Evidence of prenatal toxicity of herbal based indigenous formulations for sex selection in rat models.

Indigenous preparations(IPs) for a male child is reported from some parts of India. The present study aims to explore the effects of IPs for sex selection or sex selection drugs (SSDs) on pregnancy outcomes in rat models. SSDs contain Bryonia laciniosa, Quercus infectoria and Putranjiva roxburghii along with other ingredients. METHODS: An experimental design with successfully mated female rats were randomized into control and treatment groups. Phase 1 had 2 interventional arms while phase 2 had 3 interventional arms (12 rats/arm) besides control arm. In phase-1, pregnant females were dosed two SSDs(1000 mg/kg) on gestation days 1-5 whereas, in phase-2, on gestation days 6-19 to correlate the effect of the SSDs (500/1000/1500 mg/kg) consumption during different stages of pregnancy. Pregnant females were observed for clinical signs following treatment. The rats were sacrificed one day before expected day of delivery for evaluation. Pregnancy rate, gestation index, number of corpora lutea, and litter size were assessed. Foetuses were examined for sex, skeletal and soft tissue alterations. DISCUSSION AND CONCLUSION: In phase 1, no appreciable findings were there with SSD exposure. In phase 2, intrauterine growth and survival of foetuses were affected when SSDs were administered during organogenesis period. Decreased number of live foetuses and increased incidence of early and late resorption, reduced fetal growth with significant alteration in skeleton and viscera were found in treatment groups in a dose-dependent manner. This correlates well with findings from observational studies in pregnant women. However, such treatment at any dose did not effect sex differentiation.

Heavy Metals in Indigenous Preparations Used for Sex Selection During Pregnancy in India.

Indigenous preparations (IPs) have evoked a considerable interest in alleviating infections and chronic diseases and improving wellbeing. While such formulations have been a part of traditional practice in several countries and many have been reviewed scientifically for their claims, several of them until date remain to be investigated. A class of IPs for sex selection by Indian pregnant women exists with an aim of begetting a male offspring. In view of the leads obtained from our previous studies on detrimental effects of the newborn, for instance stillbirths and congenital malformations, we attempted to investigate the samples for heavy metal toxicity. Three samples were chosen following phytochemical analysis and reproductive toxicity of such preparations under in vivo conditions. The selected samples were examined for heavy metals-lead, cadmium, arsenic, and mercury using Microwave-assisted atomic absorption spectroscopy. The upper limit level of lead, mercury, and cadmium was found to be 18.56, 0.11, and 0.84 mg/kg respectively whereas arsenic was not detected. The levels of lead and mercury were found to be manifolds high in the IP samples that were primarily contributed by its constituents. The results of our study indicate the potential risk conferred upon, to both the mother and fetus on account of high levels of lead, mercury, and cadmium.

Global sex selection techniques for family planning: a narrative review.

OBJECTIVE: To document the varying methods of sex selection, both primitive (traditional) as well as advanced forms available around the world. CONTEXT: With the increasing desire of couples to choose the gender of their offspring, scientific sex-selection methods and techniques have evolved over time; unfortunately, the medical and social consequences have remained poorly emphasised. METHODS: We searched electronic search engines and grey literature that included research articles from journals, books, websites and news articles in English until August 2016. We comprehensively compiled the findings such as underlying principles, time of use in relation to conception and others. RESULTS: We classified the techniques into natural methods that rely on physiological conditions and artificial methods, including manipulation of seminal fluid for sex selection. Natural methods include Shettles technique, Whelan Method, Billings Ovulation Method, pre-conception diet, and gender selection kits such as GenSelect and Smart Stork, which rely on timing of intercourse, the vaginal environment, a selective diet and nutraceuticals. More advanced and artificial methods include sperm sorting or Ericsson's method, Microsort, Preimplantation Genetic Diagnosis and Urobiologics PreGender test. The markets for these techniques are prevalent in India where the birth of a son is desired. There is also widespread use of indigenous medicines for sex selection. The review reports side effects such as vaginal infections, hyperstimulation syndrome, multiple pregnancies, birth defects and stillbirths. CONCLUSION: We conclude that sex-selection practices need urgent intervention in view of the social harm, unwarranted gender bias, and diversion of resources from genuine medical need.

