Evaluating Drosophila suzukii immunomarking for mark‐capture research

Drosophila suzukii Matsumura (Diptera: Drosophilidae) utilizes ‘Himalaya’ blackberry, Rubus armeniacus Focke (Rosaceae), as a host and may invade berry and stone fruit crops from field margins containing this invasive weed. Laboratory and semi‐field studies were conducted to determine (1) the persistence of protein marks including 10% chicken egg whites (egg albumin protein), 20% bovine milk (milk casein protein), and 20% soy milk (soy trypsin inhibitor protein) on topically sprayed D. suzukii, (2) protein retention on blackberry leaves, and (3) D. suzukii acquisition of protein after exposure to marked blackberry leaves for up to 14 days after application. All flies and leaves were assayed for the presence of the protein marks using protein‐specific enzyme‐linked immunosorbent assays. Egg albumin, milk casein, and soy trypsin proteins persisted on 94, 49, and 25% of the topically marked D. suzukii, respectively, throughout the 14‐day study period. Egg albumin was retained on 100% of treated leaves for 14 days, regardless of environmental conditions. At least 50% of flies exposed residually to egg albumin‐treated leaves were marked for 3 days, regardless of exposure time and environmental conditions. However, increasing fly exposure time to treated leaves in April and June appeared to improve protein mark acquisition. Acquisition of protein by flies from treated leaves for milk casein was inconsistent, and poor for soy trypsin, despite detectable levels on treated leaves. Egg albumin had the longest and most consistent persistence on flies, leaves, and flies exposed to leaves in laboratory and semi‐field studies, under a variety of environmental conditions and exposure times.     

A protein‐based approach to mark arthropods for mark‐capture type research

A series of studies was conducted to test methods for marking a wide variety of arthropods with inexpensive proteins for mark‐capture dispersal research. The markers tested included egg albumin protein in chicken egg whites and casein protein in bovine milk. The first study qualified the effectiveness of the two marks on more than 50 arthropod species inhabiting cotton via two application procedures. The application methods included: (1) a topical plus residue protein application, and (2) a residue‐only protein application. Both protein marks, regardless of the method of application, were readily retained on the arthropod assemblage over the duration of the study. The second study determined how rapidly insects acquire chicken egg albumin protein after contact exposure to cotton tissue sprayed with an egg whites solution. Under laboratory conditions, the vast majority of adult Hippodamia convergens Guérin‐Méneville (Coleoptera: Coccinellidae) and Lygus hesperus Knight (Heteroptera: Miridae) acquired the mark after 5 min, and immature Trichoplusia ni (Hübner) (Lepidoptera: Noctuidae) acquired the marker after 40 min. The third study determined how rapidly H. convergens and L. hesperus acquire bovine casein protein after contact exposure to either alfalfa, Medicago sativa L. (Fabaceae), or lesquerella, Lesquerella fendleri (Watson) (Brassicaceae), plants sprayed with a bovine milk solution. These insects rapidly acquired the casein mark from the plant residue under field conditions. A final study determined how long H. convergens retain casein protein after 24‐h exposure to alfalfa and lesquerella plants containing a 7‐day‐old residue of bovine milk. Approximately 95% of the H. convergens maintained the casein mark for 2 days after removal from each type of plant.     

Development of a standardized protein immunomarking protocol for insect mark–capture dispersal research

A field study was conducted to test the marking efficiency of broadcast spray applications of protein marks on stationary (represented by cadavers) and free‐roaming lady beetles Hippodamia convergens Guérin‐Méneville that were strategically placed in blooming alfalfa plots. The marks tested included three different concentrations of egg albumin from chicken egg white, casein from bovine milk and trypsin inhibitor from soy milk. The cadaver and free‐roaming beetle treatments served to measure the acquisition and retention of each protein treatment regime by direct contact with the spray solution and by residual contact with protein‐marked residue on alfalfa, respectively. In addition, the vertical distribution of marking efficacy was determined by sampling alfalfa plant tissue and beetle cadavers that were located on the upper and lower portion of the plant canopy. The data indicated that the backpack spray apparatus was very effective at uniformly administering the various protein marks, regardless of the concentration, throughout the entire plant canopy. Also, the free‐roaming beetles readily self‐marked by contact exposure to protein‐treated plants. We also identified concentrations of each protein type that will mark about 90% of the resident beetle population. Moreover, if a mark–capture‐type study only requires two unique protein marks, we determined that concentrations of 25% for egg white and 100% for bovine milk could be used to mark 98% of the population. Our results provide a significant step towards standardizing protein immunomarking protocols for insect mark–capture dispersal research. In addition, we identify several areas of research that are needed to further standardize the protein mark–capture procedure.     

