1000 spider silkomes: Linking sequences to silk physical properties.

Spider silks are among the toughest known materials and thus provide models for renewable, biodegradable, and sustainable biopolymers. However, the entirety of their diversity still remains elusive, and silks that exceed the performance limits of industrial fibers are constantly being found. We obtained transcriptome assemblies from 1098 species of spiders to comprehensively catalog silk gene sequences and measured the mechanical, thermal, structural, and hydration properties of the dragline silks of 446 species. The combination of these silk protein genotype-phenotype data revealed essential contributions of multicomponent structures with major ampullate spidroin 1 to 3 paralogs in high-performance dragline silks and numerous amino acid motifs contributing to each of the measured properties. We hope that our global sampling, comprehensive testing, integrated analysis, and open data will provide a solid starting point for future biomaterial designs.

Phylogenomic Analysis of Ultraconserved Elements Resolves the Evolutionary and Biogeographic History of Segmented Trapdoor Spiders.

The segmented trapdoor spiders (Liphistiidae) are the sole surviving family of the suborder Mesothelae, which forms the sister lineage to all other living spiders. Liphistiids have retained a number of plesiomorphic traits and their present-day distribution is limited to East and Southeast Asia. Studying this group has the potential to shed light on the deep evolutionary history of spiders, but the phylogeny and divergence times of the family have not been resolved with confidence. We performed phylogenomic and molecular dating analyses of 2765 ultraconserved element loci from 185 liphistiid taxa. Our analyses show that the crown group of Liphistiidae appeared in the mid-Cretaceous at 102 Ma (95% credibility interval 92-113 Ma), but it was not until the Neogene that much of the diversification within the family occurred in mainland Southeast and East Asia. This diversification was coincident with tectonic events such as the extension of the East Asian continental margin, as well as geological upheavals in Indochina induced by the collision between India and Asia. Our study highlights the important role of major tectonic events in shaping the evolutionary history, present-day diversity, and geographical distribution of mesothele and liphistiid spiders. [biogeography; concatenation; Liphistiidae; molecular dating; summary coalescent; UCEs.].

Female spider aggression is associated with genetic underpinnings of the nervous system and immune response to pathogens.

Identifying the genetic architecture underlying phenotypic variation in natural populations and assessing the consequences of polymorphisms for individual fitness are fundamental goals in evolutionary and molecular ecology. Consistent between-individual differences in behaviour have been documented for a variety of taxa. Dissecting the genetic basis of such behavioural differences is however a challenging endeavour. The molecular underpinnings of natural variation in aggression remain elusive. Here, we used comparative gene expression (transcriptome analysis and RT-PCR), genetic association analysis and pharmacological experiments to gain insight into the genetic basis of aggression in wild-caught jumping spiders (Portia labiata). We show that spider aggression is associated with a putative viral infection response gene, BTB/POZ domain-containing protein 17 (BTBDH), in addition to a putative serotonin receptor 1A (5-HT1A) gene. Spider aggression varies with virus loads, and BTBDH is upregulated in docile spiders and exhibits a genetic variant associated with aggression. We also identify a putative serotonin receptor 5-HT1A gene upregulated in docile P. labiata. Individuals that have been treated with serotonin become less aggressive, but individuals treated with a nonselective serotonin receptor antagonist (methiothepin) also reduce aggression. Further, we identify the genetic variants in the 5-HT1A gene that are associated with individual variation in aggression. We therefore conclude that co-evolution of the immune and nervous systems may have shaped the between-individual variation in aggression in natural populations of jumping spiders.

Organophosphate and Organochlorine Resistance in Larval Stage of Aedes albopictus (Diptera: Culicidae) in Sabah, Malaysia.

The present study aims to investigate the susceptibility status of Aedes albopictus (Skuse) collected from residential areas in Sabah, Malaysia towards eight WHO-recommended dosages of larvicides representing the classes of organophosphates and organochlorines. Field and reference strains of Ae. albopictus larvae were bioassayed in accordance to WHO standard methods using diagnostic dosages of bromophos, malathion, fenthion, fenitrothion, temephos, chlorpyrifos, dichlorodiphenyltrichloroethane (DDT), and dieldrin. The results revealed that Ae. albopictus was resistant (mortality < 90%) towards malathion, temephos, and DDT. In addition, most of the Ae. albopictus strains exhibited a wide range of susceptibilities against bromophos, with mortality ranged from 49.33 to 93.33%. On the contrary, only dieldrin was able to induce 100% mortality against all strains of Ae. albopictus. Tolerance to fenitrothion, fenthion, and chlorpyrifos, with mortality ranging from 81.33 to 97.33%, was also observed in this study.

