Quantitative loop-mediated isothermal amplification method for the detection of <Emphasis Type="Italic">Vibrio nigripulchritudo</Emphasis> in shrimp

Vibrio nigripulchritudo is considered one of the major pathogens threatening shrimp aquaculture. In this study, we developed a novel and highly specific quantitative loop-mediated isothermal amplification (Q-LAMP) assay. A set of four specific primers were designed targeting the V. nigripulchritudo intergenic spacer region. The reaction time and temperature were optimized for 60 min at 63°C. Quantitative analysis was then performed by measuring the turbidity of the reaction solution using a real-time turbidimeter, allowing for quantification of the initial DNA concentration with a sensitivity of 10² copy numbers equivalent to 2.3 colony forming units/ml or 0.3 fg/μl. The LAMP assay was able to specifically detect two representative strains of V. nigripulchritudo, whereas other Vibrio and non-Vibrio species were not amplified. A standard curve was generated for V. nigripulchritudo by plotting the threshold time (T _t) versus the log of bacterial number. A high correlation coefficient (R ² = 0.9749) was observed for the Q-LAMP reaction. In conclusion, Q-LAMP assay is a sensitive, rapid, and simple tool that can be used for the detection and quantification of V. nigripulchritudo in shrimp, thereby facilitating surveillance of vibriosis infection.     

Synthesis and Structure–Activity Relationships of Miticidal 4,5-Dihydropyrazole-5-thiones

A series of novel 4,5-dihydropyrazole-5-thiones (DHPs) was synthesised by treating the corresponding dihydropyrazolones with ‘Lawesson’s reagent and evaluated for miticidal activity against two-spotted spider mites (Tetranychus urticae Koch). Of these, 3-(4-chlorophenyl)-4,4-dimethyl-1-phenyl-4,5-dihydropyrazole-5-thione, 3-(4-chlorophenyl)-4-ethyl-4-methyl-1-phenyl-4,5-dihydropyrazole-5-thione, 3-(4-chlorophenyl)-1-phenyl-4,5-dihydropyrazole-5-thione-4-spirocyclopentane and 4,4-dimethyl-1-phenyl-3-(4-trifluoromethyl-phenyl)-4,5-dihydropyrazole-5-thione were highly active (pEC₅₀>4·0) and were more effective than the miticide dicofol (pEC₅₀=3·879), which has traditionally been used for the control of phytophagous mites. Structure–activity relationship (SAR) studies were performed on each position of the pyrazole ring of DHPs. The results indicated that the unsubstituted phenyl, 4-substituted phenyl and thioxo groups on the 1-, 3- and 5-positions of DHPs respectively were required for activity. Quantitative SAR studies using physicochemical parameters of substituents and the capacity factor k′ as a hydrophobicity index suggested that: (a) the activities of all types of DHPs examined were mainly dominated by hydrophobicity, (b) the bulkiness of 4-substituents of the 3-phenyl ring favoured the activity and (c) the log k′ optimum for all DHPs was 1·675, equivalent to a log P_ow value of c. 5·0.     

Contrasting seasonal leaf habits of canopy trees between tropical dry-deciduous and evergreen forests in Thailand

We compared differences in leaf properties, leaf gas exchange and photochemical properties between drought-deciduous and evergreen trees in tropical dry forests, where soil nutrients differed but rainfall was similar. Three canopy trees (Shorea siamensis Miq., Xylia xylocarpa (Roxb.) W. Theob. and Vitex peduncularis Wall. ex Schauer) in a drought-deciduous forest and a canopy tree (Hopea ferrea Lanessan) in an evergreen forest were selected. Soil nutrient availability is lower in the evergreen forest than in the deciduous forest. Compared with the evergreen tree, the deciduous trees had shorter leaf life spans, lower leaf masses per area, higher leaf mass-based nitrogen (N) contents, higher leaf mass-based photosynthetic rates (mass-based P(n)), higher leaf N-based P(n), higher daily maximum stomatal conductance (g(s)) and wider conduits in wood xylem. Mass-based P(n) decreased from the wet to the dry season for all species. Following onset of the dry season, daily maximum g(s) and sensitivity of g(s) to leaf-to-air vapor pressure deficit remained relatively unchanged in the deciduous trees, whereas both properties decreased in the evergreen tree during the dry season. Photochemical capacity and non-photochemical quenching (NPQ) of photosystem II (PSII) also remained relatively unchanged in the deciduous trees even after the onset of the dry season. In contrast, photochemical capacity decreased and NPQ increased in the evergreen tree during the dry season, indicating that the leaves coped with prolonged drought by down-regulating PSII. Thus, the drought-avoidant deciduous species were characterized by high N allocation for leaf carbon assimilation, high water use and photoinhibition avoidance, whereas the drought-tolerant evergreen was characterized by low N allocation for leaf carbon assimilation, conservative water use and photoinhibition tolerance.     

