Dimensionality reduction methods for extracting functional networks from large-scale CRISPR screens.

CRISPR-Cas9 screens facilitate the discovery of gene functional relationships and phenotype-specific dependencies. The Cancer Dependency Map (DepMap) is the largest compendium of whole-genome CRISPR screens aimed at identifying cancer-specific genetic dependencies across human cell lines. A mitochondria-associated bias has been previously reported to mask signals for genes involved in other functions, and thus, methods for normalizing this dominant signal to improve co-essentiality networks are of interest. In this study, we explore three unsupervised dimensionality reduction methods-autoencoders, robust, and classical principal component analyses (PCA)-for normalizing the DepMap to improve functional networks extracted from these data. We propose a novel "onion" normalization technique to combine several normalized data layers into a single network. Benchmarking analyses reveal that robust PCA combined with onion normalization outperforms existing methods for normalizing the DepMap. Our work demonstrates the value of removing low-dimensional signals from the DepMap before constructing functional gene networks and provides generalizable dimensionality reduction-based normalization tools.

A method for benchmarking genetic screens reveals a predominant mitochondrial bias.

We present FLEX (Functional evaluation of experimental perturbations), a pipeline that leverages several functional annotation resources to establish reference standards for benchmarking human genome-wide CRISPR screen data and methods for analyzing them. FLEX provides a quantitative measurement of the functional information captured by a given gene-pair dataset and a means to explore the diversity of functions captured by the input dataset. We apply FLEX to analyze data from the diverse cell line screens generated by the DepMap project. We identify a predominant mitochondria-associated signal within co-essentiality networks derived from these data and explore the basis of this signal. Our analysis and time-resolved CRISPR screens in a single cell line suggest that the variable phenotypes associated with mitochondria genes across cells may reflect screen dynamics and protein stability effects rather than genetic dependencies. We characterize this functional bias and demonstrate its relevance for interpreting differential hits in any CRISPR screening context. More generally, we demonstrate the utility of the FLEX pipeline for performing robust comparative evaluations of CRISPR screens or methods for processing them.

Household food insecurity and unimproved toilet facilities associate with child morbidity: evidence from a cross-sectional study in Bangladesh.

BACKGROUND: Despite recent progress in rural economic development and food production, the prevalence of household food insecurity (FI) and use of unimproved toilet facilities are widespread in Bangladesh. Evidence regarding the consequencs of household FI and poor sanitation on child morbidity is scarce. This study aimed to understand the association of FI and unimproved toilet facility with morbidity status of under-5 children in Bangladesh. METHODS: We used data from a cross-sectional survey that was conducted as part of an evaluation of the Maternal, Infant and Young Child Nutrition (MIYCN) Program in 9 districts of Bangladesh. The study population included children aged 6-59 months and their caregivers, identified using a two-stage cluster-sampling procedure. Child morbidity status was the outcome variable, and household FI status and type of toilet used were considered the main exposure variables in this study. We performed logistic regression, calculated adjusted odds ratios (AOR) to assess the association of child morbidity with household FI and unimproved toilet facility after adjusting for potential confounders. RESULTS: A total of 1,728 households were eligible for this analysis. About 23% of the households were food-insecure, and a large number of households had improved toilet facilities (93.4%). In the multivariable logistic regression model, we found that children in food-insecure households with unimproved toilet facility had 5.88 (AOR: 5.88; 95% CI 2.52, 13.70) times more chance, of being morbid compared to the children of food-secure households with improved toilet facility. A similar association of FI and toilet facilities with each of the morbidity components was observed, including diarrhea (AOR:3.6; 95% CI 1.79, 7.89), fever (AOR:3.47; 95% CI 1.72, 6.99), difficult or fast breathing with cough (AOR:3.88; 95% CI 1.99, 7.59), and difficult or fast breathing with blocked or running nose (AOR:1.29; 95% CI 0.56, 2.95). CONCLUSIONS: Our study shows that household FI and unimproved toilet facility jointly have more deteriorative effects on child morbidity than either of these conditions alone. Therefore, it is recommended to consider these two critical factors while designing a public health intervention for reducing morbidity among under-five children.

