Computerized digital image processing on radiographs of canine filariosis.

For objective evaluation in the lung arterial lesions, density histogram revealed by survey thoracic radiographies of fifteen canine filariosis and five normal canine were digitally analyzed, and preparation of pulmonary artery angiogram with inflated-fixed lung, the changes in the histogram and the pulmonary arterial lesion by a soft x-ray examination were compared. In the lung areas affected by filariosis, the density histogram increased the white level and decreased the black level in each part compared to a normal lung. In comparison with the normal parameters, those of the filariosis it were significantly increased in minimum grey level values (Min), maximum grey level values (Max), and the maximum frequency grey level values (Mode) and, it was significantly decreased in maximum frequency values (MaF). The pulmonary arterial lesion of the filariosis showed obvious morphological changes such as in distinction, pruning, angiectasis, and meandering. In the grade of pulmonary arterial lesion, the parameter Min, Max, Mode and MaF were changed significantly. From these results, it was clear that the methods for the lung arterial lesions analysis of x-ray images were confirmed to be highly beneficial in the lung arterial lesions for objective diagnosis.     

Cloning and mapping of three pig acyl-CoA dehydrogenase genes.

To investigate the structure of porcine genes involved in the beta-oxidation of fatty acid, we isolated the short-chain acyl-CoA dehydrogenase (SCAD), medium-chain acyl-CoA dehydrogenase (MCAD), and long-chain acyl-CoA dehydrogenase (LCAD) genes from the pig. The cDNA of SCAD, MCAD and LCAD genes were 1899 bp, 1835 bp 1835 bp and 1704 bp long and coded for 413-aa, 422-aa and 430-aa precursor proteins, respectively. Three genes, SCAD, MCAD and LCAD were mapped to 14p16.2-23.2, 6q32.4-33, and 15q24.2-26.3, respectively.     

Improvement of diagnostic markers for resistance to Globodera pallida and application for selection of resistant germplasms in potato breeding

Occurrence of pale potato cyst nematode, Globodera pallida (Stone) Behrens, was first recorded in Japan in 2015. Among several control measures, cultivation of resistant potato (Solanum tuberosum L.) varieties is the most effective in cost and environmental impact. As no G. pallida-resistant varieties have yet been developed in Japan, great emphasis is being placed on screening of germplasm possessing the resistance and development of the resistant varieties. In this study, we first improved previously reported DNA markers linked to the G. pallida resistance loci (GpaIV^s_adg and Gpa5) and then used these to screen more than 1,000 germplasms to select several candidate germplasms with resistance. We performed inoculation testing on the selected candidates and identified several resistant germplasms to the Japanese G. pallida population. Furthermore, we developed a simultaneous detection method combining three DNA markers linked to G. pallida and Globodera rostochiensis (Wollenweber) Behrens resistance loci. We validated the ability of C237-I marker to select resistant allele of GpaIV^s_adg and predict the presence of resistance in a Japanese breeding population. Resistant germplasms identified in this study could potentially be used to develop G. pallida-resistant varieties. The marker evaluation methods developed in this study will contribute to the efficient development of resistant varieties.     

Habitat use and movement patterns of small (age‐0) juvenile Pacific bluefin tuna (Thunnus orientalis) relative to the Kuroshio

The habitat use of Pacific bluefin tuna (Thunnus orientalis; PBF) in nursery waters off the southern coast of Japan was investigated using archival tags over a 3 year study period (2012–2015), and the data were used to examine the free‐ranging habitat preferences of PBF and the relationship between their horizontal movements and the path of the Kuroshio off the Pacific coast of Japan. The path of the Kuroshio fluctuated seasonally, leading to changes in water temperature that strongly influenced the habitat use of small PBF (2–3 months after hatching). Most PBF were present in coastal waters inshore of the path of the current, and their habitat use changed in response to the distance of the current from the coast. The Kuroshio typically flowed along the coast from summer to autumn, and PBF remained in the coastal waters off Kochi Prefecture during this period. In contrast, PBF quickly moved eastward in winter when the current moved away from the coast. Throughout the winter and spring, the area of habitat use extended widely from the eastern end of the southern coast of Japan (the Boso Peninsula) to the offshore Kuroshio‐Oyashio transition region. These findings suggest that the seasonal habitat use and movement behavior of juvenile PBF are influenced by the distance of the Kuroshio axis from the coast, and the ultimate drivers are likely variations in oceanographic conditions and prey availability along the southern coast of Japan.     