Lactobacillus casei stimulates phase-II detoxification system and rescues malathion-induced physiological impairments in Caenorhabditis elegans.

Malathion, an organophosphorus insecticide, is renowned for its inhibitory action on acetylcholinesterase (AChE) enzyme that eventually leads to widespread disturbance in the normal physiological and behavioral activities of any organism. Lactic acid bacteria (LAB) are still an underexploited and inexhaustible source of significant pharmaceutical thrust. In the present study, Caenorhabditis elegans was employed to identify and characterize the indigenous LAB isolated from different traditional food against malathion-induced toxicity. The results demonstrated that malathion at its LD50 concentration decreased various C. elegans physiological parameters such as survival, feeding, and locomotion. Among the screened isolates, L. casei exhibited an excellent protective efficacy against malathion-induced toxicity by increasing the level of AChE and thereby rescued all physiological parameters of C. elegans. In addition, short-term exposure and food choice assay divulged that L. casei could serve as a better food to protect C. elegans from noxious environment. The expression analysis unveiled that L. casei gavage upregulated the phase-II detoxification enzymes coding genes metallothioneins (mtl-1 and mtl-2) and glutathione-S-transferase (gst-8) and thereby eliminated malathion from the host system. Furthermore, the upregulation of ace-3 along with down-regulation of cyp35a in the nematodes supplemented with L. casei could be attributed to attenuate the malathion-induced physiological defects in C. elegans. Thus, the present study reports that an indigenous LAB-L. casei could serve as a promising protective agent against the harmful effects of pesticide.

Consumption of indigenous medicines by pregnant women in North India for selecting sex of the foetus: what can it lead to?

BACKGROUND: Sex ratio is an important indicator of development. Despite all the measures undertaken for improvement, it remains an issue of concern in India, with Haryana having a very low sex ratio in the country. Studies have been conducted indicating that consumption of indigenous drugs used for sex selection (SSD) could be strongly associated with adverse effects on the foetal development, including congenital malformations. Some samples of SSDs were collected from parts of North India and analysed in a standard laboratory for its components. METHODS: Thirty SSDs used by the local community were procured from various sources in north India through a rigorous process of collection. These were subjected to laboratory tests to investigate the presence of phytoestrogen and testosterone. Following sample extraction, thin layer chromatography and high performance liquid chromatography were carried out for analysing phytoestrogen content. RESULTS: SSDs were available in various forms such as powder, tablets, mostly from faith healers. Around 87% of the samples collected from sources like doctors, quacks and faith healers were to be taken by the pregnant women after conception; 63% drugs were strongly positive for phytoestrogens (genistein, daidzein, formononetin) and 20% drugs were positive for testosterone. The average dose of the components as calculated after analyses was as follows: daidzein--14.1 mg/g sample, genistein--8.6 mg/g sample, formononetin--5 mg/g sample. CONCLUSION: These SSDs could be potentially detrimental to the growth and development of the foetus. This is likely to have implications on the health of the community. In view of the results obtained in our study, we strongly attest the importance in curbing this harmful practice by banning the supply of the drugs as well as by advocating behavioural changes in the community.

Defect detection around rebars in concrete using focused ultrasound and reverse time migration.

