Involvement of two different types of Verticillium dahliae in lettuce wilt in Ibaraki Prefecture, Japan

Although wilt diseases of various plants by Verticillium dahliae occur worldwide, Verticillium wilt of lettuce is relatively new and rare. In 2009, this disease appeared suddenly in commercial fields in Ibaraki Prefecture, Japan. To investigate the cause of the disease, we characterized each lettuce isolate for pathogenicity, mating types, and genetic characteristics and found two types of V. dahliae. A population that included at least two types of V. dahliae was the putative infection source of the disease. Effective management systems against V. dahliae isolates of different races and having different host ranges are necessary.     

Use of microsatellite DNA and amplified fragment length polymorphism for Cherry salmon (Oncorhynchus masou) complex identification

Formosa landlocked salmon (Oncorhynchus masou formosanus), an endemic, critically endangered subspecies of Cherry salmon (Oncorhynchus masou) complex, is only found in Taiwan. Because the eyed eggs and ungutted carcasses of Pacific salmons (genus Oncorhynchus) are imported for aquaculture and food to Taiwan from overseas every year, the requirement for preventing illegal trade or accidental commercial imports to avoid unwanted fish from contaminating the gene pool of Formosa landlocked salmon and infect them with diseases is critical for the conservation of Formosa landlocked salmon. Traditional morphology‐based species identification is impossible for salmon eggs and larvae that lack clearly defined morphological features. In the present study, the genetic differences among four subspecies (Oncorhynchus masou ishikawae, O. masou subsp., Oncorhynchus masou masou and O. masou formosanus) of Cherry salmon complex were determined from microsatellite DNA and amplified fragment length polymorphism analyses. We successfully generated a genetic marker to aid traditional taxonomy and investigate the integrity of the current taxonomic status among members of Cherry salmon complex. Use of molecular markers, in combination with traditional morphological identification, is a promising tool for identifying four closely related subspecies of Cherry salmon complex.     

Nephrocalcinosis formation by soy isoflavones in male rats

Isoflavones (IFs), found in the form of both aglycones and glucosides in soybean foods, induce weak estrogenic activities. Although IFs have a number of health benefits, it was previously reported that IFs cause nephrocalcinosis (NC) in the kidney of male Fischer 344 (F344) rats. The present study aims to elucidate the safety of IFs by focusing on IF-induced NC formation in rats. Fermented soybean extract (FSE) containing 420 mg/g isoflavone aglycones was orally administered to male F344 and Sprague-Dawley (SD) rats for 28 days. FSE induced NC formation in the kidney of F344 rats, but not in SD rats. However, absorption of IFs did not differ between F344 and SD rats. NC formation and its severity of FSE were histologically compared with those of soybean extract (SE) containing 518 mg/g isoflavone glucosides in F344 rats. There were no differences in the number of NC formations and the extent of calcium deposit between FSE and SE groups. To examine the dose effect of FSE on NC formation, doses of 20, 140, or 1000 mg/kg FSE were administered to F344 rats for 90 days. NC formation was observed in the 140 and 1000 mg/kg groups. These results indicated that a high dose of oral administration of IFs induced NC formation depending on the strain of rat.     

Estimating the three-dimensional structure of canopy foliage based on the light measurements in a Betula ermanii stand

In order to more accurately evaluate the functional activity of forest stands by canopy production and evapotranspiration, we improved the methods for field measurements and statistical modeling to estimate foliage configuration (spatial distribution of leaves) while simultaneously reconstructing the three-dimensional photosynthetically active photon flux density (PPFD) distribution (PPFD pattern) in a forest canopy. By using a sensor (photodiode) array, a PPFD pattern was observed in summer 2002 under the canopy in an even-aged, pure stand of Japanese mountain birch Betula ermanii Cham. (17-years old) in Hokkaido, northern Japan. A Markov chain Monte Carlo (MCMC) sampling technique is developed such that a set of foliage configurations generated by the model referred to as the Gibbs foliage canopy (GFC) approximates the field-measured PPFD pattern. The posterior distribution of the foliage configurations is generated by the parallel tempering MCMC of eight independent series of foliage configurations. The GFC model generated the posterior distribution of the LAI estimates (mean 4.56) that appeared to be appropriate in comparison to other LAI estimates of the B. ermanii stand based on the indirect and nondestructive methods by LAI-2000 (LAI = 3.43) and litterfall traps (LAI = 5.56) because they could be under- and overestimated, respectively. Our evaluations of the canopy production and evapotranspiration rates suggest that the relationship between LAI and canopy functions was not very simple because it depended on the nonlinear functional forms of the leaf responses of photosynthesis and transpiration to PPFD. The current study demonstrates an application of MCMC techniques that can generate a set of possible structures of unobserved/unobservable objects based on the high-resolution dataset obtained by some indirect (or remote-sensing) methods.     

