Genetic and antigenic characterization of bovine viral diarrhea viruses isolated from cattle in Hokkaido,Japan

In our previous study, we genetically analyzed bovine viral diarrhea viruses (BVDVs) isolated from 2000 to 2006 in Japan and reported that subgenotype 1b viruses were predominant. In the present study, 766 BVDVs isolated from 2006 to 2014 in Hokkaido, Japan, were genetically analyzed to understand recent epidemics. Phylogenetic analysis based on nucleotide sequences of the 5′-untranslated region of viral genome revealed that 766 isolates were classified as genotype 1 (BVDV-1; 544 isolates) and genotype 2 (BVDV-2; 222). BVDV-1 isolates were further divided into BVDV-1a (93), 1b (371) and 1c (80) subgenotypes, and all BVDV-2 isolates were grouped into BVDV-2a subgenotype (222). Further comparative analysis was performed with BVDV-1a, 1b and 2a viruses isolated from 2001 to 2014. Phylogenetic analysis based on nucleotide sequences of the viral glycoprotein E2 gene, a major target of neutralizing antibodies, revealed that BVDV-1a, 1b and 2a isolates were further classified into several clusters. Cross-neutralization tests showed that BVDV-1b isolates were antigenically different from BVDV-1a isolates, and almost BVDV-1a, 1b and 2a isolates were antigenically similar among each subgenotype and each E2 cluster. Taken together, BVDV-1b viruses are still predominant, and BVDV-2a viruses have increased recently in Hokkaido, Japan. Field isolates of BVDV-1a, 1b and 2a show genetic diversity on the E2 gene with antigenic conservation among each subgenotype during the last 14 years.     

Relationships between internal ethylene and optical reflectance in ripening ‘Antonovka’ apples grown under sunlit and shaded conditions

The feasibility of non-destructive estimation of internal ethylene concentration (IEC) in apple fruit via fruit reflectance using recently developed approaches and a fiber-optics reflectometer was investigated. The relationships between IEC and fruit reflectance in the 400–800 nm range were studied in stored apple (Malus × domestica Borkh., cv. Antonovka) fruit. A strong correlation between IEC and optical reflectance spectra taken from sunlit surfaces of the fruit was detected whereas reflectance of the shaded fruit surface showed a weak correlation with IEC. The increase of the reflectance in the red occurred along with IEC build-up during ripening resulting a strong (r² > 0.80) correlation. By contrast, reflectance in the blue-green part of the spectrum remained low and was negatively (r² ≈ 0.65) correlated with IEC. These observations are consistent with the phenomenon of degradation of chlorophylls which often occurs in parallel with the retention of carotenoids in ripening apple skin. As a result, IEC showed a significant correlation (r² > 0.69; P < 0.001) with the index based on reflectances in the red and blue-green regions of the spectrum (R₆₇₈ ? R₄₈₀)/R₈₀₀. The effects of strong solar light on the relationships between IEC and fruit reflectance are considered. The possibilities and limitations of a non-destructive reflectance-based assay of IEC in apple fruit are discussed.     

Cold pre-treatment is effective for 1-MCP efficacy in ‘Tsugaru’ apple fruit

1-Methylcyclopropene (1-MCP) treatment maintains apple fruit quality during storage, but its efficacy is dependent on a number of conditions. ‘Tsugaru’ apples are a major early season cultivar in Japan, but because ‘Tsugaru’ fruit produce abundant ethylene, they have a short shelf-life, and efficacy of 1-MCP is not as high with ‘Tsugaru’ as with other cultivars. To improve 1-MCP efficacy, ‘Tsugaru’ fruit were pre-cooled at −1 °C or −3 °C for 24 h before 1-MCP treatment. Ethylene production decreased with the cold treatment, resulting in better storage after 1-MCP treatment. Although ethylene production was low at the end of 24 h of the cold pre-treatment, expression of ACS1, the ethylene receptor genes ERS1, ETR1(a), ETR1b, ETR2 and ETR5, and the cell wall degradation-related gene PG1 all increased with a 24 h cold treatment. It is assumed that these elevated gene expression levels were not caused by ethylene, but more directly by cold stimulus. Thus, a short period of cold stimulus suppresses ethylene production, but induces expression of some genes. 1-MCP treatment was more effective with some initial fruit chilling.     