Experimental and numerical investigations have been performed to assess the feasibility of damage detection around rebars in concrete using focused ultrasound and a Reverse Time Migration (RTM) based subsurface imaging algorithm. Since concrete is heterogeneous, an unfocused ultrasonic field will be randomly scattered by the aggregates, thereby masking information about damage(s). A focused ultrasonic field, on the other hand, increases the possibility of detection of an anomaly due to enhanced amplitude of the incident field in the focal region. Further, the RTM based reconstruction using scattered focused field data is capable of creating clear images of the inspected region of interest. Since scattering of a focused field by a damaged rebar differs qualitatively from that of an undamaged rebar, distinct images of damaged and undamaged situations are obtained in the RTM generated images. This is demonstrated with both numerical and experimental investigations. The total scattered field, acquired on the surface of the concrete medium, is used as input for the RTM algorithm to generate the subsurface image that helps to identify the damage. The proposed technique, therefore, has some advantage since knowledge about the undamaged scenario for the concrete medium is not necessary to assess its integrity.

Indigenous Medicine Use for Sex Selection During Pregnancy and Risk of Congenital Malformations: A Population-Based Case-Control Study in Haryana, India.

INTRODUCTION: Congenital malformations (CMFs) are a major public health problem in India. Consanguineous marriages, infections during pregnancy, folic acid deficiency during the periconceptional period, exposure to pesticides and a history of intake of drugs during pregnancy have been hypothesized as risk factors. Drugs include oral contraceptive pills, progesterone analogues, medications for ailments and indigenous drugs to bear male offspring. It is important to analyze the risk factors in order to implement preventive measures. The prime objective of this study was to study the risk factors of visible structural CMFs, with a focus on indigenous medicines for sex selection. METHODS: A population-based, case-control study was undertaken in Haryana state. Cases included children (0-18 months) with any apparent structural deformity as reported by various Government sources. A consecutive birth from the same area as the case was labelled and included as the control. The sample size calculated was 175 in each group. Mothers of every case and control were interviewed at their respective homes using a structured tool. Descriptive analysis, bivariate analysis, followed by logistic regression was conducted to establish the association between risk factors and CMFs. RESULTS: The sociodemographic profiles of the cases and controls were similar. Among the various risk factors studied, more than two living children (unadjusted odds ratio [OR] 1.6, 95 % CI 1.04-2.4) and intake of sex-selection drugs (unadjusted OR 2.8, 95 % CI 1.6-5.1) were significant risk factors on bivariate and regression analyses. The risk of having a child with CMFs was threefold more among mothers with a history of intake of indigenous medicines for sex selection (adjusted OR 3; 95 % CI 1.7-5.6). CONCLUSIONS: The intake of indigenous drugs during pregnancy increased the risk of CMFs almost threefold. This has social as well as economic implications, and hence needs further investigation.

Antimicrobial efficacy and in vivo toxicity studies of a quaternized biopolymeric flocculant.

This study evaluated the antibacterial spectrum and safety of a chemically modified biopolymeric flocculant (TMB) against waterborne pathogens. The biopolymer previously characterized as polysaccharide with flocculating activity is produced extracellularly by the bacterium Klebsiella terrigena. The amino sugars on the polymer were chemically modified by quaternization, which resulted in N,N,N trimethyl biopolymer (TMB). Quaternization was effective in imparting biocidal activity to TMB against five selected waterborne pathogens, namely, Aeromonas hydrophila, Yersinia enterocolitica, Salmonella typhimurium, Listeria monocytogenes and Escherichia coli O157:H7. 99.999% inactivation was achieved with S. typhimurium at a dose of 60 µg ml(-1) of TMB within 60 min at the ambient temperature, followed by other pathogens. Haemotological, histopathological and general examinations indicated no adverse effects in Swiss albino mice fed with the quaternized biopolymer (120 mg kg(-1) body weight(-1) day(-1)) over a period of 30 days. These results suggested that TMB was tolerated well without any signs of toxicity and may have potential application as a safe, antimicrobial bioflocculant for both removing and inactivating waterborne pathogens.

Novel synergistic approach to exploit the bactericidal efficacy of commercial disinfectants on the biofilms of Salmonella enterica serovar Typhimurium.

