History of Mangalica pig is dated back to the first half of 19th century, however some former information is available of Mangalica ancestors in the 18th century. This fatty pig breed attained its’ typical characteristics till the last quarter of 19th century and these features are still existing. Since Mangalica pigs were nearly extinguished after 1950’s the relevant scientific and practical knowledge also stepwise reduced. In the present overview we compare in detail the ancient and modern information on Mangalica reproduction. First estrous of Mangalica gilts was described at around age of 10 to 11 month. Actually, Mangalica gilts can come in heat as early as 7 to 8 months of age and, therefore can have their first farrowing at 11 to 12 months of age. It was reported that several dam lines had quite high litter-size (8-9 piglets) at the beginning of 20th century, however due to different selection scheme these lines disappeared. Fortunately, nowadays we can identify again such more prolific sows. Throughout comparison of old and modern knowledge of Mangalica reproductive biology we should conclude that respectful incorporation of valuable ancient knowledge has same importance as the newest data collected by the most sophisticated instrumentation and methods. Synthesis of them is necessary for the preservation of original genetic value and the market oriented production.

A história do porco Mangalica remonta à primeira metade do século XIX, no entanto, há informação sobre os seus ascendentes no século XVIII. Este porco tipo “gordura” chegou às suas características típicas no último quarto do século XIX sendo que estas ainda se mantêm até hoje. Dado que os porcos Mangalica estavam praticamente extintos após os anos 50’s do século passado também foi reduzida a informação científica e conhecimento prático sobre a raça. Nesta revisão comparamos em detalhe informação antiga e moderna sobre a reprodução dos suínos Mangalica. Descrições antigas relatavam o primeiro cio das marrãs Mangalica aos 10-11 meses de idade. Efetivamente, as fêmeas Mangalica podem ter cios aos 7-8 meses de idade e, portanto, podem ter o seu primeiro parto com 11-12 meses. Há relatos que várias linhas “mãe” tinham ninhadas grandes (8-9 leitões) no início do século XX, no entanto, por causa de diferentes esquemas de seleção essas linhas desapareceram. Afortunadamente, hoje podemos identificar novamente essas porcas mais prolíficas. Através da comparação do conhecimento antigo e atual da biologia reprodutiva da Mangalica devemos concluir que a incorporação do conhecimento antigo válido tem a mesma importância que os novos dados obtidos com metodologias e instrumentos mais sofisticados. A sua síntese é necessária para a preservação do valor genético original e para a produção orientada para o mercado.

A fast and reliable diagnostic system has been developed for the detection of Mangalica meat in foods. This qualitative test is based on a recombinase polymerase amplification which can be performed on the field, in situ, where it may be necessary to determine Mangalica content in food products at once. The required equipments for the procedure are pipettes, a portable homogenizer and a portable thermostat. DNA amplification is carried out at a constant temperature, and the detection is based on antibody reaction. The detection limit is one copy of the target sequence in 1 μl reaction volume. The test can be used for uncovering falsification of local brands on the spot within a very short (25–45 min) period of time. The present approach can be adopted for the detection of other food ingredients, if the species-specific target DNA sequence is known, e.g. in case of chicken, turkey, horse, and cattle.

in liquid nitrogen on post-warming survival of vitrified porcine oocytes ○ Tamas SOMFAI1,Istvan EGERSZEGI2,Nguyen Thi MEN3,Hiroyuki KANEKO4, Junko NOGUCHI4,Jozsef RATKY2,Naomi KASHIWAZAKI5,Kazuhiro KIKUCHI4 1NARO Institute of Livestock and Grassland Science,2NARIC-Research Institute for Animal Breeding, Nutrition and Meat Science, Hungary,3University of Tsukuba,4National Institute of Agrobiological Sciences,5Azabu University

