Tailoring the Structure-Property Relationship of Ring-Opened Metathesis Copolymers for CO2-Selective Membranes.

In this work, we explore the use of ring-opening metathesis polymerization (ROMP) facilitated by a second-generation Grubbs catalyst (G2) for the development of advanced polymer membranes aimed at CO2 separation. By employing a novel copolymer blend incorporating 4,4'-oxidianiline (ODA), 1,6-hexanediamine (HDA), 1-adamantylamine (AA), and 3,6,9-trioxaundecylamine (TA), along with a CO2-selective poly(ethylene glycol)/poly(propylene glycol) copolymer (Jeffamine2003) and polydimethylsiloxane (PDMS) units, we have synthesized membranes under ambient conditions with exceptional CO2 separation capabilities. The strategic inclusion of PDMS, up to a 20% composition within the PEG/PPG matrix, has resulted in copolymer membranes that not only surpass the 2008 upper limit for CO2/N2 separation but also meet the commercial targets for CO2/H2 separation. Comprehensive analysis reveals that these membranes adhere to the mixing rule and exhibit percolation behavior across the entire range of compositions (0-100%), maintaining robust antiplasticization performance even under pressures up to 20 atm. Our findings underscore the potential of ROMP in creating precisely engineered membranes for efficient CO2 separation, paving the way for their application in large-scale environmental and industrial processes.

Developing Mixed Matrix Membranes with Good CO2 Separation Performance Based on PEG-Modified UiO-66 MOF and 6FDA-Durene Polyimide.

The use of mixed matrix membranes (MMMs) comprising metal-organic frameworks (MOFs) for the separation of CO2 from flue gas has gained recognition as an effective strategy for enhancing gas separation efficiency. When incorporating porous materials like MOFs into a polymeric matrix to create MMMs, the combined characteristics of each constituent typically manifest. Nevertheless, the inadequate dispersion of an inorganic MOF filler within an organic polymer matrix can compromise the compatibility between the filler and matrix. In this context, the aspiration is to develop an MMM that not only exhibits optimal interfacial compatibility between the polymer and filler but also delivers superior gas separation performance, specifically in the efficient extraction of CO2 from flue gas. In this study, we introduce a modification technique involving the grafting of poly(ethylene glycol) diglycidyl ether (PEGDE) onto a UiO-66-NH2 MOF filler (referred to as PEG-MOF), aimed at enhancing its compatibility with the 6FDA-durene matrix. Moreover, the inherent CO2-philic nature of PEGDE is anticipated to enhance the selectivity of CO2 over N2 and CH4. The resultant MMM, incorporating 10 wt% of PEG-MOF loading, exhibits a CO2 permeability of 1671.00 Barrer and a CO2/CH4 selectivity of 22.40. Notably, these values surpass the upper bound reported by Robeson in 2008.

Extended-Spectrum ß-Lactamases (ESBL): Challenges and Opportunities.

The rise of antimicrobial resistance, particularly from extended-spectrum ß-lactamase producing Enterobacteriaceae (ESBL-E), poses a significant global health challenge as it frequently causes the failure of empirical antibiotic therapy, leading to morbidity and mortality. The E. coli- and K. pneumoniae-derived CTX-M genotype is one of the major types of ESBL. Mobile genetic elements (MGEs) are involved in spreading ESBL genes among the bacterial population. Due to the rapidly evolving nature of ESBL-E, there is a lack of specific standard examination methods. Carbapenem has been considered the drug of first choice against ESBL-E. However, carbapenem-sparing strategies and alternative treatment options are needed due to the emergence of carbapenem resistance. In South Asian countries, the irrational use of antibiotics might have played a significant role in aggravating the problem of ESBL-induced AMR. Superbugs showing resistance to last-resort antibiotics carbapenem and colistin have been reported in South Asian regions, indicating a future bleak picture if no urgent action is taken. To counteract the crisis, we need rapid diagnostic tools along with efficient treatment options. Detailed studies on ESBL and the implementation of the One Health approach including systematic surveillance across the public and animal health sectors are strongly recommended. This review provides an overview of the background, associated risk factors, transmission, and therapy of ESBL with a focus on the current situation and future threat in the developing countries of the South Asian region and beyond.

