The development of Spodoptera frugiperda (Lepidoptera: Noctuidae) on 8 major crops in Japan and its oviposition preference for maize and soybean.

The fall armyworm, Spodoptera frugiperda (J.E. Smith), is a polyphagous pest native to the Americas. It first invaded Japan in July 2019, and the most damaged crop in this region to date has been maize. To determine pest control strategies, the range of crops potentially damaged by S. frugiperda in Japan should be identified. In this study, the effects of 8 major crops in Japan-forage maize Zea mays subsp. mays, sugarcane Saccharum officinarum, forage and edible rice Oryza sativa subsp. japonica, soybean Glycine max, eggplant Solanum melongena, green bell pepper Capsicum annuum var. grossum, sweet potato Ipomoea batatas, and taro Colocasia esculenta-on the development of S. frugiperda were analyzed by feeding leaves of domestic cultivars during larval stage. Spodoptera frugiperda developed from hatching to adulthood and laid eggs in all 7 treatments, except for taro. However, among them, only soybean showed comparable developmental suitability to maize. Therefore, its oviposition preference for maize and soybean was examined using further nonchoice and choice tests. In the nonchoice test, the number of egg masses oviposited for 3 days on soybean plants was significantly less than that on test containers, while that on maize was comparable to that on containers. These findings can explain partly why major damage has been limited to maize in Japan.

Diagnostic Performance of BD Phoenix CPO Detect Panels for Detection and Classification of Carbapenemase-Producing Gram-Negative Bacteria.

BD Phoenix CPO Detect panels can identify and classify carbapenemase-producing organisms (CPOs) simultaneously with antimicrobial susceptibility testing (AST) for Gram-negative bacteria. Detection and classification of carbapenemase producers were performed using the BD Phoenix CPO Detect panels NMIC/ID-441 for Enterobacterales, NMIC/ID-442 for nonfermenting bacteria, and NMIC-440 for both. The results were compared with those obtained using comparator methods. A total of 133 strains (32 Klebsiella pneumoniae, 37 Enterobacter cloacae complex, 33 Pseudomonas aeruginosa, and 31 Acinetobacter baumannii complex strains), including 60 carbapenemase producers (54 imipenemases [IMPs] and 6 OXA type), were analyzed. Using panels NMIC-440 and NMIC/ID-441 or NMIC/ID-442, all 54 IMP producers were accurately identified as CPOs (positive percent agreement [PPA], 100.0%; 54/54). Among the 54 IMP producers identified as CPOs using panels NMIC-440 and NMIC/ID-441, 12 and 14 Enterobacterales were not resistant to carbapenem, respectively. Among all 54 IMP producers, 48 (88.9%; 48/54) were correctly classified as Ambler class B using panel NMIC-440. Using panels NMIC-440 and NMIC/ID-442, all four OXA-23-like carbapenemase-producing A. baumannii complex strains (100.0%, 4/4) were correctly identified as CPOs, and three (75.0%, 3/4) were precisely classified as class D using panel NMIC-440. Both carbapenemase producers harboring the blaISAba1-OXA-51-like gene were incorrectly identified as non-CPOs using panels NMIC-440 and NMIC/ID-442. For detecting carbapenemase producers, the overall PPA and negative percent agreement (NPA) between panel NMIC-440 and the comparator methods were 96.7% (58/60) and 71.2% (52/73), respectively, and the PPA and NPA between panels NMIC/ID-441 or NMIC/ID-442 and the comparator methods were 96.7% (58/60) and 74.0% (54/73), respectively. BD Phoenix CPO Detect panels can successfully screen carbapenemase producers, particularly IMP producers, regardless of the presence of carbapenem resistance and can be beneficial in routine AST workflows. IMPORTANCE Simple and efficient screening methods of detecting carbapenemase producers are required. BD Phoenix CPO Detect panels effectively screened carbapenemase producers, particularly IMP producers, with a high overall PPA. As the panels enable automatic screening for carbapenemase producers simultaneously with AST, the workflow from AST to confirmatory testing for carbapenemase production can be shortened. In addition, because carbapenem resistance varies among carbapenemase producers, the BD Phoenix CPO Detect panels, which can screen carbapenemase producers regardless of carbapenem susceptibility, can contribute to the accurate detection of carbapenemase producers. Our results report that these panels can help streamline the AST workflow before confirmatory testing for carbapenemase production in routine microbiological tests.

