Effect of zearalenone on circulating testosterone concentration, testicular and epididymal morphology and epididymal sperm characteristics in wild boars.

This study investigated the effect of exposure to zearalenone (ZEN) and its metabolites on the characteristics of epididymal spermatozoa, testicular and epididymal biometry and histology, and the concentration of testosterone in blood plasma in male wild boars. The study was performed during more than one year on 18 clinically healthy male wild boars with initial and final body weight, of 39 ± 4 kg and 59 ± 3 kg, respectively. The animals were divided into two experimental groups (group I and group II) and one control group (group C) comprising 6 boars per group. Group I animals were administered per os pure zearalenone (ZEN) at 150 µg/kg BW for 7 consecutive days every two months, while group II animals received a dose of 50 µg/kg BW/day via feed that was naturally contaminated with ZEN. These male wild boars were exposed to ZEN over a period of 1 year. Control animals were fed a placebo. Testicles with epididymides of the boars were collected on the last day of the experiment within 3 min after slaughter. Blood samples were collected from each of the male wild boars. Testes and epididymides were measured and sampled for histological examination. Epididymides were dissected and epididymal spermatozoa were harvested. The spermatozoa were diluted with swine-specific BTS extender and stored at 17°C for 144 h. Sperm motility was analyzed with CASA, and other parameters including viability, acrosome integrity, DNA fragmentation index, lipid peroxidation and apoptosis were assessed with flow cytometry. In these wild boars, per os exposure to natural sources of ZEN or a combination of ZEN and its metabolites changed the testicular interstitium and led to modification of some epididymal sperm parameters. The interstitial glands in testes of experimental group I were markedly reduced and hyperemic with evident blood stasis in small capillaries. Also in group I were single degenerating seminiferous tubules. In both groups I and II, immediately after dilution of spermatozoa with BTS remarkable decreases in motility rate as well as in progressive motility and the subpopulation of cells with rapid movement were noted compared with the control group (P < 0.05). But unexpectedly, after 24 h incubation of boar semen in the BTS diluent, these sperm properties improved and were not significantly different from the control group. Thus, exposure to ZEN has no lasting but only a temporary, reversible effect on wild boar sperm motility. There was no influence of exposure to ZEN and its metabolites on the integrity of membranes, intensity of lipid peroxidation and apoptosis or on sperm chromatin structure. This study is the first using these direct measures of sperm motility and integrity to show a redundant adverse effect of ZEN exposure on wild boar sperm characteristics. There were no effects of exposure to ZEN and its metabolites on body weight, testicular and epididymal biometry, gonadosomatic index and the concentration of testosterone in blood plasma in the male wild boars.

Identification and functional analysis of bull (Bos taurus) cauda epididymal fluid proteome.

Despite recent advances in bull epididymal fluid proteome research, significant numbers of proteins secreted to epididymal lumen remain unidentified. The objective of this study was to expand the number of identified cauda epididymal fluid proteins in bulls and to contextualize them in a broader view of their mutual interactions and involvement in biological processes and pathways, to fully elucidate the ways in which epididymal fluid proteins are involved in storage and maturation of spermatozoa in epididymis. We collected postmortem cauda epididymal fluid from 6 mature Holstein Friesian bulls. We performed the identification of proteins using 2-dimensional electrophoresis coupled with MALDI mass spectrometry. Analysis of functionality and pathway involvement of identified proteins was performed using Ingenuity Pathway Analysis software. We identified a total of 189 epididymal fluid proteins, out of which 100 were newly identified in bull epididymal fluid. We have combined our data with 2 previously performed bull epididymal fluid proteome identifications, yielding 280 proteins total, and analyzed it. The main canonical pathways involving epididymal proteins were glycolysis, gluconeogenesis, protein ubiquitination pathway, nuclear factor-erythroid 2-related factor 2-mediated oxidative stress response, and farnesoid X receptor/retinoid X receptor activation. The main biological functions potentially performed by epididymal fluid proteins included carbohydrate metabolism, cellular growth and proliferation, cell death and survival, and small molecule biochemistry. Overall, our results have pointed out multiple novel pathways in bull epididymal fluid that might take part in various aspects of maturation and protection processes of epididymal spermatozoa.

Analysis of bull (Bos taurus) seminal vesicle fluid proteome in relation to seminal plasma proteome.

