Post-invasion selection acts on standing genetic variation despite a severe founding bottleneck.

Invasive populations often have lower genetic diversity relative to the native-range populations from which they derive.1,2 Despite this, many biological invaders succeed in their new environments, in part due to rapid adaptation.3,4,5,6 Therefore, the role of genetic bottlenecks in constraining the adaptation of invaders is debated.7,8,9,10 Here, we use whole-genome resequencing of samples from a 10-year time-series dataset, representing the natural invasion of the Asian honey bee (Apis cerana) in Australia, to investigate natural selection occurring in the aftermath of a founding event. We find that Australia's A. cerana population was founded by as few as one colony, whose arrival was followed by a period of rapid population expansion associated with an increase of rare variants.11 The bottleneck resulted in a steep loss of overall genetic diversity, yet we nevertheless detected loci with signatures of positive selection during the first years post-invasion. When we investigated the origin of alleles under selection, we found that selection acted primarily on the variation introduced by founders and not on the variants that arose post-invasion by mutation. In all, our data highlight that selection on standing genetic variation can occur in the early years post-invasion, even where founding bottlenecks are severe.

Higher prevalence of sacbrood virus in Apis mellifera (Hymenoptera: Apidae) colonies after pollinating highbush blueberries.

Highbush blueberry pollination depends on managed honey bees (Apis mellifera) L. for adequate fruit sets; however, beekeepers have raised concerns about the poor health of colonies after pollinating this crop. Postulated causes include agrochemical exposure, nutritional deficits, and interactions with parasites and pathogens, particularly Melisococcus plutonius [(ex. White) Bailey and Collins, Lactobacillales: Enterococcaceae], the causal agent of European foulbrood disease, but other pathogens could be involved. To broadly investigate common honey bee pathogens in relation to blueberry pollination, we sampled adult honey bees from colonies at time points corresponding to before (t1), during (t2), at the end (t3), and after (t4) highbush blueberry pollination in British Columbia, Canada, across 2 years (2020 and 2021). Nine viruses, as well as M. plutonius, Vairimorpha ceranae, and V. apis [Tokarev et al., Microsporidia: Nosematidae; formerly Nosema ceranae (Fries et al.) and N. apis (Zander)], were detected by PCR and compared among colonies located near and far from blueberry fields. We found a significant interactive effect of time and blueberry proximity on the multivariate pathogen community, mainly due to differences at t4 (corresponding to ~6 wk after the beginning of the pollination period). Post hoc comparisons of pathogens in near and far groups at t4 showed that detections of sacbrood virus (SBV), which was significantly higher in the near group, not M. plutonius, was the primary driver. Further research is needed to determine if the association of SBV with highbush blueberry pollination is contributing to the health decline that beekeepers observe after pollinating this crop.

Honey bee stressor networks are complex and dependent on crop and region.

Honey bees play a major role in crop pollination but have experienced declining health throughout most of the globe. Despite decades of research on key honey bee stressors (e.g., parasitic Varroa destructor mites and viruses), researchers cannot fully explain or predict colony mortality, potentially because it is caused by exposure to multiple interacting stressors in the field. Understanding which honey bee stressors co-occur and have the potential to interact is therefore of profound importance. Here, we used the emerging field of systems theory to characterize the stressor networks found in honey bee colonies after they were placed in fields containing economically valuable crops across Canada. Honey bee stressor networks were often highly complex, with hundreds of potential interactions between stressors. Their placement in crops for the pollination season generally exposed colonies to more complex stressor networks, with an average of 23 stressors and 307 interactions. We discovered that the most influential stressors in a network-those that substantively impacted network architecture-are not currently addressed by beekeepers. Finally, the stressor networks showed substantial divergence among crop systems from different regions, which is consistent with the knowledge that some crops (e.g., highbush blueberry) are traditionally riskier to honey bees than others. Our approach sheds light on the stressor networks that honey bees encounter in the field and underscores the importance of considering interactions among stressors. Clearly, addressing and managing these issues will require solutions that are tailored to specific crops and regions and their associated stressor networks.

Environmental metagenetics unveil novel plant-pollinator interactions.

