Cryopreservation of Oocytes and Embryos: Current Status and Opportunities

The biochemical and metabolic activities of living cells are virtually stopped at ultralow temperature and they enter into a suspended state of animation. However, as such, exposure of living cells to ultralow temperature is associated with complex changes that reduce their survivability following freeze-thawing. Cryopreservation is the method for preserving living cells at ultralow temperature at genetically and physiologically stabilized state. Cryopreservation of oocytes and embryos is an integral part of the assisted reproductive technologies with many potential applications. Cryobanking of oocytes and embryos derived from genetically superior animals is promising for enhancing the outcome of planned breeding programs and conserving biodiversity of endangered animal species. Cryobanking can also ensure steady supply of oocytes and embryos for many downstream applications of assisted reproduction such as in vitro embryo production, embryo transfer, production of stem cells, and genetic engineering. Tremendous advancements have been made in this field over the past 5 decades and several methods have been demonstrated for cryopreserving oocytes and embryos in different species. This chapter focuses on the fundamental aspects of oocyte and embryo cryopreservation

GHG Emissions from Livestock: Challenges and Ameliorative Measures to Counter Adversity

Livestock and climate change are interlinked through a complex mechanism and serve the role of both contributor as well as sufferer. The livestock sector is primarily accountable for the emission of methane and nitrous oxide. Methane emission takes place from both enteric fermentation and manure management; whilst nitrous oxide emission is purely from manure management. Rumen methanogenesis due to emission intensity and loss of biological energy always remains a priority for the researchers. Greenhouse gas (GHG) emissions from manure are determined by storage conditions and the organic content of the manure waste. Due to large livestock population, India is a major contributor of enteric methane emission, while its contribution to the excrement methane is negligible. In this chapter, information pertaining to enteric methane emission, excrement methane and nitrous oxide emissions and ameliorative/precautionary measures for reducing the intensity of emissions have been compiled and presented

IL1B Induced Smad 7 Negatively Regulates Gastrin Expression

BACKGROUND: Helicobacter pylori elicited IL1B is one of the various modulators responsible for perturbation of acid secretion in gut. We have earlier reported that IL1B activated NFkB downregulates gastrin, a major modulator of acid secretion. However, we hypothesized that regulation of gastrin by IL1B would depend on the cell's ability to integrate inputs from multiple signaling pathways to generate appropriate biological response. PRINCIPAL FINDING: In this study, we report that IL1B induces Smad 7 expression by about 4.5 fold in gastric carcinoma cell line, AGS. Smad 7 resulted in transcriptional repression of gastrin promoter by about 6.5 fold when co-transfected with Smad 7 expression vector and gastrin-promoter luciferase in AGS cells. IL1B inhibited phosphorylation of Smad 3 and subsequently interfered with nuclear translocation of the positive Smad complex, thus occluding it off the gastrin promoter. IL1B promoter polymorphisms (-511T/-31C IL1B) are known to be associated with H. pylori associated gastro-duodenal ulcer. We observed that IL1B expressed from -31T promoter driven IL1B cDNA elicited 3.5 fold more Smad 7 than that expressed from the IL1B-31C variant in AGS cells. This differential activation of Smad 7 by IL1B promoter variants translated into differential downregulation of gastrin expression. We further analyzed Smad 7, NFkB, IL1B and gastrin expression in antral gut biopsy samples of patients with H. pylori associated duodenal ulcer and normal individuals. We observed that individuals with duodenal ulcer had significantly lower levels of IL1B, Smad 7, NFkB and corresponding higher level of gastrin expression. CONCLUSION: Pro-inflammatory cytokine IL1B repress gastrin expression by activating Smad 7 and subsequent inhibition of nuclear localization of Smad 3/4 complex. Polymorphic promoter variants of IL1B gene can modulate the IL1B expression which resulted in differential activation Smad 7 and consequent repression of gastrin expression, respectively. Analysis of H. pylori infected duodenal ulcer patient's gut biopsy samples also supported this observation

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Not AvailableThe post-thaw viability and motility of buffalo sperm are reduced significantly when they are cryopreserved. One of the etiologies of this reduced post-thaw viability and motility of buffalo sperm has been attributed to cryopreservation-associated generation of reactive oxygen species (ROS) in sperm. Hence, there is an urgent need to address this issue by incorporating some of the external additives which can reduce or scavenge the production of ROS. Sperm were separated from seminal plasma as it contains many identified and unidentified motility stimulating/inhibiting factors that may interfere in the interpretation of any action of external agent. Thus, in the present study, sperm were cultured in vitro in sp-TALP media, pH 7.4, at 38.5 °C for 1 hour under 5% CO2 in the absence and presence of L-penicillamine, n-acetyl cysteine and α-tocopherol acetate. The sperm kinematics was studied using computer-assisted semen analyzer (CASA). The results revealed that 0.25 mM Lpenicillamine could increase the total motility, progressive motility, rapid motility, amplitude of lateral head displacement (ALH) and beat cross frequency (BCF) of buffalo sperm as compared to the control group. Similarly, 600 µM -tocopherol acetate could increase the total motility, rapid motility, curvilinear velocity, straight-line velocity, ALH and BCF of sperm. However, n-acetyl cysteine at the tested concentrations (0.125-1.0 mM) could not increase the sperm kinematics parameters. Thus, Lpenicillamine and α-tocopherol acetate were found to be two promising additives for improving buffalo sperm motility. However, effect of pre-freeze addition of these additives to the semen extender on postthaw motility of buffalo sperm warrants further investigation.ICAR-NIAN

