Large-scale Staple Food Fortification as a Complementary Strategy to Address Vitamin and Mineral Vulnerabilities in India: A Critical Review

The slow improvement in micronutrient malnutrition globally and in India warrants a need for scaling?up scientifically proven, cost?effective public health interventions. The present review discusses the potential of staple food fortification as a complementary strategy to tackle micronutrient deficiencies, while addressing the current concerns raised regarding its implementation. The review indicates the below par status of current strategies like dietary diversity and supplementation to address multiple micronutrients deficiencies in India and the need for complementary strategies to tackle this problem. Based on systematic reviews and meta-analysis, global and national evidence has identified staple food fortification as a proven and recognized cost?effective solution to address micronutrient deficiencies. The Government of India has shown a strong leadership to promote this proven intervention. Further, the paper addresses the concern that large?scale staple food fortification (LSFF) may lead to excessive nutrient intakes when delivered together with other interventions, e.g., supplementation, dietary diversity, among the same populations. A key message that emerges from this review is that LSFF is safe with current dietary intake and deficiencies and low coverage of other interventions. Given the current situation of food and nutrition insecurity which the COVID-19 pandemic has further exacerbated, and the critical role that nutrition plays in building immunity, it is even more important that health and nutrition of the population, especially vulnerable age groups, is not only safeguarded but also strengthened. LSFF should be implemented without any further delay to reach the most vulnerable segments of the population to reduce the dietary nutrient gap and prevent micronutrient deficiencies. Effective monitoring and regular dietary surveys will help ensure these interventions are being deployed correctly.

Pre-clinical toxicity & immunobiological evaluation of DNA rabies vaccine & combination rabies vaccine in rhesus monkeys (Macaca mulatta).

Background & objectives: Pre-clinical toxicology evaluation of biotechnology products is a challenge to the toxicologist. The present investigation is an attempt to evaluate the safety profile of the first indigenously developed recombinant DNA anti-rabies vaccine [DRV (100 μg)] and combination rabies vaccine [CRV (100 μg DRV and 1.25 IU of cell culture-derived inactivated rabies virus vaccine)], which are intended for clinical use by intramuscular route in Rhesus monkeys. Methods: As per the regulatory requirements, the study was designed for acute (single dose - 14 days), sub-chronic (repeat dose - 28 days) and chronic (intended clinical dose - 120 days) toxicity tests using three dose levels, viz. therapeutic, average (2x therapeutic dose) and highest dose (10 x therapeutic dose) exposure in monkeys. The selection of the model i.e. monkey was based on affinity and rapid higher antibody response during the efficacy studies. An attempt was made to evaluate all parameters which included physical, physiological, clinical, haematological and histopathological profiles of all target organs, as well as Tiers I, II, III immunotoxicity parameters. Results: In acute toxicity there was no mortality in spite of exposing the monkeys to 10XDRV. In sub chronic and chronic toxicity studies there were no abnormalities in physical, physiological, neurological, clinical parameters, after administration of test compound in intended and 10 times of clinical dosage schedule of DRV and CRV under the experimental conditions. Clinical chemistry, haematology, organ weights and histopathology studies were essentially unremarkable except the presence of residual DNA in femtogram level at site of injection in animal which received 10X DRV in chronic toxicity study. No Observational Adverse Effects Level (NOAEL) of DRV is 1000 ug/dose (10 times of therapeutic dose) if administered on 0, 4, 7, 14, 28th day. Interpretation & conclusions: The information generated by this study not only draws attention to the need for national and international regulatory agencies in formulating guidelines for pre-clinical safety evaluation of biotech products but also facilitates the development of biopharmaceuticals as safe potential therapeutic agents.

Effect of maternal undernutrition on human foetal pancreas morphology in second trimester of pregnancy.

Background & objectives: Maternal undernutrition and hyperglycaemia during pregnancy, as well as foetal undernutrition affecting the development of foetal endocrine pancreas structure and function, especially that of β-cells is well known. This study was undertaken to look into the changes in pancreatic islets morphology of aborted normal human foetuses (16-20 wk old) of undernourished and adequately nourished mothers. Methods: Foetuses were collected over a 24 month period from medically terminated pregnancies of six undernourished mothers (BMI <18.5 kg/m2) and eight adequately nourished mothers (BMI >18.5 kg/m2). The sections were stained with haematoxylin & eosin as well as Masson trichrome for morphometric estimates such as islet count, area, volume, etc. and immunohistochemistry analysis of β-cells for insulin presence was done. Results: Significant correlations between maternal and foetal parameters were seen. However, there were no statistically significant differences in the number, size or density and beta cell counts of the pancreas among foetal pancreas of mothers with BMI <18.5 and >18.5 kg/m2. Interpretation & conclusions: Our findings indicate that nutritional status of the mother may not have profound influence on the morphology of beta cells of foetal pancreas in second trimester of pregnancy. Further studies need to be done to confirm these findings.

Aflatoxin contamination in stored rice variety PAU 201 collected from Punjab, India.