Honey bees are efficient pollinators of flowering plants, aiding in the plant reproductive cycle and acting as vehicles for evolutionary processes. Their role as agents of selection and drivers of gene flow is instrumental to the structure of plant populations, but historically, our understanding of their influence has been limited to predominantly insect-dispersed flowering species. Recent metagenetic work has provided evidence that honey bees also forage on pollen from anemophilous species, suggesting that their role as vectors for transmission of plant genetic material is not confined to groups designated as entomophilous, and leading us to ask: could honey bees act as dispersal agents for non-flowering plant taxa? Using an extensive pollen metabarcoding dataset from Canada, we discovered that honey bees may serve as dispersal agents for an array of sporophytes (Anchistea, Claytosmunda, Dryopteris, Osmunda, Osmundastrum, Equisetum) and bryophytes (Funaria, Orthotrichum, Sphagnum, Ulota). Our findings also suggest that honey bees may occasionally act as vectors for the dispersal of aquatic phototrophs, specifically Coccomyxa and Protosiphon, species of green algae. Our work has shed light on the broad resource-access patterns that guide plant-pollinator interactions and suggests that bees could act as vectors of gene flow, and potentially even agents of selection, across Plantae.

Validating a multi-locus metabarcoding approach for characterizing mixed-pollen samples.

BACKGROUND: The mutualistic interaction between entomophilous plants and pollinators is fundamental to the structure of most terrestrial ecosystems. The sensitive nature of this relationship has been disrupted by anthropogenic modifications to natural landscapes, warranting development of new methods for exploring this trophic interaction. Characterizing the composition of pollen collected by pollinators, e.g. Apis mellifera, is a common means of exploring this relationship, but traditional methods of microscopic pollen assessment are laborious and limited in their scope. The development of pollen metabarcoding as a method of rapidly characterizing the abundance and diversity of pollen within mixed samples presents a new frontier for this type of work, but metabarcoding may have limitations, and validation is warranted before any suite of primers can be confidently used in a research program. We set out to evaluate the utility of an integrative approach, using a set of established primers (ITS2 and rbcL) versus melissopalynological analysis for characterizing 27 mixed-pollen samples from agricultural sites across Canada. RESULTS: Both individual markers performed well relative to melissopalynology at the family level with decreases in the strength of correlation and linear model fits at the genus level. Integrating data from both markers together via a multi-locus approach provided the best rank-based correlation between metagenetic and melissopalynological data at both the genus (ρ = 0.659; p < 0.001) and family level (ρ = 0.830; p < 0.001). Species accumulation curves indicated that, after controlling for sampling effort, melissopalynological characterization provides similar or higher species richness estimates than either marker. The higher number of plant species discovered via the metabarcoding approach simply reflects the vastly greater sampling effort in comparison to melissopalynology. CONCLUSIONS: Pollen metabarcoding performed well at characterizing the composition of mixed pollen samples relative to a traditional melissopalynological approach. Limitations to the quantitative application of this method can be addressed by adopting a multi-locus approach that integrates information from multiple markers.

A new species in Simulium (Trichodagmia) (Diptera: Simuliidae) from Chapada Diamantina region, Brazil: cryptic diversity revealed by morphological and molecular evidence.

We describe new species of black fly that had previously been identified as S. scutistriatum Lutz due to morphological similarities at the pupal stage. The description of the new species, Simulium (Trichogamia) itajara n. sp., is based on molecular and morphological evidences. The known distribution of the new species is currently restricted to the Paraguaçu River hydrographic basin in Chapada Diamantina National Park and the surrounded area in Bahia state, Brazil. The distribution record for S. scutistriatum in the northeast region of Brazil needs to be removed, since the previous records were based on occurrence of S. itajara n. sp.

Simulium (Inaequalium) marins, a new species of black fly (Diptera: Simuliidae) from inselbergs in Brazil.

A new species of black fly, Simulium (Inaequalium) marins (Diptera: Simuliidae), is described based on the male, female, pupa and larva. This new species was collected from two localities: a small stream on the Pico dos Marins, a high mountain with granite outcrops in Piquete County, state of São Paulo, and in a small stream in the Serra dos Orgãos National Park, state of Rio de Janeiro, Brazil.

Redescription of adults of Simulium jeteri (Py-Daniel, Darwich, Mardini, Strieder amp; Coscarón, 2005) (Diptera: Simuliidae).

