Differences in Reproductive Performance, Embryo Development, Interferon-tau Secretion by the Conceptus and Luteal Function in Ewe Lambs Synchronized in Oestrus before or after the Spontaneous Onset of Luteal Activity Preceding Puberty

In mid-September, 1 month before the insertion of intravaginal pessaries to induce sexual activity, blood samples were collected every 4 days from 16 ewe lambs aged 7 months, in order to determine the incidence of ovulations by measurement of plasma progesterone concentrations. It has been studied whether the response to a progestagen treatment of ewe lambs apparently close to puberty could be modified by the onset of the ovarian events preceding puberty. The effect of the presence or absence of ovulations prior to progestagen treatment on the potential reproductive performance (fertility, litter size and fecundity), embryo development [embryo quality and interferon-tau (IFNτ) secretion], luteal function (progesterone secretion in vitro ) and endometrial progesterone content was studied in seven ovulating (Ov+) and nine nonovulating ewe lambs (Ov−) on day 14 after mating. The best potential reproductive results were obtained with Ov+ animals, although these differences could not be initially attributed to either different progesterone secretion in vitro or concentration of endometrial progesterone. Irrespective of the experimental groups, secretion of progesterone by luteal tissue from ewe lambs with normal embryos was significantly greater (p<0.05) than that of animals with abnormal embryos or with no embryos. Normal embryos secreted a higher amount of IFN-τ than those embryos classified as abnormal (p<0.07). In conclusion, ewe lambs which exhibit luteal activity before puberty have the highest levels of reproductive performances after a progestagen treatment. Corpora lutea from ewe lambs with normal embryos had higher rates of progesterone secretion in vitro and their embryos had a higher IFN-τ production by the embryos, indicating greater capacity for subsequent development.     

Early pregnancy alters the metabolic responses to restricted nutrition in sheep

This study investigated whether a 27-day period of nutrition at half-maintenance during early pregnancy (up to Day 14) could alter maternal endocrine responses. Forty-six ewes were fed all or half of their maintenance requirements and slaughtered on Day 14 of the oestrous cycle or pregnancy. We used real time RT-PCR to study gene expression of growth hormone receptor (GHR) and leptin in adipose tissue and GHR, GHR1A and of the insulin-like growth factor I (IGF-I) in the liver. Blood profiles of metabolites and metabolic hormones were also determined. Throughout the experiment, underfed animals presented lower body weight and body condition, greater plasma concentrations of non-esterified fatty acids (NEFA), and lower plasma concentrations of leptin, compared to adequately fed animals. Undernutrition affected the patterns of gene expression in adipose and hepatic tissues, and the responses differed between pregnant and non-pregnant ewes. In adequately fed ewes, pregnancy up-regulated leptin mRNA expression in adipose tissue, a response that was impaired in underfed ewes. The hepatic expression of IGF-I mRNA was increased by pregnancy in underfed animals while no effect was observed in adequately fed ewes. It remains to be determined whether the changes in the endocrine milieu are paralleled by modifications in uterine gene expression that could alter the environment of the embryo during early pregnancy.     

The effect of being housed with adult female flock-mates on the age at puberty of autumn-born ewe-lambs

Veterinary Research Communications - Twenty-five ewe-lambs born in November were used to measure the effect of the presence of cyclic or non-cyclic adult ewes on the age at first fertile estrus....     

Alternative methodologies to estimate ingestion of caecotrophes in growing rabbits

