A recombinant-based feline immunodeficiency virus antibody enzyme-linked immunosorbent assay.

We have developed an antibody detection enzyme-linked immunosorbent assay (ELISA) for the identification of animals infected by feline immunodeficiency virus (FIV). The ELISA solid-phase antigen consists of recombinant FIV gag proteins expressed in bacteria. The proteins are purified from bacterial lysates as insoluble inclusion bodies. In the case of bacterially expressed p24gag, it is shown that all of the linear, sequential epitopes presented by viral p24 during infection are retained. Purified preparations can be substituted for solid-phase whole virus in the IDEXX PetChektm immunoassay. The antibody ELISA duplicates the sensitivity and specificity of the whole virus based PetChek plate assay.     

玉米F_2群体中偏分离RFLP标记在染色体上的分布及原因分析

研究了利用玉米85个 DNA 限制性片段长度多态性(RFLP)标记在玉米自交系7922与5003杂交 F2群体中的分离。结果表明,8.33%的标记显著或极显著地偏离了预期的孟德尔比例而表现偏分离,发现了2个偏分离的染色体热点,即第1染色体上的 umc128-umc107和第3染色体上的 asg24-asg48,其偏分离形成的主要原因是存在配子体选择。     

Uterine immune reaction and reproductive performance of sows inseminated with extended semen and infused with pooled whole dead semen

The objective of this study was to investigate the effect of infusing whole dead semen (WDS) after AI with diluted commercial semen on uterine inflammatory reaction and embryonic survival rate in gilts. Sixty Yorkshire-Landrace gilts were assigned at their second estrus to one of the following AI treatments: 1) commercial semen adjusted to 1 x 10(9) sperm cells (S1) per dose, followed by an infusion of 80 mL of WDS (S1-WDS); 2) S1 followed by an infusion of 80 mL of Beltsville Thawing Solution (S1-BTS); 3) commercial semen adjusted to 3 x 10(9) sperm cells (S3) per dose, followed by an infusion of 80 mL of BTS (S3-BTS); and 4) a negative control group, in which gilts received two infusions of 80 mL of BTS (BTS). Two days after the first AI, eight gilts from Groups 1, 2, and 4 were slaughtered and reproductive tracts were collected. One horn was cut open longitudinally along the antimesometrial aspect and endometrial samples were taken and immediately frozen for analysis of messenger RNA (mRNA) abundance for inflammatory cytokines and growth factors. The other horn was flushed with 20 mL of PBS, and the contents of interferon-gamma (IFN-gamma), tumor necrosis factor-alpha (TNF-alpha) and transforming growth factor-beta1 (TGF-beta1) were determined by ELISA. On d 25 after AI, gilts from Groups 1, 2, and 3 were slaughtered and their reproductive tracts were collected to evaluate the number of fetuses and corpora lutea. On d 2 after the first AI, only TGF-beta1 was detected in the flush of all gilts, and no difference was observed between S1-WDS, S1-BTS, and BTS gilts. Endometrial levels of IFN-gamma and interleukin (IL)-6 mRNA were marked in all gilts, but they were not affected by the AI treatments, whereas the mRNA abundances for IL-1 and IL-2 were negligible. Infusions of WDS or BTS after a fertile AI did not affect IGF-I, IGF-I receptor, or IGF-II mRNA levels compared with gilts infused with BTS only, whereas the mRNA abundance for the IGF-II receptor was decreased (P < 0.05) in WDS-infused gilts. In gilts inseminated with S1 doses, infusion of WDS did not affect the number of live embryos. Although infusions of WDS did not affect the mRNA level and secretion of the cytokines measured and did not improve embryonic survival rates, further studies are needed to better understand the influence of semen composition on the uterine response after mating.     

Long term impact of mineral amendment on the distribution of the mineral weathering associated bacterial communities from the beech Scleroderma citrinum ectomycorrhizosphere

Liming is a known forest management procedure used to amend nutrient-poor soils such as soils of acidic forests to rectify cation deficiencies and to restore soil pH. However, although this procedure is well known for its beneficial effect on the forest trees, its relative impact on the functional and taxonomic diversity of the soil bacterial communities has been poorly investigated. In this study, we characterized the ability of the soil bacteria to weather soil minerals and to hydrolyze chitin. A collection of 80 bacterial strains was isolated from the Scleroderma citrinum ectomycorrhizosphere and the adjacent bulk soil in two stands of mature beeches (Fagus sylvatica) developed on very acidic soil and presenting two levels of calcium (Ca) availability: a control plot as well as a plot amended with Ca in 1973. All the bacterial isolates were identified by partial 16S rRNA gene sequence analysis as members of the genera Burkholderia, Bacillus, Dyella, Kitasatospora, Micrococcus, Paenibacillus, Pseudomonas, and Rhodanobacter. Using a microplate assay for quantifying the production of protons and the quantity of iron released from biotite, we demonstrated that the bacterial strains from the amended plot harbored a significant higher mineral weathering potential that the ones isolated from the control plot. Notably, the weathering efficacy of the ectomycorrhizosphere bacterial isolates was significantly greater than that of the bulk soil isolates in the control treatment but not in the amended plot. These data reveal that forest management, here mineral amendment, can strongly affect the structure of bacterial communities even over the long term.     

