Phylogenetic analysis of Myanmar indigenous chickens using mitochondrial D-loop sequence reveals their characteristics as a genetic resource

Myanmar indigenous chickens play important roles in food, entertainment, and farm business for the people of Myanmar. In this study, complete mitochondrial D-loop sequences (1232 bp) were analyzed using 176 chickens, including three indigenous breeds, two fighting cock populations, and three indigenous populations to elucidate genetic diversity and accomplish a phylogenetic analysis of Myanmar indigenous chickens. The average haplotype and nucleotide diversities were 0.948 ± 0.009 and 0.00814 ± 0.00024, respectively, exhibiting high genetic diversity of Myanmar indigenous chickens. Sixty-four haplotypes were classified as seven haplogroups, with the majority being haplogroup F. The breeds and populations except Inbinwa had multiple maternal haplogroups, suggesting that they experienced no recent purifying selection and bottleneck events. All breeds and populations examined shared haplogroup F. When 232 sequences belonging to haplogroup F (79 from Myanmar and 153 deposited sequences from other Asian countries/region) were analyzed together, the highest genetic diversity was observed in Myanmar indigenous chickens. Furthermore, Myanmar indigenous chickens and red junglefowls were observed in the center of the star-like median-joining network of 37 F-haplotypes, suggesting that Myanmar is one of the origins of haplogroup F. These findings revealed the unique genetic characteristic of Myanmar indigenous chickens as important genetic resources.     

Improvement of an enzyme-linked immunosorbent assay for equine herpesvirus type 4 by using a synthetic-peptide 24-mer repeat sequence of glycoprotein G as an antigen

To increase the sensitivity of an enzyme-linked immunosorbent assay (ELISA) for equine herpesvirus type 4 (EHV-4) that uses a 12-mer peptide of glycoprotein G (gG4-12-mer: MKNNPIYSEGSL) [4], we used a longer peptide consisting of a 24-mer repeat sequence (gG4-24-mer: MKNNPIYSEGSLMLNVQHDDSIHT) as an antigen. Sera of horses experimentally infected with EHV-4 reacted much more strongly to the gG4-24-mer peptide than to the gG4-12-mer peptide. We used peptide ELISAs to test paired sera from horses naturally infected with EHV-4 (n=40). gG4-24-mer ELISA detected 37 positive samples (92.5%), whereas gG4-12-mer ELISA detected only 28 (70.0%). gG4-24-mer ELISA was much more sensitive than gG4-12-mer ELISA.     

Vulnerability of feline T-lymphocytes to charged particles

An analysis of ionizing radiation-induced damage in peripheral lymphocytes has been employed to predict the prognosis of radiotherapy in terms of toxicity in normal tissues. Therefore, understanding the sensitivity of lymphocytes to high linear energy transfer (LET)-charged particles would be indispensable for utilizing charged particle therapy in veterinary medicine. However, the availability of such information is very limited. This study aimed to compare the radiosensitivity of feline T lymphocytes to gamma-rays (0.2 keV/microm) and 4 different types of charged particles with LET values ranging from 2.8 to 114 keV/microm. It was observed that the relative biological effectiveness, inactivation cross-section, and isodose-induced apoptosis increased in an LET-dependent manner. On the other hand, no difference in apoptosis frequency was observed in the cells exposed to an isosurvival dose of all the radiation types tested. This is the first study that demonstrates the LET dependence of cell killing and apoptosis induction in feline T lymphocytes. Our results suggest that lymphocytes can be effectively used to predict the prognosis of charged-particle therapy in cat patients.     

