Academic literature on the topic 'Rabbits Spermatozoa'

SummaryIntracytoplasmic sperm injection (ICSI) is an assisted fertilization technique and has been widely applied in human medicine to overcome some obstacles of infertility. However, this technology has not yet been used as a mainstream technique for animal production, including the rabbit, due to its limited success. The aim of this study was to improve ICSI techniques and establish an efficient ICSI method for rabbits. Spermatozoa used for ICSI were collected from mature New Zealand white male rabbits. They were washed two to three times with HEPES-buffered TCM199 containing 10% FBS and then mixed with 10% polyvinylpyrrolidone (PVP) prior to microinjection. Oocytes were harvested from superovulated donor rabbits after 14–15 h hCG treatment and were fertilized by microinjection of a single living spermatozoon into the ooplasm of each oocyte without additional activation treatment. A total of 317 injected oocytes resulted in the high survival rate of 86.1%. Among the surviving oocytes, 273 were placed into culture dishes for in vitro development. The fertilization, cleavage and blastocyst rates were 59.0%, 88.2% and 45.3% respectively. Furthermore, ICSI embryos were produced with spermatozoa from an infertile male rabbit, and 21 early-stage embryos (2-cell and 4-cell) were surgically transferred into the oviducts of two adult female rabbits. On day 31 after transfer, one out of the two recipients gave birth to two normal and healthy young rabbits. These results demonstrate that rabbit oocytes can be successfully activated and fertilized by the new ICSI protocol. Spermatozoa derived from infertile rabbits can successful fertilize oocytes and produce offspring by the simple ICSI technique.

The purpose of this study was to determine the concentration of spermatozoa in New Zealand rabbits after giving of various types of beluntas leaf tannin extract. This type of research is experimental research. The research design is a one-group post-test design. A sample of 16 rabbits. The sampling technique used is purposive random sampling. The research was conducted at the Chemical Engineering Laboratory of the State Polytechnic of Malang, the Laboratory of Kesimma Medica Malang, the Integrated Laboratory, and the Biology Laboratory of the University of Muhammadiyah Malang. The study was carried out from August to September 2019. The study design used a completely randomized design (CRD) with 4 treatments, namely hydrolyzed, condensed, and pure tannin extract of beluntas leaves and the control group was given aquades orally with a dose of 3 ml/kgbb with 4 replications. repetition times. The procedure of this research 1) Preparation of extract of beluntas leaf tannins, 2) Phytochemical test of extract preparations, 3) Treatment of rabbits as experimental animals, 4) Surgery and collection of rabbit spermatozoa, 5) Observation of spermatozoa using a microscope, 6) Calculating the concentration of spermatozoa. The method of data collection was by calculating the concentration of spermatozoa in each treatment using a hemocytometer and a Neubauer counting chamber using a binocular microscope. Data analysis with one-way Anova test and Duncan 5%. The results showed that the dosage of 3ml/kgbb of various types of beluntas leaf tannin extract had an effect on the spermatozoa concentration of New Zealand rabbits (sig<0.05). The findings of this study are that the best treatment that can reduce the number of spermatozoa in New Zealand rabbits is the condensed tannin extract of Beluntas leaves.

SummaryThe aim of our study was to compare the viability of sperm cells from transgenic (mWAP-hFVIII gene) or non-transgenic (normal) rabbit males as assessed by viability (SYBR-14/PI) and apoptosis (annexin V) tests. These results were evaluated using female conception rates following insemination with the respective sperm samples. No significant differences were found in concentration and motility between transgenic and non-transgenic spermatozoa. Spermatozoa from both transgenic (63.05 ± 20.05%) or non-transgenic (65.75 ± 22.15%) males, stained with SYBR-14 (green), were found to be morphologically normal. In both groups, the highest proportion of annexin V-positive sperm staining was found in the post-acrosomal part of the sperm head (8.66 and 27.53%). The percentage of sperm that stained with SYBR-14/PI or with annexin V/DAPI was correlated with liveborn in transgenic rabbits (R2 = 0.6118 and R2 = 0.2187, respectively) or non-transgenic rabbits (R2 = 0.671 and R2 = 0.3579, respectively). These data indicate that there was no difference in the viability of rabbit transgenic and non-transgenic spermatozoa when determined by both fluorescence assays.

