Nondestructive DNA sampling from bumblebee faeces

Genetic studies provide valuable data to inform conservation strategies for species with small or declining populations. In these circumstances, obtaining DNA samples without harming the study organisms is highly desirable. Excrements are increasingly being used as a source of DNA in such studies, but such approaches have rarely been applied to arthropods. Bumblebees are ecologically and economically important as pollinators; however, some species have recently suffered severe declines and range contractions across much of Western Europe and North America. We investigated whether bumblebee faeces could be used for the extraction of DNA suitable for genotyping using microsatellite markers. We found that DNA could be extracted using a Chelex method from faecal samples collected either in microcapillary tubes or on filter paper, directly from captured individuals. Our results show that genotypes scored from faecal samples are identical to those from tissue samples. This study describes a reliable, consistent and efficient noninvasive method of obtaining DNA from bumblebees for use in population genetic studies. This approach should prove particularly useful in breeding and conservation programs for bumblebees and may be broadly applicable across insect taxa.     

Arthropod biodiversity loss and the transformation of a tropical agro-ecosystem

The coffee (Coffea arabica) agro-ecosystem in the Central Valley of Costa Rica was formerly characterized by a high vegetational diversity. This complex system has been undergoing a major transformation to capital-intensive monocultural plantations where all shade trees are eliminated. In this study we examined the pattern of arthropod biodiversity loss associated with this transformation. Canopy arthropods were sampled in three coffee farms: a traditional plantation with many species of shade trees, a moderately shaded plantation with only Erythrina poeppigeana and coffee, and a coffee monoculture. An insecticidal fogging technique was used to sample both canopy and coffee arthropods. Data are presented on three major taxonomic groups: Coleoptera, non-formicid Hymenoptera, and Formicidae. Data demonstrate that the transformation of the coffee agro-ecosystem results in a significant loss of biological diversity of both canopy arthropods as well as arthropods living in coffee bushes. Percentage of species overlap was very small for all comparisons. Furthermore, species' richness on a per tree basis was found to be within the same order of magnitude as that reported for trees in tropical forests. If results presented here are generalizable, this means that conservation efforts to preserve biological diversity should also include traditional agro-ecosystems as conservation units.     

Seasonal fluctuation, phenology and turnover of chafer assemblages – insight to the structural plasticity of insect communities in tropical farmlands

1 Studies on chafer assemblages were conducted on two farmland sites in the Terai lowland of Nepal (200 m above sea level) using light traps. During the course of a 2-year field monitoring program, a total of 4503 specimens was captured and an unexpectedly high number of syntopically co-occurring species was found: 52 from Gunganagar (GN) and 36 from Gaindakot (GK), respectively. Highest species abundances and species numbers were found during April and May.
2 Species occurrence was strongly correlated with air temperature and the maximum soil temperature, at least during the pre-monsoon season. However, assemblage structure from the two sites showed significant qualitative and quantitative changes seasonally, as well as from 1 year to the next. Turnover rates between adjacent months were in the range 26–62% (GN) and 37–70% (GK), whereas the turnover from 2004 to 2005 was 25.8% (GN) and 21.4% (GK) respectively.
3 When only dominant and subdominant taxa are considered, the seasonal change in species composition was even more striking.
4 Strong fluctuation in chafer assemblage over time suggests: (i) a possible influence of patchy habitat types and soil working on seasonal assemblage structure and (ii) colonization of suitable habitats (fields) in great part by chance.     

Non-lethal sampling of DNA from bumble bees for conservation genetics

Summary Non-lethal sampling of DNA from individuals in wild populations will often be required for studies of the conservation genetics of social insects, since it avoids destroying members of scarce or declining species. We investigated the effectiveness and consequences of methods of non-lethal sampling of DNA from bumble bee workers. In an experiment with two captive and confined Bombus terrestris colonies, we found that, unlike sampling haemolymph, sampling the terminal portion of the tarsus of a mid-leg of a worker reliably yielded amplifiable microsatellite DNA and did not significantly reduce worker survivorship. In a further experiment with four B. terrestris colonies whose workers were allowed to forage freely at flowers in the external environment, tarsal sampling of either a mid-leg or a hind-leg had no significant effects on worker survivorship, the mean body mass of foraging workers, the frequency or duration of foraging trips, mass of pollen loads or mass of nectar loads. We therefore suggest that tarsal sampling of either a mid-leg or a hind-leg is an effective and acceptable means of non-lethally sampling DNA from workers in wild populations of bumble bees, because effects on individual and colony performance are likely to be absent or minimal.Received 2 December 2002; revised 10 April 2003; accepted 25 April 2003.     

