% of MDs had treated patients with AM and 55.3% believed that AM should be regarded as its own specialty, whereas 39.7% of MDs believed that AM should be part of the conventional medical curriculum and 32.7% thought that AM did not seem scientific. In terms of gender, 45.3% of male MDs agreed or strongly agreed that AM only had a placebo effect. On the other hand, 65.8% of female MDs disagreed or strongly disagreed it. In terms of age, 77.0% of MDs aged 36 or elder (elder MDs) believed they were more likely to recommend AM use and 80.3% of elder MDs believed that the government should encourage more initiatives to promote AM. To enhance AM use, scientifically robust information on the efficacy, safety and scientific basis of AM should be more effectively conveyed to male MDs.

Awareness and knowledge of HIV/AIDS among married women in rural Bangladesh and exposure to media: a secondary data analysis of the 2011 Bangladesh Demographic and Health Survey.

The aims of this study were to describe awareness and knowledge of HIV/AIDS among Bangladeshi married women in rural areas and to examine associations between exposure to mass media and their awareness and comprehensive knowledge of HIV/AIDS where mass media has been suggested to be vital sources of information. From the original dataset of the sixth Bangladesh Demographic Health Survey in 2011, the data of 11,570 rural married women aged 15-49 years old were extracted. Logistic regression analyses were performed to estimate odds ratios (ORs) and 95% confidence intervals (CIs). We found that approximately two-thirds of women (63.0%) aged 15-49 years had heard about HIV/AIDS. Exposure to each type of media was significantly associated with awareness of HIV/AIDS. Comparing to those who were not exposed to each of the investigated media, the adjusted ORs of comprehensive knowledge of HIV/AIDS were significantly high for those exposed to newspapers/magazines less than once a week (1.34, 95% CI 1.09-1.65), newspapers/ magazines at least once a week (1.44, 95% CI 1.07-1.94), television at least once a week (1.41, 95% CI 1.18-1.68). It was suggested that television can be utilized to increase awareness and comprehensive knowledge of HIV/AIDS through effective programs. Although the level of exposure was still low, significant associations between exposure to newspapers/magazines and comprehensive knowledge of HIV/AIDS suggested potential of written messages to promote knowledge of HIV/AIDS.

Impact of tannery effluents on the aquatic environment of the Buriganga River in Dhaka, Bangladesh.

This study presents an overview of the existence and effects of six heavy metals, chromium (Cr), lead (Pb), cadmium (Cd), mercury (Hg), manganese (Mn), and aluminum (Al), in tannery effluents released to the Buriganga River in Dhaka, Bangladesh. The pollutants were found in three different sources, such as effluents from tanneries, contaminated river water and three species of fish-climbing perch (Anabas testudineus), spotted snakehead (Channa punctata), and Black tilapia (Oreochromis mossambicus) caught from the river. Tannery effluents, water, and fish samples were collected from three different factories, five sample stations, and three different harvesting points, respectively. Effluents from all three factories contained significant amounts of heavy metals, especially Cr (374.19 ppm in average), whereas lesser amounts were found in the tissues of the three fish species studied. The trends in tissue elemental concentrations of fish were Cr > Pb > Al > Hg > Mn > Cd. In most cases (Cr, Cd, Mn, and Al), heavy metal concentrations were found to be greater in climbing perch than in Black tilapia and spotted snakehead. Although the river water contained high concentrations of harmful heavy metals, the fish species under study had concentrations well below the permissible Food and Agriculture Organization/World Health Organization levels for those metals and seemed to be safe for human consumption.

Identification of Differentially Expressed Genes and Protein-Protein Interaction in Patients With COVID-19 and Diabetes Peripheral Neuropathy: A Bioinformatics and System Biology Approach.

