Monitoring cucumber beetles in sweetpotato and cucurbits with kairomone-baited traps

Seven kairomone formulations (Trécé, Inc., Salinas, CA) were evaluated for their effectiveness as attractants for luring three species of cucumber beetles into Pherocon CRW traps (Trécé, Inc.) in cucurbit and sweetpotato fields. The spotted cucumber beetle, Diabrotica undecimpunctata howardi Barber; the banded cucumber beetle, Diabrotica balteata LeConte; and the striped cucumber beetle, Acalymma vittatum (F.), were captured in this study. TRE8276 (TIC mixture: 500 mg of 1,2,4-trimethoxybenzene, 500 mg of indole, and 500 mg of trans-cinnamaldeyde) and TRE8336 (500 mg of 1,2,4-trimethoxybenzene, 500 mg of trans-cinnamaldeyde, 500 mg of 4-methoxyphenethanol) were the most effective lures for spotted and striped cucumber beetles. None of the kairomone lures was very effective for attracting banded cucumber beetles. Three population peaks of spotted cucumber beetles were observed in cucurbit and sweetpotato fields at the U.S. Vegetable Laboratory (Charleston, SC). The efficacy of TRE8276 declined rapidly after 2 wk in the field. An improved design of the Pherocon CRW trap, with a yellow bottom and more-tapered top section, was more effective for capturing cucumber beetles than the original trap design made entirely of clear plastic. Banded cucumber beetles were not captured in sweetpotato fields at inland locations in North Carolina or South Carolina.     

Ophiostomatoid fungi including two new fungal species associated with pine root-feeding beetles in northern Spain

Many bark beetles live in a symbiosis with ophiostomatoid fungi but very little is known regarding these fungi in Spain. In this study, we considered the fungi associated with nine bark beetle species and one weevil infesting two native tree species (Pinus sylvestris and Pinus nigra) and one non-native (Pinus radiata) in Cantabria (Northern Spain). This included examination of 239 bark beetles or their galleries. Isolations yielded a total of 110 cultures that included 11 fungal species (five species of Leptographium sensu lato including Leptographium absconditum sp. nov., five species of Ophiostoma sensu lato including Ophiostoma cantabriense sp. nov, and one species of Graphilbum). The most commonly encountered fungal associates of the bark beetles were Grosmannia olivacea, Leptographium procerum, and Ophiostoma canum. The aggressiveness of the collected fungal species was evaluated using inoculations on two-year-old P. radiata seedlings. Leptographium wingfieldii, Leptographium guttulatum, and Ophiostoma ips were the only species capable of causing significant lesions.     

Rapid identification and detection of pine pathogenic fungi associated with mountain pine beetles by padlock probes

Fifteen million hectares of pine forests in western Canada have been attacked by the mountain pine beetle (Dendroctonus ponderosae; MPB), leading to devastating economic losses. Grosmannia clavigera and Leptographium longiclavatum, are two fungi intimately associated with the beetles, and are crucial components of the epidemic. To detect and discriminate these two closely related pathogens, we utilized a method based on ligase-mediated nucleotide discrimination with padlock probe technology, and signal amplification by hyperbranched rolling circle amplification (HRCA). Two padlock probes were designed to target species-specific single nucleotide polymorphisms (SNPs) located at the inter-generic spacer 2 region and large subunit of the rRNA respectively, which allows discrimination between the two species. Thirty-four strains of G. clavigera and twenty-five strains of L. longiclavatum representing a broad geographic origin were tested with this assay. The HRCA results were largely in agreement with the conventional identification based on morphology or DNA-based methods. Both probes can also efficiently distinguish the two MPB-associated fungi from other fungi in the MPB, as well as other related fungi in the order Ophiostomatales. We also tested this diagnostic method for the direct detection of these fungi from the DNA of MPB. A nested PCR approach was used to enrich amplicons for signal detection. The results confirmed the presence of these two fungi in MPB. Thus, the padlock probe assay coupled with HRCA is a rapid, sensitive and reproducible method for the identification and detection of these ophiostomatoid fungi.     

