Trypanosomatids are obligate parasites of animals, predominantly insects and vertebrates, and flowering plants. Monoxenous species, representing the vast majority of trypanosomatid diversity, develop in a single host, whereas dixenous species cycle between two hosts, of which primarily insect serves as a vector. To explore in-depth the diversity of insect trypanosomatids including their co-infections, sequence profiling of their 18S rRNA gene was used for true bugs (Hemiptera; 18% infection rate) and flies (Diptera; 10%) in Cuba. Out of 48 species (molecular operational taxonomic units) belonging to the genera Vickermania (16 spp.), Blastocrithidia (7), Obscuromonas (4), Phytomonas (5), Leptomonas/Crithidia (5), Herpetomonas (5), Wallacemonas (2), Kentomonas (1), Angomonas (1) and two unnamed genera (1 + 1), 38 species have been encountered for the first time. The detected Wallacemonas and Angomonas species constitute the most basal lineages of their respective genera, while Vickermania emerged as the most diverse group. The finding of Leptomonas seymouri, which is known to rarely infect humans, confirms that Dysdercus bugs are its natural hosts. A clear association of Phytomonas with the heteropteran family Pentatomidae hints at its narrow host association with the insect rather than plant hosts. With a focus on multiple infections of a single fly host, using deep Nanopore sequencing of 18S rRNA, we have identified co-infections with up to 8 trypanosomatid species. The fly midgut was usually occupied by several Vickermania species, while Herpetomonas and/or Kentomonas species prevailed in the hindgut. Metabarcoding was instrumental for analysing extensive co-infections and also allowed the identification of trypanosomatid lineages and genera.

Methoprene, a juvenile hormone analog, is used to accelerate sexual maturation in males of species of economic importance in support to the sterile insect technique (SIT). In the SIT, mass-reared sterile males are released into the field and need to survive until they reach sexual maturation, find a wild female, mate with her and then induce female sexual refractoriness, so she will not remate with a wild counterpart. The use of methoprene shortens the time between release and copulation. However, in South American fruit flies, Anastrepha fraterculus, the ability of methoprene-treated males to inhibit female remating has been shown to be lower than wild males, when methoprene was applied by pupal immersion or topical application. Here we evaluated the possibility of incorporating methoprene into the male diet at different doses and the ability of those males to inhibit female remating, as well as the effect of methoprene on male reproductive organ size, due to the possible correlation between male accessory gland size and their content, and the role of male accessory gland proteins in female inhibition. We found that A. fraterculus males fed with methoprene in the adult protein diet at doses as high as 1% were less likely to inhibit female remating, however, at all other lower doses males had the same ability as untreated males to inhibit female remating. Males fed with methoprene had bigger male accessory glands and testes compared to methoprene-deprived males. We demonstrate that the incorporation of methoprene in adult male diets is possible in this species and potentially useful as a post-teneral, pre-release supplement at doses as low as 0.01%. Even at higher doses, the percentage of females remating after 48 h from the first copulation is sufficiently low in this species so as not compromise the efficiency of the SIT.

Interaction networks can provide detailed information regarding ecological systems, helping us understand how communities are organized and species are connected. The goals of this study were to identify the pattern of interaction between bats and ectoparasites in urban green areas of Grande Aracaju, Sergipe, and calculate connectance, specialization, nesting, modularity and centrality metrics. Bats were captured using 10 mist nets inside and on the edges of the fragments, and the collected ectoparasites were stored in 70% alcohol. All analyses were performed using R software. The interaction network consisted of 10 species of bats and 13 ectoparasites. Connectivity was considered low (0.12). The specialization indices for ectoparasites ranged from 0.50 to 1.00, and the value obtained for the network was 0.96, which is high. The observed nesting metric was low (wNODF = 1.47), whereas the modularity was high (wQ = 0.74), indicating that the studied network had a modular topology. All centrality metrics had low values. The observed modularity may have been caused by the evolutionary history of the bats and ectoparasites involved and the high specificity index of the interactions. The low centrality values may be associated with low connectivity and a high degree of specialization. This study provides relevant information on bat–parasite interactions in an urban environment, highlighting the need for further studies to improve our understanding of host–parasite interaction networks.