Biological and biorational management of blast diseases in cereals caused by Magnaporthe oryzae.

Blast diseases, caused by the fungal pathogen Magnaporthe oryzae, are among the most destructive diseases that occur on at least 50 species of grasses, including cultivated cereals wheat, and rice. Although fungicidal control of blast diseases has widely been researched, development of resistance of the pathogen against commercially available products makes this approach unreliable. Novel approaches such as the application of biopesticides against the blast fungus are needed for sustainable management of this economically important disease. Antagonistic microorganisms, such as fungi and probiotic bacteria from diverse taxonomic genera were found to suppress blast fungi both in vitro and in vivo. Various classes of secondary metabolites, such as alkaloids, phenolics, and terpenoids of plant and microbial origin significantly inhibit fungal growth and may also be effective in managing blast diseases. Common modes of action of microbial biocontrol agents include: antibiosis, production of lytic enzymes, induction of systemic resistance in host plant, and competition for nutrients or space. However, the precise mechanism of biocontrol of the blast fungus by antagonistic microorganisms and/or their bioactive secondary metabolites is not well understood. Commercial formulations of biocontrol agents and bioactive natural products could be cost-effective and sustainable but their availability at this time is extremely limited. This review updates our knowledge on the infection pathway of the wheat blast fungus, catalogs naturally occurring biocontrol agents that may be effective against blast diseases, and discusses their role in sustainable management of the disease.

Use of concurrent evaluation to improve implementation of a home fortification programme in Bangladesh: a methodological innovation.

OBJECTIVE: This paper focuses on the use of 'concurrent evaluation' to evaluate a nationally scaled-up programme in Bangladesh that was implemented by BRAC (an international development organisation) using Shasthya Shebika (SS) - volunteer community health workers - to promote home fortification with micronutrient powders (MNP) for children under-five. DESIGN: We developed a programme impact pathway to conceptualise the implementation and evaluation strategy and developed a strategic partnership among the key programme stakeholders for better use of evaluation evidence. We developed a multi-method concurrent evaluation strategy to provide insights into the BRAC programme and created provision for course correction to the implementation plan while it was in operation. SETTING: One hundred sixty-four sub-districts and six urban slums in Bangladesh. PARTICIPANTS: Caregivers of children 6-59 months, SS and BRAC's staff members. RESULTS: The evaluation identified low awareness about home fortification among caregivers, inadequate supply and frequent MNP stockouts, and inadequate skills of BRAC's SS to promote MNP at the community level as hindrances to the achievement of programme goals. The partners regularly discussed evaluation results during and after implementation activities to assess progress in programme coverage and any needs for modification. BRAC initiated a series of corrections to the original implementation plan to address these challenges, which improved the design of the MNP programme; this resulted in enhanced programme outcomes. CONCLUSIONS: Concurrent evaluation is an innovative approach to evaluate complex real-world programmes. Here it was utilised in implementing a large-scale nutrition programme to measure implementation process and effectiveness.

Cost-effectiveness of a market-based home fortification of food with micronutrient powder programme in Bangladesh.

OBJECTIVE: We estimated the cost-effectiveness of home fortification with micronutrient powder delivered in a sales-based programme in reducing the prevalence of Fe deficiency anaemia among children 6-59 months in Bangladesh. DESIGN: Cross-sectional interviews with local and central-level programme staff and document reviews were conducted. Using an activity-based costing approach, we estimated start-up and implementation costs of the programme. The incremental cost per anaemia case averted and disability-adjusted life years (DALY) averted were estimated by comparing the home fortification programme and no intervention scenarios. SETTING: The home fortification programme was implemented in 164 upazilas (sub-districts) in Bangladesh. PARTICIPANTS: Caregivers of child 6-59 months and BRAC staff members including community health workers were the participants for this study. RESULTS: The home fortification programme had an estimated total start-up cost of 35·46 million BDT (456 thousand USD) and implementation cost of 1111·63 million BDT (14·12 million USD). The incremental cost per Fe deficiency anaemia case averted and per DALY averted was estimated to be 1749 BDT (22·2 USD) and 12 558 BDT (159·3 USD), respectively. Considering per capita gross domestic product (1516·5 USD) as the cost-effectiveness threshold, the home fortification programme was highly cost-effective. The programme coverage and costs for nutritional counselling of the beneficiary were influential parameters for cost per DALY averted in the one-way sensitivity analysis. CONCLUSIONS: The market-based home fortification programme was a highly cost-effective mechanism for delivering micronutrients to a large number of children in Bangladesh. The policymakers should consider funding and sustaining large-scale sales-based micronutrient home fortification efforts assuming the clear population-level need and potential to benefit persists.