Unexpected visitors: flightless beetles in window traps

Ancient woodlands are highly important for biodiversity. Flightless beetles and window traps may first appear to be incompatible as a study group and a study method. Little is known about the actual capability of window traps in monitoring of entire forest arthropod communities. We try to determine the possibility of using window traps (designed as tree traps) as an additional method to trap flightless beetles of the genera Carabus (Carabidae), Acalles and Trachodes (Curculionidae), as vulnerable representatives of wildlife forest fauna with high conservation value. From the environmental point of view, the occurrence of the studied flightless beetles was significantly more influenced by altitude than tree species as the studied environmental variables. Several beetles also showed positive significant species to species relationships during their observed phenological activity. Our results showed that the studied flightless beetles may reappear in secondary forests if these reach their optimal structure. On the other hand, they may be sensitive to large scale disturbances and not only to those caused by commercial forestry. The results also showed that window traps designed as tree traps could be used to successfully trap assemblages of forest fauna including flightless species.     

Identification of fungi associated with municipal compost using DNA-based techniques

Fungi are important in terrestrial decay processes. However, fungi associated with organic decay during composting are still not well known. In this study culture-independent methods were used to identify fungi associated with composting organic municipal wastes to gain a better understanding of the diversity of fungi associated with this process. Fungal communities from 0, 210, and 410 day-old compost samples were assessed with DNA fingerprinting using denaturing gradient gel electrophoresis (DGGE) and by the analysis of DNA sequences from rDNA clone libraries. From 207 rDNA sequences, 82 fungal OTU’s were detected. A disproportionate number of yeast sequences were detected in Day 0 clone libraries, including the human pathogens Candida tropicalis and Candida krusei (Saccharomycetales). Basidiomycetes accounted for over half of the clones from the Day 210 sample. Clones of Cercophora and Neurospora species accounted for most of the fungal clones of the Day 410 sample. No Zygomycetes or Aspergillus species were detected in this study. These findings call for a reassessment of long held views about the organisms involved in the composting of organic municipal wastes.     

Nitrogen-fixing bacteria associated with bark beetles

Nitrogen-fixingEnterobacter agglomerans andEnterobacter spp. were consistently isolated from the bark beetleDendroctonus terebrans. Large populations of nitrogen-fixing bacteria were found with the beetle, although no in situ acetylene reduction was demonstrated. The constant occurrence of nitrogenfixing bacteria withD. terebrans suggests a symbiotic relationship. Nitrogen-fixing bacteria were also isolated from the bark beetlesDendroctonus frontalis andIps avulsus.Mention of trade names is not an endorsement by the U.S. Department of Agriculture.     

Ophiostomatoid fungi associated with hardwood-infesting bark and ambrosia beetles in Poland: Taxonomic diversity and vector specificity

Fungi in the orders Ophiostomatales and Microascales (Ascomycota), often designated as ophiostomatoid fungi, are frequent associates of scolytine bark and ambrosia beetles that colonize hardwood and coniferous trees. Several species, e.g., Ophiostoma novo-ulmi, are economically damaging pathogens of trees. Because little is known regarding the ophiostomatoid fungi in Europe, we have explored the diversity of these fungi associated with hardwood-infesting beetles in Poland. This study aims to clarify the associations between fungi in the genera Ambrosiella, Graphium (Microascales), Graphilbum, Leptographium, Ophiostoma and Sporothrix (Ophiostomatales) and their beetle vectors in hardwood ecosystems. Samples associated with 18 bark and ambrosia beetle species were collected from 11 stands in Poland. Fungi were isolated from adult beetles and galleries. Isolates were identified based on morphology, DNA sequence comparisons for five gene regions (ITS, LSU, ßT, TEF 1-α, and CAL) and phylogenetic analyses. In total, 36 distinct taxa were identified, including 24 known and 12 currently unknown species. Several associations between fungi and bark and ambrosia beetles were recorded for the first time. In addition, associations between Dryocoetes alni, D. villosus, Hylesinus crenatus, Ernoporus tiliae, Pteleobius vittatus and ophiostomatoid fungi were reported for the first time, and Sporothrix eucastanea was reported for the first time outside of the USA. Among the species of Ophiostomatales, 14 species were in Ophiostoma s. l., two species were in Graphilbum, nine species were in Sporothrix, and seven species were in Leptographium s. l. Among the species of Microascales, three species were in Graphium, and one was in Ambrosiella. Twenty taxa were present on the beetles and in the galleries, twelve only on beetles, and four only in galleries. Bark and ambrosia beetles from hardwoods appear to be regular vectors, with ophiostomatoid fungi present in all the beetle species. Most ophiostomatoid species had a distinct level of vector/host specificity, although Ophiostoma quercus, the most frequently encountered species, also had the greatest range of beetle vectors and tree hosts. Plant pathogenic O. novo-ulmi was found mainly in association with elm-infesting bark beetles (Scolytus multistriatus, S. scolytus, and P. vittatus) and occasionally with H. crenatus on Fraxinus excelsior and with Scolytus intricatus on Quercus robur.     