Sampling flower-visiting insects in agricultural fields at large spatial and temporal scales is significant for understanding local insect pollinator communities. The most commonly used method, pan trap, has been criticized due to its attractant bias. A window trap (also referred to as the flight-intercept trap) is a non-attractant sampling method, which has been applied in forests and grasslands, but rarely in agricultural fields. We aim to test whether we can replace pan traps with window traps in agricultural fields by comparing species richness and species composition between the two methods, and to show whether flower-visiting insects collected in both traps can reflect flower-visiting activity recorded by camera observation. We conducted a 2-year study to compare the performance of these sampling methods in an oilseed rape field. Results showed that the relative abundance of dominant flower-visiting species was highly correlated between the window trap and the pan trap samples, while window traps caught more individuals and higher (rarefied) species richness than pan traps. The species composition of window traps was more similar to each other than that of pan traps. The proportion of honey bees (Apis spp.) collected in both traps underestimated their flower-visiting activity recorded by camera observations, while sweat bees (Halictidae) and butterflies (Lepidoptera) were overestimated. Our study suggests that the window trap has the potential to serve as an alternative sampling method of flower-visiting insects to the pan trap. However, we need to be cautious when using specimens caught in both traps as a proxy of their flower-visiting activity.

Many parasitoid species discriminate already parasitized hosts, thus avoiding larval competition. However, females incur in superparasitism under certain circumstances. Superparasitism is commonly observed in the artificial rearing of the parasitoid Diachasmimorpha longicaudata, yet host discrimination has been previously suggested in this species. Here, we addressed host discrimination in virgin D. longicaudata females in a comprehensive way by means of direct and indirect methods, using Ceratitis capitata and Anastrepha fraterculus which are major fruit fly pests in South America. Direct methods relied on the description of the foraging behaviour of females in arenas with parasitized and non-parasitized host larvae. In the indirect methods, healthy larvae were offered to single females and the egg distributions were compared to a random distribution. We found that D. longicaudata was able to recognize parasitized host from both host species, taking 24 h since a first parasitization for A. fraterculus and 48 h for C. capitata. Indirect methods showed females with different behaviours for both host species: complete discrimination, non-random (with superparasitism), and random distributions. A larger percentage of females reared and tested on A. fraterculus incurred in superparasitism, probably associated with higher fecundity. In sum, we found strong evidence of host discrimination in D. longicaudata, detecting behavioural variability associated with the host species, the time since the first parasitization and the fecundity of the females.

A survey was conducted at 88 sites in Montana in 1984 and 1985 to determine the distribution of Urophora affinis Frauenfeld and U. quadrifasciata (Meigen), two seed head flies released on spotted knapweed (Centaurea maculosa Lam. # CENMA) in North America. U. affinis, released in Montana during 1973 to 1977, was found at 40 of the 88 sites. The fly was concentrated within a short radius (about 5 to 8 km) of release sites, even at sites having well-established populations. U. quadrifasciata, released in British Columbia in 1972, but not in the United States, was found at 84 sites. The data suggest that the fly entered the state in the northwest corner and dispersed in a southeasterly direction. There was a strong positive relationship between mean number of galls per seed head and percent infestation for both fly species.

Campylobacter is the leading cause of bacterial diarrhoeal disease worldwide, with raw and undercooked poultry meat and products the primary source of infection. Colonization of broiler chicken flocks with Campylobacter has proved difficult to prevent, even with high levels of biosecurity. Dipteran flies are proven carriers of Campylobacter and their ingress into broiler houses may contribute to its transmission to broiler chickens. However, this has not been investigated in the UK. Campylobacter was cultured from 2195 flies collected from four UK broiler farms. Of flies cultured individually, 0·22% [2/902, 95% confidence interval (CI) 0–0·53] were positive by culture for Campylobacter spp. Additionally, 1293 flies were grouped by family and cultured in 127 batches: 4/127 (3·15%, 95% CI 0·11-6·19) from three broiler farms were positive for Campylobacter. Multilocus sequence typing of isolates demonstrated that the flies were carrying broiler-associated sequence types, responsible for human enteric illness. Malaise traps were used to survey the dipteran species diversity on study farms and also revealed up to 612 flies present around broiler-house ventilation inlets over a 2-h period. Therefore, despite the low prevalence of Campylobacter cultured from flies, the risk of transmission by this route may be high, particularly during summer when fly populations are greatest.

