Expression and localization of anti-Müllerian hormone and its receptors in bovine corpus luteum.

Although anti-Müllerian hormone (AMH) is involved in the regulation of granulosa cell function in female animals, its role in tissues other than ovarian follicles remains poorly understood. It has also been suggested that cows with high circulating AMH concentrations have increased fertility; however, the mechanism has not been elucidated. This study was conducted to identify the presence of the AMH-signaling system and its target cells in the bovine corpus luteum formed from an ovulated follicle. Immunoblotting revealed that the proteolytically cleaved C-terminal region in AMH (AMHC), a biologically active peptide, was present in trace amounts in the early corpus luteum and significantly increased during the mid to regressed stages. AMHC and cleaved N-terminal region (AMHN) in AMH generate a noncovalent isoform that improves the activity of AMH signaling. An immunohistochemical analysis revealed that AMHC, AMHN, and type II AMH receptor (AMHR2) were localized to luteal cells during the entire estrous cycle. AMH in the corpus luteum seemed to be newly synthesized since AMH expression was detected. These findings suggest that AMH signaling is involved in the regulation of luteal cell function through an autocrine and post-translational processing mechanism. The level of AMHR2 and mRNA expression of AMHR2 and type I AMH receptors (activin-like kinase 2, 3, and 6) were highest in the mid stage. Thus, AMH signaling in the corpus luteum may also be regulated by changes in the receptor levels. Since the transforming growth factor-beta superfamily, to which AMH belongs, is a multifunctional polypeptide growth factor, further studies are needed to evaluate whether AMH signaling has a role in facilitating or inhibiting luteal cell functions.

Calving prediction with continuous measurement of subcutaneous tissue glucose concentration in pregnant cows.

To reduce losses of dams and calves due to unfortunate events, such as dystocia and freezing to death, identifying the onset of calving and providing necessary assistance are crucial. Prepartum increase in blood glucose concentration is a known indicator to detect labor in pregnant cows. However, some issues, including the need for frequent blood sampling and stress on cows, must be resolved before establishing a method for anticipating calving using changes in blood glucose concentrations. Herein, instead of measuring the blood glucose concentrations, subcutaneous tissue glucose concentration (tGLU) was measured in peripartum primiparous (n = 6) and multiparous (n = 8) cows at 15 min intervals using a wearable sensor. A transient increase in tGLU was observed in the peripartum period, with peak individual concentrations occurring between 2.8 h before and 3.5 h after calving. tGLU in primiparous cows was significantly higher than that in multiparous cows. To account for individual variations in basal tGLU, the maximum relative increase in the 3-h moving average of tGLU (Max MA) was used to predict calving. Cutoff points for Max MA were established by parity, with receiver operating characteristic analysis predicting calving within 24, 18, 12, and 6 h. Except for one multiparous cow that showed an increase in tGLU just before calving, all cows reached at least two cutoff points and calving was predicted successfully. The time interval between reaching the tGLU cutoff points that predicted calving within 12 h and actual calving was 12.3 ± 5.6 h. In conclusion, this study demonstrated the potential role of tGLU as a predictive indicator of calving in cows. Advancements in machine learning-based prediction algorithms and bovine-optimized sensors will help in increasing the accuracy of calving prediction using tGLU.

Genetic relationships among emu populations in Japanese farms based on mitochondrial and microsatellite DNA polymorphisms.

Emus (Dromaius novaehollandiae) are expected to become a novel poultry species for producing eggs, meat, and oil. In our previous studies, Japanese emu populations were predicted to have reduced genetic diversity through inbreeding. For a sustainable emu industry in Japan, it is necessary to understand the current genetic structure and relationships in dispersed farms. In this study, we investigated the genetic structure and relationships of six Japanese emu farms based on mitochondrial DNA and microsatellite polymorphisms. We analyzed the DNA sequences of the mitochondrial D-loop region in 157 individuals and detected four haplotypes with four nucleotide substitution sites (Hap-a, Hap-b, Hap-c, and Hap-d). Analysis of molecular variance revealed that 43.6% of total variance was "among population," and the FST value was 0.436 with significant genetic differentiation (P < 0.001). In microsatellite analysis, the expected (HE ) and observed (HO ) heterozygosities were 0.53-0.64 and 0.44-0.59, respectively. Phylogenetic trees and STRUCTURE analysis revealed that the six Japanese farmed emu populations could be divided into four genetically differentiated groups. Therefore, we identified genetic resources that may be useful in extending the genetic diversity of Japanese farms and are predicted to contribute to the conservation and reconstruction of populations.

Carcass traits and fat quality of breeding emu (Dromaius novaehollandiae) in Northern Japan.