Colletotrichum truncatum Causing Anthracnose of Tomato (Solanum lycopersicum L.) in Malaysia.

Tomato (Solanum lycopersicum L.) is a popular nutritious vegetable crop grown in Malaysia and other parts of the world. However, fungal diseases such as anthracnose pose significant threats to tomato production by reducing the fruit quality and food value of tomato, resulting in lower market prices of the crop globally. In the present study, the etiology of tomato anthracnose was investigated in commercial tomato farms in Sabah, Malaysia. A total of 22 fungal isolates were obtained from anthracnosed tomato fruits and identified as Colletotrichum species, using morphological characteristics. The phylogenetic relationships of multiple gene sequence alignments such as internal transcribed spacer (ITS), ß-tubulin (tub2), glyceraldehyde 3-phosphate dehydrogenase (gapdh), actin (act), and calmodulin (cal), were adopted to accurately identify the Colletotrichum species as C. truncatum. The results of pathogenicity tests revealed that all C. truncatum isolates caused anthracnose disease symptoms on inoculated tomato fruits. To our knowledge, the present study is the first report of tomato anthracnose caused by C. truncatum in Malaysia. The findings of this study will be helpful in disease monitoring, and the development of strategies for effective control of anthracnose on tomato fruits.

Characterization of the Phase-Variable Autotransporter Lav Reveals a Role in Host Cell Adherence and Biofilm Formation in Nontypeable Haemophilus influenzae.

Lav is an autotransporter protein found in pathogenic Haemophilus and Neisseria species. Lav in nontypeable Haemophilus influenzae (NTHi) is phase-variable: the gene reversibly switches ON-OFF via changes in length of a locus-located GCAA(n) simple DNA sequence repeat tract. The expression status of lav was examined in carriage and invasive collections of NTHi, where it was predominantly not expressed (OFF). Phenotypic study showed lav expression (ON) results in increased adherence to human lung cells and denser biofilm formation. A survey of Haemophilus species genome sequences showed lav is present in ∼60% of NTHi strains, but lav is not present in most typeable H. influenzae strains. Sequence analysis revealed a total of five distinct variants of the Lav passenger domain present in Haemophilus spp., with these five variants showing a distinct lineage distribution. Determining the role of Lav in NTHi will help understand the role of this protein during distinct pathologies.

Semi-Scavenging Poultry as Carriers of Avian Influenza Genes.

Ducks are the natural reservoir of influenza A virus and the central host for the avian influenza virus (AIV) subtype H5N1, which is highly pathogenic. Semi-scavenging domestic ducks allow for the reemergence of new influenza subtypes which could be transmitted to humans. We collected 844 cloacal swabs from semi-scavenging ducks inhabiting seven migratory bird sanctuaries of Bangladesh for the molecular detection of avian influenza genes. We detected the matrix gene (M gene) using real-time RT-PCR (RT-qPCR). Subtyping of the AIV-positive samples was performed by RT-qPCR specific for H5, H7, and H9 genes. Out of 844 samples, 21 (2.488%) were positive for AIV. Subtyping of AIV positive samples (n = 21) revealed that nine samples (42.85%) were positive for the H9 subtype, five (23.80%) were positive for H5, and seven (33.33%) were negative for the three genes (H5, H7, and H9). We detected the same genes after propagating the virus in embryonated chicken eggs from positive samples. Semi-scavenging ducks could act as carriers of pathogenic AIV, including the less pathogenic H9 subtype. This can enhance the pathogenicity of the virus in ducks by reassortment. The large dataset presented in our study from seven areas should trigger further studies on AIV prevalence and ecology.

Mixed Matrix Membranes for Efficient CO2 Separation Using an Engineered UiO-66 MOF in a Pebax Polymer.

