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.     

In situ detection of cellular and abnormal isoforms of prion protein in brains of cattle with bovine spongiform encephalopathy and sheep with scrapie by use of a histoblot technique.

To detect prion protein, brains from 5 cattle naturally affected with bovine spongiform encephalopathy (BSE) and 3 sheep naturally affected with scrapie were examined and compared with brains of normal cattle and sheep using a histoblot technique. The technique enabled the in situ distinctive detection of the cellular (PrP(C)) and abnormal (PrP(Sc)) isoforms of the prion protein. In BSE- or scrapie-affected brains, the Prp(C) signal decreased, especially in those areas where the PrP(Sc) signal was detected.     

Food production and the energy crisis

The principal raw material of modern U.S. agriculture is fossil fuel, whereas the labor input is relatively small (about 9 hours per crop acre). As agriculture is dependent upon fossil energy, crop production costs will also soar when fuel costs increase two- to fivefold. A return of 2.8 kcal of corn per 1 kcal of fuel input may then be uneconomical. Green revolution agriculture also uses high energy crop production technology, especially with respect to fertilizers and pesticides. While one may not doubt the sincerity of the U.S. effort to share its agricultural technology so that the rest of the world can live and eat as it does, one must be realistic about the resources available to accomplish this mission. In the United States we are currently using an equivalent of 80 gallons of gasoline to produce an acre of corn. With fuel shortages and high prices to come, we wonder if many developing nations will be able to afford the technology of U.S. agriculture. Problems have already occurred with green revolution crops, particularly problems related to pests (57). More critical problems are expected when there is a world energy crisis. A careful assessment should be made of the benefits, costs, and risks of high energy-demand green revolution agriculture in order to be certain that this program will not aggravate the already serious world food situation (58). To reduce energy inputs, green revolution and U.S. agriculture might employ such alternatives as rotations and green manures to reduce the high energy demand of chemical fertilizers and pesticides. U.S. agriculture might also reduce energy expenditures by substituting some manpower currently displaced by mechanization. While no one knows for certain what changes will have to be made, we can be sure that when conventional energy resources become scarce and expensive, the impact on agriculture as an industry and a way of life will be significant. This analysis is but a preliminary investigation of a significant agricultural problem that deserves careful attention and greater study before the energy situation becomes more critical.     

Fatal suppurative meningoencephalitis caused by Klebsiella pneumoniae in two calves

One calf died (No. 1) and another was euthanized following astasia (No. 2). Histopathological examination revealed suppurative meningoencephalitis in these calves. Klebsiella pneumoniae antigens were detected in lesions. Thymocytes were decreased in the thymus cortex in both cases. 16S rRNA gene sequencing of the No. 1 isolate and bacterial extracts from formalin fixed paraffin embedded sections of No. 2 revealed that both samples were K. pneumoniae. The No. 1 isolate showed multidrug resistance against penicillin antibiotics, fosfomycin, streptomycin, macrolide antibiotics, tetracycline antibiotics, and clindamycin. Immunosuppression is a significant septicemic K. pneumoniae infection risk factor. Our study provides new aspects regarding K. pneumoniae infections in cattle, bacterial meningoencephalitis differentiation, and K. pneumoniae and bacterial meningoencephalitis treatments.     

Molecular analysis of β-proteobacterial ammonia oxidizer populations in surface layers of a submerged paddy soil microcosm

The structure of the β-proteobacterial autotrophic ammonia-oxidizing bacterial (AOB) communities in a microcosm of submerged paddy soil was determined by denaturing gradient gel electrophoresis (DGGE) analysis of 16S rRNA gene fragments amplified using AOB-selective primers. Shift in the community composition was observed 4 weeks after submergence. The communities from the surface layers (0–1, 2–3 mm) of the soil microcosm were different from those of the subsurface layers (6–9, > 15 mm) and DGGE bands specific to each layer were detected. The majority of the retrieved sequences were Nitrosospira-like, whereas no Nitrosomonas-like sequences were obtained. The 16S rDNA primer set also amplified sequences that were not related to the known Nitrosospira-Nitrosomonas group, although they showed a close relationship with other groups of β-proteobacteria. The results suggest that Nitrosospira-like populations are dominant AOB populations in the submerged paddy soil, and that the oxic layer of submerged paddy soil harbours the specific AOB.     

