Rapid and reliable detection of phytoplasma by loop-mediated isothermal amplification targeting a housekeeping gene

Phytoplasmas are plant pathogenic bacteria that infect more than 700 plant species. Because phytoplasma-resistant cultivars are not available for the vast majority of crops, the most common practice to prevent phytoplasma diseases is to remove infected plants. Therefore, developing a rapid, accurate diagnostic method to detect a phytoplasma infection is important. Here, we developed a phytoplasma detection assay based on loop-mediated isothermal amplification (LAMP) by targeting the groEL gene and 16S rDNA. We designed 19 primer sets for the LAMP assay and evaluated their amplification efficiency, sensitivity, and spectra to select the most suitable primer sets to detect Candidatus Phytoplasma asteris. As a result, DNA was efficiently amplified by one of the primer sets targeting the groEL gene, and LAMP assay sensitivity with this primer set was 10-fold higher than that of the polymerase chain reaction. Moreover, the groEL gene was successfully amplified from several strains of Ca. Phytoplasma asteris by this primer set, indicating that the groEL gene can be used as a LAMP assay target gene for a broad range of phytoplasma strains. Additionally, a simple DNA extraction method that omits the homogenizing and phenol extraction steps was combined with the LAMP assay to develop a simple, rapid, and convenient diagnostic method for detecting phytoplasma.     

Dietary Protein Hydrolysates from Animal By-Products: Digestibility and Enzymatic Activity for Dourado Salminus brasiliensis

Fishmeal (FM) is a high-cost and scarce feedstuff. Notwithstanding, the formulation and processing of diets for carnivorous fish strongly rely on FM as a protein source, given its palatability and biological value. Animal by-product hydrolysates are high-quality feedstuff and can substitute FM in aquafeeds. This study evaluated the digestibility of hydrolysates from tilapia residue (TR), tuna head (TH), swine liver (SL), and poultry liver (PL), and the profile of digestive enzymes in juvenile dourado, Salminus brasiliensis, a carnivorous Characin, fed diets containing graded levels of the hydrolysates. Inclusion of hydrolysates in diet formulations lowered the pH of feed but did not alter feed intake by fish. Higher apparent digestibility coefficients (ADCs) of diets were recorded for fish fed diets containing TR and SL, and lower ADCs were recorded for fish fed diets containing TH. Protease and lipase activities in the animal’s stomach were higher, especially for those fed with diets containing SL. Amylase activity was higher in pyloric caeca, whereas in the intestines, the higher activity was seen in fish fed control and TR diets. Animal by-product hydrolysates were highly digestible for dourado, and the enzymatic profile of fish was influenced by the nutrient contents of diets.     

First report of ‘<Emphasis Type="Italic">Candidatus</Emphasis> Phytoplasma malaysianum’ associated with <Emphasis Type="Italic">Elaeocarpus</Emphasis> yellows of <Emphasis Type="Italic">Elaeocarpus zollingeri</Emphasis>

“Elaeocarpus yellows” (ELY) is a widely reported phytoplasma disease of Elaeocarpus zollingeri trees in Japan. The phytoplasma associated with ELY (ELY phytoplasma) had not been identified at the species level because its 16S rRNA sequence had yet to be reported. Here, we report the results of a sequence analysis based on 16S rRNA and secA gene sequences, which showed that the ELY phytoplasma is related to ‘Candidatus Phytoplasma malaysianum’. To our knowledge, this is the first report showing the occurrence of ‘Ca. P. malaysianum’ outside Malaysia and the infection of E. zollingeri by the phytoplasma.     

Growth variability of Pacific saury Cololabis saira larvae under contrasting environments across the Kuroshio axis: survival potential of minority versus majority

Growth variability was examined for Pacific saury Cololabis saira larvae under contrasting environments across the Kuroshio axis, based on samples collected during the winter spawning season in 2013 and 2014. The growth rate index (residual of the otolith marginal 3‐day mean increment width from the linear regression on knob length) of larvae was compared among three areas: the inshore side of the Kuroshio axis, the Kuroshio axis, and the offshore side of the Kuroshio axis in relation to sea surface temperature (SST), salinity (SSS) and chlorophyll‐a (CHL) concentration. The larvae were more densely distributed in the Kuroshio axis and offshore areas of higher temperature and salinity and lower chlorophyll‐a concentration than in the inshore areas of lower temperature and salinity and higher chlorophyll‐a concentration. No marked differences in the growth rate index were found among the three areas, even though the larvae in the inshore areas showed slightly higher growth rates in 2013. Despite the broad ranges of environmental factors, no clear relationship between the growth rate index and any environmental factor was detected. The survival potential of Pacific saury larvae was considered to be at least comparable under contrasting environments across the Kuroshio axis. Such a geographical homogeneity is concluded to be attributable to compensable effects of physical and biological factors. We hypothesize that the minority under physically‐unfavorable but biologically‐favorable conditions on the inshore side of the Kuroshio axis could survive equally well as the majority under physically‐favorable but biologically‐unfavorable conditions around the Kuroshio axis and on the offshore side of the Kuroshio axis.     