Noninvasive sampling of DNA from larval exuvia in diving beetles of the genera Cybister and Dytiscus

Nonlethal DNA sampling is a highly recommendable method in molecular genetic studies of protected and endangered species. To develop a demonstrably nonlethal method of obtaining DNA from larvae of endangered diving beetles (Cybister brevis, C. lewisianus, C. limbatus, C. rugosus, Dytiscus sharpi sharpi and D. sharpi validus), we obtained the larval exuvia (molted skin) of these endangered diving beetles under laboratory conditions. A single exuvia 24 h after molting was sufficient to allow polymerase chain reaction (PCR) detection of a mitochondrial DNA gene, cytochrome‐c oxidase subunit I (COI), and the sequence of the COI gene could be determined directly. Sequences obtained from the exuvial samples were used to further find similarities within DDBJ/EMBL/GenBank. Genomic DNA from the samples was successfully isolated, and we identified the species. This process suggests that exuvia provides a good sample for extracting DNA from endangered diving beetle larvae without killing them.     

Polymorphic microsatellite DNA markers from the Oriental Fruit Fly Bactrocera dorsalis (Hendel)

Six polymorphic microsatellite loci are isolated from the Oriental fruit fly Bactrocera dorsalis (Hendel), an agricultural pest in Asia, including Taiwan. To assess their potential utility as high‐resolution genetic markers, polymerase chain reaction (PCR) primers, amplification conditions, and an automated fluorescence detection protocol were developed. In analyses of 71 individual flies from six different areas of Taiwan, allele numbers ranged from five to 25 were detected for each locus. The observed heterozygosity ranged between 0.268 and 0.737 among these loci. No linkage disequilibrium was found. These microsatellite markers have potential utility to population structure and gene flow studies of B. dorsalis (Hendel).     

Isolation of microsatellite DNA and analysis on genetic diversity of endangered fish,Hucho taimen (Pallas)

Hucho taimen (Pallas) is an endangered species in China. To evaluate the species’ genetic diversity and population genetic structure, we isolated six polymorphic microsatellite markers from its genomic libraries by (ACA)₁₆ enriched, and characterized using 61 wild individuals. The number of alleles per locus ranged from two to seven, expected heterozygosity ranged from 0.3200 to 0.7410, polymorphism information content ranged from 0.3047 to 0.6896. These markers will be useful for the genetic variation assessment of taimen.     

New species of scarabaeoid beetles from the family Glaresidae (Coleoptera: Scarabaeoidea) from the Cretaceous of Transbaikalia

A new species from the family Glaresidae, Cretoglaresis ovalis, sp. nov., is described from the Lower Cretaceous site Baisa in Transbaikalia. Improved diagnosis of the genus Cretoglaresis Nikolajev, 2007 is given.     

A revision of the South African Chaetocnema gahani speciesgroup,with descriptions of four new flea beetle species (Coleoptera: Chrysomelidae)

The Chaetocnema gahani species-group from Southern Africa is herein analyzed and revised. It includes C. gahani Jacoby 1897, C. brincki (Bechyné 1959), C. capeneri n. sp., C. danielssoni n. sp., C. sudafricana n. sp., and C. tuckeri n. sp. A key to all the species is presented. Line drawings of male and female genitalia and scanning electron micrographs of six diagnostic morphological characters are also included, along with ecological and zoogeographical data. Finally, discriminant analyses using the six morphological characters supports the close affinity of the six species within the C. gahani group.     