Parasitoids are significant enemies of many economically important insects and there is some evidence to suggest that their actions have a role in terminating the outbreaks of forest Lepidoptera populations. In this study, we examined the impact of parasitoids on the pupae of the pine processionary moth, and highlighted the presence of several parasitoid species for this developmental stage. A higher rate of parasitism was found when the pupal density in the soil was reduced, but the rate of parasitism was not influenced by pupal morphological traits or by the presence or absence of a cocoon around a pupa. Of the external factors examined, a delay in the time of descent of larvae from the trees had a positive effect on the level of parasitism. Observational data indicated that dipteran and hymenopteran were the most abundant parasitoids to emerge from moth pupae. Our study highlights the complexity of the parasitoid–host dynamics, and stresses the importance of carefully determining environmental effects on host–parasitoid relations.

The adaptive significance of variation in sperm phenotype is still largely unknown, in part due to the difficulties of observing and measuring sperm movement in its natural, selective environment (i.e., within the female reproductive tract). Computer-assisted sperm analysis systems allow objective and accurate measurement of sperm velocity, but rely on being able to track individual sperm, and are therefore unable to measure sperm movement in species where sperm move in trains or bundles. Here we describe a newly developed computational method for measuring sperm movement using Fourier analysis to estimate sperm tail beat frequency. High-speed time-lapse videos of sperm movement within the female tract of the neriid fly Telostylinus angusticollis were recorded, and a map of beat frequencies generated by converting the periodic signal of an intensity versus time trace at each pixel to the frequency domain using the Fourier transform. We were able to detect small decreases in sperm tail beat frequency over time, indicating the method is sensitive enough to identify consistent differences in sperm movement. Fourier analysis can be applied to a wide range of species and contexts, and should therefore facilitate novel exploration of the causes and consequences of variation in sperm movement.

In this study, laboratory and field cage experiments were conducted to assess the performance of sterile mass-reared Mexican fruit fly Anastrepha ludens (Loew) irradiated at various doses. In small laboratory cages, the radiation dose was found to have no effect on adult emergence, flying adults or survival at 25 days. In field cages, non-irradiated males and males irradiated at 20 and 40 Gy were found to exhibit greater mating propensity than males irradiated at 60 and 80 Gy. In large field cages in competition with wild males, cohorts of flies (male and female) irradiated at the low dose of 40 Gy and wild non-irradiated flies mated randomly, while flies irradiated at the high standard dose of 80 Gy mated assortatively. Irrespective of the radiation dose, laboratory flies courted and attempted copulation significantly earlier in the day than wild flies. In large field cages, cohorts of flies irradiated at low doses, e.g. 40 Gy, tended to induce greater sterility into a cohort of wild flies than those irradiated at 80 Gy, but the difference was not significant. Our results suggest that lowering the radiation dose currently applied to mass-reared flies in the Mexican fruit fly eradication campaign would substantially improve male mating performance. This could result in greater sterility induction without posing a risk in areas where target pest populations naturally prevail and are under suppression during the early stages of eradication programmes.

Polar terrestrial invertebrates are suggested as being vulnerable to temperature change relative to lower latitude species, and hence possibly also to climate warming. Previous studies have shown Antarctic and Arctic Collembola and Acari to possess good heat tolerance and survive temperature exposures above 30 °C. To test this feature further, the heat tolerance and physiological plasticity of heat stress were explored in the Arctic collembolan, Megaphorura arctica, from Svalbard and the Antarctic midge, Eretmoptera murphyi, from Signy Island. The data obtained demonstrate considerable heat tolerance in both species, with upper lethal temperatures ≥35 °C (1 h exposures), and tolerance of exposure to 10 and 15 °C exceeding 56 days. This tolerance is far beyond that required in their current environment. Average microhabitat temperatures in August 2011 ranged between 5.1 and 8.1 °C, and rarely rose above 10 °C, in Ny-Ålesund, Svalbard. Summer soil microhabitat temperatures on Signy Island have previously been shown to range between 0 and 10 °C. There was also evidence to suggest that E. murphyi can recover from high-temperature exposure and that M. arctica is capable of rapid heat hardening. M. arctica and E. murphyi therefore have the physiological capacity to tolerate current environmental conditions, as well as future warming. If the features they express are characteristically more general, such polar terrestrial invertebrates will likely fare well under climate warming scenarios.

