Response of Thawed Epidi dymal Red Deer Spermatozoa to Increasing Concentrations of Hydrogen Peroxide,and Importance of Individual Male Variability

Oxidative stress represents a challenge during sperm manipulation. We have tested the effect of increasing hydrogen peroxide (H₂O₂) levels on red deer spermatozoa after cryopreservation, and the role of male‐to‐male variation in that response. In a first experiment, eight thawed samples were submitted to 0, 25, 50, 100 and 200 μm H₂O₂ for 2 h at 37°C. Intracellular reactive oxygen species (H₂DCFDA‐CM) increased with H₂O₂ concentration, but we only detected a decrease in sperm function (motility by CASA and chromatin damage by sperm chromatin structure assay) with 200 μm . Lipoperoxidation assessed by the thiobarbituric acid reactive substance (TBARS) method increased slightly with 50 μm H₂O₂ and above. In a second experiment, samples from seven males were submitted to 0 and 200 μm H₂O₂ for 2 h, triplicating the experiment within each male. Males differed at thawing and regarding their response to incubation and H₂O₂ presence. We found that the kinematic parameters reflected male‐to‐male variability, whereas the response of the different males was similar for lipid peroxidation and viability. A multiparametric analysis showed that males grouped differently if samples were assessed after thawing, after incubation without H₂O₂ or after incubation with H₂O₂. Red deer spermatozoa are relatively resilient to H₂O₂ after thawing, but it seems to be a great male‐to‐male variability regarding the response to oxidative stress. The acknowledgement of this individual variability might improve the development of optimized sperm work protocols.     

Historical patterns of fire severity and forest structure and composition in a landscape structured by frequent large fires: Pumice Plateau ecoregion,Oregon, USA

Context

Lack of quantitative observations of extent, frequency, and severity of large historical fires constrains awareness of departure of contemporary conditions from those that demonstrated resistance and resilience to frequent fire and recurring drought.

Objectives

Compare historical and contemporary fire and forest conditions for a dry forest landscape with few barriers to fire spread.

Methods

Quantify differences in (1) historical (1700–1918) and contemporary (1985–2015) fire extent, fire rotation, and stand-replacing fire and (2) historical (1914–1924) and contemporary (2012) forest structure and composition. Data include 85,750-ha tree-ring reconstruction of fire frequency and extent; >?375,000-ha timber inventory following >?78,900-ha fires in 1918; and remotely-sensed maps of contemporary fire effects and forest conditions.

Results

Historically, fires?>?20,000 ha occurred every 9.5 years; fire rotation was 14.9 years; seven fires?>?40,469 ha occurred during extreme drought (PDSI <?? 4.0); and stand-replacing fire occurred primarily in lodgepole (Pinus contorta var. murrayana). In contemporary fires, only 5% of the ecoregion burned in 30 years, and stand-replacing fire occurred primarily in ponderosa (Pinus ponderosa) and mixed-conifer. Historically, density of conifers?>?15 cm dbh exceeded 120 trees/ha on?<?5% of the area compared to 95% currently.

Conclusions

Frequent, large, low-severity fires historically maintained open-canopy ponderosa and mixed-conifer forests in which large fire- and drought-tolerant trees were prevalent. Stand-replacing patches in ponderosa and mixed-conifer were rare, even in fires >?40,469 ha (minimum size of contemporary “megafires”) during extreme drought. In this frequent-fire landscape, mixed-severity fire historically influenced lodgepole and adjacent forests. Lack of large, frequent, low-severity fires degrades contemporary forest ecosystems.

     

Flow and storage in groundwater systems

The dynamic nature of groundwater is not readily apparent, except where discharge is focused at springs or where recharge enters sinkholes. Yet groundwater flow and storage are continually changing in response to human and climatic stresses. Wise development of groundwater resources requires a more complete understanding of these changes in flow and storage and of their effects on the terrestrial environment and on numerous surface-water features and their biota.     

