Relationship between Milk Yield and Reproductive Parameters on Three Hungarian Dairy Farms.

It is postulated that there is negative correlation between milk yield and reproductive performance. However, some studies definitely doubt this causality. The aim of our study was to investigate the relationship between milk production and fertility on three dairy farms. The production parameter was the milk yield (in kg), and fertility was expressed by the number of inseminations per conception (AI index), as well as by the length of the service period (in days). A total of 13 012 lactations from cows with their first three lactations completed were analysed. The number of inseminations was significantly correlated with the milk yield and with the studied farm (p < 0.0001), but its correlation with the lactation number was not significant (p = 0.9477). A similar relationship was found after evaluating the length of the service period. A multiplicative model showed that a 2000 kg milk increase extended the service period by 9% and increased the AI index by 13%. Thereafter, using quartiles of the cows, the service period of the highest-producing group rose by 41.5 days, and the AI index by almost 1, compared to the lowest quartile. Our results indicate a definitive decline in reproductive indicators parallel to an increase in milk production but did not prove an inevitable correlation.

Correlation between bull fertility and sperm cell velocity parameters generated by computer-assisted semen analysis.

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.

Artificial insemination of small ruminants - a review.

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.

Effect of season and photoperiod on the time of first postpartum ovulation in Awassi ewes.

UNLABELLED: 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.

Testicular function and semen characteristics of Awassi rams treated with melatonin out of the breeding season.

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.

Vitrification of cleavage stage mouse embryos by the cryoloop procedure.

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 in vitro 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%) in vitro after thawing. To our knowledge this is the first report of the successful vitrification of cleavage stage mouse embryos using VitroLoop vitrification procedure.

Comparison of follicular and oocyte development and reproductive hormone secretion during the ovulatory period in Hungarian native breed, Mangalica, and Landrace gilts.

Only a very small amount of physiological data is available about the low fertility (mean litter size is 5.7+/-0.8) of Hungarian native breed, Mangalica (M), sows. The aim of the present paper is to reveal the differences in preovulatory follicle development and intrafollicular oocyte maturation between M and Landrace (L) gilts, with special reference to the peri- and postovulatory secretion and peripheral concentrations of estradiol-17beta (E2), progesterone (P4), and luteinizing hormone (LH). The number of preovulatory follicles was 6.8+/-1.4 and 19.6+/-6.6 in M and L gilts, respectively. A lower degree of cumulus expansion and a lower percentage of mature oocytes (TI/M II) was noted in M. Higher LH and E2 peak levels, a longer E2 to LH peak interval, and lower embryo survival was confirmed. Interestingly, despite the lower number of corpora lutea, a higher peripheral blood level of P4 was shown in M than in L gilts. Both diminished follicular development and protracted oocyte maturation may be involved in low fecundity in M, and the present findings may explain these reproductive phenomena.

Improving post-thaw survival of cryopreserved mouse blastocysts by hydrostatic pressure challenge.

The purpose of the experiments was to study the effect of high hydrostatic pressure treatment prior to vitrification to the survival of expanded mouse blastocysts. High hydrostatic pressure has been reported to induce the production of "shock proteins" in bacteria, which can provide a possibility of cross-protection to other environmental stresses. The possible beneficial effects of this alleged principle was examined on embryo vitrification. First, the behaviour of blastocysts was studied at altered pressure conditions. In the second part of the study, pressure treatment was combined with a cryopreservation protocol. Our results indicate that the survival of pressurized mouse embryos depends on the magnitude and the duration of pressure applied. We demonstrated that a preceding pressure treatment strikingly increases the survival of the frozen blastocysts as well as the speed of resumption of the development, and hatching rate.

Evaluation of spermatological parameters in ochratoxin A--challenged boars.

Exposure to certain mycotoxins has been proved to contribute to fertility problems in pigs. Although ochratoxin A (OA) is one of the most common naturally occurring mycotoxins, there is little data concerning the possible effects of this toxin on sperm quality of boars. After a 4-week control period, animals were given 20 microg OA per os daily for 6 weeks, followed by a 9-week withdrawal period. Serum and seminal plasma were monitored for OA with enzyme-linked immunosorbent assay (ELISA). Spermatozoal motility was measured at 0, 24, 48, 72 and 96 h, and ejaculation volume, initial viability and progressive motility were recorded. Samples of testis and epidydimidis were evaluated histologically. Viability, initial forward motility, and motility after 24h storage were significantly reduced in the experimental group in the withdrawal period only. There were no major histological differences in number and morphology of Leydig cells and epydidimal structures between experimental and control boars. Results of the present study demonstrate that OA may affect sperm production and boar semen quality only after a lag period. Further research is required to elucidate the possibility of a direct or indirect interaction between the toxin and germ cells (spermatogonia).

