The encounter of distinct Morimus asper (Coleoptera: Cerambycidae) phylogeographic lineages on the Balkan Peninsula: conservation implications

Journal of Insect Conservation - The first step in monitoring and management of any taxa includes resolving taxonomic ambiguities which complicate the identification of its management units. One of...     

Adaptations to captive breeding of the longhorn beetle Morimus funereus (Coleoptera: Cerambycidae); application on amylase study

Captive breeding has been suggested as a method of conservation for many vertebrates, and is increasingly being proposed as a strategy for invertebrates. In this study, the growth, development and fertility of adults of the vulnerable cerambycid Morimus funereus reared in captivity are examined. Two oviposition cycles; from May to September and from January to March were studied and larvae from wild adults and from the progeny of captive adults (second generation larvae) were examined. Five to 12 instars were observed during larval development. Larval development was completed in 218 days (average) for the progeny of wild adults with an average mortality rate of 10.3% and in 226 days (average) for larvae from captive adults with mortality rate of 34.9%. First generation larval body weights were disparate during development, while second generation larvae had similar weights with no significant differences. In this study we have tested the potential of captive breaded M. funereus larvae as a model for investigation of digestive enzymes. Amylase from the midgut of larvae reared under laboratory conditions showed twofold higher specific activities with a decreased number of isoforms expressed, as compared to the enzyme from field-collected larvae. Captive breeding of M. funereus can be used in the future as a part of an effective conservation strategy for this rare insect species.     

Purification and properties of midgut alpha-amylase isolated from Morimus funereus (Coleoptera: Cerambycidae) larvae

Using soluble starch as a substrate five isoforms of alpha-amylase were identified in a crude extract of Morimus funereus larvae. The main alpha-amylase (termed AMF-3) was purified by gel filtration chromatography and anion exchange chromatography to obtain a single band on sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE). Its enzymatic purity was confirmed by an in-gel activity assay after SDS-PAGE. The purity of AMF-3 was increased 112-fold with a 15.4% yield. AMF-3 had apparent molecular masses of 33 and 31 kDa when analysed using SDS-PAGE and Superdex 75 FPLC gel filtration chromatography, respectively and a calculated isoelectric point of 3.2. Purified AMF-3 showed maximal activity at pH 5.2 and had an optimum activity temperature of 45 degrees C. AMF-3 retained over 90% of its maximum activity at temperatures from 45 to 60 degrees C. AMF-3 exhibited a high affinity towards soluble starch with a K(m) value of 0.43 mg/mL. Maximal AMF-3 activity was achieved in the presence of 0.1 mM CaCl(2), while at higher concentrations its activity decreased. AMF-3 activity increased with increasing NaCl concentration. AMF-3 activity was significantly inhibited by alpha-amylase wheat inhibitor. Using a number of raw starch substrates maximum AMF-3 activity was achieved with horse-radish starch, in contrast to undetectable activity towards potato starch.     

Partial purification and characterization of midgut leucyl aminopeptidase of Morimus funereus (Coleoptera: Cerambycidae) larvae

Exopeptidases of Morimus funereus larvae were partially purified and characterized. Specific leucyl aminopeptidase (LAP) activity was increased eight-fold by gel filtration of the crude midgut extract. The partially purified LAP had a molecular mass greater than 100 kDa with pH optima from 7.0-9.0 and no strict substrate specificity. M. funereus LAP preferentially hydrolyzed p-nitroanilides with hydrophobic amino acids in the active site, with a K(m) for leucine-p-nitroanilide of 0.21 mM. Zymogram analysis of an electropherogram obtained by native polyacrylamide gel electrophoresis revealed four enzymatically active proteinases using leucine-p-nitroanilide and methionine-p-nitroanilide as substrates and two enzymatically active proteinases using lysine-p-nitroanilide as a substrate. Although the optimal temperature of LAP activity was 40 degrees C, the enzyme was active over a broad temperature range from 2 to 60 degrees C. Among a number of inhibitors tested, heavy metals and 1,10-phenanthroline completely inhibited the enzyme, while methanol, ethanol and EGTA stimulated somewhat LAP activity.     