Impact of diet deprivation and subsequent over-allowance during prepuberty. Part 1. Effects on growth performance, metabolite status, and mammary gland development in gilts

The impact of diet deprivation and subsequent over-allowance on the metabolite status, mammary development, and mammary gene expression in prepubertal gilts was determined. Forty-seven gilts were reared under a conventional (control, CTL; n = 23) or an experimental (treatment, TRT; n = 24) dietary regimen. The later regimen (consisting of diet deprivation and subsequent over-allowance) provided 70 (restriction diet, RES) and 115% (over-allowance diet, OVER) of the protein and DE contents provided by the CTL diet. Experimental diets were fed ad libitum starting at 27.7 ± 3.4 kg of BW as follows: 3 wk RES, 3 wk OVER, 4 wk RES, and 4 wk OVER. At each diet change, BW and individual feed intakes were measured, and blood samples for metabolite and IGF-I assays were obtained. Some gilts (11 CTL and 12 TRT) were slaughtered on d 235 (after reaching puberty) to collect mammary tissue for compositional analyses and measures of gene expression. Body weight gain (P < 0.01) and G:F (P < 0.05) of gilts were reduced during each period with the RES diet; however, there was no compensatory growth during the periods when the OVER diet was fed. Feeding the RES diet reduced concentrations of urea and IGF-I (P < 0.01) and feeding the OVER diet increased FFA (P < 0.01) and glucose (P < 0.10) in TRT gilts compared with CTL gilts. The TRT gilts had less parenchymal tissue (P < 0.05) and tended to have less total parenchymal fat and protein (P < 0.10) than CTL gilts. The mammary mRNA relative abundance of the signal transducers and activators of transduction 5B was decreased in TRT compared with CTL gilts (P < 0.05). In conclusion, the diet deprivation and over-allowance regimen used in the growing-finishing period did not have beneficial effects on mammary development after puberty. In fact, a detrimental effect was observed.     

Adiponectin enhances in vitro development of swine embryos

Recent studies have shown that factors from adipose tissue influence and regulate the reproductive system. Hormones such as leptin and resistin are now known to regulate several reproductive processes. Adiponectin is the most abundant protein secreted by adipose tissue, and its circulating concentration is inversely related to adiposity and body mass index. Little is known about the involvement of adiponectin in reproduction. In the present study, the effect of recombinant adiponectin on the meiotic maturation and early embryo development in vitro was investigated, using porcine oocytes. Adiponectin receptors, AdipoR1 and AdipoR2, were found to be expressed in porcine oocytes and cumulus cells of both small and large follicles. Both AdipoR1 and AdipoR2 were immunolocalized to cumulus-oocyte complexes (COCs), oocytes, and early developing embryos. When included in oocyte maturation medium for 46 h, adiponectin significantly decreased the frequency of meiotic immature oocytes derived from large follicles (3-6 mm) but not from small follicles (<3mm). From studies of oocytes matured in the presence of adiponectin and mitogen-activated protein kinase (MAPK) pathway inhibitors MEK1 (PD98059), MEK1/2 (U0126), and p38MAPK (SB203580) it was concluded that adiponectin enhances oocyte maturation thought the p38MAPK pathway. Finally, a superior rate of embryo development to the blastocyst stage was achieved by embryos cultured in the presence of adiponectin. These results indicate that adiponectin has a positive effect on the meiotic maturation and in vitro embryo development of porcine oocytes and suggests a physiological role for this adipokine in early development in mammals.     

Mammary gland development and hormone levels in pregnant Upton-Meishan and large white gilts

Genetic differences between Upton-Meishan (UM, n = 13) and Large White (LW, n = 14) gilts were studied with regard to mammary gland development and concentrations of hormones. Gilts were weighed and their backfat measured at mating, and at d 70 and 109 of gestation. Jugular blood samples were also collected at these times and assayed for prolactin, cortisol, IGF-I, insulin, glucose, progesterone, and estradiol. Gilts were slaughtered on d 110 of gestation. One row of mammary glands was used for dissection and biochemical analyses. The other row was used for determination of prolactin receptor number and affinity. UM gilts weighed less (P<0.05) and had more backfat (P<0.01) than LW gilts at all times. Parenchymal tissue weight was less (P<0.05) in UM gilts. Percent fat (P<0.001) and dry matter (P<0.001) in parenchymal tissue were greater in UM gilts while that of protein (P<0.001) was lower. Total protein weight in parenchyma was also lower in parenchyma was also lower in UM gilts (P = 0.01). Both DNA (P<0.001) and RNA (P<0.001) contents were lower in UM gilts while RNA/DNA remained similar (P>0.1). Number of prolactin receptors were lower (P = 0.06) and affinity greater (P<0.05) in UM gilts. Cortisol levels were greater (P<0.01) in UM gilts while other hormones were not affected (P> 0.1). Results clearly demonstrate genetic differences with regard to mammogenesis in gilts and suggest that the less mammary gland development in Upton-Meishan compared with Large White breed of gilts may be related to lower number of prolactin receptors.