Abstract The study evaluated the effect of Vernonia amygdalina leaf meal on semen indices, serum testosterone and sperm reserve of male rabbits. Forty rabbit bucks were randomly assigned into four groups and fed the experimental diets containing Vernonia amygdalina (VLM) at 0, 5, 10 and 15 % levels for 84days. Reproductive indices were evaluated using standard procedures. Data obtained were subjected to analysis of variance at p<0.05. All semen in rabbits fed 0, 5 and 10 % VLM had milky colour while 14.8% light green and 85.2% milky semen colour was observed in bucks fed 15%VLM. Libido score reduced in rabbits fed VLM diets. Bucks fed 15%VLM had significantly higher semen volume (0.47ml). VLM had no significant effect on spermatozoa mass motility, progressive motility and sperm concentration. Live sperm cells significantly increased in bucks fed 5 and 10% VLM diets. VLM had significant (p<0.05) effect on spermatozoa morphology. Vernonia amygdalina leaf meal had no significant (p<0.05) impact on testosterone, testicular and epididymal indices. In conclusion, up to 10%VLM can be adopted as feed ingredient for male rabbit breeder stock without deleterious effect on reproductive indices.

Some researchers have inferred that folic acid is necessary for reproduction and could enhance blood formation. Thus, a Completely Randomized Design Experiment (CRD) was conducted to evaluate the impact of oral administration of folic acid on the semen and haematological characteristics of New Zealand White rabbit bucks. The treatments designated treatment 1 (T1), treatment 2 (T2) and treatment 3 (T3) having 12 rabbits each were replicated 3 times with 4 rabbits per replicate. The ages of the 36 pre-pubertal rabbit bucks were between 2 to 3 months, and weighed approximately 2.56 kg. Three experimental diets were formulated to meet the nutrient requirements of rabbit bucks. Each rabbit buck on T1 were orally administered folic acid at 0.0 mg, T2 2.5mg folic acid and T3 5.0 mg folic, respectively. Data were collected for semen characteristics and haematology from the rabbit bucks. Data collected on different parameters were subjected to analysis of variance (ANOVA). Results showed that significant increases (p<0.05) were observed on libido, semen pH, spermatozoa progressive motility (67.40-80.20%), spermatozoa live proportion (83.01-94.12%), sperm concentration (112.24-133.80 x106/ml), total number of sperm per ejaculate (50.65-67.66 x106/ml), total viable sperm (291.58-496.69 x109/ml), normal sperm proportion (85.16-91.64%). Also, significant reductions (p<0.05) were observed on the percentage head abnormality of the spermatozoa (3.74-3.18), total abnormality (2.13-0.93), mid-piece abnormality (2.35-0.79), cytoplasmic abnormality (7.17-2.89), and total abnormality (14.84-8.35); while the haematological parameters such as haemoglobin (13.53-14.20g/dl), packed cell volume (33.00-34.96%), white blood cell (6.81-7.80 x103mm3) and the differential white blood cells improved significantly (p<0.05) following the oral administration of folic acids to the rabbit bucks. Thus, the oral administrations of folic acid at 5.0 mg per rabbit buck most significantly improved the semen characteristics, enhanced the overall spermatozoa morphology, reduced sperm cells abnormalities and also improved some haematological parameters of the rabbit bucks.