Structure of the intratesticular duct system for sperm emission in the starspotted smooth-hound Mustelus manazo

We conducted histological observation of male germ cells and reproductive organs of the starspotted smooth-hound Mustelus manazo in Tokyo Bay to reveal any abnormality in male reproductive traits, as part of a study to elucidate the factors causing recent fluctuation in abundance of the population. Spermatogenesis proceeded in spermatocysts from the germinal zone in the ventral part of the testis to the degenerative zone in the dorsal part, where the spermatozoa were conveyed into the ciliated lumina of the attached terminal branches of the intratesticular ducts. The intratesticular ducts were classified from their terminal ends into branch, stem, and collecting tubules. The ducts formed in the germinal zone and grew as the spermatocysts developed. An opening formed through the wall of each of the most mature spermatocysts into a branch tubule; bundles of spermatozoa were evacuated through this opening into the branch and then the stem tubule and subsequently into the collecting tubules in the rete testis and the efferent duct connected to the epididymis. Spermatocysts that were unable to emit sperm because of failure of adhesion to the branch tubules were disorganized in situ, as were their spermatozoa. The collapsed spermatocysts seem to be cleared by hemophagocytosis with lymphocytes and leukocytes, which may have been recruited from the epigonal organ. There were no specific abnormalities in the spermatogenesis or the morphological structure of testes, which suggested that an abnormality of male reproductive traits was not the major cause of the recent fluctuation in the population abundance of this species. Details of the intratesticular duct system for sperm emission to the epididymis are the first findings in elasmobranchs worldwide.     

Metabarcoding of feces and intestinal contents to determine carnivorous diets in red-crowned cranes in eastern Hokkaido,Japan

The red-crowned crane Grus japonensis in Hokkaido, Japan forms a closed population as a residence that is independent of the mainland population. Based on observations of a limited number of individuals as well as cranes in captivity, red-crowned cranes are omnivores and eat fish, worms, insects and plants in their own territories except in winter, when they are fed with dent corn that is supplied in eastern Hokkaido. DNA metabarcoding based on high throughput sequencing was carried out using universal primer sets for cytochrome oxidase subunit I gene. Feces from 27 chicks collected in June and July in the period from 2016 to 2018 and intestinal contents from 33 adult and subadult cranes that were found dead almost throughout year in 2006–2013 in the field in eastern Hokkaido were used. Although compositions varied considerably in the cranes, both insects and fish were found in adults and subadults to the same extents, while insects were predominant in chicks. Both insects and fish were detected in all seasons for adults and subadults. Horse flies, scarab beetles and weevils accounted for the most of the insects regardless of the life stage. Dace, stickleback, flatfish and sculpin were the major fish species in adults, while chicks ate almost only stickleback. The results provide the first comprehensive data on carnivorous diets in wild red-crowned cranes in eastern Hokkaido as basis for conservation of red-crowned cranes, for which the life style and area continue to change.     

Effects of pre-feeding of a corn silage-based supplement on the feed intake, milk production and nitrogen utilization of grazing dairy cows

Four Holstein cows were used to determine the effect of timing of the feeding of a corn silage (CS)‐based supplement on the feed intake, milk production and nitrogen utilization of grazing dairy cows. The cows were fed the supplement 2 h before grazing (pre‐grazing) or immediately after grazing (post‐grazing). Cows were grazed for 5 h per day under a rotational grazing system. There was no difference in the herbage and total feed intake between treatments. The milk protein yield for pre‐grazing tended to be higher than that for post‐grazing, whereas the milk yield did not differ between treatments. The total nitrogen intake for pre‐grazing tended to be higher than that for post‐grazing (P = 0.06). There was no difference in the urinary nitrogen output between treatments, whereas the proportion of urinary nitrogen output : total nitrogen intake for pre‐grazing tended to be lower than that for post‐grazing (P = 0.06). The milk nitrogen output and nitrogen retention for pre‐grazing tended to be higher than that for post‐grazing (milk nitrogen, P = 0.06; nitrogen retention, P = 0.05). Nitrogen utilization of grazing dairy cows was improved by feeding a CS‐based supplement before grazing.     

