Germination and appressorium formation of Pyricularia oryzae Cavara can be inhibited by reduced concentration of Blasin®Flowable with carbon dioxide microbubbles

We investigated the possibility to reduce the usage of Blasin^®Flowable (BF), a disinfectant inhibiting the germination and appressorium formation of Pyricularia oryzae Cavara conidia, by using carbon dioxide microbubbles (CO₂MB). Germination was significantly inhibited by 10 000-fold diluted BF solution containing CO₂MB generated by the decompression-type generator compared to CO₂ millibubbles (CO₂MMB) and CO₂MB generated by the gas-water circulating-type generator. Appressorium formation in the 10 000-fold diluted BF solution containing both CO₂MBs was less than that in CO₂MMB. Scanning electron microscopy showed wrinkles and dents on the surface of conidia treated with 5 000-fold diluted BF solution containing both CO₂MBs. Via transmission electron microscopy, we observed the expansion of the vacuole and the intracellular space and bloated or absent lipid granules in the conidia treated with BF solution containing both CO₂MBs. Our results show that inhibition of the conidium germination and appressorium formation of P. oryzae Cavara by 10 000-fold diluted BF solution could be achieved by using the decompression-type CO₂MB.     

Effect of different microbubble generation methods on growth of Japanese mustard spinach

To provide the necessary oxygen for plant growth, the effectiveness of different generating methods of microbubbles (MB), a gas-water circulation type or a pressurization type (pressurization treatment), was investigated by comparing the growth characteristics and morphological observation of Brassica campestris L. var. Komatsuna Matsum. The plant growth in the pressurization treatment were significantly less than those in the circulation treatment at 3 weeks after planting. In plants grown in the pressurization treatment, epidermal cell abrasion and plasmolysis, indicators of cell death, were observed in root tip cells at 3 weeks after planting. The growth inhibition of plant in the pressurization treatment is suggested to be related to the oxidization of Fe in the nutrient solution under super-saturated dissolved oxygen conditions, the generation of hydroxyl radicals from MB, the oxidization of the root tip cell membrane, and osmotic stress to the roots.     

Conventional freezing of in vitro-produced and biopsied bovine blastocysts in the presence of a low concentration of glycerol and sucrose

The purpose of this study was to develop a practical cryopreservation method for in vitro-produced (IVP) and sex-predetermined bovine blastocysts that will be applicable to direct transfer of the post-thaw embryos. Blastocysts were harvested 7 days after IVF and allocated to either an intact or biopsy group. The cryoprotective solution contained 0.7 M glycerol and 0, 0.05 or 0.1 M sucrose. Slow cooling at a rate of -0.5 C/min was terminated at -25, -30, or -35 C, and rapid cooling in liquid nitrogen was followed. After one-step thawing and dilution, the IVP blastocysts were cultured for 3 days to assess their survival. The post-thaw survival rate of intact blastocysts after termination of slow cooling at -30 C in 0.7 M glycerol plus 0.1 M sucrose (96.2%) was significantly higher than that at -25 C in 0.7 M glycerol alone (44.4%). The post-thaw survival rate of biopsied bovine blastocysts after termination of slow cooling at -25 C in 0.7 M glycerol alone (53.8%) tended to be lower than that at -25 C in 0.7 M glycerol plus 0.05 M sucrose (91.3%) or -30 C in 0.7 M glycerol plus 0.1 M sucrose (92.3%). Thus, addition of a small amount of sucrose to 0.7 M glycerol cryoprotective solution shortened the process of slow cooling for both the intact and biopsied bovine embryos. Judged from the survival levels in vitro after thawing and one-step dilution of embryos (>80%), this is an improved method of cryopreservation for subsequent direct transfer of IVP and biopsied bovine blastocysts.     

The novel polymorphism of the beta 3-adrenergic receptor gene and its distribution in domestic pigs and wild boars in Asia

The beta 3‐adrenergic receptor (ADRB3) is a G protein‐coupled receptor that is involved in regulating energy homeostasis. We have studied DNA sequences of porcine ADRB3 to find candidate genetic polymorphisms for economically important growth and performance traits in pigs. Five novel haplotypes derived from the three In/dels and 44 SNPs were identified among domestic pigs and wild boars. Three of them encode non‐synonymous amino acid sequences by five missense polymorphisms and a frameshift by a thymine insertion. The amino acid polymorphic sites were distributed as follows: one substitution was in extracellular loop 1, three substitutions were in intracellular loop3 and one substitution and the deletion of two amino acids were at the carboxyl‐terminal. There was no polymorphism in the transmembrane domains. In addition, we surveyed the allelic frequency of the thymine insertion that cause frameshift in South‐east Asian local pigs, including some commercial breeds and wild boars. This thymine insertion was distributed widely in the domestic pigs and wild boars. The frequencies of this allele were relatively low in Western improved breeds, while they were very common in Asian breeds and wild boars in Asia. This result indicates that this insertion originally occurred in ancient Asian wild boars and then circulated among Asian domestic pigs. This allele also spread over Western breeds, probably through the introgression of Asian pigs into European stocks during the 18th and 19th centuries.     