Transgenic animal mammary gland bioreactors are used to produce recombinant proteins. However, it is difficult to validate whether these transgenic domestic animals are able to express the recombinant protein efficiently in their mammary glands before the birth of transgenic offspring. In the present study, a simple and efficient method was established to evaluate the functionality of animal mammary gland tissue-expressed cassettes. The gene transfer vector pGBC2LF was constructed, and the expression of human lactoferrin (LF) gene was controlled by the goat β-casein gene 5′ flanking sequence. To obtain the most efficient transfection, the influence of DNA concentration, dimethylsulfoxide (DMSO) concentration, and the ratio of linear-to-circular DNA required for associating DNA with spermatozoa were evaluated. Transfection of exogenous DNA into rabbit spermatozoa was found to be efficient using 30 μg mL–1 DNA, DMSO at a final concentration of 3%, and a 3 : 1 ratio of linear-to-circular DNA, with 29 of 85 (34.1%) in vitro-fertilised embryos being transgenic. Using DMSO–sperm-mediated gene transfer (DMSO-SMGT), 89 rabbit offspring were produced, with 46 of these (57.1%) being transgenic. As mammary gland bioreactor models, 17 of 21 (81%) transgenic female rabbits could express human LF protein in their glands. During lactation of the transgenic rabbits, the highest level of human LF protein expressed was 153 ± 31 μg mL–1, and the mean expression level in all of the transgenic rabbits was 103 ± 20 μg mL–1 in the third week, declining gradually after this time. Our results demonstrate that transgenic rabbits produced by DMSO–SMGT were able to express human LF protein in the correct tissue.

Introduction. Using genetic potential of rabbit ovaries and studying patterns of meiotic maturation of gametes in female in vitro is a basis for success in cloning and the creation of transgenic animals, so there is a need for in-depth study of cytomorphological characteristics of oocytes during in vitro embryogenesis. The aim of the research is cytomorphological study of oocytes during embryogenesis, derived from matured rabbits’ ovaries and before the sexual cycle. Materials and methods of research. The ovaries of the rabbits (n = 8) aged 4 months and rabbits aged 11 months, coming into heat (n = 10) were used in the study. All the ovaries derived from females, were at follicular growth phase. Rabbits’ oocyte-cumulus complexes were cultured in vitro during 24 hours in plastic Petri dishes (25 - 30 OCC per ml) in the medium for maturing – 199 with Earle’s solution (Sigma, M 5017), supplemented by 20% heat-inactivated (56ᵒC, 30 min.) homemade estrous cow serum, 0.068 mg/ml kanamycin sulfate, 0.11 mg/ml sodium pyruvate and 0.1 mg/ml glutamine. Granulosa cells derived from the antral follicles without atresia evidence were necessarily added in the culturing medium in amount of 3–5х106 per ml. Received in vitro the ova were fertilized by freshly derived rabbit’s epididymal spermatozoa. Results. As a result of extracting oocytes from all the ovaries (n = 18), 245 OCC were received, including 115 OCC from eight rabbits’ ovaries before the sexual cycle and 130 OCC from ten ovaries of mature rabbits. Analysis of cytomorphological studies found more (P < 0,05) oocyte-cumulus complexes can be received from rabbits’ ovaries during puberty being suited for cultivation than from mature rabbits’ ovaries. After in vitro culturing 85,5 % of OCC (47 of 55) derived from rabbits’ ovaries during puberty and 75.6% of OCC (62 of 82) – from mature rabbits’ ovaries reached MII meiosis. It was revealed the level of in vitro maturation of oocytes was 10 % higher in the group derived from rabbits’ ovaries at puberty, compared with the group derived from mature rabbits’ ovaries. Ripened outside a body the oocytes were fertilized in vitro by freshly derived rabbit’s epididymal spermatozoa. The embryos developed in the both groups, but with a significant difference in the level of division. Cytomorphological research found that the level of 2-4-cell embryos formation in the group of oocytes derived from rabbits at puberty was 68,1% and in group of gametes from mature rabbits – only 46.8 % (P < 0.05). 22.2 % of embryos on average developed to the morula stage in vitro. In terms of embryo development to early morula stage significant difference between the groups wasn’t found. The significant difference between the study groups in the number of zygotes have not passed division block (P < 0.05) was noted. In the group of oocytes derived from mature rabbits’ ovaries the zygotes which have not divided were 23.8 % more. Conclusions. It was found that more oocyte-cumulus complexes (P < 0,05) were received from rabbits’ ovaries during puberty, being suited for in vitro cultivation, than from mature rabbits’ ovaries. And oocytes with degeneration signs, being unsuited for in vitro cultivation, were received more (P < 0.05) from mature rabbits’ ovaries. The level of maturation also was 10 % higher in the group of OCC derived from rabbits’ ovaries at puberty. So, for biotechnology research as oocyte donors more effective is use of rabbits during puberty, which have not yet begun sexual cycle, because significantly more (P < 0,05) fully-fledged oocytes cumulus complexes, being suited to culture outside a body, can be derived from their ovaries which will provide greater percentage of preimplantation embryos.