Combined effect of malic acid and ozone as sanitizer to inhibit the biofilm formation by Salmonella typhimurium on different food contact surfaces was investigated in this study. Different surfaces used in food industry including PVC pipes, polyethylene bags, plastic surfaces and fresh produce were analyzed for the biofilm formation by S. typhimurium ST1 and ST2. Malic acid alone was not able to inhibit biofilm formation in all the samples. However, combination of malic acid with ozone reduced the biofilm formation on plastic bags as well as on PVC pipes suggesting as an effective disinfectant for food contact surfaces. Five- and six-fold reduction in biofilm formation was observed in microtitre plates after 20 h and 40 h. Scanning electron micrographs of carrot and turnip showed control over the biofilms. Malic acid as sanitizer in food industry was effective for the complete inhibition of biofilm in carrot and other food contact surfaces, besides this, combined sanitizer (malic acid and ozone) was effective in turnip. Biofilms in food-processing industries can survive even after the sanitizer treatment and may represent reservoirs of product contamination leading to subsequent spoilage and/or food safety risks.

Probiotic characterization of potential hydrolases producing Lactococcus lactis subsp. lactis isolated from pickled yam.

The aim of this study was to characterize potential probiotic strain co-producing α-amylase and ß-galactosidase. Sixty-three strains, isolated from pickle samples were screened for their hydrolase producing capacity by utilizing different starches as carbon source. One out of 63 strains, isolated from traditionally fermented pickled yam showing maximum hydrolase activity (α-amylase (36.9 U/ml) and ß-galactosidase (42.6 U/ml)) within a period of 48 hours was identified as Lactococcus lactis subsp. lactis. Further, it was assessed for the probiotic characteristics under gastrointestinal conditions like acidic, alkaline, proteolytic enzymes, bile stress and found to exhibit tolerance to these stresses. The therapeutic potential of the isolate is implicated because of its antagonistic effect against enteric foodborne pathogens (Salmonella typhimurium, Escherichia coli 0157:H7, Staphylococcus aureus, Yersinia enterocolitica and Aeromonas hydrophila). The results of this study entail a potential applicability of the isolate in developing future probiotic foods besides the production of industrially significant hydrolases.

Development of exobiopolymer-based biosensor for detection of phosphate in water.

The present study was conducted to develop a biosensor by exploiting phosphate-binding capacity of exobiopolymer (EBP) produced by Acinetobacter sp. An environmental isolate of EBP-producing Acinetobacter sp. was subjected to transposon (Tn5) mutagenesis to overproduce EBP and afford improved phosphate selectivity. A mutant producing the highest amount of EBP with high phosphate-binding capacity was selected for biosensor probe fabrication. Phosphate samples were filtered through EBP-coated membranes and phosphate retained on membranes was determined by molybdenum blue method. The color produced was read using a LED 690 nm/photodiode detection system linked to an amplifier and signals were converted to appropriate phosphate concentrations. The biosensor had a limit of detection of 0.5 mg/L and a limit of quantification 1 mg/L. The biosensor as well as the probe were found to be stable for at least 28 days. In conclusion, we believe that the biosensor may have applications in monitoring of wastewater and environmental samples. Further, the enrichment of phosphate levels by EBP can help in analysis of very low phosphate concentrations.

Interpretation of Indian orthodox tea using multi-sensing impedometric technique.

Twelve different orthodox Indian tea samples were collected from tea growing and processing sites of Kangra (Valley), India. The percentage of major chemical constituents responsible for the tea quality has been determined by HPLC and UV-vis spectrophotometer. Impedance response using Platinum (Pt), Gold (Au), Silver (Ag), Glassy Carbon (GC), Polyaniline (PANI) (emraldine salt) and Poly Pyrrole (PPY) working electrodes in tea infusions in the frequency range of 1 Hz to 100 kHz has been measured for 30 days on each tea sample. The impedance response of these working electrodes along with the determined chemical concentrations was subjected to Principal Component Analysis (PCA). It was possible to map the antioxidant levels in these tea samples with few exceptions from the score plot of PCA.

