Biodiversity is increasingly threatened by intensive agriculture, environmental pollution, extinction of natural habitats and several other factors. Several mammalian species including ungulates have disappeared or are threatened by extinction. However, ungulates play an important role both in the ecosystem and in the economy. In general, species or breeds are considered endangered if their population does not exceed 1,000 individuals. In these cases conservation programmes should be initiated in order to maintain or even increase their number. This review deals with the possibilities and limitations of assisted reproductive technologies (ART) in the conservation of ecologically valuable wild, rare and indigenous ungulates. The methods discussed here are artificial insemination, cryopreservation of semen and embryos, embryo recovery and transfer,in vitroproduction of embryos, as well as micromanipulation techniques including sperm injection, assisted hatching and cloning. Some of these procedures are already being exploited in the breeding of farm ungulates, but more basic information about the reproductive patterns of wild, rare and indigenous animal species is needed before the routine use of ARTs.
The extenders and freezing rates from three different freezing protocols were combined and compared to each other in order to study the post-thawing acrosome integrity and fertility of frozen dog sperm. A commercial bovine TRIS-base extender (TRILADYL) and two self-made canine semen extenders (Norwegian and Dutch) were combined with a conventional bovine and two canine freezing regimes, and acrosome integrity of frozen/thawed spermatozoa was assessed by fluorescein isothiocyanate conjugated peanut agglutinin staining (FITC-PNA). Differences between freezing/thawing protocols were reflected in the proportion of cells with acrosomal damage and not based on motility results. It was concluded that during dog semen cryopreservation extenders had less influence on the post-thawing sperm quality than did the freezing rates. The optimal extender/freezing rate combination (TRILADYL/Norwegian) was used in the clinical practice to evaluate the fertility of frozen sperm administered by intrauterine insemination using a surgical approach. The pregnancy rate was 57% (4/7), but the average litter size was low (2.8). This may have been due to the insufficient sperm numbers contained in an insemination dose and/or to the incorrect timing of artificial insemination (AI). The final conclusion is that the commercial bovine extender is useful for freezing dog semen, and the TRILADYL/Norwegian freezing protocol is recommended as the most advantageous combination for the freezing of canine semen in the clinical practice.
Authors:Ádám Nagy, Tassos Polichronopoulos, András Gáspárdy, László Solti and Sándor Cseh
Motility is one of the most important characteristics associated with the fertilising ability of spermatozoa indicating their viability and structural integrity. Therefore, the examination of motility constitutes an integral part of semen analysis. Computer-assisted semen analysis (CASA) allows an accurate and objective assessment of different sperm motion characteristics with high repeatability. The aim of this study was to evaluate the different kinematic (velocity) parameters of frozen/thawed bull semen and determine if any of them could be correlated with their fertilising capability after insemination based on the achieved pregnancy rate. Ejaculates from 10 bulls were collected and frozen. The kinematic/velocity parameters of spermatozoa were measured by CASA and compared to the pregnancy results of almost 9,000 females artificially inseminated (AI) with frozen semen of any of the 10 tested bulls. The data of the experiments are summarised mainly with a focus on the effects of individual velocities (curvilinear velocity: VCL, straight-line velocity: VSL, average path velocity: VAP) on fertility rather than on the influence of progressive motility as a whole. We conclude that VAP is the most useful semen motility characteristic which has clinical relevance in the prediction of fertility.
Authors:Vera Faigl, Nóra Vass, András Jávor, Margit Kulcsár, László Solti, Georgios Amiridis and Sándor Cseh
Artificial insemination (AI) can undoubtedly be regarded as the oldest and most widely used assisted reproductive technique/technology (ART) applied in livestock production and it is one of the most important ARTs. The three cornerstones of its application are that it is simple, economical and successful. Artificial insemination offers many well-known benefits for producers. Fresh, fresh + diluted + chilled and frozen semen can be used for AI in small ruminants. To ensure its successful use, the AI technique must be selected on the basis of the type of semen planned to be used. This review paper gives a detailed overview of semen processing and its effects on semen quality, as well as of the AI techniques applied in small ruminants and their success rates.
