<Emphasis Type="Italic">Paralepidapedon variabile</Emphasis> sp. n. (Trematoda,Lepocreadioidea, Lepidapedidae) and other representatives of the genus <Emphasis Type="Italic">Paralepidapedon</Emphasis> from Antarctic fish

In the Ross Sea and Amundsen Sea, four representatives of the genus Paralepidapedon—Paralepidapedon cf. dubium Prudhoe et Bray 1973 sensu Sokolov et Gordeev 2013, P. lepidum (Gaevskaya et Rodyuk 1988), Paralepidapedon sp., and P. variabile sp. n.—were found in demersal fishes Muraenolepis marmorata and Macrourus whitsoni. Paralepidapedon variabile sp. n. is described from Muraenolepis marmorata in the Amundsen Sea. Paralepidapedon variabile sp. n. differs from other species of the genus Paralepidapedon by the position of the anterior border of the vitellarium at the level of the anterior edge of the ventral sucker or genital pore and by the highly variable shape of the testes: from roundish with a smooth edge to sinuate–lobate. Paralepidapedon lepidum was found for the first time in the Antarctic.     

Under-ice distribution of polar cod <Emphasis Type="Italic">Boreogadus saida</Emphasis> in the central Arctic Ocean and their association with sea-ice habitat properties

In the Arctic Ocean, sea-ice habitats are undergoing rapid environmental change. Polar cod (Boreogadus saida) is the most abundant fish known to reside under the pack-ice. The under-ice distribution, association with sea-ice habitat properties and origins of polar cod in the central Arctic Ocean, however, are largely unknown. During the RV Polarstern expedition ARK XXVII/3 in the Eurasian Basin in 2012, we used for the first time in Arctic waters a Surface and Under Ice Trawl with an integrated bio-environmental sensor array. Polar cod was ubiquitous throughout the Eurasian Basin with a median abundance of 5000 ind. km^?2. The under-ice population consisted of young specimens with a total length between 52 and 140 mm, dominated by 1-year-old fish. Higher fish abundance was associated with thicker ice, higher ice coverage and lower surface salinity, or with higher densities of the ice-amphipod Apherusa glacialis. The fish were in good condition and well fed according to various indices. Back-tracking of the sea-ice indicated that sea-ice sampled in the Amundsen Basin originated from the Laptev Sea coast, while sea-ice sampled in the Nansen Basin originated from the Kara Sea. Assuming that fish were following the ice drift, this suggests that under-ice polar cod distribution in the Eurasian Basin is dependent on the coastal populations where the sea-ice originates. The omnipresence of polar cod in the Eurasian Basin, in a good body condition, suggests that the central Arctic under-ice habitats may constitute a favourable environment for this species survival, a potential vector of genetic exchange and a recruitment source for coastal populations around the Arctic Ocean.     

Distribution pattern of Polar cod (<Emphasis Type="Italic">Boreogadus saida</Emphasis>) larvae and larval fish assemblages in relation to oceanographic parameters in the northern Bering Sea and Chukchi Sea

Polar cod, Boreogadus saida, is a key species in the Arctic Ocean ecosystem. We examined the distribution pattern of B. saida and other fish larvae in relation to oceanographic parameters, including sea surface temperature and salinity (SST and SSS), the mode of temperature and salinity within the water column (F _temp and F _sal), and the temporal duration between the date of sea ice retreat and the date of field surveys (dSRT) in the northern Bering Sea and Chukchi Sea during the summers of 2008 and 2013. Sampling was conducted onboard the T/S Oshoro-Maru using a bongo net for 2 years. At sampling stations, the temperature and salinity were measured using conductivity-temperature-depth profiler casts. We calculated dSRT from satellite derived polar gridded sea ice concentration data. A total of 1186 individuals comprising 7 families and 16 species were collected, with B. saida (35 %) and Ammodytes hexapterus (27 %) dominating the catch in number. Based on the species composition (cluster analysis), the sampling stations were divided into four groups. Pleuronectidae dominated group A, which was characterized by relatively high temperature (SST and F _temp), while B. saida dominated group B, characterized by low temperatures. A. hexapterus dominated group C and D, which had similar temperatures and salinities, but group C had a shorter dSRT than group D. The latter was also differentiated by the presence of Lumpenus sp. B. saida, which were most abundant in regions where temperatures ranged from ?2 to 0.5 °C. These results, the geographical variation of group B sampling stations, suggest there are two separate spawning areas characterized by similar fish communities and related to consistently cold bottom temperatures. Moreover, stations with a higher abundance of smaller sized larval B. saida were characterized by a short dSRT, whereas stations with a lower abundance and a larger size were characterized by a long dSRT.     