Decline of <Emphasis Type="Italic">Pinus thunbergii</Emphasis> Parlat. stands due to excess soil moisture caused by a buried andosol layer at a coastal sand site in Hokkaido,northern Japan

To demonstrate the effect of excess soil moisture on the decline of a coastal Pinus thunbergii stand in Oshamanbe, southwestern Hokkaido, Japan, soil moisture content was monitored for 4 years. The saturated hydraulic conductivities (K _S) of different soil types (coastal sand, supplied topsoil, and buried concreted andosol) and the distribution of the buried concreted andosol layer were investigated. We also examined needle length to verify the real-time response of P. thunbergii to excess soil moisture. Soil moisture content at the heavily damaged site was more heterogeneous than that at the slightly damaged site, and a sensor near the ground always reported a higher soil moisture content at the heavily damaged site than at the slightly damaged site. The buried concreted andosol layer was always found at the heavily damaged site. The K _S of the andosol layer was 10⁻⁵, suggesting that this layer is less permeable to water, leading to excess soil moisture at this site. P. thunbergii needles from the heavily damaged site were shorter than those from the slightly damaged site, possibly because of water stress. Together with other symptoms observed at the study sites, i.e., crown dieback and intense lateral growth, this information leads us to conclude that the decline of P. thunbergii stands at the heavily damaged site in Oshamanbe was caused by excess soil moisture due to the less permeable buried concreted andosol layer.     

Use of soil color meter for aqueous iron and ammonium measurements

Abstract

In this paper, we proposed a new approach for on-site colorimetric analysis of ferrous ions (Fe²⁺) and ammonium-nitrogen (NH₄ ⁺-N) using a soil color meter as an alternative method to conventional spectrophotometry. The soil color meter we used can express solution color numerically on the basis of L*a*b* color space. After coloring of water by the 1, 10 phenanthroline method and the Indophenol blue method, the color of solution was measured by the soil color meter. A linear relationship between Fe²⁺ and a* or b* values, and systematic change of NH₄ ⁺-N with L* value, enable us to make a calibration curve. The Fe²⁺ and NH₄ ⁺-N concentrations in groundwater samples (Fe²⁺: 0.3–1.3 mg L^?1; NH₄ ⁺-N: 0.02–0.62 mg L^?1) determined by the proposed method agreed well with those determined by conventional spectrophotometry with the difference being ± 0.05 mg L^?1 and ± 0.02 mg L^?1, respectively. Since a similar apparatus is widely used in the soil science field, this technique would facilitate field surveys.     

The Use of Bacillus Licheniformis HA1 To Accelerate Composting of Organic Wastes

The effect of Bacillus licheniformis HA1 cell density on the acceleration of organic waste composting was tested in a bench-scale composting system utilizing a process limit temperature of 60°C. Variables measured during composting were CO₂ evolution rate, conversion of substrate carbon and pH. When an initial cell density of 2.0×10⁴ cfu/g-dry solid was used, the strain HA1 increased in number and prevented the decrease in pH during the early stage of composting. This resulted in enhanced populations of other thermophiles and increased the rate of organic matter decomposition. By contrast, no effect was observed at a lower cell density of HA1. It was found that the minimum cell density of HA1 to accelerate organic decomposition was around 10⁴-10⁵ cfu/g dry solid of raw material.     

A next-generation-sequencing approach to diagnose viral diseases of greenhouse ranunculus

Journal of General Plant Pathology - Viruses that infect Ranunculus asiaticus L., a globally important greenhouse ornamental, have consistently caused serious damage in Japan. We coupled...     

中国对拉美农产品出口的影响因素分析及前景展望

利用引力模型,应用1995-2004年的贸易数据,考察了中国对拉美主要国家农产品出口的影响因素。计算结果显示,中国农产品供给能力、进口国GDP以及距离因素是中国对拉美农产品出口的重要影响因素。进一步的分析表明。中国对拉美主要贸易伙伴的农产品出口属于“贸易活跃”状况。其中乌拉圭的农产品市场已经得到较深入的开拓。而委内瑞拉、阿根延和厄瓜多尔等3个国家的出口潜力还需要进一步挖掘。     

Predominant expression of growth hormone secretagogue receptor in the caudal lobe of chicken pituitary and parallel developmental increase in expression of pituitary GH and proventriculus ghrelin mRNA

To examine the involvement of ghrelin in growth hormone (GH) synthesis in the chicken pituitary, the regional distribution of GH secretagogue receptor (GHS‐R)/ghrelin receptor was investigated. Quantitative real‐time polymerase chain reaction (Q‐PCR) analysis revealed that the expression levels of GHS‐R and GH mRNA in the caudal lobe were about fourfold and sevenfold higher in the cephalic lobe of 7 day‐old chickens, respectively. Immunohistochemical analysis showed that GHS‐R immunoreactivity was more abundant in the caudal lobe than in the cephalic lobe, as was the case for GH immunoreactivity. By Q‐PCR, parallel increases were observed in the expression levels of ghrelin mRNA in the proventriculus and GH mRNA in the pituitary from embryonic day 17 to day 7 after hatching, whereas no significant change was found in the expression levels of GHS‐R mRNA in the pituitary during this period. These results suggest that proventriculus‐derived ghrelin may participate in pituitary GH synthesis by acting on its receptor during late embryonic development and the early post‐hatching period in chickens.