Survival of rapidly frozen hatched mouse blastocysts.

The objective of the present study was to examine the effect of rapid freezing on the in vitro and in vivo survival of zona-pellucida-free hatched mouse blastocysts. Hatched blastocysts were rapidly frozen in a freezing medium containing either ethylene glycol (EG) or glycerol (G) in 1.5 M or 3 M concentration. Prior to freezing, embryos were equilibrated in the freezing medium for 2 min, 10 min, 20 min or 30 min at room temperature. To freeze them, embryos were held in liquid nitrogen vapour [approximately 1 cm above the surface of the liquid nitrogen (LN2)] for 2 minutes and then immersed into LN2. After thawing, embryos were transferred either to rehydration medium (DPBS + 10% foetal calf serum +0.5 M sucrose) for 10 minutes or rehydrated directly in DPBS supplemented with foetal calf serum. In vitro survival of embryos frozen with EG was higher than those frozen with G. The highest survival was obtained with 3 M EG and 2 min or 10 min equilibration prior to freezing, combined with direct rehydration after thawing. Frozen blastocysts developed into normal foetuses as well as unfrozen control ones did, with averages of 30% (control), 26% (EG) and 15% (G). The results show that hatching and hatched mouse blastocysts can be cryopreserved by a simple rapid freezing protocol in EG without significant loss of viability. Our data indicate that the mechanical protection of the zona pellucida is not needed during freezing in these stages.

Comparison of luteinizing hormone and steroid hormone secretion during the peri- and post-ovulatory periods in Mangalica and Landrace gilts.

The objective of this study was to determine plasma concentrations of luteinizing hormone (LH), progesterone (P4) and estradiol-17beta (E2) in Mangalica gilts (M), a Hungarian native breed, and compare them with Landrace gilts (L) during the peri- and post-ovulatory periods. The estrous cycle of gilts was synchronised by Regumate feeding, and ovulation was induced with a gonadotropin-releasing hormone (GnRH) agonist. Blood sampling was carried out via indwelling jugular catheters three times a day and in 2-h intervals during a 16-h period after the GnRH application. The concentrations of LH, E2 and P4 were determined by immunoassays. Gilts of both breeds showed a typical gonadotropin and gonadal hormone secretion pattern. Preovulatory E2 peaks were observed on day 2 (M) and day 4 (L) after the last Regumate feeding. Highest E2 concentration was different between M and L breeds (46.5 +/- 5.7 vs. 26.0 +/- 6.8 pg/ml, P < 0.05). Maximum LH levels measured up to 6 h after GnRH were not different between M and L breeds (11.5 +/- 4.1 vs. 6.6 +/- 2.3 ng/ml). Both LH amounts during surge (41.1 +/- 15.9 vs. 27.5 +/- 6.1 ng/ml) and total over LH release (73.4 +/- 22.2 vs. 50.0 +/- 8.7 ng/ml) did not differ significantly between M and L breeds. P4 concentrations started to rise on day 6 after Regumate feeding and increased significantly from 0.6 +/- 0.3 and 0.7 +/- 0.4 ng/ml to maximal 14.0 +/- 2.4 and 11.3 +/- 2.1 ng/ml in M and L breeds, respectively. Mean P4 secretion was higher in M on days 10-15 (12.9 +/- 2.6 vs. 9.3 +/- 2.2 ng/ml; P<0.05). At the same time the number of corpora lutea was lower in M compared to L (10.3 +/-1.5 vs. 17.8 +/- 5.0, P<0.05). In our experiment, there was no evidence that differences in the secretion of analysed hormones during the peri- and post-ovulatory periods are a possible cause of usually lower fecundity in Mangalica gilts.

Tissue distribution of ochratoxin A as determined by HPLC and ELISA and histopathological effects in chickens.

Ochratoxin A is a common feed contaminant, which may impair animal health and may lead to residues in edible tissues of slaughter animals. To simulate field conditions, broiler chicks were exposed to a total of 0.5 mg ochratoxin A per week for each of 4 weeks. Plasma toxin levels and tissue residues were measured by high-performance liquid chromatography (HPLC) and microplate enzyme-linked immunosorbent assay (ELISA). Results indicate an accumulation in plasma and wide distribution into all organs, with high levels in the liver and the kidney. Microscopical changes that could primarily be associated with toxin exposure were glomerulonephrosis, tubulonephrosis, focal tubular epithelial cell proliferation and multiple, adenoma-like structures in the renal parenchyma. The HPLC and ELISA methods gave similar results for both tissue distribution and depletion. Differences in absolute tissue toxin concentrations obtained by the two methods might be attributed to the different extraction and clean-up procedures, along with antibody specificity. The findings indicate that the dose applied causes subclinical tissue lesions and measurable tissue residues.