Synergistic Effects of the Red Light and Blue Traps on Control of Thrips palmi (Thysanoptera: Thripidae).

The melon thrips (Thrips palmi) is a serious insect pest of vegetables and ornamental plants. To control and monitor this thrips, blue traps are widely used. We previously reported that irradiation of cucumber plants with red light-emitting diodes (660 nm) causes T. palmi to avoid the plants. Here, we evaluated the responses of T. palmi adults to a cucumber plant in an experimental arena with either a transparent sticky trap or a blue sticky trap, without or with illumination by a red LED panel. When T. palmi were introduced into the arena, the number of thrips attracted to the plant was lowest in treatments using the red light and blue trap. On the other hand, when T. palmi were inoculated on plants first, most thrips stayed put on the plants. Placing both red LEDs and blue sticky traps in greenhouses before the thrips invade the greenhouse from the outside might effectively control T. palmi.

Expression of a gene for an MLX56 defense protein derived from mulberry latex confers strong resistance against a broad range of insect pests on transgenic tomato lines.

Insect pests cause serious damage in crop production, and various attempts have been made to produce insect-resistant crops, including the expression of genes for proteins with anti-herbivory activity, such as Bt (Bacillus thuringiensis) toxins. However, the number of available genes with sufficient anti-herbivory activity is limited. MLX56 is an anti-herbivory protein isolated from the latex of mulberry plants, and has been shown to have strong growth-suppressing activity against the larvae of a variety of lepidopteran species. As a model of herbivore-resistant plants, we produced transgenic tomato lines expressing the gene for MLX56. The transgenic tomato lines showed strong anti-herbivory activities against the larvae of the common cutworm, Spodoptera litura. Surprisingly, the transgenic tomato lines also exhibited strong activity against the attack of western flower thrips, Frankliniera occidentalis. Further, growth of the hadda beetle, Henosepilachna vigintioctopunctata, fed on leaves of transgenic tomato was significantly retarded. The levels of damage caused by both western flower thrips and hadda beetles were negligible in the high-MLX56-expressing tomato line. These results indicate that introduction of the gene for MLX56 into crops can enhance crop resistance against a wide range of pest insects, and that MLX56 can be utilized in developing genetically modified (GM) pest-resistant crops.

α-Ionone, an Apocarotenoid, Induces Plant Resistance to Western Flower Thrips, Frankliniella occidentalis, Independently of Jasmonic Acid.

Apocarotenoids, such as ß-cyclocitral, α-ionone, ß-ionone, and loliolide, are derived from carotenes via chemical or enzymatic processes. Recent studies revealed that ß-cyclocitral and loliolide play an important role in various aspects of plant physiology, such as stress responses, plant growth, and herbivore resistance. However, information on the physiological role of α-ionone is limited. We herein investigated the effects of α-ionone on plant protection against herbivore attacks. The pretreatment of whole tomato (Solanum lycopersicum) plants with α-ionone vapor decreased the survival rate of western flower thrips (Frankliniella occidentalis) without exhibiting insecticidal activity. Exogenous α-ionone enhanced the expression of defense-related genes, such as basic ß-1,3-glucanase and basic chitinase genes, in tomato leaves, but not that of jasmonic acid (JA)- or loliolide-responsive genes. The pretreatment with α-ionone markedly decreased egg deposition by western flower thrips in the JA-insensitive Arabidopsis (Arabidopsis thaliana) mutant coi1-1. We also found that common cutworm (Spodoptera litura) larvae fed on α-ionone-treated tomato plants exhibited a reduction in weight. These results suggest that α-ionone induces plant resistance to western flower thrips through a different mode of action from that of JA and loliolide.

Loliolide, a Carotenoid Metabolite, Is a Potential Endogenous Inducer of Herbivore Resistance.