Mixed matrix membranes (MMMs) have attracted significant attention for overcoming the limitations of traditional polymeric membranes for gas separation through the improvement of both permeability and selectivity. However, the development of defect-free MMMs remains challenging due to the poor compatibility of the metal-organic framework (MOF) with the polymer matrix. Thus, we report a surface-modification strategy for a MOF through grafting of a polymer with intrinsic microporosity onto the surface of UiO-66-NH2. This method allows us to engineer the MOF-polymer interface in the MMMs using Pebax as a support. The insertion of a PIM structure onto the surface of UiO-66-NH2 provides additional molecular transport channels and enhances the CO2 transport by increasing the compatibility between the polymer and fillers for efficient gas separation. As a result, MMM with 1 wt% loading of PIM-grafted-MOF (PIM-g-MOF) exhibited very promising separation performance, with CO2 permeability of 247 Barrer and CO2/N2 selectivity of 56.1, which lies on the 2008 Robeson upper bound. Moreover, this MMM has excellent anti-aging properties for up to 240 days and improved mechanical properties (yield stress of 16.08 MPa, Young's modulus of 1.61 GPa, and 596.5% elongation at break).

PIM-PI-1 and Poly(ethylene glycol)/Poly(propylene glycol)-Based Mechanically Robust Copolyimide Membranes with High CO2-Selectivity and an Anti-aging Property: A Joint Experimental-Computational Exploration.

Polymer membranes with excellent thermomechanical properties and good gas separation performance are desirable for efficient CO2 separation. A series of copolyimide membranes are prepared for the first time using PIM-PI-1, a hard segment with high CO2 permeability, and poly(ethylene glycol)/poly(propylene glycol) (PEG/PPG), a soft segment with high CO2 selectivity. Two different unit polymers are combined to compensate the limitations of each polymer (e.g., the fast aging and moderate selectivity of PIM-PI-1 and the poor mechanical properties and lower permeability of PEG/PPG). The corresponding PIM-(durene-PEG/PPG) membranes exhibit an excellent combination of mechanical properties and gas separation performance compared to the typical PI-PEG-based copolymer membrane. The improved mechanical property is attributed to the unique chain threading and the reinforcement between the spiro unit of PIM and the flexible PEG/PPG at the molecular level, which has not previously been exploited for membranes. The PIM-(durene-PEG/PPG) membranes show a high CO2 permeability of 350-669 Barrer and a high CO2/N2 selectivity of 33.5-40.3. The experimental results are further evaluated with theoretical results obtained from molecular simulation studies, and a very good agreement between the experimental results and simulation results is found. Moreover, the PIM-(durene-PEG/PPG) copolymer membranes display excellent anti-aging performance for up to 1 year.

Prevalence and severity of gastro-intestinal parasites in buffalo calves at Sylhet division of Bangladesh.

Gastrointestinal (GI) parasites are one of the most widely recognized health problems of buffalo calves in Bangladesh. The study was undertaken to investigate the prevalence of GI parasitic infestation in buffalo calves at Sylhet division of Bangladesh. Fecal samples of 200 buffalo calves under 1 year of age (76 males and 124 females) were collected from Sylhet, Maulvibazar, Habiganj, and Sunamganj districts, and from each of the four regions, 50 samples were collected. Samples were examined through simple flotation, sedimentation, and McMaster techniques. The overall prevalence of GI parasites in Sylhet division was 65.5%. Among the samples, the highest prevalence of GI parasite was found in Sylhet (78%), followed by Sunamganj (68%), Maulvibazar (66%), and Habiganj (50%) districts. Among the identified GI parasites, the highest prevalence was observed in case of Neoascaris vitulorum (26.5%). The highest prevalence of GI parasite was observed in summer (69.84%), followed by rainy (69.62%) and winter (55.17%) seasons. Males and females were almost equally susceptible to GI parasitic infestation. Older calves (9-12 months, 77.14%) were most vulnerable to GI parasites. Egg per gram (EPG) of feces was additionally counted in the current study to know the severity of infestation. The range of EPG varied from 50 to 900 among the identified parasites. Among them, the highest count was found in the case of Neoascaris vitulorum (100-900). It may be concluded that aggregated policies and actions need to be taken to control the GI parasitic infestation in buffalo calves at Sylhet Division and elsewhere in developing countries like Bangladesh.

Development of CO2-Selective Polyimide-Based Gas Separation Membranes Using Crown Ether and Polydimethylsiloxane.