Purification and properties of major midgut leucyl aminopeptidase of Morimus funereus (Coleoptera, Cerambycidae) larvae

The major leucyl aminopeptidase (LAP) from the midgut of Morimus funereus larvae was purified and characterised. Specific LAP activity was increased 292-fold by purification of the crude midgut extract. The purified enzyme had a pH optimum of 7.5 (optimum pH range 7.0-8.5) and preferentially hydrolysed p-nitroanilides containing hydrophobic amino acids in the active site, with the highest V(max)/K(M) ratio for leucine-p-nitroanilide (LpNA). Among a number of inhibitors tested, the most efficient were 1,10-phenanthroline having a K(i) value of 0.12 mM and cysteine with K(i) value of 0.31 mM, while EGTA stimulated LAP activity. Zn(2+), Mg(2+) and Mn(2+) all showed bi-modal effects on LAP activity (activated at low concentrations and inhibited at high concentrations). The purified LAP (after gel filtration on Superose 6 column) had molecular mass of 400 kDa with an isoelectric point of 6.2. Sodium dodecylsulphate-polyacrylamide gel electrophoresis (SDS-PAGE) revealed one band of 67 kDa, suggesting that the enzyme is a hexamer. Six peptide sequences from protein band were obtained using ESI/MS-MS analysis. Comparison of the obtained peptide sequences with the EMBL-EBI sequence analysis toolbox and the BLASTP database showed a high degree of identity with other insect aminopeptidases.     

Behavioral Differences of the Insect Morimus funereus (Coleoptera,Cerambycidae) Exposed to an Extremely Low Frequency Magnetic Field

Changes in the behavior of Morimus funereus individuals were investigated as early manifestations of the contact of a living system with a changed environment primarily established via the nervous system. These experiments were aimed at revealing possible behavioral differences of a laboratory population of cerambycid beetle M. funereus in an “open field” before and after exposure to an extremely low frequency magnetic field (ELF‐MF, 50 Hz, 2 mT). The experimental groups were divided into several activity categories and exposed to ELF‐MF. The results showed that the activity increased in the groups with medium and low motor activity, but decreased in highly active individuals. High individual variability was found in the experimental groups, as well as differences in motor activities between the sexes, both before and after exposure to ELF‐MF. According to preliminary results, we assume the changes of activity in both sexes after exposure to ELF‐MF. The results showed a tendency toward locomotor activity decrease, the affect being more pronounced in females. As opposed to this type of activity, stereotypic activity of males was increased after the exposure, whereas females maintained the expected tendency of decrease. However, we did not obtain statistically significant differences because of a high individual variability and a low total number of individuals in the experiment (N = 28). Only a detailed analysis of the locomotor activity at 1‐min intervals showed some statistically significant differences in behavior between the sexes.     

星天牛幼虫龄期的划分

本文测量了人工饲养的星天牛Anoplophora chinensis(Forster)幼虫的9个形态结构数据,运用频次分布、均差分析和回归分析对测量数据进行统计分析,结果表明,星天牛幼虫可分5龄,其额宽是最佳的分龄结构,其次是上颚长和上颚宽。     

The effect of temperature on midgut and brain protein profiles in Morimus funereus larvae (Coleoptera: Cerambycidae)

The 7-days shift of M. funereus larvae, from nature to a constant temperature of 23 degrees C led to changes in midgut and brain protein quality and quantity. The changes in midgut protein profiles are characterized by an intensified protein band Mr of 29 kD, the absence of protein Mr of 22 kD and less intense bands Mr of 8.5-2.5 kD. Electrophoretic patterns of brain proteins showed less intense Mr of 66-2.5 kD protein bands.     