Jasmonic acid (JA) plays an important role in the induction of herbivore resistance in many plants. However, JA-independent herbivore resistance has been suggested. An herbivore-resistance-inducing substance was isolated from Tobacco mosaic virus-infected tobacco (Nicotiana tabacum) leaves in which a hypersensitive response (HR) was induced and identified as loliolide, which has been identified as a ß-carotene metabolite. When applied to tomato (Solanum lycopersicum) leaves, loliolide decreased the survival rate of the two-spotted spider mite, Tetranychus urticae, egg deposition by the same pest, and the survival rate of larvae of the common cutworm Spodoptera litura without exhibiting toxicity against these herbivores. Endogenous loliolide levels increased not only with an infestation by S litura larvae, but also with the exogenous application of their oral secretions in tomato. A microarray analysis identified cell-wall-associated defense genes as loliolide-responsive tomato genes, and exogenous JA application did not induce the expression of these genes. Suppressor of zeaxanthin-less (szl), an Arabidopsis (Arabidopsis thaliana) mutant with a point mutation in a key gene of the ß-carotene metabolic pathway, exhibited the decreased accumulation of endogenous loliolide and increased susceptibility to infestation by the western flower thrip (Frankliniella occidentalis). A pretreatment with loliolide decreased susceptibility to thrips in the JA-insensitive Arabidopsis mutant coronatine-insensitive1 Exogenous loliolide did not restore reduced electrolyte leakage in szl in response to a HR-inducing bacterial strain. These results suggest that loliolide functions as an endogenous signal that mediates defense responses to herbivores, possibly independently of JA, at least in tomato and Arabidopsis plants.

In Vitro Activity of Lascufloxacin against Streptococcus pneumoniae with Mutations in the Quinolone Resistance-Determining Regions.

Lascufloxacin showed potent activity against Streptococcus pneumoniae with a GyrA or ParC mutation (first-step mutant). The frequency of selecting resistant strains tended to be lower for lascufloxacin than for levofloxacin and garenoxacin after drug exposure in first-step mutants but was similar in the comparison between lascufloxacin and moxifloxacin. The increase in MIC was smaller for lascufloxacin than for levofloxacin, garenoxacin, and moxifloxacin when clinical strains with only ParC mutations were exposed to the corresponding drug.

Fluoroquinolone resistance in extended-spectrum ß-lactamase-producing Klebsiella pneumoniae in a Japanese tertiary hospital: silent shifting to CTX-M-15-producing K. pneumoniae.

PURPOSE: Fluoroquinolone resistance (FQ-r) in extended-spectrum ß-lactamase (ESBL) producers is an urgent health concern in countries where ESBL-producing K. pneumoniae (ESBL-Kpn) is prevalent. We investigated FQ-r in Japan where ESBL-Kpn is less prevalent. METHODOLOGY: Clinical ESBL-Kpn isolates from 2011 to 2013 were collected in Nagasaki University Hospital. The ESBL genotypes included CTX-M-15, and the mechanisms of FQ-r through plasmid-mediated quinolone resistance (PMQR) and mutations in quinolone resistance-determining regions (QRDRs) were examined. Clonality was analysed by enterobacterial repetitive intergenic consensus (ERIC)-PCR and multi-locus sequence typing was performed on selected isolates.Results/Key findings. Thirty ESBL-Kpn isolates, including seven levofloxacin-resistant isolates, were obtained from different patients. An increase in CTX-M-15-producing strains was observed during the study period (0/11 in 2011, 3/8 in 2012, and 5/11 in 2013). PMQR was detected in 53.3 % of the isolates and aac-(6')-Ib-cr was the most common (36.7 %). ST15 was observed in 60.0 % of the isolates, and for the most predominant ERIC-PCR profiles, 62.5 % of the isolates possessed the CTX-M-15 genotype and 71.4 % were levofloxacin-resistant. Levofloxacin-resistance was significantly more common in CTX-M-15 isolates (62.5 %) compared to non-CTX-M-15 isolates (9.1 %). Three QRDR mutations and aac(6')-Ib-cr, but not qnrB and qnrS, were significantly enriched in the CTX-M-15 isolates (100.0 %) compared to the non-CTX-M-15 isolates (13.6 %). CONCLUSION: Cumulatively, these results indicate that the epidemic strain, the CTX-M-15-producing K. pneumoniae ST15, is covertly spreading even when ESBL producers are not prevalent. Monitoring these epidemic strains and ESBLs in general is important for quickly identifying health crises and minimizing future risks from FQ-r ESBL-Kpn.

The Rapid Induction of Carbapenem-Resistance in an Aeromonas dhakensis Blood Isolate.

Meropenem-susceptible and -resistant Aeromonas dhakensis isolates from blood cultures of a fatal case of septicemia were analyzed. The two isolates were homologous and gene expression of metallo-ß-lactamase in the resistant strain was upregulated. Physicians should be aware of the possibility of the induction of carbapenem-resistance, following the use of carbapenems in the treatment of Aeromonas infection.