The coronavirus disease 2019 (COVID-19) pandemic has had a significant impact globally, resulting in a higher death toll and persistent health issues for survivors, particularly those with pre-existing medical conditions. Numerous studies have demonstrated a strong correlation between catastrophic COVID-19 results and diabetes. To gain deeper insights, we analysed the transcriptome dataset from COVID-19 and diabetic peripheral neuropathic patients. Using the R programming language, differentially expressed genes (DEGs) were identified and classified based on up and down regulations. The overlaps of DEGs were then explored between these groups. Functional annotation of those common DEGs was performed using Gene Ontology (GO), Kyoto Encyclopedia of Genes and Genomes (KEGG), Bio-Planet, Reactome, and Wiki pathways. A protein-protein interaction (PPI) network was created with bioinformatics tools to understand molecular interactions. Through topological analysis of the PPI network, we determined hub gene modules and explored gene regulatory networks (GRN). Furthermore, the study extended to suggesting potential drug molecules for the identified mutual DEG based on the comprehensive analysis. These approaches may contribute to understanding the molecular intricacies of COVID-19 in diabetic peripheral neuropathy patients through insights into potential therapeutic interventions.

Cryopreservation Techniques for Ram Sperm.

Germplasm storage and transportation in artificial insemination (AI) and other advanced technologies are facilitated by cryopreservation. In reproduction, the cryopreservation of sperm allows it to be transported across vast distances and used even after the sire's death. However, the technique of cryopreservation might damage sperm and limit their activity. Several cryobiological investigations have reported that the integrity of the sperm membrane is frequently involved in the physical and biological elements that affect sperm survival at low temperatures during the cryopreservation process. However, successful cryopreservation of ram sperm is still a work in progress because a considerable percentage of sperm do not survive the freezing and thawing process. Sperms are destroyed during cryopreservation of semen due to varying concentrations of cryoprotective chemicals and if semen is not cooled at optimal cooling rates. Hence, it is crucial to know the optimum cooling rates with freezing and thawing protocols for maximum recovery of viable and functional sperm cells for a successful cryo-freezing of ram spermatozoa. Therefore, the current study compiled and compared the research on the impact of different cryopreservation procedures, cooling rates, equilibration time, and thawing protocols on post-thaw ram semen quality.

Rab7 Investigation Insights into the Existence of White Spot Syndrome Virus in Crustaceans: An In Silico Approach.

In this study, previously published Rab7 sequences from National Center for Biotechnology Information (NCBI) have been investigated from chordates, mollusks, annelids, cnidarians, amphibians, priapulids, brachiopods, and arthropods including decapods and other groups. Among decapod crustacean isolates, amino acid variations were found in 13 locations. Penaeid shrimps had variations in positions 13 (I ⟶ J), 22 (T ⟶ A), 124 (G ⟶ X), and 149 (V ⟶ X) while interestingly the freshwater prawn and mitten crab both had amino acid substitutions in positions 87 (V ⟶ C) and 95 (T ⟶ S) along with the other disagreements in amino acid positions 178 (S ⟶ N), 201 (D ⟶ E), 181 (E ⟶ D), 182 (L ⟶ I), 183 (Y ⟶ G), 184 (N ⟶ H), and 198 (A ⟶ T). Among 100 isolates of Rab7 from organisms of various phyla, mutations were observed in several positions. These mutations caused variations in hydrophobicity and isoelectric point which impact the ligand-protein binding affinity. Some common mutations were found in the organisms of the same phylum and among different phyla. Homology modeling of Rab7 proteins from different organisms was done using SWISS-MODEL and validated further by developing Ramachandran plots. Protein-protein docking showed that active residues were there in the binding interfaces of Rab7 from organisms of seven different phyla and VP28 of WSSV. Similarities were observed in the Rab7-VP28 complexes in those selected organisms which differed from the Rab7-VP28 complex in the case of Penaeid shrimp. The findings of this study suggest that WSSV may exist in different marine organisms that have Rab7 protein and transmit to crustaceans like shrimps and crabs which are of commercial importance.

Partnerships in the introduction of new routine vaccines in Bangladesh: evidence from a prospective process evaluation.