Characterization of the Na+ delivery from roots to shoots in rice under saline stress: Excessive salt enhances apoplastic transport in rice plants

Sodium transport to the shoots of rice (Oryza sativa L.) plants grown under salt stress conditions was characterized. The rate of Na⁺ transport to shoots increased exponentially depending on the increase in the NaCl concentration of the rooting solution, however, the rates were independent of the plant transpiration. Excessive NaCl in the rooting solution was found to enhance apoplastic transport in rice plants by using the apoplastic dye, 3-hydroxy-5,8,10-pyrene trisulfonic acid (PTS) and Fluostain I (also known as Calcoflour White M2R New). The results suggest that excessive Na⁺ in the rooting solution enlarges the apoplastic pathways, which facilitates Na⁺ intrusion into the xylem vessels, resulting in an excessive accumulation of Na⁺ in rice shoots.     

Influence of intergeneric/interspecies mitochondrial injection; parthenogenetic development of bovine oocytes after injection of mitochondria derived from somatic cells

Interspecies/intergeneric mitochondrial heteroplasmy can occur in interspecies/intergeneric hybrid embryos or following nuclear transfer. In the present study, intergeneric buffalo (Bubalus bubalis) mitochondria (WB-mt) or interspecies murine (Mus spretus) mitochondria (M-mt) were injected into bovine (Bos taurus) oocytes, and the subsequent embryonic development was characterized. Fibroblast mitochondria (WB-mt or M-mt) were microinjected into in vitro matured bovine oocytes followed by oocyte activation by a combination of electrical stimulation and 6-dimethylaminopurine treatment. After seven days of culture, embryo development was evaluated. The copy number of specific mtDNA populations (introduced and native mtDNA) from heteroplasmic oocytes was estimated using real-time PCR. The results illustrated that oocytes injected with either WB-mt or M-mt can develop to the blastocyst stage (20.6% and 19.6%). Cleavage division rates and development to the morula stage in oocytes injected with WB-mt were lower (76.2% and 45.9%, respectively) in comparison with uninjected oocytes (89.2% and 59.1%, respectively) (P<0.05). However, no differences were found in comparing M-mt injected oocytes and controls (P>0.05). An increase in bovine mtDNA copy number was observed at the expanded blastocyst stage of injected embryos (P<0.01), while the number of injected mtDNA was stable throughout development. This study demonstrates that interspecies/intergeneric mitochondrial injected bovine oocytes have the ability to develop to the blastocyst stage after parthenogenetic activation and that injected mtDNA was neither selectively destroyed nor enhanced through development. Moreover, injected intergeneric mitochondria had a demonstrated influence on bovine parthenogenetic development and mtDNA replication.     

Local and Systemic Induced Resistance to the Root-Knot Nematode in Tomato by DL-beta-Amino-n-Butyric Acid

ABSTRACT Chemical inducers of pathogenesis-related proteins and plant resistance were applied to tomato plants, with the aim of inducing resistance to the root-knot nematode Meloidogyne javanica. Relative to control plants, foliar spray and soil-drenching with dl-beta-amino-n-butyric acid (BABA) reduced root-galling 7 days after inoculation, as well as the number of eggs 30 days after inoculation. Other chemicals (alpha- and gamma-amino-n-butyric acid, jasmonic acid, methyl jasmonate, and salicylic acid) were either phytotoxic to tomato plants or did not improve control of root-knot nematodes. Fewer second-stage juveniles invaded BABA-treated tomato roots, and root-galling indices were lower than in control tomato plants. Resistance phenomena in seedlings lasted at least 5 days after spraying with BABA. Nematodes invading the roots of BABA-treated seedlings induced small, vacuolate giant cells. Postinfection treatment of tomato plants with BABA inhibited nematode development. It is speculated that after BABA application tomato roots become less attractive to root-knot nematodes, physically harder to invade, or some substance(s) inhibiting nematode or nematode feeding-site development is produced in roots.