Characterization of carcass traits and fat quality is important to effectively produce and genetically improve emus. We investigated carcass traits in 309 emus. The meat production of female emus showed a significantly higher value than that of males (P < 0.01). The fat weight of male (9.232 ± 3.156 kg) was larger than that of the female (7.772 ± 2.697 kg). The fat yield (fat weight per kg of body weight) was strongly correlated to body weight (r = 0.79 and r = 0.75 in male and female, respectively). The fat melting points of females and males were 19.19 ± 3.39°C and 19.39 ± 3.39°C, respectively, without significant difference. Since the fat melting point did not correlate to body and fat weights, we predicted that it was an independent trait from body growth and was highly influenced by genetic elements. Percentages of palmitic, stearic, oleic, linoleic, and α-linolenic acids were 22.27 ± 3.50%, 9.37 ± 1.90%, 54.11 ± 5.17%, 13.54 ± 7.80% and 0.71 ± 0.59%, respectively. Among them, linoleic acid contents showed a wide individual difference (range 0.3-19.9%). The oleic/stearic acid ratio showed a negative correlation to the fat melting point. These results suggest that the fat melting point is a good indicator of C18:1/C18:0 ratio in emu fat.

Impaired placentomal interferon signaling as the possible cause of retained fetal membrane in parturition-induced cows.

Although hormonal induction of parturition in cattle results in the successful delivery of healthy calves, the risk of retained fetal membrane is significantly increased. In a previous study, a combination of the long-acting glucocorticoid, triamcinolone acetonide, with a high dose of betamethasone partially normalized the placentomal gene expression during parturition; however, the incidence of retained fetal membrane remained high. This study further explored placentomal dysfunction and aimed to elucidate the mechanism of retained fetal membrane in parturition-induced cows. In this study, transcriptome analysis revealed that enhanced glucocorticoid exposure normalized the expression of a substantial fraction of genes in the cotyledons. In contrast, a significant reduction in the multiple signaling pathway activities, including interferon signaling, was found in the caruncles during induced parturition. Real-time PCR showed that the expression of interferon-tau in the caruncles, but not interferon-alpha or interferon-gamma, was significantly lower in induced parturition than spontaneous parturition. Interferon-stimulated gene expression was also significantly decreased in the caruncles during induced parturition. These results indicate that interferon signaling could be important for immunological control in placentomes during parturition. Additionally, this suggests that interferon-tau might be a pivotal ligand for interferon receptors in the caruncles. This study revealed that peripheral blood leukocytes in prepartum cows transcribed interferon-tau. Macrophage infiltration in the placentome is known to participate in the detachment of the fetal membrane from the caruncle. Thus, this study raised the possibility that immune cells migrating into the caruncles at parturition may act as a source of ligands that activate interferon signaling.

Development of 49 novel microsatellite markers from Next-generation sequencing data and a robust method for parentage tests in the emu (Dromaius novaehollandiae).

The emu is a useful and new breed of poultry, but their genetic improvement has not advanced yet due to their very recent domestication. Pedigree information is difficult to record because of their complex reproduction system (polyandry). To identify parent-offspring relationships in the emu, parentage test based on polymorphic DNA markers have to be developed. In this study, we isolated more than 25,000 microsatellite (simple sequence repeat, SSR) regions from Next-generation sequencing data via the QDD pipeline and developed 49 SSR markers with polymorphism in the Japanese farmed emu. The dinucleotide motifs, (AC)n, (AT)n and (AG)n, were the most frequently detected and were found on 10,167 (38.55%), 8,114 (30.76%) and 4,796 (18.18%) contigs, respectively. Forty-nine novel SSR markers were characterized in 20 individuals and showed NA ranged from 2 to 12, with an average of 4.2. HE/HO ranged from 0.389/0.071 to 0.702/1.000 with an average of 0.601/0.515. PIC value ranged from 0.059 to 0.886 with an average of 0.528, and 17 of 49 markers showed a higher polymorphism than 0.500. Thirty-four individuals were genotyped using 12 markers, and CERVUS simulations based on genotype showed that parents of all offspring were identified with 0.9995-1.0 probability. Thus, 49 novel SSR markers and a robust method for parentage test for the Japanese emu were developed.

Mitochondrial DNA variations in Japanese farmed emu populations.

The emu (Dromaius novaehollandiae) is a new poultry. In this study, we investigated the haplotype composition of mitochondrial DNA among emu populations farmed in Japan. We sequenced the D-loop region in 109 individuals, and detected four substitution sites and three haplotypes (Hap-a, -b, and -c). Hap-a was the most frequently observed haplotype in the Japanese populations. Although Hap-c was a rare haplotype in not only Japanese but also Australian populations, it was detected with high frequency in the Japanese farmed population. The AMOVA indicated that 9% of total variance was "among population". The FST value was 0.087 and genetic differentiation was significant (P<0.01). These results may contribute to conserving the genetic resources available for the Japanese emu industry.

Development and characterization of ten novel microsatellite loci for the emu (Dromaius novaehollandiae) and genetic diversity of Japanese farm populations.