Role of home visits by volunteer community health workers: to improve the coverage of micronutrient powders in rural Bangladesh.

OBJECTIVE: We assessed the role of home visits by Shasthya Shebika (SS) - female volunteer community health workers (CHWs) - in improving the distribution of micronutrient powder (MNP), and explored the independent effects of caregiver-provider interaction on coverage variables. DESIGN: We used data from three cross-sectional surveys undertaken at baseline (n 1927), midline (n 1924) and endline (n 1540) as part of an evaluation of a home fortification programme. We defined an exposure group as one that had at least one SS visit to the caregiver's household in the 12 months preceding the survey considering three outcome variables - message (ever heard), contact (ever used) and effective coverage (regular used) of MNP. We performed multiple logistic regressions to explore the determinants of coverage, employed an 'interaction term' and calculated an odds ratio (OR) to assess the modifying effect of SS's home visits on coverage. SETTINGS: Sixty-eight sub-districts from ten districts of Bangladesh. PARTICIPANTS: Children aged 6-59 months and their caregivers. RESULTS: A home visit from an SS positively impacts message coverage at both midline (ratio of OR 1·70; 95 % CI 1·25, 2·32; P < 0·01) and endline (ratio of OR 3·58; 95 % CI 2·22, 5·78; P < 0·001), and contact coverage both at midline (ratio of OR 1·48; 95 % CI 1·06, 2·07; P = 0·021) and endline (ratio of OR 1·74; 95 % CI 1·23, 2·47; P = 0·002). There was no significant effect of a SS's home visit on effective coverage. CONCLUSIONS: The households visited by BRAC's volunteer CHWs have better message and contact coverage among the children aged 6-59 months.

What is the impact of removing performance-based financial incentives on community health worker motivation? A qualitative study from an infant and young child feeding program in Bangladesh.

BACKGROUND: Community health worker (CHW) motivation is an important factor related to health service quality and CHW program sustainability in low- and middle-income countries. Financial and non-financial motivators may influence CHW behavior through two dimensions of motivation: desire to perform and effort expended. The aim of this study was to explore how the removal of performance-based financial incentives impacted CHW motivation after formal funding ceased for Alive and Thrive (A&T), an infant and young child feeding (IYCF) program in Bangladesh. METHODS: This qualitative study included seven focus groups (n = 43 respondents) with paid supervisors of volunteer CHWs tasked with delivering interpersonal IYCF counseling services. Data were transcribed, translated into English, and then analyzed using both a priori themes and a grounded theory approach. RESULTS: Results suggest the removal of financial incentives was perceived to have negatively impacted CHWs' desire to perform in three primary ways: 1) a decreased desire to work without financial compensation, 2) changes in pre- and post-intervention motivation, and 3) household income challenges due to dependence on incentives. Removal of financial incentives was perceived to have negatively impacted CHWs' level of effort expended in four primary ways: 1) a reduction in CHW visits, 2) a reduction in quality of care, 3) CHW attrition, and 4) substitution of other income-generating activities. CONCLUSIONS: This study provides new evidence regarding how removing performance-based financial incentives from a CHW program can negatively impact CHW motivation. The findings suggest that program decision makers should consider how to construct community health work programs such that CHWs may continue to receive performance-based compensation after the original funding ceases.

Application of stacking hybrid machine learning algorithms in delineating multi-type flooding in Bangladesh.