The sterile insect technique (SIT) is a non-chemical approach used to control major pests from several insect families, including Tephritidae, and entails the mass-release of sterile insects that reduce fertility of wild populations. For SIT to succeed, released sterile males must mature and compete with wild males to mate with wild females. To reach sexual maturity, the Queensland fruit fly, Bactrocera tryoni (Froggatt) (Diptera: Tephritidae), must obtain adequate nutrition after adult emergence; however, in current SIT programs sterile B. tryoni receive a pre-release diet that lacks key nutrients required to sustain sexual development. The chief objective of this study was to determine whether pre-release yeast hydrolysate (YH) supplements affect the persistence and abundance of sexually mature sterile male B. tryoni under field conditions. Experiments were run in outdoor cages under conditions of low and high environmental stress that differed markedly in temperature and humidity, and in the field. Under low environmental stress conditions, survival of sterile B. tryoni was monitored in cages under three diet treatments: (i) sugar only, (ii) sugar plus YH or (iii) sugar plus YH for 48 h and sugar only thereafter. Under high environmental stress conditions survival of sterile B. tryoni was monitored in cages under four diet treatments: (i) white sugar only, (ii) brown sugar only, (iii) white sugar plus YH and (iv) brown sugar plus YH. In a replicated field study, we released colour-marked sterile B. tryoni from two diet regimes, YH-supplemented or YH-deprived, and monitored abundance of sexually mature males. In the low-stress cage study, there was no effect of diet, although overall females lived longer than males. In the high stress cage study, mortality was lower for YH-fed flies than YH-deprived flies and females lived longer than males. In the field, YH supplementation resulted in higher abundance of sexually mature sterile males, with 1.2 YH-fed flies trapped for every YH-deprived fly trapped. Under field conditions, YH supplementation can increase over-flooding ratios and hence may improve the effectiveness of SIT programmes.

For the first time, a faunistic list of the Chironomidae fauna of Greece is presented along with its ecological distribution. Chironomidae larvae and pupal exuviae were collected from 61 locations in mainland Greece during 2006–2008. 76 taxa (genera to species) were recorded. The genera Potthastia, Brillia, Euorthocladius, Synorthocladius, Tvetenia and Xenochironomus are recorded for the first time from Greece. Furthermore, 12 species are new for Greece and five are new for mainland Greece. Chironomidae collected from running waters were analysed with ordination techniques. The results revealed that Chironomidae assemblages were distinguished into three ecoregions with distinct climatic, geological and hydrochemical features. Running waters in Northwestern Greece were generally dominated by species tolerant to water acidification. Chironomids typical of eastern uplands preferred relatively slow current and soft bottom sediments. Assemblages of the southern Peloponnese Peninsula were represented by species of relatively fast flowing, warm streams. However, these are only preliminary results and many more data are needed to justify this ecological distinction of Chironomidae assemblages. This is the first study contributing to the knowledge of a rather neglected insect group of the Greek fauna. It is expected that more species remain to be identified and recorded and future works will contribute to this initial faunal list.

Podonomus Philippi was recently found in Brazilian inselbergs. In this study, we used sequences from the barcode region of cytochrome c oxidase I (COI) to assess Podonomus molecular diversity from mountains in Southeastern Brazil. Our results revealed the adult female and the larvae of Podonomus pepinellii Roque and Trivinho-Strixino, 2004 and extended the known geographical distribution of the species. Moreover, we found high molecular divergence in two populations located in the Serra da Mantiqueira and Serra do Espinhaço mountain range when compared with the only species recorded to Brazil, indicating the possibility of two more species to be described.

The seasonality and behaviour of tabanids were studied in the Planalto Serrano of Santa Catarina State, Brazil. From March 2007 to February 2009, a total of 915 tabanid flies from 15 species and six genera were collected on an individual horse. The number of tabanid species varied from zero during autumn/winter to ten in summer (December–February). Dichelacera alcicornis Wied. (62.1%) was the most abundant species followed by Chrysops fusciapex Lutz (6.7%) and C. patricia Pechuman (5.9%). Peak catches occurred during the summer (December–March). The landing behaviour varied by genera: Chrysops (100%) preferred the head, Dichelacera (56.3%) the flanks, and Poecilodera (100%), Acanthocera (100%), Tabanus (58.8%) and Fidena (48.7%) the limbs region. This is the first work on tabanids in the Santa Catarina State, Brazil.

We investigated the pollinator assemblage in Myristica dactyloides, a dioecious tree species occupying the intermediate canopy stratum of the mid- and high-elevation wet evergreen forests and endemic to Sri Lanka and the Western Ghats of India. We surveyed two populations and, in one of them, marked four male and three female trees for observations on floral display and insect foraging, and two female trees for experimental pollination. Yellow sticky traps were used to sample insects in the canopy during the flowering season of December 2007 in addition to direct observations of insect activity in 2006. Myristicaceae members from other tropical areas have been reported to be specialized to beetle pollination, but our observations provide evidence of a generalist pollination system in M. dactyloides, composed of small, diverse insects: thysanopterans (thrips), coleopterans (beetles), halictid bees and dipterans (syrphid and phorid flies). Quantifying floral display, we found that female inflorescences were smaller, offered no reward and attracted significantly fewer pollinators in comparison to male inflorescences. Fruit set was low and could be attributed to a 29% flower abscission and abortion of young fruits, but a reasonably high natural pollination efficiency combined with pollination experiments established that there was no pollination limitation in the study population.

