R. Cardinali

Effects of flaxseed dietary supplementation on sperm quality and on lipid composition of sperm subfractions and prostatic granules in rabbit

Lipids are the main structural/functional components of the sperm, and their composition may undergo a series of modifications in relation to either physiologic events (capacitation and acrosome reaction) and/or diet. The goals of the current study were (1) to investigate whether a flaxseed (FS) dietary supplementation could affect the lipid and fatty acid profile of sperm subfractions and of prostatic granules (PGs) and (2) to evaluate the effects of dietary FS on rabbit buck semen quality. Accordingly, 20 adult New Zealand White rabbits were fed ad libitum a control diet (CO) or a diet supplemented with 5% extruded FS. Integration of diet with FS, as a consequence of the linolenic acid (C18:3n-3; LNA; 56%), increased the dietary n-3/n-6 ratio and resulted in a substantial rearrangement of sperm fatty acid composition at the subcellular level, mainly of polyunsaturated fatty acid (PUFA)n-3 (8.3% vs. 14.3%, P<0.05). The lipid and fatty acid profiles of sperm tail membrane were the most affected, undergoing the following significant changes: (1) a reduction by half of linoleic acid (C18:2n-6; LA) and docosapentaenoic acid (22:5n-6; DPA), and a reduction of cholesterol (-70%); (2) a concomitant increase of LNA (+65%), docosahexaenoic acid (22:6n-3; DHA; +83%), and of oleic acid (C18:1n-9, +61%). As a consequence, the sperm of FS-fed rabbits had a twice higher n-3/n-6 ratio and phospholipid/cholesterol ratio compared with the control sperm. These changes might have been on the basis of the higher responsiveness to hypo-osmotic solution and, hence, the higher sperm track speed observed for the FS group. Also, the membrane integrity and viability of the LNA-enriched sperm were both improved. On the other hand, the presence of lignans in FS might have accounted for the reduction of sperm cholesterol in the semen of FS-treated rabbits. The responsiveness of sperm to acrosome reaction was not affected by the dietary treatment probably due to supranutritional level of vitamin E and to the higher number of PGs, which are known to play a key role in sperm capacitation. In conclusion, our data showed for the first time that the integration of FS into the rabbit diet may improve sperm quality by modifying the sperm lipid composition and that the sperm subfractions and the PGs respond differently to the FS-induced lipid manipulation.

In vitro toxic effects of metal compounds on kinetic traits and ultrastructure of rabbit spermatozoa.

Metal compounds have been associated with male reproductive toxicity in vivo. The aim of the present study was to investigate the in vitro effects of 20 metal compounds using rabbit ejaculated spermatozoa as a study model for spermiotoxicity. Five of the metals tested (arsenic, cadmium, chromium, mercury and vanadium) reduced sperm motility and curvilinear velocity. Ultrastructural analyses revealed three types of damage to sperm head membranes in relation to the metal used: acrosome breakage with formation of various sized microvesicles (arsenic, cadmium, mercury and platinum); a large round hole (arsenic, cadmium and chromium), and numerous folds in the acrosome membrane (vanadium). The vanadium compound, followed by chromium and mercury compounds, determined a higher number of damaged spermatozoa. In conclusion, all the studied metal compounds, at levels higher than 1microM, may reduce sperm kinetic characteristics and probably fertilizing capacity by triggering specific morphological damages to the head and/or by inhibiting motility.

Effect of dietary supplementation with olive pomaces on the performance and meat quality of growing rabbits

The aim was to investigate the effects of three types (A, B and C) of stoned and dehydrated olive pomaces (OPs), differing in olive cultivar, on productive performance and meat quality of growing rabbits. The inclusion of OPs (5%) negatively affected the performance of rabbits as it reduced the feed intake, growth rate, carcass weight and dressing out percentage (P<0.05). Compared with the control, the meat of OP rabbits had a greater amount of monounsaturated and a lower amount of polyunsaturated fatty acids (P<0.05), independent of the type of OP used. Oxidative processes in the meat of OPA and OPB were higher (P<0.05), whereas OPC showed the same levels as the control group. This was due to the higher total polyphenol concentration and to the concomitant lower peroxide value of OPC. These results recommend the use of OP in rabbit diet with caution, taking into account the quality of the by-product in terms of oxidative status.

The main factors affecting the reproductive performance of rabbit does: a review.

This paper reviews the main factors affecting the reproductive performance of rabbit does. In the last 15 years the profitability of rabbit farms has increased mainly due to improvements in management and genetic selection but several problems related to animal welfare have also occurred. The replacement and the mortality rates of female per year are very high and the replaced does often show poor body condition and poor health status. The effect of kindling order, litter size, genetic strain, weaning age and reproductive rhythm on the reproductive performance and welfare of females and mechanisms implicated in these effects are discussed. The rabbit doe of modern strains produces a lot of milk with high energetic value, which leads to a mobilization of body fat resulting in a negative energy balance. In the current reproductive rhythms, there is an extensive overlap between lactation and gestation. The resulting energetic and hormonal antagonism reduces the fertility rate and lifespan of the doe. Strategies to optimize these parameters are discussed. An approach that combines various strategies seems to be required to meet these objectives. Since the factors involved in this system are predetermined (genetic strain, environment) the most powerful way to improve doe welfare is to choose a reproductive rhythm more adapted to the physiology of the does.