Contrasting the early life histories of sympatric Arctic gadids <Emphasis Type="Italic">Boreogadus saida</Emphasis> and <Emphasis Type="Italic">Arctogadus glacialis</Emphasis> in the Canadian Beaufort Sea

The early life stages of Boreogadus saida and Arctogadus glacialis are morphologically similar, making it difficult to assess differences in their ecological niche. The present study documented for the first time the early life stage ecology of A. glacialis, compared it to that of B. saida, and identified the factors separating the niches of the two sympatric species. The 10,565 larval gadids collected in the Beaufort Sea from April to August of 2004 and 2008 were identified to species either directly by genetics and/or otolith nucleus size, or indirectly with a redistribution procedure. Between 8.0 and 8.7 % of all gadids were assigned to A. glacialis. Larvae of A. glacialis were longer at hatch and experienced lower mortality rates than those of B. saida. The two species shared similar spatiotemporal and vertical distributions, hatching season, and growth rate. Under the ice, feeding incidence of B. saida was low (14 %) relative to A. glacialis (88 %). At lengths <15 mm, both species specialized on different prey. The diet of fish >15 mm overlapped (Schoener’s index = 0.7), with Calanus glacialis and C. hyperboreus providing >50 % of the carbon intake of both species. The higher mortality in B. saida may be explained by the smaller size at age from hatching to metamorphosis and a lower under-ice feeding incidence. The early larval stage appears to be the key period of niche divergence between the two species.     

Proteinase inhibitors from <Emphasis Type="Italic">Cajanus platycarpus</Emphasis> accessions active against pod borer <Emphasis Type="Italic">Helicoverpa armigera</Emphasis>

Cajanus platycarpus, a wild relative of Cajanus cajan, is an important source for various agronomically desirable traits, including resistance towards pod borer, Helicoverpa armigera. In the present study, the inhibitory activity of proteinase inhibitors (PIs) present in crude protein extracted from different accessions of C. platycarpus and cultivars of C. cajan was evaluated against H. armigera under in vitro and in vivo conditions. The PIs active against H. armigera gut trypsin-like proteinases (HGPs), referred to as ‘HGPIs’, were more pronounced in mature dry seeds of C. platycarpus accessions when compared with cultivars, which is also evident through gelatin activity staining studies. Therefore, the inhibitory activity of HGPIs was further evaluated in various plant organs of C. platycarpus accessions, such as leaves, flowers, pods, developing seeds at 8–10 days (DAP-I), 18–20 days (DAP-II), and 28–32 days after pollination (DAP-III). However, the HGPI activity was more pronounced in mature dry seeds > DAP-III > DAP-II > DAP-I > flowers > pods > leaves. The observed quantitative allocation of HGPIs closely resembled “Optimal Defense Theory”. Further, bioassays demonstrated that there was a significant reduction in the body weight of the larvae fed upon crude PI extracts of C. platycarpus accessions with concomitant increase in mortality rate and the formation of larval–pupal intermediates. Nevertheless, such changes were not observed when the larvae were fed on crude PI extracts of C. cajan cultivars. These results suggest that the PI gene(s) from C. platycarpus accessions could be exploited in the management of H. armigera by introgression into C. cajan cultivars.     