Development of new assessment methods for Acropora coral recruitment using coral settlement devices and holes of marine block

We developed new assessment methods for Acropora coral recruitment using coral settlement devices (CSDs) and holes of a marine block. Both devices were deployed in situ before mass spawning of Acropora corals. The CSDs were sampled after 3–4 months of spawning to measure early recruitment, whereas holes of the marine block were observed underwater or MB was sampled to observe holes after 1 year of spawning to measure the recruitment. By combining the results, we would know the recruitment (visually identified underwater) in reefs, including the amount of early recruitment (identified by stereomicroscopic methods), and the environmental condition of the coral reef. These results would help to predict the future of a reef. Nagura Bay had a high early Acropora recruitment; however, due to competition with algae and sedimentation, the recruitment of juvenile corals was extremely low. The reef would possibly be healthy and sustainable at sites where both the early recruitment and recruitment are high. However, if both are low, the reef will be in a critical condition where urgent restoration will be required.     

Growth performance and physiological condition of F1 amago salmon Oncorhynchus masou ishikawae juveniles obtained from broodstock with selective breeding for growth on a low fish-meal diet

A 10-week feeding trial was conducted to evaluate the growth and physiological condition of F1 amago salmon Oncorhynchus masou ishikawae juveniles obtained from broodstock that had been selected for growth on a low fish-meal (LFM) diet. F1 juveniles from broodstock showing superior or inferior growth on a LFM diet (LFM-S, LFM-I), and those from broodstock showing intermediate growth on a fish meal-based (FM) diet (FM-M), were fed a LFM diet and a FM diet. In fish fed the LFM diet, growth of LFM-S was significantly better than FM-M at 3 weeks; however, this superiority in growth disappeared at the end of the 10-week trial. Growth of LFM-I was significantly lower than LFM-S throughout the feeding trial. Growth of fish fed the FM diet was not significantly different among groups, and higher than fish fed the LFM diet. Improvements in the blood hemoglobin concentration and morphological conditions of the distal intestine and liver were observed in LFM-S fed the LFM diet. Although the effect of selective breeding was limited in F1 amago salmon, the present results suggest that continuous selection for growth with a LFM diet has potential as a method to adapt to the limited supply of fish meal.     

The membrane‐type estrogen receptor G‐protein‐coupled estrogen receptor suppresses lipopolysaccharide‐induced interleukin 6 via inhibition of nuclear factor‐kappa B pathway in murine macrophage cells

The female sex hormone estrogen exerts anti‐inflammatory effects. The G‐protein‐coupled estrogen receptor (GPER) has been recently identified as a novel membrane‐type estrogen receptor that can mediate non‐genomic estrogenic effects on many cell types. We previously demonstrated that GPER inhibits tumor necrosis factor alpha‐induced expression of interleukin 6 (IL‐6) through repression of nuclear factor‐kappa B (NF‐κB) promoter activity using human breast cancer cells. Although several reports have indicated that GPER suppresses Toll‐like receptor‐induced inflammatory cytokine expression in macrophages, the molecular mechanisms of the inhibition of cytokine production via GPER remain poorly understood. In the present study, we examined GPER‐mediated inhibition of IL‐6 expression induced by lipopolysaccharide (LPS) stimulation in a mouse macrophage cell line. We found that the GPER agonist G‐1 inhibited LPS‐induced IL‐6 expression in macrophage cells, and this inhibition was due to the repression of NF‐κB promoter activity by GPER. G‐1 treatment also decreased the phosphorylation of inhibitor of κB kinases. Among the mitogen‐activated protein kinases, the phosphorylation of c‐jun N‐terminal kinase (JNK) was increased by G‐1. These findings delineate the novel mechanism of the inhibition of LPS‐induced IL‐6 through GPER‐activated JNK‐mediated negative regulation of the NF‐κB pathway in murine macrophage cells, which links anti‐inflammatory effects to estrogen.     

Involvement of circadian clock in crowing of red jungle fowls (Gallus gallus)

The rhythmic locomotor behavior of flies and mice provides a phenotype for the identification of clock genes, and the underlying molecular mechanism is well studied. However, interestingly, when examining locomotor rhythm in the wild, several key laboratory‐based assumptions on circadian behavior are not supported in natural conditions. The rooster crowing ‘cock‐a‐doodle‐doo’ is a symbol of the break of dawn in many countries. Previously, we used domestic inbred roosters and showed that the timing of roosters' crowing is regulated by the circadian clock under laboratory conditions. However, it is still unknown whether the regulation of crowing by circadian clock is observed under natural conditions. Therefore, here we used red jungle fowls and first confirmed that similar crowing rhythms with domesticated chickens are observed in red jungle fowls under the laboratory conditions. Red jungle fowls show predawn crowing before light onset under 12:12 light : dim light conditions and the free‐running rhythm of crowing under total dim light conditions. We next examined the crowing rhythms under semi‐wild conditions. Although the crowing of red jungle fowls changed seasonally under semi‐wild conditions, predawn crowing was observed before sunrise in all seasons. This evidence suggests that seasonally changed crowing of red jungle fowls is under the control of a circadian clock.