Floods are among the most devastating natural hazards in Bangladesh. The country experiences multi-type floods (i.e., fluvial, flash, pluvial, and surge floods) every year. However, areas prone to multi-type floods have not yet been assessed on a national scale. Here, we used locally weighted linear regression (LWLR), random subspace (RSS), reduced error pruning tree (REPTree), random forest (RF), and M5P model tree algorithms in a hybrid ensemble to assess multi-type flood probabilities at a national scale in Bangladesh. We used historical flood data (1988-2020), remote sensing images (e.g., MODIS, Landsat 5-8, and Sentinel-1), and topography, hydrogeology, and environmental datasets to train and validate the proposed algorithms. According to the results, the stacking ensemble machine learning LWLR-RF algorithm performed better than the other algorithms in predicting flood probabilities, with R2 = 0.967-0.999, MAE = 0.022-0.117, RMSE = 0.029-0.148, RAE = 4.48-23.38%, and RRSE = 5.8829.69% for the training and testing datasets. Furthermore, true skill statistics (TSS: 0.929-0.967), corrected classified instances (CCI: 96.45-98.35), area under the curve (AUC: 0.983-0.997), and Gini coefficients (0.966-0.994) were computed to validate the constructed (LWLR-RF) multi-type flood probability maps. The maps constructed via the LWLR-RF algorithm revealed that the proportions of different categories of flooding areas in Bangladesh are fluvial flooding 1.50%, 5.71%, 12.66%, and 13.77% of the total land area; flash floods of 4.16%, 8.90%, 11.11%, and 5.07%; pluvial flooding: 5.72%, 3.25%, 5.07%, and 0.90%; and surge flooding, 1.69%, 1.04%, 0.52%, and 8.64% of the total land area, respectively. These percentages represent low, medium, high, and very high probabilities of flooding. The findings can guide future flood risk management and sustainable land-use planning in the study area.

Unintended consequences of programmatic changes to infant and young child feeding practices in Bangladesh.

BRAC, an international development organization, implemented a home-fortification programme from 2014 to 2018 in Bangladesh. This study aimed to understand the unintended consequences of programmatic changes that occurred during the implementation of the programme on the prevalence of good infant and young child feeding (IYCF) practices and other associated factors. We used pooled data from eight cross-sectional surveys and data from a series of qualitative investigations carried out as part of a mixed-methods evaluation approach. A total of 6,479 caregivers of children aged 6 to 23 months participated in the surveys. The prevalence of good IYCF practices increased from baseline (42.1%) to midline (45.3%), but it decreased at the endline survey (31.9%). Qualitative investigations identified several reasons for low IYCF practices at the programme level, such as the withdrawal of community health worker (CHW) incentives for promoting IYCF, providing incentives for the home-fortification of micronutrient powder (MNP) and changing the focus from IYCF promotion to MNP promotion. A multivariable generalized estimating equation model for pooled data revealed that caregivers were 28% (adjusted risk ratio [ARR]: 0.72, 95% CI [0.67, 0.78]) less likely to maintain good IYCF practices during the period when CHWs were not incentivized to promote IYCF compared to the period when CHWs were incentivized to promote it. The prevalence of good IYCF practices decreased from both baseline and midline to the endline survey due to the unintended consequences of the programmatic changes. An integrated intervention strategy to promote the home-fortification of MNP and IYCF could be helpful to avoid unintended negative consequences of programmatic changes.

Hydrogen peroxide detoxifying enzymes show different activity patterns in host and non-host plant interactions with Magnaporthe oryzae Triticum pathotype.

Wheat blast caused by the hemibiotroph fungal pathogen Magnaporthe oryzae Triticum (MoT) pathotype is a destructive disease of wheat in South America, Bangladesh and Zambia. This study aimed to determine and compare the activities of antioxidant enzymes in susceptible (wheat, maize, barley and swamp rice grass) and resistant (rice) plants when interacting with MoT. The activities of reactive oxygen species-detoxifying enzymes; catalase (CAT), ascorbate peroxidase (APX), glutathione peroxidase (GPX), glutathione S-transferase (GST), peroxidase (POX) were increased in all plants in response to MoT inoculation with a few exceptions. Interestingly, an early and very high activity of CAT was observed within 24 h after inoculation in wheat, barley, maize and swamp rice grass with lower H2O2 concentration. In contrast, an early and high accumulation of H2O2 was observed in rice at 48 hai with little CAT activity only at a later stage of MoT inoculation. The activities of APX, GST and POD were also high at an early stage of infection in rice. However, these enzymes activities were very high at a later stage in wheat, barley, maize and swamp rice grass. The activity of GPX gradually decreased with the increase of time in rice. Taken together, our results suggest that late and early inductions of most of the antioxidant enzyme activities occurs in susceptible and resistant plants, respectively. This study demonstrates some insights into physiological responses of host and non-host plants when interacting with the devastating wheat blast fungus MoT, which could be useful for developing blast resistant wheat.