Rainfall seasonality can strongly influence biotic interactions by affecting host plant quality, and thus potentially regulating herbivore exposure to natural enemies. Plant defences are predicted to increase from dry to wet forests, rendering wet-forest caterpillars more vulnerable to parasitoids due to the slow-growth-high-mortality hypothesis. We collected and reared caterpillars from the understorey and trail edges of a wet forest and a seasonally dry forest to determine whether wet-forest caterpillars suffered a higher prevalence of parasitism and were less abundant than dry-forest caterpillars. In the two forests, caterpillar abundances (on average 8 h−1) and prevalence of parasitism (18%) were very similar regardless of feeding niche for both parasitism (27% versus 29% in shelter builders, and 16% versus 11% in external feeders) and caterpillar abundances (shelter builders: 1.42 versus 2.39, and external feeders: 8.27 versus 5.49 caterpillars h−1) in the dry and wet forests, respectively. A similar comparative analysis conducted in the canopy and understorey of the dry forest revealed a higher prevalence of parasitism in the canopy (43%) despite caterpillar densities similar to those in the understorey. Overall, shelter builders suffered higher parasitism than external feeders (32% versus 14.9%), and were attacked primarily by flies, whereas external feeders were more vulnerable to attack by parasitoid wasps.

Artificial neural networks (ANN) methodology, molecular analyses and comparative morphology of the male postabdomen were used successfully in parallel for species identification and resolution of some taxonomic problems concerning West Palaearctic species of the genus Tachina Meigen, 1803. Supervised feed-forward ANN with back-propagation of errors was applied on morphometric and qualitative characters to solve known taxonomic discrepancies. Background molecular analyses based on mitochondrial markers CO I, Cyt b, 12S and 16S rDNA and study of male postabdominal structures were published separately. All three approaches resolved taxonomic doubts with identical results in the following five cases: case 1, the four presently recognized subgenera of the genus Tachina were confirmed and the description of a new subgenus was recommended; case 2, the validity of a new boreo-alpine species (sp.n.) was confirmed; case 3, the previously supposed presence of T. casta (Rondani, 1859) in central Europe was not supported; case 4, West Palaearctic T. nupta (Rondani, 1859) was contrasted with East Palaearctic specimens from Japan, which seem to represent a valid species not conspecific with central European specimens; T. nupta needs detailed further study; case 5, T. nigrohirta (Stein, 1924) resurrected recently from synonymy with T. ursina Meigen, 1824 was confirmed as a valid species. This parallel application of three alternative methods has enabled the principle of ‘polyphasic taxonomy’ to be tested and verified using these separate results. For the first time, the value of using the ANN approach in taxonomy was justified by two non-mathematical methods (molecular and morphological).

Ruminant dung is a highly ephemeral, patchily distributed resource, which is used by a diverse community of invertebrate species. In such environments, high levels of insect aggregation may be important in facilitating stability and coexistence across patchily distributed populations. The aim of the present work was to quantify the aggregation of the insects colonising cow-dung in cattle pasture in southwest England and to test the hypothesis that the dung-pat community assemblage observed was the result of stochastic colonization. This was examined using batches of ten standardised, 1.5 kg, artificial cow pats placed out in cattle pastures in each of 24 weeks between May and October in 2001. Pats were left exposed in the field for seven days before being brought back to the laboratory, where any insect colonizers were collected and identified. Individual pats contained, on average, only half the number of insect taxa present in an entire batch put out at any one time. All larval coleopteran taxa, 20 of the 22 adult coleopteran taxa and 22 of the 23 dipteran taxa, showed significant aggregation, with the abundance of most taxa within pats approximating a negative binomial distribution. A simulation analysis was used to show that the observed relative frequency of taxa within pats did not differ from that expected by chance if colonisation is a random binomial event in which each species colonises a pat independently of all other species. Aggregated populations, of even highly abundant insects, may be more susceptible to the deleterious effects of insecticidal contaminants in dung than if they were evenly distributed, if by chance they colonize a pat containing insecticidal residues from a recently treated animal.