Distribution and activity of Drosophila suzukii in cultivated raspberry and surrounding vegetation

Spotted wing drosophila, Drosophila suzukii (Matsumura) (Diptera: Drosophilidae), may utilize wild ‘Himalaya’ blackberry (HB) Rubus armeniacus Focke or other non‐crop plants as refugia and possibly exploit adjacent field margins before colonizing cultivated fruiting crops. Studies were conducted to determine the role of field margins containing HB and their effect on D. suzukii activity, density and distribution in an adjacent commercial red raspberry crop. One‐ha plots adjacent to field margins containing HB or known non‐host (NH) grass crops were established in 2011 and 2012 and replicated three times. Each plot contained two transects with monitoring traps for D. suzukii in the field margin (0 m) and spaced approximately 10 (crop boundary), 40, 70 and 100 m into the adjacent crop (n = 10 traps/plot). Field margin vegetation was treated with a 10% chicken egg white mark solution weekly from pre‐harvest until the end of harvest using a cannon sprayer. Adult D. suzukii were collected from traps weekly and analysed for the presence of the egg white mark using an egg white‐specific enzyme‐linked immunosorbent assay (ELISA). During both years, marked flies and total flies were captured in higher numbers in HB field margins, whereas virtually no flies were captured in field margins containing no known alternative host. Similarly, more flies were captured in the crop near HB than near NH. Spatial Analysis by Distance IndicEs (SADIE) and mean D. suzukii trap captures additionally displayed significantly higher fly densities in the raspberry field near HB than near NH. These results suggest that HB may contribute to elevated D. suzukii populations and pest pressure in comparison with field margins containing no known alternate host vegetation for D. suzukii. Having closely adjacent non‐crop alternate host landscapes may result in increased D. suzukii pest pressure.     

Coccinellid interactions mediated by vegetation heterogeneity

Environmental heterogeneity can have profound effects on agroecosystem function and it is important for improving ecosystem services such as biological control. Promoting system diversity via non‐crop plants is one method for increasing habitat heterogeneity within farmscapes. Non‐crop plants provide access to refuges and alternative food resources provide multiple benefits to enhance populations of arthropod predators. In this study, we examined the effects of small‐scale spatial structure on life‐stage specific interactions between the native coccinellid, Hippodamia convergensGuérin‐Méneville, and the exotic Harmonia axyridis (Pallas) (both Coleoptera: Coccinellidae), which overlap in spatial distribution in many crop systems. Squash [Cucurbita pepo L. (Cucurbitaceae)] and non‐crop mugwort [Artemisia vulgaris L. (Asteraceae)] plants with and without aphids were used as a model of spatial heterogeneity in micro‐ and mesocosm experiments. In response to factorial treatment combinations, we evaluated oviposition behavior, egg predation, larval survival, and larval predator‐prey and predator‐predator interactions. Adult H. convergens displayed higher foraging activity on aphids when exposed to complex habitats containing a non‐crop plant. In the presence of the exotic coccinellid, H. convergens preferred to deposit eggs on the non‐crop plant. Furthermore, a combination of spatial heterogeneity and prey availability reduced larval intraguild predation and cannibalism, and improved reproductive output of H. convergens by reducing intra‐ and interspecific egg predation. Our results provide evidence that life‐stage‐specific intraguild interactions are mediated by access to non‐crop plants. Thus, the introduction or maintenance of non‐crop plants has the potential to enhance coexistence of multiple natural enemies and improve top‐down control of pests.     

