Effect of Different Manganese Concentrations during in vitro Maturation of Bovine Oocytes on DNA Integrity of Cumulus Cells and Subsequent Embryo Development

Manganese (Mn) is a trace element present in forages and cereals, and its concentration depends on soil status. Manganese deficiency in cattle, goats and ewes not only impairs oestrous cycle but reduces calf birth weight. The achievement of the first oestrus is delayed, and more attempts are necessary to obtain a successful conception. This study was conducted to investigate the effect of the availability of supplemental Mn during IVM on DNA damage of cumulus cells and total glutathione (GSH) content in oocytes and cumulus cells. The effect of supplementary Mn during IVM on subsequent embryo development was also studied. The results reported here indicate (i) DNA damage in cumulus cells decreased with 0, 2, 5 and 6 ng/ml Mn supplementation during IVM (p < 0.05). (ii) Intracellular GSH‐GSSG content increased (p < 0.01) with different Mn concentrations in oocytes and cumulus cells. Also, cumulus cell number per cumulus oocyte‐complexes (COC) did not differ either before or after IVM. (iii) Addition of Mn to maturation medium resulted in similar cleavage rates (p > 0.05) at 0, 2, 5 and 6 ng/ml Mn. However, subsequent embryo development to blastocyst stage was significantly higher (p < 0.01) in oocytes matured with 5 and 6 ng/ml Mn. (iv) There was also an increase (p < 0.05) in mean cell number per blastocyst obtained from oocytes matured with 5 and 6 ng/ml respect to zero Mn (IVM alone) and 2 ng/ml Mn. This study provides evidence that optimal embryo development to the blastocyst stage was partially dependent on the presence of Mn during IVM. Moreover, the availability of Mn during oocyte maturation ensures ‘normal’ intracellular GSH content in COCs and protects DNA integrity of cumulus cells.     

Validation of actigraphy to assess circadian organization and sleep quality in patients with advanced lung cancer

Background

Many previous studies have documented seasonal variation in suicides globally. We re-assessed the seasonal variation of suicides in Finland and tried to relate it to the seasonal variation in daylength and ambient temperature and in the discrepancy between local time and solar time.

Methods

The daily data of all suicides from 1969 to 2003 in Finland (N = 43,393) were available. The calendar year was divided into twelve periods according to the length of daylight and the routinely changing time difference between sun time and official time. The daily mean of suicide mortality was calculated for each of these periods and the 95% confidence intervals of the daily means were used to evaluate the statistical significance of the means. In addition, daily changes in sunshine hours and mean temperature were compared to the daily means of suicide mortality in two locations during these afore mentioned periods.

Results

A significant peak of the daily mean value of suicide mortality occurred in Finland between May 15th and July 25th, a period that lies symmetrically around the solstice. Concerning the suicide mortality among men in the northern location (Oulu), the peak was postponed as compared with the southern location (Helsinki). The daily variation in temperature or in sunshine did not have significant association with suicide mortality in these two locations.

Conclusions

The period with the longest length of the day associated with the increased suicide mortality. Furthermore, since the peak of suicide mortality seems to manifest later during the year in the north, some other physical or biological signals, besides the variation in daylight, may be involved. In order to have novel means for suicide prevention, the assessment of susceptibility to the circadian misalignment might help.     

Distinct Effects of Boar Seminal Plasma Fractions Exhibiting Different Protein Profiles on the Functionality of Highly Diluted Boar Spermatozoa