OBJECTIVE: To assess the contribution of partners in the introduction of two new vaccines concurrently: pneumococcal 10-valent conjugate vaccine (PCV-10) and inactivated polio vaccine (IPV) into the routine Expanded Programme on Immunization (EPI) in Bangladesh. DESIGN: We conducted a prospective process evaluation that included the theory of change development, root cause analysis and in-depth investigation. As part of process tracking, we reviewed relevant documents, observed trainers' and vaccinators' training and key stakeholder meetings. We analysed the data thematically. SETTING: We purposively selected eight Upazila (subdistrict) and one city corporation covering nine districts and seven administrative divisions of Bangladesh. PARTICIPANTS: Nineteen national key informants were interviewed and 16 frontline health workers were invited to the group discussions considering their involvement in the vaccine introduction process. RESULTS: The EPI experienced several challenges during the joint introduction of PCV-10 and IPV, such as frequent changes in the vaccine introduction schedule, delays in budget allocation, vaccine supply shortage and higher wastage rates of IPV. EPI addressed these challenges in collaboration with its partners, that is, the World Health Organization (WHO) and United Nations Children's Fund (UNICEF), who provided technical assistance to develop a training curriculum and communication materials and enhanced demand generation at the community level. In addition, the WHO conducted a country readiness assessment for PCV-10, and UNICEF supported vaccine shipment. Other government ministries, City Corporations and municipalities also supported the EPI. CONCLUSIONS: The partnership among the EPI stakeholders effectively addressed various operational challenges during the joint introduction of PCV-10 and IPV helped strengthen Bangladesh's immunisation systems. These accomplishments are attributed to several factors that should be supported and strengthened for future vaccine introductions in Bangladesh and other low and-middle countries.

Hunting Bats for Human Consumption in Bangladesh.

Bats are important wildlife to their ecologic system, but they are also a zoonotic disease reservoir. Close bat-human interaction can lead to pathogen spillover. We conducted a qualitative study in two districts of Bangladesh and interviewed 30 bat hunters who hunt bats primarily for consumption, to understand the process and their reasons for hunting bats and their perceptions about bats and bat-borne disease. Most hunters catch bats during winter nights, using a net. Bat meat is used for household consumption, and the surplus is sold to cover household expenditures. They prepare the bat meat at home to sell it in their own and in neighboring communities. They also sell live bats to traditional healers. They report that the bat population has declined compared with 5 or 10 years ago, a decline they attribute to hunting and deforestation. Many have heard of a disease from bat-contaminated date palm sap but do not believe that bats can spread such disease to humans. Close bat-human interaction reported in this study pose a risk of pathogen spillover. Conservation initiatives have the potential to reduce such interaction and so both reduce disease risk and support the ecology.

Effect of Chemotherapeutics and Tocopherols on MCF-7 Breast Adenocarcinoma and KGN Ovarian Carcinoma Cell Lines In Vitro.

The combination of doxorubicin and cyclophosphamide commonly used to treat breast cancer can cause premature ovarian failure and infertility. α-Tocopherol is a potent antioxidant whereas γ-tocopherol causes apoptosis in a variety of cancer models in vitro including breast cancer. We hypothesised that the combination of doxorubicin (Dox) and 4-hydroperoxycyclophosphamide (4-Cyc) would be more cytotoxic in vitro than each agent alone, and that α-tocopherol would reduce and γ-tocopherol would augment the cytotoxicity of the combined chemotherapeutics. Human MCF-7 breast cancer and KGN ovarian cells were exposed to Dox, 4-Cyc, combined Dox and 4-Cyc, α-tocopherol, γ-tocopherol, or a combination of Dox and 4-Cyc with α-tocopherol or γ-tocopherol. Cell viability was assessed using a crystal violet assay according to four schedules: 24h exposure, 24h exposure + 24h culture in medium, 24h exposure + 48h culture in medium, or 72h continuous exposure. Supernatants from each separate KGN culture experiment (n=3) were examined using an estradiol ELISA. Dox was cytotoxic to both MCF-7 and KGN cells, but 4-Cyc only killed MCF-7 cells. γ-Tocopherol significantly decreased MCF-7 but not KGN cell viability. The combined chemotherapeutics and γ-tocopherol were more cytotoxic to MCF-7 than KGN cells, and α-tocopherol reduced the cytotoxicity of the combined chemotherapeutics towards KGN ovarian cells, but not MCF-7 cells. The addition of both γ-tocopherol and α-tocopherol to the chemotherapeutic combination of Dox and cyclophosphamide has the potential to increase in vitro chemotherapeutic efficacy against breast cancer cells whilst decreasing cytotoxicity towards ovarian granulosa cells.