Effect of gamma radiation on microbial load, physico-chemical and sensory characteristics of common spices for storage.

The effect of gamma radiation on the decontamination of microbial population, physico-chemical, radiation sensitivity and sensory characteristics of common spices for storage were evaluated. Spices were irradiated with gamma doses of 0 (as control), 2, 4, 6, 8 and 10 kGy, packed in the glass vials and stored at room temperature (22 ± 2°C) in the laboratory. In this research, Bacillus, Salmonella and Listeria species were identified in un-irradiated spice samples. Results also indicated that gamma radiation reduced the total microbial population compared to control and optimum gamma radiation doses (6 kGy for red chili and turmeric; 4 kGy for cumin, coriander, garlic and black pepper; 2 kGy for ginger powder samples) were identified for decontamination of the organisms in the studied spices. It was concluded that no significant differences before and after gamma radiation were observed in physico-chemical, nutritional and sensory properties but significantly changed in microbial load in spices samples.

Revisiting the plant growth-promoting rhizobacteria: lessons from the past and objectives for the future.

Plant beneficial rhizobacteria (PBR) is a group of naturally occurring rhizospheric microbes that enhance nutrient availability and induce biotic and abiotic stress tolerance through a wide array of mechanisms to enhance agricultural sustainability. Application of PBR has the potential to reduce worldwide requirement of agricultural chemicals and improve agro-ecological sustainability. The PBR exert their beneficial effects in three major ways; (1) fix atmospheric nitrogen and synthesize specific compounds to promote plant growth, (2) solubilize essential mineral nutrients in soils for plant uptake, and (3) produce antimicrobial substances and induce systemic resistance in host plants to protect them from biotic and abiotic stresses. Application of PBR as suitable inoculants appears to be a viable alternative technology to synthetic fertilizers and pesticides. Furthermore, PBR enhance nutrient and water use efficiency, influence dynamics of mineral recycling, and tolerance of plants to other environmental stresses by improving health of soils. This report provides comprehensive reviews and discusses beneficial effects of PBR on plant and soil health. Considering their multitude of functions to improve plant and soil health, we propose to call the plant growth-promoting bacteria (PGPR) as PBR.

Can complex programs be sustained? A mixed methods sustainability evaluation of a national infant and young child feeding program in Bangladesh and Vietnam.

BACKGROUND: Poor early-life nutrition is a major barrier to good health and cognitive development, and is a global health priority. Alive & Thrive (A&T) was a multi-pronged initiative to improve infant and young child feeding behaviors. It aimed to achieve at-scale child health and nutrition improvements via a comprehensive approach that included nutrition counseling by health workers, policy change, social mobilization and mass media activities. This study evaluated the sustainability of activities introduced during A&T implementation (2009-2014) in Bangladesh and Vietnam. METHODS: This was a mixed methods study that used a quasi-experimental design. Quantitative data (surveys with 668 health workers, and 269 service observations) were collected in 2017; and analysis compared outcomes (primarily dose and fidelity of activities, and capacity) in former A&T intervention areas versus areas that did not receive the full A&T intervention. Additionally, we conducted interviews and focus groups with 218 stakeholders to explore their impressions about the determinants of sustainability, based on a multi-level conceptual framework. RESULTS: After program conclusion, stakeholders perceive declines in mass media campaigns, policy and advocacy activities, and social mobilization activities - but counseling activities were institutionalized and continued in both countries. Quantitative data show a persisting modest intervention effect: health workers in intervention areas had significantly higher child feeding knowledge, and in Bangladesh greater self-efficacy and job satisfaction, compared to their counterparts who did not receive the full package of A&T activities. While elements of the program were integrated into routine services, stakeholders noted dilution of the program focus due to competing priorities. Qualitative data suggest that some elements, such as training, monitoring, and evaluation, which were seen as essential to A&T's success, have declined in frequency, quality, coverage, or were eliminated altogether. CONCLUSIONS: The inclusion of multiple activities in A&T and efforts to integrate the program into existing institutions were seen as crucial to its success but also made it difficult to sustain, particularly given unstable financial support and human resource constraints. Future complex programs should carefully plan for institutionalization in advance of the program by cultivating champions across the health system, and designing unique and complementary roles for all stakeholders including donors.

