Template-Type: ReDIF-Article 1.0 Author-Name: Rosario Rullo Author-Workplace-Name: Institute for the Animal Production Systems in the Mediterranean Environment, Napoli, Italy Author-Name: Aldo Di Luccia Author-Workplace-Name: Department of Agricultural, Food and Environmental Sciences, University of Foggia, Foggia, Italy Author-Name: Elena Ciani Author-Workplace-Name: Department of Biosciences, Biotechnologies and Biopharmaceutics, University of Bari "Aldo Moro", Bari, Italy Author-Name: Elisa Pieragostini Author-Workplace-Name: Department of Emergency and Organ Transplantation, University of Bari "Aldo Moro", Bari, Italy *Corresponding autor: elena.ciani@uniba.it Title: The haemoglobin subunits alpha and beta: Old and new genetic variants in the Italian Mediterranean buffalo Abstract: Haemoglobin (HB), the most widely distributed respiratory pigment in the animal kingdom, is among the best characterized oxygen-binding proteins, both at functional and molecular level. However, very little information is available about the genomic features of HB in river buffalo (Bubalus bubalis), even though there are reports in literature confirming the presence of interesting polymorphisms at the protein level in Mediterranean buffalo. We hence address the characterization of exonic as well as intronic nucleotide polymorphism in the haemoglobin subunit alpha and beta in a set of nine Italian Mediterranean buffaloes exhibiting different HB phenotypes. The nine buffaloes were selected from a random set of 398 samples, previously analysed for their HB protein polymorphism, in order to account for both globin variants and the evolution of intron variability within the most common domesticated species of the family Bovidae. All four sequenced clones of the subunit alpha were 1311 bp, whereas the length of the five different sequenced clones of the subunit beta ranged from 1841 to 1960 bp, due to an insertion of 119 nucleotides. Six polymorphic sites were detected in the four amplicons of alpha subunit. Among them, two variations concern exclusively haplotype A, while four sequence variations were found to be specific to haplotype B. Several variations, both in exonic and intronic regions, were detected in the B. bubalis subunit beta. In conclusion, the nucleotide sequence variants observed in this work substantiate the known haemoglobin protein polymorphisms, and an updated protein nomenclature is provided here. In addition, we observed a high sequence similarity in the overall pattern of variation in the haemoglobin subunits, possibly the results of a concerted evolution, with relatively more extensive gene homogenization in river buffalo than in other ruminant species. Keywords: Bubalus bubalis, haemoglobin subunit variation, biodiversity, transposable elements, gene duplication Journal: Czech Journal of Animal Science Pages: 279-290 Volume: 64 Issue: 7 Year: 2019 DOI: 10.17221/14/2018-CJAS File-URL: http://cjas.agriculturejournals.cz/doi/10.17221/14/2018-CJAS.html File-Format: text/html X-File-Ref: http://agriculturejournals.cz/RePEc/caa/references/cjs-201907-0001.txt Handle: RePEc:caa:jnlcjs:v:64:y:2019:i:7:id:14-2018-CJAS Template-Type: ReDIF-Article 1.0 Author-Name: Shanshan Xie Author-Workplace-Name: Institute of Animal Sciences, Chinese Academy of Agricultural Sciences, Beijing, P.R. China Author-Name: Lili Qian Author-Workplace-Name: Institute of Animal Sciences, Chinese Academy of Agricultural Sciences, Beijing, P.R. China Author-Name: Chunbo Cai Author-Workplace-Name: Institute of Animal Sciences, Chinese Academy of Agricultural Sciences, Beijing, P.R. China Author-Name: Shengwang Jiang Author-Workplace-Name: Institute of Animal Sciences, Chinese Academy of Agricultural Sciences, Beijing, P.R. China Author-Name: Gaojun Xiao Author-Workplace-Name: Institute of Animal Sciences, Chinese Academy of Agricultural Sciences, Beijing, P.R. China Author-Name: Ting Gao Author-Workplace-Name: Institute of Animal Sciences, Chinese Academy of Agricultural Sciences, Beijing, P.R. China Author-Name: Xiang Li Author-Workplace-Name: Institute of Animal Sciences, Chinese Academy of Agricultural Sciences, Beijing, P.R. China Author-Name: Wentao Cui Title: Safety evaluation of myostatin-edited Meishan pigs by whole genome resequencing analyses Abstract: Genome editing technology can make specifically target genomic modifications, resulting in site specific DNA insertion, deletion or replacement in the genome of an organism. We have recently produced genetically engineered (GE) Meishan pigs containing a ZFN-edited myostatin (MSTN) loss-of-function mutation that leads to a clear "double muscle" phenotype as observed for Belgian cattle. In this study, whole genome resequencing was used as an approach to evaluate the safety risk, if any, associated with the introduction of a ZFN-edited myostatin (MSTN) loss-of-function mutation in a local pig breed, the Meishan