This study is conducted with the objective of depicting the relationship between child labour and childhood development. This paper addresses only behavioural, mental, physical development and nutritional problems related to child labour. The study is done on children between 9 and 12 years of age, and children below this age had difficulty in understanding the concepts of the survey questionnaires. Childhood is the critical time for safe and healthy human development. This finding leads strong support to the statement, that poverty is the root cause of child labour. Child labourers are at a high risk of illness and injury, cause due to severe emotional distress, had symptoms of withdrawal and regression, and were prematurely ageing, as well as experiencing depression and low self-esteem. Most of these children are observed to have sleep problems; behavioural problems, heavy consumption of cigarettes, alcohol, infection with sexually transmitted diseases; some criminal activities, unwanted pregnancy and abortion, physical and emotional illness, severe nutritional deficiency and even death due to a wide variety of machinery, biological, physical, chemical, ergonomic and psychosocial hazards as well as from long hours of work and poor living conditions.

The textile industry uses much different kind of fibers as its, raw marital some of this fiber was know and use in the earlier years of civilization, as well as in modern times. Many types of fiber are available some are natural and other are synthetic. Natural fibers that occur in nature can declassified as vegetable, animals and mineral fibers. Silk is most beautiful, lustrous and a perfect fiber, it has been known and used as a textile material for thousands of year. No other fiber was so highly prized by the conquering races of the ancient world. This ancient preference is justified by the latest conclusion of science. Chemical analysis has shown silk to be mainly constitute of fibroin. From soft mucosa substances, fibroin hardens into filament shape and becomes a genius fiber, strong, bright, flexible and elastic. Minutely separable, the fibers of silk, if spun together, make a compact and even thread. These are the characteristic, which the textile worker seeks in every fiber. The ideal is actually found in silk because it is made up of filament so slender, that it can be woven into robes for consumer Because it is so tendentious and flexible, that the thread of silk combine in fabric of impenetrable density, and impervious to light ,moisture or heat. Silk is the protein us extradite ejected by the silk warm.

This study was designed to investigate the impact of buserelin acetate (BA) or human chorionic gonadotropin (hCG) administration on the day of first artificial insemination (AI) on subsequent luteal profile (diameter of corpus luteum (CL) and plasma progesterone) and conception rate in Murrah buffalo. The present experiment was carried out at two locations in 117 buffalo that were oestrus-synchronized using cloprostenol (500 μg) administered (i.m.) 11 days apart followed by AI during standing oestrus. Based on treatment (i.m.) at the time of AI, buffalo were randomly categorized (n = 39 in each group) into control (isotonic saline solution, 5 ml), dAI-BA (buserelin acetate, 20 μg) and dAI-hCG (hCG, 3000 IU) group. Out of these, 14 buffalo of each group were subjected to ovarian ultrasonography on the day of oestrus to monitor the preovulatory follicle and on days 5, 12, 16 and 21 post-ovulation to monitor CL diameter. On the day of each sonography, jugular vein blood samples were collected for the estimation of progesterone concentrations. All the buffalo (n = 117) were confirmed for pregnancy on day 40 post-ovulation. The conception rate was better (p < 0.05) in dAI-BA (51.3%) and dAI-hCG (66.7%) groups as compared to their control counterparts (30.8%). Furthermore, the buffalo of dAI-hCG group had improved (p < 0.05) luteal profile, whereas the buffalo of dAI-BA group failed (p > 0.05) to exhibit stimulatory impact of treatment on luteal profile when compared to control group. In brief, buserelin acetate or hCG treatment on the day of first AI leads to an increase in conception rate; however, an appreciable impact on post-ovulation luteal profile was observed only in hCG-treated Murrah buffalo.