A series of CO2-selective polyimides (CE-PDMS-PI-x) was synthesized by copolymerizing crown ether diamine (trans-diamino-DB18C6) and PDMS-diamine with 4,4'-(hexafluoroisopropylidene) di-phthalic anhydride (6FDA) through the polycondensation reaction. The structural characteristics of the copolymers and corresponding membranes were characterized by nuclear magnetic resonance (NMR), infrared spectroscopy (ATR-FTIR), thermogravimetric analysis (TGA), differential scanning calorimetry (DSC), X-ray diffraction (XRD), and gel permeation chromatography (GPC). The effect of PDMS loading on the CE-PDMS-PI-x copolymers was further analyzed and a very good structure-property relationship was found. A well-distributed soft PDMS unit played a key role in the membrane's morphology, in which improved CO2-separation performance was observed at a low PDMS content (5 wt %). In contrast, the fine-grained phase separation adversely affected the separation behavior at a certain level of PDMS loading, and the PDMS was found to provide a flexible gas-diffusion path, affecting only the permeability without changing the selective gas-separation performance for the copolymers with a PDMS content of 20% or above.

Streptococcus suis Encodes Multiple Allelic Variants of a Phase-Variable Type III DNA Methyltransferase, ModS, That Control Distinct Phasevarions.

Streptococcus suis is a significant cause of bacterial meningitis in humans, particularly in Southeast Asia, and is a leading cause of respiratory and invasive disease in pigs. Phase-variable DNA methyltransferases, associated with restriction-modification (R-M) systems, are a source of epigenetic gene regulation, controlling the expression of multiple genes. These systems are known as phasevarions (phase-variable regulons) and have been characterized in many host-adapted bacterial pathogens. We recently described the presence of a Type III DNA methyltransferase in S. suis, ModS, which contains a simple sequence repeat (SSR) tract within the open reading frame of the modS gene and which differed in length between individual strains. We also observed that multiple allelic variants of the modS gene were present in a population of S. suis isolates. Here, we demonstrate that a biphasic ON-OFF switching of expression occurs in the two most common ModS alleles, ModS1 and ModS2, and that switching is dependent on SSR tract length. Furthermore, we show using single-molecule real-time (SMRT) sequencing that ModS1 and ModS2 are active methyltransferases in S. suis ON-OFF switching of each ModS allele results in the regulation of distinct phasevarions, with the ModS2 phasevarion impacting growth patterns and antibiotic resistance. This is the first demonstration of a phase-variable Type III DNA methyltransferase in a Gram-positive organism that controls a phasevarion. Characterizing the phenotypic effects of phasevarions in S. suis is key to understanding pathogenesis and the development of future vaccines.IMPORTANCEStreptococcus suis is a causative agent of meningitis, polyarthritis, and polyserositis in swine, and it is a major cause of zoonotic meningitis in humans. Here, we investigate epigenetic gene regulation in S. suis by multiple phasevarions controlled by the phase-variable Type III DNA methyltransferase ModS. This is the first characterized example of a Type III R-M system regulating a phasevarion in a Gram-positive organism. We demonstrate that biphasic ON-OFF switching of ModS expression results in differences in bacterial growth and antibiotic resistance. Understanding the effects of ModS phase variation is required to determine the stably expressed antigenic repertoire of S. suis, which will direct and inform the development of antimicrobial treatments and vaccines against this important pathogen.

Cross-Linked Mixed-Matrix Membranes Using Functionalized UiO-66-NH2 into PEG/PPG-PDMS-Based Rubbery Polymer for Efficient CO2 Separation.

Mixed-matrix membranes (MMMs) with an ideal polymer-filler interface and high gas separation performance are very challenging to fabricate because of incompatibility between the fillers and the polymer matrix. This work provides a simple technique to prepare a series of cross-linked MMMs (xMMM@n) by covalently attaching UiO-66-NB metal-organic frameworks (MOFs) within the PEG/PPG-PDMS copolymer matrix via ring-opening metathesis polymerization and in situ membrane casting. The norbornene-modified MOF (UiO-66-NB) is successfully copolymerized and dispersed homogeneously into a PEG/PPG-PDMS matrix because of very fast polymer formation and strong covalent interaction between MOFs and the rubbery polymer. A significant improvement in gas permeability is achieved in membranes up to a 5 wt % MOF loading compared to the pristine polymer membrane without affecting selectivity. The CO2/N2 separation performance of xMMM@1, xMMM@3, and xMMM@5 with 1, 3, and 5 wt % MOF loading, respectively, surpassed Robeson's 2008 upper bound. In addition, the best performing membrane, xMMM@3 (PCO2 = 585 Barrer and CO2/N2 ∼53), approaches the 2019 upper bound, indicating that the cross-linked MMMs (xMMM@n) are very promising for CO2 separation from flue gas. The experimental results of our study were evaluated and are supported by theoretical data obtained using the Maxwell model for MMMs. Moreover, the developed MMMs, xMMM@ns, displayed outstanding antiplasticization performance at pressures of up to 25 atm and very stable antiaging performance for up to 11 months with good temperature switching behaviors.

