Defecation by haematophagous insects allows for the rapid elimination of the water content ingested along with the blood meal from their hosts. Efficient vectors defecate during or shortly after the blood feeding, and the chances of pathogenic transmission via defecation during blood feeding to their hosts increase. The duration of blood feeding and defecation varies depending on species and sexes of haematophagous insects. Culicoides females take blood meal from their hosts. C. oxystoma Kieffer (Diptera: Ceratopogonidae) are the proven vector of many arboviruses, most importantly bluetongue viruses, and protozoa—the nematode of livestock. This study investigates the bacterial communities in defecated contents and the gut of post-defecated females of C. oxystoma. To observe any pathogenic bacteria in their defecated contents, field-collected engorged females were allowed to excrete under laboratory settings. The engorged females defecated within 48 hours of transferring onto moistened cotton beds. The defecated contents discharged by C. oxystoma consisted of the following bacteria: Bacillus cereus, Enterococcus faecium, and Alcaligenes faecalis. The following bacteria, Bacillus cereus, Lysinibacillus sp., and Paenibacillus sp., were identified from the gut of post-defecated females. Within the defecated content, the following pathogenic bacteria, B. cereus and A. faecalis, were detected, which raises serious concern and the chances of zoonotic posterior station transmission towards their hosts.

Millipedes (Diplopoda) are sexually dimorphic in various morphological, behavioral and post-embryonic development traits. Morphological intersexual variation in members of the diplopod order Julida encompasses the presence of gonopods (male specialized appendages that are involved in sperm transfer), the presence of glands that open on the coxae of the second leg-pair and modifications of gnathochilarium, mandibles and legs. In order to examine fine morphology of the latter sexually dimorphic trait in five julidan species [Cylindroiulus boleti (C. L. Koch, 1847), Megaphyllum bosniense (Verhoeff, 1897), M. unilineatum (C. L. Koch, 1838), Ommatoiulus sabulosus (Linnaeus, 1758) and Pachyiulus cattarensis (Latzel, 1884)], we dissected anterior (including the first) leg-pairs in males of the mentioned species and examined them using light and scanning electron microscopy. Our results show that the first leg-pair in all analyzed species shares common features: the legs are reduced in size (compared to other walking legs), consist of only three segments, and are hook-like. The other legs are characterized by the presence of adhesive pads on the ventral surface of the tibia and tarsi. The adhesive pads are of a ‘smooth’ type: they are made of a thin layer of cuticle with only a few striations. Since these modifications of the walking legs are present only in males of the analyzed species, we propose that these structures enable males to achieve a firmer grip of the female head (hook-like legs of the first pair) and to hold onto the female during mating (walking legs with adhesive pads).

Introduction: Determination of the synanthropic index is essential for assessing the degree of association between species, in this case, necrophagous dipterans, and humans; thus, knowledge of synanthropic relationships is crucial not only from the perspective of animal ecology and behavior but also from the standpoint of human health and hygiene.

Methods: This study aimed to detect muscid flies with the highest synanthropy and relative abundance in and around the open wet markets of the Kolkata Metropolitan Area (KMA), along with their potential risk to human health and hygiene as potential carriers of pathogenic bacteria. Furthermore, pathogenic bacteria on both the external body surface and within the gut of the most abundant eusynanthropic muscid species were assessed, highlighting potential sanitary and hygiene concerns. To detect communities of bacteria, amplicon sequencing of the V3-V4 hypervariable region (16S rDNA) was performed using the Illumina (NOVASEQ 6000) platform.

Results: The experiment was conducted from 2022 to 2023 in twelve study sites involving four municipalities. A total of 4669 muscid flies representing seven genera and thirteen species were collected using baited traps. Differences in flies’ abundance were evident across different environments. Musca domestica and Musca sorbens were the prevailing muscid species in urban and rural wet markets, with relative abundance values of 57.57 and 18.05, respectively. Musca domestica was also the most dominant fly in all of the three environments. Principal Component Analysis (PCA) indicated a robust connection between Musca domestica and urban wet markets, as evidenced by the highest synanthropic index (+91.14). Pathogenic bacterial genera like Vibrio, Aeromonas, Enterobacter, Vagococcus, Proteus, Escherichia-Shigella, Pseudomonas, Acinetobacter, Klebsiella, Morganella, etc., were identified from both the external body surface and gut of Musca domestica.