Modeling in vitro cholesterol reduction in relation to growth of probiotic Lactobacillus casei.

Lactobacillus casei LA-1 isolated from a nondairy fermented source was evaluated for its in vitro ability to reduce cholesterol. The bacterium tested positive for bile salt deconjugation in relation to cholesterol removal. Tested growth-associated physiological variables such as pH, temperature and inoculum size were all found to have significant effects on in vitro cholesterol reduction and biomass production (both P < 0.005). Furthermore, a central composite design was used to evaluate the effects of significant variables and their interactions. A linear regression model was developed for in vitro cholesterol reduction as a function of growth-associated variables. Maximum cholesterol reduction achieved was 45% whereas maximum biomass yield of 2.34 optical density was observed at the central point. Our study possibly indicates that the growth of L. casei LA-1 depends on its cholesterol removing ability.

Use of Lactococcus lactis to enrich sourdough bread with γ-aminobutyric acid.

Fried sourdough bread (bhatura) with an elevated amount of γ-aminobutyric acid (GABA) was produced using lactic acid bacteria (LAB). The LAB starter was screened and isolated from pickled yam showing highest GABA content and was identified as Lactococcus lactis subsp. lactis. The maximum GABA production in de Man Rogosa Sharpe (MRS) media supplemented with monosodium glutamate (MSG) was 110 mg/100 ml at pH 5, and 1-3% NaCl did not change the production of GABA significantly (p>0.05). When MSG was replaced with Vigna mungo in sourdough, the amount of GABA for bhatura was 226.22 mg/100 g representing about 10-fold increase. A sensory evaluation resulted as the overall general acceptability of bhatura to be 4.91 ± 0.03 on a five-point hedonic scale. Thus, the results indicated the potential of L. lactis as a LAB starter for the production of GABA-enriched bhatura. Although other physiological effects can be expected in the product, animal and clinical studies are mandatory prior to application of this food.

Impedance study of tea with added taste compounds using conducting polymer and metal electrodes.

In this study the sensing capabilities of a combination of metals and conducting polymer sensing/working electrodes for tea liquor prepared by addition of different compounds using an impedance mode in frequency range 1 Hz-100 KHz at 0.1 V potential has been carried out. Classification of six different tea liquor samples made by dissolving various compounds (black tea liquor + raw milk from milkman), (black tea liquor + sweetened clove syrup), (black tea liquor + sweetened ginger syrup), (black tea liquor + sweetened cardamom syrup), (black tea liquor + sweet chocolate syrup) and (black tea liquor + vanilla flavoured milk without sugar) using six different working electrodes in a multi electrode setup has been studied using impedance and further its PCA has been carried out. Working electrodes of Platinum (Pt), Gold (Au), Silver (Ag), Glassy Carbon (GC) and conducting polymer electrodes of Polyaniline (PANI) and Polypyrrole (PPY) grown on an ITO surface potentiostatically have been deployed in a three electrode set up. The impedance response of these tea liquor samples using number of working electrodes shows a decrease in the real and imaginary impedance values presented on nyquist plots depending upon the nature of the electrode and amount of dissolved salts present in compounds added to tea liquor/solution. The different sensing surfaces allowed a high cross-selectivity in response to the same analyte. From Principal Component Analysis (PCA) plots it was possible to classify tea liquor in 3-4 classes using conducting polymer electrodes; however tea liquors were well separated from the PCA plots employing the impedance data of both conducting polymer and metal electrodes.

Mitogenic response and probiotic characteristics of lactic acid bacteria isolated from indigenously pickled vegetables and fermented beverages.

