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This paper presents new records and noteworthy data on the following taxa in SE Europe and adjacent regions: brown alga Heribaudiella fluviatilis, red alga Batrachospermum skujae, saprotrophic fungus Gnomonia geranii-macrorrhizi, mycorrhizal fungi Amanita alseides and Russula griseascens, liverwort Ricciocarpos natans, moss Blindia acuta, Leucodon sciuroides var. morensis and Pseudostereodon procerrimus, monocots Allium ampeloprasum, Carex ferruginea and Carex limosa and dicots Convolvulus althaeoides, Fumana aciphylla, Hieracium petrovae, Lamium bifidum subsp. bifidum and Ranunculus fontanus are given within SE Europe and adjacent regions

Summary: A multilocus GWAS was performed to explore the genetic architecture of four growth traits in yak. In total, 354 female yaks for which measurements of body weight (BW), withers height (WH), body length (BL) and chest girth (CG) at weaning were available underwent genotyping with the Illumina BovineHD BeadChip (770K). After quality control, we retained 98 688 SNPs and 354 animals for GWAS analysis. We identified seven, 18, seven and nine SNPs (corresponding to seven, 17, seven and eight candidate genes) associated with BW, WH, BL and CG at weaning respectively. Interestingly, most of these candidate genes were reported to be involved in growth‐related processes such as muscle formation, lipid deposition, feed efficiency, carcass composition and development of the central and peripheral nervous system. Our results offer novel insight into the molecular architecture underpinning yak growth traits. Further functional analyses are thus warranted to explore the molecular mechanisms whereby these genes affect these traits of interest. [ABSTRACT FROM AUTHOR]

The Rosalia longicorn (Rosalia alpina) is a strictly protected saproxylic beetle, widely distributed in Central and Southern Europe and mainly associated with ancient beech forests. To improve knowledge about the conservation status of R. alpina in Italy, available molecular markers (microsatellites and mitochondrial cytochrome c oxidase I(COI)) were tested for the first time on Italian populations. The study was performed in four sampling sites distributed in two areas placed in Northern ("Foreste Casentinesi" National Park) and Central Apennines ("Abruzzo, Lazio and Molise" National Park) where populational data about Rosalia longicorn were collected in the framework of the European LIFE MIPP Project. The genetic relationship among Apennine and Central/South-eastern European populations was explored by a comparison with mitochondrial DNA (mtDNA) data from literature. Microsatellite markers were only partially informative when applied to R. alpina Italian individuals, although providing some preliminary indication on an extensive gene flow among populations from the Apennines and local ongoing processes of genetic erosion. Genetic data are consistent with previous ecological data suggesting that the maintenance of variability in this species could be related to both habitat continuity and preservation of large senescent or standing dead trees in forests. Finally, a peculiar origin of the Apennine populations of R. alpina from a putative "Glacial Refugium" in Italy was inferred through COI data. The high genetic distance scored among the analysed populations and those from Central and South-eastern Europe indicates that the R. alpina deme from Apennine Mountains might represent a relevant conservation unit in Europe. Further genetic analyses will allow assessing other possible conservation units of R. alpina and, thus, defining large-scale conservation strategies to protect this endangered longhorn beetle in Europe. [ABSTRACT FROM AUTHOR]

Milk fat determines the quality of milk and is also a main targeted trait in dairy cow breeding. Recent studies have revealed important regulatory roles of microRNAs (miRNA) in milk fat synthesis in the mammary gland. However, the role of miRNA in bovine mammary epithelial cells (BMEC) remains largely unknown. In this study, we found that the overexpression of miR-130a significantly decreased cellular triacylglycerol (TAG) levels and suppressed lipid droplet formation, whereas the inhibition of miR-130a resulted in greater lipid droplet formation and TAG accumulation in BMEC. MiR-130a also significantly affected mRNA expression related to milk fat metabolism. Specifically, the overexpression of miR-130a reduced the mRNA expression of PPARG, FABP3, PLIN2, and FATP1, whereas the downregulation of miR-130a increased the mRNA expression of PPARG, C/EBPβ, C/EBPα, PLIN2, FABP3, and FATP1. Furthermore, western blot analysis revealed the protein level of PPARG in miR-130a mimic and inhibitor transfection groups to be consistent with the mRNA expression response. Finally, luciferase reporter assays verified that PPARG was the direct target of miR-130a. This study provides the first experimental evidence that miR-130a directly affects TAG synthesis in BMEC by targeting PPARG, suggesting that miR-130a potentially could be used to improve beneficial milk components in dairy cows. [ABSTRACT FROM AUTHOR]