SummaryThis study assessed the potential efficiency of selected biologically active substances on the motility behavior of rabbit spermatozoa subjected to in vitro induced E. faecalis contamination. Semen samples were collected from 10 male rabbits and the presence of E. faecalis was confirmed using MALDI-TOF Mass Spectrometry. For the in vitro experiments rabbit spermatozoa were resuspended in the presence of 0,3 McF E. faecalis and different concentrations of selected biomolecules (resveratrol - RES, quercetin - QUE, curcumin - CUR, epicatechin - EPI, isoquercitrin - IZO). Sperm motility was assessed using the computer-aided sperm analysis at 0h, 2h, 4h, 6h and 8h. The presence of E. faecalis significantly decreased the motility (P<0.001) when compared to the untreated Control starting at 2h and maintaining this negative impact throughout the entire in vitro culture. Meanwhile, the motility was significantly higher in the experimental samples subjected to E. faecalis together 5 μmol/L RES (P<0.05), 10 μmol/L QUE (P<0.05) as well as 1 μmol/L (P<0.01) and 10 μmol/L CUR (P<0.05) when compared to the Positive Control (4h). No biomolecule was able to maintain the motion comparable to the Negative Control, and none was effective against the rapid decline of sperm motility caused by the presence of E. faecalis during later stages of the in vitro experiment (6h and 8h). We may conclude that RES, QUE and CUR may provide a selective advantage to spermatozoa in the presence of E. faecalis, particularly during short-term rabbit semen handling.

The present study investigated regional modifications of glycosylation status, sperm association and functional significance of N- and O-linked glycoproteins in epididymal luminal fluid of the rhesus monkey (Macaca mulatta). The predominant glycoproteins of the epididymal luminal fluid that increase in the extent of glycosylation or unmasking of exposed epitopes in a region-specific, maturation-dependent manner, included those of 150, 116, 68, 64, 58 (N- and O-linked) and 170 kDa (O-linked). The higher expression of 40 (N-linked), 38 (N- and O-linked) and 60, 56 and 33 kDa (O-linked) glycoproteins in the proximal caput epididymal fluid was followed by alteration or reorganization of 60, 38 and 33 kDa (O-linked) glycoproteins in the distal segments of the epididymis. The association of epididymal fluid glycoproteins with maturing spermatozoa was identified by generating polyclonal antiserum against monkey caudal sperm membrane in female albino rabbits. The antiserum crossreacted strongly with 58 and 33 kDa epididymal fluid glycoproteins of monkeys and also reacted with 116, 68, 58, 56 and 33 kDa glycoproteins from Triton X-100 extracts of human spermatozoa, indicating the presence of antigenically related components in both species. The functional significance of epididymal fluid glycoproteins in sperm functions was investigated by raising antiserum against a heavily glycosylated 58 kDa glycoprotein (MEF1) of caudal epididymal fluid, which crossreacted with the Triton X-100 extracts of epididymal spermatozoa of monkey and ejaculated human spermatozoa on immunoblots. In an in vitro micro-sperm agglutination assay, anti-MEF1 serum agglutinated both rat caudal epididymal spermatozoa and human spermatozoa. MEF1 seemed to be involved in fertilization as demonstrated by inhibition of fertility (100%) in female albino rabbits and rats immunized with this protein. A sperm-agglutinating 58 kDa glycoprotein of rhesus monkey epididymis with functional significance in fertility was identified, thus indicating that it is a potential candidate for contraceptive vaccine development.