Production of Functional Gametes from Cryopreserved Primordial Germ Cells of the Japanese Quail

The Japanese quail (Coturnix japonica) is a valuable bird as both an experimental animal, for a wide range of scientific disciplines, and an agricultural animal, for the production of eggs and meat. Cryopreservation of PGCs would be a feasible strategy for the conservation of both male and female fertility cells in Japanese quail. However, the effects of freeze-thaw treatment on viability, migration ability and germline transmission ability of quail PGCs still remain unclear. In the present study, male and female PGCs were isolated from the blood of 2-day-old embryos, which were cooled by slow freezing and then cryopreserved at –196 C for 77–185 days, respectively. The average recovery rate of PGCs after freeze-thawing was 47.0%. The viability of PGCs in the frozen group was significantly lower than that of the control group (P<0.05) (85.5% vs. 95.1%). Both fresh and Frozen-thawed PGCs that were intravascularly transplanted into recipient embryos migrated toward and were incorporated into recipient gonads, although the number of PGCs settled in the gonads was 48.5% lower in the frozen group than in the unfrozen control group (P<0.05). Genetic cross analysis revealed that one female and two male recipients produced live progeny derived from the frozen-thawed PGCs. The frequency of donor-derived offspring was slightly lower than that of unfrozen controls, but the difference was not significant (4.0 vs. 14.0%). These results revealed that freeze-thaw treatment causes a decrease in viability, migration ability and germline transmission ability of PGCs in quail.     

Inoculation with N2-generating denitrifier strains mitigates N2O emission from agricultural soil fertilized with poultry manure

Pelleted poultry manure is recommended for use with agricultural soil as a replacement for chemical fertilizers; however, application of the manure stimulates nitrous oxide (N₂O) emission from the soil through denitrification. To mitigate the N₂O emission caused by application of pelleted poultry manure, soil microcosms were set up; each microcosm was inoculated with one of the following N₂-generating denitrifier strains previously been isolated from paddy soil: Azoarcus, Dyella, Dechloromonas, Niastella, and Burkholderia. When pelleted poultry manure was incubated on its own, N₂O was produced by denitrification. In contrast, N₂O emission was significantly lowered when the manure was inoculated with most of the N₂-generating strains. In soil microcosms, N₂O was emitted during incubation after application of the pelleted manure, while N₂O flux was significantly lowered when the soil was inoculated with Azoarcus sp. KS11B, Niastella sp. KS31B, or Burkholderia sp. TSO47-3 on the 12th day of incubation. In addition, when pelleted manure was inoculated with the strains prior to application in the soil microcosms, the level of N₂O emission was significantly lowered to ca. 40–60 % that from the non-inoculated control. Our study provides the prototype of a technique that uses microbial technology to mitigate N₂O emission from agricultural soil fertilized with pelleted poultry manure.