[Diagnostic Utilities of an Automated and Standardized DNA Quantification during Cytomegalovirus Monitoring].

The accurate and standardized diagnosis of cytomegalovirus (CMV) infection is important for immunocom- promised patients. We prospectively evaluated the performance of an automated and standardized real-time polymerase chain reaction (PCR) -based DNA quantification for the detection of CMV. The results of PCR- based analysis were also compared with pp65 antigenemia (Ag) assay in the clinical records. The PCR- based analysis of 144 plasma samples from 26 patients with hematologic diseases detected CMV in 69 (48.0%) samples (range, <150-1.28 X 104 copies/mL) while Ag detected CMV in 32(22.2%) samples (range, 1-37/50,000 cells). The number of concordant samples between the two tests was 95(66.0%). There were nine patients who had an Ag-positive period sandwiched by Ag-negative periods and, in all these patients, the Ag-positive period was completely covered by PCR-positive period. These results suggest that PCR can detect CMV more sensitively than Ag. The automated and standardized PCR for detection of CMV can support the appropriate management in patients with risks of CMV infection. [Original].

Hormonal regulation and developmental role of Krüppel homolog 1, a repressor of metamorphosis, in the silkworm Bombyx mori.

Juvenile hormone (JH) has an ability to repress the precocious metamorphosis of insects during their larval development. Krüppel homolog 1 (Kr-h1) is an early JH-inducible gene that mediates this action of JH; however, the fine hormonal regulation of Kr-h1 and the molecular mechanism underlying its antimetamorphic effect are little understood. In this study, we attempted to elucidate the hormonal regulation and developmental role of Kr-h1. We found that the expression of Kr-h1 in the epidermis of penultimate-instar larvae of the silkworm Bombyx mori was induced by JH secreted by the corpora allata (CA), whereas the CA were not involved in the transient induction of Kr-h1 at the prepupal stage. Tissue culture experiments suggested that the transient peak of Kr-h1 at the prepupal stage is likely to be induced cooperatively by JH derived from gland(s) other than the CA and the prepupal surge of ecdysteroid, although involvement of unknown factor(s) could not be ruled out. To elucidate the developmental role of Kr-h1, we generated transgenic silkworms overexpressing Kr-h1. The transgenic silkworms grew normally until the spinning stage, but their development was arrested at the prepupal stage. The transgenic silkworms from which the CA were removed in the penultimate instar did not undergo precocious pupation or larval-larval molt but fell into prepupal arrest. This result demonstrated that Kr-h1 is indeed involved in the repression of metamorphosis but that Kr-h1 alone is incapable of implementing normal larval molt. Moreover, the expression profiles and hormonal responses of early ecdysone-inducible genes (E74, E75, and Broad) in transgenic silkworms suggested that Kr-h1 is not involved in the JH-dependent modulation of these genes, which is associated with the control of metamorphosis.

[The Test Performance and the Differences between Sites of Blood Collection Using the Point of Care Testing-Enabled Blood Glucose Analyzer].

The point-of-care testing (POCT) has been widely performed as decentralized laboratory testing at the site of patient care. Antsense ROSE (ROSE), a POCT device for measurement of blood glucose, is handheld and time-saving and reduces risks of infection because disposable tips are used. In this study, we evaluated the testing performance of ROSE. This study was approved by the ethics committee of Nagasaki University Graduate School of Biomedical Sciences. Clinical blood samples were used for evaluating the reproducibility of testing, the calibration stability, the linearity of dilution, and the effects of dissolved oxygen partial pressure hematocrit, coexisting materials, and anticoagulant agents on the results of ROSE. Blood glucose values during 75g oral glucose tolerance testing (OGTT) were measured for analysis of the differences between sites of blood collection. Comparing with the automatic analyzer based on the hexokinase assay, the accuracy of blood glucose values measured by ROSE was evaluated. ROSE showed good performances in reproducibility, calibration stability, and dilution linearity. The values measured by ROSE were stable to oxygen partial pressure, hematocrit, coexisting materials, and anticoagulant agents and were correlated with those by the automatic analyzer. The values in capillary blood obtained from the ear or the finger elevated as approximately 30-70% as those in venous blood, at 0.5 and 1 hour after OGTT. The inhibitory effect by sodium fluoride (NaF) was 5-25% decrease after 24 hours. In conclusion, ROSE provides the blood glucose values accurately and is a rapid and useful device as a POCT.