Oxidative stress during peripartum period may compromise the uterine immunity. In the present study, we assessed the oxidative stress and antioxidant status during peripartum period and studied their relationship with postpartum uterine infection in dairy cows. Peripheral blood concentrations of total antioxidant capacity (TAC), malondialdehyde (MDA) and nitric oxide (NO) were determined (day -21, -7, on the day of calving and day +7, +21, +35) in normal (n=11), puerperal metritic (n=7) and clinical endometritic (n=6) cows. Endometrial biopsy was performed on the day of calving and expression of CAT, GPx4 and SOD2 genes was studied using qRT-PCR. Puerperal metritic cows had significantly (P<0.05) lower TAC (on day -7, day 0, day +7, +21 & +35), higher MDA (on day -21, -7 & on the day of calving) and NO (on day 0, +7 & day +35) concentrations compared to normal cows. Similarly, clinical endometritic cows had significantly (P<0.05) lower TAC (on day -7, 0, +7 & +21), higher MDA (on day -21, -7, +7 and +35) and NO (on day +7, +21 & +35) concentrations compared to normal cows. The expression of CAT and GPx4 genes was lower (P<0.05) and SOD2 gene was higher (P<0.05) in endometrial tissue of cows that developed uterine infection compared to normal cows. The relationship of peripheral levels of MDA and NO with antioxidant enzymes expression in endometrial tissue was found significant. Receiver operator characteristic analysis revealed that the concentrations of TAC on day -7 to day +35, MDA on day -21 to day +7 and NO on the day of calving to day +35 were highly correlated to the development of postpartum uterine infection in cows. It may be inferred that the low serum TAC level and high level of lipid peroxidation and NO during peripartum period influenced the endometrial expression of anitioxidative genes that compromised the uterine health during postpartum period.

The experiment was conducted to study ovarian follicular population, its diameter, steroid profile (estradiol 17ß and progesterone) and corpus luteum development during estrous cycle and seasonal anestrous in Murrah buffaloes. They were categorized into 2 groups based on the estrus signs i.e. cyclic (6) and anestrous (6) and subjected to ultrasound scanning of ovaries on day 0 (day of estrus in cyclic and day of first scanning in anestrous buffaloes), 6, 10 and 16 using B-mode scanner equipped with 6.0 MHz linear array transducer. Blood samples were collected on the above days for the estimation of the progesterone and estradiol-17ß. The mean number of small follicles on day 0 and 10 and medium follicles on day 10 were significantly higher within as well as between the cyclic and anestrous buffaloes, while mean number of large follicles on day 0, 10 and 16 were significantly higher in the cyclic as compared to anestrous buffaloes. However, mean number of total follicles were significantly higher in cyclic as compared to anestrous buffaloes on all the days of scanning. The maximum diameter of large follicle was observed in cyclic buffaloes on day 0 (1.40±0.49 cm). The diameter of CL increased with its development and reached to its peak on day 10 (1.39±0.04 cm), thereafter, it reduced on day 16 (0.72±0.30 cm) of the cycle. The mean serum progesterone concentration was significantly higher on day 6, 10 and 16 than day 0 of the estrous cycle indicating development of functional corpus luteum, however, mean serum estradiol-17ß concentration was significantly higher on day 0 than the other days of cycle signifying follicular growth and development secreting higher amount of estrogen.

The objective of this study was to evaluate the effect of cysteamine supplementation on embryo development in mCR2 media. The COCs (1251) were matured in TCM–199 medium containing FSH (5µg/ml), LH (10µg/ml), follicular fluid (10%), FBS (10%) with 3 mg/ml BSA for 27h at 38.5°C and 5% CO2 in an incubator. Matured oocytes were co-cultured with 1×106 spermatozoa/ml collected from a Sirohi buck in fertTALP (10% FBS+ 4mg/ml BSA and 10 µg/ml heparin) for 18 h in incubation. After 18 h of sperm-oocytes, co-incubation of oocytes with sperms were washed in embryo development medium to remove sperm cells adhered to zonapellucida. Presumptive zygotes (1,171) were selected and randomly divided into 2 groups. Group 1, the presumptive zygotes (610) were cultured in mCR2aa as a control for 12 days. Group 2, the presumptive zygotes (561) were culture in mCR2aa medium supplemented with 100 µM cysteamine for 10 days. The percentage of cleavage, morula, and blastocyst production in groups 1 and 2 were 36.39% and 31.71% (cleavage); 21.62% and 30.89% (morula); 4.95% and 8.98% (blastocyst) respectively. In conclusion, results indicated that the addition of cysteamine to the IVC medium stimulates caprine embryo development and blastocyst production.