Conclusion: These results regarding muscid flies describe their role as potential vectors and can aid in assessing proper sanitation and promoting better control strategies instead of eradicating the whole fly community.

The wild cochineal Dactylopius opuntiae (Hemiptera: Dactylopiidae) is a major insect pest of the prickly pear Opuntia ficus-indica in Morocco, causing significant damage to the crop and economic losses. Chemical control is the most common method used to manage this pest. However, due to the negative impacts associated with pesticide use, plant-based pesticides represent an alternative control strategy. The present study aims to evaluate the insecticidal potential of aqueous extracts of Ricinus communis and Nicotiana glauca on D. opuntiae under field conditions. Plant extracts at 10% (w/v) concentrations were applied solely or in combination with black soap at 2% (w/v) on the nymphs and adults of D. opuntiae. Moreover, the total soluble phenol and flavonoid content in the plant extracts was determined using the spectrophotometric method with Folin–Ciocalteu and aluminum chloride (AlCl₃), respectively. The combination of each plant extract and black soap exhibited high insecticidal activity against D. opuntiae nymphs and females with 100% mortality. Regarding the individual applications, plant extracts resulted in mortality rates of 72.27 ± 12.13% and 95.94 ± 1.73% for N. glauca and R. communis extracts, respectively, while black soap showed a mortality rate of 86.42 ± 6.52%. The total phenols show a higher content in N. glauca (62.43 ± 0.59 µg GAE/mg DE) compared to R. communis (21.49 ± 0.87 µg GAE/mg DE). Meanwhile, no significant difference was recorded for flavonoid content between the two plant extracts, with 7.17 ± 0.03 µg QE/mg DE for N. glauca and 7.02 ± 0.42 µg QE/mg DE for R. communis. The findings highlight the potential of applying aqueous plant extracts combined with black soap as an eco-friendly approach for managing D. opuntiae.

Keywords: Dactylopius opuntiae; Opuntia ficus-indica L.; plant extracts; black soap; insecticidal activity

Wolbachia is considered one of the most common and widespread endosymbiotic bacteria. Wolbacha has varied impacts on the biology and evolution of its hosts. It can obscure true phylogenetic relationships and lead to erroneous mitochondrial-based phylogenetic reconstructions, species misidentification and incorrect taxonomical conclusions.

Our study aimed to shed light on the role of Wolbachia endosymbionts in the evolutionary history, biogeographic patterns and phylogenetic relationships of taxa of the genus Colias, a taxonomically challenging group of butterflies. We analyzed five nuclear (CAD, EF-1a, GADPH, RPS5, wingless) and mitochondrial (COI) genes for a large set of samples per species and conducted PCR screening for three Wolbachia genes (16S, wsp, ftsZ) in order to reveal patterns of Wolbachia infection in Colias taxa.

We discovered several cases of deeply diverged mitochondrial lineages (haplotypes) within the species that were characteristic of the species level. We found that these distinct haplotypes are usually shared by morphologically well-differentiated Colias species. Further investigation revealed the strong association of these diverged lineages with Wolbachia infection. We demonstrated that Wolbachia infection is strongly female-biased and haplotype selective: females bearing deviant haplotypes turn out to be totally infected, while males of these haplotypes, as well as males and females bearing common haplotypes, were either uninfected or had a very low rate of infection (less than 5%). We revealed that females carrying haplotypes shared by morphologically well-differentiated Colias species are always totally infected by the same Wolbachia strain. This fact suggests the transfer of Wolbachia across species through occasional hybridization resulting in mitochondrial introgression.

We conclude that Wolbachia endosymbionts play an essential role in the biology, diversification and evolution of Colias butterflies.

Financial support for this study was provided by the Ministry of Science and Higher Education of the Russian Federation (project FZMW-2023-0006).