危害松树的小蠹虫与其伴生茵的相互关系

危害健康松属植物的小蠹虫经常与一些特殊的真菌相联系.在小蠹虫危害松属植物的过程中,这些真菌被小蠹虫的一些特殊结构或者体表携带到松属植物上.小蠹虫与其伴生菌的联系表明小蠹虫和其伴生菌之间是一种互惠互利的关系.伴生菌随小蠹虫扩散而被带到新的寄主树木.而伴生菌或作为小蠹虫的食物来源,但更重要的是,有些伴生菌能够通过其菌丝渗透寄主组织,释放毒素,致死寄主树木,以帮助小蠹虫降低寄主抗性.许多研究致力于探索小蠹虫/伴生菌联合体与寄主树木之间关系的特征和确定小蠹虫与其伴生菌相互关系在生态学上的意义.然而,不同小蠹虫和其伴生菌所组成的共生体系,不同小蠹虫的种群数量,和不同环境条件下同种小蠹虫与其伴生菌相互作用方式的差异使我们在研究小蠹虫和其伴生菌这个共生体系时,对它们各自在成功聚集寄主树木过程中所发挥的重要作用的概括变得非常困难.     

Use of infochemicals to attract carrion beetles into pitfall traps

When the bodies of small vertebrates start to decay shortly after death, a number of organosulfur compounds are produced, including methanethiol, dimethylsulfide (DMS), dimethyldisulfide (DMDS), dimethyltrisulfide (DMTS), and S-methyl thioacetate. These molecules appear to attract various necrophagous animals. We tested the roles of DMS, DMDS, and DMTS (in order of decreasing volatility) as attractants of carrion beetles (Coleoptera: Silphidae: Nicrophorinae) in a field experiment in an agricultural landscape in southern Bohemia, Czech Republic. We collected a total of 362 adult Nicrophorus vespillo (L.) that were attracted to 220 baited pitfall traps in a 3-day experiment. Sets of traps baited with DMTS were more successful in catching N. vespillo than sets baited with a blank. Traps containing DMDS had higher trapping success than traps containing DMS. In addition, trapping success strongly increased using DMTS in the presence of DMDS but not of DMS, suggesting a synergistic effect of DMDS and DMTS. We observed similar patterns between males and females in response to the infochemicals tested.     

危害松树的小蠹虫与其伴生菌的相互关系

危害健康松属植物的小蠹虫经常与一些特殊的真菌相联系。在小蠹虫危害松属植物的过程中,这些真菌被小蠹虫的一些特殊结构或者体表携带到松属植物上。小蠹虫与其伴生菌的联系表明小蠹虫和其伴生菌之间是一种互惠互利的关系。伴生菌随小蠹虫扩散而被带到新的寄主树木。而伴生菌或作为小蠹虫的食物来源,但更重要的是,有些伴生菌能够通过其菌丝渗透寄主组织,释放毒素,致死寄主树木,以帮助小蠹虫降低寄主抗性。许多研究致力于探索小蠹虫/伴生菌联合体与寄主树木之间关系的特征和确定小蠹虫与其伴生菌相互关系在生态学上的意义。然而,不同小蠹虫和其伴生菌所组成的共生体系,不同小蠹虫的种群数量,和不同环境条件下同种小蠹虫与其伴生菌相互作用方式的差异使我们在研究小蠹虫和其伴生菌这个共生体系时,对它们各自在成功聚集寄主树木过程中所发挥的重要作用的概括变得非常困难。     

