In vitro induction of oocyte maturation in the Japanese sea cucumber Apostichopus japonicus by cubifrin and the developmental ability of the eggs

We describe the process of cubifrin-induced in vitro oocyte maturation in the Japanese sea cucumber Apostichopus japonicus, and we propose a procedure for practical hatchery application. Oocyte maturation occurred 40–50 min after stimulation of the ovaries by cubifrin. Concurrently, eggs were released from the ovarian tubules, initially by slow peristalsis, followed by active release after about 60 min. The first and second polar bodies were released from the eggs at 90–100 min and about 140 min, respectively. Cubifrin was more effective in inducing oocyte maturation at 1–100 nM than at 100 pM. Cubifrin at 1 pM–10 nM in rearing seawater had no effect on larval development. Delayed fertilization reduced the developmental ability of eggs. Broodstock could be maintained at 15–16 °C for up to about 50 days without affecting the developmental ability of eggs. Eggs matured in vitro and in vivo were similar in their developmental ability. In conclusion, the in vitro induction of oocyte maturation is effective in Japanese sea cucumber and represents a viable alternative to the induction of spawning by cubifrin injection.     

Novel reovirus isolation from an Ostrich (Struthio camelus) in Japan

An orthoreovirus was isolated from an Ostrich (Struthio camelus) and rapidly identified as orthoreovirus by the rapid determination of viral RNA sequences (RDV) system and electron microscopy. Phylogenetic analysis of the sigma A protein indicated that the isolate belonged to avian species and was closely related to chicken orthoreovirus strain 138. The results of the present study indicated that an ostrich orthoreovirus is slight different from other chicken orthoreoviruses and provided evidence of diversity among avian orthoreoviruses. To our knowledge, this is the first genetic report of an orthoreovirus isolated from an ostrich.     

Rapid visualization of mammary gland tumor lesions of dogs using the enzyme-activated fluorogenic probe; γ-glutamyl hydroxymethyl rhodamine green

Since gamma-glutamyl transpeptidase (GGT) is highly and locally expressed in human breast cancer, a GGT-enzymatically activatable fluorescent probe, gamma-glutamyl hydroxymethyl rhodamine green (gGlu-HMRG), has been developed to detect the human breast cancer area with high performance. In this study, GGT expression and the efficacy of gGlu-HMRG on visualization were investigated in canine mammary gland tumors (MGT). Seventeen non-fixed fresh-frozen MGT specimens and each peritumoral control tissue were utilized. The GGT mRNA levels were highly observed in the tumor specimens compared with the control. GGT immunostaining was mostly observed on the cell membrane and cytosol of the alveolar and duct mammary epithelium of MGT tissues. These signals were strongly positive in several cases while they were mild to not observed in other cases. When gGlu-HMRG solution was dropped to the non-fixed tissue pieces of MGT or control tissues, the fluorescence intensities (FIs) were measured using Maestro in-vivo imaging device. FIs in MGT tissues were significantly higher than each control tissue 20 min after treatment. Based on Youden index method said that the maximum sensitivity and specificity of FI was 82.4% and 82.4%. These findings suggest that GGT is highly expressed in several MGTs in dogs and gGlu-HMRG could visualize at least a part of MGT tissues in dogs. Nevertheless, it should be needed to assess the false-negative areas more carefully in canine than human cases.     

Vertical behavior of juvenile yellowfin tuna Thunnus albacares in the southwestern part of Japan based on archival tagging

The behavior of juvenile yellowfin tuna Thunnus albacares in southwestern Japan was investigated using archival tag data from five fish (fork length 52.5–92 cm, days at liberty 26–280 days) released near the Nansei Islands (24–29°N, 122–130°E). Vertical behavior was classified into three patterns: “shallow” (≥50 % of daytime hours at depth of <50 m), “deep” (≥50 % of daytime hours at ≥100 m), and “intermediate” (other than “shallow” or “deep”). The pooled proportion of the number of days of each behavior was 29, 25 and 46 %, respectively. The proportion of “shallow” behavior increased with fish size. The proportion of time spent near the surface at nighttime increased in the colder season, when the thermal gradient was relatively small. Surface-oriented behavior (fish remained at a depth of <10 m for more than 10 min) occurred mainly during nighttime and between November and January. Dives exceeding 500 m were occasionally observed (0.02 day^?1), and one fish dived to 1230 m. The results of our study show that yellowfin tuna were typically distributed in the mixed layer or upper thermocline where the water temperature was close to the sea surface temperature and that the vertical behavior was variable.     

Developmental states of the collagen content, distribution and architecture in the pectoralis, iliotibialis lateralis and puboischiofemoralis muscles of male Red Cornish x New Hampshire and normal broilers

1. Developmental states of the collagen content, distribution and architecture in the pectoralis (PT), iliotibialis lateralis (ITL) and puboischiofemoralis (PIF) muscles of male Red Cornish x New Hampshire (RN, 80 d, body weight 2.9 kg) and normal (3.1 kg) broilers were evaluated. 2. In PT muscle the total amount of collagen was significantly greater in RN broilers (3.33 mg/g) than in normal ones (1.71 mg/g). This higher collagen content in RN broilers was based mainly on the closer mesh sizes of endomysial honeycomb. The collagen structures in the perimysia also differed between broiler types, when more collagen fibres were observed in RN broilers. 3. ITL muscle contained total collagen of 4.10 to 5.00 mg/g. Types I and III collagens were distributed on the perimysia at higher percentages in RN broilers (31.6%, 37.2%) than normal (15.6%, 30.8%), respectively. The thick bands of tough collagen fibres characteristic of ITL muscle perimysium in cockerels had not yet developed in these broilers. 4. Total collagen was 4.63 to 6.29 mg/g in PIF material with fascia. In PIF muscle the perimysial collagen fibres had not yet attained their full growth but consisted of densely packed fibrils. PIF muscle was characterised by the earlier maturing collagen structure. 5. These results show that a perimysial collagen structure in broilers is still in an undeveloped state. It is supposed that tenderness of broiler meat is attributed mainly to characteristics of the collagen distribution, in which the majority of types I and III collagens is distributed on the closer mesh of endomysial honeycomb.     

