Manipulation of fish germ cell: visualization, cryopreservation and transplantation

Germ-cell transplantation has many applications in biology and animal husbandry, including investigating the complex processes of germ-cell development and differentiation, producing transgenic animals by genetically modifying germline cells, and creating broodstock systems in which a target species can be produced from a surrogate parent. The germ-cell transplantation technique was initially established in chickens using primordial germ cells (PGCs), and was subsequently extended to mice using spermatogonial stem cells. Recently, we developed the first germ-cell transplantation system in lower vertebrates using fish PGCs and spermatogonia. During mammalian germ-cell transplantation, donor spermatogonial stem cells are introduced into the seminiferous tubules of the recipient testes. By contrast, in the fish germ-cell transplantation system, donor cells are microinjected into the peritoneal cavities of newly hatched embryos; this allows the donor germ cells to migrate towards, and subsequently colonize, the recipient genital ridges. The recipient embryos have immature immune systems, so the donor germ cells can survive and even differentiate into mature gametes in their allogeneic gonads, ultimately leading to the production of normal offspring. In addition, implanted spermatogonia can successfully differentiate into sperm and eggs, respectively, in male and female recipients. The results of transplantation studies in fish are improving our understanding of the development of germ-cell systems during vertebrate evolution.     

Prevalence of Shiga toxin-producing Escherichia coli in Yezo sika deer (Cervus nippon yesoensis) in the Tokachi sub-prefecture of Hokkaido,Japan

In food hygiene, the surveillance of foodborne pathogens in wild animals is indispensable because we cannot control hygienic status of them. Yezo sika deer (Cervus nippon yesoensis), which are found only on the island of Hokkaido, Japan, are the most common game animal in the country. In this study, we analyzed the incidence of Shiga toxin-producing Escherichia coli (STEC) in Yezo sika deer hunted in the Tokachi sub-prefecture, which is one of the densest zones for the sub-species. Real-time polymerase chain reaction testing detected STEC in 18.3% of fecal samples (59/323) collected from deer hunted between 2016 and 2017, whereas no Shigella and Salmonella markers were detected. No correlation was found between STEC detection from fecal samples and characteristics of carcasses, such as hunting area, age, and fascioliasis. From 59 STEC-positive fecal samples, we isolated 37 STEC strains, including 34 O- and H-genotyped strains, in which 16 different serogroups were detected. Genetic analysis revealed that our isolates included various stx gene types (stx1⁺/stx2⁻, stx1⁺/stx2⁺, and stx1⁻/stx2⁺) and carried eae. This study demonstrated that STEC strains with various features colonized the Yezo sika deer, similar to other subspecies of sika deer. We conclude that continuous surveillance activity is important to monitor the suitability of game animals as a food source and to assess the validity of the food safety management system for game meat production.     

Effects of crown length on indole acetic acid (IAA) amounts in cambial region tissues in lower and upper trunks of sugi cultivars (Cryptomeria japonica) in September

Indole acetic acid (IAA) was believed to be an important regulator in xylem formation in conifers. However, few studies have been done on the endogenous amount of IAA in cambial region tissues in sugi (Cryptomeria japonica) trees. In this study, we report the IAA amounts in the lower and upper trunk of mature sugi cultivars (Kumotoshi, Yaichi and Obiaka) in September, and the effects of growth traits on the IAA amounts. The effect of height position (lower or upper trunk) on IAA amounts was found to be larger than that of genetic variation (Kumotoshi, Yaichi or Obiaka) by two-way ANOVA. There was no significant interaction effect (genetic variation × height position). The IAA amounts of all trees varied from 3 to 42 ng/cm² at the lower trunk and 9 to 47 ng/cm² at the upper trunk. Crown length and distance from crown base had significant positive and negative effects on IAA amounts in the trunk of sugi cultivars, respectively. Distance from crown base had a larger effect on IAA amounts than crown length. In Yaichi, taller trees had larger IAA amounts at both the lower and upper trunk.     

