Production of Lambs After the Transfer of Fresh and Cryopreserved in vitro Produced Embryos from Prepubertal Lamb Oocytes and Unsorted and Sex-sorted Frozen-thawed Spermatozoa

Cumulus-oocyte complexes from hormone-stimulated 3-4-week-old (n=43) and 6-7-week-old (n=12) prepubertal lambs were matured in vitro and incubated with unsorted, or X- or Y-spermatozoa separated with a high-speed cell sorter (SX MoFlo)frozen-thawed. Presumptive zygotes were then cultured to the blastocyst stage, and transferred to recipients fresh or after cryopreservation (frozen). Oocyte cleavage was higher (p <0.05) with unsorted (515/926, 55.6%) than X- or Y-spermatozoa (261/672, 38.8% and 229/651, 35.2%, respectively) and blastocyst formation (% zygotes) by Day 9 of in vitro culture was lower (p <0.05) for X- (102/261, 39.1%) than unsorted spermatozoa (249/515, 48.3%), but did not differ between Y-spermatozoa (103/229, 45.0%) and unsorted spermatozoa, or between X- and Y-spermatozoa (p >0.05). For fresh embryos, survival to term was 50.0% (3/6) for unsorted, 0.0% (0/6) for X- and 16.7% (1/6) for Y-spermatozoa-derived embryos (p >0.05), and for frozen embryos was 4.0% (2/50) for unsorted, 9.1% (2/22) for X- and 2.9% (1/34) Y-spermatozoa-derived embryos (p >0.05). Of the two lambs born from X-spermatozoa-derived embryos, one was female (50%), and from the two Y-spermatozoa-derived lambs, both were male (100%), demonstrating that lambs can be produced after the transfer of fresh and cryopreserved IVP embryos derived from prepubertal lamb oocytes and frozen-thawed sex-sorted sperm.     

Treatment of anovulatory anoestrous postpartum dairy cows with a gonadotropin-releasing hormone (GnRH), prostaglandin F2 alpha, GnRH regimen or with progesterone and oestradiol benzoate

AIM: To compare 2 treatments for anovulatory anoestrus (AA) in postpartum dairy cows. The treatments were combinations of gonadotropin-releasing hormone (GnRH) and prostaglandin F2 (PG) or progesterone (P4) and oestradiol benzoate (ODB). METHODS: Forty AA cows from each of 5 herds were blocked by age (2 or 2 years old) and randomly assigned to 1 of 2 treatments. The first group (GPG) were treated with 250 mug of a GnRH analogue, gonadorelin, followed 7 days later by 15 mg of the PG analogue, luprostiol. Two days later the cows were injected with 250 mug of gonadorelin. Cows were artificially inseminated 16-24 h after the second GnRH injection. The second group (P4+ODB) were treated with an intravaginal P4 releasing device for 6 days, followed 24 h after device removal by injection of 1 mg of ODB. Cows were pregnancy tested 35-40 days after the initial insemination and twice again at 6-8 week intervals thereafter. RESULTS: There was no significant difference between P4+ODB and GPG groups in the percentage of cows submitted for insemination in the first 7 days (94.0% vs 100% for P4+ODB vs GPG, respectively; p>0.3), in conception rate to first insemination within the first 7 days (43.6% vs 35.0% for P4+ODB vs GPG, respectively; p>0.2), in the percentage of cows conceiving in the first 28 days of the breeding period (68.0% vs 58.3%, P4+ODB vs GPG, respectively; p>0.1), or in median interval from the end of treatment to conception (20 vs 21 days; p>0.1). CONCLUSIONS: No differences in the reproductive performance of AA cows treated with either P4+ODB or GPG were detected. However, given the small number of animals enrolled, further data are required before the GPG protocol can be recommended for treatment of AA cows.     

Device Applications of Side-Chain Ferroelectric Liquid Crystalline Polymer Films

Side-chain ferroelectric liquid crystalline polymers are currently used in a number of applications, including displays and electrical sensors. Comparisons with existing technologies and materials indicate that relative to ceramics, such polymers have lower figures of merit but offer greater durability in sensor applications.     