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Fatty acid binding protein 4 (FABP4) is a member of the FABP family of proteins that bind fatty acids and play important roles in fatty acid uptake and intracellular transport. In the present study, we cloned the 5'-regulatory region of bovine FABP4 and identified its transcription initiation sites. Sequence comparative analysis demonstrated amino acids and promoter sequences of FABP4 were highly conservative in mammals. Real-time PCR analysis revealed the products of bovine FABP4 were very highly expressed in subcutaneous adipose tissue. Serial deletion constructs of the bovine FABP4 5'-regulatory region evaluated in a dual-luciferase reporter assay showed that 209 bp upstream from the transcription initiation site was its core promoter. An electrophoretic mobility shift assay combined with a site-directed mutation experiment indicated that peroxisome proliferator activated receptor gamma (PPARγ), CCAAT/enhancer-binding protein β (C/ EBPβ), and sterol regulatory element-binding protein (SREBP) were important transcription factors for bovine FABP4. These results provide an important basis for further understanding the regulation of bovine FABP4. [ABSTRACT FROM AUTHOR]

Japanese Wagyu cattle are well known for their extremely high marbling and lower subcutaneous adipose tissue compared with Angus cattle. However, mechanisms for differences in adipose deposition are unknown. The objective of this paper was to evaluate breed differences in the structure of subcutaneous adipose tissue, adipogenesis, and mitogenesis of stromal vascular (SV) cells between Wagyu and Angus cattle. Subcutaneous biopsy samples were obtained from 5 Wagyu (BW = 302 ± 9 kg) and 5 Angus (BW = 398 ± 12 kg) heifers at 12 mo of age, and samples were divided into 3 pieces for histological examination, biochemical analysis, and harvest of SV cells. Adipogenesis of SV cells was assessed by the expression of adipogenic markers and Oil Red-O staining, while mitogenesis was evaluated by an MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium dromide) test, phosphorylation of extracellular signal-regulated kinase (ERK) and protein kinase B (PKB; AKT). Based on histological analysis, Wagyu had larger adipocytes compared with Angus. At the tissue level, protein expression of peroxisome proliferator- activated receptor γ (PPARG) in Wagyu was much lower compared with that of Angus. Similarly, a lower mRNA expression of PPARG was found in Wagyu SV cells. No significant difference was observed for the zinc finger protein 423 (ZNF423) expression between Wagyu and Angus. As assessed by Oil Red-O staining, Wagyu SV cells possessed a notable trend of lower adipogenic capability. Interestingly, higher mitogenic ability was discovered in Wagyu SV cells, which was associated with an elevated phosphorylation of ERK1/2. There was no difference in AKT phosphorylation of SV cells between Wagyu and Angus. Moreover, exogenous fibroblast growth factor 2 (FGF2) enhanced mitogenesis and ERK1/2 phosphorylation of SV cells to a greater degree in Angus compared with that in Wagyu. Expression of transforming growth factor β 3 (TGFB3) and bone morphogenetic protein 2 (BMP2) in Wagyu SV cells was lower than that of Angus, providing potential clues for breed differences on proliferation of SV cells in these two cattle breeds. The results of this study suggest that subcutaneous adipose-derived SV cells of Wagyu possess a lower trend of adipogenesis but higher mitogenesis compared with those of Angus. [ABSTRACT FROM AUTHOR]

