Effect of dietary supplementation with seaweed and polyphenols mixture on antioxidant status, concentration and motility of rabbit spermatozoa

In recent years, many studies have been focused on natural substances that could have effect on health of animals. We investigated effect of extract consisting mainly of polyphenols, brown algae and plant polysaccharides on the reproduction potential of male rabbits (New Zealand white breed) during 60 days long dietary experiment. The rabbits were divided into three groups. Control was fed a basal diet, whereas the second and third group were supplemented with seaweed and polyphenols mixture: T1 = 0.3% and T2 = 0.6% respectively. We observed that sperm concentration in both experimental groups increased in comparison to the control group. Results of the CASA analysis showed enhanced motility (C = 85.09±7.53%, T1 = 87.21±8.25%, T2 = 89.38±8.02%) and progressive motility (C = 74.28±12.6%, T1 = 79.07±13.89%, T2 = 81.28±11.37%) in experimental groups supplemented with combination of algae and polyphenols in comparison with the control group. While monitoring ferric reducing ability of plasma (FRAP), we found the highest value in T1 group, changes were insignificat. An increase in GPx activity was measured in experimental groups in comparison with the control group with major difference in T1 group. In experimental groups, we determined an increase in activity of superoxide dismutase (SOD) in comparison with control group, the difference was significant in T2 group (C = 0.26±0.11 U/mg TP; T2 = 0.31±0.09 U/mg TP). In conclusion, our studies suggest that dietary supplementation with brown seaweed and plant polyphenols mixture may be potentially useful for enhancement of sperm motility and protection against oxidative stress

Lippia citriodora (verbascoside) extract supplementation: Effect on rabbit semen quality in vivo and in vitro

Verbascoside, the main component of Lippia citriodora extract, is one of the most powerful free radical scavengers exhibiting a wide biological activity. In in vivo study 20 adult New Zealand white rabbit bucks were divided into two homogeneous groups, one control (CON) and one verbascoside-supplemented (0.1%) in feed mixture (EXP) and later in vitro effects of verbascoside on the motility aspects of rabbit spermatozoa were analysed. The spermatozoa concentration, ejaculate volume, spermatozoa motility, progressive motility, distance parameters, velocity parameters and type of spermatozoa movement were negatively affected by Lippia citriodora leaves extract after the first 4 weeks of dietary treatment, till the end of experiment (8 weeks). Four weeks after the suspension of feed additive supplementation, all spermatozoa traits values returned to the normality, and in line with CON group. For in vitro findings, ejaculates from 10 male New Zealand white bucks were collected using an artificial vagina. Then it was diluted in physiological saline solution containing different concentrations of verbascoside at the concentration of 0, 0.0024, 0.0219, 0.157, 120.0 mg/ml (Ctrl, VB1, VB2, VB3, VB4 groups, respectively), using a dilution ratio of 1: 4. The obtained data proved that verbascoside at the concentration of 0.0024 and 0.0219 mg/ml had no adverse effect on spermatozoa. Additionally, we found that verbascoside at higher concentrations (0.157 and 120.0 mg/ml) significantly altered all the motility parameters analysed in the experiment. In conclusion a possible negative effect of verbascoside supplementation into feed mixture (0.1%) on semen quality parameters in rabbit bucks as well as in vitro can be stated, obviously considering that target organs of antioxidant activities of phenylpropanoid glycosides are various. In addition it has to be emphasized that the extract showed a reversible action, since the semen traits of treated animals returned to the normality after the dietary administration period

Effects of dietary plant polyphenols and seaweed extract mixture on male-rabbit semen: Quality traits and antioxidant markers

Feed additive consisting of polysaccharides from brown seaweeds plus phenolic acid, hydroxycinnamic acids, tannins, and flavonoids from plant extracts, was used as rabbit food supplement. Main aim of the study was to determine the effect of natural mix of marine and freshwater seaweed polyphenols on selected reproductive characteristics of male rabbits during the 90-days experiment. Natural mix was incorporated in feed-pellets for rabbits in two different concentrations – 0.3% (T1 group) and 0.6% (T2 group), compared with a control group (C group). In experimental groups a significant increase of concentration of calcium during first 30 days of supplementation was found. An increase of alanine aminotransferase, glutathione peroxidase and ferric reducing ability of plasma and a decrease in aspartate aminotransferase after 90 days were recorded in the same groups. Except for that we noticed decrease of semen distance of curved line and velocity of curved line after 30 days though only while being supplemented with 0.6% proportion of seaweed polyphenols in feed mixture. Based on the results it can be stated that the natural mix in the tested levels do not show adverse effect on male rabbit reproductive parameters, and an improvement of antioxidant status was observed. The feed additives can have a very important effect on growth, health and development of animals in general as it supplies the with the much-needed minerals, nutritional substances and antioxidants, on which we focused in our study