Real time monitoring and quantification of reactive oxygen species in breast cancer cell line MCF-7 by 2',7'-dichlorofluorescin diacetate (DCFDA) assay.

The detection of reactive oxygen species (ROS) using 2',7'-dichlorofluorescin diacetate (DCFDA) is commonly performed by a single measurement of fluorescence but this fails to capture a profile of ROS generation over time. This study aimed to develop a real-time monitoring method to increase the utility of the assay, to incorporate cytotoxicity screening and to describe the combined effects of DCFDA and the ROS generator, Ter-butyl hydrogen peroxide (TBHP). Breast cancer MCF-7 cells were loaded with DCFDA (0-50 µM) for 45 min, and then exposed to TBHP (0-50 µM). Fluorescence was recorded according to three different schedules: every hour for 6 h, or once after 6 h or 24 h. Viability was assessed in a crystal violet assay and cell morphology was examined by microscopy. TBHP caused a time and dose-dependent increase in ROS and the magnitude of the fluorescent signal was affected by the loading concentration of DCFDA. Reading the fluorescence every hour for 6 h did not diminish the emission signal. The most sensitive and reliable combination for this ROS assay was 10 µM DCFDA with 25 µM TBHP; since higher concentrations of DCFDA compromised cell viability. In conclusion we adapted a single point ROS assay to enable production of a profile of ROS generation over an extended 6 h period, and related this to cell viability and morphology.

A microcalorimetry study on the oxidation of linoleic acid and the control of rancidity.

The oxidative deterioration of unsaturated lipids fatty acids is a great concern for the food industry as it is associated with the development of rancid off-odors and the reduction of food quality. This work describes the potential use of isothermal microcalorimetry to monitor the oxidation of linoleic acid, an important conjugated fatty acid responsible for the development of rancidity. The heat flow signal developed during the oxidation process reflects a multistep mechanism typical of radical chain reactions. From the analysis of the thermograms, it is possible to identify a period of the reaction that occurs with an order equal to 0.5. This period is correlated with the propagation period of the radical chain reaction. In addition, it allows detecting the early appearance of rancidity notes, as confirmed with the analysis of the samples headspace by Proton-Transfer-Reaction Mass Spectrometry (PTR-MS). The proposed approach was finally applied to compare the capacity of natural and synthetic antioxidants to inhibit the oxidation process of linoleic acid and the occurrence of rancidity. The results presented here show the advantages of microcalorimetry to study oxidation reactions and their control.

Factors Influencing the Prevalence of Stunting Among Children Aged Below Five Years in Bangladesh.