Effects of the fattening period on the fatty acid composition of fat deposits and free amino acid and inosinic acid contents of the longissimus muscle in carcasses of Japanese Black steers

The effects of the fattening period on carcass characteristics, fatty acid composition of fat deposits, and muscle free amino acid (FAA) and inosinic acid (IMP) contents were evaluated in Japanese Black steers. Ten castrated, 10-month-old calves derived from the same sire were divided into five to be slaughtered at the age of 30 months after a 20-month fattening period (20-month group) and five to be slaughtered at the age of 34 months after a 24-month fattening period (24-month group). Concerning the fatty acid composition of subcutaneous fat, the percentage of palmitoleic acid was higher ( P  < 0.05) in the 24- than in the 20-month group, but no difference was noted in any other fatty acids. For intermuscular fat, no difference was observed in any fatty acids. The percentages of oleic acid and total monounsaturated fatty acid of intramuscular and perinephric fat were higher ( P  < 0.05) in the 24- than in the 20-month group. Of the FAAs in the longissimus thoracis muscle, the threonine and tyrosine contents were lower ( P  < 0.05) in the 24- than in the 20-month group. The IMP content was higher ( P  < 0.05) in the 24- than in the 20-month group, suggesting an effect of prolongation of the fattening period.     

Mitochondrial diversity of native pigs in the mainland South and South-east Asian countries and its relationships between local wild boars

In this study, we analyzed DNA sequence of mitochondrial DNA (mtDNA) control regions on the 130 native domestic pigs and eight wild boars in the mainland South and South-east Asian countries including Bhutan, Cambodia, Laos, Myanmar, and Vietnam. Forty-four haplotypes were found in the 138 individuals, 41 were in the domestic and four were in wild boars. Only one haplotype was shared by domestic and wild population in Bhutan. In other cases, mtDNA of wild boars did not show close affinity to that of the domestic pigs in the same location, indicating that the native domestic pigs in these countries did not originate in the present habitat. Phylogenetic analyzes of mtDNA haplotypes recapitulated several major clusters identified in other studies, but 11 haplotypes were grouped in a new cluster we named MTSEA. In most cases, more than one lineage group were present in a sampling station, indicating that the present indigenous domestic pigs may have multiple origins. The MTSEA haplotypes were present in relatively high frequencies in domestic pigs in the mountainous area of mainland South-east Asia (Cambodia and Laos), with a few found in Myanmar and Bhutan. The distributions of MTSEA haplotypes are in great conformity with the distribution of present-day Mon-Khmer language and indicated the existence of yet another independent domestication. The D2 haplotypes that distribute high frequency (almost 100%) throughout the Chinese breeds were dominant in Bhutan, Myanmar, and Vietnam. These results suggest an existence of human-mediated dispersal of domestic pigs from north to the south during the historical expansion of Sino-Tibetan and Tai peoples. The D3 haplotypes previously reported in north India were found in sympatric domestic and wild pigs in Bhutan. The D3 haplotype is an important proof of independent domestication event and/or great gene flow between wild and domestic pigs in the foot of Himalaya.     

Effects of dietary protein level in the early fattening period on free amino acids and dipeptides in the blood and Longissimus thoracis muscle in Japanese Black steers

The effects of crude protein (CP) concentration in feed using soybean meal as its source in the early fattening period on the levels of free amino acids (FAAs) and dipeptides in the blood, and the levels in the M. longissimus thoracis after slaughter were studied in Japanese Black steers. Sixteen steers were divided into four groups and given feed with a CP content of 12, 14, 16, or 18% of dry matter (DM) from 10 to 20 months old, and they were fed with the same level of CP (13.5–13.9% of DM) until slaughter at 30 months of age. There was no significant difference in the weight gain, carcass weight or marbling score between the groups. Concerning the serum FAA and dipeptide contents at 20 months of age, the alanine, tyrosine and tryptophan levels decreased, while the carnosine (Car) level increased, with increases in the CP level in the feed. Although there were no significant differences in the FAA contents of the Longissimus thoracis muscle between the groups, the Car content decreased with increases in the feed protein level.     

Effects of growth rate during the early fattening period on growth, carcass characteristics and circulating hormones in the different growth hormone genotypes of Japanese black steers

One of the bovine growth hormone (GH) genetic variants is a substitution of leucine (Leu) to valine (Val) at amino acid position 127 of the protein. The GH genotypes of 14 Japanese black steers used in the present study were Leu/Leu (A, n = 7) and Val/Val (B, n = 7). The steers in each genotype group were divided into two groups based on intended growth rate (high, 1.0 kg/day; low, 0.6 kg/day) during 10–17 months of age. The overall mean concentration of plasma GH was higher (P < 0.05) in the A group than in the B group. The serum concentration of insulin‐like growth factor‐I was higher (P < 0.05) in the B group than in the A group. The carcass weight of the A group was greater (P < 0.01) than that of the B group. However, there was no significant difference in carcass weights between the 1.0 kg/day and 0.6 kg/day groups (P > 0.05). The rib thickness of the 1.0 kg/day group was greater (P < 0.05) than that of the 0.6 kg/day group. The crude fat content of longissimus muscle was greater (P < 0.05) for the 0.6 kg/day group compared with the 1.0 kg/day group.