Ultra highly sensitive method for detecting Flavobacterium psychrophilum using high‐gradient immunomagnetic separation with a polymerase chain reaction

We attempted to develop an ultrahigh sensitive method for detecting Flavobacterium psychrophilum using high‐gradient immunomagnetic separation (HGIMS) with a polymerase chain reaction (PCR). HGIMS is a magnetic separation method in which the magnetic force is strengthened by introducing a magnetic gradient between the magnetic filter and nearby column. Because immunomagnetic beads specifically react with target cells, target cells are collected efficiently. Accumulated beads are released from the filter by removing the external magnetic force. After concentrating the samples using the HGIMS system, DNA was extracted from the samples, and PCR was applied to detect F. psychrophilum. Our primers did not react with reference bacteria and reacted specifically with F. psychrophilum. The detection sensitivity using the HGIMS system was higher than that of the method without the HGIMS system, and the total assay time, including sample preparation, was <3.5 h. PCR products of the expected size were obtained from samples of concentrated 4 × 10^?1 to 4 × 10³ cfu mL^?1F. psychrophilum more than 80% of the time using the HGIMS system. Furthermore, our proposed method could be useful for the specific detection of F. psychrophilum from actual samples. Our proposed method is suitable for the highly sensitive detection of F. psychrophilum.     

Root development and the expression of aquaporin genes in rice seedlings under osmotic stress

We conducted hydroponic culture experiments to characterize root traits in a rice cultivar ‘Puluik Arang’ that has been identified in a previous study as a cultivar that is adaptable to unflooded conditions. Root morphological traits and the expression of 11 aquaporin genes in rice seedlings (cv. Puluik Arang and cv. Akitakomachi) subjected to osmotic stress by polyethylene glycol (PEG) treatments (10 and 20%) were analysed. ‘Puluik Arang’ exhibited significantly greater water uptake under 10% PEG treatment than ‘Akitakomachi’. Lateral root development was maintained in ‘Puluik Arang’ under PEG treatments. The expression of some aquaporin genes, particularly OsTIP2;1, was higher in ‘Puluik Arang’ than in ‘Akitakomachi’. Immunocytochemical analysis showed that the OsTIP2;1 protein mainly accumulated in endodermal cells. The results suggest that better lateral root development and the function of aquaporins could contribute to water uptake in ‘Puluik Arang’ under osmotic stress.     

Immunolocalization of Pepper mild mottle virus in Capsicum annuum seeds

To clarify the mechanism of seed transmission of Pepper mild mottle virus (PMMoV), the virus was immunolocalized in Capsicum annuum seeds using fluorescence microscopy. Two distinct patterns were observed: In the first, PMMoV was present in the epidermis and parenchyma but not in the endosperm or embryo; in the second, the virus was restricted to the surface of the epidermis and parenchyma. These findings shed light on the fundamental mechanisms of seed transmission of tobamoviruses and may aid in the design of new methods to prevent the spread of seedborne virus diseases.     

不同成熟采收期对清水白桃果实达到完熟时品质的影响(英文)

以软溶质水蜜桃清水白桃为试材,分别在树上完熟的前6、4和2d采样,贮藏于25℃,相对湿度90%的环境条件下至完熟,与树上完熟果实相比较,调查了果实的生理变化、色泽和果汁品质性状,并对完熟果实进行了风味评定。结果表明,贮藏期间的果实乙烯释放量以完熟前2d采收的果实增加最快并在果实完熟时达到最大,树上完熟的果实乙烯释放量最少。树上完熟前6d采收的果实,果皮亮度、果汁的蔗糖含量、“果香型”内酯类物质含量均低于后3次采收的果实,其中以树上完熟前2d采收的果实为最高。相反滴定酸、柠檬酸、天冬酰胺和“清香型”香气物质含量以树上完熟前6d采收的果实最高。完熟果实的风味评定也显示了树上完熟前2d采收的果实综合评价最高,早期采收的果实即使贮藏后达到完熟果肉硬度,甜度和香味少、酸味重。另外,树上完熟的果实肉质粗,酸度低,有怪香味,综合评价不高。     

Effect of low molecular weight lignin fragments including oxalic acid in alkaline-oxygen stage waste liquor on Al toxicity

Our previous study indicated the existence of some low molecular weight contaminants in separated fractions (F1 to F4) of alkaline-oxygen stage waste liquor by spectroscopic analysis. In the present study, the quantities of these compounds were determined by capillary electropheresis (CE). Substantial amounts of oxalate and acetate were found in F3. The complexes between Al and oxalate as well as F3 were characterized by ²⁷Al-nuclear magnetic resonance (NMR) spectroscopy, and the spectra of Al-oxalate complexes demonstrated the ability of oxalate to chelate Al and to produce different forms of complexes at varied molar ratios. Plant growth experiments in the presence of Al-oxalate complexes suggested that at a proper range, oxalate has a favorable effect on the detoxification of Al toxicity. It can be assumed that oxalate in F3 also plays an important role in the efficiency of removing Al toxicity. ²⁷Al-NMR was proved to be a useful method for the study of complexes between Al and organic compounds without disturbing their equilibrium conditions.     