Cofeeding intra‐ and interspecific transmission of an emerging insect‐borne rickettsial pathogen

Cat fleas (Ctenocephalides felis) are known as the primary vector and reservoir of Rickettsia felis, the causative agent of flea‐borne spotted fever; however, field surveys regularly report molecular detection of this infectious agent from other blood‐feeding arthropods. The presence of R. felis in additional arthropods may be the result of chance consumption of an infectious bloodmeal, but isolation of viable rickettsiae circulating in the blood of suspected vertebrate reservoirs has not been demonstrated. Successful transmission of pathogens between actively blood‐feeding arthropods in the absence of a disseminated vertebrate infection has been verified, referred to as cofeeding transmission. Therefore, the principal route from systemically infected vertebrates to uninfected arthropods may not be applicable to the R. felis transmission cycle. Here, we show both intra‐ and interspecific transmission of R. felis between cofeeding arthropods on a vertebrate host. Analyses revealed that infected cat fleas transmitted R. felis to naïve cat fleas and rat fleas (Xenopsylla cheopis) via fleabite on a nonrickettsemic vertebrate host. Also, cat fleas infected by cofeeding were infectious to newly emerged uninfected cat fleas in an artificial system. Furthermore, we utilized a stochastic model to demonstrate that cofeeding is sufficient to explain the enzootic spread of R. felis amongst populations of the biological vector. Our results implicate cat fleas in the spread of R. felis amongst different vectors, and the demonstration of cofeeding transmission of R. felis through a vertebrate host represents a novel transmission paradigm for insect‐borne Rickettsia and furthers our understanding of this emerging rickettsiosis.     

Egg load dynamics and the risk of egg and time limitation experienced by an aphid parasitoid in the field

Insect parasitoids and herbivores must balance the risk of egg limitation and time limitation in order to maximize reproductive success. Egg and time limitation are mediated by oviposition and egg maturation rates as well as by starvation risk and other determinants of adult lifespan. Here, we assessed egg load and nutritional state in the soybean aphid parasitoid Binodoxys communis under field conditions to estimate its risk of becoming either egg‐ or time‐limited. The majority of female B. communis showed no signs of egg limitation. Experimental field manipulations of B. communis females suggested that an average of 4–8 eggs were matured per hour over the course of a day. Regardless, egg loads remained constant over the course of the day at approximately 80 eggs, suggesting that egg maturation compensates for oviposition. This is the first case of such “egg load buffering” documented for a parasitoid in the field. Despite this buffering, egg loads dropped slightly with increasing host (aphid) density. This suggests that egg limitation could occur at very high host densities as experienced in outbreak years in some locations in the Midwestern USA. Biochemical analyses of sugar profiles showed that parasitoids fed upon sugar in the field at a remarkably high rate. Time limitation through starvation thus seems to be very low and aphid honeydew is most likely a source of dietary sugar for these parasitoids. This latter supposition is supported by the fact that body sugar levels increase with host (aphid) density. Together, these results suggest that fecundity of B. communis benefits from both dynamic egg maturation strategies and sugar‐feeding.     

Phylogeography and population genetics of pine butterflies: Sky islands increase genetic divergence

The sky islands of southeastern Arizona (AZ) mark a major transition zone between tropical and temperate biota and are considered a neglected biodiversity hotspot. Dispersal ability and host plant specificity are thought to impact the history and diversity of insect populations across the sky islands. We aimed to investigate the population structure and phylogeography of two pine‐feeding pierid butterflies, the pine white (Neophasia menapia) and the Mexican pine white (Neophasia terlooii), restricted to these “islands” at this transition zone. Given their dependence on pines as the larval hosts, we hypothesized that habitat connectivity affects population structure and is at least in part responsible for their allopatry. We sampled DNA from freshly collected butterflies from 17 sites in the sky islands and adjacent high‐elevation habitats and sequenced these samples using ddRADSeq. Up to 15,399 SNPs were discovered and analyzed in population genetic and phylogenetic contexts with Stacks and pyRAD pipelines. Low genetic differentiation in N. menapia suggests that it is panmictic. Conversely, there is strong evidence for population structure within N. terlooii. Each sky island likely contains a population of N. terlooii, and clustering is hierarchical, with populations on proximal mountains being more related to each other. The N. menapia habitat, which is largely contiguous, facilitates panmixia, while the N. terlooii habitat, restricted to the higher elevations on each sky island, creates distinct population structure. Phylogenetic results corroborate those from population genetic analyses. The historical climate‐driven fluxes in forest habitat connectivity have implications for understanding the biodiversity of fragmented habitats.     