影响大猿叶虫交配持续时间的因素

为了探明影响大猿叶虫Colaphellus bowringi Baly交配持续时间的因素,在室内系统观察了一天中的交配起始时间、日龄、交配经历、体重、雄虫密度和温度对交配持续时间的影响.结果表明：一天中不同时间交配的成虫,其交配持续时间有显著差异,在上午8:00的交配持续时间最长,随着时间的推移,交配持续时间逐渐缩短,两者之间呈负相关.不同日龄成虫的交配持续时间有显著差异,且与成虫日龄呈明显的正相关.当雄虫与有交配经历的雌虫交配,其交配持续时间明显长于与未交配的雌虫交配,但雄虫的交配经历对交配持续时间没有影响.交配持续时间随着雌、雄体重的增加而延长,但没有显著差异.在雄虫高密度条件下的交配持续时间明显短于低密度下的交配持续时间.在22℃时成虫的交配持续时间最长,其次是25℃,28℃下最短, 温度间的交配持续时间有明显差异.这些结果表明在自然条件下大猿叶虫的交配持续时间受到许多因素的影响.     

Surprising longhorned beetle (Coleoptera, Cerambycidae) richness along an Italian alpine valley

In this paper we report about 88 longhorned beetles (Cerambycidae) species found in 6929 hectares and distributed along an altitudinal gradient of 1500 m of an Italian alpine valley (Val Genova, central-eastern Italian Alps). The species richness, result merging data from sixty years (1947-2007) of entomological surveys, corresponds to the 32% of the Italian cerambycid fauna confirming the high richness/surface ratio, probably unique in the Alps. The effect of thirteen environmental variables was tested on the species richness, but only the elevation resulted able to affect it. The species richness decrease with altitude not gradually, but experience a strong step above 1700 m a.s.l.. The highest species richness (average values of 42 species) was recorded at the lowest and mid elevations (between 800 and 1600 m a.s.l.). The species turnover along the altitudinal gradient is low suggesting moderate habitat turnover along the valley.One of the eighty-eight observed species, Tragosoma depsarium,is classified near threatened by the IUCN. Our data suggest that the wilderness of the valley close to the suitable management of grasslands and forests, help to support high level of cerambycids diversity. This biodiversity is good indicators of health of the wood saproxylic assemblages, as well an important food source for many vertebrate predators.     

Karyological notes on six beetle species from Armenia (Coleoptera: Tenebrionidae,Cerambycidae, Curculionidae)

Karyotypic details were studied in males of six beetle species from three families, viz. Tenebrionidae: Dailognatha pumila Bdy. (2n = 20, n(male) = 9 + Xy(p)), Pachyscelis musiva Ménétr. (2n = 18, n(male) = 8 + Xy(p)), Pimelia capito Kryn. (2n = 18, n(male) = 8 + Xy(p)); Cerambycidae: Agapanthia walteri Reitt. (2n = 20, n(male) = 9+Xy(p)), Agapanthia korostelevi Danilevsky (2n = 20, n(male) = 9 + Xy(p)); Curculionidae: Phyllobius caucasicus Stierl. (2n = 22, n(male) = 10 + Xy(p)). The chromosome number and sex determining system of all beetle species are described for the first time. Evolutionary trends in karyotypes of the studied beetle groups are briefly discussed.     

First record of the beetle Osphranteria coerulescens Redtenbacher, 1850 (Coleoptera,Cerambycidae) from Turkey

Abstract

The beetle Osphranteria coerulescens, until now known only from a few places in Iran and in Syria is reported from four sites in Eastern Turkey.     