Understanding key drivers and barriers to implementation of the WHO recommendations for the case management of childhood pneumonia and possible serious bacterial infection with amoxicillin dispersible tablets (DT) in Bangladesh: a qualitative study.

BACKGROUND: Pneumonia and possible serious bacterial infection (PSBI) are leading causes of death among under-five children. The World Health Organization (WHO) issued global recommendations for the case management of childhood pneumonia and PSBI when referral is not feasible with oral amoxicillin. However, few governments to date have incorporated child-friendly amoxicillin dispersible tablets (DT) into their national treatment guidelines and policies. We aimed to understand the key drivers to the implementation of WHO recommendations for childhood pneumonia and PSBI using amoxicillin DT in Bangladesh. METHODS: A qualitative study was conducted from October 2017 to March 2018 in two districts of Bangladesh. Interviews were completed with 67 participants consisting of government officials and key stakeholders, international development agencies, health service providers (HSPs), and caregivers of young children diagnosed and treated with amoxicillin for pneumonia or PSBI. Data were analyzed thematically. RESULTS: Policies and operational planning emerged as paramount to ensuring access to essential medicines for childhood pneumonia and PSBI. Though amoxicillin DT is included for National Newborn Health Programme and Integrated Management of Childhood Illnesses in the Operational Plan of the Directorate General of Health Services, inclusion in Community-Based Healthcare Project and Directorate General of Family Planning policies is imperative to securing national supply, access, and uptake. At the sub-national level, training on the use of amoxicillin DT as a first line intervention is lacking, resulting in inadequate management of childhood pneumonia by HSPs. Advocacy activities are needed to create community-wide demand among key stakeholders, HSPs, and caregivers not yet convinced that amoxicillin DT is the preferred formulation for the management of childhood pneumonia and PSBI. CONCLUSION: Challenges in policy and supply at the national level and HSP preparedness at the sub-national levels contribute to the slow adoption of WHO recommendations for amoxicillin DT in Bangladesh. A consultation meeting to disseminate study findings was instrumental in driving the development of recommendations by key stakeholders to address these challenges. A comprehensive and inclusive evidence-based strategy involving all divisions of the Ministry of Health and Family Welfare will be required to achieve national adoption of WHO recommendations and country-wide introduction of amoxicillin DT in Bangladesh.

First report of basal rot of dragon fruit caused by Fusarium oxysporum in Bangladesh.