Concordance of bacterial communities of two tick species and blood of their shared rodent host

High‐throughput sequencing is revealing that most macro‐organisms house diverse microbial communities. Of particular interest are disease vectors whose microbiome could potentially affect pathogen transmission and vector competence. We investigated bacterial community composition and diversity of the ticks Dermacentor variabilis (n = 68) and Ixodes scapularis (n = 15) and blood of their shared rodent host, Peromyscus leucopus (n = 45) to quantify bacterial diversity and concordance. The 16S rRNA gene was amplified from genomic DNA from field‐collected tick and rodent blood samples, and 454 pyrosequencing was used to elucidate their bacterial communities. After quality control, over 300 000 sequences were obtained and classified into 118 operational taxonomic units (OTUs, clustered at 97% similarity). Analysis of rarefied communities revealed that the most abundant OTUs were tick species‐specific endosymbionts, Francisella and Rickettsia, and the commonly flea‐associated bacterium Bartonella in rodent blood. An Arsenophonus and additional Francisella endosymbiont were also present in D. variabilis samples. Rickettsia was found in both tick species but not in rodent blood, suggesting that it is not transmitted during feeding. Bartonella was present in larvae and nymphs of both tick species, even those scored as unengorged. Relatively, few OTUs (e.g. Bartonella, Lactobacillus) were found in all sample types. Overall, bacterial communities from each sample type were significantly different and highly structured, independent of their dominant OTUs. Our results point to complex microbial assemblages inhabiting ticks and host blood including infectious agents, tick‐specific endosymbionts and environmental bacteria that could potentially affect arthropod‐vectored disease dynamics.     

An ecological genetic delineation of local seed‐source provenance for ecological restoration

An increasingly important practical application of the analysis of spatial genetic structure within plant species is to help define the extent of local provenance seed collection zones that minimize negative impacts in ecological restoration programs. Here, we derive seed sourcing guidelines from a novel range‐wide assessment of spatial genetic structure of 24 populations of Banksia menziesii (Proteaceae), a widely distributed Western Australian tree of significance in local ecological restoration programs. An analysis of molecular variance (AMOVA) of 100 amplified fragment length polymorphism (AFLP) markers revealed significant genetic differentiation among populations (Φ_PT = 0.18). Pairwise population genetic dissimilarity was correlated with geographic distance, but not environmental distance derived from 15 climate variables, suggesting overall neutrality of these markers with regard to these climate variables. Nevertheless, Bayesian outlier analysis identified four markers potentially under selection, although these were not correlated with the climate variables. We calculated a global R‐statistic using analysis of similarities (ANOSIM) to test the statistical significance of population differentiation and to infer a threshold seed collection zone distance of ~60 km (all markers) and 100 km (outlier markers) when genetic distance was regressed against geographic distance. Population pairs separated by >60 km were, on average, twice as likely to be significantly genetically differentiated than population pairs separated by <60 km, suggesting that habitat‐matched sites within a 30‐km radius around a restoration site genetically defines a local provenance seed collection zone for B. menziesii. Our approach is a novel probability‐based practical solution for the delineation of a local seed collection zone to minimize negative genetic impacts in ecological restoration.     

Occurrence and impact of Agrilus spp. and associated egg parasitoids in hazel groves of Northwest Italy

In several hazel (Corylus avellana L.) groves in the area of Langhe (Piedmont, Northwest Italy), many hazel branches suddenly withered, and in some cases, the whole tree died, with heavy economic losses for the farmers. Symptoms of jewel beetle attacks were detected on the trees. Eight Agrilus species were caught by traps from 2007 to 2009 in the surveyed hazel groves; among them, only four species have been known to develop on hazel. On the traps, Agrilus olivicolor Kiesenwetter was the most abundant species, while almost only Agrilus viridis (L.) was sampled by plant beating from 2008 to 2010. Moreover, almost all adults emerged from field‐collected hazel wood belonged to this latter species that proved to be the main responsible for the severe attacks on the hazel trees. Agrilus viridis adults emerged from late May to late June, and generally lived until the end of August, while egg masses were observed from late May to late July. The egg parasitoid Oobius zahaikevitshi Trjapitzin was found in all of the investigated groves; adults emerged already from the first egg masses collected on hazel trees in late May‐early June. O. zahaikevitshi was able to largely reduce A. viridis populations, with a parasitism rate of more than 50% in some groves. Recent attacks of A. viridis were related to a long period of drought; however, appropriate agronomic practices to minimize the effects of water stress and to conserve natural enemies, such as O. zahaikevitshi, could be more effective to protect hazel groves against boring beetle attacks.     