Simulium jeteri (Py-Daniel, Darwich, Mardini, Strieder Coscarón, 2005) was not fully described, because only pharate male and female adults extracted from pupae were used for the original descriptions. Important diagnostic characters were either incompletely or poorly described. During field work in areas at the same hydrographic basin of the type-locality, we were able to rear adult stages. The main objective of this study is to redescribe the male and female of S. jeteri, providing a complete description, including morphological characteristics not reported before, such as, presence of a median ventral projection on male ventral plate and a bilobed anal lobe on female terminalia. Additional characters of pupae and larvae are given. The addition of new characters will be useful to better understand phylogenetic relationships among species in this group. Females of S. jeteri are not known to bite.

DNA Barcodes for Forensically Important Fly Species in Brazil.

Here, we analyze 248 DNA barcode sequences of 35 fly species of forensic importance in Brazil. DNA barcoding can be effectively used for specimen identification of these species, allowing the unambiguous identification of 31 species, an overall success rate of 88%. Our results show a high rate of success for molecular identification using DNA barcoding sequences and open new perspectives for immature species identification, a subject on which limited forensic investigations exist in Tropical regions. We also address the implications of building a robust forensic DNA barcode database. A geographic bias is recognized for the COI dataset available for forensically important fly species in Brazil, with concentration of sequences from specimens collected mainly in sites located in the Cerrado, Mata Atlântica, and Pampa biomes.

DNA barcoding of odonates from the Upper Plata basin: Database creation and genetic diversity estimation.

We present a DNA barcoding study of Neotropical odonates from the Upper Plata basin, Brazil. A total of 38 species were collected in a transition region of "Cerrado" and Atlantic Forest, both regarded as biological hotspots, and 130 cytochrome c oxidase subunit I (COI) barcodes were generated for the collected specimens. The distinct gap between intraspecific (0-2%) and interspecific variation (15% and above) in COI, and resulting separation of Barcode Index Numbers (BIN), allowed for successful identification of specimens in 94% of cases. The 6% fail rate was due to a shared BIN between two separate nominal species. DNA barcoding, based on COI, thus seems to be a reliable and efficient tool for identifying Neotropical odonate specimens down to the species level. These results underscore the utility of DNA barcoding to aid specimen identification in diverse biological hotspots, areas that require urgent action regarding taxonomic surveys and biodiversity conservation.

Insights into the epidemiological link between biting flies and pemphigus foliaceus in southeastern Brazil.

BACKGROUND: Black fly and sandfly bites are related to the endemicity of pemphigus foliaceus (PF); however, an immune reaction against the salivary proteins from these flies still requires confirmation in the case of PF patients living in southeastern Brazil. PURPOSE: To georeference the distribution of Simuliidae (Diptera: Simuliidae) and Phlebotominae (Diptera: Psychodidae) and of PF cases in the northeastern region of São Paulo State, and to assess the humoral immune response against salivary gland extracts (SGEs) from biting flies in PF patients, relatives, and neighbours. METHODS: PF patients' medical information recorded between 1965 and 2014 were obtained from the database of the University Hospital. Data on the distribution of fly species were collected from scientific reports and epidemiological databases. Spatial maps relating the distribution of biting flies with PF cases were plotted. Serum IgG antibodies against the SGEs from Simulium nigrimanum, Nyssomyia neivai, and Aedes aegypti (as control) were determined by ELISA. RESULTS: Two hundred and eighty-five PF cases were distributed in 60 municipalities with a prevalence of 57.5 per million inhabitants, revealing well-defined geographical clusters. S. nigrimanum and N. neivai specimens were registered in eight (13.3%) and 26 (43.3%) of these municipalities, respectively. PF patients, and their relatives presented higher levels of IgG against the SGEs of S. nigrimanum and N. neivai (P<0.001 for both), but not against the SGE from A. aegypti (P=0.115 and P=0.552, respectively), as compared to controls. IgG against the SGEs from S. nigrimanum and N. neivai but not against the SGE from A. aegypti correlated with levels of anti-Desmoglein 1 in PF patients (r=0.3848, P=0.039; and r=0.416, P=0.022, respectively). CONCLUSION: An epidemiological link between biting flies and PF in southeastern Brazil is proposed, implying a possible role of the salivary proteins from these flies in PF etiopathogenesis.

Blow Flies from Forest Fragments Embedded in Different Land Uses: Implications for Selecting Indicators in Forensic Entomology.

