Propagation of porcine spermatogonial stem cells in serum-free culture conditions using knockout serum replacement.

In vitro culture and expansion of spermatogonial stem cells (SSCs) is an essential prerequisite to enhancing livestock productivity through SSC transplantation. Most of the culture media have been observed to be supplemented with serum. However, the use of serum in culture media may exert detrimental effects on SSC maintenance in vitro. An attempt was made to culture SSCs by replacing serum with 5% 'Knockout Serum Replacement (KSR)' in Doom pig (Sus domesticus), one of the valued indigenous germplasm of North-East India. Testes from 7 to 15 days old piglets were used for isolation, enrichment and in vitro culture of putative SSCs using serum-based and serum-free culture media. The cells were characterized for SSC-specific pluripotent markers expression by immunofluorescence staining and quantitative real-time PCR. The diameter and number of SSC colonies were recorded on days 9, 20 and 30 of culture. Similar morphologies of the SSC colonies were observed in both serum-based and serum-free culture conditions. Colony diameter and colony number were non-significantly higher in serum-free than serum-based media. The cells from both the culture conditions showed high alkaline phosphatase activity. The expression of SSC-specific pluripotent markers was observed in immunofluorescence and quantitative real-time PCR study. The present study revealed that SSCs from porcine species could be maintained in vitro for up to 30 days in serum-free culture using 5% KSR, which is believed to be a promising protein source for improving livestock production, and health care along with their conservation.

Polymorphism and phylogenetic analysis of FecB and FecG genes in native sheep of Meghalaya, India.

The present study was conducted to identify the polymorphism of two major fecundity genes, viz. FecB and FecG, in indigenous sheep of Meghalaya and to assess phylogenetic relationship with 15 breeds of sheep and to estimate the sequence distances between them. Blood samples were collected from 50 adult ewes and PCR-RFLP was performed to detect the polymorphism of these genes. Further digestion of FecB and FecG was done with restriction enzymes AvaII and DdeI, respectively. In the case of FecB gene, two genotypes, viz. AA and AB, were identified where AA genotype yielded one fragment (190 bp) and AB genotype yielded two fragments (160 and 30 bp). The frequencies of A and B alleles were calculated as 0.64 and 0.36 and those of AA and AB genotypes as 0.280 and 0.720 respectively, whereas FecG gene was found to be monomorphic, with only a single genotype designated as AB genotype. Measure of relatedness among Indian and exotic sheep in terms of both the fecundity genes threw light on the evolutionary origin of different sheep breeds.

Allelic variability of estrogen receptor (ESR) gene and its effect on litter traits of Doom pigs.

The present study was conducted to identify the polymorphism of estrogen receptor (ESR) gene and its biological association with litter traits (litter size at birth, litter size at weaning, litter weight at birth and litter weight at weaning) of Doom pigs native to Assam. A total of 50 adult pigs (12 males and 38 females) chosen randomly from three different herds (Herd I, Herd II, and Herd III) were utilized in the present study. Detection of polymorphism of ESR gene was done by means of PCR-RFLP method. The amplified PCR product was digested with Pvu II restriction enzyme. PCR-RFLP analysis of ESR gene revealed polymorphic banding pattern. Two genotypes viz. AA and AB were identified. AA genotype yielded one fragment (120 bp) and AB genotype yielded three fragments (120, 65, and 55 bp). BB genotype was not found in the population under study. The frequencies of A and B alleles were found to be 0.650 and 0.350, respectively, and the genotypic frequencies of ESR gene were found to be 0.300 and 0.700 for AA and AB genotypes, respectively. There was no significant (P > 0.05) effect of ESR genotype on litter traits and the population under study was not in Hardy-Weinberg Equilibrium for ESR gene. Clustal W Multiple alignment of partial sequence of ESR gene revealed single nucleotide changes at 33, 65, 70, 83, and 92nd nucleotide positions. The presence of Pvu II polymorphism and identification of single nucleotide variation of ESR gene opens interesting prospects for improvement of litter traits in Doom pig through selective breeding program, especially based on marker-assisted selection.

Adverse obstetrical and perinatal outcome in adolescent mothers associated with first birth: a hospital-based case-control study in a tertiary care hospital in North-East India.

