Identification of enzyme genes in the liver of the Bleeker’s squid Loligo bleekeri by expressed sequence tag analysis

Expressed sequence tag (EST) analyses were performed with the aim of identifying enzyme genes in the liver of the Bleeker’s squid Loligo bleekeri. Of the 768 ESTs identified and sequenced, 669 were grouped into 324 clusters. Of these clusters, 123 comprising 245 ESTs were found to be homologous to genes reported to date. Among these, 43 clusters were annotated as enzymes according to the Enzyme Commission (EC) numbering system. Two EC groups, oxidoreductases and hydrolases, possessed a large number of ESTs. A cluster homologous to the glutathione peroxidase, an enzyme in the oxidoreductase group, contained 16 ESTs, which accounted for 2.4% of the total ESTs sequenced. There are three serine proteases, three cathepsins, two triacylgricerol lipases, and two chitinases among the clusters homologous to the enzymes in the hydrolase group. Since the squid liver functions in the digestive process, these enzymes would be involved in food digestion. Our data provide information on the various types of enzymes expressed in the squid liver and may provide a useful basis for further characterization of these enzymes.     

Effects of high-K+, Na+-deficient solution on contractility of the smooth muscles of the bovine trachea

A high-K+, Na+-deficient (I-154 K+) solution induced contraction followed by gradual relaxation of the smooth muscles of the bovine trachea, while hyperosmotic addition of 65 mM KCl induced a large sustained contraction. Exposure of the muscle to the I-154 K+ solution induced an increase in the ratio of cellular water content and a sustained increase in oxidized flavoprotein fluorescence or reduced pyridine nucleotide fluorescence. The I-154 K+ solution also induced a sustained increase in [Ca2+]i level. Decreases in developed tension and increases in cellular water content were both prevented by the addition of sucrose or NaCl but not pyruvate. Substitution of KI for KCl in the I-154 K+ solution produced a greater inhibition of contraction, while substitution with K-propionate produced no inhibition of contraction. Moreover, decreases in developed tension and increases in cellular water content were both prevented by addition of 100 microM 4,4'-diisothiocyanatostilbene-2,2'-disulphonic acid (DIDS), but not 10 microM bumetanide or 1 mM acetazolamide. In conclusion, I-154 K+ solution induced-relaxation in the bovine trachea may be due to swelling of smooth muscle cells and the mechanism of swelling is probably involved in DIDS-sensitive anion movement.     

Experimental infection of pigs with different doses of the African swine fever virus Armenia 07 strain by intramuscular injection and direct contact

We experimentally infected pigs with the African swine fever virus (ASFV) Armenia 07 strain (genotype II) to analyze the effect of different dose injections on clinical manifestations, virus-shedding patterns, histopathology, and transmission dynamics by direct contact. Each three pigs and four pigs were injected intramuscularly with 0.1 fifty percent hemadsorbing doses (HAD₅₀)/ml, 10¹ HAD₅₀/ml and 10⁶ HAD₅₀/ml of ASFV Armenia 07 strain, respectively. Each two of three pigs injected with 0.1 HAD₅₀/ml and 10¹ HAD₅₀/ml died by 10 days post inoculation. All pigs had a gross lesion of splenomegaly. Perigastric and renal lymph nodes were enlarged and resembled blood clots in nine of ten pigs. It was revealed that 0.1 HAD₅₀/ml of this ASFV was sufficient to infect healthy pigs by intramuscular injection and caused sub-acute lethal disease. For the transmission study, two 8-week-old pigs were injected intramuscularly with 10³ HAD₅₀/ml of the same virus. Each of the experimentally inoculated pigs was co-housed with two 8-week-old naive pigs. All contact pigs exhibited clinical manifestations at 6 or 7 days after the experimentally inoculated pigs developed pyrexia. These findings suggest that this strain may spread slowly within a herd. Histologically, lymph nodes resembled blood clots were formed by severe blood absorption and followed hemorrhage result of disruption of the lymphoid sinus filling with absorbed red blood cells. The severity of the gross and histological lesions depended on duration after infection, regardless of the difference of injection doses in this study.     