Impact of Distillery Effluent on Soil Carbon and Nitrogen Dynamics in a Vertisol of Central India

Distilleries produce a huge quantity of effluents, popularly known as spent wash (SW), which when bio-methanated produce post-methanation effluents (PME). A field experiment on soybean–wheat system was conducted for five consecutive years in a Vertisol of central India to evaluate the effect of distillery effluent (DE) on soil carbon and nitrogen dynamics. Ten treatment combinations consisting of control, 100% NPK + Farmyard Manure (FYM), and graded level of SW and PME were applied. Total carbon content of soil increased significantly with applications of FYM and DE. SW was found superior in enhancing carbon content of soil in comparison to PME. Farmyard Manure contributed more carbon toward the recalcitrant pool, whereas DE contributed more carbon toward the active and slow pool. Nitrogen (N) availability was significantly improved with the application of DE. Balanced application of DE may act as amendment for increasing C and N stocks in Vertisol.     

Immunological characterization of the equine airway epithelium and of a primary equine airway epithelial cell culture model

Our understanding of innate immunity within the equine respiratory tract is limited despite growing evidence for its key role in both the immediate defense and the shaping of downstream adaptive immune responses to respiratory disease. As the first interface to undergo pathogen invasion, the respiratory epithelium is a key player in these early events and our goal was to examine the innate immune characteristics of equine respiratory epithelia and compare them to an in vitro equine respiratory epithelial cell model cultured at the air-fluid interface (AFI). Respiratory epithelial tissues, isolated epithelial cells, and four-week old cultured differentiated airway epithelial cells collected from five locations of the equine respiratory tract were examined for the expression of toll-like receptors (TLRs) and host defense peptides (HDPs) using conventional polymerase chain reaction (PCR). Cultured, differentiated, respiratory epithelial cells and freshly isolated respiratory epithelial cells were also examined for the expression of TLR3, TLR9 and major histocompatibility complex (MHC) class I and class II using fluorescence-activated cell sorting (FACS) analysis. In addition, cytokine and chemokine profiles from respiratory epithelial tissues, freshly isolated respiratory epithelial cells, and cultured, differentiated, epithelial cells from the upper respiratory tract were examined using real-time PCR. We found that respiratory epithelial tissues and isolated epithelial cells expressed TLRs 1-4 and 6-10 as well as HDPs, MxA, 2'5' OAS, β-defensin-1, and lactoferrin. In contrast, epithelial cells cultured at the AFI expressed TLRs 1-4 and 6 and 7 as well as MxA, 2'5' OAS, β-defensin-1, but had lost expression of TLRs 8-10 and lactoferrin. In addition, MHC-I and MHC-II surface expression decreased in epithelial cells cultured at the AFI compared to isolated epithelial cells whereas TLR3 and TLR9 were expressed at similar levels. Lastly, we found that equine respiratory epithelial cells express an array of pro-inflammatory, antiviral and regulatory cytokines and that after four weeks of in vitro growth conditions, equine respiratory epithelial cells cultured at the AFI retained expression of GM-CSF, IL-10, IL-8, TGF-β, TNF-α, and IL-6. In summary, we describe the development of an in vitro equine respiratory epithelial cell culture model that is morphologically similar to the equine airway epithelium and retains several key immunological properties. In the future this model will be a used to study equine respiratory viral pathogenesis and cell-to-cell interactions.     

Fate of (14)C-labeled carbonyl sulfide on grains and grain fractions

14C-Labeled carbonyl sulfide (COS) was used to measure the amount of sorbed fumigant and alteration products on grains. Wheat, paddy rice, polished rice, mungbean, and safflower were exposed to a 60 mg L(-)(1) of (14)COS for 7 days and then aired for 5 days. Carbonyl sulfide and/or alteration products in sugars, protein, starch, amino acids, protopectines, and hemicelluloses were undetectable. The total uptakes of radioactivity determined after fractionation and also by extraction were in the range of 36-53 ng g(-)(1) COS equiv. The total radioactivity determined by autoradiography was below the detection limit of 70 ng g(-)(1) COS equiv. Radioactivity in the commodities was less than 0.003% of all the radioactivity applied. The amount of retained radiolabel was measured in three ways. First, biochemical fractions such as lipids and amino acids were separated by chromatography, and the activity was determined in each component. Second, commodities were crushed and extracted in aqueous acetone until the maximum amount of radiolabel was extracted. Third, autoradiography was carried out on commodity kernels.