Soft X-ray observation of water distribution in the stem ofCryptomeria japonica D. Don II: Types found in wet-area distribution patterns in transverse sections of the stem

Using soft X-ray photography, two-dimensional observations were made of the water distribution within the green stem ofCryptomeria japonica D. Don. Variations in the distribution pattern of the wet area in the horizontal plane of the lower stem were described. The distribution patterns showed extreme variation among individual trees, with broad wet-area distribution types appearing, namely, the regularly distributed wet area and the irregularly distributed wet area. We defined five basic types of wet-area distribution patterns on the basis of their regularity or irregularity. It was concluded that the between-tree variation in the wet-area distribution causes the between-tree variation in the mean moisture content of the heartwood. The distribution patterns of the wet area were similar for individual trees within each plus-tree clone.Part of this paper was presented at 46th annual meeting of the Japan Wood Research Society at Kumamoto, April 1996     

Pilot Study to Examine Radial Variation in Annual Ring Width, Density and Shrinkage

The annual ring width, density and shrinkage variation from pith to bark in Chinese fir (Cunninghamia lanceolata) and Boka sugi (Cryptomeria japonicd) were studied and compared. The results show that the ring width decreased sharply from pith to bark for Chinese fir. However, the ring width variation pattern for Boka sugi followed a different way, i.e., the ring width decreased to the fifth ring, increased to the tenth ring, decreased again to the fifteenth, and then increased to the twentieth, where it became constant. The large variations of Boka sugi appeared to show the maintenance of fast growth for many years. The annual ring mean density of Chinese fir increased gradually from pith to bark. However, the density changes for Boka sugi indicated the opposite trend, i.e., the mean density decreased gradually from pith to bark. The former showed a pattern as the same as a pine and a larch, and the latter was often found in a cedar and a cypress. The longitudinal shrinkage in juvenile wood was much hig     

Influence of phosphate-buffered sucrose solution on early graft function in feline renal autotransplantation

Graft perfusion with cold heparinized saline has known to induce ischemia and reperfusion injury in feline kidney transplantation. In this study, the effects of phosphate-buffered sucrose solution and heparinized saline solution on early kidney graft function were compared in feline kidney autotransplantation. Perfusion of grafts with or without hypothermic storage with chilled phosphate-buffered sucrose solution prevented ischemia and reperfusion injury despite a very short ischemic time. The results of our study suggest that phosphate-buffered sucrose perfusion and storage solution should be effective to reduce ischemia and reperfusion injury despite a very short ischemic time in feline kidney transplantation.     

Functions of Chisan dams in large-scale granular mass-flow experiments on Sakurajima volcano pumiceous clasts

We investigated the fundamental behaviour of Chisan (check) dams in response to the presence or absence of impounded sediment with different levels of saturation. Large-scale model flume experiments were conducted by use of a model Chisan dam that had been backfilled with pumiceous clasts taken from the Sakurajima volcano in Kagoshima prefecture, Japan. In these experiments, the miniature dam was situated midway down the flume under different backfill sediment conditions and the basal pore-fluid pressure, normal flow depth, and impact load of the granular mass were carefully monitored. Saturated sediment with a volume of 0.6 m³ was released along the slope of the flume segment at 30° and excess pore-fluid pressure was generated, corresponding to a sediment velocity of approximately 4.8 m s^?1. Direct collision of the sediment with the Chisan dam meant the peak impact loads of granular mass against the back surface of the dam exceeded 157 N in experiments without backfill sediment but were as low as 32.1 N in experiments with sediment. Accordingly, placing the backfill sediment substantially reduced peak impact loads. The sediment captured by the Chisan dams was also calculated. For experiments using saturated backfill sediment, the dam captured only 35 % of the material, indicating that most of the granular mass passed over the dam. However, in experiments without backfill sediment or with unsaturated backfill sediment, over 90 % was captured. Although Chisan dams with unsaturated backfill sediment lacked pocket space, almost equivalent granular masses were trapped in these experiments. On the basis of these results, we suggest that Chisan dams with unsaturated backfill sediment could effectively counter debris flow, reducing sediment loads along the streams draining Sakurajima volcano.