Morphological and mechanical properties of flexible resilin joints on damselfly wings (Rhinocypha spp.).

Resilin functions as an elastic spring that demonstrates extraordinary extensibility and elasticity. Here we use combined techniques, laser scanning confocal microscopy (LSCM) and scanning electron microscopy (SEM) to illuminate the structure and study the function of wing flexibility in damselflies, focusing on the genus Rhinocypha. Morphological studies using LSCM and SEM revealed that resilin patches and cuticular spikes were widespread along the longitudinal veins on both dorsal and ventral wing surfaces. Nanoindentation was performed by using atomic force microscopy (AFM), where the wing samples were divided into three sections (membrane of the wing, mobile and immobile joints). The resulting topographic images revealed the presence of various sizes of nanostructures for all sample sections. The elasticity range values were: membrane (0.04 to 0.16 GPa), mobile joint (1.1 to 2.0 GPa) and immobile joint (1.8 to 6.0 GPa). The elastomeric and glycine-rich biopolymer, resilin was shown to be an important protein responsible for the elasticity and wing flexibility.

Predator personality and prey behavioural predictability jointly determine foraging performance.

Predator-prey interactions play important roles in ecological communities. Personality, consistent inter-individual differences in behaviour, of predators, prey or both are known to influence inter-specific interactions. An individual may also behave differently under the same situation and the level of such variability may differ between individuals. Such intra-individual variability (IIV) or predictability may be a trait on which selection can also act. A few studies have revealed the joint effect of personality types of both predators and prey on predator foraging performance. However, how personality type and IIV of both predators and prey jointly influence predator foraging performance remains untested empirically. Here, we addressed this using a specialized spider-eating jumping spider, Portia labiata (Salticidae), as the predator, and a jumping spider, Cosmophasis umbratica, as the prey. We examined personality types and IIVs of both P. labiata and C. umbratica and used their inter- and intra-individual behavioural variation as predictors of foraging performance (i.e., number of attempts to capture prey). Personality type and predictability had a joint effect on predator foraging performance. Aggressive predators performed better in capturing unpredictable (high IIV) prey than predictable (low IIV) prey, while docile predators demonstrated better performance when encountering predictable prey. This study highlights the importance of the joint effect of both predator and prey personality types and IIVs on predator-prey interactions.

Biting behaviour of medically important mosquitoes (Diptera: Culicidae) in Peninsular Malaysia.

Human-landing catch (HLC) technique was undertaken in nine selected study sites within Peninsular Malaysia to determine the biting cycle of Aedes, Armigeres, Culex, and Mansonia mosquitoes. HLC was conducted 24 hours on three different nights in a duration of one week at each study site. Aedes albopictus were found in all study sites with bimodal biting cycle, whereas the biting peak of Aedes butleri was recorded between 1600 and 1900 hr. For Armigeres, five study sites demonstrated similar biting peak hour (1900 - 2000 hr) for Ar. kesseli four study sites showed biting peak at 1800 - 1900 hr for Ar. subalbatus. Culex quinquefasciatus was collected from all study sites except in Sungai Penchala with its biting activity beginning from 1900 hr. Both Mansonia annulifera and Mansonia uniformis captured in two different study sites showed similar biting peaks (1500 - 1600 hr). It is recommended that vector control activities be carried out at dusk as almost all mosquito species captured started their biting activities at that time.

Biting behaviour of medically important mosquitoes (Diptera: Culicidae) in Peninsular Malaysia

Human-landing catch (HLC) technique was undertaken in nine selected study sites within Peninsular Malaysia to determine the biting cycle of Aedes, Armigeres, Culex, and Mansonia mosquitoes. HLC was conducted 24 hours on three different nights in a duration of one week at each study site. Aedes albopictus were found in all study sites with bimodal biting cycle, whereas the biting peak of Aedes butleri was recorded between 1600 and 1900 hr. For Armigeres, five study sites demonstrated similar biting peak hour (1900 – 2000 hr) for Ar. kesseli four study sites showed biting peak at 1800 – 1900 hr for Ar. subalbatus. Culex quinquefasciatus was collected from all study sites except in Sungai Penchala with its biting activity beginning from 1900 hr. Both Mansonia annulifera and Mansonia uniformis captured in two different study sites showed similar biting peaks (1500 – 1600 hr). It is recommended that vector control activities be carried out at dusk as almost all mosquito species captured started their biting activities at that time.

