Genetic and enzymatic analyses of metalloprotease (aureolysin) from Staphylococcus aureus isolated from domestic animals.

The gene (aur) encoding the metalloprotease (aureolysin) of Staphylococcus aureus from domestic animals was analyzed by polymerase chain reaction (PCR), PCR-restriction fragment length polymorphism (PCR-RFLP) and sequencing. The aur gene was detected in all 74 isolates from cows, pigs and chickens by PCR amplification and was classified into types I and II by PCR-RFLP patterns. The type II aur gene was contained in 36 (94.7%) of 38 protease-positive isolates as judged by skim milk agar plate culture, while type I was contained in 28 (77.8%) of 36 protease-negative isolates. The deduced amino acid sequences of aureolysins from type I and II isolates were almost identical with those of the published data. Subsequently, the two aureolysins were purified from the culture supernatants of type I and II isolates. The molecular weights of purified type I and II aureolysins were both estimated at 34kDa by SDS-polyacrylamide gel electrophoresis. These aureolysins had maximum proteolytic activity at 30-50 degrees C and pH 7.0-8.0. Their activity was inhibited by metal- and zinc-specific inhibitors, such as EDTA, EGTA and 1,10-phenanthroline. Specific activity (activity/protein) of type II aureolysin was two times higher than that of type I. These results indicated that the aur gene is highly conserved with two allelic forms (types I and II) among bovine, porcine and avian isolates of S. aureus.     

Influence of precipitations and sunshine hours on yield of paddy field grown wheat (Triticum aestivum L.) in Northern Kyushu,Japan

ABSTRACT

Relationships between wheat (Triticum æstivum L.) yield components and growing season weather conditions (e.g. temperature, precipitation, and sunshine duration) were investigated for a rice-wheat rotation paddy system situated in Northern Kyushu, Japan over a 17-year period (2000–2016). A 1.0 mm increase in precipitation from 21 to 32 days after sowing decreased wheat yield by about 27 kg ha^?1 at early seedling stage with one to two leaves (Zadoks (Z) growth stage Z1.1–1.2). Number of tillers and spikes showed significant negative correlations to the amount of precipitation during the period. An hour increase in mean daily sunshine hours from 94 to 111 days after sowing resulted in an 328 kg ha^?1 rise in grain yield at stem elongation stage with the first and second node detection (Z3.1–3.2). The grain number per square meter also showed strong positive correlations to sunshine hours during the same period. However, this positive effect of sunshine was negated when precipitation exceeded about 30 mm during the early seedling stage. Consequently, precipitations at early seedling stage and sunshine hours at stem elongation stage mostly determined wheat yield in Northern Kyushu paddy field. Increasing precipitations during the early seedling stage highlights the pressing need for effective paddy field drainage management from wheat sowing through harvest.

Abbreviations: CP: Chikushi Plains; FARC: Fukuoka prefectural Agriculture and Forestry Research Center; SARC: Saga prefectural Agricultural Research Center; DAS: days after sowing     

Bacterial Pleuritis with Thickened Mesothelial Hyperplasia in a Young Beagle Dog

A five-month-old male beagle dog suddenly became moribund. Bloody fluid accumulated in the thoracic and abdominal cavities, and soft yellow flecks were floating in the thoracic fluid. The mediastinum and pericardium became dark reddish with villous thickening. Other parietal and pulmonary pleurae were rough, and the organs adhered to each other. Histologically, most mediastinal pleura formed papillary projections covered by a single layer of mesothelial cells. Many macrophages and neutrophils infiltrated the submesothelial connective tissue. At the mediastinum adjacent to the pericardium, cuboidal mesothelial cells proliferated solidly and formed a thick surface stratum. The flecks consisted of gram-negative filamentous or small bacillary (coccoid) bacteria. In the right posterior lobe of the lung, neutrophilic infiltration and a large encapsulated abscess including a bacterial colony were present. We diagnosed this case as “bacterial pleuritis with thickened mesothelial hyperplasia”. The cause of the pleuritis might be a chronic pleural infection spread via the lung abscess.     

Insecticidal and neuroexciting actions of DDT analogs

The neuroexciting activity of DDT and its analogs to produce repetitive responses on the nerve cord of Periplaneta americana was determined using the extracellular electrode method. The convulsive activity on P. americana and the insecticidal effect on Callosobruchus chinensis were also examined. It was found that the convulsive and insecticidal activities increase almost proportionally with increase in the neuroexciting activity within a set of p,p′-substituted DDT analogs. The intimate connections among these biological effects suggest that symptoms such as convulsion and death caused by DDT analogs are closely related with their neuroexcitory effect and there is a common mode of action in spite of differences in insect species.     

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.