Compatibility and efficacy of the lady beetle <Emphasis Type="Italic">Thalassa montezumae</Emphasis> and the entomopathogenic fungus <Emphasis Type="Italic">Isaria fumosorosea</Emphasis> for biological control of the green croton scale: laboratory and greenhouse investigations

The lady beetle Thalassa montezumae and the entomopathogenic fungus Isaria fumosorosea (Ifr) were assessed alone and in combination to suppress green croton scale, Phalacrococcus howertoni, populations on croton plants using laboratory bioassays and greenhouse cage studies. The acquisition of Ifr blastospores by beetle larvae (3rd instar) and adults during contamination in well plates was used to simulate exposure to direct spraying and subsequent possible fungal infection was assessed. Spore dispersal by the insects was determined after the blastospore-contaminated T. montezumae life stages roamed on agar plates for 24 h by counting the number of colony-forming units (CFUs) produced in the plates. There were no significant differences in survival times at 14 days post-treatment between beetle larvae and adults exposed to Ifr and those exposed to water only. Mean survival time of larvae exposed to Ifr was 14 days and water 12 days, whereas for adults it was 13 days compared to 13 days, respectively. Plates with Ifr blastospore-contaminated T. montezumae adults roaming on the agar surface displayed significantly more fungal trails as CFUs compared to plates with larvae. In greenhouse cage studies, the mean mortality rates of the scale exposed to beetle larvae, either alone (80.8%) or in combination with Ifr (89.1%), were not significantly different. Scale mortality rates in the fungus-only (60.5%) and beetle larvae-only treatments were statistically similar. The treatment with both biocontrol agents had a significantly higher scale mortality rate compared to the treatment with Ifr only. Therefore, spraying Ifr prior to releasing T. montezumae is an effective and compatible biological control strategy for management of the green croton scale on croton plants.     

Molecular evidence for hybridization between invasive <Emphasis Type="Italic">Solidago canadensis</Emphasis> and native <Emphasis Type="Italic">S</Emphasis>. <Emphasis Type="Italic">virgaurea</Emphasis>

Hybridization between alien and native species is biologically very important and could lead to genetic erosion of native taxa. Solidago × niederederi was discovered over a century ago in Austria and described by Khek as a natural hybrid between the alien (nowadays regarded also as invasive) S. canadensis and native S. virgaurea. Although interspecific hybridization in the genus Solidago is considered to be relatively common, hybrid nature of S. × niederederi has not been independently proven using molecular tools, to date. Because proper identification of the parentage for the hybrid Solidago individuals solely based on morphological features can be misleading, in this paper we report an additive polymorphism pattern expressed in the ITS sequences obtained from individuals representing S. × niederederi, and confirm the previous hypothesis that the parental species of this hybrid are S. canadensis and S. virgaurea. Additionally, based on variability at the cpDNA rpl32-trnL locus, we showed that in natural populations hybridization occurs in both directions.     

Biology of a Neotropical Harlequin Stink Bug, <Emphasis Type="Italic">Runibia perspicua</Emphasis> (F.)

Laboratory and field studies were conducted with nymphs and adults of the Neotropical stink bug Runibia perspicua (F.) to evaluate their biology on plants. Total nymph mortality was ca. 13% on immature fruits of Brunfelsia australis Benth. (IFBA) and 90% on raw shelled peanuts, Arachis hypogaea L. (RSP); no nymphs survived on immature pods of green bean, Phaseolus vulgaris L., or on immature fruits of privet, Ligustrum lucidum Ait. Nymph developmental time was ca. 36 on IFBA and 55 days on RSP. Body weight was significantly greater when nymphs were raised isolated compared to those raised in groups. Adult feeding activity was greater than third instars on IFBA. Adult longevity on IFBA + cherry tomato, Solanum lycopersicum L. fruit, reached ca. 150 days. On IFBA, fecundity was 2.5 egg masses/female and 48.3 eggs/female; 25.6% of the eggs hatched. Adults did not reproduce on tomato. Body weight gain did not increase significantly for both sexes during the first four weeks of adult life on IFBA. In the field, plants of B. australis were surveyed in Passo Fundo, RS (28°15′S; 52°24′W). The majority of egg masses were deposited on the lower (abaxial) surface of leaves. Nymphs aggregated on immature fruits, and adults were seldom found on the plants. These are the first data on the performance of R. perspicua on B. australis indicating that this plant species is suitable to the bug’s biology.     