A multi-locus approach resolves the phylogenetic relationships of the Simulium asakoae and Simulium ceylonicum species groups in Malaysia: evidence for distinct evolutionary lineages.

A multi-locus approach was used to examine the DNA sequences of 10 nominal species of blackfly in the Simulium subgenus Gomphostilbia (Diptera: Simuliidae) in Malaysia. Molecular data were acquired from partial DNA sequences of the mitochondria-encoded cytochrome c oxidase subunit I (COI), 12S rRNA and 16S rRNA genes, and the nuclear-encoded 18S rRNA and 28S rRNA genes. No single gene, nor the concatenated gene set, resolved all species or all relationships. However, all morphologically established species were supported by at least one gene. The multi-locus sequence analysis revealed two distinct evolutionary lineages, conforming to the morphotaxonomically recognized Simulium asakoae and Simulium ceylonicum species groups.

Extant primitively segmented spiders have recently diversified from an ancient lineage.

Living fossils are lineages that have retained plesiomorphic traits through long time periods. It is expected that such lineages have both originated and diversified long ago. Such expectations have recently been challenged in some textbook examples of living fossils, notably in extant cycads and coelacanths. Using a phylogenetic approach, we tested the patterns of the origin and diversification of liphistiid spiders, a clade of spiders considered to be living fossils due to their retention of arachnid plesiomorphies and their exclusive grouping in Mesothelae, an ancient clade sister to all modern spiders. Facilitated by original sampling throughout their Asian range, we here provide the phylogenetic framework necessary for reconstructing liphistiid biogeographic history. All phylogenetic analyses support the monophyly of Liphistiidae and of eight genera. As the fossil evidence supports a Carboniferous Euramerican origin of Mesothelae, our dating analyses postulate a long eastward over-land dispersal towards the Asian origin of Liphistiidae during the Palaeogene (39-58 Ma). Contrary to expectations, diversification within extant liphistiid genera is relatively recent, in the Neogene and Late Palaeogene (4-24 Ma). While no over-water dispersal events are needed to explain their evolutionary history, the history of liphistiid spiders has the potential to play prominently in vicariant biogeographic studies.

TEMPORAL CHANGES OF AEDES AND ARMIGERES POPULATIONS IN SUBURBAN AND FORESTED AREAS IN MALAYSIA.

Surveillance of mosquitoes and their distribution in association with rainfall, relative humidity, and temperature were conducted in selected suburban and forested areas, namely, Sungai Penchala (Kuala Lumpur) and Taman Alam (Selangor) for 12 months. Armigeres kesseli was the most abundant species in Sungai Penchala while Aedes butleri was the most dominant species in Taman Alam. A positive correlation between mosquito distribution and rainfall was observed in selected mosquito species in Sungai Penchala (Armigeres kesseli, r = 0.75; Armigeres subalbatus, r = 0.62; and Aedes albopictus, r = 0.65) and Taman Alam (Armigeres sp, r = 0.59; Ae. butleri, r = 0.85; and Ae. albopictus, r = 0.62). However, no significant cor- relation was found either between selected mosquito species in both study areas and relative humidity or temperature. Results obtained suggested that vector control programs to be conducted based on temporal distribution of vectors in order to achieve beneficial outcomes with effective costing.

Guppies as predators of common mosquito larvae in Malaysia.