The aim of this study was to evaluate how different protein profiles of seminal plasma (SP) fractions affect sperm functionality in vitro. Ejaculates from three boars were separated into six fractions. The fractions differed from each other in their sperm content, in their total SP protein content, and their spermadhesin PSP-I/PSP-II and heparin-binding protein (HBP) concentrations. Spermatozoa were mainly recovered in fraction 2 (sperm-rich fraction, >1800 × 10⁶ spermatozoa/ml), whereas the pre-sperm fraction 1 and the post-sperm fractions 4–6 contained low numbers of spermatozoa (<500 × 10⁶/ml). Except in fraction 2, the total SP protein concentration and the concentration of both, spermadhesin PSP-I/PSP-II and the HBPs increased with fraction order. Distinct time-dependent effects were observed on motility characteristics and membrane integrity of highly diluted boar spermatozoa upon incubation with a 10% dilution of the SP from each fraction. The highest sperm viability was recorded after exposure for 5 h to fraction 2, followed by fractions 1 and 3. The percentages of motile spermatozoa also differed significantly among fractions after 5 h of incubation. Spermatozoa incubated with SP of fractions 1–3 showed the highest percentage motility. We conclude that different SP fractions exert distinct effects on the functionality of highly diluted boar spermatozoa. Fractions 1–3 appear to promote sperm survival, whereas fractions 4–6 seem to be harmful for preserving the physiological functions of highly diluted boar spermatozoa.     

Intratesticular lidocaine reduces the response to surgical castration in dogs

ObjectiveTo investigate whether intratesticular injection of lidocaine pre-surgery would reduce the intraoperative responses to elective castration in dogs.Study designDouble-blinded, randomized, controlled, prospective clinical study.AnimalsForty-two client-owned dogs weighing 2.2–38.4 kg and aged between 4.5 and 56 months.MethodsGroup L dogs received an intratesticular injection of 2% lidocaine (2 mg kg^?1) and Group S an identical volume of saline prior to surgery. Premedication was with acepromazine and morphine intramuscularly. Anaesthesia was induced with propofol intravenously and maintained with isoflurane vaporized in oxygen. Heart rate (HR), mean arterial pressure (MAP), respiratory rate (f_R), end-tidal isoflurane (Fe′ISO) and carbon dioxide concentrations, oxygen saturation and ECG were monitored during surgery. Fe′ISO was maintained at 1.0 ± 0.1%. Supplemental propofol was given in response to gross movement.ResultsGroup L had significantly lower maximum values for both HR and MAP. Group L displayed significantly smaller increases in HR during exteriorization of the first testis than Group S. There was an overall significant difference in MAP between groups during all surgical events (p = 0.041) and time points (p = 0.002). In univariate analysis, Group L showed significantly less changes in MAP during skin incision, exteriorization of the first testis and clamping of both spermatic cords. Group S reached its highest f_R significantly earlier. Group L (eight dogs) required additional propofol 33 ± 18 minutes after the start of surgery and Group S (seven dogs) at 19 ± 17 minutes; this difference was not statistically significant. Seven dogs in Group L and 12 dogs in Group S required rescue analgesia with morphine (GCMPS-SF score ≥6); this difference was not statistically significant. No adverse effects were reported postoperatively.Conclusions and clinical relevanceBased on this study, the authors recommend the use of intratesticular lidocaine for surgical castration in dogs.     

Failure of a single postpartum prostaglandin treatment to improve the reproductive performance of dairy cows

A study was conducted to evaluate the effects of postpartum prostaglandin treatment on reproduction in 3 seasonal calving dairy herds. Recently calved lactating dairy cows were paired on herd, age, calving date and previous production index. One cow in each of the 196 pairs received a single intramuscular injection of 25 mg of the prostaglandin analogue, dinoprost, between 14 and 28 days after calving. Subsequent reproduction was monitored. Within each herd and overall, there was no significant effect of treatment on the intervals from calving to first service, mating start date to first service, calving to conception, mating start date to conception and first service to conception. Treatment also had no significant effect on 21-day submission and pregnancy rates, on the proportion of each group not pregnant at the end of mating, and on first service pregnancy rates. Responses to treatment did not vary between cows calving within 50 days of mating start date and earlier calving cows or between cows aged less than 5 years and older cows.     