Lactic acid bacteria from indigenous pickled vegetables and fermented beverages (fermented rice and Madhuca longifolia flowers) were isolated and investigated for their functional characteristics in vitro as potential new probiotic strains. Four isolates (all Lactobacillus spp.) selected on the basis of high tolerance to bile (0.2%) were identified by standard and molecular methods (16S rDNA) as L. helveticus, L. casei, L. delbrueckii and L. bulgaricus from pickled vegetables and fermented beverages respectively. These selected strains had antibiotic resistance, tolerance to artificial gastric juice and phenol (0.4%), enzymatic profile, and antagonistic activity against enteric pathogens (Enterobacter sakazakii, Salmonella typhimurium, Shigella flexneri 2a, Listeria monocytogenes, Yersinia enterocolitica and Aeromonas hydrophila). All strains survived well in artificial gastric juice at low pH (3.0) values for 4 h, possessed bile salt hydrolase activity and were susceptible to most antibiotics including vancomycin. Additionally, the isolates exhibited high tolerance to phenol, high cell surface hydrophobicity (>60%) and induced proliferation of murine splenocytes. All the four strains of present study suppressed the Con A-stimulated proliferation of the mouse spleen cells, although L. casei had the strongest suppressive effect. The results of this study suggest a potential application of the strains (following human clinical trials), for developing probiotic foods.

Modeling of permeabilization process in Pseudomonas putida G7 for enhanced limonin bioconversion.

A facile process of enhanced whole cell biotransformation to debitter the triterpenoid limonin in citrus juices was optimized in this work. To maximize bioconversion, permeabilization conditions were modeled using response surface methodology. A central composite rotatable design with four significant variables (concentration, temperature, pH, and treatment time) was employed. The second order polynomial equations with R² values above 0.9 showed good correspondence between experimental and predicted values. The concentration, temperature, pH, and treatment time as well as their interactions had significant effects (p < 0.001) on limonin bioconversion. The optimum operating conditions for permeabilization were observed at a Na2EDTA concentration of 1.5 µM, treatment time of 15 min, temperature of 28 °C, and pH 8. A maximum reduction of 76.71% in the limonin content was achieved within 150 min under selected conditions. The results are promising for refining permeabilization technique for whole cell biocatalysts thereby improving the debittering of citrus juices significantly.

Enhancement of bioconversion efficiency of limonin by Pseudmonas putida G7.

The biocatalytic activity of periplasmic dehydrogenase of Pseudomonas putida G7 strain to catalyse limonin was enhanced when whole cells were permeabilized with EDTA (1 µM) lysozyme (100 µg/ml). The treated cells were entrapped in dialysis membranes to increase the stability. Permeabilized cells (1 g dry weight) entrapped in dialysis membrane could biotransform 73.67% of limonin in unpasteurized mandarin juices in a single-batch cycle of 3 h. Furthermore, permeabilized cells stored for 45 days in phosphate buffered saline (at 4 or 30°C) retained enzyme activity and were reusable for up to eight batch cycles of limonin reduction. The results of this study suggest a potential application of permeabilized P. putida G7 cells for reducing limonin levels in mandarin juices.

Industrial Whey Utilization as a Medium Supplement for Biphasic Growth and Bacteriocin Production by Probiotic Lactobacillus casei LA-1.

The ability of probiotic Lactobacillus casei LA-1 for bacteriocin production using industrial by-products, such as whey, as supplement in growth medium has been demonstrated for the first time. Whey was investigated as a sole carbon source in cooperation with other components to substitute expensive nutrients as MRS for economical bacteriocin production. Industrial whey-supplemented MRS medium was then selected as to determine the effect of four variables (temperature, initial pH, incubation time, and whey concentration) by response surface methodology on bacteriocin production. Statistical analysis of results showed that two variables have a significant effect on bacteriocin production. Response surface data showed maximum bacteriocin production of 6,132.33 AU/mL at an initial pH of 7.12, temperature 34.29 °C, and whey concentration 13.74 g/L. The production of bacteriocin started during the exponential growth phase, reaching maximum values at stationary phase, and a biphasic growth and production pattern was observed. Our current work demonstrates that this approach of utilization of whey as substitution in costly medium as MRS has great promise for cost reduction in industry for the production of novel biological metabolic product that can be utilized as a food preservative.