Practicable group testing method to evaluate weight/weight GMO content in maize grains

Because of the increasing use of maize hybrids with genetically modified (GM) stacked events, the established and commonly used bulk sample methods for PCR quantification of GM maize in non-GM maize are prone to overestimate the GM organism (GMO) content, compared to the actual weight/weight percentage of GM maize in the grain sample. As an alternative method, we designed and assessed a group testing strategy in which the GMO content is statistically evaluated based on qualitative analyses of multiple small pools, consisting of 20 maize kernels each. This approach enables the GMO content evaluation on a weight/weight basis, irrespective of the presence of stacked-event kernels. To enhance the method's user-friendliness in routine application, we devised an easy-to-use PCR-based qualitative analytical method comprising a sample preparation step in which 20 maize kernels are ground in a lysis buffer and a subsequent PCR assay in which the lysate is directly used as a DNA template. This method was validated in a multilaboratory collaborative trial.     

Three-dimensional underwater shape measurement of tuna longline using ultrasonic positioning system and ORBCOMM buoy

ABSTRACT:   The installation depth of the hook in longline fishing gear has previously been measured with micro depth loggers. Research that assumes the catenary shape of longline fishing gear by a simulation based on these data has been done. However, it was not known whether the branch line was tangled with the main line or flowed with the current. In this research, an ultrasonic positioning system generally used to investigate the underwater behavior of marine organisms, and a buoy with a communications satellite, have been used, and the 3-dimensional underwater shape of tuna longline fishing gear was measured. It was possible to monitor changes in fishing gear in real time. The possibility of high precision measurement was suggested with future technical improvement.     

Dataset of CarboEastAsia and uncertainties in the CO2 budget evaluation caused by different data processing

The datasets of net ecosystem CO₂ exchange (NEE) were acquired from 21 forests, 3 grasslands, and 3 croplands in the eastern part of Asia based on the eddy covariance measurements of the international joint program, CarboEastAsia. The program was conducted by three networks in Asia, ChinaFLUX, JapanFlux, and KoFlux, to quantify, synthesize, and understand the carbon budget of the eastern part of Asia. An intercomparison was conducted for NEE estimated by three gap-filling procedures adopted by ChinaFLUX, JapanFlux, and KoFlux to test the range of uncertainty in the estimation of NEE. The overall comparison indicated good agreement among the procedures in the seasonal patterns of NEE, although a bias was observed in dormant seasons depending on the different criteria of data screening. Based on the gap-filled datasets, the magnitude and seasonality of the carbon budget were compared among various biome types, phenology, and stress conditions throughout Asia. The annual values of gross primary production and ecosystem respiration were almost proportional to the annual air temperature. Forest management, including clear-cutting, plantation, and artificial drainage, was significant and obviously affected the annual carbon uptake within the forests. Agricultural management resulted in notable seasonal patterns in the crop sites. The dataset obtained from a variety of biome types would be an essential source of knowledge for ecosystem science as well as a valuable validation dataset for modeling and remote sensing to upscale the carbon budget estimations in Asia.     

Agricultural insect hybridization and implications for pest management

Biological invasions, the expansion of agricultural frontiers, and climate change favor encounters of divergent lineages of animals and plants, increasing the likelihood of hybridization. However, hybridization of insect species and its consequences for agroecosystems have not received sufficient attention. Gene exchange between distinct and distant genetic pools can improve the survival and reproduction of insect pests, and threaten beneficial insects in disturbed agricultural environments. Hybridization may be the underlying explanation for the recurrent pest outbreaks and control failures in putative hybrid zones, as suspected for bollworm, corn borer, whiteflies, and stink bugs. Reliable predictions of the types of changes that can be expected in pest insect genomes and fitness, and of their impacts on the fate of species and populations remain elusive. Typical steps in pest management, such as insect identification, pest monitoring, and control are likely affected by gene flow and adaptive introgression mediated by hybridization, and we do not have ways to respond to or mitigate the problem. To address the adverse effects of farming intensification and global trade, we must ensure that current integrated pest management programs incorporate up‐to‐date monitoring and diagnostic tools. The rapid identification of hybrids, quantification of levels of introgression, and in‐depth knowledge of what genes have been transferred may help to explain and predict insect population outbreaks and control failures in the future. © 2019 Society of Chemical Industry