Determination of the genetic structure of remnant Morus boninensis Koidz. trees to establish a conservation program on the Bonin Islands, Japan

Background  

Morus boninensis, is an endemic plant of the Bonin (Ogasawara) Islands of Japan and is categorized as "critically endangered" in the Japanese red data book. However, little information is available about its ecological, evolutionary and genetic status, despite the urgent need for guidelines for the conservation of the species. Therefore, we adopted Moritz's MU concept, based on the species' current genetic structure, to define management units and to select mother tree candidates for seed orchards.     

Effect of simulated acid fog on membrane-bound calcium (mCa) in fir (Abies firma) and cedar (Cryptomeria japonica) mesophyll cells

The decline of virgin fir (Abies firma) forest at Mt. Oyama has been reported. Related field observations suggest that high acidity fog is linked with its decline. However, cedar (Cryptomeria japonica) in the same area shows no symptoms of decline. For assessing effects of acid fog on membrane-bound calcium (mCa) of the leaf mesophyll cells, 9-year-old seedlings of fir (Abies firma) and 8-year-old seedlings of cedar (Cryptomeria japonica) were exposed twice a week to simulated acid fog (SAF at pH 3 with pH 5 as control) for 2 h per day in a chamber during May–December 2007 (except August). Current and 1-year-old needles were collected from seedlings and analyzed at 1-month intervals. For current-year needles of fir, mCa levels in cells exposed to SAF at pH 3 were significantly lower than in cells exposed to pH 5, especially during September 2007–March 2008. In contrast, it is noteworthy that mCa levels of cedar were maintained as virtually constant irrespective of SAF acidity, indicating that fir is more sensitive to acid fog than is cedar. Based on these results, mCa loss by acid fog might also be caused in the declining virgin fir forest at Mt. Oyama.     

Effects of Brief Summer or Winter Fasting on the Muscle Quality of Cultured Pacific Bluefin Tuna (Thunnus orientalis)

The effects of fasting on the quality of the dorsal and ventral ordinary muscles from cultured Pacific bluefin tuna (Thunnus orientalis) during chilled storage were investigated. Tuna were subjected to fasting for 2 days in the summer or 6 days in the winter prior to harvesting. The breaking strength of the dorsal ordinary muscle sampled in the summer increased until 24 h and then decreased. There were no significant differences in the lipid and glycogen content of the ordinary muscle after 9 h of storage between the controls and either fasting group. The pH of the ordinary muscle subjected to summer and winter fasting was higher than in the controls after 24–48 h of storage. However, the relationship between the pH and glycogen content was unclear. The metmyoglobin content during chilled storage was lower in the ordinary muscles from either fasting group than in the controls. In conclusion, fasting for 6 days in the winter improved the color stability of the ordinary muscle without a decline in its lipid content.     

Dormancy Characteristics of Malus ‘Snowdrift’ and Effects of Spraying 6-benzyladenine on Its Germination and Flowering in Long-day Treatment

[Objective] The dormancy characteristics of Malus ‘Snowdrift’ in long-day treatment were studied, and 6-BA was used to break the dormancy, with the aim to achieve the purpose of flowering in autumn. [Method] The new shoots of ‘Snowdrift’ in long-day treatment were conducted with hydroponics to investigate their dormancy time. And cytokinin 6-benzyladenine(6-BA)was used to treat the dormant shoots, to investigate the budding, flowering and flower bud differentiation. [Result] The shoots in long-day treatment entered endodormancy after August 7~(th) and the shoots in the natural daylight entered endodormancy before July 18~(th). In long-day treatment, 116 buds, 198 flowers were observed after 6-BA spraying. [Conclusion] The optimum concentration of 6-BA was 300 mg/L. And the flower bud differentiation of ‘Snowdrift’ in long-day treatment was faster than those in natural daylight after 6-BA spraying.     

Investigation of lactic acid bacterial strains for meat fermentation and the product's antioxidant and angiotensin‐I‐converting‐enzyme inhibitory activities

In the lactic acid bacteria (LAB) strains screened from our LAB collection, Lactobacillus (L.) sakei strain no. 23 and L. curvatus strain no. 28 degraded meat protein and tolerated salt and nitrite in vitro. Fermented sausages inoculated strains no. 23 and no. 28 showed not only favorable increases in viable LAB counts and reduced pH, but also the degradation of meat protein. The sausages fermented with these strains showed significantly higher antioxidant activity than those without LAB or fermented by each LAB type strain. Angiotensin‐I‐converting‐enzyme (ACE) inhibitory activity was also significantly higher in the sausages fermented with strain no. 23 than in those fermented with the type strain. Higher ACE inhibitory activity was also observed in the sausages fermented with strain no. 28, but did not differ significantly from those with the type strain. An analysis of the proteolysis and degradation products formed by each LAB in sausages suggested that those bioactivities yielded fermentation products such as peptides. Therefore, LAB starters that can adequately ferment meat, such as strains no. 23 and no. 28, should contribute to the production of bioactive compounds in meat products.     