Dragon fruit (Hylocereus polyrhizus) is a high value newly introduced fruit crop in Bangladesh. It has drawn considerable public attention due to its appealing flesh color, sweet taste and fruit qualities. Recently, basal rot of dragon fruit plants was observed in several farmer's fields, nurseries and in the research field of Bangabandhu Sheikh Mujibur Rahman Agricultural University (BSMRAU) where about 10-15% of plants were infected in each location. Initially, the symptoms appeared in the basal part near the soil as brown lesions which gradually extended to the upper stem and finally becoming soft and watery (Figure 1a). Infected plants were collected from Kapasia of Gazipur district (Latitude 24.266 and Longitude 90.633) to isolate the causal organism. Isolations were carried out following the procedure reported by Briste et al. (2019). Briefly, infected plant parts were surface sterilized in 2% NaOCl for 1 min followed by 70% ethanol for 5 min and rinsed 3 times with sterile double distilled water. A large piece of a surface sterilized plant was cut into small pieces (2 mm × 2 mm) from the margin of the necrotic lesion and placed on half strength potato dextrose agar (PDA) and incubated for 7 days at 25 °C. The BTFD1 and BTFD4 isolates were purified from single spores resulting in white colonies with a growth rate of 1cm/day on PDA (Figure 1b). Colonies produced single celled microconidia from unbranched, short monophialidic conidiophores and septate macroconidia as well as chlamydospores in PDA which is consistent with Fusarium oxysporum (Figure 1c). To confirm the identity of the isolates, the internal transcribed spacer (ITS1, 5.8S rRNA and ITS2) and translation elongation factor-1alpha (EF-1α) were amplified using primers ITS-1/ ITS-4 and EF1-728F/ EF1-986R, respectively (Surovy et al. 2018). The ITS sequences of the isolates BTFD1 and BTFD4 (GenBank accession # MN727096 and MN727095, respectively) showed 100% similarity with the sequence from F. oxysporum strain JJF2 (MN626452). Sequence identity for EF-1α (GenBank accession # MN752123 and MN752124, respectively) was 100% with the sequence from F. oxysporum strain CAV041_EO (MK783088). The isolates (BTFD1 and BTFD4) were identified as F. oxysporum based on the aligned sequences of ITS and EF-1α, molecular phylogenetic analyses by maximum likelihood tree (Figure 2a) and maximum parsimony tree methods (Figure 2b). The isolates were stored at 4°C on dried filter paper as well as in an ultra-low temperature freezer (-80°C) at IBGE, BSMRAU, Bangladesh and are available on request. To ensure pathogenicity, isolate BTFD1 was grown on PDA, incubated at 25°C for 7 days and 250 ml conidial suspension (with 1 × 105 conidia/ml) was prepared. Twelve,three-month-old healthy dragon fruit plants were inoculated. Pathogenicity tests were carried out in two sets using three replications in each set. In one set, only the basal part of the plants was dipped into the conidial suspension and in another set the whole plant was dipped into the conidial suspension for two hours. Sterile distilled water was also used in another set of plants as a control. The inoculated plants were placed on wet tissue in a plastic box (31cm × 24cm × 8cm) covered and incubated at 25°C. After 10 days, all inoculated plants in both sets developed rot symptoms similar to those observed in the field, while the control plants remained healthy (Figure 1d). The pathogen was successfully re-isolated from the inoculated symptomatic parts on half strength PDA medium and had morphology as characterized before, thus fulfilling Koch's postulates. This disease has been reported in Argentina and Malaysia (Wright et al. 2007; Hafifi et al. 2019). To the bet of our knowledge, this is the first report of Fusarium basal rot of dragon fruit in Bangladesh caused by F. oxysporum.

Does health worker performance affect clients' health behaviors? A multilevel analysis from Bangladesh.

BACKGROUND: Suboptimal healthcare quality may be a barrier to achieving child health improvements, yet little is known about the relationship between provider compliance with evidence-based practices and client behavior change. We assess provider compliance in the context of infant and young child feeding (IYCF) counseling, its relationship with client IYCF behaviors in Bangladesh, and explore its potential determinants. METHODS: We use data from a 2017 evaluation of an IYCF program that includes a health worker survey (n = 74), caregiver survey (n = 232), and direct service observation checklists of counseling sessions (n = 232 observations of 74 health workers). We assess the relationship between provider compliance with recommended IYCF counseling topics and behaviors (standardized to a 100-point scale) and three reported IYCF behaviors among clients using multi-level models with random effects at the health worker and sub-district (sampling) levels. We also evaluate whether health worker self-efficacy, satisfaction, and technical knowledge are associated with provider compliance. RESULTS: Health worker compliance was significantly associated with reported exclusive breastfeeding for children under 6 months of age (adjusted odds ratio per 1 percentage point increase in counseling compliance score = 1.06, 95% CI 1.01, 1.12) and marginally associated with minimum dietary diversity (adjusted odds ratio per 1 percentage point increase in counseling compliance score = 1.05, 95% CI 1.00, 1.11). Counseling compliance was significantly and positively associated with both health worker self-efficacy and technical knowledge. CONCLUSIONS: We find evidence for an association between health worker compliance and client health behaviors; however, small effect sizes suggest that behavior change is multifactorial and affected by factors beyond care quality. Improvements to technical quality of care may contribute to desired health outcomes; but policies and programs seeking to change health behaviors through counseling may also wish to target upstream factors such as self-efficacy, alongside technical skill-building and knowledge, for maximum impact.