Metabarcoding of fungal communities associated with bark beetles

Many species of fungi are closely allied with bark beetles, including many tree pathogens, but their species richness and patterns of distribution remain largely unknown. We established a protocol for metabarcoding of fungal communities directly from total genomic DNA extracted from individual beetles, showing that the ITS3/4 primer pair selectively amplifies the fungal ITS. Using three specimens of bark beetle from different species, we assess the fungal diversity associated with these specimens and the repeatability of these estimates in PCRs conducted with different primer tags. The combined replicates produced 727 fungal Operational Taxonomic Units (OTUs) for the specimen of Hylastes ater, 435 OTUs for Tomicus piniperda, and 294 OTUs for Trypodendron lineatum, while individual PCR reactions produced on average only 229, 54, and 31 OTUs for the three specimens, respectively. Yet, communities from PCR replicates were very similar in pairwise comparisons, in particular when considering species abundance, but differed greatly among the three beetle specimens. Different primer tags or the inclusion of amplicons in separate libraries did not impact the species composition. The ITS2 sequences were identified with the Lowest Common Ancestor approach and correspond to diverse lineages of fungi, including Ophiostomaceae and Leotiomycetes widely found to be tree pathogens. We conclude that Illumina MiSeq metabarcoding reliably captures fungal diversity associated with bark beetles, although numerous PCR replicates are recommended for an exhaustive sample. Direct PCR from beetle DNA extractions provides a rapid method for future surveys of fungal species diversity and their associations with bark beetles and environmental variables.     

Catches in pitfall traps in relation to mean densities of carabid beetles

Summary Based on continuous pitfall catches summed over the whole activity period for the carabid beetles Pterostichus versicolor Sturm and Calathus melanocephalus L., a satisfactorily linear relationship is presented between the mean densities in several habitats and in different years and the numbers of beetles trapped. This means that with the help of continuous pitfall sampling a reliable relative measure of the sizes of carabid populations can be obtained.Simulation experiments show that for individuals of P. versicolor the distances covered each day should be represented by strips of some width instead of by lines to obtain numbers trapped that equal the field data. The influence on numbers trapped of the high mobility of beetles in unfavorable habitats is probably not significant. Simulation experiments with C. melanocephalus indicate that a much lower capture efficiency than for P. versicolor has to be programmed to fit the simulation results with the field data.The catch from different types of pitfalls is not always proportional to trap perimeter due to different vegetation cover of the immediate trap surroundings and to different kinds of roofs placed over the pitfalls. For the same species the capture efficiency of a pitfall can also change throughout the season.Simulation experiments show that the number and shape of the pitfalls need not influence the catches, provided that the traps keep the same total perimeter and are broadly spaced.Remodelling of some data from the literature reveals that the linear relationship between mean densities and year-catches in pitfalls is probably a general one among carabid species. This leads to the hypothesis that carabid beetles have to work through a fixed reproductive program each year by means of an almost constant total amount of locomotory activity which is realized step by step during spells of suitable weather.Some recommendations are given for intraspecific and interspecific comparisons of pitfall catches between sites and years.Communication no. 196 of the Biological Station of the Agricultural University (Wageningen), Wijster (Drenthe), The Netherlands     

Monitoring of wheat insects and their natural enemies using sticky traps in wheat