Background & objectives: The present study was carried out on stored rice variety PAU 201 in Punjab that was not permitted for milling and public distribution due to the presence of damaged grains at levels exceeding the regulatory limits of 4.75 per cent. The aim of the study was to determine fungal and aflatoxin contamination in the rice samples to assess hazard from the presence of damaged grains. Presence of iron in discoloured rice grains was also assessed. Methods: Stored samples of paddy of PAU 201 rice variety were collected from six districts of Punjab, milled and analysed for presence of fungal and aflatoxin contamination. Scanning electron microscopy (SEM), energy dispersive X-ray (EDX) analysis and Prussian blue staining was used to determine fungal spores and presence of iron, respectively. Results: Aflatoxin analysis of rice samples indicated that none exceeded the Food Safety and Standards (Contaminants, Toxins and Residues) Regulations, 2011 tolerance limit of 30 μg/kg and majority of the samples had levels <15 μg/kg. The proportion of damaged grains exceeding the limit of 5 per cent was observed in 85.7 per cent of the samples. SEM and Prussian blue staining and EDX analysis of black tipped and pin point damaged rice grains did not show presence of fungal structures and presence of iron. Interpretation & conclusions: The results of the study indicated that the stored rice samples did not pose any health concern with respect to aflatoxin contamination as per the criteria laid down by the Food Safety and Standards Authority of India.

ICMR-DBT Guidelines for Evaluation of Probiotics in Food.

There has been an increased influx of probiotic products in the Indian market during the last decade. However, there has been no systematic approach for evaluation of probiotics in food to ensure their safety and efficacy. An initiative was, therefore, taken by the Indian Council of Medical Research (ICMR) along with the Department of Biotechnology (DBT) to formulate guidelines for regulation of probiotic products in the country. These guidelines define a set of parameters required for a product/strain to be termed as ‘probiotic’. These include identification of the strain, in vitro screening for probiotic characteristics, animal studies to establish safety and in vivo animal and human studies to establish efficacy. The guidelines also include requirements for labeling of the probiotic products with strain specification, viable numbers at the end of shelf life, storage conditions, etc., which would be helpful to the consumers to safeguard their own interest.

Studies on skeletal muscle biopsies in endemic skeletal fluorosis.

Neurological manifestations of skeletal fluorosis have been attributed to compressive radiculomyelopathy. Experimental fluorosis has shown evidence of myopathic changes. Data on human muscle pathology is very scanty. This study included 22 patients with established osteofluorosis. 16 of them showed only EMG changes of neurogenic muscle disease. Histochemistry and histopathology of muscle biopsies showed features of muscle atrophy, evidenced by 'type I' atrophy and 'type I' grouping. No myopathic changes were observed. It may be concluded that the primary changes are related to the nerve, with muscle being affected secondarily. There was no evidence of any primary muscle pathology due to fluorosis.

Nucleolar organizer regions in smooth muscle and breast tumours.

Paraffin sections of formalin fixed tissues, obtained from patients with smooth muscle and breast tumours, were studied by the silver staining technique to quantitate the Nucleolar Organizer Regions (AgNORs) per nucleus and to assess its significance as an independent variable in predicting the behaviour of these neoplasms. Five benign and five malignant tumours of smooth muscle along with ten benign and ten malignant epithelial tumours of breast were studied. Normal myometrium and breast tissue served as controls. Control, benign and malignant tumours of smooth muscle showed mean AgNOR scores of 2.68, 3.89 and 12.50 per nucleus respectively. Control, benign and malignant tumours of breast showed mean AgNOR scores of 1.75, 7.45 and 12.72 per nucleus respectively. These results suggest that quantitative analysis of AgNORs per nucleus is capable of differentiating benign from malignant lesions of smooth muscle and breast.

Effects of ovulen-50, diethylnitrosamine and phenobarbital on liver regeneration in female rats.

Short term effects of ovulen-50, a combination type oral contraceptive agent and phenobarbital—an established hepatic tumour promoter, were examined in the livers of diethylnitrosamine-initiated and uninitiated female rats. Liver mitotic activity as judged by liver weight, [3H] thymidine incorporation into DNA and levels of DNA, RNA and protein were measured in non-regenerating and regenerating liver. Hepatic γ-glutamyl transpeptidase activity and hepatocyte agglutination with concanavalin A were examined in diethylnitrosamine- and/or phenobarbital-treated rats. The results indicate that diethylnitrosamine or ovulen-50 individually are mitoinhibitory in regenerating liver. Phenobarbital alone has a slight mitostimulatory effects in nonregenerating liver, but no effect on liver regeneration. Administration of ovulen-50 and phenobarbital to diethylnitrosamine initiated rats mitigated the mitoinhibition during regeneration. Contrary to the earlier observation with ovulen-50, neither phenobarbital nor diethylnitrosamine induced hepatocyte agglutination in the presence of concanavalin A. Like ovulen-50, diethylnitrosamine also increased the level of hepatic γ-glutamyl transpeptidase. Phenobarbital produced only insignificant rise and did not substantially exacerbate the effect diethylnitrosamine. The data show that though some of the effects of ovulen-50 resemble those of diethylnitrosamine or phenobarbital, the changes observed may not be related to the neoplastic phenomenon since they were not seen in an initiator-promoter combination regimen.