PURPOSE: To analyze the adverse obstetrical and perinatal outcome of adolescent mothers associated with first birth. PATIENTS AND METHODS: This prospective case-control study was conducted in a tertiary care teaching hospital of North-East India between January 2014 and December 2014. All adolescent primigravidae completing 28 weeks of gestation with singleton pregnancy and delivered at our institution were included in the study group. Primigravidae aged between 20 and 25 years were taken as a control group. Mothers having pregnancy complicated with diabetes mellitus, renal disorder, thyroid disorders, and cardiac diseases were excluded from the study. Demographic data, maternal complications like severe anemia, pre-eclampsia, eclampsia, gestational age at delivery, mode of delivery, and postpartum complications were compared. Among fetal complications, low-birth weight, preterm birth, neonatal intensive care unit admission, still birth, and early neonatal death were compared. All the patients were interviewed regarding contraceptive knowledge and its use preceding the pregnancy. RESULTS: Quality antenatal care was received by 80.6% of adolescent mothers. The adolescent mothers had a higher incidence of pre-eclampsia (odds ratio [OR] 2.017 95% confidence interval [CI]: 1.045-3.894, P=0.03), preterm deliveries (OR: 1.655, 95% CI: 1.039-2.636, P=0.03). Among fetal outcomes, the low- birth weight babies (OR: 1.59, 95% CI: 1.016-2.478), low mean birth weight (2,544.4±622.09 g versus 2,701.6±582.51 g), and higher admission to neonatal intensive care unit (OR: 1.957, 95% CI: 1.120-3.417) were significantly associated with adolescent mothers. There was no significant difference found regarding the mode of delivery, still birth, and early neonatal death. Moreover, contraceptive knowledge and its use were found to be poor among adolescent mothers. CONCLUSION: With quality antenatal, intranatal, and postnatal care, the obstetric risk of childbirth in adolescent mothers may not be as serious as perceived to be.

Necrotizing fasciitis - a rare complication following common obstetric operative procedures: report of two cases.

Necrotizing fasciitis, a near-fatal soft-tissue infection complicating obstetric operative wounds, is a rare entity in obstetrics. Herein, we report two cases of necrotizing fasciitis in severely undernourished and anemic women following obstetric operative procedures. Both undernourishment and anemia compounded the already existing immune-suppressed state in pregnancy and may have lead to life-threatening necrotizing fasciitis. One of the patients developed necrotizing fasciitis following episiotomy and the other following cesarean section. Both the cases were diagnosed clinically. Management was done by total parenteral nutrition, prompt correction of anemia, and surgical debridement under broad-spectrum antibiotic coverage. The raw areas were later reconstructed by split skin grafting in the first case, whereas, in the second case, due to the patient's refusal of skin grafting, the wound was allowed to heal by secondary intention. Both patients survived, although with morbidity. Our study aims to emphasize prompt correction of comorbidities along with aggressive management of necrotizing fasciitis for better outcomes in the obstetric population. Prompt correction of nutritional status improves the survival rate.

Medico-religiousplantsusedbytheHajongcommunityofAssam,India.

ETHNOPHARMACOLOGICAL RELEVANCE: The Hajong people use many medicinal plants in their different rituals. The information generated from the present study regarding the medico-religious plants used by the Hajong tribes need a thorough phytochemical investigation. This could help in creating awareness regarding the need for conservation of such plants and also in the promotion of ethno-medico-botany knowledge within the region besides contributing to the preservation and enrich- ment of the gene bank of such economically important species before they are lost forever. Medico-religious plants so found out in this study should be screened for their efficacy by which new drugs or products could be brought out. There is also need to document this information and compare with that of other tribes in the state and region. MATERIALS AND METHODS: The diseases found during the research work among the Hajong community were grouped in 11 categories. The category of diseases includes plants used for both human and animal diseases. The informants having good knowledge on medico-religious plants were chosen for the interview. Kabiraj or Vaidyas were also included among the informants though they were not available in all villages. The informants were asked about the ritual and religious importance and the medicinal properties of the plants used in different cultural practices. RESULTS: Some 36 plant species including herbs, shrubs and trees are used for curing 51 different diseases with 63 formulations which are recorded during the study. Most of the medicinal plants were collected from their own homestead and rest from the forest. Hajongs make sustainable use of available natural resources that include both medicinal and religious plants. They lead a much disciplined life. They are very much religious and knowledgeable. The survey revealed that not only the Kabiraj but also the elder community member have good knowledge of the medicinal value of some plant usually those species used to treat common diseases like cough, cold, fever, viral fever, headache, stomachache, joint pain, diarrhea, dysentery, minor wounds and cuts etc. Thus 36 plants are found to be used by Hajong in different medico-religious practices. These are regarded as holy plants. These plants are strictly protected by the community. Traditional conservation and management on cultural grounds, therefore, represents a historic contribution to the present day rich biodiversity in the region. CONCLUSION: The whole life system and socio-economic as well as religious and cultural activities of Hajongs were totally dependent upon forests. It reflected their high possession of knowledge on medico religious plants which allowed them to apply their indigenous knowledge regarding various aspects of exploitation of such resources. Different types of important species have been disappeared from the study area, which ultimately creates social, economical and religious problem in Hajong community. The information generated from the present study regarding the medico-religious plants used by the Hajong tribes need a thorough phytochemical investigation. This could help in creating awareness regarding the need for conservation of such plants and also in the promotion of ethno-medico-botany knowledge within the region besides contributing to the preservation and enrichment of the gene bank of such economically important species before they are lost forever. Traditional culture in different Hajong populated areas is very fast declining with lot of traditional knowledge under the influence of dominant culture. Cultural diversity conservation is needed urgently.