The influence of melatonin implantation on scrotal circumference, semen quantitative and qualitative parameters, plasma melatonin as well as basal and provoked testosterone concentration was evaluated in Black Racka rams in the non-breeding season. Twelve rams were used in the 60-day-long trial; six of them were implanted (M) with Melovine® (Ceva, Libourne, France) subcutaneously on day 0 (10th May) and 30 days later again, while the other six rams remained untreated (C). Scrotal circumference (SC) was measured and semen was collected weekly and assorted to two 30-day-long periods. Blood samples were collected on days 0, 30 and 60 and basal testosterone level and GnRH-induced testosterone response were evaluated. Ejaculate volume (VOL) increased in both groups from the minimum value at the beginning of the trial (0.52±0.08ml vs. 0.44±0.06ml) to the maximum at 60 days after the first implantation (0.82±0.07ml vs. 0.73±0.05ml, P<0.05). Significant differences were found in SC, VOL and total spermatozoa number from day 30 to day 60 after the first melatonin implantation between the treatment groups (P<0.05). The statistical analyses revealed no difference between treated and control groups regarding total motility % (TM%) and progressive motility % (PM%) at the first implantation and the last sampling. However, at the second implantation melatonin treatment had a positive effect on TM% (P<0.05) and PM% (P<0.001) compared to group C. Basal testosterone (Tb) concentrations were not different between groups at the beginning of the treatment; nevertheless, 60 days later a significantly elevated Tb level was measured in group M compared to C (P<0.05). Testosterone response after the GnRH test was not different between groups at the first and second implantation, while at the third sampling elevated Tb was detected and the provoked testosterone concentrations (Tincr) were also higher in group M at 30 and 60min after GnRH treatment (P<0.05). Plasma melatonin level was increased by implantation to 445.3±91.55pmol/L measured at day 30 compared to 92.4±9.15pmol/L on day 0 (P<0.05), and a further increase of melatonin concentration was recorded after the insertion of the second implants at day 60 (699.45±163.91pmol/L; P<0.05). In conclusion, melatonin implantation in the non-breeding season (at the beginning of May) significantly improved the endocrine and exocrine function of testicles and some quantitative as well as quality parameters of the ejaculate in rams of the Hungarian native breed Black Racka.

In vivo-derived embryos at a defined stage of development are often a necessary requirement for ongoing biotechnological applications. Because double fixed-time insemination after ovulation induction is commonly used in pigs to produce embryos, variations in the time of ovulation and fertilization of the ovulated oocytes by spermatozoa mainly of 1 of the 2 inseminations can cause, however, diversities in embryo development. To moderate embryo diversity and to realize a uniform outcome of porcine embryo stages, single laparoscopic fixed-time insemination can be used to minimize embryo diversity. The potential of laparoscopic intrauterine insemination (LIUI) has been demonstrated in sperm-mediated gene transfer (Fantinati et al. 2005) and evaluation of sperm migration (Brüssow et al. 2006, 2011). The aim of the present study was to analyze the development and possible diversity of embryos after LIUI. Forty-eight puberal German Landrace gilts were included in the study. Estrus of gilts was synchronized by 15-day Regu-Mate® (Intervet, Millsboro, DE, USA) feeding and follicle development was stimulated with 850 IU of eCG 24 h after Regu-Mate® treatment. Ovulation was induced by 500 IU of hCG 80 h after eCG treatment. The LIUI was performed 31 h after hCG treatment. To that, ketamine/azaperone-anaesthetized gilts were fixed in a dorsal position, a pneumoperitoneum was produced and 3 trocar cannulas were inserted into the abdomen for optics and instruments. Laparoscopic handling was observed on a television monitor. Each uterine horn was carefully fixed with an atraumatic forceps 10 to 15 cm caudal from the utero-tubal junction and the uterine wall was punctured with a 2.5-mm diameter trocar. A 2.2-mm catheter connected to a syringe was inserted about 3 cm into the uterine lumen and 20 mL of extended, fresh boar semen (32.2 × 106 sperm cells mL–1; 65% motility) was deposited in the lumen. Embryos were surgically flushed from the genital tract on Day 2 and 3, respectively. Altogether, 778 oocytes were recovered (recovery rate 68 ± 17%); 45 of 48 gilts (93.8%) revealed fertilization and 76.1% of the recovered embryos (n = 592) were at the 2- and 4-cell stage. On Day 2 (n = 22 gilts), a higher percentage of gilts displayed only 2-cell embryos compared with both 2- and 4-cell, and only 4-cell embryos (72.2 v. 22.7 and 4.6%, P < 0.05; chi-square test). On Day 3 (n = 23 gilts), there was a shift regarding the embryo stage. The proportion of gilts with 2-cell, 2- and 4-cell, and only 4-cell embryos was 4.3, 0, and 95.7%, respectively (P < 0.05). Results of the present study demonstrate high rates of fertilization and of non-diverse developed embryos after single fixed-time LIUI in gilts. Additionally, these results were achieved after inseminating a 75% lower number of sperm cells per insemination dose. Laparoscopic intrauterine insemination can be suggested as an alternative for insemination of sex-sorted semen where the number of available sperm cells after the sorting procedure is restricted.