A review of Turkish saproxylic beetles from the European Red List

The conservation of beetles that depend on dying or dead wood (i.e. saproxylic), has received a great deal of attention in many parts of the world in recent years. Human activities such as urbanization and logging, and their results, e.g. global warming, destroy natural ecosystems and threaten unusual species such as Lucanus cervus (L. 1758), Rosalia alpina (L. 1758), Limoniscus violaceus (Müller 1821) and Osmoderma eremita (Scopoli 1763). Attempts have been made to conserve the habitats upon which these species depend. Although represented by a large number of species, saproxylic beetles in Turkey have not received much attention. Although some Turkish studies have acknowledged their value, comprehensive investigations of these species are very limited. The aim of this study is to make a list, from the literature, of the species that are endangered in Europe and found in Turkey, to highlight the importance of these species. The research includes 151 saproxylic beetle species belonging to the families Bostrichidae, Cerambycidae, Elateridae, Eucnemidae, Scarabaeidae, Euchiridae, Lucanidae and Mycetophagidae. The list of Turkish saproxylic beetles was prepared using the European Red List of Saproxylic Beetles. Information is provided about the IUCN Red List category in Europe, the locations of the species in Turkey, and their global geographic range. We gathered all data about these species from the international literature. Furthermore, we also include in the list certain species whose locations are not specified, but which inhabit Turkey. We obtained their records from some catalogs in Turkey. In addition to the list, we prepared three distribution maps for Turkey. The maps show distribution in Turkey of some species in the Endangered, Vulnerable and Near Threatened categories in the European Red List. It is expected that this study will lead to comprehensive studies on saproxylic beetles from Turkey.     

Identification of insect pests as food foreign materials using DNA barcodes

Insect foreign materials in food are of great economic and hygienic significance. However, identifying these species with any certainty requires an expert taxonomist and can be a time consuming process. Furthermore, insects are found as body parts or they are immature specimens that cannot be identified by conventional means. For these reasons, a reference database and efficient means of identification by non‐specialists are necessary to control insect pests. In this study, we chose 15 important insect pest species, because they had a higher probability of being included in human foods. We tested the utility of the cytochrome c oxidase I (COI) DNA barcodes for the identification. A 658‐bp fragment of the COI gene was sequenced, aligned, and a sequence data bank was constructed. As a result, the COI barcode sequence was suitable for identifying insect pests as food foreign materials. Photographs of morphological key characters by stereoscopic microscope and a pictorial key of the species are provided.     

蜜蜂EST中的微卫星分析

为加速分子标记在蜜蜂遗传、进化与行为等方面的利用,分析了简单重复序列(Simple Sequence Repeats,SSRs)在蜜蜂EST中的分布频率与密度。所分析的蜜蜂EST数据集包含15869条序列,总长为7．9Mb。结果显示,蜜蜂ESTs中SSRs的频率为1／0．52kb,其中6碱基重复基序占总SSRs的45．0％,是最丰富的重复单元,而2、1、3、4与5碱基重复基序分别占总SSRs的17．9％、14．1％、11．6％、9．2％和2．2％。同时,在各种SSRs重复单元中,富含A碱基的重复单元占据优势地位,如：A、AT、AG、AC、AAT、AAG、AAC、AAAT、AAAG、AAAAG、AAAAT、AATAT、AAAAAG和AAAAAT重复基序,而富含G碱基的重复单元在基因编码区中含量较低。进一步分析显示：蜜蜂SSRs在冗余与非冗余EST数据集中的分布频率与密度相似,仅存在极小的偏差,表明可从现有的部分ESTs数据中方便地获取有效的微卫星标记。     

前裂长管茧蜂寄生行为对桔小实蝇幼虫生理指标的影响

通过观察测定了桔小实蝇幼虫生长发育过程中血淋巴蛋白种类和血细胞的变化以及前裂长管茧蜂的寄生行为对桔小实蝇幼虫各项生理指标的影响。结果表明:不同日龄桔小实蝇幼虫的血细胞浓度随着虫龄的增加呈显著上升趋势,由2龄的16.53×106cells/mL,到3龄的30.14×106cells/mL直至蛹前期的35.94×106cells/mL,但血淋巴蛋白种类没有明显变化。和未寄生幼虫相比,寄生后的幼虫各类型血细胞的浓度均下降,但差异均不显著;血淋巴蛋白种类无明显增减,但浓度有所变化;寄生4 h后血淋巴蛋白质浓度显著降低,接近22 h时升高,至化蛹前期浓度再次下降;寄生行为使幼虫的发育历期从8 d延长至9～11d;4日龄幼虫在被寄生后的第4 d起体重显著高于未被寄生的桔小实蝇幼虫。     

New jewel beetles (Coleoptera: Buprestidae) and similar beetles from the Cretaceous and Early Paleogene of Asia

A new genus and two new species of jewel beetles are described, Andakhudukia ponomarenkoi gen. et sp. nov. from the Lower Cretaceous of Mongolia and Metabuprestium ustkivdense sp. nov. from the lowermost Paleogene of the Amur Region. In addition, four new monotypic genera that share some features with jewel beetles are described: Cretoelaterium kazanovense gen. et sp. nov. from the Lower Cretaceous of Eastern Transbaikalia and Cretopoena gratshevi gen. et sp. nov. from the Lower Cretaceous of Mongolia have been referred to the families Elateridae and Eucnemidae; Cretofalselaterium baiankhongoricum gen. et sp. nov. from the Lower Cretaceous of Mongolia and Cretogermen turonicum gen. et sp. nov. from the Upper Cretaceous of Kazakhstan have been erected for isolated elytra and placed in Coleoptera incertae sedis.     

