Multilevel D-loop PCR identification of hunting game

AbstractThe control region of mtDNA (D-loop) was used for hair samples of the five hunting game species identification: red deer (Cervus elaphus), roe deer (Capreolus capreolus), fallow deer (Dama dama), mouflon (Ovis aries musimon), and wild boar (Sus scrofa). For D-loop multilevel PCR detection scheme was applied in six primers (CE CVZV 1=5′-GATCACGAGCTTGATCACCA-3′; CE CVZV 2=5′-AGGAGTGGGCGATTTTAGGT-3′; DD CVZV 3=5′-CGCGTGAAACCAACAACCCGC-3′; DD CVZV 4=5′-CCGGGTCGGGGCCTTAGACG-3′; SSW CVZV 5=5′-ACACGTGCGTACACGCGCATA-3′; SSW CVZV 6=5′-GGTGCCTGCT T TCGTAGCACG-3′) designed to identify unknown biological samples of the hunting game animals. The PCR reaction volume was 25μl at conditions 95°C for 2min, 94°C for 30s, 60°C for 30s, 72°C for 30s, 35cycles, with last extension at 72°C for 10min. D-loop mtDNA amplicons of the game animals are characterized with specific PCR product sizes depending on species: red deer=163bp and 140bp, fallow deer=280bp and 138bp, roe deer=303bp, 280bp, 160bp and 138bp, mouflon=299bp and 178bp, wild boar=137bp and 229bp

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

Decrease in C-reactive protein levels in rabbits after vaccination with a live attenuated myxoma virus vaccine

The aim of this study was to evaluate the acute phase reaction and immune response of rabbits triggered by vaccination with a live attenuated myxoma virus (MXT) vaccine. Thirteen adult and 11 juvenile New Zealand white rabbit-based crossbreed rabbits, were used. Samples of rabbit peripheral blood were collected from vena auricularis centralis into heparinised tubes before vaccination and 48 h after vaccination. All animals were vaccinated by subcutaneous injection (0.5 ml) with a MXT vaccine. The blood plasma C-reactive protein level was measured by an ELISA kit using a double-antibody sandwich. For phenotyping of lymphocytes the fresh cells were stained with the following anti-rabbit monoclonal antibodies: anti-IgM, anti-CD4, anti-CD8 and anti-pan T2. Our results show that the use of attenuated myxoma virus vaccine significantly decreases the level of C-reactive protein in blood plasma of adult rabbits by 38.14% (P < 0.05) and of juvenile rabbits by 37.63% (P < 0.001), within 48 h. The rabbit C-reactive protein after MXT vaccination is a negative acute phase protein. In the group of adult rabbits the immune response to MXT vaccination was accompanied by a non-significant decrease in CD4+, pT2+, IgM+ subsets. On the other hand the values of CD8+, CD4+CD8+ and CD4+/CD8+ were non-significantly higher after MXT vaccination

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

Effective generation of genetically modified rabbits by sperm mediated gene transfer

[EN] Many reports in the past decade showing successful sperm mediated gene transfer (SMGT) offoreign DNA into both non-mammalian and mammalian animals. Fresh collected, extender washed rabbit spermcells were co-cultured with linear 14.3 kb gene construct and used for AI. Transgene integration in rabbitgenome was verified by PCR analysis. The PCR results show that transgenic founder rabbits transmittedtransgene to F1 generation. Our preliminary results suggest that SMGT could be an effective method forproduction of genetically modified rabbits.We would like to thank Mr. Jan Pecho and Dipl. Ing. Ivan Chlebec for taking care of the animals and for providing technical assistance. This work was financially supported by Ministry of Agriculture Slovak Republic and Slovak Research and Development Agency under the contract No. APVV-27-005505.Vasicek, D.; Vasickova, K.; Parkanyi, V.; Rafay, J.; Ondruska, L. (2007). Effective generation of genetically modified rabbits by sperm mediated gene transfer. World Rabbit Science. 15(3). https://doi.org/10.4995/wrs.2007.593SWORD15

Effect of dietary Lippia citriodora extract on reproductive and productive performance and plasma biochemical parameters in rabbit does

Two doses of natural extract from Lippia citriodora (titrated in verbascoside) were assessed in New Zealand White rabbit does evaluating selected reproductive, productive and plasma biochemical parameters. After 1 week of adaptation period, the trial on 45 rabbit does for three consecutive reproductive cycles was conducted; does were divided into three groups of 15 animals each, homogenous by age (1 year ± 2 weeks), bodyweight (4.77 ± 0.21 kg) and parity (2 ± 1). A control group (CON) did not receive the dietary supplement in the feed and the other two groups received 1 g of natural extract supplement in the feed (5 mg verbascoside/kg feed; LNE) and 2 g of natural extract (10 mg verbascoside/kg feed; HNE). The use of a dietary NE supplement improved kit bodyweight at weaning (934 vs 1104 g; P < 0.001), and average daily weight gain from birth to weaning (24.7 vs 29.7 g/day; P < 0.001), with no NE dose effect. In the LNE and HNE groups serum triglycerides, total cholesterol, low density lipoprotein cholesterol, bilirubin, and activities of alkaline phosphatase, alanine aminotransferase (P < 0.05) and aspartate aminotransferase (P < 0.01) decreased and high density lipoprotein cholesterol (P < 0.01) increased, according to the cycle effect. The dietary supplement also improved blood oxidative status markers in the experimental groups due to an increase in the concentrations of plasma vitamin A and E (P < 0.01) and a decrease in plasma reactive oxygen metabolites and thiobarbituric acid reactive substances values (P < 0.01). In conclusion, the dietary Lippia NE supplement improved selected productive and reproductive parameters and the animal welfare of does, expressed by a general improvement of blood profile, with no effect of the dose