The emu (Dromaius novaehollandiae) is a useful poultry animal farmed for fat, meat, and eggs. Genetic structure and relationships among farmed emu populations in Japan are unknown and the number of microsatellite markers for genetic analysis of the emu is insufficient. In this study, we isolated 16 microsatellites from the emu genome and developed ten new microsatellite markers. These microsatellite markers were used to characterize three farm emu populations in Japan. The number of alleles ranged from 3 to 13 and the expected (HE) and observed heterozygosity (HO) of these microsatellite loci was 0.187-0.802 and 0.179-0.647, respectively. The polymorphic information content ranged from 0.176 to 0.786. Positive inbreeding coefficient (FIS) values were detected in all tested populations, and they ranged from 0.027 to 0.540. These results suggest that farm populations of the emu in Japan resulted from inbreeding. The fixation index (FST) values ranged from 0.026 to 0.061, and phylogenetic trees and population structure analysis confirmed no definitive genetic differentiation among the three populations. Therefore, these populations are at a relatively low level of genetic differentiation at present. The microsatellite markers developed in our study can be utilized for genetic analysis and preservation of genetic resources in the emu.

Data on chemical compositions and fermentation quality of silages made from low-market-value vegetables supplemented with potato protein concentrate, a byproduct of starch production.

This data article reports the chemical compositions (protein, fat, fiber, ash, lactic acid, acetic acid, propionic acid, butyric acid and valeric acid) and fermentation quality, represented by V-value determined from the proportion of ammonia nitrogen in total nitrogen and volatile fatty acid contents, in silages prepared from low-market-value vegetables (carrot roots, cabbage leaves, and radish leaves). Potato protein concentrate, a byproduct of starch production from potato tuber, was used to supplement the protein contents in the silages. The first type of silage was produced by fermentation of a mixture of wheat bran and either carrot, cabbage, or radish without supplemental potato protein. The second type of silage was produced by fermentation of a mixture of wheat bran and either carrot, cabbage, or radish with supplemental potato protein. The third type of silage was produced by mixing the first silage type with unfermented potato protein. Chemical compositions and fermentation quality of the three silage types are provided in table formats.

Chemical characterization of the milk oligosaccharides of some Artiodactyla species including giraffe (Giraffa camelopardalis), sitatunga (Tragelaphus spekii), deer (Cervus nippon yesoensis) and water buffalo (Bubalus bubalis).

Mammalian milk/colostrum usually contains oligosaccharides along with the predominant disaccharide lactose. It has been found that the number and identity of these milk oligosaccharides varies among mammalian species. Oligosaccharides predominate over lactose in the milk/colostrum of Arctoidea species (Carnivora), whereas lactose predominates over milk oligosaccharides in Artiodactyla including cow, sheep, goat, camel, reindeer and pig. To clarify whether heterogeneity of a variety of milk oligosaccharides is found within other species of Artiodactyla, they were studied in the milk of giraffe, sitatunga, deer and water buffalo. The following oligosaccharides were found: Neu5Ac(α2-3)[GalNAc(ß1-4)]Gal(ß1-4)Glc (GM2 tetrasaccharide), and Gal(α1-3)Gal(ß1-4)Glc (isoglobotriose) in giraffe milk; Neu5Ac(α2-3)Gal(ß1-4)Glc (3'-SL), Neu5Ac(α2-6)Gal(ß1-4)Glc (6'-SL), Gal(α1-4)Gal(ß1-4)Glc (globotriose) and isoglobotriose in sitatunga colostrum; Gal(ß1-3)Gal(ß1-4)Glc (3'-GL), Gal(ß1-6)Gal(ß1-4)Glc (6'-GL), isoglobotriose, Gal(ß1-4)GlcNAc(ß1-3)Gal(ß1-4)Glc (lacto-N-neotetraose, LNnT), Gal(ß1-4)Glc-3'-O-SO3 (3'-O-lactose sulphate) in deer milk; 3'-GL, isoglobotriose and Gal(ß1-3)Gal(ß1-3)Gal(ß1-4)Glc (3',3″-digalactosyllactose, DGL) in water buffalo colostrum. Thus it was shown that the milk oligosaccharides are heterogeneous among these Artiodactyla species.

Detection of hydrolytic activity of trypsin with a fluorescence-chymotryptic peptide on a TLC plate.

To find a new trypsin-like enzyme, a simple assay method of the hydrolysis activity for trypsin has been found. We used 6-aminoquinolyl-N-hydroxysuccinimidyl carbamate (AQC) in the peptide labeling as a substrate for the trypsin-like peptidase in this study. The peptidase activity of trypsin was detected by using an AQC-chymotryptic peptide (AHP1) obtained from bovine hemoglobin. This showed that the substrate specificity of trypsin-like peptidase was distinguishable from that of the others by this procedure, and the method was used extensively in cases of various trypsin inhibitors with no significant interference from the concomitant.

Isolation and characterization of a cysteine protease of freesia corms.

A protease, freesia protease (FP)-A, was purified to electrophoretic homogeneity from regular freesia (Freesia reflacta) corms in harvest time. The Mr of FP-A was estimated to be 24 k by SDS-PAGE. The optimum pH of the enzyme was 8.0 using a casein substrate. These enzymes were strongly inhibited by p-chloromercuribenzoic acid but not by phenylmethane-sulfonylfluoride and EDTA. These results indicate that FP-A belongs to the cysteine proteases. The amino terminal sequence of FP-A was similar to that of papain, and the sequences was regarded to the conservative residues of cysteine protease. From the hydrolysis of peptidyl-p-NAs, the specificity of FP-A was found to be broad. It was thought that FP-A was a new protease from freesia corms.