Of all the mammals of the world, the yak lives at the highest altitude area of more than 3000 m. Comparison between yak and cattle of the low-altitude areas will be informative in studying animal adaptation to higher altitudes. To investigate the molecular mechanism involved in meat quality differences between the two Chinese special varieties Qinghai yak and Qinchuan cattle, 12 chemical–physical characteristics of the longissimus dorsi muscle related to meat quality were compared at the age of 36 months, and the gene expression profiles were constructed by utilizing the bovine genome array. Significant analysis of microarrays was used to identify the differentially expressed genes. Gene ontology and pathway analysis were performed by a free Web-based Molecular Annotation System 2.0. The results reveal ~11 000 probes representing about 10 000 genes that were detected in both the Qinghai yak and Qinchuan cattle. A total of 1922 genes were shown to be differentially expressed, 633 probes were upregulated and 1259 probes were downregulated in the muscle tissue of Qinghai yak that were mainly involved in ubiquitin-mediated proteolysis, muscle growth regulation, glucose metabolism, immune response and so on. Quantitative real-time PCR (qRT-PCR) was performed to validate some differentially expressed genes identified by microarray. Further analysis implied that animals living at a high altitude may supply energy by more active protein catabolism and glycolysis compared with those living in the plain areas. Our results establish the groundwork for further studies on yaks’ meat quality and will be beneficial in improving the yaks’ breeding by molecular biotechnology. [ABSTRACT FROM PUBLISHER]

Contents Embryo biotechnology has become one of the prominent high businesses worldwide. This technology has evolved through three major changes, that is, traditional embryo transfer ( in vivo embryo production by donor superovulation), in vitro embryo production by ovum pick up with in vitro fertilization and notably current cloning technique by somatic cell nuclear transfer and transgenic animal production. Embryo biotechnology has widely been used in dairy and beef cattle industry and commercial bovine embryo transfer has become a large international business. Currently, many developed biotechnologies during the period from early oocyte stage to pre-implantation embryos can be used to create new animal breeds and accelerate genetic progression. Based on recent advances in embryo biotechnologies and authors current studies, this review will focus on a description of the application of this technology to beef cattle improvement and discuss how to use this technology to accelerate beef cattle breeding and production. The main topics of this presentation include the following: (i) how to increase calf production numbers from gametes including sperm and oocyte; (ii) multiple ovulation and embryo transfer breeding schemes; (iii) in vitro fertilization and intracytoplasm sperm injection in bovine; (iv) pronuclear development and transgenic animals; (v) sex selection from sperm and embryos; (vi) cloning and androgenesis; (vii) blastocyst development and embryonic stem cells; (viii) preservation of beef cattle genetic resources; and (ix) conclusions. [ABSTRACT FROM AUTHOR]

The present study aimed to investigate the effects of vitamin B supplementation on standard bovine semen quality parameters and anti-oxidative enzyme activities. Vitamin B was supplemented at concentrations of 1.25, 2.5, 3.75 and 5.0 mg/ml to bovine semen cryoprotective medium. The results indicated that the motility and straight line velocity, curvilinear velocity, mean coefficient, velocity of the average path values of sperm supplemented with 2.50 mg/ml vitamin B were significantly higher than that of other groups (p < 0.05). No significant difference was observed for linearity index, lateral head displacement values and the percentage of grade A spermatozoa between the extenders containing 2.50 and 3.75 mg/ml vitamin B (p > 0.05). The percentages of acrosome-intact and plasma membrane-intact spermatozoa were significantly improved (p < 0.05) by supplementing with 2.50 mg/ml vitamin B. The results of biochemical assay revealed that vitamin B supplementation did not cause significant changes in superoxide dismutase levels compared with control (p > 0.05). However, the catalase levels were higher in the treatment supplemented with vitamin B at 2.50 mg/ml, when compared with other groups (p < 0.05). The extender supplemented with vitamin B significantly decreased glutathione peroxidase activity compared with the control (p < 0.05). The supplementation of 3.75 mg/ml vitamin B caused the highest value of glutathione reductase activity, compared with other groups (p < 0.05). In conclusion, the extender supplemented with vitamin B could reduce the oxidative stress provoked by freezing-thawing and improve bovine semen quality. Further studies are required to obtain more concrete results on the determination of lipid peroxidation and antioxidant capacities of vitamin B in cryopreserved bovine semen. [ABSTRACT FROM AUTHOR]

