Abnormal water balance in a mutant strain of chickens

Polydipsia and polyuria are pronounced in chickens of a selected strain and this diabetes insipidus is inherited. The kidneys of such birds are capable of an antidiuretic response when lysine vasopressin or arginine vasotocin is injected. Osmotic pressure and sodium concentration of the plasmas of normal and mutant chickens are identical. Chicks predicted to have diabetes insipidus on the basis of parental pedigree are polydipsic.     

m‐carboxycinnamic acid bishydroxamide improves developmental competence,reduces apoptosis and alters epigenetic status and gene expression pattern in cloned buffalo (Bubalus bubalis) embryos

Incomplete or aberrant reprogramming of nuclear genome is one of the major problems in somatic cell nuclear transfer. In this study, we studied the effect of histone deacetylase inhibitor m‐carboxycinnamic acid bishydroxamide (CBHA) on in vitro development of buffalo embryos produced by Hand‐made cloning. Cloned embryos were treated with CBHA (0, 5, 10, 20 or 50 μM) for 10 hr from the start of reconstruction till activation. At 10 μM, but not at other concentrations examined, CBHA increased (p < .05) the blastocyst rate (63.77 ± 3.97% vs 48.63 ± 3.55%) and reduced (p < .05) the apoptotic index of the cloned blastocysts (8.91 ± 1.94 vs 4.36 ± 1.08) compared to untreated controls, to levels similar to those in IVF blastocysts (4.78 ± 0.74). CBHA treatment, at all the concentrations examined, increased (p < .05) the global level of H3K9ac in cloned blastocysts than in untreated controls to that observed in IVF blastocysts. Treatment with CBHA (10 μM) decreased (p < .05) the global level of H3K27me3 in cloned blastocysts than in untreated controls but it was still higher (p < .05) than in IVF blastocysts. CBHA (10 μM) treatment increased (p < .05) the relative expression level of pluripotency‐related genes OCT‐4 and NANOG, and anti‐apoptotic gene BCL‐XL, and decreased (p < .05) that of pro‐apoptotic gene BAX than in untreated controls but did not affect the relative expression level of apoptosis‐related genes p53 and CASPASE3 and epigenetics‐related genes DNMT1, DNMT3a and HDAC1. These results suggest that treatment of cloned embryos with 10 μM CBHA improves the blastocyst rate, reduces the level of apoptosis and alters the epigenetic status and gene expression pattern.     

Biology and status of timber rattlesnake (Crotalus horridus) populations in Pennsylvania

The purpose of this study was to examine certain aspects of the life history and ecology of the timber rattlesnake (Crotalus h. horridus) in Pennsylvania, and to evaluate the reported decline in numbers of this species. The influence of commercial hunting has been considered and suggestions are made for conservation of this unpopular but intriguing snake.The previous and present abundance of rattlesnakes was first considered by a programme of interviews with commercial hunters. Visits were also made to many supposed den sites in central Pennsylvania, and to four rattlesnake hunts held in the summer of 1976. There were consistent reports of a large recent decline in the number of rattlesnakes seen and captured. These opinions of the scarcity of rattlesnakes were personally confirmed by visits to known dens. One rattlesnake den was chosen for a study on dates of first capture in the spring, on transplantation of snakes from other localities, on movements of newly released and long-term den residents, and on rodent populations. During three consecutive springs the dates of first capture of rattlesnakes varied only between April 23 and 30, despite marked differences in the pattern of air and ground temperature variation. Many snakes transplanted to a new den site apparently left the area almost immediately. Six out of 33 adults released were recaptured at the new den; three of these hibernated there. Only one of the 48 newborn rattlesnakes released soon after birth in the autumn was recaptured in the subsequent spring. On emergence from hibernation two rattlesnakes were tracked by radiotelemetry. They remained at the den until June, and left soon after shedding. One snake was followed to a distance 1055 m from the den over a period of 69 days. Considerable rodent food was available on the talus slopes of this den. Mice (Peromyscuc leucopus) were abundant; in a 0·7 ha grid trapped for one week the population was estimated at 61/ha. Nine gravid rattlesnakes were held in the laboratory to obtain information on litter size. The average litter was seven (range 5–9) and females lost an average of 41% of their body weight during parturition. The metabolic demands of pregnancy are high and females only reproduce every two, or possibly every three, years. The smallest female to produce a litter had a snout-vent length of 770 mm, and it is estimated that this animal was five years old. Positive correlations between head length and snout-vent length, and body weight and snoutvent length were found. No significant differences in these relationships between the sexes or between snakes freshly caught in different areas were noted. There was a decline in relative body weight and head length after prolonged starvation in captivity. Male rattlesnakes have significantly longer tails than females. The timber rattlesnake in Pennsylvania is rapidly approaching extinction. Prohibition of bounties, snake hunts, and commercial sale of snakes, and protection of rattlesnakes in remote areas of state and national forests would significantly enhance the prospects for survival of viable populations of Crotalus horridus.     