Clarification of relationship between single-nucleotide polymorphism panels of Shiga toxin-producing Escherichia coli O157:H7/H- strains

Eighty strains of enterohemorrhagic Escherichia coli O157:H7/H- were analyzed by three single-nucleotide polymorphism (SNP) panels using whole-genome sequencing data. The partial concordance of SNP types among the different SNP panels was observed on minimum spanning trees reconstructed with SNP data. As for lineage I/II strains, some of the clade 7 strains belonged to one unique SNP type as determined by three panels, suggesting that clade 7 should be divided into at least two genotypes, namely, the unique type and the rest. In addition, clade 8 contained two unique genotypes, which was consistent with the previous prediction. Similarly, for lineage II, clade 12 should be divided into three genotype strains. In contrast, many strains of several clades belonging to lineage I were clustered into the same node on each minimum spanning tree upon testing with the three SNP panels. Previous studies reported that lineage I diverged more recently than lineages I/II and II. Such low diversity in lineage I in this study may have arisen because this lineage has not accumulated SNPs because of its relatively recent divergence. Based on the concordance observed in this study, some of the previously published O157 genotype distribution data were successfully interpreted to clarify the clade distribution, which was well supported by previous literature.     

Non-contact respiratory measurement in a horse in standing position using millimeter-wave array radar

This study aimed to apply radar technology to a large quadruped animal. We first developed a non-contact respiration measurement system using millimeter-wave array radar for a horse in standing position. Specifically, we measured the respiration of a stationary domestic horse in stables. Simultaneously, we measured the respiration rate using infrared thermography and developed a method for analyzing the radar information while verifying the rate of agreement. Our results suggested that the radar technology detected breathing and accurately measured the respiration of a horse, despite variation in the breathing frequency. To the best of our knowledge, this is the first study to apply a non-contact respiration measurement system using millimeter-wave array radar has been applied to large animals in an upright position, thereby demonstrating its potential application in animal husbandry and welfare.     

Molecular epidemiological analysis of bat rabies viruses in Brazil

A molecular epidemiological analysis was performed in 19 rabies viruses (RVs) isolated from haematophagous, frugivorous and insectivorous bats, in Sao Paulo, Brazil. The authors carried out RT-PCR for amplification of the RV nucleoprotein (N) gene, and determined 1,335 nucleotide sequences of N gene by direct sequencing method. Phylogenetic analysis, which was based on the N gene of Brazilian RV isolates identified presently and previously, revealed that RVs isolated from bats were genetically divided into four lineages had a tendency to depend on the host bat species. The first lineage consisted mainly of haematophagous bat (Desmodus rotundus) isolates, including frugivorous bat (Artibeus spp.) isolates. Other three lineages consisted of insectivorous bat isolates; mainly Eptesicus spp., Molossus spp. and Nyctinomops spp. isolates, respectively. These results indicate a possibility of that there are bat species-specific RV variants in Brazil.     

Oncolytic reovirus synergizes with chemotherapeutic agents to promote cell death in canine mammary gland tumor

The oncolytic effects of reovirus in various cancers have been proven in many clinical trials in human medicine. Oncolytic virotherapy using reovirus for canine cancers is being developed in our laboratory. The objective of this study was to examine the synergistic anti-cancer effects of a combination of reovirus and low doses of various chemotherapeutic agents on mammary gland tumors (MGTs) in dogs. The first part of this study demonstrated the efficacy of reovirus in canine MGTs in vitro and in vivo. Reovirus alone exerted significant cell death by means of caspase-dependent apoptosis in canine MGT cell lines. A single injection of reovirus impeded growth of canine MGT tumors in xenografted mice, but was insufficient to induce complete tumor regression. The second part of this study highlighted the anti-tumor effects of reovirus in combination with low doses of paclitaxel, carboplatin, gemcitabine, or toceranib. Enhanced synergistic activity was observed in the MGT cell line treated concomitantly with reovirus and in all the chemotherapeutic agents except toceranib. In addition, combining reovirus with paclitaxel or gemcitabine at half dosage of half maximal inhibitory concentration (IC₅₀) enhanced cytotoxicity by activating caspase 3. Our data suggest that the combination of reovirus and low dose chemotherapeutic agents provides an attractive option in canine cancer therapy.     

P and N deficiency change the relative abundance and function of rhizosphere microorganisms during cluster root development of white lupin (Lupinus albus L.)

We studied microbe-plant interactions of white lupin, a cluster root-forming plant, under low P and N conditions to examine increased nutrient acquisition by plants either by a shift to a more specialized microbial community or changes in microbial enzyme production. White lupin plants were grown in rhizoboxes filled with either P- or N-deficient soil; fertilized soil was used as control. After cultivation of plants in a glasshouse for 41 d, plant growth (shoot and roots) and P and N accumulation in shoots were measured. Microbial functions were analyzed by P- and N-cycling enzymes. The microbial community structure was estimated by fingerprinting (denaturing gradient gel electrophoresis) and sequencing techniques. P deficiency induced the released citrate and acid phosphomonoesterases from cluster roots and stimulated the production of microbe-derived alkaline phosphomonoesterase in the rhizosphere. P deficiency decreased microbial diversity in the cluster root rhizosphere. Increased relative abundance of Burkholderiales in the rhizosphere of P deficient plants might be responsible for the degradation of different organic P fractions such as phytates. N deficiency induced an increase of the number of nodules and P concentration in shoot as well as roots of white lupin. We clarified that high release of citrate from cluster roots might be the preferred mechanisms to meet the P demand of nodulated plants under N deficiency. In addition, the high abundance of Rhizobiales and Rhodospirillales in the rhizosphere of cluster roots showed that the importance of N-fixing microorganisms under N deficiency. The contribution of rhizosphere microorganisms due to similar activities of N-cycling enzymes under the two different N treatments is less important for N nutrition of plants. Further understanding of the regulation of cluster roots under N-deficiency will provide new information on the interactions between P and N nutrition.