Seasonal changes of scrotal circumference, blood plasma testosterone concentration and semen characteristics were studied over a period of 12months in rams of the Hungarian Black Racka breed. Significant differences between seasonal averages of the parameters measured were established in multiple cases. As regards changes in scrotal circumference, the highest average value obtained in September (31.92cm) exceeded the lowest value measured in January (22.08cm) by 44.7%. It has been found that the season significantly affects the testicular dimensions (r=0.87, P<0.01). Significant correlations were found between scrotal circumference and day length (r=0.43, P<0.05) as well as between scrotal circumference and monthly average temperature (r=0.73, P<0.01). The lowest blood plasma testosterone (T) level (2.31±0.14ng/ml) was measured in winter and the highest in autumn (17.81±0.7ng/ml). Significant differences between the seasonal values were observed on multiple occasions. The GnRH-provoked maximal testosterone level (Tincr) was found in autumn, in the breeding season (54.02±3.78ng/ml). A significant correlation was found between T and Tincr on the one hand and scrotal circumference on the other (r=0.62, P<0.01 and r=0.69, P<0.01, respectively). Ejaculate volume tended to increase from spring (0.52±0.03ml) to autumn (0.88±0.03ml). The lowest sperm concentration (4.92±0.13×109/ml) was measured in winter and the highest in summer (6.56±0.21×109/ml) (P<0.001). Total sperm count per ejaculate was the lowest in spring (3.15±0.23×109/ml) and the highest in autumn (5.22±0.45×109/ml) (P<0.001). Sperm motility was at a low level in February (4.42±0.20) and subsequently showed the highest values in the summer months, with similar monthly averages in June–August (5.00±0.00, P<0.001). The percentage of abnormal spermatozoa was the highest in winter (18.48±3.23%) and the lowest in autumn (7.95±1.58%) (P<0.001), and it was significantly higher in February than in other months of the year (22.17±1.61%, P<0.001–P<0.02). The lowest number of deformed spermatozoa was observed in October (6.15±0.72%).

Double fixed-time insemination after ovulation induction is commonly used in pigs to obtain in vivo produced embryos at defined stages of development for downstream biotechnological applications. However, variations in the time of ovulation and fertilisation of the ovulated oocytes by spermatozoa, mainly in one of the inseminations, can cause diversities in embryo development. The aim of the present study was to reduce embryo diversity and to achieve a 'uniform outcome' of porcine embryo stages using single laparoscopic fixed-time insemination (LIUI). Altogether, 48 puberal German Landrace gilts were included in the study. Estrus of gilts was synchronized by 15-day long altrenogest (Regumate®) feeding and follicle development was stimulated with 850 IU eCG 24 h after the final altrenogest application. Ovulation was induced with 500 IU hCG 80 h after eCG. LIUI was performed 31 h after hCG treatment. Gilts under general anaesthesia were fixed in a dorsal position, a pneumoperitoneum was produced and three trocar cannulas were inserted into the abdomen for optics and instruments. Each uterine horn was carefully punctured 10-15 cm caudal from the utero-tubal junction with a 2.5 mm trocar. A 2.2 mm catheter was inserted about 3 cm into the uterine lumen and 20 ml of extended fresh boar semen (32.2 × 106 sperm cells/ml) were injected. Embryos were surgically flushed from the genital tract two (Day 2) and three (Day 3) days after insemination. Altogether, 778 oocytes/embryos were recovered (recovery rate 68 ± 17%); 45 of 48 gilts (93.8%) revealed fertilisation and 76.1% of the recovered embryos (n = 592) were at the 2- and 4-cell stage. On Day 2 (n = 22 gilts), a higher percentage of gilts (72.7%, P < 0.05) displayed only 2-cell embryos compared with gilts which had 2- and 4-cell (22.7%), or only 4-cell embryos (4.6%). On Day 3 (n = 23 gilts), the proportion of gilts with 2-cell, 2- and 4-cell, and only 4-cell embryos shifted to 4.3%, 0% and 95.7%, respectively (P < 0.05). The results of the present study demonstrate high rates of fertilisation and homogenously developed embryos after single fixed-time laparoscopic intrauterine insemination in gilts. Additionally, these results were achieved by inseminating a 60% lower number of sperm cells per insemination dose compared to usual doses used for transcervical insemination. In conclusion, LIUI can be recommended for the in vivo production of embryos in a homogeneous developmental stage, and also as an alternative method for low-dose insemination.