Cryopreservation of the critically endangered golden paintbrush (<Emphasis Type="Italic">Castilleja levisecta</Emphasis> Greenm.): from nature to cryobank to nature

In this study conservation of Castilleja levisecta Greenm., a globally endangered species was addressed through in vitro cryopreservation of shoot tips. In vitro cultures were successfully established using seedlings received from British Columbia, Canada. Shoot tips excised from in vitro propagated plants were cryopreserved using a droplet-vitrification method following optimization of individual protocol steps such as pre-culture, treatment with vitrification solutions, and unloading. The highest plant regrowth after cryopreservation (66%) was achieved when shoot tips were pre-cultured in 0.3 M sucrose for 17 h followed by 0.5 M sucrose for 4 h, incubated in an osmo-protectant solution (17.5% [v/v] glycerol and 17.5% [w/v] sucrose) for 20 min, exposed to vitrification solution A3 (37.5% [v/v] glycerol plus 15% [v/v] dimethylsulfoxide (DMSO) plus 15% [v/v] ethylene glycol (EG) plus 22.5% [w/v] sucrose) on ice for 40 min, and unloaded in 0.8 M sucrose solution for 30 min. Healthy plants were developed from cryopreserved shoot tips and propagated in vitro using nodal segments. Plants derived from in vitro culture and from cryopreserved tissues were successfully rooted and acclimated in a greenhouse with 100% survival rate. Acclimatized plants were reintroduced in a naturalized propagation area at the Conservation Nursery at Fort Rodd Hill, Canada. Twenty of 94 reintroduced plants (21%) survived the transit from lab to field and some had started to flower. This is the first report for cryopreservation of C. levisecta, an important step in conserving and re-introducing this critically imperiled species in nature.     

Behavioral responses of <Emphasis Type="Italic">Frankliniella occidentalis</Emphasis> to floral volatiles combined with different background visual cues

The Western flower thrips, Frankliniella occidentalis (Thysanoptera: Thripidae), is one of the most destructive sucking pests of flowering plants. We examined behavioral responses of F. occidentalis adults to the floral volatiles of Rosa chinensis, Gardenia jasminoides, and Tagetes erecta in a Y-tube olfactometer with background visual cues (green, red, white, yellow, and blue backgrounds). The results show that F. occidentalis adults had similar responses under all the different background colors, but the responses by sex varied significantly when offered floral volatile or clean air. In pairings of floral volatiles, female F. occidentalis adults presented significant preferences (R. chinensis > G. jasminoides > T. erecta) in each treatment, while male F. occidentalis adults had no significant preference. Furthermore, the responses of female F. occidentalis adults to the three different floral volatiles compared with each other were greater under green and blue backgrounds than red, white, or yellow backgrounds. However, the responses of male F. occidentalis adults to floral volatiles were similar to all five backgrounds. These results indicate that female thrips could discriminate different floral volatiles, while male thrips could not. Both male and female thrips performed steady preferences of floral volatiles under all the five visual backgrounds, and the behavioral responses of female thrips to the floral volatiles could be enhanced under the green and blue backgrounds. These findings could hopefully aid in the development of effective trapping and monitoring strategies for this pest.     