Porcine circovirus associated diseases (PCVAD) caused by Porcine circovirus type 2 (PCV2) are responsible for heavy losses to the global swine industry in recent decades. PCV2 viral load varies significantly between diseased and subclinically infected pigs. Real time PCR is a highly sensitive technique enabling simultaneous amplification and quantification of specific nucleic acid sequences. Using this technique, the distribution and absolute quantification of PCV2 in naturally infected preweaned piglets was investigated. The viral load in different organs was in the range of (32.92×108 copies/μg DNA to 1.14×108 copies/μg DNA). Results indicate that in PCVAD affected preweaned pigs, lymphoid organs contain the highest viral loads, while intestine and brain the lowest viral loads. This preliminary data on virus load in the lymphoid organs of preweaned piglets will help differentiate mere PCV2 infection and clinical PCVAD.

Thirty six ejaculates from 6 adult Barbari bucks (2-4 years old) maintained at C.I.R.G under semi intensive management system were used to find out the freezability of buck semen at different levels of vitamin C (0.0 mu M, 45.42 mu M, 56.78 mu M, 68.13 mu M) by conventional method of freezing. The ejaculates were collected twice at weekly intervals by artificial vagina. The semen samples were diluted with tris-citric acid fructose diluents having 10% (v/v) egg yolk and 6% (v/v) glycerol as cry protectant agent. The semen samples were extended to maintain sperm concentration approximately 100-120 million per dose. Filling and sealing of straws were done at 5 degrees C in cold handling cabinet after 4h of equilibration period, then straws were vapour frozen for 10 min above 2 cm of liquid nitrogen and finally stored into liquid nitrogen container. Post thaw motility, live sperm count, abnormalities, acrosomal integrity and hypo osmotic swelling test were conducted to check the freezability. Post thaw motility, live sperm count, acrosomal intergrity and hypo osmotic swelling positive spermatozoa differed significantly among groups and they were the highest in 1% group. The result indicated that vitamin Cat the level of 56.78 mu M can be used as an antioxidant in semen diluter in routine freezing process for better post thaw recovery of buck semen. (C) 2015 Elsevier B.V. All rights reserved.

Improvement of reproductive performance of buffaloes requires a better understanding of the mechanisms controlling ovarian follicular growth and development. To study the follicular dynamics and ovarian steroid profile during early pregnancy, 6 buffaloes were selected on the basis of non return rate after insemination and subjected for ultrasonography to confirm the pregnancy as well as follicular development. The mean number of medium, large and total follicles was significantly lower on days 30, 40, 50 and 60 than day 20 of pregnancy indicating reduction in the follicular population with the advancement of pregnancy. There was no significant difference in the diameter of small and medium follicles on different days of pregnancy, however, large follicle significantly decreased with advancement of pregnancy. The progesterone levels fluctuated between 2.52±0.26 and 3.12±0.44 ng/ml until third month of pregnancy. The mean serum estradiol was significantly higher on day 0 and reduced into basal level on first and second month and remained low throughout the study period. In conclusion, on day 20 to 60 of pregnancy, the follicular development is continuous and the number and diameter of larger follicle is decreased as pregnancy advances in buffaloes.

This study was designed to compare the effectiveness of different activation treatments for activation of in vitro matured oocytes and their developmental potency in mCR2aa medium so as to obtain maximum number of embryos. A total of 1090 cumulus oocyte complexes (COC's) were collected from 480 ovaries. In vitro matured oocytes were randomly divided into four groups. Group 1 in vitro matured oocytes (n=226) were exposed to 7% ethanol for 5 min followed by treatment with 2.0 mM DMAP for 4 h in mCR2aa medium. Group 2 in vitro matured oocytes (n=294) were exposed to 7% ethanol for 5 min followed by treatment with 10 µg/ml CHX for 4 h in mCR2aa medium. Group 3 in vitro matured oocytes (n=325) were exposed to 7% ethanol for 5 min followed by treatment with 2.0 mM DMAP and 10 µg/ml CHX for 4 h in mCR2aa medium. Group 4 in vitro matured oocytes (n=108) were cultured for 4 h without any chemical treatment in mCR2aa medium (control). The cleavage rate in groups 1, 2, 3 and 4 was 54.42%, 44.55%, 51.69% and 0.00%, respectively. The percentage of morula and blastocyst production in group 1, group 2 and group 3 was 26.01%, 29.77% and 29.76% and 2.43%, 1.52% and 1.78%, respectively. These results suggest that the activation of in vitro matured oocytes by 7% ethanol for 5 min followed by treatment with 2.0 mM DMAP for 4 h in mCR2aa is most favorable for parthenogenetic caprine embryos production.