Observation on predation activities of guppies (Poecilia reticulata) on the larvae of three species of mosquito, namely Aedes albopictus, Aedes aegypti, and Culex quinquefasciatus was carried out under laboratory conditions. Male and female guppies were used as predators for predation experiments on the 4th instars of mosquito larvae. The daily feeding rates comparing male and female guppies on mosquito larvae were different; the female guppies consumed more mosquito larvae than male guppies did. The daily feeding rates of female guppies were 121.3 for Ae. aegypti, 105.6 for Ae. albopictus, and 72.3 for Cx. quinquefasciatus. The daily feeding rates of male guppies were 98.6 for Ae. aegypti, 73.6 for Ae. albopictus, and 47.6 for Cx. quinquefasciatus. In terms of prey preference, there was greater preference towards mosquito larvae of Ae. aegypti, followed by Ae. albopictus, and the least preferred was Cx. quinquefasciatus. Male and female guppies consumed more mosquito larvae during lights on (day time) compared with lights off (night time). The water volume, prey species, number of fish predators available, prey densities, and prey's sex also influenced the predation activities.

Mitochondrial DNA analyses reveal low genetic diversity in Culex quinquefasciatus from residential areas in Malaysia.

The present study explored the intraspecific genetic diversity, dispersal patterns and phylogeographic relationships of Culex quinquefasciatus Say (Diptera: Culicidae) in Malaysia using reference data available in GenBank in order to reveal this species' phylogenetic relationships. A statistical parsimony network of 70 taxa aligned as 624 characters of the cytochrome c oxidase subunit I (COI) gene and 685 characters of the cytochrome c oxidase subunit II (COII) gene revealed three haplotypes (A1-A3) and four haplotypes (B1-B4), respectively. The concatenated sequences of both COI and COII genes with a total of 1309 characters revealed seven haplotypes (AB1-AB7). Analysis using tcs indicated that haplotype AB1 was the common ancestor and the most widespread haplotype in Malaysia. The genetic distance based on concatenated sequences of both COI and COII genes ranged from 0.00076 to 0.00229. Sequence alignment of Cx. quinquefasciatus from Malaysia and other countries revealed four haplotypes (AA1-AA4) by the COI gene and nine haplotypes (BB1-BB9) by the COII gene. Phylogenetic analyses demonstrated that Malaysian Cx. quinquefasciatus share the same genetic lineage as East African and Asian Cx. quinquefasciatus. This study has inferred the genetic lineages, dispersal patterns and hypothetical ancestral genotypes of Cx. quinquefasciatus.

Temephos resistance in field Aedes (Stegomyia) albopictus (Skuse) from Selangor, Malaysia.

Larvae of Aedes albopictus obtained from dengue endemic areas in Selangor, Malaysia were evaluated for their susceptibility to operational dosage of temephos (1 mg/L). Larval bioassays were carried out in accordance to modified WHO standard methods. Biochemical microassay of enzymes in Ae. albopictus was conducted to detect the emergence of insecticide resistance and to define the mechanisms involved in temephos resistance. The 50% mortality lethal time (LT50) for Ae. albopictus tested against temephos ranged between 58.65 to 112.50 minutes, with resistance ratio ranging from 0.75 - 1.45. This study addressed the fluctuation of time-related susceptibility status of Ae. albopictus towards insecticide. Significant difference on the weekly enzyme levels of non-specific esterases, mixed function oxidases and glutathione S-transferases was detected (p ≤ 0.05). No significant correlation was found between temephos resistance and enzyme activity (p > 0.05). Only glutathione S-transferases displayed high level of activity, indicating that Ae. albopictus may be resistant to other groups of insecticide. The insensitive acetylcholinesterase was detected in some field collected Ae. albopictus populations, indicating the possibility of emergence of carbamate or other organophosphate resistance in the field populations. Continuous resistance monitoring should be conducted regularly to confirm the efficacy of insecticides for dengue control.

Mosquito species and outdoor breeding places in residential areas in Malaysia.

We conducted mosquito surveillance at outdoor breeding habitat in 459 households at 7 urban locations in Putrajaya, Malaysia from January to December 2010 to determine the predominant species and breeding locations. The most common species found at all locations was Aedes albopictus. Gardening utensils were the most common breeding sites. Of the 1,885 mosquito larvae specimens found, 1,774 (94.1%) were Ae. albopictus larvae, 84 (4%) were Ae. aegypti larvae and 27 (1%) were Culex quinquefasciatus larvae. The Aedes index for each of the locations was higher than the goal set by the Ministry of Health for Malaysia. However, the container index at each of the locations was within the goal. The Breateau index was above the goal set by the Ministry of Health at Precinct 9B1 but the other locations were within the goal.