The antioxidant systems of superoxide dismutase (SOD), catalase (CAT), reduced glutathione (GSH), and glutathione peroxidase (GSH–Px) are important in maintaining sperm motility and viability. The purpose of this study was to determine the effects of varyilig doses of trehalose on in vitro semen quality variables and antioxidant activities of frozenthawed bovine semen. The semen samples, diluted with an extender containing trehalose (0, 25, 50, 100, and 200 mM), were evaluated. The extender supplemented with 100 mM trehalose exhibited the greatest percentages of sperm motility, acrosomal membrane integrity, and plasma membrane integrity in comparison with the control group (P < 0.05). No difference was observed for sperm motility between trehalose 50 and 100 mM groups (P > 0.05). Extender supplemented with trehalose did not affect SOD levels. Compared with the other groups, CAT was greater with the supplementation of trehalose at 100 and 200 mM (P < 0.05). The extender supplemented with trehalose had enhanced GSH–Px activity compared with the control group (P < 0.05). However, increasing the doses of trehalose (100, 200 mM) decreased GSH–Px activity, compared with 50 mMtrehalose (P < 0.05). Compared with the other groups, trehalose at the concentration of 25 and 50 mM increased GSH activity (P < 0.05). The application of 200 mM trehalose produced the least amount of GSH activity among all of the groups (P < 0.05). In conclusion, extender supplemented with trehalose reduced the oxidative stress induced by freeze–thaw and improved measures of bovine semen quality. The antioxidant characteristics of trehalose may be related to its effectiveness in membrane cryopreservation. Further studies are required to obtain more concrete results on the determination of lipid peroxidation and antioxidant capacities of trehalose in cryopreserved bovine semen. [ABSTRACT FROM AUTHOR]

Growing evidence has revealed that microRNA are central elements in milk fat synthesis in mammary epithelial cells. A negative regulator of adipocyte fat synthesis, miR-27a has been reported to be involved in the regulation of milk fat synthesis in goat mammary epithelial cells; however, the regulatory role of miR-27a in bovine milk fat synthesis remains unclear. In the present study, primary bovine mammary epithelial cells (BMEC) were harvested from mid-lactation cows and cultured in Dulbecco's modified Eagle's medium/F-12 medium with 10% fetal bovine serum, 5 µg/mL of insulin, 1 µg/mL of hydrocortisone, 2 µg/mL of prolactin, 1 µg/mL of progesterone, 100 U/mL of penicillin, and 100 µg/mL of streptomycin. We found that the overexpression of miR-27a significantly suppressed lipid droplet formation and decreased the cellular triacylglycerol (TAG) levels, whereas inhibition of miR-27a resulted in a greater lipid droplet formation and TAG accumulation in BMEC. Meanwhile, overexpression of miR-27a inhibited mRNA expression of peroxisome proliferatoractivated receptor gamma (PPARG), CCAAT/enhancer- binding protein beta (C/EBPβ), perilipin 2 (PLIN2), and fatty acid binding protein 3 (FABP3), whereas miR-27a downregulation increased PPARG, C/EBPβ, FABP3, and CCAAT enhancer binding protein alpha (C/EBPa) mRNA expression. Furthermore, Western blot analysis revealed the protein level of PPARG in miR-27a mimic and inhibitor transfection groups to be consistent with the mRNA expression response. Moreover, luciferase reporter assays verified that PPARG was the direct target of miR-27a. In summary, these results indicate that miR-27a has the ability to control TAG synthesis in BMEC via targeting PPARG, suggesting that miR-27a could potentially be used to improve beneficial milk components in dairy cows. [ABSTRACT FROM AUTHOR]

Isolation of genomic DNA is a prerequisite for assessment of milk quality. As a source of genomic DNA, milk Somatic cells from milking ruminants are practical, animal friendly, and cost-effective sources. Extracting DNA from milk can avoid the stress response caused by blood and tissue sampling of cows. In this study, we optimized a novel DNA extraction method for amplifying long (>1,000 bp) DNA fragments and used it to evaluate the isolation of DNA from small amounts of milk. The techniques used for the separation of milk omatic cell were explored and combined with a sodium dodecyl sulfate (SDS)-phenol method for optimizing DNA extraction from milk. Spectrophotometry was used to determine the concentration and purity of the extracted DNA. Gel electrophoresis and DNA amplification technologies were used for to determine DNA size and quality. The DNA of 112 cows was obtained from milk (samples of 13 ± 1 mL) and the corresponding optical density ratios at 260:280 nm were between 1.65 and 1.75. Concentrations were between 12 and 45 µg/µL and DNA size and quality were acceptable. The pecific PCR amplification of 1,019- and 729-bp bovine DNA fragments was successfully carried out. This novel method can be used as a practical, fast, and economical mean for long genomic DNA extraction from a small amount of milk. [ABSTRACT FROM AUTHOR]