Factors influencing somatic cell counts and bacterial contamination in unpasteurized milk obtained from water buffalo in Bangladesh.

Little has been published on the factors influencing the safety and quality of milk derived from water buffalo in Bangladesh. This study aims to describe the milk hygiene parameters and milk chain characteristics of unpasteurized raw milk sold to consumers in order to improve milk hygiene. A quantitative study design evaluated somatic cell counts, total bacterial counts, and specific gram-negative (Enterobacteria) and gram-positive (staphylococci) pathogens in 377 aseptically collected milk samples. Samples were collected at multiple nodes along the buffalo milk value chain: 122 bulk tank milk samples were collected at the farm level, 109 milk samples at the middlemen level, and 111 milk samples at the milk collection centers. In addition, 35 samples were taken from various milk products at the retail level. It was found that progressively increasing somatic cell counts and bacterial counts, including potential pathogens, occurred along the milk chain. A seasonal increase in spring was found, varying based on the farming system (semi-intensive versus intensive). Other factors included water purity and cleanliness of containers, mixing buffalo and cow's milk, and the location of the water buffalo milk producer (coastal or river basin). This study demonstrated how improving udder health and milk hygiene along the water buffalo milk value chain would increase the safety and quality of water buffalo milk in the study area.

Improved recipes provide better complementary feeding in young children: findings of community-based trials in Bangladesh.

BACKGROUND: In Bangladesh, only 34% of children receive minimum acceptable diets as complementary foods (CF) for appropriate age while the rest of them remain at the risk of growth faltering, micronutrient deficiency, compromised cognitive development, and increased morbidity and mortality. OBJECTIVE: The purpose of the study was to develop nutritious recipes for complementary feeding and provide necessary guidelines for improving complementary feeding practices for doctors, nutritionist and caregivers. METHODS: This qualitative study including a baseline survey and Trials of Improved Practices of recipes (TIPs) was conducted on families having 6-23 months old children selected by Multi-stage random sampling methods from 2010 to 2012 throughout seven divisions of Bangladesh. Thirty-five food recipes were developed from local food items based on the 'WHO indicators to assess the adequacy of CF practices' for the TIP. Thirty (30) cooked recipes were analyzed for 10 nutrients in an accredited laboratory in Bangladesh. RESULTS: The baseline survey found that energy density of local recipes was low and did not have the required diversity, adequacy, and nutrient density according to IYCF indicators established by WHO. The TIPs found that 28 recipes out of the 35 recipes were most acceptable by both mothers and children. A manual of improved complementary food recipes and a complementary feeding guideline has been developed with calculation of dietary diversity scores, energy and nutrient density. CONCLUSION: This report will provide evidence-based practical knowledge for training and serve as nutrition education tool for nutrition practitioners, policy makers, and academicians.

Microstructure and Mechanical Property Evaluation of Dune Sand Reactive Powder Concrete Subjected to Hot Air Curing.

The use of different sustainable materials in the manufacture of ultra-high-performance concrete (UHPC) is becoming increasingly common due to the unabating concerns over climate change and sustainability in the construction sector. Reactive powder concrete (RPC) is an UHPC in which traditional coarse aggregates are replaced by fine aggregates. The main purpose of this research is to produce RPC using dune sand and to study its microstructure and mechanical properties under different curing conditions of water curing and hot air curing. The effects of these factors are studied over a long-term period of 90 days. Quartz sand is completely replaced by a blend of crushed and dune sand, and cement is partially replaced by using binary blends of ground granulated blast furnace slag (GGBS) and fly ash (FA), which are used alongside silica fume (SF) to make a ternary supplementary binder system. Microstructural analysis is conducted using scanning electron microscopy (SEM), and engineering properties like compressive strength and flexural strength are studied to evaluate the performance of dune sand RPC. Overall, the results affirm that the production of UHPC is possible with the use of dune sand. The compressive strength of all mixes exceeded 120 MPa after 12 h only of hot air curing (HAC). The SEM results revealed the dense microstructure of RPC. However, goethite-like structures (corrosion products) were spotted at 90 days for all HAC specimens. Additionally, the use of FA accelerated the formation of such products as compared to GGBS. The effect of these products was insignificant from a mechanical point of view. However, additional research is required to determine their effect on the durability of RPC.

Age-Specific Cut-off Values of Amino Acids and Acylcarnitines for Diagnosis of Inborn Errors of Metabolism Using Liquid Chromatography Tandem Mass Spectrometry.