Given the general expectation that forest loss can alter biodiversity patterns, we hypothesize that blow fly species abundances differ in a gradient of native vegetation cover. This study was conducted in 17 fragments across different landscapes in central Brazil. Different land cover type proportions were used to represent landscape structure. In total, 2334 specimens of nine species of Calliphoridae were collected. We used principal component analysis (PCA) to reduce dimensionality and multicollinearity of the landscape data. The first component explained 70%, and it represented a gradient of forest-pasture land uses. Alien species showed a wide distribution in different fragments with no clear relationship between the abundance values and the scores of PCA axes, whereas native species occurred only in areas with a predominance of forest cover. Our study revealed that certain native species may be sensitive to forest loss at the landscape scale, and they represent a bioindicator in forensic entomology.

A systematic study on Endotribelos Grodhaus (Diptera: Chironomidae) from Brazil including DNA barcoding to link males and females.

Six new species of Endotribelos from Brazil are described and illustrated as male, female, pupa and larva: E. bicolor sp. n., E. fulvidus sp. n., E. jaragua sp. n., E. jiboia sp. n., E. semibruneus sp. n. and E. sublettei sp. n. The female of E. calophylli Roque & Trivinho-Strixino and the larvae of four unknown morphotypes are also described. Keys including males and larvae of all known species of Endotribelos are provided. Adults' males and females from five species were linked using DNA Barcoding mtCOI sequences.

A new species of Simulium (Chirostilbia) (Diptera: Simuliidae) from Mantiqueira mountain range, southeastern Brazil.

Simulium (Chirostilbia) vitribasi n. sp. from the Mantiqueira mountain range, in São Paulo and Minas Gerais states, southeastern Brazil, is described and illustrated based on male, female, pupal and larval morphologies. The pupae of the new species is similar to the one of Simulium pertinax Kollar, one of the most voracious black fly species in the South and Southeast regions of Brazil and a target of control programs in these regions. But the pupa of the new species has a large basal fenestra in the gills that distinguishes it from all other S. (Chirostilibia) species. The other life stages of the new species also have morphological characters that distinguish it from the other species in the subgenus: male has scutum black, with patches of thick, golden hairs; female has scutum similar to that of the male, without banding or other markers, abdominal tergites I and II whitish and sternite VIII dark, heavily sclerotized; anal lobe almost as wide as long, with thin, long hairs on its outer margin; larva body cuticle has wide scale-like setae, with lobulated distal margins. Females were not observed biting humans during the fieldwork.

Black-fly assemblage distribution patterns in streams in disturbed areas in southern Brazil.

Black flies (Simuliidae) are considered to be pests in rural and in some urban areas in the south of Brazil due to the bites of females. Little information exists on the factors that contribute to Simuliidae distribution in these areas. We sampled 39 streams in the state of Rio Grande do Sul in September 2004 to address ecological aspects. Of the 18 species collected, those that occurred in more than 50% of the streams were: Simulium subnigrum Lutz 1910, Simulium pertinax Kollar 1882, Simulium subpallidum Lutz 1910 and Simulium incrustatum Lutz 1910. Species richness was not influenced by any of the environmental or geographical variables considered, and there was no pattern of species co-occurrence, corroborating the hypothesis of a stochastic distribution of the Simuliidae assemblage. When analyzed individually, only three of seven species with occurrence frequency >30% were related to any of the variables measured. The probabilities of occurrence of Simulium jujuyense Paterson & Shannon 1927 and S. pertinax were higher in streams with high values of water pH and electrical conductivity, whereas the opposite was observed for S. incrustatum. Therefore, due to stochastic distribution, the population studies of Simuliidae are more suitable for predicting the occurrence of species in rural areas than are studies of assemblages.

Species richness and distribution of blackflies (Diptera: Simuliidae) in the Chapada Diamantina region, Bahia, Brazil.