The present study aimed to establish the impact of buserelin acetate or hCG administration on day 5 post-ovulation on subsequent luteal profile and conception rate in buffalo. The buffalo (n=45) were subjected to an estrous synchronization protocol (synthetic analog of PGF2α administered, through intramuscular route, 11 days apart), followed by artificial insemination (AI) during mid to late estrus. On day 5 post-ovulation, buffalo were administered (i.m.) normal saline (Control, n=14), buserelin acetate (20μg, d5-BA, n=14) or human chorionic gonadotropin (3000IU, d5-hCG, n=17). Ovarian ultrasonography was conducted on the day of induced estrus and on days 0, 5, 12, 16 and 21 post-ovulation to assess preovulatory follicle or corpus luteum (CL) diameter. Also, on these days, jugular vein blood sampling was conducted for the estimation of plasma progesterone. First service conception rate was greater (χ(2)=5.18, P>0.05) in d5-BA and d5-hCG groups (71.4% and 47.1%, respectively) as compared to control (28.6%). Both treatment groups had a greater (P<0.05) CL diameter and plasma progesterone during the post-treatment period in comparison to that control treatment group. Treatment-induced accessory CL formation was observed in 92.9% and 76.5% buffalo of d5-BA and d5-hCG groups, respectively. In conclusion, buserelin acetate and hCG administration on day 5 post-ovulation leads to accessory CL formation that may have a role in enhancing conception rate.

Ruminant placentas synthesize pregnancy-associated glycoproteins (PAGs) during pregnancy, which serve as biomarkers of pregnancy. The present study was conducted to verify, whether PAGs are expressed in buffalo placenta by using lectin-based affinity chromatography and peptide mass finger printing (PMF). Fetal cotyledonary tissues were collected from gravid uteri procured from slaughtered house. Proteins were extracted and subjected to wheat germ agglutinin (WGA) lectin affinity chromatography to isolate the PAGs. The isolated glycoproteins were separated by one-dimensional SDS-PAGE. PMF results of the 75 kDa protein revealed presence of two PAGs (PAG-7 and -11). The PAG-7 consisted of about 170 mass signals, of which 16 were assigned to corresponding/translated cDNA sequences of buffalo PAG-7, leading to sequence coverage of 40%. PMF result of PAG-11 showed 170 mass signals, of which 15 were assigned to buffalo PAG-11, leading to sequence coverage of 34%. In conclusion, the glycoprotein isolated from placental extract corresponding to 75 kDa band on SDS PAGE gel was a mixture of PAG-7 and -11, which may help in development of suitable diagnostics for pregnancy in buffalo.

ContentsInterferon‐τ (IFN‐τ) has been recognized as the primary embryonic signal responsible for maternal recognition of pregnancy. Uterine endometrium produces both prostaglandin F2α (PGF2α) and prostaglandin E2 (PGE2). PGF2α is responsible for the luteolysis; however, PGE2 favours establishment of pregnancy by its luteoprotective action. In this study, the dose‐response effect of recombinant bovine IFN‐τ (rbIFN‐τ) on prostaglandin (PG) production by buffalo endometrial stromal cells cultured in vitro was studied. Buffalo endometrial stromal cells were isolated by double enzymatic digestion, initially with trypsin III followed by a cocktail of trypsin III, collagenase type II and DNase I and subsequently cultured till confluence. Further, cells were treated with different doses of rbIFN‐τ (0.001, 0.01, 0.1, 1.0 and 10 μg/ml) and keeping a separate set of control. Culture supernatant was collected after 6, 12 and 24 h of treatment. PG levels in the culture supernatant were measured by enzyme immune assay (EIA) and total cellular protein estimated by Bradford method. Results indicated that buffalo endometrial stromal cells following rbIFN‐τ treatment enhanced the secretion of both PGE2 and PGF2α, and also its ratio in a strict dose‐dependent manner with a significant increase (p < 0.01) in PGE2 production at 1 μg/ml dose of rbIFN‐τ and maximal stimulation for both PG was observed at 10 μg/ml. Further, both PG production and its ratio were increased significantly (p < 0.01) in a time‐dependent fashion in all the groups at 6, 12 and 24 h post‐treatment with highest level achieved at 24 h as compared with control. Absolute levels of PGE2 remained higher than PGF2α indicating PGE2 as the major PG produced by endometrial stromal cells. The dose‐dependent response of rbIFN‐τ signifies the importance of optimum concentration of IFN‐τ for the embryonic development especially during the critical period to establish successful pregnancy.