In Vitro Cytochrome P450 2E1 and 2A Activities in the Presence of Testicular Steroids

Cytochrome P450 2E1 (CYP2E1) and 2A (CYP2A) are the main enzymes involved in the metabolism of skatole in pigs. In this study, physiological concentrations of androstenone, 17β‐oestradiol and testosterone were tested for their ability to regulate CYP2E1 and CYP2A activity in liver microsomes isolated from entire male and female pigs as well as in microsomes from Saccharomyces cerevisiae expressing either human recombinant CYP2E1 or CYP2A6. We found that physiological concentrations of androstenone and oestradiol had the ability to inhibit CYP2E1 activity. The magnitude of this inhibition (approximately 30%) was similar in recombinant human CYP2E1 and microsomes from entire male pigs. This inhibition was only seen when adding the steroid to the assay 15 min before the substrate. Interestingly, CYP2E1 activity in the microsomes from female pigs was not affected. None of the investigated steroids modified the activity of recombinant human CYP2A6. However, CYP2A activity was slightly increased in the microsomes from female pigs in the presence of oestradiol, but the magnitude of this increase was very low (below 10%) and probably irrelevant. Overall, these results indicate that physiological concentrations of androstenone and oestradiol have a potential to inhibit CYP2E1 activities in vitro, and that this inhibition is gender‐specific. Further studies are needed to investigate the biochemical mechanisms underlying those differences between the genders.     

Relationship between Oxygen Consumption and Sex of Bovine In Vitro Fertilized Embryos

The present study was conducted to examine the relationship between the oxygen consumption rate and sex ratio of bovine in vitro fertilized embryos on each day of blastocyst formation. The quality of blastocysts collected on day 7, 8, and 9 after in vitro fertilization (IVF) were categorized as ranks A and B (excellent and good, respectively) based on microscopic observation of the morphology. The oxygen consumption rate and sex of individual blastocysts were evaluated using two novel techniques: scanning electrochemical microscopy (SECM) and loop-mediated isothermal amplification (LAMP), respectively. The oxygen consumption rates of embryos of rank A were significantly higher (p < 0.05) than those of rank B, irrespective of the day of blastocyst appearance after IVF. Neither did the proportion of male embryos of ranks A and B differ significantly from each other at any of the days examined, nor from the average proportion (53%). The oxygen consumption rate of embryos of rank B collected on day 8 was significantly higher (p < 0.05) in female embryos than in male embryos collected on the same day. However, there were no apparent differences of oxygen consumption rates at each day of blastocyst appearance between male and female embryos of rank A. These results indicate that the oxygen consumption rate of individual embryos reflects their quality but does not correlate with the sex ratio of embryos of excellent quality.     

Hydrallantois in the Mare – A Report of Five Cases

Hydrallantois in the mare is a very rare condition, and clinical reports help to gather information to elucidate its pathogenesis, treatment options and prognosis. Five different cases of hydrallantois in the mare are reported in this article, all with the involvement of placentitis. The five mares were presented because of acute distention of the abdomen, dyspnoea, stiff gait and a lack of appetite. After a gradual release of the excessive amount of allantoic fluid, an abortion was induced in all five mares. The foals were either born dead or euthanized. The mares recovered quickly. One mare conceived within the same season, one remained barren despite several cycles of natural breeding, and no data were available on the other three mares. In this series, the condition is reported for the first time in two Shetland ponies, both pregnant with foals sharing a close genetic background. In both cases, the condition led to hyperlipidemia. The condition as it occurs in nulliparous mares is also discussed. Finally, the possible involvement of placentitis in the pathogenesis is emphasized.     

Effect of Transgene Introduction and Recloning on Efficiency of Porcine Transgenic Cloned Embryo Production In Vitro

Retrovirus-mediated exogenous gene transfection of somatic cells is an efficient method to produce transgenic embryos by somatic cell nuclear transfer (SCNT). This study evaluated whether efficiency of transgenic embryos production, by SCNT using fibroblast cells transfected by retrovirus vector, is influenced by the introduced transgene and whether recloning could further improve its efficiency. Transgenic cloned embryos were produced by SCNT of porcine foetal fibroblast cells transfected by either LNβ-Z or LNβ-enhanced green fluorescent protein (EGFP) retrovirus vector and evaluated for their developmental ability in vitro . Blastomeres from four-cell stage porcine embryos, produced by SCNT of foetal fibroblast cells transfected with LNβ-EGFP retroviral vector, were subsequently recloned into enucleated metaphase II oocytes and evaluated for changes in chromatin configuration, in vitro embryo development and gene expression. Analysis of results showed that cleavage and blastocyst rates of porcine SCNT embryos, using LacZ (53.6 ± 6.4%; 12.0 ± 5.7%) or EGFP (57.5 ± 6.3%; 10.1 ± 4.1%) transfected fibroblasts, did not differ (p > 0.05) from those of non-transfected controls (60.9 ± 8.2%; 12.3 ± 4.0%). Recloning of blastomeres did not further improve the in vitro development rate. Interestingly, the nuclei of blastomere underwent slower remodelling process than somatic cell nuclei. Both cloned and recloned embryos showed 100% transgene expression and there were no evidence of mosaicism. In conclusion, our data shows that the efficiency of transgenic cloned embryos production by SCNT of somatic cells transfected with replication-defective retrovirus vector is not influenced by the transgene introduction into donor cells and recloning of four-cell stage blastomere could not further improve its efficiency.