Liquid Chromatography tandem mass spectrometry (LC-MS/MS) is used for the diagnosis of more than 30 inborn errors of metabolisms (IEMs). Accurate and reliable diagnosis of IEMs by quantifying amino acids (AAs) and acylcarnitines (ACs) using LC-MS/MS systems depend on the establishment of age-specific cut-offs of the analytes. This study aimed to (1) determine the age-specific cut-off values of AAs and ACs in Bangladesh and (2) validate the LC-MS/MS method for diagnosis of the patients with IEMs. A total of 570 enrolled healthy participants were divided into 3 age groups, namely, (1) newborns (1-7 days), (2) 8 days-7 years, and (3) 8-17 years, to establish the age-specific cut-offs for AAs and ACs. Also, 273 suspected patients with IEMs were enrolled to evaluate the reliability of the established cut-off values. Quantitation of AAs and ACs was performed on an automated LC-MS/MS system using dried blood spot (DBS) cards. Then the specimens of the enrolled clinically suspected patients were analyzed by the established method. Nine patients came out as screening positive for different IEMs, including two borderline positive cases of medium-chain acyl-CoA dehydrogenase deficiency (MCAD). A second-tier test for confirmation of the screening positive cases was conducted by urinary metabolic profiling using gas chromatography- mass spectrometry (GC-MS). Out of 9 cases that came out as screening positive by LC-MS/MS, seven cases were confirmed by urinary GC-MS analysis including 3 cases with phenylketonuria, 1 with citrullinemia type II, 1 with methylmalonic acidemia, 1 with isovaleric acidemia and 1 with carnitine uptake defect. Two borderline positive cases with MCAD were found negative by urinary GC-MS analysis. In conclusion, along with establishment of a validated LC-MS/MS method for quantitation of AAs and ACs from the DBS cards, the study also demonstrates the presence of predominantly available IEMs in Bangladesh.

Death audit in the neonatal ICU of a tertiary care hospital in Bangladesh: a retrospective chart review.

BACKGROUND: Expectant reduction of neonatal mortality and formulation of preventive strategies can only be achieved by analysis of risk factors in a particular setting. This study aimed to document incidence of neonatal death and to analyze the risk factors associated with neonatal death. METHODS: This retrospective study was carried out in department of Neonatology, Bangabandhu Sheikh Mujib Medical University (BSMMU) over a 12-month period from January to December 2015. The newborns that died within 28 d of life were defined as "Cases" and "Control" were the surviving newborn discharged to home as healthy. Two birth weight and gestational age matched controls were taken for each case. Maternal, obstetric, and newborn characteristics were analyzed between both the groups. Data analysis was performed using SPSS version 20.0 (SPSS Inc., Chicago, IL). A probability of < .05 was considered statistically significant. The strength of association was determined by calculating odds ratio and their 95% confidence intervals (CIs). RESULTS: During the study period, the proportion of death was 9.6% (64/612). Both in Chi-square analysis and in logistic regression analysis, less than four antenatal visits (odds ratio (OR) 2.78; 95% CI: 1.23-6.28, p = .014) and sepsis (OR 2.37; 95% CI: 1.07-5.26, p = .034) were found to be independent risk factors for deaths, whereas LUCS found to be protective for deaths (OR 0.40; 95% CI: 0.19-0.83, p = .015). CONCLUSION: In conclusion, less than four antenatal visits and presence of sepsis were found to be independent risk factors whereas LUCS protective of newborn death.

High resolution melting curve analysis enables rapid and reliable detection of G6PD variants in heterozygous females.

BACKGROUND: Like glucose-6-phosphate dehydrogenase (G6PD) deficient hemizygous males and homozygous females, heterozygous females could also manifest hemolytic crisis, neonatal hyperbilirubinemia or kernicterus upon exposure to oxidative stress induced by certain foods such as fava beans, drugs or infections. Although hemizygous males and homozygous females are easily detected by conventional G6PD enzyme assay method, the heterozygous state could be missed by the conventional methods as the mosaic population of both normal and deficient RBCs circulates in the blood. Thus the present study aimed to apply high resolution melting (HRM) curve analysis approach to see whether HRM could be used as a supplemental approach to increase the chance of detection of G6PD heterozygosity. RESULTS: Sixty-three clinically suspected females were evaluated for G6PD status using both enzyme assay and HRM analysis. Four out of sixty-three participants came out as G6PD deficient by the enzyme assay method, whereas HRM approach could identify nine participants with G6PD variants, one homozygous and eight heterozygous. Although only three out of eight heterozygous samples had G6PD enzyme deficiency, the HRM-based heterozygous G6PD variants detection for the rest of the samples with normal G6PD enzyme activities could have significance because their newborns might fall victim to serious consequences under certain oxidative stress. CONCLUSIONS: In addition to the G6PD enzyme assay, HRM curve analysis could be useful as a supplemental approach for detection of G6PD heterozygosity.