Isolation and molecular characterization of multidrug-resistant Escherichia coli from chicken meat.

Antibiotic-resistant Escherichia coli (E. coli) are common in retail poultry products. In this study, we aimed to isolate and characterize multidrug resistant (MDR) E. coli in raw chicken meat samples collected from poultry shops in Sylhet division, Bangladesh, as well as to determine correlation between resistance phenotype and genotype. A total of 600 chicken meat swabs (divided equally between broiler and layer farms, n = 300 each) were collected and the isolates identified as E. coli (n = 381) were selected. Disc diffusion antimicrobial susceptibility assay showed resistance of these isolates to ampicillin, erythromycin, tetracycline, streptomycin, trimethoprim-sulfamethoxazole, chloramphenicol, and gentamicin. Polymerase chain reaction (PCR) identified several antibiotic resistance genes (ARGs) in our isolates. Among these ARGs, the prevalence of tetA (for tetracycline) was the highest (72.58%) in broiler chicken isolates, followed by sul1 (for sulfonamide; 44.16%), aadA1 (for streptomycin; 33.50%), ereA (for erythromycin; 27.41%), aac-3-IV (for gentamicin; 25.38%), and the two genes cmlA (24.87%) and catA1 (8.63%) for chloramphenicol. On the other hand, the respective prevalence in layer chicken isolates were 82.06%, 47.83%, 35.87%, 35.33%, 23.91%, 19.02%, and 5.43%. Furthermore, 49.23% of the isolates from broiler chicken were MDR, with the presence of multiple antibiotic resistance genes, including 3 (40.11%) and 4 (9.13%) genes. On the other hand, 51.09% of layer chicken E. coli isolates were MDR, with 3, 4 or 5 ARGs detected in 36.41%, 14.13%, and 0.54% of the isolates, respectively. We also found that 12.8% of broiler chicken E. coli isolates and 7.61% of layer chicken isolates carried genes coding for extended-spectrum SHV beta-lactamases. Lastly, we report the presence of the AmpC beta-lactamase producing gene (CITM) in 4.56% and 3.26% of broiler and layer chicken E. coli isolates, respectively. We found significant correlations between most of the antimicrobial resistant phenotypes and genotypes observed among the investigated E. coli isolates. Our findings highlight the need for the prudent use of antimicrobials in chickens to minimize the development of antibiotic-resistant bacterial strains.

Integration of health education intervention to improve the compliance to mass drug administration for soil-transmitted helminths infection in Bangladesh: An implementation research.

INTRODUCTION: In Bangladesh, the prevention and control strategy of soil-transmitted helminthiasis (STHs) is based on the mass drug administration (MDA) program. Despite bi-annual MDA since 2008, the reported compliance is still below the target, and the STH prevalence is high in several areas. This study was done to assess the feasibility and barriers of integrating health education (HE) intervention to achieve the target MDA compliance in the local context of Bangladesh. MATERIALS AND METHODS: A mixed-method study, utilizing PRISM (Practical Robust Implementation Sustainability Model) framework, was conducted between July 2017 to March 2018 in Dhaka and Sylhet divisions of Bangladesh. A total of 640 school-aged children selected from four different schools were divided into intervention and control groups. Eight focus group discussions (FGDs) and eight in-depth interviews (IDIs) were also conducted among 56 adults, including parents of school-aged children, school teachers, and health officers. RESULTS: Quantitative findings revealed that HE intervention had a significant role (P < .05) to improve the mean knowledge score in the intervention group (3.35) compared to the control group (0.29). STH preventive behaviours and MDA participating attitudes were also significantly increased in the intervention group (P < .05) compared to the control group. Some of the major barriers associated with HE integration identified in the qualitative study were budget deficiencies, inadequate training of program implementers, and information gaps. In contrast, the school environment and positive community attitudes were observed as supportive factors for the integration of HE. CONCLUSION: Increased knowledge score and behaviour changes due to HE intervention demonstrated in this study hint that integration of HE with MDA is feasible and can be promising to promote MDA compliance and to reduce STH prevalence in this setting. However, the allocation of adequate budget, as well as coordination and collaboration with local political context, should be addressed for the sustainability of integration.