Susceptibility of <Emphasis Type="Italic">Diaphorina citri</Emphasis> (Hemiptera: Liviidae) and Its Parasitoid <Emphasis Type="Italic">Tamarixia radiata</Emphasis> (Hymenoptera: Eulophidae) to Entomopathogenic Fungi under Laboratory Conditions

Diaphorina citri (Kuwayama) is a global pest of citrus that transmits the bacteria associated with the disease, Huanglongbing. Entomopathogenic fungi and the parasitoid Tamarixia radiata (Waterston) are important biological control agents of this pest and likely to interact in D. citri populations. As a basis for interaction studies, we determined the susceptibility of nymphs and adults of D. citri and adults of the parasitoid T. radiata to six fungal isolates from the species Beauveria bassiana s.l. (Bals.-Criv.) Vuill. (isolates B1 and B3), Metarhizium anisopliae s.s. (Metsch.) (Ma129 and Ma65) and Isaria fumosorosea Wize (I2 and Pae). We conducted experiments evaluating infection levels in all three insect groups following inoculation with a series of conidial concentrations (1 × 10⁴–1 × 10⁸ conidia mL^?1). Results showed that D. citri nymphs and T. radiata were more susceptible to fungal isolates than D. citri adults. Overall, B. bassiana and M. anisopliae isolates caused the greatest infection compared with I. fumosorosea isolates in all three groups of insects. Isolates B1 (B. bassiana) and Ma129 (M. anisopliae) infected a greater proportion of adults and nymphs of D. citri, respectively. Both isolates of B. bassiana caused greater infection in T. radiata compared with isolates of the other fungal species. We propose that isolates B1 and Ma129 are the strongest candidates for control of D. citri. Our results represent the first report of entomopathogenic fungi infecting T. radiata, and the basis for future studies to design a biological control programme that uses both agents more efficiently against D. citri populations.     

Decreased microbial diversity and <Emphasis Type="Italic">Lactobacillus</Emphasis> group in the intestine of geriatric giant pandas (<Emphasis Type="Italic">Ailuropoda melanoleuca</Emphasis>)

It has been established beyond doubt that giant panda genome lacks lignin-degrading related enzyme, gastrointestinal microbes may play a vital role in digestion of highly fibrous bamboo diet. However, there is not much information available about the intestinal bacteria composition in captive giant pandas with different ages. In this study, we compared the intestinal bacterial community of 12 captive giant pandas from three different age groups (subadults, adults, and geriatrics) through PCR-denaturing gradient gel electrophoresis (DGGE) and real-time PCR analysis. Results indicated that microbial diversity in the intestine of adults was significantly higher than that of the geriatrics (p < 0.05), but not significant compared to the subadults (p > 0.05). The predominant bands in DGGE patterns shared by the twelve pandas were related to Firmicutes and Proteobacteria. Additionally, in comparison to healthy individuals, antibiotic-treated animals showed partial microbial dysbiosis. Real-time PCR analyses confirmed a significantly higher abundance of the Lactobacillus in the fecal microbiota of adults (p < 0.05), while other bacterial groups and species detected did not significantly differ among the three age groups (p > 0.05). This study revealed that captive giant pandas with different ages showed different intestinal bacteria composition.     

Chemical and molecular identification of the invasive termite <Emphasis Type="Italic">Zootermopsis nevadensis</Emphasis> (Isoptera: Archotermopsidae) in Japan

Numerous termite species have been introduced outside their native ranges by human transport, and some have become invasive. The dampwood termite Zootermopsis nevadensis (Hagen), which is native to western North America, has been introduced to and become established in Kawanishi City, Hyogo Prefecture, Japan. Zootermopsis nevadensis is subdivided into two subspecies based on cuticular hydrocarbon (CHC) phenotypes: Z. nevadensis nevadensis and Z. nevadensis nuttingi (Haverty and Thorne). Here, we identified Z. nevadensis in Japan as hybrids between the two subspecies. Chemical analysis showed the presence of 7,15-dimethylhenicosane and 5,17-dimethylhenicosane in the CHCs of Z. nevadensis in Japan, corresponding to the CHC phenotype of Z. n. nevadensis. Conversely, all mitochondrial cytochrome c oxidase subunit I sequences of Z. nevadensis in Japan were identical to sequences from Z. n. nuttingi and hybrids between the two subspecies from a native hybrid zone in California, USA. In addition, phylogenetic analysis showed that Z. nevadensis in Japan formed a clade with Z. n. nuttingi and hybrids between the two subspecies. Our results show discordance between the chemical and genetic features of Z. nevadensis in Japan, indicating that individuals of Z. nevadensis in Japan are hybrids between the two subspecies.     