This study was carried out in order to estimate caecotrophe intake in growing rabbits by three existing procedures: caecotrophes collection after collar fitting, urinary purine derivatives (PD) excretion and microbial ¹⁵N-lysine incorporation. In a first experiment sixteen New Zealand White male rabbits were divided in three groups receiving the same diet, but supplemented with ¹⁵NH₄Cl in the first group (T₁: 6 rabbits). The second group (T₂: 6 rabbits) was also fed the labelled diet but only during the last ten days of the fattening period when animals were fitted a neck collar to prevent caecotrophy. The third group (T₃: 4 animals) received the basal diet and was used as control. In two additional trials the daily contribution to urinary excretion of endogenous purine compounds (469 ± 50.8 μmol/W^0.75) and creatinine excretion (807 ± 127.6 μmol/W^0.75) were determined. The highest estimation of microbial nitrogen recycling was obtained by the urinary PD method (0.79 ± 0.096 g/d), whereas caecotrophes collection and ¹⁵N-lysine incorporation methods showed similar values (0.49 ± 0.049 and 0.45 ± 0.015 g/d, respectively). Our results seem to indicate an overestimation of microbial nitrogen recycling in growing rabbits by PD methodology, while neck collar fitting procedure gave similar results, although more variable than microbial ¹⁵N-lysine incorporation.     

Development of a Simple Enzyme Immunoassay for the Determination of Ovine Luteinizing Hormone

The present study describes the development and validation of a simple sensitive and specific sandwich enzyme immunoassay (EIA) for the quantification of ovine luteinizing hormone (LH) in plasma. Microtitre plates were coated with the capture antibody 518b7 anti-bovine LH. A second peroxidase-labelled anti-ovine LH antibody was used as tracer. A simple 3-step procedure was used for the sample analysis; (1) incubation of standards and samples with the pre-coated antibody plates for 2 h at 37^°C; (2) incubation with the peroxidase-labelled antibody for 1 h at room temperature; and (3) colour development with TMB substrate. A linear dose–response curve was obtained in the range 0–10 ng/ml (r ² > 0.99). The detection limit was 0.05 ng/ml, and the intra-assay and inter-assay coefficients of variation were 7% and 11.7%, respectively. The theoretical stability of microplates and reagents was calculated, this being greater than one year. Low or undetectable cross-reactivities were recorded for follicle-stimulating hormone, bovine thyroid-stimulating hormone, equine chorionic gonadotrophin and a gonadotrophin-releasing hormone (GnRH) analogue. The EIA was biologically validated by the determination of plasma LH concentrations of nine Rasa Aragonesa ovariectomized and estradiol-implanted ewes after a double GnRH challenge. In conclusion, this enzyme immunoassay provides an efficient, simple and sensitive method for the routine analysis of ovine LH.     

Effects of Melatonin Implants During Non‐Breeding Season on Sperm Motility and Reproductive Parameters in Rasa Aragonesa Rams

The effect of melatonin implants administered during non‐breeding season in Rasa Aragonesa rams on sperm motility parameters and other reproductive traits was assessed. In a first experiment, two Rasa Aragonesa rams were implanted (with melatonin group M), remaining other two males as control group (C). Semen of each group was collected from 1 May to 23 June, twice or three times a week, and motility parameters were assessed using a computer‐assisted sperm analysis system. Melatonin increased the percentage of progressive motile spermatozoa, particularly during 46–75 days after melatonin implantation (p < 0.01). In experiment 2, M and C in vitro fertilization ability had been determined by zona‐pellucida binding assays, using spermatozoa from experiment 1, obtained 60–70 days after melatonin was implanted. A significantly higher number of spermatozoa attached per oocyte was observed in frozen‐thawed immature ovine oocytes incubated with sperm from M animals than in those incubated with sperm from the C group (p < 0.01). Finally, a field assay (experiment 3) was performed. In this case, five Rasa Aragonesa rams were implanted with melatonin and three remained as control group. Sperm doses from those animals were used for artificial insemination of 2608 Rasa Aragonesa ewes from 39 different farms at non‐breeding season. Fertility, litter size and fecundity were studied. Semen from melatonin implanted rams seemed to increase both fertility and fecundity in ewes inseminated with spermatozoa obtained 46–60 days after implantation (p < 0.1). Thus, melatonin treatment in rams during non‐breeding season modifies sperm motility parameters and seems to improve the fertilization parameters obtained.     