Fluorescent prey traps in carnivorous plants

Carnivorous plants acquire most of their nutrients by capturing ants, insects and other arthropods through their leaf‐evolved biological traps. So far, the best‐known attractants in carnivorous prey traps are nectar, colour and olfactory cues. Here, fresh prey traps of 14 Nepenthes, five Sarracenia, five Drosera, two Pinguicula species/hybrids, Dionaea muscipula and Utricularia stellaris were scanned at UV 366 nm. Fluorescence emissions of major isolates of fresh Nepenthes khasiana pitcher peristomes were recorded at an excitation wavelength of 366 nm. N. khasiana field pitcher peristomes were masked by its slippery zone extract, and prey capture rates were compared with control pitchers. We found the existence of distinct blue fluorescence emissions at the capture spots of Nepenthes, Sarracenia and Dionaea prey traps at UV 366 nm. These alluring blue emissions gradually developed with the growth of the prey traps and diminished towards their death. On excitation at 366 nm, N. khasiana peristome 3:1 CHCl₃–MeOH extract and its two major blue bands showed strong fluorescence emissions at 430–480 nm. Masking of blue emissions on peristomes drastically reduced prey capture in N. khasiana pitchers. We propose these molecular emissions as a critical factor attracting arthropods and other visitors to these carnivorous traps. Drosera, Pinguicula and Utricularia prey traps showed only red chlorophyll emissions at 366 nm.     

Antimicrobial resistance and toxin gene profiles of Staphylococcus aureus strains from Holstein milk

Isolation of Staphylococcus aureus (Staph. aureus) from Holstein milk samples with mastitis and nonmastitis was conducted to estimate its prevalence, antimicrobial resistance and toxin genes. A total of 353 milk samples were collected from three Chinese Holstein herds. Fifty‐three Staph. aureus isolates collected from 29 Staph. aureus‐positive samples were characterized via antimicrobial susceptibility, toxin genes and Pulsed‐field Gel Electrophoresis (PFGE) profiles. The prevalence of Staph. aureus was 4·0–9·5% in mastitic and 7·3–11·5% in nonmastitic samples in the analysed herds. Approximately 61·0% of Staph. aureus strains isolated from mastitis cows were resistant to ≥10 antimicrobials compared with 0% of isolates with nonmastitis. The most frequently observed super antigenic toxin gene was pvl (41·5%) followed by seh + pvl (13·2%). We did not find mecA‐positive methicillin‐resistant Staph. aureus (MRSA) strains, while mecA‐negative MRSA strains were identified in the three herds. PFGE results suggested potential transmission of Staph. aureus strains in different farms. These results open new insights into Staph. aureus transmission and antimicrobial resistance of Holstein dairy cows and into developing strategies for udder health improvement of dairy cattle.     

Presence of Cry1Ab in the Bt maize – aphid (Rhopalosiphum maidis) – ladybeetle (Propylea japonica) system has no adverse effects on insect biological parameters

Transgenic Bacillus thuringiensis Berliner (Bt) crops receive particular attention because they carry genes encoding insecticidal proteins that might negatively affect non‐target arthropods. Here, laboratory experiments were conducted to evaluate the impact of Cry1Ab‐expressing transgenic maize [5422Bt1 (event Bt11) and 5422CBCL (MON810)] on the biological parameters of two non‐target arthropods, the aphid Rhopalosiphum maidis (Fitch) (Hemiptera: Aphididae) and its predator the ladybeetle Propylea japonica (Thunberg) (Coleoptera: Coccinellidae). In a long‐term assay (three generations), no significant differences were found between R. maidis fed Bt maize and those fed a near‐isogenic line (5422) when individual parameters were compared, including nymph development time, adult longevity, aphid spawning period, and fecundity. No negative effects were detected throughout the life cycle of P. japonica in aphids’ feeding amount, development (nymphs, pupae, adults, and progeny eggs), fecundity, or egg hatching when they preyed on Bt maize‐fed aphids compared with non‐Bt maize treatments. A tritrophic assay revealed that Cry1Ab was highly diluted through the food chain (Bt maize leaves, R. maidis, and P. japonica), as detected by an enzyme‐linked immunosorbent assay (ELISA). In conclusion, although Cry1Ab concentrations in maize leaves increased as the plants developed, Cry1Ab levels were significantly reduced in the aphid R. maidis, and no traces of Cry1Ab were detected in P. japonica preying on Bt maize‐fed aphids. The two hybrids of Bt maize expressing Cry1Ab had no negative effects on the measured biological parameters of the aphid R. maidis or its predator, the ladybeetle P. japonica.     