Termite resistance and wood-penetrability of chemically modified tannin and tannin-copper complexes as wood preservatives

We examined the ability of chemically modified tannin and tannin-copper complexes to penetrate wood and the ability of the treated wood to resist termites. Only the tannin-treated wood retained the agents after treatment. Wood with untreated mimosa tannin (MT) retained the least amount, followed by wood with resorcinolated tannin (RMT) and that with catecholated tannin (CMT). When RMT or CMT was mixed with ammonia-copper, the wood retained twice as much of these solutions as the MT -ammonia-copper solution. The degree of retention of RMT-NH₃-Cu and CMT-NH₃-Cu ranged from 268 to 326kg/m³. The solutions penetrated 2–13 mm from the tangential sections of the logs. We also measured the termite resistance conferred by these solutions. Most of the tannin-NH₃-Cu solutions showed contact lethality for termites in the contact toxicity test. However, the termites were fed cellulose treated with those solutions and most survived the oral toxicity test (14 days). Moreover, these solutions reduced the amount of damage to the wood by termites. However, the mortality rate of the termites during the eating-damage test (>21 days) did not reach 100% for any of the solutions except for RMT. As a result of the field stake test with the same log's used for the penetrability test, the mass loss of wood treated with RMT or CMT alone or with RMT + NH₃ + CuCl₂, was about one-third to one-half that of the controls. Because these solutions have good wood penetrability and good termite resistance, they have potential use as low-toxicity wood preservatives.     

Effect of acetic acid added to cooking water on the dissolution of proteins and activation of protease in rice

The effect of acetic acid on the dissolution of proteins in rice was studied to elucidate the mechanism for the textural change induced by the acid by chemical and SDS-PAGE analyses of the rice proteins in the soaking solution. More proteins were extracted with 0.2 M acetic acid (pH 2.7) than with water (pH 6.8). The effect of acetic acid on the protein dissolution increased with increasing temperature. Immunoblotting confirmed that, when rice was soaked in acetic acid, glutelin was dissolved into the soaking solution and degraded by aspartic proteinase. Aspartic proteinase degraded glutelin much more than it did albumin and globulin. It was found that the combined amount of albumin and globulin dissolved into the acetic acid solution was much larger than that of glutelin, despite the smaller amounts present of albumin and globulin than of glutelin. Metal ions were extracted more with acetic acid than with water. In addition, carboxypeptidase was activated under the acidic condition and resulted in an increase in the amount of free amino acids. The main effect of acetic acid on the dissolution of rice proteins was enhancement of the solubility of albumin, globulin, and glutelin, the effect of proteases being minor.     

High prevalence of Mycoplasma equirhinis in Thoroughbred horses with respiratory symptoms in autumn 2018

Mycoplasma species are often isolated from horses with respiratory symptoms; however, the pathogenicity of Mycoplasma is still unclear. In autumn of 2018, we encountered an increase in cases with respiratory symptoms, mainly coughing, in a group of Thoroughbred racehorses in Japan. We examined tracheal wash samples obtained from 40 of those cases. Bacteria and viruses that commonly cause respiratory symptoms were investigated, and anaerobes were detected in only 5 cases and Streptococcus equi subsp. zooepidemicus (S. zooepidemicus) was detected in only 1 case of 40 cases with loop-mediated isothermal amplification assay. S. zooepidemicus and Streptococcus pneumoniae were isolated at a bacterial count of higher than 1.0 × 10⁴ CFU/ml from 5 and 2 cases of 28 cases cultured, respectively. None of the viruses investigated was detected in 40 cases. Mycoplasma equirhinis (M. equirhinis) was isolated from 40.0% (16/40) of the cases, which was higher than previously reported isolation rates. The rate of M. equirhinis isolation in the cases from 2018 was significantly higher than the isolation rates in the other horses: clinical cases with respiratory symptoms in 2019–2020 (13.6%, 3/22) and healthy horses (13.5%, 5/37) in Japan. In this study, the isolation rate of M. equirhinis from horse group with cough symptoms in 2018 was high and no other common etiological agents were detected. The pathogenesis of M. equirhinis is still unclear, however, M. equirhinis might have been associated with respiratory symptoms in the Thoroughbred horse cases in 2018.     

Evaluation of antimicrobial use in dairy cattle,beef cattle and broilers in Japan using dosage-based indicators

The sales amount of antimicrobials intended for use in dairy cattle, beef cattle and broilers from 2008 to 2019 was evaluated for each antimicrobial class and administration route using dosage-based indicators. Our results revealed that the antimicrobial sales amount sold for use in dairy cattle in 2019 in terms of total weight of active ingredient, the number of defined daily doses (DDDs) (theoretical amount of biomass subjected to antimicrobial treatment in a year) and the number of treatment days (TDs) (theoretical number of days of treatment that an animal is subjected to in a year) calculated using Japanese DDD values (DDDjp values) was 36,751 kg, 8,261,848,000 kg·days and 15.5 days, respectively. Likewise, the antimicrobial sales amount sold for use in beef cattle and broilers in 2019 in terms of these metrics was 33,403 kg, 3,928,248,000 kg·days and 3.61 days, and 69,773 kg, 2,947,848,000 kg·days and 10.66 days, respectively. There was a considerable difference between the number of DDDs calculated using DDDjp values and that calculated using European DDD values (DDDvet values) for antimicrobial products sold for use in dairy and beef cattle. Our results also revealed that the sales amount of some antimicrobials, such as cephalosporins and quinolones represented larger proportions when calculated using dosage-based indicators than when calculated using the weight of active ingredient. Considering that Japanese veterinarians and farmers are more likely to conform to the Japanese dosage recommendations rather than the European ones, these results indicate the need for using dosage-based metrics, in particular metrics based on Japanese dosages rather than European dosages.