Description of Hydrobius orientalis sp. n., the first species of the subtribe Hydrobiusina from the Oriental Region (Coleoptera: Hydrophilidae: Hydrophilini)

The first Oriental species of the water scavenger beetle subtribe Hydrobiusina, Hydrobius orientalis sp. n., is described and illustrated from Guandong Province, China. It is unique in the subtribe by lacking elytral serial punctures. The species appears most closely related to Hydrobius melaenus (Germar, 1824) of eastern North America.     

Effects of environmental factors on the species composition of aquatic insects in irrigation ponds

Although irrigation ponds contribute to the conservation of aquatic biodiversity, they have experienced declines in recent years. We therefore examined the relationships between various environmental factors and the community composition of aquatic insects, specifically insect predators, in irrigation ponds to gain knowledge that would aid in the conservation and restoration of biodiversity. We selected Odonata, Hemiptera and Coleoptera as target taxonomic groups and conducted censuses of these groups in 21 ponds in Shiga, central Japan. In total, we collected 30 and 10 species (or species groups) of Odonata and Hemiptera, respectively, and 17 species of Coleoptera. A partial canonical correspondence analysis revealed that the following four environmental factors significantly affected the species composition of aquatic insect communities: the number of emergent plant species, percent concrete revetment, presence of litter and peripheral length. Among these variables, the number of emergent plant species was the most potent factor, perhaps because emergent plants serve as sites for oviposition and emergence, and provide refugia for aquatic insects (odonate nymphs in particular). In contrast, some species specifically inhabited sites poor in emergent plants. This study shows that reductions in concrete revetments are necessary for the conservation of biodiversity. This would lead to increases in the number of aquatic plant species, which provide habitats and oviposition sites for many aquatic insects. Furthermore, to enrich the local biodiversity of aquatic insects, groups of irrigation ponds with different environments are needed.     

植物总DNA样品的快速制备

利用Ｑｉａｇｅｎ微量植物ＤＮＡ提取试剂盒,在１小时内即可从植物组织获得总ＤＮＡ,提取过程中勿需酚／氯仿和ＳＤＳ抽提,操作简便、快捷。所得ＤＮＡ样品的ＯＤ２６０／ＯＤ２８０值在１．７－１．９之间。样品纯度高;该样品不含ＰＣＲ反应抑制剂及其他酶反应抑制剂,可被各种限制性内酶完全降解,适合于ＰＣＲ、印迹、ＲＡＰＤ、ＡＦＬＰ和ＲＦＬＰ分析等各种下游应用。     

Isolation and characterization of microsatellite DNA markers from mangrove crab,Scylla paramamosain

The first five variable microsatellite DNA loci for mangrove crab, Scylla paramanosain, were developed. Allelic variation and other characteristics at these loci were examined in this species captured at the Urado Bay, Japan. The number of alleles per locus ranged from 24 to 44. The expected heterozygosity across loci ranged from 0.900 to 0.999 and probability of identity (P_I) ranged from 2.8 × 10^?3 to 1.7 × 10^?2. Therefore, these microsatellite markers could be useful for estimating effect of stock enhancement release and population genetic structure of mangrove crab.     

Twenty-one novel microsatellite DNA loci isolated from the Puget Sound white-crowned sparrow, Zonotrichia leucophrys pugetensis

The white‐crowned sparrow, Zonotrichia leucophrys, has served as a model species for studies of song and reproductive physiology. Here, we describe primers for 21 novel microsatellite loci isolated from the Puget Sound subspecies, Zonotrichia leucophrys pugetensis, which will be useful for parentage and population genetic analyses. Based on genotypes from seven to 22 adult birds from one population, the average number of alleles per locus was 10.9 (four to 21 alleles) and observed heterozygosity varied from 0.50 to 1.00. All loci also amplified products in at least one of three other passerine species tested.