Molecular characterization and expression profile of interferon-stimulated gene 15 (ISG15) in the endometrium of goat (Capra hircus).

Interferon-stimulated gene 15 (ISG15), a ubiquitin-like protein, is responsible for uterine receptivity, implantation and conceptus development in different ruminant species, but in goat (Capra hircus) its role is yet to be explicated. In the present study, the ISG15 gene was cloned, characterized and its temporal expression profile was examined in the endometrium of caprine (cp). A fragment of cpISG15 gene, 1033 bp in length, was amplified, cloned and sequenced from genomic DNA covering the coding region. Sequence analysis of cpISG15 gene revealed that it was comprised of two exons of 59 bp and 496 bp encoding a peptide of 157 amino acids. Complementary DNA (cDNA) and deduced amino acid sequences of cpISG15 exhibited 99 and 98, 93 and 88, 94 and 89, 76 and 66, and 73 and 62% identity with that of sheep, cattle, buffalo, human and mice, respectively. Further, relative expression of cpISG15 mRNA and protein was determined by quantitative real-time PCR (qPCR) and Western blot, respectively, in the endometrium of pregnant and cyclic does. Both cpISG15 mRNA and protein were expressed maximally (P < 0.05) in the endometrium during early stage of pregnancy (16-24 d) as compared to cyclic does, but no significant difference was observed in cpISG15 mRNA and protein expression in the endometrium between the later stage of pregnancy (25-40 d) and cyclic does. It is concluded that cpISG15 is almost similar in structure and probably in function also to other species as it has been found significantly upregulated during early pregnancy.

Short-term feed and light deprivation reduces voluntary activity but improves swimming performance in rainbow trout Oncorhynchus mykiss.

Rainbow trout Oncorhynchus mykiss (~ 180 g, 16 °C and < 5 kg m-3) that were feed deprived and kept in total darkness showed a significant increase in critical swimming speed (U crit) between 1 and 12 days of deprivation (from 3.35 to 4.46 body length (BL) s-1) with no increase in maximum metabolic rate (MMR). They also showed a significant decrease in the estimated metabolic rate at 0 BL s-1 over 12 days which leads to a higher factorial aerobic metabolic scope at day 12 (9.38) compared to day 1 (6.54). Routine metabolic rates were also measured in ~ 90 g rainbow trout that were swimming freely in large circular respirometers at 16 °C. These showed decreasing consumption oxygen rates and reductions in the amount of oxygen consumed above standard metabolic rate (a proxy for spontaneous activity) over 12 days, though this happened significantly faster when they were kept in total darkness when compared to a 12:12-h light-dark (LD) photoperiod. Weight loss during this period was also significantly reduced in total darkness (3.33% compared to 4.98% total body weight over 12 days). Immunological assays did not reveal any consistent up- or downregulation of antipathogenic and antioxidant enzymes in the serum or skin mucus of rainbow trout between 1 and 12 days of feed and light deprivation. Overall, short periods of deprivation do not appear to significantly affect the performance of rainbow trout which appear to employ a behavioural energy-sparing strategy, albeit more so in darkness than under a 12:12-h LD regime.

Thermal plasticity of skeletal muscle mitochondrial activity and whole animal respiration in a common intertidal triplefin fish, Forsterygion lapillum (Family: Tripterygiidae).

Oxygen demand generally increases in ectotherms as temperature rises in order to sustain oxidative phosphorylation by mitochondria. The thermal plasticity of ectotherm metabolism, such as that of fishes, dictates a species survival and is of importance to understand within an era of warming climates. Within this study the whole animal O2 consumption rate of a common New Zealand intertidal triplefin fish, Forsterygion lapillum, was investigated at different acclimation temperatures (15, 18, 21, 24 or 25 °C) as a commonly used indicator of metabolic performance. In addition, the mitochondria within permeabilised skeletal muscle fibres of fish acclimated to a moderate temperature (18 °C Cool acclimation group-CA) and a warm temperature (24 °C. Warm acclimation group-WA) were also tested at 18, 24 and 25 °C in different states of coupling and with different substrates. These two levels of analysis were carried out to test whether any peak in whole animal metabolism reflected the respiratory performance of mitochondria from skeletal muscle representing the bulk of metabolic tissue. While standard metabolic rate (SMR- an indicator of total maintenance metabolism) and maximal metabolic rate ([Formula: see text]O2 max) both generally increased with temperature, aerobic metabolic scope (AMS) was maximal at 24 °C, giving the impression that whole animal (metabolic) performance was optimised at a surprisingly high temperature. Mitochondrial oxygen flux also increased with increasing assay temperature but WA fish showed a lowered response to temperature in high flux states, such as those of oxidative phosphorylation and in chemically uncoupled states of respiration. The thermal stability of mitochondria from WA fish was also noticeably greater than CA fish at 25 °C. However, the predicted contribution of respirational flux to ATP synthesis remained the same in both groups and WA fish showed higher anaerobic activity as a result of high muscle lactate loads in both rested and exhausted states. CA fish had a comparably lower level of resting lactate and took 30 % longer to fatigue than WA fish. Despite some apparent acclimation capacity of skeletal muscle mitochondria, the ATP synthesis capacity of this species is constrained at high temperatures, and that a greater fraction of metabolism in skeletal muscle appears to be supported anaerobically at higher temperatures. The AMS peak at 24 °C does not therefore represent utilisation efficiency of oxygen but, rather, the temperature where scope for oxygen flow is greatest.  