Determination by LC-MS of juvenile hormone titers in hemolymph of the silkworm, Bombyx mori.

Juvenile hormone (JH) I, II and III in the hemolymph of the silkworm, Bombyx mori were quantified by liquid chromatography-mass spectrometry (LC-MS). JHs were treated with methanol and trifluoroacetic acid to convert into JH methoxyhydrines (JH-MHs). The key to the analytical condition for JH-MHs was the addition of 5 µM sodium acetate to the eluting solution. Each JH-MH was observed as the sodium adduct ion with good sensitivity. This improved method enabled the titration of JH I, II and III in hemolymph of the silkworm to be monitored from the 3rd instar through to the early pupal stage. A peak of JH I was observed immediately after ecdysis in the 3rd and 4th instar stages. The JH I titer sharply decreased on day 1 and reached the lowest level before ecdysis, but there was no peak at the beginning of the 5th stadium, and no apparent increase was observed until pupation.

A comparison of artificial diets for mass rearing Brontispa longissima (Coleoptera: Chrysomelidae) as hosts for the larval and the pupal parasitoids.

The coconut hispine beetle, Brontispa longissima (Gestro) (Coleoptera: Chrysomelidae) is a serious pest of coconut palm. In this study, we developed an artificial diet for B. longissima so that the beetle could be used as a host for rearing two of its parasitoids, Asecodes hispinarum Boucek (Hymenoptera: Eulophidae) and Tetrastichus brontispae Ferrière (Hymenoptera: Eulophidae). The new artificial diet represents an improvement of our previous diet, which we used as a control. When beetle larvae were reared on the new diet, which contains cysteine but not cellulose powder and has twice as much coconut leaf powder as in the control, the adult emergence was 71% (approximately 2 times that in the control). We also examined the suitability of beetles fed on the new diet as hosts for the larval parasitoid A. hispinarum and the pupal parasitoid T. brontispae. The percentage of wasps that emerged from hosts that were fed the new diet was higher than that from the control-fed hosts. The new diet allowed both A. hispinarum and T. brontispae to produce adult wasps of the next generation, whereas the control only allowed T. brontispae to produce the next generation. These results suggest that the new diet is suitable for B. longissima and will facilitate mass-rearing of A. hispinarum and T. brontispae.

Infestation and development of Sitophilus spp. in pouch-packaged spaghetti in Japan.

Infestation of pouch-packaged spaghetti by Sitophilus oryzae (L.) (Coleoptera: Curculionidae) was observed in Japan in May 2006. It was suggested that 30% of all shipped pasta products sustain damage to the packages heat-sealed before reaching retail stores. Hence, we investigated the mechanisms by which S. oryzae and Sitophilus zeamais Motschulsky (Coleoptera: Curculionidae) adults invade pasta packaged in polypropylene pouches. In the damaged packages, both species entered through the breaks in the seal and laid their eggs on the pasta. However, in the intact packages, the adults of both species bit around air vent holes in the film, but they did not make the holes big enough for entry into the pouch. We also compared each species' developmental periods from oviposition to emergence in pasta and brown rice (Oryza spp.). The average developmental period of both species at 25 degrees C was shorter in rice than in pasta. The number of individuals emerging from brown rice was significantly higher than that from pasta. Finally, we suggested a measure to prevent infestation by insect pests in pouch-packaged pasta products.

Flight capability and fatty acid level in triacylglycerol of long-distance migratory adults of the common cutworm, Spodoptera litura.

The larvae of Spodoptera litura were reared on an artificial diet, and the flight capability, and triacylglycerol (TG) level plus its fatty acid composition in 3-day-old sexually mature and non-fed adults were compared. In males, during 3 hr of tethered flight, the levels of abdominal TG and its fatty acid components did not change. But thereafter the TG and fatty acids, significantly unsaturated fatty acids in TG declined in their levels with the prolongation of flight, unsaturated fatty acids being exhausted preceding saturated fatty acid decline. When males were tested by tethered flight for 20 hr, some could fly for nearly the whole period, and were judged to be able to fly for approximately 24 hr, depending on the level of residual TG. Fatty aids in TG decreased in females similarly to males during tethered flight and some females with fully developed ovaries deposited eggs after 12 hr of flight similarly to non-flown individuals, which supports the long-distance flight capability even in sexually mature females. These results are discussed with regard to the overseas migration of this moth.