Carbon and nutrient budgets of the Chagga home garden system in the Kilimanjaro highlands,Tanzania

Sustainable land management of smallholder farms is crucial for ensuring food security in sub-Saharan Africa. However, little is known about the nutrient dynamics of smallholder farming systems at the farm level based on primary data. In this study, carbon (C) and nutrient budgets of the home garden system in the Kilimanjaro highlands, where Andosols predominate, were quantified at the farmer's field. We evaluated (1) the soil C and nutrient flow in the main three land-use blocks (banana (Musa spp.) garden, maize (Zea mays L.) field and grassland) in one representative home garden and (2) the internal flow between farmland and livestock and the external nutrient flow across the inside and outside of the six home gardens. Intensive applications of livestock dung to the banana trees resulted in a positive C budget (7.2 Mg C ha⁻¹ year⁻¹) in the banana garden. Nitrogen loss through the harvesting of feed and crops was almost balanced with the livestock dung application, while nitrogen loss through leaching only accounted for 3% of that applied. Banana productivity has been maintained despite a negative potassium budget (−241 kg K ha⁻¹ year⁻¹), probably owing to the replenished exchangeable potassium from Andosols. In the maize field, the C budget was negative (−1.7 Mg C ha⁻¹ year⁻¹) owing to high organic matter decomposition. Carbon and nutrient budgets in the grassland were all negative. Our results revealed that the village average livestock density (4.4 TLU ha⁻¹: TLU means tropical livestock unit) was sufficient for P, Ca and Mg balance in the home garden, whereas it was not sufficient for N and K balance. Increasing livestock density improved the nutrient balance of the system. However, it is noteworthy that 33%–47% of the feed supplied as C and nutrients was collected from outside the home gardens, suggesting a high reliance on external inflow to fulfil feed demands. In conclusion, intensive livestock dung application to banana cultivation was fundamental for maintaining agricultural productivity to replenish the nutrients lost from the system. At the same time, this system was sustained not only by C and nutrient cycling within the system, but also by transporting resources from the external environment into the system.     

Diverse antimicrobial killing by Enterococcus faecium E 50-52 bacteriocin

An effective bacteriocin was identified and characterized. Lactic acid bacteria were screened against Campylobacter jejuni. One bacteriocin producer, Enterococcus faecium (NRRL B-30746), was studied. The isolate was grown, and the bacteriocin was purified to single-band homogeneity. Biochemical traits indicated that the peptide was a Class IIa bacteriocin, and it was named E 50-52. The bacteriocin had a molecular weight of 3339.7 and an isoelectric point of 8.0. The minimal inhibitory concentrations of E 50-52 against C. jejuni, Yersinia spp., Salmonella spp., Escherichia coli O157:H7, Shigella dysenteriae, Morganella morganii, Staphylococcus spp., and Listeria spp. ranged from 0.025 to 32 microg/mL. In therapeutic broiler trials, oral treatment with E 50-52 reduced both C. jejuni and Salmonella enteritidis by more than 100,000-fold in the ceca, and systemic S. enteritidis was reduced in the liver and spleen. The wide range of antibacterial activity of bacteriocin E 50-52 against pathogens provides a promising alternative to antibiotics.     

Regulation of stony hard peach softening with ACC treatment

Controlling the rate of fruit softening in melting-flesh peaches is a primary goal of the fruit industry. Stony hard (SH) peach varieties lack the ability to synthesize 1-aminocyclopropane-1-carboxylic acid (ACC), the precursor of ethylene, which is required for fruit maturation. SH peaches thus have crisp flesh that remains firm during ripening. In this study, we developed a simple technique to stimulate fruit softening by a single spray application of ACC at a concentration of 10–20 mM, which was sufficient to allow ethylene synthesis and fruit softening. Higher concentrations of ACC increased ethylene production, and made the fruit softer. Ethylene synthesis was limited to the first 2–3 d after ACC treatment, after which fruit ceased softening and retained its remaining firmness. These results indicate that a single application of ACC solution can be used to regulate the process of fruit softening in SH peaches.     

A phylogenomic study of birds reveals their evolutionary history

Deep avian evolutionary relationships have been difficult to resolve as a result of a putative explosive radiation. Our study examined approximately 32 kilobases of aligned nuclear DNA sequences from 19 independent loci for 169 species, representing all major extant groups, and recovered a robust phylogeny from a genome-wide signal supported by multiple analytical methods. We documented well-supported, previously unrecognized interordinal relationships (such as a sister relationship between passerines and parrots) and corroborated previously contentious groupings (such as flamingos and grebes). Our conclusions challenge current classifications and alter our understanding of trait evolution; for example, some diurnal birds evolved from nocturnal ancestors. Our results provide a valuable resource for phylogenetic and comparative studies in birds.     