Effect of urea supplemented and urea treated straw based diet on milk urea concentration in crossbred Karan-Fries cows

The study was undertaken to evaluate the effect of urea supplemented and urea treated straw based diet on milk ureaconcentration. Six multiparous crossbred Karan-Fries (Holstein Friesian ✕ Tharparkar) cows were blocked into threegroups of nearly equal body weight, DIM, milk yield and milk fat content and were randomized into a 3 ✕ 3 Latin squaredesign with 3-week period. Three experimental diets were fed to the animals. Composition of these diets were: Diet 1)green maize, wheat straw and concentrate mixture; Diet 2) green maize, wheat straw, concentrate mixture (urea supplemented)and molasses; Diet 3) green maize (4 % of total DM), 4 % urea treated wheat straw and concentrate mixture.Intake of DM and CP did not vary across the diets. Intake of digestible crude protein (DCP) was found significantlyhigher in diet 2, while ME and NEL intakes were found significantly lower in diet 3 but did not differ between diets 1and 2. Average milk and plasma urea concentrations (mg dl-1) were found 29.2 ± 2.6, 45.3 ± 0.9, 34.5 ± 2.3 and 28.9± 2.4, 36.6 ± 1.4, 33.9 ± 2.2, respectively in diet 1, diet 2 and diet 3. Urea concentrations in morning milk sampleswere found significantly lower than noon or evening samples in all the three diets. Concentrations of urea in milk andplasma were found closely correlated (r = 0.94) and the regression equation developed was, plasma urea = 8.90 (.89)+ .79 (.02) milk urea. Intake (g) of DCP than CP, per unit (MCal) of ME was found more closely associated with milk ureaconcentration. The study revealed that urea supplementation and urea treated straw based diet increased urea concentrationsignificantly in milk and plasma. Morning milk urea values that estimated at a time gap of 15 hr since last majorfeeding may be considered as the lowest level and can be used for interpretation to monitor feeding adequacy or reproductiveperformances in dairy cows

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Not AvailableA study was conducted in May 2003 to characterize plasma growth hormone (GH) pattern in growing mithuns (Bos frontalis), a rare semi-wild ruminant. Six mithun calves averaging 235 day of age and 124 kg were maintained in semi-intensive system and group-fed once daily. Animals gained at a mean rate of 0.54 kg/day, with individuals ranging from 0.34 to 0.66 kg/day. Blood samples collected at 15-minute intervals starting from 0600h for nine-hour period were assayed for plasma GH. Growth hormone patterns consisted of frequent pulses of varying amplitude. Growth hormone pulses occurred at an average frequency of 0.69/h, the rate did not differ markedly among mithuns nor hour of day. The magnitude of GH secretory pulses varied significantly among mithuns. Growth hormone peaks averaged 95.0 and 45.2 ng/ml in mithuns having the highest and lowest GH peaks, respectively. Peak and mean GH levels were associated positively (r=0.98, P<0.001) and both were associated negatively (r=–0.97 and –0.98, respectively; P<0.01) with rates of gain. Results from the study show that (1) GH peaks occur at frequent intervals throughout the sampling period and (2) alteration in GH levels and patterns are elicited more by pulse amplitude than frequency modulation.Not Availabl

ICAR- NIANP News letter, Jul-Dec, 2014

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Methane mitigation potential of phyto-sources from Northeast India and their effect on rumen fermentation characteristics and protozoa in vitro

Aim: The aim of the study was to explore the anti-methanogenic potential of phyto-sources from Northeast region of the country and assess the effect on rumen fermentation characteristics and protozoa for their likely inclusion in animal diet to reduce methane emission. Materials and Methods: Twenty phyto-sources were collected from Northeast state, Assam, during March to April 2014. Phyto-sources were analyzed for their tannin content followed by screening for methane mitigation potential using in vitro system. The effect of tannin on methane production and other fermentation parameters was confirmed by attenuating the effect of tannin with polyethylene glycol (PEG)-6000 addition. About 200 mg dried phyto-source samples were incubated for 24 h in vitro, and volume of gas produced was recorded. The gas sample was analyzed on gas chromatograph for the proportion of methane in the sample. The effect of phyto-sources on rumen fermentation characteristics and protozoal population was determined using standard methodologies. Results: Results from studies demonstrated that Litchi chinensis, Melastoma malabathricum, Lagerstroemia speciosa, Terminalia chebula, and Syzygium cumini produced comparatively less methane, while Christella parasitica, Leucas linifolia, Citrus grandis, and Aquilaria malaccensis produced relatively more methane during in vitro incubation. An increase (p<0.05) in gas and methane production from the phyto-sources was observed when incubated with PEG-6000. Entodinimorphs were prominent ciliates irrespective of the phyto-sources, while holotrichs represented only small fraction of protozoa. An increase (p<0.05) in total protozoa, entodinimorphs, and holotrichs was noted when PEG-6000 added to the basal substrate. Our study confirmed variable impact of phyto-sources on total volatile fatty acid production and ammonia-N. Conclusion: It may be concluded that L. chinensis, M. malabathricum, L. speciosa, S. cumini, and T. chebula are having potent methane suppressing properties as observed in vitro in 24 h. These leaves could be supplemented in the animal diet for reducing methane emission; however, in vivo trials are warranted to confirm the methane inhibitory action and optimize the level of supplementation

ICAR- NIANP News letter, Jan-Jun, 2016

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