A new insect cell line from the longicorn beetle Plagionotus christophi (Coleoptera: Cerambycidae)

We have established a continuous cell line from the fat body tissue of the longicorn beetle Plagionotus christophi. The cells have been serially subcultured in MGM450 medium, and the line has been designated as PC-1. The cells were grown in suspension and comprise largely flattened spindle- or oval-shaped cells morphologically related to blood cells of longicorn beetles. The chromosome number ranged from 19 to 36, with a mode of 20 (diploid). The growth curve was determined at the 100th passage, and the population doubling time was calculated to be 3.79 d. Isozyme analysis of malic enzyme, phosphoglucose isomerase, and phosphoglucomutase revealed that PC-1 cells were enzymatically distinct from coleopteran XP-1, AnCu-35, dipteran AeAl-2, and lepidopteran GaMe-LF1 cell lines.     

A method for detecting the effect of magnetic field on activity changes of neuronal populations of Morimus funereus (Coleoptera, Cerambycidae)

Modification of a new method for detecting changes in the activities of neuronal population and the nearest neuron is described. Preliminary measurements of the influence of a static magnetic field (2 mT) on neuronal population activity on eight individuals of an endangered insect species Morimus funereus are included. Five minutes exposure produced both excitatory (5/8) and inhibitory (3/8) effect on the activity of neuronal population of M. funereus antennal lobe. However, when the reversibility of induced effects was quantitatively analyzed, our results showed that they were prevailingly irreversible: (7/8) for the population, (6/8) for the nearest neuron.     

Morphology of antennal sensilla of the brown spruce longhorn beetle,Tetropium fuscum (Fabr.) (Coleoptera: Cerambycidae)

The antennal sensilla of the brown spruce longhorn beetle, Tetropium fuscum (Fabr.) (Coleoptera: Cerambycidae) were examined with particular focus on the sensilla present on the apical flagellomere. T. fuscum antennae are composed of 11 segments, namely the scape, pedicel, and nine flagellomeres. Nine types of sensilla were observed: three types of sensilla chaetica, sensilla trichodea, two types of sensilla basiconica, grooved peg sensilla, thick-walled sensilla, and Böhm bristles. Seven of these types were present on the apical flagellomere, the exceptions were sensilla chaetica type 3 and Böhm bristles. There were no significant differences in the distribution or density of sensilla present on the ninth flagellomere of males and females, except that males had significantly more sensilla chaetica type 1, which are put forward as the putative contact chemoreceptors for T. fuscum.     

光肩星天牛体内纤维素酶活性及其与寄主选择的关系（英文）

为了揭示光肩星天牛Anoplophora glabripennis Motschulsky纤维素酶与寄主选择的关系, 以4种不同寄主树种（新疆杨Populus alba var. pyramidalis Bunge、箭杆杨Populus nigra var. thevestina (Dode) Bean、合作杨Populus simonii × Populus pyramidalis cv. opera Hsu和漳河柳Salix matsudana f. lobato-glandulosa Faug et Liu来源的光肩星天牛幼虫, 和以取食5种不同树种（臭椿Ailanthus altissima、毛白杨Populus tomentosa、合作杨、旱柳Salix matsudana Koidz.和复叶槭Acer negundo Linn.）的光肩星天牛成虫为实验对象, 测定其纤维素酶活性。结果表明: 不同寄主来源的光肩星天牛幼虫内切-β-1,4-葡聚糖酶活性在1.36~2.71 μmol葡萄糖/ (g FW·h)之间, β-葡萄糖苷酶活性在2.57~4.86 μmol葡萄糖/(g FW·h)之间;取食不同树种的光肩星天牛成虫内切-β-1,4-葡聚糖酶活性在4.08~9.27 μmol葡萄糖/(g FW·h)之间, β-葡萄糖苷酶活性在2.87~6.08 μmol葡萄糖/ (g FW·h)之间。不同寄主树种来源的光肩星天牛幼虫体内纤维素酶活性无显著性差异, 取食与否以及取食树种的不同对光肩星天牛成虫纤维素酶活有较大影响。     

Different effects of coarse woody material on the species diversity of three saproxylic beetle families (Cerambycidae, Melandryidae, and Curculionidae)