Orthogonal array design for optimization of an artificial medium for in vitro rearing of Trichogramma dendrolimi

Successful complete development of Trichogramma species on artificial media is also related to the presence and proportions of ingredients other than host insect‐derived components. In this study, an orthogonal array with six factors at three levels was performed and parameters of parasitism, larval development, pupation, and adult emergence were monitored to reveal the most important components of an artificial medium and to improve the medium for in vitro mass rearing of Trichogramma dendrolimi Matsumura (Hymenoptera: Trichogrammatidae). Results indicated that biological parameters of T. dendrolimi were affected differently by six ingredients of the artificial medium: pupal hemolymph of Antheraea pernyi (Guérin‐Méneville) (Lepidoptera: Saturniidae), egg yolk, 10% malted milk solution, Neisenheimer's salt solution, trehalose, and sterile water. Statistical analysis indicated that trehalose and Neisenheimer's salt solution, 10% malted milk solution, and pupal hemolymph of A. pernyi were the main ingredients of the artificial medium based on rates of parasitism and pupation, the number of larvae developing in each artificial egg, adult emergence rate, and the number of normal adults produced. A follow‐up bioassay with a selection of optimized formulas confirmed the validity of the optimization as predicted by the orthogonal array analysis, indicating the usefulness of this method for selecting artificial diets for entomophagous insects. Adult emergence rate of the parasitoid and total number of normal adults produced per egg card (each containing 20 artificial eggs) averaged 88.8% and 956 females on the best performing optimized artificial medium, consisting of 3 ml pupal hemolymph of A. pernyi, 2.5 ml egg yolk, 1 ml 10% malted milk solution, 1 ml Neisenheimer's salt solution, 0.1 g trehalose, and 1.5 ml sterile water. The latter medium was superior to any formerly developed medium and may thus have potential for the in vitro mass rearing of T. dendrolimi.     

Floral volatiles with colour cues from two cucurbitaceous plants causing attraction of Aulacophora foveicollis

Aulacophora foveicollis Lucas (Coleoptera: Chrysomelidae) is an important phytophagous pest of two cucurbitaceous plants, Momordica cochinchinensis Spreng and Solena amplexicaulis (Lam.) Gandhi. The volatile organic compound profiles from flowers of M. cochinchinensis and S. amplexicaulis were identified and quantified by gas chromatography‐mass spectrometry (GC‐MS) and GC‐flame ionization detector (FID) analyses. Twenty nine and 28 compounds were identified in volatiles of M. cochinchinensis and S. amplexicaulis flowers, respectively. Methyl jasmonate and 3‐octanol were the predominant volatiles of M. cochinchinensis flowers, whereas 1‐octadecanol and 1‐hexanol were most found in the headspace of S. amplexicaulis flowers. Aulacophora foveicollis were more attracted by the flower volatiles of M. cochinchinensis than by those of S. amplexicaulis in a glass Y‐tube olfactometer. A mixture of 1‐heptanol, linalool oxide, 1‐octanol, and nonanal in the proportions present in the headspace of both flower types elicited attraction in the insect. From 25 cm distance, A. foveicollis displayed a preference for artificial flowers of 6.5 cm diameter of S. amplexicaulis flower colour (white) over M. cochinchinensis flower colour (white‐yellow). Finally, a synthetic blend (0.43 μg 1‐heptanol + 1.44 μg linalool oxide + 0.14 μg 1‐octanol + 1.77 μg nonanal dissolved in 25 μl methylene chloride) attracted more beetles when applied in a white artificial flower than when applied in a white‐yellow artificial flower from 40 cm distance. This finding may be helpful in the development of traps for pest management strategies.     