High resolution melting curve analysis targeting the HBB gene mutational hot-spot offers a reliable screening approach for all common as well as most of the rare beta-globin gene mutations in Bangladesh.

BACKGROUND: Bangladesh lies in the global thalassemia belt, which has a defined mutational hot-spot in the beta-globin gene. The high carrier frequencies of beta-thalassemia trait and hemoglobin E-trait in Bangladesh necessitate a reliable DNA-based carrier screening approach that could supplement the use of hematological and electrophoretic indices to overcome the barriers of carrier screening. With this view in mind, the study aimed to establish a high resolution melting (HRM) curve-based rapid and reliable mutation screening method targeting the mutational hot-spot of South Asian and Southeast Asian countries that encompasses exon-1 (c.1 - c.92), intron-1 (c.92 + 1 - c.92 + 130) and a portion of exon-2 (c.93 - c.217) of the HBB gene which harbors more than 95% of mutant alleles responsible for beta-thalassemia in Bangladesh. RESULTS: Our HRM approach could successfully differentiate ten beta-globin gene mutations, namely c.79G > A, c.92 + 5G > C, c.126_129delCTTT, c.27_28insG, c.46delT, c.47G > A, c.92G > C, c.92 + 130G > C, c.126delC and c.135delC in heterozygous states from the wild type alleles, implying the significance of the approach for carrier screening as the first three of these mutations account for ~85% of total mutant alleles in Bangladesh. Moreover, different combinations of compound heterozygous mutations were found to generate melt curves that were distinct from the wild type alleles and from one another. Based on the findings, sixteen reference samples were run in parallel to 41 unknown specimens to perform direct genotyping of the beta-thalassemia specimens using HRM. The HRM-based genotyping of the unknown specimens showed 100% consistency with the sequencing result. CONCLUSIONS: Targeting the mutational hot-spot, the HRM approach could be successfully applied for screening of beta-thalassemia carriers in Bangladesh as well as in other countries of South Asia and Southeast Asia. The approach could be a useful supplement of hematological and electrophortic indices in order to avoid false positive and false negative results.

Molecular Analysis of Glucose-6-Phosphate Dehydrogenase Gene Mutations in Bangladeshi Individuals.

Glucose-6-phosphate dehydrogenase (G6PD) deficiency is a common X-linked human enzyme defect of red blood cells (RBCs). Individuals with this gene defect appear normal until exposed to oxidative stress which induces hemolysis. Consumption of certain foods such as fava beans, legumes; infection with bacteria or virus; and use of certain drugs such as primaquine, sulfa drugs etc. may result in lysis of RBCs in G6PD deficient individuals. The genetic defect that causes G6PD deficiency has been identified mostly as single base missense mutations. One hundred and sixty G6PD gene mutations, which lead to amino acid substitutions, have been described worldwide. The purpose of this study was to detect G6PD gene mutations in hospital-based settings in the local population of Dhaka city, Bangladesh. Qualitative fluorescent spot test and quantitative enzyme activity measurement using RANDOX G6PDH kit were performed for analysis of blood specimens and detection of G6PD-deficient participants. For G6PD-deficient samples, PCR was done with six sets of primers specific for G6PD gene. Automated Sanger sequencing of the PCR products was performed to identify the mutations in the gene. Based on fluorescence spot test and quantitative enzyme assay followed by G6PD gene sequencing, 12 specimens (11 males and one female) among 121 clinically suspected patient-specimens were found to be deficient, suggesting a frequency of 9.9% G6PD deficiency. Sequencing of the G6PD-deficient samples revealed c.C131G substitution (exon-3: Ala44Gly) in six samples, c.G487A substitution (exon-6:Gly163Ser) in five samples and c.G949A substitution (exon-9: Glu317Lys) of coding sequence in one sample. These mutations either affect NADP binding or disrupt protein structure. From the study it appears that Ala44Gly and Gly163Ser are the most common G6PD mutations in Dhaka, Bangladesh. This is the first study of G6PD mutations in Bangladesh.

Antitumor properties of two traditional aromatic rice genotypes (Kalijira and Chinigura).