A fructose‐based extender protects Colossoma macropomum spermatozoa against chilling injuries

Our study assessed the efficiency of a formulated new extender in maintaining viability and morphological integrity of Colossoma macropomum spermatozoa under chilling storage. Semen was diluted in the test extender and BTS? (Beltsville Thawing Solution) and exposed to a short‐term storage at 4.6 ± 0.6°C for 96 hr. Both extenders were able to maintain 17% ± 8% motile spermatozoa by the end of experiment. Sperm dilution in test extender did not affect the morphologically normal cells (61% ± 6%) up to 48 hr of chilling, being higher than in BTS? (50% ± 6%) (p < 0.05). After 96 hr, samples kept in the test extender had 50% of normal spermatozoa, whereas those kept in BTS? presented only 38% of normal cells. Chilling storage increased the incidence of cells with strongly coiled flagella in BTS^?. Our study is the first to evaluate in detail the spermatozoa morphology as indicative of C. macropomum semen viability. The new extender was able to protect the spermatozoa against increase in coiled flagellum injuries.     

Measurement of Indoor Sulfur Dioxide Emission from Coal–Biomass Briquettes

The coal–biomass briquette is an alternative fuel that replaces low-grade coal. It is expected that the use of coal–biomass briquettes will effectively reduce the indoor concentration of sulfur dioxide (SO₂) emitted during raw coal combustion. In this study, indoor concentrations of SO₂ emitted from combustion of either coal–biomass briquettes or low-grade coal in households in Nanchuan, which is in the rural area of Chongqing City, China, were measured. The acidic gases were collected using passive samplers, and the samplers were taken to the laboratory for extraction and measurement. The SO₂ concentrations resulting from the use of coal–biomass briquettes were clearly reduced compared with those resulting from the use of low-grade coal. The concentrations of SO₂ to which the inhabitants (housewives) of the houses were exposed were also measured, and were found to be less with coal–biomass briquettes. Although with coal–biomass briquettes exposure concentrations still exceeded the World Health Organization guideline for SO₂ (125 μ g m^?3), indoor air pollution and adverse effects on human health caused by SO₂ could be decreased by substituting coal–biomass briquettes for low-grade coal as a domestic fuel.     

First report of bacterial leaf blight on cosmos (Cosmos bipinnatus Cav.) caused by Pseudomonas cichorii in Japan

Since 2006, a commercial grower in Japan has noted a leaf blight symptom on potted cosmos plants grown in a field. In August 2012, a Pseudomonas-like bacterium was isolated from the symptomatic leaves and found to cause the same symptom on cosmos seedlings after inoculation. On the basis of bacteriological and phylogenetic analyses, the causative bacterium was identified as Pseudomonas cichorii. This is the first report of bacterial leaf blight on cosmos caused by P. cichorii in Japan.     

Simulated Acid Rain Leaching Characteristics of Acid Soil Amended with Bio-Briquette Combustion Ash

In Chongqing City, the rapid growth of the economy has accompanied an increase of sulfur dioxide emissions from coal combustion, bringing about an expansion of acid rain affected areas and acidification of soil. Recently, we reported that coal-biomass briquettes so-called bio-briquettes (BB), which are produced from pulverized raw coal, biomass, and a sulfur fixation agent (Ca (OH)₂) under 3 to 5 tons cm^?2 pressure, have high sulfur-fixation efficiency. The BB ash contains nutritive substances such as Ca and Mg, and has a large acid-neutralizing capacity. Thus, in order to improve the acid soil in the Chongqing area, we analyzed the chemical composition of the original acid soil and the ash-amended soil, and investigated their leaching characteristics under simulated acid rain (SAR). It was found that plants and crops in Chongqing area would be injured if the present acid rain continues. We carried out a SAR leaching experiment and studeid the potential toxic effects of leachate from soil containing added ash. The results indicated that the contents of most toxic elements, with the sole exception of chromium, were below the environment standard for irrigation water. Because the BB ash was highly alkaline, the leaching aluminum (Al) species would be hydroxide rather than free Al³⁺ ion.