Authors:Vera Faigl, Mónika Keresztes, Alíz Márton, Hedvig Fébel, Margit Kulcsár, Sándor Nagy, Sándor Cseh, László Solti and Gyula Huszenicza
Seasonal differences in the resumption of postpartum ovarian activity, milk production and periparturient metabolic status were investigated in lactating non-suckling dairy Awassi sheep in two consecutive experiments. In Experiment 1, autumn-lambing (AL, n = 27) and spring-lambing (SL, n = 37) ewes were investigated. Ovarian activity was monitored by means of individual progesterone (P4) profiles from day 5 to day 100 post partum. Most of the AL dams (89%) ovulated till day 35 after parturition and became cyclic thereafter. Incidence of persistent corpus luteum (CLP) and short luteal phases (sCL) was frequent (18% and 29%, respectively) among non-conceiving dams. In contrast, only 24% of the SL ewes ovulated before day 35. P4 levels during the luteal phase were lower in cyclic animals, and the cycle was longer in SL than in AL animals. No CLP or sCL was detected in the spring-lambing group, and 61% of SL ewes remained acyclic till the end of the trial. Lactation length was significantly longer in SL dams than in AL ewes (P = 0.008). According to the plasma metabolites (BHB, NEFA) and metabolic hormones (insulin, IGF-I, thyroxine) examined, negative energy balance did not appear in any of the animals. However, seasonal differences were seen in IGF-I and thyroxine levels, which were higher in the SL dams. In Experiment 2, influence of additional lighting was studied in autumn-lambing ewes. The long-day photoperiod (LD, n = 23) group was exposed to artificial light from sunset till midnight (approx. 16 h light/8 h dark) from some weeks before the expected date of delivery in mid-September until the end of December. The control group (n = 25) experienced only natural daylength. The first postpartum ovulation tended to occur later in the LD animals than in the controls (P = 0.047). The lactation of the LD group tended to be longer (P = 0.061). NEFA, BHB, insulin, IGF-I and thyroxine levels did not differ between the groups. Conclusions: (i) The ovarian function of the Awassi population is seasonal under temperate continental climate conditions. (ii) The first postpartum ovulation of non-suckling, autumn-lambing dams may occur very early, even before the completion of uterine involution. (iii) Additional artificial lighting may delay the time of first postpartum ovulation in AL ewes. (iv) Postpartum negative energy balance is unlikely to occur in dairy Awassi ewes even in high-producing intensive systems.
Authors:Vera Faigl, Mónika Keresztes, Margit Kulcsár, Sándor Nagy, Zsuzsanna Keresztes, Georgios Amiridis, László Solti, Gyula Huszenicza and Sándor Cseh
The objective of this study was to evaluate the effect of long-term melatonin treatment applied during the non-breeding season on semen characteristics, endocrine function of testicles and baseline level of insulin-like growth factor-I (IGF-I) in Awassi rams kept in the temperate continental zone of Europe and used as semen donors in an artificial insemination (AI) programme. On 23 February (day 0), slow-release melatonin implants were inserted subcutaneously into rams (n = 8). Control animals (n = 8) received no treatment. In both groups, basic semen parameters (concentration, total motility, fast and slow forward motility, morphology), GnRH-induced testosterone response and basal IGF-I concentration were evaluated on days 0, 47 and 71. No differences were found in concentration of spermatozoa, total motility, and numbers of spermatozoa with fast and slow progressive motility and normal/abnormal morphology between the melatonin-treated and the control group. However, in melatonin-treated animals, basal and GnRH-induced testosterone levels were slightly elevated on day 47 and became significantly higher on day 71 (P < 0.05) as compared to controls. There was no difference in plasma IGF-I levels between the groups. In conclusion, slow-release melatonin applied during the non-breeding season improves testicular testosterone production but does not influence the semen characteristics and the IGF-I level of semen donor Awassi rams used in an AI programme and kept in the temperate continental zone of Europe.
Authors:Philip Klambauer, Zsuzsa Keresztes, Katalin Kanyó, Erika Varga, Rita Kriston, Nóra Vass, András Jávor, János Konc, László Solti and Sándor Cseh
By decreasing the volume of the cryoprotective solution it is possible to increase dramatically the freezing speed and — at the same time — reduce the toxicity and osmotic side effects of cryoprotectants (CPA). The objective of our study was to vitrify Day-3 cleavage stage mouse embryos (n = 229) with the cryoloop technology using a new composition of vitrification media. Embryos were exposed to a 2-step loading of CPA, ethylene glycol (EG) and propylene glycol (PG), before being placed on the surface of a thin filmy layer formed from the vitrification solution in a small nylon loop, then they were rapidly submerged into liquid nitrogen. After warming, the CPA was diluted out from the embryos by a 3-step procedure. Survival of embryos was based on morphological appearance after thawing and continued development to expanded blastocysts upon subsequent 48-hour culture. Embryos of the two control groups were either treated likewise except that they were not vitrified, or cultured
without any treatment. Our data show that a high percentage of embryos survived (92.7%) vitrification in the mixture of EG and PG combined with cryoloop carrier and developed normally (89.1%)
after thawing. To our knowledge this is the first report of the successful vitrification of cleavage stage mouse embryos using VitroLoop vitrification procedure.