pigs. The results of resequencing analyses show that the effective data from pigs of wild-type group and MSTN-edited GE group is greater than 99%. The 1× coverage rate is > 98%, and the 4× coverage rate is > 96%. The genetic variation on each chromosome is close to 1. From this whole genome resequencing study, our results demonstrated that 99.7% of single nucleotide polymorphisms (SNPs) are the same in the same genetic variation from both wild-type group and MSTN-edited GE group, implying genomic sequence variations are highly similar between the two groups of pigs. Keywords: MSTN, gene editing, genome resequencing, genetic variation, comparative analysis Journal: Czech Journal of Animal Science Pages: 291-299 Volume: 64 Issue: 7 Year: 2019 DOI: 10.17221/7/2018-CJAS File-URL: http://cjas.agriculturejournals.cz/doi/10.17221/7/2018-CJAS.html File-Format: text/html X-File-Ref: http://agriculturejournals.cz/RePEc/caa/references/cjs-201907-0002.txt Handle: RePEc:caa:jnlcjs:v:64:y:2019:i:7:id:7-2018-CJAS Template-Type: ReDIF-Article 1.0 Author-Name: Amparo Roca Author-Workplace-Name: Department of Agro-Food Technology, University of Miguel Hernández, Orihuela, Spain Author-Name: Gema Romero Author-Workplace-Name: Department of Agro-Food Technology, University of Miguel Hernández, Orihuela, Spain Author-Name: Manuel Alejandro Author-Workplace-Name: Anabel Segura 7, 28108 Alcobendas, Madrid, Spain Author-Name: Raquel Muelas Author-Workplace-Name: Department of Agro-Food Technology, University of Miguel Hernández, Orihuela, Spain Author-Name: José Ramón Díaz Author-Workplace-Name: Department of Agro-Food Technology, University of Miguel Hernández, Orihuela, Spain Title: Milk electrical conductivity in Manchega ewes: Variation throughout milking and relation with mammary gland health status Abstract: The aim of this work was to study the effect of milking fraction and mammary gland health status on the electrical conductivity (EC) of milk from Manchega ewes, considering also the lactation number. To this end, we also studied the relationship of EC with milk macrocomposition, and the relation existing between EC and somatic cell count (SCC). Finally, the use of EC thresholds as a mastitis detection method (sensitivity, specificity, positive (PPV) and negative predictive value (NPV)) was assessed in each of the three fractions: first streams (F1), machine milk (F2) and stripping milk (F3). Milking fraction, mammary gland health status and lactation number had a significant effect on EC and SCC. In the case of EC, the milking fraction caused a more pronounced effect than health status of the glands or number of lactation (F = 19.95, 15.88 and 6.55, respectively; P < 0.5). In SCC, the gland health status caused the most pronounced effect followed by milking fraction and lactation number (F = 112.02, 6.89, and 5.28, respectively; P < 0.05). Changes in the milk composition, especially fat and lactose contents, explained the EC variation to a great extent. For the same EC threshold, specificity and sensitivity varied slightly depending on the milking fraction. NPV above 80% was obtained in the three milking fractions and at all EC thresholds tested, but PPV was only higher than 20% as of the threshold of 4.5, 4.4, and 4.2 mS/cm in F1, F2 and F3, respectively. From the results obtained, we concluded that the algorithm design for mastitis detection in sheep should include those factors affecting the composition and which therefore cause variations in EC, such as milking fraction, individual differences, lactation stage or lactation number. Keywords: mastitis detection, milking fraction, sheep, sensitivity, specificity Journal: Czech Journal of Animal Science Pages: 300-308 Volume: 64 Issue: 7 Year: 2019 DOI: 10.17221/48/2019-CJAS File-URL: http://cjas.agriculturejournals.cz/doi/10.17221/48/2019-CJAS.html File-Format: text/html X-File-Ref: http://agriculturejournals.cz/RePEc/caa/references/cjs-201907-0003.txt Handle: RePEc:caa:jnlcjs:v:64:y:2019:i:7:id:48-2019-CJAS Template-Type: ReDIF-Article 1.0 Author-Name: Michala Hofmannová Author-Name: Josef Přibyl Author-Workplace-Name: Institute of Animal Science, Prague-Uhříněves, Czech Republic Author-Name: Emil Krupa Author-Workplace-Name: Institute of Animal Science, Prague-Uhříněves, Czech Republic Author-Name: Petr Pešek Author-Workplace-Name: Institute of Animal Science, Prague-Uhříněves, Czech Republic Title: Estimation of inbreeding effect on conception in Czech Holstein Abstract: The influence of calculated inbreeding coefficients on the conception of heifers and cows was analysed by a two-trait binary model for conceived/not conceived after each insemination on 677 234 Czech Holstein cows and heifers with 3 248 299 insemination records as the covariable in a statistical model. The data between 1996 and 2014 were analysed. Various effects and their statistical influence on traits were tested by a