Short- and long-term effects of lipopolysaccharide-induced inflammation on rabbit sperm quality

Infections and resulting inflammation are widely known to cause transient or permanent male infertility. The objectives of this study were (1) to provide a suitable animal model of a sub-acute inflammatory state by intraperitoneally inoculating bacterial lipopolysaccharides (LPS) and (2) to define the short- and long-term effects of this state on the sperm quality of rabbit bucks. Two series of experiments were performed to accomplish these objectives. In experiment 1, 15 healthy New Zealand White rabbit bucks were divided into five homogeneous groups, receiving 25, 50, 100 and 150 microg/kg body weight (b.w.) of E. coli LPS dissolved in 2ml of sterile saline or only saline (control), respectively. White blood cells (WBC), rectal temperature, feed intake and mating ability were observed for 3 consecutive days following inoculation. Inoculation of 50 microg/kg b.w. produces a reversible inflammation-like state that lasts for about 3 days, with minimal distress to the animals, and therefore it was used in our experiment. The major symptoms were fever and anorexia. Changes in WBC count and a moderate reduction in reproductive activity also occurred. In experiment 2, two groups of five rabbit bucks each were treated with 50 microg/kg b.w. E. coli LPS diluted in 2ml of saline or only saline (controls), respectively. Semen samples were collected weekly up to 56 days after inoculation and the changes in semen characteristics were examined. During the first 3 days, semen volume and concentration decreased in both experimental groups, probably due to the high collection frequency. Sperm membrane integrity and the number of necrotic sperm were seriously affected 30 days after the LPS challenge, reaching a maximum at the end of the spermatogenic cycle (56 days). These results suggest that a sub-acute inflammation may cause infertility by compromising sperm membrane integrity which decreases a month after LPS-treatment. In addition, the rabbit could be a useful LPS animal model for further study of the effects of inflammation and the underlying mechanisms on sperm characteristics.

Effect of genotype and rearing system on the native immunity and oxidative status of growing rabbits

Abstract To investigate the effects of genotype and rearing system on innate immunity and oxidative status, 40 weaned commercial hybrid (H) rabbits and 40 rabbits of a local population (LP) were assigned to two rearing systems: cage (17 rabbits/m2) and pen (1x2 m, 10 rabbits/m2). Rearing H rabbits in pen resulted in a higher lysozyme concentration (P<.05), an increased bactericidal activity (P<0.05) and a lower complement haemolytic activity (P<0.05) with respect to those reared in cage. Opposite results were obtained for LP rabbits. In addition, genotype H was more susceptibility to inflammation than LP in pen, whereas genotype LP suffered more in cage. A higher lipid (P<0.05) and protein (P<0.05) oxidation was found in Longissimus dorsi and Biceps femoris of rabbits H in pen compared to those in cage. An opposite trend was observed for LP rabbits. In cage, LP rabbits were more susceptible to oxidative stress than H rabbits. Taken together, our results show that immune and oxidative resistance ascribed to a specific genotype are important factors to be considered when a specific rearing system should be adopted. Considering this, rabbit LP seem to better adapt to larger spaces during growing.

In vitro antioxidant activity of the prostatic secretory granules in rabbit semen after exposure to organic peroxides

BackgroundThe prostate gland of rabbits produces numerous granules, which are specifically implicated in the inhibition of sperm capacitation during the first hours after mating. These granules are rich in vitamin E, but their role in the antioxidant protection of rabbit sperm has not been studied.Aim of studyThe objectives of this study were to investigate whether the prostatic secretory granules (PSGs) could prevent sperm induced-lipid peroxidation and to verify a potential involvement of tocopherols from the PSGs to the sperm.MethodsWashed sperm samples taken from eighteen White New Zealand rabbits were either incubated with tert-butyl-hydroperoxide (t-BHP, an oxidative stressor) or with buffered Tyrodes medium for 1 hour. The same number of sperm samples that contained PSGs were subjected to the previously mentioned treatments and thiobarbituric acid reactive substances (TBARS), vitamin E compounds and the acrosome status were assessed.ResultsThe incubation of the sperm with t-BHP resulted in a noticeable production of TBARS (0.38 vs. 0.22 nmol/10(7) cells) and an associated decrease of alpha-tocopherol (alpha-T, 72.3 vs. 103.2 nmol/10(8) cells) with respect to the sperm samples containing PSGs. The sperm incubated with the PSGs had a higher amount of alpha-T compared to the control (292.2 vs. 251.0 nmol/10(8) cells). The acrosome status was not affected by the occurrence of the organic peroxide in the medium and the amount of capacitated sperm was lower when the PSGs were also present.ConclusionsOverall, these results suggest that the PSGs may represent a source of protection for rabbit sperm against in vitro oxidative stress by supplying the sperm with endogenous alpha-T. This mechanism could be in part involved in the inhibition of sperm capacitation by the granules.