BACKGROUND: Poor nutrition during childhood impedes physical and mental development of children, which propagate the vicious cycle of intergenerational under nutrition. This paper is aimed at understanding the determinants of stunting among children aged 0 to 59 months in Bangladesh. METHODS: The study used Bangladesh Demographic and Health Survey 2011 data and a multistage stratified cluster-sampling design. Anthropometric data (for height and weight) were collected and analysis was limited to 7647 children. Multiple binary logistic regression analysis was performed to assess the association of stunting with potential socioeconomic and demographic factors. RESULTS: The prevalence of stunting has been found to be about 41% among children aged less than 60 months and higher in rural setting than in urban areas (43% vs 36%). Adjusted model revealed that several factors were influencing stunting. The children living in moderately food-insecure households had higher odds of becoming stunted (odds ratio [OR] = 1.27, 95% confidence interval [CI]: 1.05-1.54, P = .01) compared to the children living in food-secure households. The derived ORs of stunting for children delivered at institutions facilitated particularly by public (OR = 0.80, 95% CI: 0.67-0.96; P = .02) or private (OR = 0.81, 95% CI: 0.67-0.97; P = .02) sectors were less than for children delivered at home. Similarly, wealth index, exposure of mother to the mass media, age of child, size of child at birth, and parents' education were significantly associated with stunting. CONCLUSIONS: Moreover, the demographic characteristics and other indicators appeared to have significant influence in the prevalence of stunting. Public health programs are needed to avert the risk factors of stunting among children in Bangladesh.

A large-scale behavior change intervention to prevent Nipah transmission in Bangladesh: components and costs.

BACKGROUND: Nipah virus infection (NiV) is a bat-borne zoonosis transmitted to humans through consumption of NiV-contaminated raw date palm sap in Bangladesh. The objective of this analysis was to measure the cost of an NiV prevention intervention and estimate the cost of scaling it up to districts where spillover had been identified. METHODS: We implemented a behavior change communication intervention in two districts, testing different approaches to reduce the risk of NiV transmission using community mobilization, interpersonal communication, posters and TV public service announcements on local television during the 2012-2014 sap harvesting seasons. In one district, we implemented a "no raw sap" approach recommending to stop drinking raw date palm sap. In another district, we implemented an "only safe sap" approach, recommending to stop drinking raw date palm sap but offering the option of drinking safe sap. This is sap covered with a barrier, locally called bana, to interrupt bats' access during collection. We conducted surveys among randomly selected respondents two months after the intervention to measure the proportion of people reached. We used an activity-based costing method to calculate the cost of the intervention. RESULTS: The implementation cost of the "no raw sap" intervention was $30,000 and the "only safe sap" intervention was $55,000. The highest cost was conducting meetings and interpersonal communication efforts. The lowest cost was broadcasting the public service announcements on local TV channels. To scale up a similar intervention in 30 districts where NiV spillover has occurred, would cost between $2.6 and $3.5 million for one season. Placing the posters would cost $96,000 and only broadcasting the public service announcement through local channels in 30 districts would cost $26,000. CONCLUSIONS: Broadcasting a TV public service announcement is a potential low cost option to advance NiV prevention. It could be supplemented with posters and targeted interpersonal communication, in districts with a high risk of NiV spillover.

A Controlled Trial to Reduce the Risk of Human Nipah Virus Exposure in Bangladesh.

Human Nipah virus (NiV) infection, often fatal in Bangladesh, is primarily transmitted by drinking raw date palm sap contaminated by Pteropus bats. We assessed the impact of a behavior change communication intervention on reducing consumption of potentially NiV-contaminated raw sap. During the 2012-2014 sap harvesting seasons, we implemented interventions in two areas and compared results with a control area. In one area, we disseminated a "do not drink raw sap" message and, in the other area, encouraged only drinking sap if it had been protected from bat contamination by a barrier ("only safe sap"). Post-intervention, 40% more respondents in both intervention areas reported knowing about a disease contracted through raw sap consumption compared with control. Reported raw sap consumption decreased in all areas. The reductions in the intervention areas were not significantly greater compared to the control. Respondents directly exposed to the "only safe sap" message were more likely to report consuming raw sap from a protected source than those with no exposure (25 vs. 15%, OR 2.0, 95% CI 1.5-2.6, P < 0.001). While the intervention increased knowledge in both intervention areas, the "only safe sap" intervention reduced exposure to potentially NiV-contaminated sap and should be considered for future dissemination.