Physiological and Pathological Roles of Free Radicals in Male Reproduction

Oxidative stress (OS) is a condition caused by an imbalance between reactive oxygen species (ROS) overgeneration and decreased antioxidant defense mechanisms in the cell. OS has become a prominent factor in male reproductive dysfunction as ROS cause damage to sperm DNA, lipids and proteins, alterations to critical sperm structures and signaling pathways, leading to a decreased sperm activity and fertilizing capacity. At the same time, small amounts of ROS play vital roles in events leading to sperm maturation and acquisition of functional activity, which is why a proper oxidative balance is of paramount importance for a proper male fertility. Understanding the physiological and pathological roles of ROS in male reproduction has become an essential pillar of modern andrology; however, numerous questions related to the controversial behavior of ROS in male reproductive cells and tissues still remain unanswered. This chapter aims to summarize current evidence available on the relationships between free radicals, antioxidants and male reproduction and to trigger more scientific interest, particularly with respect to the design of efficient strategies to diagnose or treat male sub- or infertility associated with OS

Comparison of semen characteristics and histological structure of the testis from transgenic and non-transgenic rabbits

[EN] The aim of this study was to compare semen characteristics including sperm quantity, quality, and abnormalities, as well as histological structure of the testis of three-year old transgenic (human clotting factor, hFVIII, gene) and nontransgenic rabbits. For the experiment, 10 transgenic rabbits of F2 and F3 generations and 10 randomly selected non-transgenic males of the same breed and age were used as controls. All males were housed in individual cages, under a the same environmental conditions: photoperiod (14L:10D), temperature (18-20°C), and humidity (65-70%). Semen samples, collected once a week for 20 wk from each control and transgenic male, were analyzed by computer assited semen analysis within a few minutes following natural ejaculation into an artificial vagina. Concentration of spermatozoa was higher in the transgenic than in the non-transgenic group (P<0.001; 316.6±148.8 and 126.7±64.4¿106/ mL, respectively). Significant differences (P<0.1) between transgenic and non-transgenic males were observed also in spermatozoa motility (63.08 vs. 32.60%). Significantly higher (P<0.05) relative volume (8.08±2.89%) and diameter of testicular lumen (36.89±23.11 ¿m) were found in the transgenic animals compared to control animals (16.69±4.70%, 53.89±25.42 ¿m). Our results show that spermatozoa parameters and histological structure of the testis can be used for the characterization of male reproductive traits of older transgenic rabbits.This work was supported by the grant No: 2003 SP51/028 09 00/028 09 03 coordinated by the Slovak Academy of Science and by the grant of Ministry of Agriculture Slovak Republic (RÚVV 07-012). All experiments were approved according to ethical permission No. SK P 28004. We are grateful to Dr. Shoubadeep Roychoudhury for the English correction.Lukac, N.; Massanyi, P.; Flesarova, S.; Danko, J.; Makarevich, A.; Chrenek, P. (2009). Comparison of semen characteristics and histological structure of the testis from transgenic and non-transgenic rabbits. World Rabbit Science. 17(4):221-226. https://doi.org/10.4995/wrs.2009.64722122617

The effects of caffeine on the motility and viability of stallion spermatozoa at different temperature conditions

The purpose of this study was to evaluate the dose-and time-dependent effect of caffeine treatment on the motility and viability of stallion spermatozoa at different temperatures. Six dose groups (A to F) were established with changing caffeine concentrations (from 0.625 to 10 mg/mL). The control samples were prepared by diluting the ejaculate only with physiological salt solution. The samples were examined after 0, 1, 2 and 3 h of incubation at 5 degrees C and 37 degrees C. The motility parameters were evaluated by Computer Assisted Semen Analyzer (CASA) system, and the viability was assessed by the mito-chondrial toxicity test at the end of the incubation. A positive effect of the lowest tested caffeine concentration on the motility parameters was observed throughout the incubation period at 5 degrees C. At the end of the 3h incubation, the viability in every sample in these groups, treated with any caffeine concentration, showed lower values compared to the control. At the higher incubation temperature (37 degrees C), caffeine positively affected the motility in samples B (P < 0.05) and D, E, F (P < 0.001) after 3 h of incubation; however, the viability showed a slightly decreasing tendency. Our results suggest that caffeine, in an optimal concentration, may be used as a component of stallion semen extenders