Variation in Polyphenolic Profiles,Antioxidant and Antimicrobial Activity of Different Achillea Species as Natural Sources of Antiglycative Compounds

A comparative study was carried out on the methanolic extracts from six Achillea species and the examined polyphenols from these plants on the formation of advanced glycation end‐products (AGE) in vitro. A. pachycephala which was richer in flavonoids (15 mg quercetin/g W) and phenolics (111.10 mg tannic acid/g DW) with substantial antioxidant activity (IC₅₀ = 365.5 μg/ml) presented strong anti‐AGE properties. Chlorogenic acid, luteolin, quercetin and caffeic acid were identified as the major polyphenols in the extracts by HPLC. In general, polyphenolic content follows the order of A. pachycephalla > A. nobilis > A. filipendulina > A. santolina > A. aucheri > A. millefolium. Most extracts exhibited marked anti‐AGE ability in the bovine serum albumin (BSA)/methylglyoxal (MG) system, though A. pachycephala showed the highest potential. The formation of AGEs was assessed by monitoring the production of fluorescent products and circular dichroism (CD) spectroscopy. Diminution in free radical production (assessed by 2,2‐diphenyl‐1‐picrylhydrazyl (DPPH) assays) is discussed as potential mechanism for delay or reduced AGE. The results demonstrate the antiglycative, antioxidant and antimicrobial potential of Achillea species which can be attributed to polyphenols content and the effectiveness on generation of AGEs, thus Achillea species can be considered as natural sources for slowing down glycation related diseases.     

Horizontal transmission of a microsporidium from the convergent lady beetle, Hippodamia convergens Guérin-Méneville (Coleoptera: Coccinellidae), to three coccinellid species of Nova Scotia

Convergent lady beetles, Hippodamia convergens Guérin-Méneville, are a popular choice for aphid control in North America. An unidentified microsporidium was found in H. convergens adults that were purchased from a commercial insectary in 2004. This study examined egg cannibalism and egg predation as a means of horizontal transmission of the unidentified microsporidium among H. convergens larvae and three coccinellid species found in Nova Scotia: Coccinella septempunctata (seven-spotted lady beetle), C. trifasciata perplexa (three-banded lady beetle), and Harmonia axyridis (multicolored Asian lady beetle). The microsporidium was transmitted with 100% efficiency when first instars fed on microsporidia-infected eggs. Mean spore count data from smear preparations of infected beetles suggest that the infection was as heavy in C. trifasciata perplexa (a native coccinellid) (11.2 ± 0.96 spores/100 μm²) as it was in H. convergens (the natural host) (12.8 ± 1.16) but lighter in the introduced species C. septempunctata (7.5 ± 0.65) and H. axyridis (0.8 ± 0.11). For all of the beetle species examined, larval development was significantly longer for microsporidia-infected individuals than for their uninfected cohorts. The microsporidium had no effect on larval mortality. Based on the results of this study, field-collected H. convergens should be examined for microsporidia and uninfected individuals should be used to rear individuals for release in biological control programs. However, this is unlikely to happen because H. convergens are relatively easy and inexpensive to collect from their overwintering sites for redistribution.     

New Middle Cambrian bivalved arthropods from the Burgess Shale (British Columbia,Canada)

The morphology of two new bivalved arthropods, Loricicaris spinocaudatus gen. et sp. nov. and Nereocaris briggsi sp. nov. from the middle Cambrian (Series 3, Stage 5) Burgess Shale Formation (Collins Quarry locality on Mount Stephen, Yoho National Park, British Columbia, Canada), is described. The material was originally assigned to the genus Branchiocaris, but exhibits distinctive character combinations meriting its assignment to other taxa. Loricicaris spinocaudatus possesses an elongate and spinose abdomen comparable to the contemporaneous Perspicaris and Canadaspis, as well as chelate second head appendages and subtriangular exopods, comparable to Branchiocaris. Nereocaris briggsi possesses a laterally compressed carapace, elongate and delicate appendages and a medial eye located between a pair of lateral eyes on a rhomboidal eye stalk. Although undoubtedly congeneric with Nereocaris exilis from a slightly younger horizon of the Burgess Shale Formation, N. briggsi differs in overall proportions and segment number, warranting assignment to a new species. The newly described taxa were coded into an extensive cladistic analysis of 755 characters, and 312 extinct and extant panarthropods, including a variety of Cambrian bivalved arthropods from both the Burgess Shale and the Chengjiang Lagerstätten. Cambrian bivalved arthropods consistently resolved as a paraphyletic assemblage at the base of Arthropoda. Important innovations in arthropod history such as the specialization of the deutocerebral head appendages and a shift from a nekton‐benthic deposit feeding habit to a benthic scavenging/predatory habit, the symplesiomorphic feeding condition of Euarthropoda (crown‐group arthropods), were found to have occurred among basal bivalved arthropods.