Beneficial arthropods and wheat insects were monitored using sticky traps through large-scale field in Saxony, Germany before and after insecticide applications. The tested compounds (Karate, Biscaya and NeemAzal T/S) were sprayed twice at Elongation stage (GS 32) and at the heading stage (GS 55). Monitoring was conducted for four weeks after each treatment. Cereal aphids, thrips, leafhoppers, cereal leaf beetles, cereal bugs and also many natural enemies such as predators (lady beetles, lacewings, syrphids, dance flies and spiders) and parasitoids (parasitic wasps) were surveyed. The results proved that Karate caused the highest per cent mortality to wheat insect pests. Karate also reduced natural enemy diversities. Biscaya and NeemAzal T/S is correlated with an equivalent mortality per cents to wheat insect pests and resulted in a smaller effects on natural enemies compared with Karate. Leafhoppers were less affected than Thrips and cereal bugs. Parasitoid wasps and spider were more tolerant, while lacewings and dance flies were more susceptible to insecticide effects. Finally, natural insecticides and predators and parasitoids could be highly compatible with a hygienic environment.     

Toxigenic fungi associated with stored corn

A total of 246 fungal isolates were obtained from 25 moldy corn samples collected in central Iowa. Either water or ether extracts of all corn samples except one exhibited some degree of toxicity in mice and/or ducklings. At least one toxigenic isolate was obtained from each corn sample. Extracts of cultures of 99 of the fungal isolates, involving 13 genera, produced death in one or more of the assay animals. The majority of the toxigenic isolates belonged to the generaAspergillus andPenicillium. Three isolates ofTrichothecium roseum were highly toxic to ducklings and mice. The ducklings exhibited a flaccid paralysis shortly after receiving an oral dose of extracts of these three isolates. Death frequently occurred following a second oral dose given 24 hr later. Mice given intraperitoneal injections of extracts ofT. roseum first exhibited lethargy and intermittent tremors, then hyperemotivity, roughening of the hair coat, abdominal respirations, incoordination, dyspnea and clonic convulsions with death occurring usually within 10 to 20 min.

---

Zusammenfassung Insgesamt sind 246 Pilzisolierungen von 25 verschimmelten Getreideproben, die in Zentral-Iowa gesammelt worden sind, erhalten worden. Sowohol Wasser- wie Aether-Extrakte von allen Getreideproben zeigten eine gewisse Toxizität in Mäusen als auch in Entchen außer einer einzigen Probe. Kulturextrakte von 99 der Pilzisolierungen, die 13 Gattungen umfaßten, verursachten den Tod in einem oder in mehreren Versuchstieren. Die Mehrheit der toxischen Isolierungen gehörte den GattungenAspergillus undPenicillium an. Drei Isolierungen vonTrichothecium roseum waren hoch toxisch für Entchen und Mäuse. Die Entchen zeigten eine schlaffe Paralyse kurz nachdem sie eine orale Dosis der Auszüge dieser drei Isolierungen erhalten hatten. Tod erfolgte häufig nach der zweiten oralen Dosis innerhalb 24 Stunden. Mäuse, die eine intraperitoneale Injektion der Auszüge vonT. roseum erhielten, zeigten erst eine Lethargie und einen intermittierenden Tremor, dann eine Hyperemotivität, aufgerauhte Haarbedeckung, abdominale Atmung, Inkoordination, Dyspnoe, klonische Zuckungen und Tod gewöhnlich innerhalb 10 bis 20 Minuten.

     

Ophiostomatoid fungi isolated from Japanese red pine and their relationships with bark beetles

We isolated ophiostomatoid fungi from bark beetles infesting Pinus densiflora and their galleries at 24 sites in Japan. Twenty-one ophiostomatoid fungi, including species of Ophiostoma, Grosmannia, Ceratocystiopsis, Leptographium, and Pesotum, were identified. Among these, 11 species were either newly recorded in Japan or were previously undescribed species. Some of these fungal species were isolated from several bark beetles, but other species were isolated from only a particular beetle species. Thus, it is suggested that some ophiostomatoid fungi have specific relationships with particular beetle species. In addition, fungus-beetle biplots from redundancy analysis (RDA) summarizing the effects of beetle ecological characteristics suggested that the association patterns between bark beetles and the associated fungi seemed to be related to the niches occupied by the beetles.