Fingerprinting by amplified fragment length polymorphism (AFLP) and barcoding by three plastidic markers in the genus <Emphasis Type="Italic">Wolffiella</Emphasis> Hegelm

Amplified fragment length polymorphism (AFLP) fingerprinting and three different plastidic DNA regions (rpl16, rps16, atpF-atpH) were used to investigate species identity in the genus Wolffiella. For this purpose, clones (67 in total) belonging to all ten species were selected. Almost all the species were represented by more than one clone. The fingerprinting technique, AFLP, clearly distinguished the species, W. caudata, W. gladiata, W. neotropica, W. rotunda, and W. welwitschii. Apart from confirming the molecular identity of these five species, the plastidic markers could delineate two additional species, W. hyalina and W. denticulata, although the conclusion concerning the latter is restricted by the availability of only one clone. The efficiency of the plastid-derived markers in identifying the number of species-specific clusters followed the sequence rps16 > rpl16 > atpF-atpH. The species W. lingulata, W. oblonga, and W. repanda could not be identified by any of the molecular methods presented here, but could be strictly defined on a morphological basis. In several clones, high amounts of genetic admixtures between different species were detected. Further, simulation studies demonstrated that these clones are genetic hybrids. This might be one of the obstacles in molecular identification of species in the genus Wolffiella.     

Effect of Milk Fermented with Lactic Acid Bacteria on Diarrheal Incidence,Growth Performance and Microbiological and Blood Profiles of Newborn Dairy Calves

The effect of the administration of milk fermented with lactic acid bacteria to calves was evaluated. The strains included were: Lactobacillus murinus CRL1695, Lact. mucosae CRL1696, Lact. johnsonii CRL1693, and Lact. salivarius CRL1702, which were selected for their beneficial and functional properties and isolated from healthy calves in the northwestern region of Argentina. The trial was conducted on a dairy farm located in Tucumán (Holando-Argentino calves). A randomized controlled trial was performed in which 56 new-born animals were divided into two groups: the treated group (T) received the fermented milk for 60 days and the control group (C) only milk. The animals were fed a solid diet ad libitum. The treated group was given a daily dose of 1 × 10⁹CFU of the probiotic fermented milk while the control group was fed milk. Body weight and biometrical parameters were recorded between 15 and 60 days of age, and average daily gain was calculated with three samplings per animal throughout the trial. Rectal swabs and fecal and blood samples were also collected. Results showed the efficacy of the probiotic: lower morbidity and mortality of calves (morbidity was 69.20% in animals without the probiotic, and 46.15% in probiotic-treated animals, with P = 0.09; mortality in C was 34.61 and 7.69% in animals fed with ferment milk; P = 0.02).The calves fed with probiotic evidenced an improvement in nutritional parameters, body condition and weight gain (health index P = 0.01; average daily gain P = 0.03).Viable bacterial numbers showed no differences between the two experimental groups. Hematological parameters and serum proteins were not modified by the treatment. The results suggest that the fermented milk containing lactic acid bacteria can be a viable veterinary product for young calves due to its beneficial effects on health and growth.     

