Serial Transrectal Ultrasonography for Monitoring the Reproductive Activity of the Asiatic Black Bear (Ursus thibetanus ussuricus)

This study evaluated the structural changes in the reproductive tract of Asiatic black bears using serial transrectal ultrasonography. In addition, the ultrasonographic observations were compared with the results of vaginal cytology and hormonal analyses. The collection of blood for hormonal analysis, vaginal cytology and transrectal ultrasonography was performed in two bears (Bears 1 and 2) from June 2011 to August 2013 without mating and in a third bear (Bear 3) from April to December 2012, allowing natural mating. Serial ultrasonographic observations showed cyclic changes in ovarian structures (e.g. emergence of small follicles, growth and ovulation of dominant follicles and corpus luteum (CL) formation) during the reproductive cycles of the three bears. The diameter of the uterine horns remained similar throughout the reproductive cycle in Bears 1 and 2, and it remained similar from April until October, but an enlargement containing foetuses was observed in Bear 3 in December. The ultrasonographic observations were consistent with the data obtained through vaginal cytology and progesterone analysis during the reproductive cycle. An average of 4.0 (±0.4) dominant follicles was observed during the oestrous stage (May‐August), during which the superficial cells accounted for >90% of the total vaginal cells. In addition, the detection of an average of 2.6 (±0.2) CL was associated with increased plasma progesterone concentrations (3.0 ± 0.4 ng/ml) between June and December (near hibernation). In conclusion, serial transrectal ultrasonography demonstrated yearly oestrous (ovulation) cycles via follicular dynamics and CL formation on ovaries, accordingly with vaginal cytology and hormonal level in the Asiatic black bear.     

Motility parameters of perch spermatozoa (Perca fluviatilis L.) during short-term storage with antioxidants addition

In a natural environment, seminal plasma provides spermatozoa with protection against reactive oxygen species. Storing semen in cooling conditions requires diluting it with various buffer solutions. Therefore, the protective role of seminal plasma is not sufficient enough. Semen obtained from five male specimens was diluted with the Kobayashi buffer solution at a 1:9 ratio. To determine the influence of antioxidants on semen storage, a buffer solution was used, as before, with the addition of 1 % albumin, 1 mM vitamin C, 1.5 mg ml^?1 vitamin E, 5 mM sodium citrate, 5 mM glutathione and 5 mM cysteine. After the preparation of such tests, the parameters of spermatozoa motility were measured every 3–5 days, using the CASA system (Image House CRISMAS Company Ltd.). Among all used antioxidants, the best effects were observed after the addition of glutathione to semen. After 17 days of storage, the percentage of motile spermatozoa in the samples preserved with glutathione addition was 57 %, while without antioxidant addition, it was 44 %. Furthermore, the addition of cysteine and albumin also resulted in the lengthening of the life span of perch sperm cells. The presence of the remaining antioxidants (vitamins C and E, and sodium citrate) did not have any positive influence on spermatozoa viability, and in these samples, no motile spermatozoa were observed after 12 days of storage. Our data show that dilution of perch sperm with buffered solution might be a promising method for short-term storage.     

Dietary bee pollen affects hepatic–intestinal histomorphometry of Nile tilapia fingerlings

The bee pollen is considered an excellent source of flavonoids, carotenoids, phenolic compounds, sterols and minerals; and possesses the ability to boost the immune system, antioxidant action and other interesting therapeutic effects. This study was carried out aiming to evaluate the inclusion of bee pollen in extruded commercial diets of Nile tilapia fingerlings and its effects on the hepatic‐intestinal histomorphometry and zootechnical performance. A total of 225 tilapia fingerlings (1.25 ± 0.05 g) were distributed in a completely randomized design in 15 tanks (30 L) maintained in a recirculation water system with three treatments (0% or control, 1.5% and 2.5% of bee pollen inclusion) and five replicates. Feeding rates were defined from the weekly biometrics and periodic monitoring of the physical–chemical water quality parameters. The water quality variables remained within the appropriate range for the species throughout the experiment. There was no significant difference for the somatic indexes and zootechnical parameters in this experiment. However, the inclusion of bee pollen in Nile tilapia fingerlings diets showed a linear increase in hepatocyte morphology (p = .0098). For the intestinal variables of villus height a significant linear increase was observed (p < .05) as the pollen inclusion increased. In fish that received 2.5%, the number of goblet cells was significantly higher (p < .001) than control group and 1.5%. In this sense, the inclusion up to 2.5% bee pollen in extruded commercial diets of Nile tilapia fingerlings had a positive impact on hepato‐intestinal histomorphometry without causing negative effects on the zootechnical performance.     

Role of mycorrhiza and soil fungi in natural regeneration of fir (Abies alba Mill.) in Polish Carpathians and Sudetes

Changes occurring in mycotrophy of fir seedlings (Abies alba Mill.) and in communities of the soil fungi, fungi living in roots, and fungi from rhizosphere were studied in mountain forest stands, both with good and poor regeneration of fir. The intent of these studies was to contribute to the elucidation of the problem of whether the lack of fir regeneration could be connected with microbiological changes taking place in the soil environment.     