The genetic relationship between 195 Mangalica and 79 non-Mangalica pigs was studied using mitochondrial D-loop SNP genotyping. Altogether, 35 polymorphic sites and 27 haplotypes were identified. Of the haplotypes, eight and 16 are Mangalica and non-Mangalica specific, respectively, while three contain both Mangalica and non-Mangalica individuals. Genetic distance values and phylogenetic analysis indicate that Mangalica individuals are very closely related, and five haplotypes represent approximately 92% of the Mangalica pigs involved in the study, thus determining the major maternal lineages. In contrast to previous microsatellite studies, individuals of Mangalica could not be distinguished as three separate breeds using mtDNA genotyping. Comparing modern and archaeological mtDNA sequences revealed that present day Mangalica is related to pigs that lived in the Carpathian basin where postulated ancestors of Mangalica also lived. This is the first DNA-based genetic evidence to support the described breeding history of Mangalica.

Nowadays there are growing interest for breeding and maintaining indigenous farm animals. Characterization of the breeds means description of their phenotype, molecular genetic background, productive and reproductive performance. Aim of the study was to collect preliminary data about reproductive performance of Hungarian Black Racka ewes and analyze how year and age of the ewes effect on it. Data were collected from an experimental flock of Black Racka sheep during 2007-2010. Ewes were introduced to harem mating with rams for 6 weeks long in season. The further parameters were recorded; number of ewes and pregnant animals after ultrasound check (US), number of lambing, singleton, twin and triplet lambs, birth weight and age of the ewes. Data were analyzed by SPSS 15 Software for Windows. Altogether 163 ewes were introduced to harem mating, from which 90-93% were pregnant after US. 152 ewes lambed without any problem with an average 1.2 prolificacy. It was born 124 singleton, 26 twin (52) lambs and 2 triplets (6). The male-female lambs’ ratio was 47.3:52.7. The mean birth weight was 3.47±0.62 kg. The birth weight of the lambs did not differ between years and age of the ewes; however, in birth weight there was significant difference with relation of birth type (P<0.05). Significant difference was also observed in birth type related with years. Further data should be collected to give more profound description of the breed and effect of year and age of the ewes on it.

We investigated survival, meiotic competence, cytoplasmic maturation, in vitro fertilization, and development of immature porcine (Sus scrofa) oocytes cryopreserved by a modified solid surface vitrification protocol. Cumulus-oocyte complexes (COCs) collected from follicles 3 to 6mm in diameter in abattoir-derived ovaries of prepubertal gilts were either vitrified (Vitrified group), subjected to cryoprotectant treatment (CPA group), or used without any treatment (Control group). Oocyte viability was assayed by staining with fluorescein diacetate. Live oocytes were matured in vitro and their meiotic progression investigated by nuclear staining. In a series of experiments, the glutathione (GSH) content of in vitro-matured oocytes and viability of cumulus cells were assayed simultaneously. The in vitro-matured oocytes were also fertilized and cultured in vitro to assess their ability to be fertilized and to develop to the blastocyst stage, respectively. The proportion of viable oocytes in the Vitrified group was significantly lower than that in the CPA and Control groups (27.7%, 90.4%, and 100%, respectively). Among the three groups, there were no differences in meiotic competence, cumulus viability, and GSH levels at the end of in vitro maturation. Fertilization parameters (i.e., rates of male pronucleus formation, monospermy, and second polar body extrusion) were also similar among groups. However, comparison of the developmental abilities of oocytes in the Vitrified, CPA, and Control groups revealed that the Vitrified group had a significantly reduced ability to undergo first cleavage (34.4%, 63.3%, and 69.0%) and to develop to the blastocyst stage (5.1%, 25.5%, and 34.6%). The mean total cell numbers in blastocysts after 6 d of culture were not significantly different among the Vitrified, CPA, and Control groups (40.3, 42.8, and 43.4). In conclusion, despite low survival rates and impaired development in the Vitrified group, meiotic competence, cytoplasmic maturation, and subsequent fertilization characteristics of surviving germinal vesicle oocytes were unaffected by vitrification, and high-quality blastocysts were produced from vitrified immature oocytes.