Hormone profile of crossbred goats during the periparturient period.

The plasma hormone profile of progesterone, 17 beta-oestradiol, prolactin, growth hormone (GH), cortisol, triiodothyronine (T3), thyroxine (T4) and insulin were estimated by radioimmunoassay in jugular blood samples from 10 Alpine x Beetal crossbred goats during the periparturient period. The plasma progesterone concentrations declined from day 20 prior to kidding to the day of kidding and then remained at a basal level. 17 beta-Oestradiol concentrations declined from day 20 to day 4 prepartum, followed by a continuous increase to reach a peak at kidding. Plasma oestradiol declined abruptly on day 1 postpartum and remained low throughout the postpartum period. The plasma prolactin concentrations increased continuously from day 20 to day 10 prepartum but then increased to a peak on the day of kidding, followed by a transient decline by day 5 postpartum. The plasma GH concentrations varied significantly (p < 0.01) on the different days of sampling but were highest on the day of kidding. The plasma cortisol concentration declined from day 20 to day 2 prepartum, after which it abruptly increased on day 1 prepartum to reach a peak on the day of kidding. The concentrations of plasma T3 remained similar during the prepartum period but were minimal on the day of kidding and increased during the postpartum period. The concentration of plasma T4 was also lowest on the day of kidding and increased during the postpartum period. Plasma insulin concentrations were low in the day of kidding.

Changes in blood glucose, plasma non-esterified fatty acids and insulin in pregnant and non-pregnant goats.

The blood glucose and the plasma non-esterified fatty acids (NEFA) and insulin concentrations were estimated in jugular blood samples from 18 Alpine x Beetal and Sannen x Beetal goats during pregnancy and compared with samples from non-pregnant goats and from goats during the periparturient period. The blood glucose levels in the pregnant goats rose to a peak of about 60 +/- 1.36 mg/ml at 42-56 days and then declined to about 46 +/- 2.37 mg/ml at 112-126 days. In non-pregnant goats, the blood glucose levels were significantly (p < 0.01) higher than in pregnant goats, except between days 42 and 70 (59 +/- 1.36 mg/ml). On the day of kidding, the levels declined significantly (p < 0.01), increasing again thereafter. The plasma NEFA concentrations were significantly higher in pregnant than in non-pregnant goats from days 56 to 126. The NEFA concentration increased on the day of kidding, followed by a transient fall by day 3. The plasma insulin concentration was usually higher in pregnant than in non-pregnant goats, except between days 56 and 70 and from day 126 onwards. The insulin concentration fell late in pregnancy, but there was a transient increase 2 days after parturition. The blood glucose and plasma NEFA concentrations can be used as indices of nutritional status during pregnancy in goats.

DNA amplification assay for rapid detection of bovine tubercle bacilli in semen.

Tubercle bacilli shed in the semen can be a potential hazard for unlimited number of cows through artificial insemination. We have evaluated the efficacy of a DNA amplification technique by polymerase chain reaction (PCR) for the detection of tubercle bacilli in fresh and frozen semen using spiked samples. The test was based on insertion sequence IS 1081 and could detect as low as 10-100 bacterial cells per ml of spiked semen. The specificity of the test was 100%. The method was applied to semen samples from known and suspected tuberculous bulls. Each of 20 semen samples (fresh and frozen diluted) from one of the three breeding bulls included in the study was found positive while the remaining 40 samples from the other two bulls failed to generate any detectable signal. PCR products were confirmed with Southern blot hybridization to an alpha 32P labeled-PCR product of the target sequence from the IS 1081 element of the Mycobacterium tuberculosis complex.