The species composition,morphology, and seasonal distribution of diatoms of the genus <Emphasis Type="Italic">Attheya</Emphasis> West, 1860 from the Sea of Japan

Six species that belong to the diatom genus Attheya were found in the Russian waters of the Sea of Japan. A. cf. flexuosa is a new record for the seas of Russia. A. decora is recorded for the first time for the Sea of Japan. The peculiarities of the morphology of A. decora and A. cf. flexuosa are described and illustrated. The study of the seasonal distribution, abundance, and ecology of Attheya species in the northwestern Sea of Japan showed that A. longicornis and A. ussurensis were the most numerous and widespread species, reaching maximum densities (up to 1.5 × 10⁶ and 1.8 × 10⁴ cells/liter, respectively) in the spring, summer, and fall. A. cf. flexuosa and A. septentrionalis were rare and were found in low numbers in the winter and early spring. Based on the ultrastructure of the girdle bands, the form and number of chloroplasts, the presence/absence of rimoportula on the valve, and habitat features, we distinguished two groups of species of the genus Attheya from the Sea of Japan. The first group includes psammophytes A. decora, A. arenicola, and A. ussurensis; the second one comprises A. longicornis and A. cf. flexuosa epiphytic on other diatoms, as well as A. septentrionalis, which is found in the under-ice plankton and is able to attach to the underside of ice.     

Insecticidal activities of monoterpenes and phenylpropenes against <Emphasis Type="Italic">Sitophilus oryzae</Emphasis> and their inhibitory effects on acetylcholinesterase and adenosine triphosphatases

In the present study, six monoterpenes [(?)-citronellal, p-cymene, (?)-menthone, α-pinene, α-terpinene, and (?)-terpinen-4-ol] and two phenylpropenes [trans-cinnamaldehyde and eugenol] were evaluated for their contact and fumigant toxicities against Sitophilus oryzae adults. The effects of these compounds on the mortality of S. oryzae adults in stored wheat and their inhibitory effects on acetylcholinesterase (AChE) and adenosine triphosphatases (ATPases) were examined. The tested compounds showed varying degrees of contact toxicity, with trans-cinnamaldehyde (LC₅₀ = 0.01 mg/cm²) being the most potent compound, followed by (?)-menthone (LC₅₀ = 0.013 mg/cm²) and eugenol (LC₅₀ = 0.015 mg/cm²). In a fumigant toxicity assay, the monoterpenes α-terpinene, p-cymene, and (?)-menthone showed the highest toxicities (LC₅₀ = 50.79, 52.37, and 54.08 μl/L air, respectively). Trans-cinnamaldehyde, (?)-citronellal, and eugenol were the least toxic (LC₅₀ > 100 μl/L air). In general, the oxygenated compounds exhibited high contact toxicities while the hydrocarbon compounds exhibited high fumigant toxicities. When tested for their insecticidal activities against S. oryzae in stored wheat, trans-cinnamaldehyde was found to be the most potent compound, with 73.9% mortality at an application rate of 0.5 g/kg and complete mortality (100%) at 1 and 5 g/kg after 1 week of treatment. All of the tested compounds showed AChE inhibition, although (?)-citronellal and trans-cinnamaldehyde presented the strongest enzyme inhibition, with IC₅₀ values of 18.40 and 18.93 mM, respectively. On the other hand, (?)-terpinene-4-ol exhibited the highest inhibition of ATPases, followed by α-pinene and α-terpinene.     

Chemical defences of native European coccinellid eggs against intraguild predation by the invasive Asian coccinellid, <Emphasis Type="Italic">Harmonia axyridis</Emphasis> (Pallas)

Harmonia axyridis (Pallas) is a coccinellid of Asian origin that has recently invaded substantial parts of Europe and is suspected to affect native coccinellid populations through intraguild predation and competition for food. Previous work has shown that two species from the Calvia genus appeared to be well protected against H. axyridis predation. To deepen our understanding on chemical protection of Calvia spp. and the predation risk by H. axyridis, we tested for susceptibility and palatability of Calvia spp. and H. axyridis eggs against predation by H. axyridis neonate larvae. Results show that eggs of C. quatuordecimguttata were mostly not eaten by H. axyridis, while eggs of the congeneric C. decemguttata were found to be largely unprotected against predation by the invasive coccinellid. We also observed that H. axyridis first instars successfully cannibalized on conspecific eggs. Removing the surface chemicals from C. quatuordecimguttata eggs resulted in significantly reduced protection from being preyed upon by H. axyridis, while applying these extracts onto C. decemguttata and H. axyridis eggs resulted in increased protection against H. axyridis larvae. The importance of surface chemicals in the interactions between H. axyridis and native coccinellids was confirmed by GC–MS analysis, showing a high diversity of hydrocarbons located on the surface of C. quatuordecimguttata eggs, i.e. more than twice as many when compared to C. decemguttata. Survival of H. axyridis larvae feeding on eggs of C. quatuordecimguttata, C. decemguttata or conspecific eggs, from which surface chemicals were removed by washing them with hexane, was not different from survival on unwashed eggs.     