The existing knowledge on the bull seminal vesicle proteome, a major seminal plasma constituent, and its relationship with seminal plasma is limited. This knowledge is prerequisite for a better understanding of seminal plasma variability, which is linked to semen quality. The objective of this study was to characterize the proteomes of seminal vesicle fluid and seminal plasma and to compare them to better understand the origin of seminal plasma proteins. We collected ejaculates and seminal vesicle fluid postmortem from 6 mature Holstein Friesian bulls. We performed the analysis and identification of proteins using 2-dimensional electrophoresis coupled with matrix-assisted laser desorption/ionization mass spectrometry. We identified 105 proteins in bull seminal vesicle fluid and 88 proteins in seminal plasma. For both seminal vesicles and seminal plasma proteins described in our study, top biological functions were cellular movement, cell death and survival, and cellular growth and proliferation. Additionally, seminal vesicle fluid proteins were involved in protein degradation and synthesis. Seminal plasma proteins were also involved in cellular assembly and organization and cell-to-cell signaling and interactions. Proteins of both fluids were involved in the following canonical pathways: glycolysis, gluconeogenesis, liver X receptor/farnesoid X receptor, and farnesoid X receptor/retinoid X receptor activation. Additionally, seminal vesicle fluid proteins appeared to be involved in oxidative stress response mediated by nuclear factor E2-related factor 2. Our results described the bull seminal vesicle fluid proteome for the first time and allowed for significant expansion of the current knowledge on the bull seminal plasma proteome. Moreover, analysis indicated that both bull seminal vesicle fluid and seminal plasma proteomes contained interconnected protein groups related to protective functions, glycolysis, and the morphology and physiology of the spermatozoa. These proteins and their interactions could be targeted in future research.

Relationships between ovarian cysts and morphological and hormonal state of ovarian cortex in sows.

The aim of this study was to evaluate the relationship between ovarian cysts and concentrations of ovarian steroid hormones: 17beta-estradiol (E(2)), progesterone (P(4)), testosterone (T), and androstendione (A(4)) both in blood plasma and in cysts and morphological state of the ovarian cortex in sows. Females were divided into three groups: PCO (sows with polycystical ovaries), OO (sows with oligocystic ovaries) and control (sows without ovarian cysts). The ovaries for evaluations were collected after slaughtering of 18 multiparous sows. Between the PCO and OO animals, statistically significant differences in numbers of the follicular cysts (FC) (8.6 vs. 1.5), follicular theca-lutein cysts (FTLC) (8.0 vs. 2.0), follicular lutein cysts (FLC) (4.5 vs. 2.0) and corpus luteum cysts (CLC) (1.7 vs. 0.4) (P< or =0.01) were noted. In the PCO sows the most common kinds of cysts were FC and FTLC (8.6 and 8.0) whilst in OO sows the cysts occurred on their ovaries on a similar level (FC - 1.6, FTLC - 2.0, FLC - 2.0). Existence of more than 10 ovarian cysts in the sows significantly decreases the frequency of physiological ovarian follicles (primary, growing and maturing) and significantly increases the pathological process of atresia on all stages of ovarian follicles development (P< or =0.01). The study did not reveal any effect of growing or decreasing number of ovarian cyst on concentrations of E(2) and P(4) in blood plasma of sows. Polycystical ovaries significantly decreased concentrations of A(4) but increased the concentration of T in blood plasma (P< or =0.01). The general presence of ovarian cysts considerably positively correlated with concentrations of E(2), T and A(4) from cysts' fluid, of all kinds of ovarian cysts and atresia of primary follicles (a correlation coefficient r from 0.72 up to 0.97, P< or =0.05). The phenomenon of ovarian cysts significantly negatively correlated with all generations of ovarian follicles (P< or =0.05).

Effects of three cryopreservation methods and two semen extenders on the quality of dog semen after thawing.

The aim of this study was to evaluate the influence of two diluents, one with and the other without Orvus ES paste, and three methods of cryopreservation on the quality of dog semen after thawing. The investigation was carried out on 126 ejaculates collected from 37 dogs. In Expt 1, sperm-rich fractions of ejaculates were frozen in 0.25 ml minitubes and pellets. In Expt 2, each sperm-rich fraction of ejaculate was divided and extended in two diluents, one with and the other without Orvus ES paste. Samples of semen were frozen in pellets and 0.5 ml French straws. Motility, percentage of spermatozoa with normal acrosomes and longevity of spermatozoa were significantly higher (P < 0.05) in semen samples frozen in pellets than in samples frozen in 0.25 ml minitubes. Aspartate aminotransferase activity in extracellular fluid was significantly (P < 0.05) lower in semen frozen in pellets. There were no significant differences in quality after thawing between semen samples frozen in pellets and 0.5 ml French straws. Longevity, unlike acrosome status and aspartate aminotransferase activity, was greater in samples extended in Tris-buffered diluent with addition of Orvus ES paste, regardless of the method of cryopreservation.