Changes to vertical thermoregulatory movements of juvenile bigeye tuna (Thunnus obesus) in the northwestern Pacific Ocean with time of day,seasonal ocean vertical thermal structure,and body size

Vertical movements related to the thermoregulation were investigated in 12 juvenile bigeye tuna (Thunnus obesus) in Japanese waters using archival tag data. Movements changed with time of day, season, and body size. During daytime, bigeye tuna descended to greater depths, presumably to feed in the deep scattering layer (DSL). Thereafter, they repeatedly ascended to shallower layers, suggesting attempts at behavioral thermoregulation, although the beginning of vertical thermoregulatory ascents might reflect a shift in DSL depth. By the end of such movement, the whole‐body heat‐transfer coefficient might decrease because, although the depth and ambient temperature of the upper layers did not change, the body temperature gradually decreased significantly just after ascent for thermoregulation. Seasonal patterns indicated that the vertical thermal structure of the ocean might influence this ascent behavior. For example, from January to May, bigeye tuna made fewer ascents to less shallow waters, suggesting that they respond to increasing depths of the mixed surface layer by reducing energy expenditure during vertical migration. In addition, as body size increased, fewer thermoregulatory ascents were required to maintain body temperature, and fish remained deeper for longer periods. Thus, vertical thermoregulatory movements might change with body size as bigeye tuna develop better endothermic and thermoregulatory abilities. We hypothesize that bigeye might also increase cold tolerance as they grow, possibly due to ontogenetic shifts in cardiac function.     

Grain color development and the inheritance of high anthocyanin blue aleurone and purple pericarp in spring wheat (Triticum aestivum L.)

There is renewed interest in breeding for high anthocyanin content in wheat due to its antioxidant potential. A series of adapted spring wheat lines were developed with blue aleurone or purple pericarp. The development of anthocyanin concentration and color of these selected lines was measured during grain filling for two field seasons at Saskatoon, Canada. In addition, the inheritance of the blue aleurone and purple pericarp was studied. Anthocyanin concentration increased rapidly during grain development and then decreased before maturity. Anthocyanin concentration was highest in PIG03008, a purple pericarp wheat. For mature grain, genotypic variation for anthocyanin concentration was statistically significant while the year and genotype by year interaction were not, facilitating the breeding progress. Blue aleurone was shown to be controlled by a single dominant gene in BC populations whereas purple pericarp appeared to be controlled by two loci with a segregation ratio of 11 purple: 5 white in F₂ populations. The results indicate that breeding high anthocyanin blue or purple wheat is feasible.     

Lesions of bone and bone marrow in myeloid leukosis occurring naturally in adult broiler breeders

Lesions of bone and bone marrow in myeloid leukosis (ML) occurring naturally in adult broiler breeders were investigated pathologically. During gross examination, nodules and protrusions were commonly observed on the surface of the sternum, ribs, vertebrae, and synsacrum. The bone marrow of all the bones of the body was pale in color. Histologically, granulated myelocytes proliferated in the bone marrow of various bones and in the periosteum of the sternum, ribs, vertebrae, and synsacrum. The first proliferation of tumor cells occurred in the bone marrow of epiphysis. The myelocytes invaded through haversian and Volkmann's canals from the bone marrow to periosteal areas. Hematopoiesis was suppressed by marked proliferation of tumor cells in the bone marrow of the whole bone. Atrophy was also seen in the bones, including medullary bones of the chickens suffering from ML. Proliferation of myelocytes was seen in the bone marrow and periosteum of ossified cartilaginous rings of the trachea and larynx. Marked proliferation of myelocytes was seen in the dura mater of spinal cords, and it subsequently depressed the spinal cords. Bone formation with cartilage was seen in the periosteum of the sternum having marked proliferation of myelocytes in the bone marrow and periosteum. Ultrastructurally, tumor cells showed large nuclei and cytoplasm with large round electron-dense lysosomes. The virus particles were rarely detected in the cytoplasm of tumor cells. The polymerase chain reaction test of tumor samples showed positive for subgroup J avian leukosis virus. This study indicates that the myelocytes can invade through the compact bones to the periosteum in the sternum, ribs, vertebrae, synsarcum, and ossified cartilage of trachea and larynx having thinner compact bones. In addition, the periosteal osteogenesis with cartilage in the sternum may be reactive change against the bone atrophy because of the marked proliferation of myelocytes.     

First case of salivary mucocele originating from the minor salivary gland of the soft palate in a dog

We found a case of salivary mucocele that originated in the minor salivary gland (palatine gland) of the soft palate in a dog. At first admission, the soft palate swelled remarkably. Computed tomography (CT) revealed cystic radiolucency inside a large quantity of liquid in the soft palate, and most of the airway was occupied. Marsupialization was performed, but since a recurrence was observed one month later, the salivary mucocele was removed. There has been no report of salivary mucocele arising from the minor salivary gland of the soft palate in dogs. To our knowledge, this case is the first. Complete removal, including minor salivary glands surrounding the lesion, is necessary for treatment of salivary mucocele in dogs.