Association between <Emphasis Type="Italic">cagA</Emphasis>, <Emphasis Type="Italic">vacAi</Emphasis>, and <Emphasis Type="Italic">dupA</Emphasis> genes of <Emphasis Type="Italic">Helicobacter pylori</Emphasis> and gastroduodenal pathologies in Chilean patients

In addition to the already known cagA gene, novel genetic markers have been associated with Helicobacter pylori (H. pylori) virulence: the dupA and vacAi genes. These genes might play an important role as specific markers to determine the clinical outcome of the disease, especially the vacAi gene, which has been expected to be a good marker of severe pathologies like gastric adenocarcinoma. In the present study, the association of cagA, dupA, and vacAi genes with gastroduodenal pathologies in Chilean patients was studied. One hundred and thirty-two patients positive for H. pylori were divided into two groups—non-severe and severe gastric pathologies—and investigated for the presence of cagA, dupA, and vacAi H. pylori virulence genes by PCR. The cagA gene was detected in 20/132 patients (15.2%), the vacAi1 gene was detected in 54/132 patients (40.9%), the vacAi2 gene was detected in 26/132 patients (19.7%), and the dupA gene was detected in 50/132 (37.9%) patients. Logistic regression model analysis showed that the vacAi1 isoform gene in the infected strains and the severity of the diseases outcome were highly associated, causing severe gastric damage that may lead to gastric cancer (p < 0.0001; OR = 8.75; 95% CI 3.54–21.64). Conversely, cagA (p = 0.3507; OR = 1.62; 95% CI 0.59–4.45) and vacAi2 (p = 0.0114; OR = 3.09; 95% CI 1.26–7.60) genes were not associated with damage, while the dupA gene was associated significantly with non-severe clinical outcome (p = 0.0032; OR = 0.25; 95% CI 0.09–0.65). In addition, dupA gene exerts protection against severe gastric pathologies induced by vacAi1 by delaying the outcome of the disease by approximately 20 years.     

Control effect on the brown-marmorated stink bug, <Emphasis Type="Italic">Halyomorpha halys</Emphasis> (Hemiptera: Pentatomidae), by combined spraying of pyrethroid and neonicotinoid insecticides in apple orchards in northern Japan

Control of Halyomorpha halys (Stål) adults by combined spraying of pyrethroid and neonicotinoid insecticides has been investigated in apple orchards in northern Japan in 2008 and 2009. Insecticides were selected to control not only H. halys but also the common lepidopteran pests Carposina sasakii Matsumura, Adoxophyes orana fasciata Walsingham, and Phyllonorycter ringoniella Matsumura. Dinotefuran was sprayed in mid-May and early July, fenpropathrin in late June, and bifenthrin in late July. H. halys adults were released into apple trees at 3–5-day intervals from mid-May to mid-August. In this spraying program, survival of the adults increased 6 days after spraying, but relatively few injury marks were noted throughout the growing season. On the other hand, in the conventional spraying program, the survival ratio of the adults increased rapidly 3 days after spraying—especially after the organophosphate applied in mid-May, late June, and late July—and increases in the number of injury marks synchronized well with survival in both years. This spray program reduced the number of fruit injury marks to one-fifth that of the conventional spraying program and one-tenth that of fungicides alone.