Mercury contamination of the harbour porpoise Phocoena phocoena and other cetaceans from the North Sea and the kattegat

Total Hg levels were determined in 17 harbour porpoises Phocoena phocoena collected in the North Sea and the Kattegat (muscle, liver and kidney), as well as in a sperm whale Physeter macrocephalus, a common dolphin Delphinus delphis and two bottle-nose dolphins Tursiops truncatus collected in Belgium. Juvenile porpoises (total body length less than 140 cm) were characterized by total Hg concentrations increasing with length (t.i. with age) with median values of 3.1 μg g^?1 dw in muscle, 6.2 in liver and 5.7 in kidney, and liver to muscle resp. liver to kidney ratios varying between 1 and 2. In adults, liver concentrations were higher (up to 500 μg g^?1 dw) with liver to muscle and to kidney ratios up to 20. Median McHg concentrations in 5 of the porpoises were 4.0 μg g^?1 dw in muscle and 6.9 in liver. These values did not increase as strongly with length (t.i. with age, at least for juveniles) as total Hg did; as a consequence, the relative McHg levels decreased from 100% in juveniles to 2 or 3 % in the liver of adults. Such results seem to reflect the existence of a slow mineralization phenomenon and storage of inorganic Hg, mainly in the liver.     

Serological status of cattle to bovine viral diarrhoea virus and bovine herpesvirus 1 at entry to and exit from Australian feedlot backgrounding facilities

A total of 6195 cattle were enrolled in this observational study. Serum antibody concentrations to bovine herpesvirus 1 (BHV1) and bovine viral diarrhoea virus (BVDV) were measured at entry to and exit from backgrounding facilities to assess their statuses on arrival and the extent of seroconversion to these viruses during backgrounding. The backgrounding facilities were contiguous with five feedlots in: Queensland (two sites), New South Wales, South Australia and Western Australia. Cattle were held in the backgrounding facilities for a minimum of 29 days and a median of 34 days. On backgrounding facility entry, 32.7% of the study population was seronegative to BVDV, but 85.7% was seronegative to BHV1. After commingling in the backgrounding facilities, of the cattle that were seronegative on backgrounding facility entry, 33.9% and 30.3% showed a serological increase to BVDV and BHV1, respectively. At backgrounding facility exit, when cattle were placed in their feedlots, 19.6% and 59.1% were seronegative to BVDV and BHV1, respectively, and 0.26% were persistently infected with BVDV. There was a strong association between seroincrease to BVDV and seroincrease to BHV1 (P = 0.005) at animal level in cohorts known to contain an animal persistently infected with BVDV.     

Dose–Response Studies for Pituitary and Testicular Function in Male Dogs Treated with the GnRH Superagonist, Deslorelin

We tested the effect of dose of GnRH superagonist on pituitary and testicular function in a study with four groups of four male dogs. The Controls received blank implants and the other three groups received implants containing 3, 6 or 12 mg deslorelin (d ‐Trp⁶‐Pro⁹‐des‐Gly¹⁰‐GnRH ethylamide). In all deslorelin‐treated groups, there was initially an acute increase in plasma concentrations of LH and testosterone, followed by declines such that both hormones became undetectable after approximately 12 days. There was a dose–response in some of these early aspects of the hormone profiles. With respect to long‐term effects of treatment, the 12‐mg dose had significantly greater effects than the smaller doses for the duration of minimum testicular volume [366 ± 77, mean ± SEM (3 mg), 472 ± 74 (6 mg), and 634 ± 59 (12 mg) days], absence of ejaculate [416 ± 88 (3 mg), 476 ± 83 (6 mg), and 644 ± 67 (12 mg) days], undetectable plasma concentrations of LH and testosterone [367 ± 64 (3 mg), 419 ± 72 (6 mg), and 607 ± 69 (12 mg) days], the delay until complete recovery of LH and testosterone secretion [394 ± 65 (3 mg), 484 ± 72 (6 mg) and 668 ± 47 (12 mg) days], and the delay until testes had regrown to normal volume [408 ± 77 (3 mg), 514 ± 74 (6 mg), 676 ± 59 (12 mg) days]. The time taken to restore full ejaculates was also longest for the 12‐mg dose: 716 ± 67 (12 mg) days vs 440 ± 66 (3 mg) and 538 ± 83 (6 mg) days after implantation. There was no correlation between delay to recovery of normal ejaculate quality and body mass. We conclude that the dose–response relationship with deslorelin implants is not expressed with respect to the degree of suppression of reproduction, but on the maximum duration of suppression and thus to delay until recovery.