The relationships between coarse woody material (CWM) and the diversity of three saproxylic beetle families (Cerambycidae, Melandryidae, and Curculionidae) were investigated. These three families responded differently toward the quality and quantity of CWM. The species richness of cerambycid beetles increased in forests with more CWM in the early stage of decomposition. The richness of saproxylic curculionids increased in stands with more tree species. No clear trend was evident for melandryid diversity. The CWM differed in quality and quantity among three forest types [middle-aged larch (Larix kaempferi [Lamb.] Carrière) plantation, secondary forest, and old-growth forest] and between two forest-management practices (thinning and long rotation). Forest type and management practices affected CWM and the number of tree species, and CWM and the number of tree species influenced the diversity of saproxylic families differently. Because each decomposition stage is considered to be important for saproxylic beetles at the species level, a constant supply of CWM is necessary to maintain a suitable balance of CWM through these decomposition stages in forests to maintain the diversity of saproxylic beetles.     

Evidence for limited spatial spread in an exotic longhorn beetle, Tetropium fuscum (Coleoptera: Cerambycidae)

The longhorn beetle Tetropium fuscum F. (Coleoptera: Cerambycidae) has become established in Nova Scotia, Canada, where it coexists with Tetropium cinnamopterum Kirby. The two Tetropium species share a similar ecological niche and use the same volatile cues for mate attraction. Exotic T. fuscum was introduced near Halifax, Nova Scotia, in approximately 1990, but the rate of its spread 20 yr later has not been documented. We report a large-scale, 3-yr study that investigates the distribution of T. fuscum relative to its site of introduction. Traps baited with male-produced pheromone and host volatiles were used to estimate the relative abundance of the two Tetropium species. Adult T. fuscum emerged 1-2 wk earlier than T. cinnamopterum each year between 2008 and 2010. The spatial distribution of T. fuscum was characterized by a sharp decline in abundance in relation to its point of introduction, up to a threshold distance of approximately 80 km beyond which T. fuscum is rare in comparison with native T. cinnamopterum. The restricted range of T. fuscum 20 yr after its introduction may be attributed to limited dispersal of adults or reproductive failures of low-density populations. The distribution of T. fuscum seemed stable between 2008 and 2010. In 1 of 3 yr, the abundance of T. cinnamopterum increased with the distance to the site of introduction of T. fuscum, which suggests competitive interactions between the two Tetropium species.     

栗山天牛幼虫龄数和龄期的测定

栗山天牛Massicus raddei (Blessig)是危害我国东北柞树天然林的重要蛀干害虫, 长期营隐蔽性生活, 3年发生1代, 防治困难。幼虫的龄数和龄期测定是害虫预测预报以及制订其科学治理策略的重要依据。本研究于2008-2011年通过林间定期解剖受害树进行取样, 在辽宁省宽甸县采集不同发育阶段的栗山天牛幼虫, 分别测量幼虫上颚长、 主单眼间距、 前胸背板宽、 中胸气门长和体长等5项形态指标, 利用频次分析的统计方法, 测定了栗山天牛幼虫的龄数。结果表明: 栗山天牛幼虫有6龄, 雌雄性幼虫龄数相同。上颚长、 主单眼间距和前胸背板宽3项指标均可用于分龄, 中胸气门长和体长变异较大, 不宜用作幼虫龄数划分。利用种群众数龄期法计算1-6龄幼虫的平均龄期分别为9.25, 266.85, 48.09, 51.29, 260.33和385.71 d, 幼虫期共1 021.52 d。在我国东北地区, 自然条件下栗山天牛世代发生非常整齐而且高度同步, 完成1代发育需跨越4个年份, 幼虫经历3次越冬, 第1年以2-3龄幼虫越冬, 第2年主要以4-5龄幼虫越冬, 第3年全部以末龄幼虫越冬。研究结果进一步明确了栗山天牛幼虫期的生物学特性, 为生产上合理防治该害虫提供了参考依据。     

Life history of the Asian longhorn beetle Anoplophora glabripennis (Coleoptera Cerambycidae) in southern Europe

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