generalized linear model. Consequently, genetic parameters were estimated by the Gibbs sampling method and used in predicting breeding values using the best linear unbiased prediction by animal model (BLUP-AM). The mean for the conception rate of cows averaged over lactations was 33.7% and for heifers it reached 53.8%. Average inbreeding coefficient increased from 1% in 1996 to almost 5% by 2013 and was in the range of 0-45%. The rate of inbreeding per generation was 0.20%. Although the effect of inbreeding was statistically significant (P = 0.05) for both traits, the proportion of variability explained by the models was relatively low. Estimated genetic parameters were low for both traits. Coefficient of heritability was 2.00% and 1.30% for cows and heifers, respectively, whereas coefficients of repeatability reached 6.09% and 7.08% for cows and heifers, respectively. The random effect of the permanent environment (PE) reached higher values than the additive genetic variance (G) and explained 5.67% and 4.09% of variability for cows and heifers, respectively. A negative impact of inbreeding on heifer and cow conception was observed, whereby every 10% increase in inbreeding coefficient resulted in a conception decline by 2.23%. Calculated Spearman's rank correlation coefficient between estimated breeding values considering and not considering the effect of inbreeding was close to one. Presented results indicate that inbreeding has a negligible influence on the breeding values of conception. The results also indicate that it is not necessary to include inbreeding coefficient in the routine breeding value evaluation of conception rate of heifers and cows. On the other hand, monitoring of inbreeding is necessary to avoid an increase of its rate. Keywords: BLUP, breeding value, dairy cattle, Gibbs sampling, inbreeding depression Journal: Czech Journal of Animal Science Pages: 309-316 Volume: 64 Issue: 7 Year: 2019 DOI: 10.17221/154/2018-CJAS File-URL: http://cjas.agriculturejournals.cz/doi/10.17221/154/2018-CJAS.html File-Format: text/html X-File-Ref: http://agriculturejournals.cz/RePEc/caa/references/cjs-201907-0004.txt Handle: RePEc:caa:jnlcjs:v:64:y:2019:i:7:id:154-2018-CJAS Template-Type: ReDIF-Article 1.0 Author-Name: Leying Zhang Author-Workplace-Name: Department of Animal Science, College of Life Sciences and Food Engineering, Hebei University of Engineering, Handan, P.R. China Author-Name: Chen Zhuang Author-Workplace-Name: Department of Animal Science, College of Life Sciences and Food Engineering, Hebei University of Engineering, Handan, P.R. China Author-Name: Zimo Zhao Author-Workplace-Name: Department of Animal Science, College of Life Sciences and Food Engineering, Hebei University of Engineering, Handan, P.R. China Author-Name: Ning Li Author-Workplace-Name: Department of Animal Science, College of Life Sciences and Food Engineering, Hebei University of Engineering, Handan, P.R. China Author-Name: Jiachen Bai Author-Workplace-Name: Department of Animal Science, College of Life Sciences and Food Engineering, Hebei University of Engineering, Handan, P.R. China Author-Name: Ling Yang Title: Effect of early pregnancy on the expression of progesterone receptor and progesterone-induced blocking factor 1 in ovine liver Abstract: Liver plays important roles in the innate and adaptive immunity, and contributes to the maternal immune adjustments during pregnancy. Progesterone (P4) has key effects on immunomodulation of the maternal uterus during pregnancy. In this study, livers were obtained at day 16 of the estrous cycle and at days 13, 16 and 25 of pregnancy (n = 6 for each group) in ewes. The effects of early pregnancy on the expression of P4 receptor (PGR) and progesterone-induced blocking factor 1 (PIBF1) were analysed through RT-qPCR assay, Western blot and immunohistochemistry analysis. Our results showed that the isoforms of PGR with molecular weights of approximately 60 kDa (PGR60) and 89 kDa (PGR89) were strongly expressed in the livers from pregnant ewes (P < 0.05), but there was no expression of the isoform of PIBF1 with a molecular weight of approximately 55 kDa (PIBF55) on day 13 of pregnancy. The PGR protein was mainly limited to the endothelial cells of the proper hepatic arteries and portal veins, and hepatic cells. In conclusion, the PGR89 and PGR60 were up-regulated, and PIBF55 was down-regulated on day 13 of pregnancy, which may be involved in maternal hepatic immunoregulation during early pregnancy in sheep. Keywords: ewe, immunoregulation, isoform, progesterone Journal: Czech Journal of Animal Science Pages: 317-323 Volume: 64 Issue: 7 Year: 2019 DOI: 10.17221/21/2019-CJAS File-URL: http://cjas.agriculturejournals.cz/doi/10.17221/21/2019-CJAS.html File-Format: text/html X-File-Ref: http://agriculturejournals.cz/RePEc/caa/references/cjs-201907-0005.txt Handle: RePEc:caa:jnlcjs:v:64:y:2019:i:7:id:21-2019-CJAS