Spawning induction and seed production of Eastern little tuna,Euthynnus affinis (Cantor, 1849), in the post‐ and pre‐spawning seasons by hormonal treatment in a semi‐closed recirculation system with elevated temperature

We previously established a method for spawning induction in Eastern little tuna (ELT) Euthynnus affinis (Cantor, 1849) by administering a gonadotropin‐releasing hormone analog (GnRHa) during the natural spawning season in Japan (August–October). In order to establish seed production of ELT in the off‐spawning season, we first conducted three spawning induction trials by GnRHa administration from October 2011 to January 2012 using ELT broodstock (2 years old; three females and four males) maintained in a 10‐m³ tank with a semi‐closed recirculation system and static elevated temperature. Average water temperature and daily egg production in three trials lasting 11–15 days were 27.0 ± 0.09°C and 268 173 eggs (Trial 1), 27.0 ± 0.11°C and 277 9098 eggs (Trial 2), and 25.5 ± 0.39°C and 291 113 eggs (Trial 3) respectively. Mean fertilization rate and mean hatching rate were 70.4% and 60.5% (Trial 1), 83.9% and 79.6% (Trial 2), and 62.5% and 57.4% (Trial 3) respectively. We also succeeded in producing ELT larvae in the pre‐spawning season (April–July), although the quantity and quality of larvae produced were inferior to those produced in other calendar months. In trials involving periodic GnRHa administration during the off‐spawning seasons, hatched larvae were obtained in the 10‐m³ tank after six of nine administrations in the 2011–2012 off‐spawning season and in 16 of 19 administrations in the 2012–2013 off‐spawning season. The findings of this study demonstrated that hormonal treatment and thermal control could be used to extend the spawning period in ELT, potentially allowing larval production in the post‐ and pre‐spawning seasons.     

Physicochemical responses of Pythium porphyrae (Oomycota), the causative organism of red rot disease in Porphyra to acidification

Physicochemical responses to acidification in Pythium porphyrae , the causative organism of red rot disease in Porphyra , were investigated. The acid tolerance of P. porphyrae mycelia under pH 4 (acidic condition) condition increased significantly compared with that of the mycelia under pH 8 (condition equivalent to seawater) condition. Free amino acid levels in the mycelia decreased 1.3–8.8-fold under pH 4 condition. However, some free amino acids such as the d -cysteinolic acid-like component, phosphoethanolamine, glutamic acid, aminoadipic acid and methionine increased 2.6–21.7-fold under the same condition. Proton flux on the mycelia exposed to pH 8 increased significantly compared with the mycelia exposed to pH 4. The patterns of proteins present (examined by two-dimensional polyacrylamide gel electrophoresis) differed among the pH conditions. These results suggest that P. porphyrae acquires acid tolerance and is able to adapt to the changing pH conditions. This has significant implications for the use of acidic fungicide treatment for prevention of red rot disease on Porphyra farms.     

Comparison of horse mackerel and tilapia surimi gel based on rheological and <Superscript>1</Superscript>H NMR relaxation properties

ABSTRACT:   Horse mackerel and tilapia surimi were subjected to six different heat and pressure treatments in order to compare gelation characteristics of easy- and difficult-setting gels, that is, temperature dependence, by observing rheological properties and microscopic molecular mobility. The stress–relaxation and proton spin–spin relaxation time (¹H T ₂) of water were measured for all treated gels. Horse mackerel gel demonstrated higher elasticity, large distribution of the stress–relaxation process, and smaller water ¹H T ₂ than tilapia in both heat and pressure treatments. The water ¹H T ₂ was steeply increased in the pressure treatment at around 294 MPa for both fishes. In contrast, the ¹H T ₂ rarely changed in the heat treatment in spite of the considerable change in rheological properties. From the experimental results, it is considered that the gelation of horse mackerel (easy setting) surimi is induced by highly unfolding and re-aggregation of protein, which contributes to the formation of a strong network structure compared with tilapia in both heat and pressure treatments, and that pressure treatments hardly improve the gel strength of tilapia (difficult setting) surimi. The water ¹H T ₂ measurement was used effectively in order to study gelation characteristics of easy- and difficult-setting fish through observing its molecular dynamics.