Susceptibility and Multidrug Resistance Patterns of Escherichia coli Isolated from Cloacal Swabs of Live Broiler Chickens in Bangladesh.

Antimicrobial resistance is a major health problem, particularly in developing countries like Bangladesh, where there is a paucity of information on resistance patterns and prevalence of antimicrobial determinants. Therefore, the aims of this study were to investigate the prevalence of resistance, including multi-drug resistance (MDR), and the associated genetic determinants in Escherichia coli isolates from cloacal swabs of live broiler chickens in Bangladesh. Altogether, 400 cloacal swabs (200 from Rajshahi and 200 from Dhaka divisions) were randomly collected from individual chickens in 50 broiler farms. E. coli was isolated and identified using conventional bacteriological culture and biochemical methods. The isolates were further confirmed using genus-specific 16S rRNAtargeted polymerase chain reaction (PCR) primers. Antimicrobial susceptibilities and MDR of the isolates against nine different antimicrobial agents (ampicillin, erythromycin, tetracycline, gentamicin, ciprofloxacin, levofloxacin, trimethoprim-sulfamethoxazole, colistin sulphate, and streptomycin) were determined using the Kirby-Bauer disc diffusion method. Resistance determinants of E. coli to ampicillin (blaTEM), streptomycin (aadA1), erythromycin [ere(A)], trimethoprim (dfrA1), and tetracycline [tet(A), tet(B)] were screened using PCR. Our results showed that all swab samples were positive for E. coli. The isolates were uniformly resistant to ampicillin, tetracycline, streptomycin, ciprofloxacin, erythromycin, and trimethoprim-sulphamethoxazole. The isolates exhibited highest susceptibility to colistin sulphate (73.5%), followed by gentamicin (49%), and levofloxacin (17%). All isolates were resistant to three classes of antibiotics, 204 isolates (51%) were resistant to four classes, and 56 isolates (14%) were resistant to five. The highest prevalence of antimicrobial resistance gene was recorded for tetracycline (tet(A):95.25%; tet(B):95.25%) followed by ampicillin (blaTEM:91.25%), streptomycin (aadA1:88.25%), erythromycin (ere(A):84.75%), and trimethoprim (dfrA1:65.5%). In conclusion, surveillance for MDR bacteria in poultry is a critical piece of knowledge, which would be useful for optimizing empiric antimicrobial treatments and exploring alternative antimicrobial agents.

Phasevarions of bacterial pathogens - phase-variable epigenetic regulators evolving from restriction-modification systems.

Phase-variable DNA methyltransferases control the expression of multiple genes via epigenetic mechanisms in a wide variety of bacterial species. These systems are called phasevarions, for phase-variable regulons. Phasevarions regulate genes involved in pathogenesis, host adaptation and antibiotic resistance. Many human-adapted bacterial pathogens contain phasevarions. These include leading causes of morbidity and mortality worldwide, such as non-typeable Haemophilus influenzae, Streptococcus pneumoniae and Neisseria spp. Phase-variable methyltransferases and phasevarions have also been discovered in environmental organisms and veterinary pathogens. The existence of many different examples suggests that phasevarions have evolved multiple times as a contingency strategy in the bacterial domain, controlling phenotypes that are important in adapting to environmental change. Many of the organisms that contain phasevarions have existing or emerging drug resistance. Vaccines may therefore represent the best and most cost-effective tool to prevent disease caused by these organisms. However, many phasevarions also control the expression of current and putative vaccine candidates; variable expression of antigens could lead to immune evasion, meaning that vaccines designed using these targets become ineffective. It is therefore essential to characterize phasevarions in order to determine an organism's stably expressed antigenic repertoire, and rationally design broadly effective vaccines.