Buffaloes are known for their productivity as compared to average yielding cows due to higher fat percentage, better feed conversion ability and disease resistance. On the other hand, the reproductive performances of buffaloes are often considered as poor owing to late sexual maturity, weak/silent oestrus, repeat breeder and prolonged intercalving interval. The study of cascade of events during oestrus and oestrous cycle can be useful for the improvement of reproductive efficiency of buffaloes. More precisely, the hormonal changes initiated at the molecular level within the animal determine the reproductive nature of the species. Nucleotide/protein sequence analysis serves as a vital tool in analysing the binding of the hormones for their effect or functions. In this study, we have reported cloning and characterization of the complete coding (cDNA) sequence of oxytocin receptor gene (OXTR) in buffaloes. Buffalo OXTR gene contains an uninterrupted ORF of 1176 nucleotides corresponding to an inferred polypeptide length of 391 amino acids (aa). The molecular weight of the deduced aa sequence was found to be 43 kDa with an isoelectric point of 9.253 and 16.328 charge at pH 7.0. The deduced protein sequence consists of 38 strongly basic (+) (K,R), 22 strongly acidic (-) (D,E), 186 hydrophobic (A, I, L, F, W, V) and 95 Polar (N, C, Q, S, T, Y) aa. Results indicated that aspartate (D) at aa position 85 and D, R and C at aa positions 136, 137 and 138, respectively, are conserved in buffaloes. The buffalo OXTR gene shared a per cent similarity ranging from 84.7 to 98.1 and 88.5 to 97.7 at nucleotide and deduced aa sequence levels, respectively, with that of other species. Phylogram constructed on the basis of either nucleotide or deduced aa sequences of buffalo OXTR gene showed that buffalo, cattle and sheep have diverged from human and swine and formed a separate clad. The buffalo sequence has shown maximum similarity and closeness with cattle followed by sheep both at nucleotide and at aa level.

In this study, profiles of glycoproteins of buffalo cotyledons at different stages of pregnancy were studied. First time, WGA lectin affinity chromatography was used to isolate glycoproteins from cotyledons at different stages of gestation in buffalo. The hyper immune sera against the isolated glycoproteins showed no cross reactivity to non- pregnant tissue extract confirming the presence of glycoproteins in samples associated with pregnancy. During early pregnancy, presence of 4 glycoproteins of molecular weights 86, 67, 56 and 51 kDa and out of these glycoproteins, 3 glycoproteins (67, 56 and 51 kDa) were found immunoreactive on western blot analysis. Five glycoproteins of molecular weights 86, 75, 67, 56 and 38 kDa were identified in the mid- and late- stages of pregnancy and all were found immunoreactive. These glycoproteins may be useful for development of RIA/ ELISA based species specific assay for pregnancy diagnosis and monitoring of fetus in buffalo.

The present study was conducted to predict and analyze signaling pathway of buffalo pregnancy associated glycoprotein-1 in silico. Various databases viz. String database, Panther pathways, Biocarta pathways were used for deducing signaling of buffalo pregnancy associated glycoprotein-1. Analysis reveals buffalo pregnancy associated glycoprotein-1 exerts their biological function by interacting with cytokines viz. placenta growth factor and SP1 transcription factor. Placental growth factor belongs to the vascular endothelial growth factor superfamily. Vascular endothelial growth factor is mainly involved in cellular proliferation, migration, differentiation, angiogenesis acting through protein kinase-C signaling pathway. SP1 transcription factor is mostly involved in mediators of transcription and signal transduction during cellular process acting through the SMAD factors. In toto pregnancy associated glycoprotein-1 through these factors may exert its biological functions of angiogenesis, endothelial cell growth, proliferation, migration and differentiation enhancing embryonic growth and development. In conclusion from this study the signaling pathway of buffalo pregnancy associated glycoprotein-1 (PAG-1) was predicted in silico.