Taxonomically, blackflies (Simuliidae) are among the best-known aquatic insects in Brazil. However, information on their distribution is lacking for many regions. We sampled simuliids in 50 streams in the Chapada Diamantina region, State of Bahia, located in three distinct geographical areas, 20 at Lençóis, 18 at Mucugê, and 12 at Rio de Contas. We analyzed simuliid species richness and distribution in the Chapada Diamantina area as a whole, as well as in each of the three distinct geographical areas. We collected 20 species, three of which were not yet described and two were described after our sampling. The three areas are distinct in relation to environmental variables, mainly owing to differences in altitude and water pH. Four species were restricted to Rio de Contas, while four other species were restricted to Mucugê. Sixteen species were present in Mucugê and fifteen in Rio de Contas. Only twelve species were present in Lençóis, despite the higher sampling effort. Rio de Contas appears to be the area that harbors the highest species richness. Species richness was related to environmental factors only when evaluated on a small spatial scale (i.e. when each area was analyzed separately). Although we have not tested for causal relationships, our findings agree with other studies that showed that pH and altitude are important factors with which species richness and species distribution appear to be associated.

Checklist de Simuliidae (Insecta, Diptera) do Estado de São Paulo, Brasil / Checklist of Simuliidae (Insecta, Diptera) from São Paulo State, Brazil

Nesse artigo é fornecida uma lista de espécies de Simuliidae do Estado de São Paulo. Até o momento 52 espécies estão registradas para o estado, o que representa mais de 50% de todas as espécies de borrachudos conhecidas para o Brasil. São discutidos aspectos da riqueza de espécies no Estado de São Paulo e em diferentes regiões brasileiras.

In this paper it is provided a checklist of Simuliidae species in the State of São Paulo, Brazil. There are 52 species recorded for the state, which represents more than 50% of all black fly known species in Brazil. It is also presented data about the Simuliidae species richness in the State of São Paulo as well as in different regions of Brazil.

Larval description of Simulium (Notolepria) cuasiexiguum and Simulium (Chirostilbia) obesum and new records of black fly species (Diptera: Simuliidae) in the states of São Paulo and Minas Gerais, Brazil.

We describe the larval stage of Simulium cuasiexiguum Shelley, Luna Dias, Maia-Herzog & Lowry and Simulium obesum Vulcano and provide new distribution records of four black fly species in the State of São Paulo (Simulium cuasiexiguum, Simulium metallicum s.l. Bellardi, Simulium stellatum Gil Azevedo, Figuer6 & Maia-Herzog and Simulium oyapockense s.l. Floch & Abonnenc) and three species in the State of Minas Gerais (Simulium duodenicornium Pepinelli, Hamada & Trivinho-Strixino, Simulium obesum and Lutzsimulium pernigrum Lutz).

Distribuição de espécies de borrachudos (Diptera: Simuliidae) do Espírito Santo, Brasil / Distribution of black flies (Diptera: Simuliidae) in the State of Espírito Santo, Brazil

Entre os anos de 2004 e 2008 foram realizadas coletas de Simuliidae em várias localidades no Estado do Espírito Santo, Brasil. No total, 66 córregos e rios das doze principais bacias hidrográficas do estado foram amostrados. Dezessete espécies foram coletadas, sendo que nove delas representam ocorrências novas para o estado (Simulium pertinax, S. jujuyense, S. rubrithorax, S. subnigrum, S. travassosi, S. spinibranchium, S. hirtipupa, S. lutzianum e S. anamariae), duplicando o número de espécies conhecidas no estado.

We conducted a freshwater survey from 2004 to 2008 in several localities of Espírito Santo, Brazil. In total, 66 streams and rivers were sampled in the 12 hydrographic basins of the state. We collected and identified 17 species of which nine represent new records (Simulium pertinax, S. jujuyense, S. rubrithorax, S. subnigrum, S. travassosi, S. spinibranchium, S. hirtipupa, S. lutzianum e S. anamariae). These findings doubled the number of records of black fly species to the State of Espírito Santo, Brazil.

Simulium (Chirostilbia) bifenestratum (Diptera, Simuliidae), a new black-fly species from the Atlantic forest, State of São Paulo, Brazil.

The larva, pupa, male, and female of Simulium bifenestratum n. sp. are described and illustrated. The pupae of the new species have 10 gill filaments, thick at their base and arranged in a three-dimensional way, surrounding the head and thorax. Its pupal cocoon is peculiar, not found in any of the known Brazilian black-fly species; it is very thick and hard with two openings in the anterior region. S. bifenestratum n. sp. was collected in one stream in the Bocaina mountain chain, Atlantic forest, in São José do Barreiro county, state of São Paulo, in a high (1500 m) natural grassland. Larvae and pupae were collected on the edges of small waterfalls and in places with-high speed laminar water flow, attached to the bedrock.