Food restriction during pregnancy in rabbits: Effects on hormones and metabolites involved in energy homeostasis and metabolic programming

This study examined the effects of food restriction during rabbit pregnancy on hormones and metabolites involved in energy homeostasis and metabolic programming. Pregnant does were assigned to four groups: the control group was fed a standard ration while the others received a restricted amount of food (30% restriction) during early (0-9 days), mid (9-18 days), and late (19-28 days) pregnancy. The pregnancy induced a coordinated range of adaptations to fulfil energy requirements of both mother and foetus, such as hyperleptinaemia and hyperinsulinaemia, reduced insulin sensitivity, increased cortisol and non-esterified fatty acid. Food restriction altered leptin, insulin, T3, non-esterified fatty acids and glucose concentrations depending on the gestational phase in which it was applied. Collectively, present data confirm that the endocrinology of pregnancy and the adaptive responses to energy deficit make the rabbit an ideal model for studying nutritional-related disorders and foetal programming of metabolic disease.

Effect of a Bacterial Lipopolysaccharide Treatment on Rabbit Testis and Ejaculated Sperm

In a previous study, we reported the short- and long-term effects of bacterial lipopolysaccharide (LPS)-induced inflammation on rabbit sperm quality. This study was aimed at exploring the spermatogenesis of the rabbit model focussing on the possible damages occurring to the testis and ejaculated sperm. Twenty New Zealand White rabbit bucks were divided into two groups. One group was inoculated intra-peritoneally with LPS, the other group, considered as control, was treated under the same conditions with saline only. Semen samples were collected before LPS injection, the 7th, 14th, 21st, 30th, 45th, 60th and 90th day after LPS treatment. Semen parameters were evaluated following international guidelines. The kinetic characteristics of ejaculated sperm were analysed using a computer-assisted sperm analyzer and the ultrastructural characteristics were explored by transmission electron microscopy (TEM). On the 7th, 14th and 30th day, testis from treated rabbits and controls were obtained. Testis samples were analysed by light microscopy and TEM. The induced LPS lesions in the testis became evident the 7th day after treatment, with a decrease in germinal cells and with an increase in structurally altered Sertoli cells; normal spermatogenesis was restored on the 30th day. The testicular damages observed on day 7 were probably responsible for the reduction in sperm concentration and motility and the ultrastructural alterations that were detected in the ejaculated sperm on the 14th through the 30th days after treatment. In conclusion, rabbit buck treated with LPS could be a useful model for studying the effect of an induced systemic inflammation on spermatogenesis.

Effect of chocolate and Propolfenol on rabbit spermatogenesis and sperm quality following bacterial lipopolysaccharide treatment.

Abstract The aims of the study were to evaluate the effects of chocolate and propolis-enriched diets on rabbit spermatogenesis, sperm motility, and ultrastructure following bacterial lipopolysaccharide (LPS) treatment. Thirty-two New Zealand White rabbits were divided into four groups. The LPS-Propolfenol® group received propolis (500 mg/kg/day) in their diet for 15 days, while the LPS-chocolate group was fed 70% cacao chocolate (1 g/1 kg/day) for the same period. Following the diet treatments, rabbits in the LPS-Propolfenol® and LPS-chocolate groups, and an LPS group received a single intraperitoneal dose of 50μg/kg LPS, and the control group received only saline. Kinematic sperm traits were evaluated with a computer assisted sperm analyzer (CASA) system, and ultrastructural characteristics were examined by transmission electron microscopy (TEM). Testicular and epididymal tissues were observed by light microscopy and TEM and multiplex real time reverse transcriptase-polymerase chain reaction (RT-PCR) assay was used to detect and quantify toll-like receptor-4 (TLR-4) gene expression. The values of the analyzed semen parameters of rabbits treated with LPS-Propolfenol® and LPS-chocolate did not show any variations compared with the control group, but they were lower in rabbits treated only with LPS. Alterations observed in the testicular tissue of LPS treated-rabbits were not detected in specimens from the LPS-chocolate and LPS-Propolfenol® groups, which showed normal spermatogenesis. The TLR-4 mRNA expression was similar in controls, in LPS treated, and in LPS-chocolate groups, but it was significantly (p < 0.01) decreased in LPS-Propolfenol® rabbits. In conclusion, a chocolate and propolis-enriched diet showed a protective effect on the spermatogenetic process of buck rabbits following LPS treatment.