抗菌添加剂纳米银粒子在断奶仔猪中的应用

试验旨在研究钠米银粒子在断奶猪饲粮中添加的可能性，共设计3个试验。试验1：将4头断奶7d仔猪的回肠内容物，分别与纳米银粒子0、25、50、100μg·g^-1在37℃下体外培养4h。结果显示，纳米银粒子（胶体形式）使大肠杆菌（P=0．003）和乳酸杆菌（P=0．041）浓度呈线性下降。但是，与对照组相比，乳酸杆菌比例无显著差异。试验2：将15头断奶猪分为3组，每组在饲粮中分别添加纳米银粒子0、20、40mg·kg^-1。结果表明，在断奶后的第2周，添加纳米银粒子组仔猪日增重呈线性增长（P=0．007）。通过培养计数法观测到回肠中大肠杆菌浓度呈线性减少趋势（P=0．073），但乳酸杆菌浓度没有受到影响。通过免疫荧光素原位杂交（FISH）法测得各处理组断奶仔猪回肠中大肠杆菌和乳酸杆菌浓度没有差异，但是，细菌总数（P＝0．010）和奇异菌（P=0．001）浓度逐渐减少。除了添加纳米银粒子20mg·^-1。组中梭状芽胞产气荚膜杆菌／梭菌胶原酶群浓度最低，其他细菌群均无显著差异。各处理组回肠黏膜组织学检测中也均无差异。试验3：为研究猪生产性能和其组织中银的残留量。将断奶后5周仔猪分成3个处理组，每个处理组6个重复，每个重复4头仔猪，每组饲粮分别添加纳米银粒子0、20、40mg·kg^-1。结果表明，添加纳米银粒子20mg·kg。组猪采食量最多（P〈0．05），但表观消化系数无差异。5周后，在仔猪骨骼肌和肾中没有银残留，但在肝脏中，纳米银粒子20mg·kg^-1组和40mg·kg^-1组中银残留量分别为1．354和2．445μg·kg。（肝脏干重）。可见，纳米银粒子作为一种饲料添加剂，可能通过其抗菌特性调节断奶仔猪采食量和生产性能。既可以抵制一定种类的菌群，也可以减少小肠微生物含量，且对于宿主新陈代谢的有利作用不可忽视。     

Study of the effect of presence or absence of protozoa on rumen fermentation and microbial protein contribution to the chyme

The aim of this study was to investigate the effect of presence or absence of protozoa on rumen fermentation and efficiency of microbial protein synthesis under different diets. Of 20 twin paired lambs, 1 lamb of each pair was isolated from the ewe within 24 h after birth and reared in a protozoa-free environment (n = 10), whereas their respective twin-siblings remained with the ewe (faunated, n = 10). When lambs reached 6 mo of age, 5 animals of each group were randomly allocated to 1 of 2 experimental diets consisting of either alfalfa hay as the sole diet, or 50:50 mixed with ground barley grain according to a 2 × 2 factorial arrangement of treatments. After 15 d of adaptation to the diet, the animals were euthanized and total rumen and abomasal contents were sampled to estimate rumen microbial synthesis using C(31) alkane as flow marker. Different ((15)N and purine bases) and a novel (recombinant DNA sequences) microbial markers, combined with several microbial reference extracts (rumen protozoa, liquid and solid associated bacteria) were evaluated. Absence of rumen protozoa modified the rumen fermentation pattern and decreased total tract OM and NDF digestibility in 2.0 and 5.1 percentage points, respectively. The effect of defaunation on microbial N flow was weak, however, and was dependent on the microbial marker and microbial reference extract considered. Faunated lambs fed with mixed diet showed the greatest rumen protozoal concentration and the least efficient microbial protein synthesis (29% less than the other treatments), whereas protozoa-free lambs fed with mixed diet presented the smallest ammonia concentration and 34% greater efficiency of N utilization than the other treatments. Although (15)N gave the most precise estimates of microbial synthesis, the use of recombinant DNA sequences represents an alternative that allows separate quantification of the bacteria and protozoa contributions. This marker showed that presence of protozoa decrease the bacterial-N flow through the abomasum by 33%, whereas the protozoa-N contribution to the microbial N flow increased from 1.9 to 14.1% when barley grain was added to the alfalfa hay. Absolute data related to intestinal flow must be treated with caution because the limitations of the sampling and maker system employed.