Ghrelin, a novel motilin-related endogenous ligand for growth hormone secretagouge receptor, is implicated in various biological functions, including regulation of female reproduction. But the presence of ghrelin and its role in reproductive functions in buffalo, a species with poor reproductive efficiency, is not known. In the present study full-length ghrelin cDNA was isolated from bubaline abomasum, which encodes the entire prepropeptide of 116 amino acids. The deduced amino acid sequence of ghrelin of buffalo showed >95% and 31% identity with that of ruminants (cattle, sheep, and goat) and humans, respectively. Analysis of synonymous and nonsynonymous nucleotide substitutions in the coding region of ghrelin indicated that these sequences of different species have been under purifying selection. The 3995-bp amplicon of ghrelin gene consisting of 4 exons and 3 introns was cloned with genomic DNA from buffalo. Further, ghrelin expression was determined by quantitative real-time PCR, in situ hybridization, and immunohistochemistry in bubaline endometrial tissues at different stages of the estrous cycle and early pregnancy. Our results indicated the persistent expression of ghrelin mRNA and protein in the endometrium during stage I (day 3-5), stage II (day 6-15), and stage III (day 16-21) of the estrous cycle and also during early (similar to day 30-40) pregnancy. Immunohistochemistry and quantitative real-time PCR experiments indicated the relatively higher expression of ghrelin in the endometrium during stage II (day 6-15) of the estrous cycle and early pregnancy than during stage I (day 3-5) and stage III (day 16-21) of the estrous cycle, but no statistically significant difference in ghrelin expression was observed among stages. To conclude, the results of the present study indicate the persistent expression of ghrelin in the uterine endometrium throughout the estrous cycle and in early pregnancy which might be helpful in determining its role in buffalo reproduction. (C) 2013 Elsevier Inc. All rights reserved.

Pregnancy associated glycoprotein-1, a member of the aspartic proteinase family possesses placentogenic and embryo protective functions in numerous domestic species. The present study was conducted to deduce binding and interaction properties of buffalo pregnancy associated glycoprotein-1 (PAG-1) in silico. Buffalo PAG-1 DNA, RNA binding sites deduced using BindN server revealed buffalo PAG-1 protein sequence possess 93 residues with 80% and 56.96% DNA binding specificity and sensitivity, respectively. RNA binding sites are also restricted to identical clusters of amino acid residues as DNA binding sites with 78 residues with RNA binding potential with 80% specificity and 53.95% sensitivity. The potential domains with a high degree of DNA and RNA binding property are present at conserved microsequences residues of buffalo PAG-1 protein sequence. Ligand binding properties from PDBSUM database reveal buffalo PAG-1 possesses ten clefts having potential ligand binding sites consisting of aliphatic and positive amino acid residues. Analysis with the STRING database showed buffalo PAG-1 interaction with cytokines viz. phosphoprotein associated with glycosphingolipid microdomains 1, placenta growth factor precursor, alpha-fetoprotein precursor and SP1 transcription factor. These factors are found to be active during the embryonic stage exerting their functions through angiogenesis, endothelial cell growth, proliferation, migration and differentiation. In conclusion this study reports the various binding properties and putative functional interactions of buffalo pregnancy associated glycoprotein-1 with other cytokines for exerting its biological action.

The objective of the present investigation was to study morphological changes in corpus luteum and luteal progesterone content during different stages of estrous cycle in buffalo (Bubalus bubalis). Uterus along with the ovaries were procured from the local slaughter house and classified into 4 stages; early (stage I, 1 to 5 days), mid (stage II, 6 to 11 days), late luteal phase (stage III, 12 to 16 days) and follicular phase (stage IV, 17 to 20 days) of estrous cycle based on morphological characteristics of corpus luteum and presence of surface follicles on the ovary. Dimensions of the corpus luteum gradually increased with its growth and significant peak values were observed at stage III followed by a decrease at stage IV of estrous cycle, suggesting regression of corpus luteum. The mean progesterone concentration per gram and content per corpus luteum significantly increased from stage I to III of estrous cycle with a peak at stage III and thereafter, it decreased at stage IV. The present study suggested gradual increase in dimension of CL and luteal progesterone contents with the growth of corpus luteum.