Frequent use of colistin-based drug treatment to eliminate extended-spectrum beta-lactamase-producing <Emphasis Type="Italic">Escherichia coli</Emphasis> in backyard chicken farms in Thai Binh Province,Vietnam

Reports of livestock infections with extended-spectrum beta-lactamase-producing Escherichia coli (ESBL-E) are increasing. Based on interviews conducted over a 6-month period, we found that veterinarians in the Vietnamese province of Thai Binh prefer to prescribe colistin-based drugs (CBD) in chicken farms. We aimed to clarify whether CBD use selects for strains of colistin-resistant ESBL-E. With the cooperation of seven local households, we detected ESBL-E in chickens’ feces after treating chickens with CBD. Phylogenetic groupings and the presence of CTX-M/AmpC genes were determined, and the multi-antibiotic susceptibility of isolates was analyzed. Our results showed that ESBL-E presented in seven chickens’ feces from two households. Seventy-two percent of ESBL-E isolates harbored CTX-M9 and the phylogenetic group A; the colistin minimum inhibitory concentration (MIC) of all isolated ESBL-E ranged from 0.064 to 1 μg mL^?1. Moreover, ESBL-E isolates were used to experimentally select for colistin resistance, and the effect of commercial CBD on ESBL-E was investigated. The results showed that an ESBL-E strain with a colistin MIC of 4 μg mL^?1 was able to grow in media with CBD. Although CBD treatment was effective, in vitro experiments demonstrated that ESBL-E can easily acquire colistin resistance. Therefore, restrictions on colistin use are necessary to prevent the emergence of colistin-resistant bacteria.     

Early metaphase II oocytes treated with dibutyryl cyclic adenosine monophosphate provide suitable recipient cytoplasm for the production of miniature pig somatic cell nuclear transfer embryos

We investigated the effects of in vitro maturation duration and treatment with dibutyryl cyclic adenosine monophosphate (dbcAMP) on the blind enucleation efficiency and developmental competence of miniature pig somatic cell nuclear transfer (SCNT) embryos. Oocytes were cultured for 22 h in NCSU-23 medium with or without 1 mM dbcAMP and then additionally cultured in dbcAMP-free NCSU-23 for 14, 18, or 22 h. Regardless of dbcAMP treatment, the rate of nuclear maturation reached a plateau at 36 and 40 h. However, mitochondrial distribution, a marker for cytoplasmic maturation, differed between the dbcAMP-untreated oocytes at 36 h and dbcAMP-treated oocytes at 40 h. The metaphase II chromosomes were adjacent to the first polar body in 68.8% and 63.5% of the dbcAMP-untreated oocytes at 36 h and dbcAMP-treated oocytes at 40 h, respectively. Furthermore, the blind enucleation efficiency by removing a small volume of cytoplasm was significantly higher in the dbcAMP-untreated oocytes at 36 h (82.9%) and dbcAMP-treated oocytes at 40 h (89.9%) than other groups. The rate of blastocyst formation was highest in the dbcAMP-treated oocytes at 40 h. Hence, this study demonstrated that dbcAMP-treated early metaphase II oocytes are suitable for the production of miniature pig SCNT embryos.     

Mapping of QTLs controlling epicotyl length in adzuki bean (Vigna angularis)

Epicotyl length (ECL) of adzuki bean (Vigna angularis) affects the efficiency of mechanized weeding and harvest. The present study investigated the genetic factors controlling ECL. An F₂ population derived from a cross between the breeding line ‘Tokei1121’ (T1121, long epicotyls) and the cultivar ‘Erimo167’ (common epicotyls) was phenotyped for ECL and genotyped using simple sequence repeats (SSRs) and single-nucleotide polymorphism (SNP) markers. A molecular linkage map was generated and fifty-two segregating markers, including 27 SSRs and 25 SNPs, were located on seven linkage groups (LGs) at a LOD threshold value of 3.0. Four quantitative trait loci (QTLs) for ECL, with LOD scores of 4.0, 3.4, 4.8 and 6.4, were identified on LGs 2, 4, 7 and 10, respectively; together, these four QTLs accounted for 49.3% of the phenotypic variance. The segregation patterns observed in F₅ residual heterozygous lines at qECL10 revealed that a single recessive gene derived from T1121 contributed to the longer ECL phenotype. Using five insertion and deletion markers, this gene was fine mapped to a ~255 kb region near the end of LG10. These findings will facilitate marker-assisted selection for breeding in the adzuki bean and contribute to an understanding of the mechanisms associated with epicotyl elongation.     

Reproductive cycle observation of the Okinawa rail (Gallirallus okinawae) in the wild

The captive breeding program of the Okinawa rail started in 2008. For successful captive breeding, information related to reproduction, such as age at sexual maturity, testicular cycles and ovulatory cycles, is essential to predict when reproduction is possible and when certain reproductive behaviors are most likely to occur. We made gross and histological observations of the reproductive organs of Okinawa rails to gain understanding of sexual maturity, the testicular cycle and the ovulatory cycle. We found that the weight of the testis was smallest in December and largest in March. Changes in the diameter of the seminiferous tubule showed the same pattern. Mature sperm were observed from March to June. The heaviest ovary was observed in April. A single peak of reproduction, from March to April, was observed in males and females. Our observations suggested that the Okinawa rail is a seasonal breeder. Establishing suitable breeding pairs will be critical to ensure success of the Okinawa rail captive breeding program. Our results suggested that pairing must be started before March. If supportive breeding is used, semen should be collected from March to June and artificial insemination conducted in April.     

Prevalence of equine herpesvirus type 1 strains of neuropathogenic genotype in a major breeding area of Japan

A single non-synonymous nucleotide substitution of guanine (G) for adenine (A) at position 2254 in the viral DNA polymerase gene (encoded by open reading frame [ORF] 30) of equine herpesvirus type 1 (EHV-1) has been significantly associated with neuropathogenic potential in strains of this virus. To estimate the prevalence of EHV-1 strains with the neuropathogenic genotype (ORF30 G(2254)) in the Hidaka district--a major horse breeding area in Japan--we analyzed the ORF30 genomic region in cases of EHV-1 infection in this area during the years 2001-2010. Of the 113 cases analyzed, 3 (2.7%) were induced by ORF30 G(2254) strains. This prevalence is lower than those observed in the U.S.A. (10.8-19.4%), Argentina (7.4%), France (24%), and Germany (10.6%).