OBJECTIVE: Methanol extract of bran and unpolished grain of two traditional aromatic rice genotypes viz. Kalijira and Chinigura were assayed for their activity on the growth and initiation of crown-gall tumors on potato disks. MATERIALS AND METHODS: Three Agrobacterium tumefaciens (A. tumefaciens) strain AtSl0105, AtTa0112, and AtAc0114 were used as the tumor forming agent. Collected rice was separated to bran and unpolished grain by different milling processes and made into fine powder before extracting using methanol. Antitumor assay of plant extracts was performed according to standard potato disc bioassay. Disc diffusion assay (Kirby-Bauer Method) was used to screen A. tumefaciens sensitivity test. RESULTS: The results demonstrated a high correlation between the ability of aromatic rice to inhibit the initiation and growth of crown-gall tumors on potato disks. Maximum tumor inhibitions were observed against the strain AtSl0105 by Kalijira bran (73.91%) and Chinigura bran (69.56%). Both unpolished grains showed significant effect (Kalijira 57.43%, Chinigura 55.53%) to inhibit the tumor. CONCLUSION: It can be concluded that aromatic rice (Kalijira and Chinigura) might be a potential source of antitumor agent that can be used for further drug development for tumor treatment.

Maternal and neonatal serum zinc level and its relationship with neural tube defects.

Neural tube defect (NTD) is a multi-factorial disorder in which nutritional, genetic and environmental factors are involved. Among the nutritional factors, low level of serum zinc has been reported from different parts of the world. This hospital-based case-control study was conducted with the objective of finding the relationship between serum zinc level in newborns and their mothers and NTDs in a Bangladeshi population. The study was conducted during August 2006-July 2007 at the Bangabandhu Sheikh Mujib Medical University (BSMMU) in Dhaka. In total, 32 mothers and their newborns with NTDs were included as cases and another 32 mothers with their normal babies were included as controls. Concentration of serum zinc was determined by pyro-coated graphite furnace atomic absorption spectrophotometer (GF-AAS). The mean age of the case and control mothers was 25.28 years and 24.34 years respectively. The mean gestational age of the case newborns was 36.59 weeks and that of the control newborns was 37.75 weeks. The mean serum zinc level of the case and control mothers was 610.2 microg/L and 883.0 microg/L respectively (p < 0.01). The mean serum zinc level of the case and control newborns was 723 microg/L and 1,046 microg/L respectively (p < 0.01). In both case and control groups, the serum zinc level of the newborns positively correlated with that of the mothers. The serum zinc levels of the mothers and newborns negatively correlated with NTDs. Mothers with serum zinc level lower than normal were 7.66 [95% confidence interval (CI) 2.5-23.28] times more likely to have NTDs compared to the normal zinc level of mothers. After adjusting for the zinc level of the newborns, parity, and age of the mothers, this risk reduced 1.61 times [confidence interval (CI) 95% 0.24-8.77]. On the other hand, the low serum zinc level of the newborns was 7.22 times more associated with NTDs compared to the newborns with the normal serum zinc level, which was statistically significant (p = 0.001). After adjusting for other factors, such as maternal age and parity, newborns with the low serum zinc level was found to be 9.186 times more likely to be associated with NTDs compared to newborns with normal serum zinc level. Based on the findings, it may be concluded that the low serum zinc levels of newborns may be associated with NTDs. To confirm these findings, a further study with a larger sample-size is recommended. Moreover, a follow-up study with zinc supplementation to pregnant women and its impact on NTDs is also recommended.

On food and nutrition policy activities in the USA, Australia, and Norway.

Formulation and implementation of a national food and nutrition policy is important for ensuring good health and quality of life. This study examined the formulation and implementation of food and nutrition policies in the USA, Australia, and Norway. Library searches, MEDLINE and POPLINE searches, and personal communications were used for collecting information and data on nutrition activities and policy formulation and implementation in each country. These countries were selected because policy activities have been ongoing since the 1930s with a clear improvement in the nutritional status of the people. Multisectoral participation, conflicts of interest, strategies to alter diet, and attempts to deal with new problems have been highlighted and compared. Findings of the study may be useful to policy-makers in less-developed countries during future policy-making processes.

Effect of deworming on nutritional status of ascaris infested slum children of Dhaka, Bangladesh.

This randomized double blind community trial was conducted on Ascaris infested children (n = 85) aged 2 to 12 years and was aimed to study the impact of deworming on nutritional status, in an urban slum of Dhaka, Bangladesh. The change of weight gain was significantly higher in children given anthelmintics than those given placebo (0.92 kg vs. 0.54 kg). A multiple linear regression model shows that after controlling sex, deworming and height were positively correlated while age and weight were negatively correlated with weight change.