High temperature impairs rabbit viability, feed consumption, growth and fecundity: examination of endocrine mechanisms.

The aim of the present study was to examine the effect of high ambient temperature on rabbit feed consumption, growth, viability, and fecundity, as well as the morphology and endocrine function of gonadal and adrenal cells. Adult does and their offspring were kept at either a comfortable (20°C; control) or high (36°C) temperature throughout pregnancy and up until weaning of pups. Doe mortality and fecundity, and plasma concentrations of hormones were evaluated. In addition, granulosa cells were cultured with and without FSH to assess progesterone production. In the offspring, we assessed mortality, total feed consumption, feed efficiency, growth, plasma hormone concentrations, as well as the microstructure in ovarian granulosa cells, testicular Leydig cells, and adrenocortical cells. We observed greater mortality of both adult animals and offspring at the higher ambient temperature compared with the control. The higher ambient temperature suppressed feed consumption, feed efficiency, and growth of pups. Adult and young females exposed to a high temperature had lower circulating concentrations of progesterone, but not of estradiol, compared with controls. Young males exposed to a high ambient temperature had greater circulating concentrations of testosterone, but not progesterone, compared with controls. High ambient temperature reduced circulating IGF-I concentrations in all the animals. Corticosterone level was increased in plasma of young but not of adult animals. Granulosa cells isolated from the ovaries of does subjected to high temperatures released less progesterone, and they had poorer response to the stimulatory action of FSH than the cells from control does. High temperatures induced fragmentation of nucleoli in ovarian granulosa cells, but they did not alter the state of other organelles in ovarian, testicular, or adrenocortical cells. A negative influence of high temperature on rabbit feed consumption, growth, viability, and fecundity was observed. Taken together, these changes could be due to a decrease in IGF-I and/or progesterone secretion, destruction of ovarian cell nucleoli, and/or impaired ovarian cell response to FSH.

Multilevel D-loop PCR identification of hunting game.

The 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 25 µl at conditions 95 °C for 2 min, 94 °C for 30 s, 60 °C for 30 s, 72 °C for 30 s, 35 cycles, with last extension at 72 °C for 10 min. D-loop mtDNA amplicons of the game animals are characterized with specific PCR product sizes depending on species: red deer = 163 bp and 140 bp, fallow deer = 280 bp and 138 bp, roe deer = 303 bp, 280 bp, 160 bp and 138 bp, mouflon = 299 bp and 178 bp, wild boar = 137 bp and 229 bp.

Changes of femoral bone tissue microstructure in transgenic rabbits.

Bone tissue microstructure of femur was investigated in transgenic New Zealand White rabbits with human factor VIII gene. Altogether 42 bones (24 from transgenic rabbits and 18 from non-transgenic ones) were analysed. Specimens were prepared using standard histological equipment, producing thin sections of approximately 80-100 microm. For histomorphometrical analysis areas, perimeters, minimum and maximum diameters of osteons' vascular canals and of osteons were measured. We found out that the basic structural pattern of femoral bone tissue was primary vascular longitudinal in both groups of rabbits. However, a new type of the bone tissue--fibrolamellar--was identified only in the transgenic rabbits. The measured variables of the osteons' vascular canals were higher in transgenic individuals in comparison with the nontransgenic ones (except for maximal diameter) and the differences were statistically significant (P < 0.05; P < 0.01). We suppose that the observed differences could be associated with transgenesis. In an effort to explain these differences we compared the cytogenetic profile of bone marrow cells between transgenic and non-transgenic rabbits. A significantly higher rate of aneuploidy was observed in c-metaphase spreads of transgenic individuals as compared to non-transgenic ones (P < 0.001). Despite the fact that no hFVIII mRNA expression was found in the femur of transgenic rabbits, we discussed an association of transgene integration into the genome and microstructural changes in the bone. In any case, the results indicate that transgenesis can also produce changes in other tissues than in the target ones.

Aneuploidy in the transgenic rabbit.

The aim of this study was to determine whether there are differences in the karyotypes between transgenic and non-transgenic or control rabbits. New Zealand White transgenic rabbits (F1 generation) were obtained after breeding of transgenic founder rabbits that were derived from single--SM--or double microinjection--DM--with a WAP-hFVIII transgene. C-metaphase plates were obtained from short-time culture of peripheral blood lymphocytes synchronized by the addition of colcemide. A significantly higher rate of aneuploidy was observed in c-metaphase spreads of transgenic (56-66%) rabbits, as compared to non-transgenic ones (28-38%) (P < 0.05; P < 0.01). The patterns of chromosome banding were identical in both groups of rabbits. No structural aberrations were revealed in either group. These findings demonstrate that transgenic rabbits have a higher frequency of numerical chromosomal aberrations in their peripheral blood lymphocytes than normal rabbits, but without apparent deleterious effects on health or reproduction.

A technique to examine cock sperm chromosomes using zona-free hamster eggs.

For the first time the visualization of cock sperm chromosomes using zona-free hamster eggs was described. Different variants of egg treatment were tested: varying the duration of incubation of eggs with spermatozoa from 5-6 to 20 h and the cytostatic treatment (colchicine) from 35 min to 9 h, varying the concentration of colchicine from 0.4-400 micrograms/ml and the hypotonic treatment with 0.1% and/or 1.0% sodium citrate or with distilled water from 2-5 to 12 min. In all variants sperm penetration was found, with changes in the head of sperm and the formation of pronucleus but there was no breakdown of the pronuclear envelopes. Only experiments, in which eggs were incubated without colchicine, were successful.

[The effect of magnesium sulfate used for killing rabbits on aldolase activity in organs]. / Vplyv síranu horecnatého pri usmrtení králikov na aktivitu aldolázy v ich orgánoch.

The effect of the system of mating and the way of killing on the aldolase activity in the tissue of rabbits was the task of this research work. The research work was realized in 201 New Zealand white rabbits aged 140 days, mated outbred and inbred. Two methods of killing were used: classical method, which was based on the stroke and break the spinal cord; and killing with the help of treating with an i/m infection of 10 per cent of magnesium sulphate (2 ml per 1 kg of weight). In blood serum and in liver homogenates, kidneys and dorsal muscle the aldolase activity was determined by the method of Bruns. No significant differences in the aldolase activity between outbred and inbred rabbits were found. Statistically high significant differences, conditioned by killing method, were stated in activity of enzyme.