An overexpressed cytochrome P450 CYP439A1v3 confers deltamethrin resistance in Laodelphax striatellus Fallén (Hemiptera: Delphacidae).

Deltamethrin resistance in Laodelphax striatellus had been associated with its oxidative detoxification by overexpression of four cytochrome P450 monooxygenases like CYP353D1v2, CYP6FU1, CYP6AY3v2, and CYP439A1v3. The first three P450s have been validated for insecticide-metabolizing capability and only CYP6FU1 was found to degrade deltamethrin. In this study, an investigation was conducted to confirm the capability of CYP439A1v3 to degrade deltamethrin. The CYP439A1v3 was first expressed in Sf9 cell line and its recombinant enzyme was tested for metabolic activity against different insecticides using substrate depletion assay combined with metabolite identification. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and carbon monoxide (CO)-difference spectra analysis showed that the intact cytochrome P450 protein was successfully expressed. Tests with probe substrates proved its enzyme activity, as p-nitroanisole, ethoxycoumarin, and ethoxyresorufin were preferentially metabolized (specific activity 7.767 ± 1.22, 1.325 ± 0.37, and 0.355 ± 0.37 nmol/min per mg of protein, respectively) while only luciferin-HEGE was not. In vitro incubation of the recombinant CYP439A1v3 protein with deltamethrin revealed hydroxylation by producing hydroxydeltamethrin. On the contrary, no metabolite/metabolism was seen with nonpyrethroid insecticide, including imidacloprid, buprofezin, chlorpyrifos, and fipronil. To the best of our knowledge, this is the first study to link a CYP450 from family 439 to confer pyrethroid resistance to L. striatellus. This finding should help in the design of appropriate insecticide resistance management for control of this strain of L. striatellus.

Impact of micronutrient fortification of yoghurt on micronutrient status markers and growth - a randomized double blind controlled trial among school children in Bangladesh.

BACKGROUND: Interventions providing foods fortified with multiple micronutrients can be a cost-effective and sustainable strategy to improve micronutrient status and physical growth of school children. We evaluated the effect of micronutrient-fortified yoghurt on the biochemical status of important micronutrients (iron, zinc, iodine, vitamin A) as well as growth indicators among school children in Bogra district of Bangladesh. METHODS: In a double-masked randomized controlled trial (RCT) conducted in 4 primary schools, 1010 children from classes 1-4 (age 6-9 years) were randomly allocated to receive either micronutrient fortified yoghurt (FY, n = 501) or non-fortified yoghurt (NFY, n = 509). For one year, children were fed with 60 g yoghurt everyday providing 30% RDA for iron, zinc, iodine and vitamin A. Anthropometric measurements and blood/urine samples were collected at base-, mid- and end-line. All children (FY, n = 278, NFY, n = 293) consenting for the end-line blood sample were included in the present analyses. RESULTS: Both groups were comparable at baseline for socio-economic status variables, micronutrient status markers and anthropometry measures. Compliance was similar in both the groups. At baseline 53.4% of the population was anemic; 2.1% was iron deficient (ferritin <15.0 µg/L and TfR > 8.3 mg/L). Children in the FY group showed improvement in Hb (mean difference: 1.5; 95% CI: 0.4-2.5; p = 0.006) as compared to NFY group. Retinol binding protein (mean diff: 0.05; 95% CI: 0.002-0.09; p = 0.04) and iodine levels (mean difference: 39.87; 95% CI: 20.39-59.35; p < 0.001) decreased between base and end-line but the decrease was significantly less in the FY group. Compared to NFY, the FY group had better height gain velocity (mean diff: 0.32; 95% CI: 0.05-0.60; p = 0.02) and height-for-age z-scores (mean diff: 0.18; 95% CI: 0.02-0.33; p = 0.03). There was no difference in weight gain velocity, weight-for-age z-scores or Body Mass Index z-scores. CONCLUSION: In the absence of iron deficiency at baseline the impact on iron status would not be expected to be observed and hence cannot be evaluated. Improved Hb concentrations in the absence of a change in iron status suggest improved utilization of iron possibly due to vitamin A and zinc availability. Fortification improved height gain without affecting weight gain. TRIAL REGISTRATION: ClinicalTrial.gov: NCT00980733.