Experimental evidence on prevention of infection by the ectoparasitic protozoans Ichthyobodo salmonis and Trichodina truttae in juvenile chum salmon using ultraviolet disinfection of rearing water

In northern Japan, juvenile chum salmon Oncorhynchus keta (Walbaum) are released from hatcheries to enhance the fishery resource. Infections with ectoparasitic protozoans, particularly the flagellate Ichthyobodo salmonis and the ciliate Trichodina truttae, occasionally cause severe mortality among hatchery‐reared juveniles. This study examined the susceptibility of the two parasites to wide‐ranging UV irradiation (experiment 1) and then investigated whether UV disinfection of the rearing water using a commercial device was useful for preventing infections among juveniles in a small‐scale rearing system over a 28‐day period (experiment 2). In experiment 1, parasite mortality reached 100% with UV irradiation doses of ≥9.60 × 10⁵ μW s/cm² for I. salmonis and ≥8.40 × 10⁵ μW s/cm² for T. truttae. In experiment 2, disinfection of the rearing water at a UV irradiation dose of 2.2 × 10⁶ μW s/cm² succeeded in complete prevention of both parasites in the juvenile salmon. These results elucidate the minimum dose of UV irradiation for inactivation of I. salmonis and T. truttae, and demonstrate the usefulness of water disinfection using a commercial UV irradiation device to prevent infections by these parasites in hatchery‐reared juvenile chum salmon.     

Reduction of formaldehyde emission from plywood using composite resin composed of resorcinol–formaldehyde and urea-modified scallop shell nanoparticles

More than 200,000 tons of scallop shells are disposed annually alone in Japan. Nanoparticles derived from scallop shells have the potential to adsorb gaseous formaldehyde; therefore, such discarded shells have now been tested as additive filler in plywood adhesive by mixing high specific surface area, urea-modified shell nanoparticles with a resorcinol–formaldehyde resin. With this procedure, it was found that the emission of formaldehyde from the resulting plywood could be substantially reduced. The urea-modified scallop shell nanoparticles were prepared by two different methods: (1) a dry method where the shells were treated by planetary ball-grinding under ambient conditions—a completely dried powder was obtained after addition of the surface-modifying urea solution; (2) a moist method by treating dry ground shell particles in a wet grinding process with the urea solution, followed by centrifugation to obtain a paste. The specific surface area of the nanoparticles obtained by both treatments was 42 ± 3 m²/g. Measurement of the subsequent formaldehyde emission showed that the addition of the modified scallop shell nanoparticles substantially reduced the formaldehyde emission from plywood. The reduction of the specific mass uptake of urea depends on the nanoparticles which especially was the case when resins containing nanoparticles processed by the moist method were used.     

Laparoscopic cryptorchidectomy in standing bulls

Laparoscopic cryptorchidectomy without insufflation was applied in 10 standing bulls aged 3 to 15 months. Nine bulls were preoperatively pointed out intra-abdominal testes by computed tomography. Preoperative fasting for a minimum of 24 hr provided laparoscopic visualization of intra-abdominal area from the kidney to the inguinal region. Surgical procedure was interrupted by intra-abdominal fat and testis size. It took 0.6 to 1.5 hr in 4 animals weighing 98 to 139 kg, 0.8 to 2.8 hr in 4 animals weighing 170 to 187 kg, and 3 and 4 hr in 2 animals weighing 244 and 300 kg to complete the cryptorchidectomy. In conclusion, standing gasless laparoscopic cryptorchidectomy seems to be most suitable for bulls weighing from 100 to 180 kg.     

Changes of hip joint congruity after triple pelvic osteotomy in the dog with hip dysplasia

Changes in hip joint congruity was evaluated in dogs with hip dysplasia before and after triple pelvic osteotomy by computed tomography examination in the standing position. Lateral center edge angle significantly increased, and center distance (CD) significantly decreased after surgery compared to the values before surgery, respectively. There was an inverse proportion between the postoperative period and the change in the ratio of CD. These results suggested that joint laxity was improved with time after surgery, providing evidence of the clinical usefulness of this surgery.     

Death losses due to stillbirth, neonatal death and diseases in cloned cattle derived from somatic cell nuclear transfer and their progeny: a result of nationwide survey in Japan

To obtain the data concerning death losses due to stillbirth, neonatal death and diseases in cloned cattle derived from somatic cell nuclear transfer (SCNT) and their progeny produced by Japanese institutions, a nationwide survey was carried out in July-August, 2006. As a result, lifetime data concerning 482 SCNT cattle (97.5% of cattle produced in the country at that time) and 202 progeny of SCNT cattle were accumulated and the death loss of these cattle was analyzed. Although 1/3 of delivered SCNT calves died during the perinatal period due to stillbirth and neonatal death, incidence of death loss due to diseases in SCNT cattle surviving more than 200 days after birth seems to be the same as these in conventionally bred cattle. In contrast, progeny of SCNT cattle showed the same level in death loss as observed in conventionally bred cattle throughout their lifetime. These results suggest that robust health would be expected in SCNT cattle surviving to adulthood and their progeny.