Pseudomonas and Burkholderia inhibit growth and asexual development of Phytophthora capsici.

The objective of this study was to isolate and characterize antagonistic rhizobacteria from chili against a notorious phytopathogen Phytophthora capsici. Among the 48 bacteria isolated, BTLbbc-02, BTLbbc-03, and BTLbbc-05 were selected based on their inhibitory activity against P. capsici. They were tentatively identified as Burkholderia metallica BTLbbc-02, Burkholderia cepacia BTLbbc-03, and Pseudomonas aeruginosa BTLbbc-05, respectively, based on their 16S rRNA gene sequencing. All inhibited the growth of P. capsici at varying levels by inducing characteristic morphological alterations of P. capsici hyphae. The cell-free culture supernatant of all three isolates impaired motility (up to 100%) and caused lysis (up to 50%) of the halted zoospores. Bioassays revealed that Pseudomonas sp. had higher antagonism and zoospore motility-inhibitory effects against P. capsici compared with two other isolates, Burkholderia spp. and B. metallica, which caused vacuolation in mycelium. All three bacteria suppressed sporangium formation and zoosporogenesis of P. capsici, and improved the seed germination and growth of cucumber. Our findings suggest that epiphytic bacteria, B. metallica, B. cepacia, and P. aeruginosa, could be used as potential biocontrol agents against P. capsici. A further study is required to ensure conformity with the existing regulations for soil, plant, and human health.

Epidemiological Significance of Crown Rot in the Fruiting Field in Relation to Colletotrichum gloeosporioides Infection of Strawberry Nursery Plants.

Anthracnose crown rot (ACR), caused by Colletotrichum gloeosporioides, is a serious disease of strawberry (Fragaria × ananassa) in the southeastern United States, and there is a need to determine the link between nursery and fruiting-field disease risk. A 2-year study in 2007, repeated in 2008, was conducted at the North Carolina State University Horticultural Crops Research Station, Clinton, using 'Chandler', the most popular cultivar in North Carolina and one that is highly susceptible to ACR. Mother plants in a summer nursery were inoculated midseason with three pathogenic strains of C. gloeosporioides at an incidence level of 0, 5, 10, or 25%. Asymptomatic runner plants were selected at maturity (85 to 88 days after inoculation) from the nursery in early to mid-October from within a 0.5-m (inner) or 0.5- to 1.0-m (outer) radius around inoculated mother plants and planted into a plasticulture fruiting field system, with fruit harvest in April to June the following spring. Plants collected from the 25%-inner treatment had the greatest area under the disease progress curve (AUDPC) values, with a terminal ACR-related plant mortality of 32 and 20% by the end of fruit harvest and marketable yield losses of 30.5 and 30.2% in 2007-08 (Yr1) and 2008-09 (Yr2) seasons, respectively. All treatments increased AUDPC values compared with noninoculated treatments except the 10%-outer (O) and 5%-O treatments in Yr1 and Yr2, respectively. Marketable yield decreased 291.6 kg/ha for every percent increase in inoculum level (i.e., 0 to 25%, R2 = 0.696, P = 0.001). Levels of quiescent infection (QI) incidence (percentage of sampled leaves) assessed 25 to 28 days before digging runner plants also directly affected yield. For example, yield decreased 131.0 kg/ha for every percent increase in QI incidence in mother plants (R2 = 0.744, P = 0.001). Immersion of plants in fungicide solutions prior to planting decreased AUDPC values and improved plant stand by 7 to 11% but did not affect marketable yield compared with controls. This study provides results that can enable nursery and fruit growers to assess risk and implement mitigation measures to limit nursery plant and fruit yield losses.