Adjustment of annual NEE and ET for the open-path IRGA self-heating correction: Magnitude and approximation over a range of climate

The self-heating correction is known to modify open-path eddy covariance estimates of net ecosystem CO₂ exchange, typically towards reduced uptake or enhanced emissions, but with a magnitude heretofore not generally documented. We assess the magnitude of this correction to be of order 1 μmol m⁻² s⁻¹ (daytime) for half-hourly fluxes and consistently over 100 g C m⁻² for annual integrations, across a tower network (CARBORED-ES) spanning climate zones from Mediterranean temperate to cool alpine. We furthermore examine the sensitivity of the correction to its determining factors. Due to significant diurnal variation, the means of discriminating day versus night can lead to differences of up to several tens of g C m⁻² year⁻¹. Since its principal determinants - temperature and wind speed - do not include gas flux data, the annual correction can be estimated using only meteorological data so as to avoid uncertainties introduced when filling gaps in flux data. For fast retro-correction of annual integrations published prior to the recognition of this instrument surface heating effect, the annual impact can be roughly approximated to within 12 g C m⁻² year⁻¹ by a linear function of mean annual temperature. These determinations highlight the need for the flux community to reach a consensus regarding the need for and the specific form of this correction.     

Studies on the Intracellular Ca2+ Concentration of Frozen–Thawed Dog Spermatozoa: Influence of Equex from Different Sources, Two Thawing Diluents and Post-thaw Incubation in Capacitating Conditions

The addition of 0.5% (v/v) of Equex STM Paste (Nova Chemical Sales, Scituate Inc., MA, USA), whose active ingredient is sodium dodecyl sulphate (SDS), to a Tris–egg yolk extender was demonstrated to improve the longevity of frozen–thawed dog spermatozoa during in vitro incubation at 38°C. The aim of the first experiment was to compare the effects of two SDS‐containing compounds, Equex STM Paste and Equex Pasta (Minitüb, Tiefenbach, Germany), when added to a Tris–egg yolk based extender, on the post‐thaw longevity of dog spermatozoa, as well as on the intracellular Ca²⁺ concentration of spermatozoa, during post‐thaw incubation at 38°C. The post‐thaw sperm survival and longevity, as well as the quality of the sperm movement, were significantly better when using Equex STM Paste. Such prolonged sperm longevity, however, was associated to a higher intracellular Ca²⁺ concentration in a large subpopulation of the live spermatozoa. A second experiment was aimed to evaluate the effects of sperm dilution immediately post‐thaw with a Tris buffer containing glucose or fructose. The two Tris buffers were no different for any of the sperm parameters studied. The aim of a third experiment was to evaluate the sperm longevity, motility patterns and intracellular Ca²⁺ concentration of cryopreserved dog spermatozoa during post‐thaw incubation in capacitating conditions [canine capacitating medium (CCM) with or without 5 μg/ml of heparin]. Heparin had no significant effects on any of the sperm parameters evaluated. During the first 8 h of incubation, the majority of the live spermatozoa had a high intracellular Ca²⁺ content. However, after 8–10 h of incubation, it had significantly declined. The highest proportion of fast motile sperm, and the highest curvilinear velocity, average path velocity and amplitude of lateral head displacement for the total motile sperm were observed during the 2–4‐h incubation period. It was concluded that: (a) the addition of 0.5% (v/v) of Equex STM Paste to a Tris–egg yolk based extender significantly improved the post‐thaw longevity of dog spermatozoa, but the same concentration of Equex Pasta had no significant beneficial effects; (b) sperm dilution after thawing with a Tris buffer containing glucose or fructose made no difference in post‐thaw sperm longevity; (c) the addition of 5 μg/ml of heparin to CCM had no significant capacitating effects on frozen–thawed dog spermatozoa.     

DNA Status on Thawed Semen from Fighting Bull: A Comparison Between the SCD and the SCSA Tests

The assessment of sperm chromatin status is compulsory in a complete spermiogram. Here we applied the sperm chromatin structure assay (SCSA) and the sperm chromatin dispersion (SCD) test to assess the chromatin status of three fighting bulls. Cryopreserved semen (two straws/bull) were analysed by duplicate after thawing and after 6 h at 37°C with and without oxidative stress (1 m m FE²⁺). Results (SCD: percentage of spermatozoa with halo; SCSA: SD-DFI, %DFI and HDS) were analysed for differences between bulls and treatments, sensitivity and specificity (receiver operating characteristic curves) and repeatability (repeatability coefficients as 2SD of duplicate differences).%DFI for the three bulls was below 2% at 0 h, indicating no risk for fertility according to previous reports. It increased slightly for two of the bulls after FE²⁺ treatment (%DFI < 5%) and more pronouncedly for the other bull (C, %DFI∼10%), which merits further investigation. SCD rendered higher percentage of halos for bull C, but could not discriminate between samples with and without oxidizing treatment (AUC: 0.52). SCSA (%DFI) showed a high discriminating ability between treatments (AUC: 0.96). The repeatability coefficient was also higher for SCD (5.9) than for %DFI (1.8), indicating lower repeatability for SCD. Overall, %DFI might be the most useful parameter for assessing sperm chromatin on fighting bull. SCD might yield different information than SCSA, hence further research is warranted.