A survey of free-living ixodid ticks on a commercial game farm in the Thabazimbi District, Limpopo Province, South Africa

Free-living stages of ticks on a commercial game farm in the Thabazimbi District, Limpopo Province, South Africa, were collected by drag-sampling with flannel strips during the period September 2003 to August 2004. A total of 5 tick species was collected from 4 sites. Boophilus decoloratus was the most abundant species, followed by Amblyomma hebraeum. Seasonal abundance of the ticks was quantified and an optimum time to implement control measures against the ticks is proposed.     

疑似碘酊致牛死亡一例

笔者在临床上曾遇到一疑似“碘酊”致牛死亡的病例,由于未曾见过碘酊致牛死亡的报道,查阅资料也终未得到答案.笔者将其整理如下,供同行探讨。     

Estriolum Treatment in the Bitch: A Risk for Uterine Infection?

Purulent vaginal discharge in a bitch in which ovariohysterectomy has been performed is often caused by inflammation of the uterine stump. The inflammation is due to either cystic endometrial hyperplasia (CEH) induced primarily by progesterone from remnant ovarian tissue or exogenous progestagens, or it is due to the presence of unabsorbed suture material. This report describes a 9-year-old Irish setter with hemopurulent vaginal discharge and non-pruritic symmetrical alopecia, which had undergone ovariohysterectomy 3.5 years ago and which had been treated with estriolum daily for the past 2.5 years because of urinary incontinence. Vaginoscopy revealed hemopurulent discharge throughout the vagina and vestibule. Cytological examination of ultrasound-guided fine-needle aspiration biopsies of a large mass in the hypogastricum, which appeared to be the uterine cervical stump, revealed septic purulent inflammation. The concentration of plasma progesterone was low and the concentration of plasma 17-ß oestradiol did not increase after gonadotrophin-releasing hormone administration. No remnant ovarian tissue was found by abdominal ultrasonography, laparotomy, or histological examination of mesovarian pedicles. Laparotomy revealed uterine stump empyema. Histological examination of the surgically removed mass excluded both CEH and unabsorbed suture material as the cause of the stump empyema. Instead, it is hypothesized that the long-term treatment with estriolum was a causative factor. This suggests that bitches treated with estriolum should be examined regularly.     

Percentage of Ubiquitinated Spermatozoa does not Correlate with Fertilizing Capacity of Thawed Bovine Semen

In the spermatozoa of some species, the ubiquitin–proteasome system detects altered proteins and tags them for elimination by the proteasome. In some species' ejaculates, a high proportion of ubiquitinated spermatozoa (i.e. those having ubiquitin bound to the altered or damaged membrane proteins) has been related to infertility. The aim of this study was to assess whether the percentage of ubiquitinated spermatozoa relates to fertility of dairy bulls and whether ubiquitination increases during protein remodelling that occurs during in vitro spermatic capacitation. Thirty‐two frozen semen straws from four high‐fertility (ReproMax^®) and four normal‐fertility (Normal) Holstein‐Friesian sires were evaluated. Ubiquitinated and capacitated spermatozoa were quantified by sperm ubiquitin tag immunoassay and chlortetracycline stain, respectively. Fertilizing capacity of sires was assessed by in vitro fertilization. No differences were found between Normal and ReproMax^® sires with regard to the observed percentage of ubiquitinated spermatozoa (42.97 ± 3.69% and 49.68 ± 9.27%, respectively; p > 0.05). Additionally, no differences were found in the percentage of ubiquitinated spermatozoa as a consequence of spermatic capacitation in either Normal (42.97 ± 3.69% before capacitation vs 44.67 ± 7.5% after; p > 0.05) or ReproMax^® sires (49.68 ± 9.27% before vs 45.05 ± 7.51% after; p > 0.05). The percentage of ubiquitinated spermatozoa in a thawed sperm samples did not correlate with its in vitro fertilizing capacity; thus, this assay does not prove useful to detect in vivo fertility differences between sires. Additionally, protein degradation occurring during remodelling of the spermatozoon plasma membrane during the capacitation process does not seem to involve the ubiquitin–proteasome system.