Endocrine Disruption by Heavy Metals on Steroidogenesis in Model Systems

In this study human adrenocortical carcinoma cell line NCI-H295R was used as an in vitro biological model to study the effect of heavy metals on steroidogenesis. The cell cultures were exposed to different concentrations of cadmium (1.90; 3.90; 7.80; 15.60; 31.20; 62.50 μM of CdCl2), mercury (1.0; 5.0; 25; 50; 100 μM of HgCl2), nickel (3.90; 7.80; 15.60; 31.20; 62.50; 125; 250; 500 μM of NiCl2) and compared to control. Cell viability was measured by the metabolic activity (MTT) assay for estimation of mitochondria structural integrity. Quantification of sexual steroid production directly from aliquots of the medium was performed by enzyme linked immunosorbent assay (ELISA). Cadmium decreased the release of progesterone and testosterone already at a very low concentration (1.90 μM) of CdCl2, while the cell viability remained relatively high (> 75%) up to 7.80 μM of CdCl2 and significantly (P<0.01) decreased at 15.60 μM and higher concentrations of CdCl2. Concentration-dependent depression in testosterone production was detected particularly for higher concentration of HgCl2. Progesterone production was also decreased, but at the lower concentrations (1.0 and 5.0 μM) of HgCl2 this decline was lower compared to depression of testosterone. The cell viability significantly decreased at 25 μM and higher concentration of HgCl2. Results of the our study indicate dose dependent decreases in both sexual steroid hormones by NCI-H295R cell culture following a 48 h in vitro NiCl2 exposure. The lowest concentration of progesterone was significantly (P<0.01) detected in groups with the higher doses (≥ 500 μM) of NiCl2, which elicited significant cytotoxic effect. The testosterone production was decreased as well, but this decline was more pronounced compared to depression of progesterone. These results suggest that heavy metals have detrimental effects on steroid hormone synthesis even at very low concentrations and consecutively on reproductive physiology

EFFECT OF ZEOLITE (CLINOPTILOLITE) ON CHEMICAL PARAMETERS OF RABBIT MEAT

This work is aimed to evaluate the effect of natural zeolite on selected parameters of the rabbit's meat chemical composition. The rabbits of the Castorex breed (n=12) were divided into two groups: Control group C (n=6) without the addition of zeolite in feed and the experimental group Z (n=6) in which animals received zeolite daily in a peroral form in concentration 0.2 g/kg of body weight. Determination of the nutritional composition of rabbit meat and analysis of individual amino acids and fatty acids was performed by the Nicolet 6700 using FT-IR method (fourier transform infrared spectroscopy) in the musculus Longissimus dorsi (MLD) and the musculus Vastus lateralis (MVL). The content of water in meat in group Z (73.630 ± 0.270 g * 100g-1) was significantly higher (P &lt; 0.05) compared to group C (72.480 ± 0.530 g * 100g-1). Cysteine content in MLD in group Z (0.289 ± 0.007 g*100 g-1) was significantly (P &lt; 0.05) higher than in group C (0.277 ± 0.011 g*100 g-1). Another noticeable difference in the statistical level p &lt; 0.05 in favor of zeolite was found in linoleic acid content in MLD with mean values in group Z (0.324 ± 0.016 g*100g-1 FAME) against the group C (0.293 ± 0.009 g* 100 g-1 FAME). Therefore, the addition of zeolite to the rabbit feeding diet possibly increases the proportion of essential linoleic fatty acid and thereby may improve the nutritional value of the meat. Increased cysteine value can lead to a better degradation of heavy metals in meat

Correlated Response to Selection for Litter Size Residual Variability in Rabbits' Body Condition

[EN] Selection for decreasing litter size residual variance has been proposed as an indirect way to select for resilience. Resilience has been directly related to welfare. A good body condition and efficient body fat mobilization have been associated with an optimal level of animal welfare. Two rabbit lines have been divergently selected for litter size residual variability. The low line selected for decreasing litter size variance more efficiently managed the body fat from mating to weaning in the second productive cycle in females compared to the high line, which could be related to the lower culling rate reported previously in the low line. Therefore, body condition can be used as a useful biomarker of resilience. A divergent selection experiment for residual variance of litter size at birth was carried out in rabbits during twelve generations. Residual variance of litter size was estimated as the within-doe variance of litter size after pre-correction for year and season as well as parity and lactation status effects. The aim of this work was to study the correlated response to selection for litter size residual variability in body condition from mating to weaning. Body condition is related directly to an animal's fat deposits. Perirenal fat is the main fat deposit in rabbits. Individual body weight (IBW) and perirenal fat thickness (PFT) were used to measure body condition at second mating, delivery, 10 days after delivery, and weaning. Litter size of the first three parities was analyzed. Both lines decreased body condition between mating to delivery; however, the decrease in body condition at delivery was lower in the low line, despite this line having higher litter size at birth (+0.54 kits, p = 0.93). The increment of body condition between delivery and early lactation was slightly higher in the low line. On the other hand, body condition affected success of females' receptivity and fertility at the third mating, e.g., receptive females showed a higher IBW and PFT than unreceptive ones (+129 g and +0.28 mm, respectively), and fertile females had a higher IBW and PFT than unfertile ones (+82 g and +0.28 mm, respectively). In conclusion, the does selected for reducing litter size variability showed a better deal with situations of high-energy demand, such as delivery and lactation, than those selected for increasing litter size variability, which would agree with the better health and welfare condition in the low line.This research was supported by Project AGL2017-86083-C2-2-P, funding by Ministerio de Ciencia e Innovacion (MIC)-Agencia Estatal de Investigacion (AEI) and el Fondo Europeo de Desarrollo Regional (FEDER).Agea, I.; García, MDLL.; Blasco Mateu, A.; Massanyi, P.; Capcarová, M.; Argente, M. (2020). 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