House mice (Mus musculus L.) are known to carry ectoparasites that can cause dermatitis and serve as vectors for human diseases. However, little is known about the prevalence and diversity of ectoparasites on house mice in human apartments. In this study, we collected and identified ectoparasites from house mice trapped in apartment buildings and a chicken breeding facility across three cities in New Jersey, USA. A total of 189 house mice were captured from apartment buildings, while 113 were trapped near the chicken breeding building. Mites were the only ectoparasites detected, with the infestation rates ranging from 25% to 89% across sites. Mite species richness varied from 1.0 ± 0.0 to 2.2 ± 0.1 species per mouse. Four mite species were identified, Myocoptes musculinus (Koch), Myobia murismusculi (Schrank), Echinonyssus butantanensis (Fonseca), and Liponyssoides sanguineus (Hirst), also known as the house mouse mite. In addition, two blood-sucking mite species were genetically barcoded to support future species identification. The house mouse mite was the only medically significant species, found exclusively in apartment buildings, with a prevalence of 30% to 79% and a mean density of 2.6 ± 0.6 to 8.5 ± 1.2 mites per mouse. This study marks the first reports of E. butantanensis and the house mouse mite on house mice in New Jersey. These findings are significant, as the house mouse mite poses a potential health risk to humans in buildings where infestations are common.

Delimiting species remains a major challenge in taxonomy, particularly in groups with a minute size and high morphological similarity, such as the hyperparasitoid wasps of the subfamily Charipinae (Hymenoptera: Cynipoidea: Figitidae). Accurately selecting morphological characteristics is very important for distinguishing species, yet many traditional diagnostic traits may be unreliable due to high intraspecific variability. In this study, we applied an integrative taxonomy approach to assess species boundaries in ten species of the Alloxysta (A. brevis, A. castanea, A. obscurata, A pilipennis, A. ramulifera and A. victrix) and Phaenoglyphis (P. heterocera, P. longicornis, P. villosa and P. xanthochroa) genera. We critically evaluated morphological characteristics traditionally used in Charipinae taxonomy and tested their reliability by comparing them with molecular data from three genetic markers: mitochondrial COI, nuclear ITS2, and ribosomal 16S. Phylogenetic analyses confirmed that charipine species delineation based on morphological features is usually correct. Our results underscore the importance of integrating molecular and morphological evidence to refine species boundaries and improve the classification of hyperparasitoid wasps. By establishing a more robust framework for species identification, this study not only enhances our understanding of biodiversity within the Charipinae taxonomy but also provides essential insights for future ecological and evolutionary research involving these intricate parasitic relationships.

Lymantria mathura (Moore, 1865), the rosy gypsy moth, is a polyphagous defoliator from the family Erebidae, which inhabits deciduous forests mainly in South and Southeast Asia. The broad trophic spectrum of this species makes it an important pest that threatens forests, urban greenery, ornamental plants, and fruit trees. Due to climate change, this species could become a threat beyond its natural range and the aim of our research was to assess the risk of invasion and establishment of L. mathura in Ukraine and to model the potential distribution of natural enemies of this moth.

GIS modeling in MapInfo Pro 15.0 (ESTIMap) and IDRISI Selva (Clark Labs) was used to model the potential distribution of L. mathura, and MaxEnt was used to model the potential distribution of entomophagous insects known to be effective in L. mathura biocontrol.

We found that the potential range of L. mathura covers almost all of Ukraine, except for the Carpathian highlands. The most important bioclimatic factors influencing its establishment are the mean annual temperature, the temperature of the warmest month, and the sum of active temperatures above 10 ℃, while the limiting factor is the mean temperature of the coldest month. Modeling suggests that natural enemies such as Cotesia melanoscela (Hymenoptera: Braconidae), Compsilura concinnata, and Carcelia gnava (Diptera: Tachinidae) could play a crucial role in biological control if L. mathura invades Ukraine.

Due to its high risk of introduction, broad climatic tolerance, excellent flight ability, and numerous suitable host plants, L. mathura poses a serious threat to Ukraine's plant resources. A key risk management strategy could be its inclusion in List A1 (absent) of the Ukrainian Regulated Pests List.

The hymenopteran fauna of Bulgaria is particularly diverse, with one of the main factors being its key location on the border between the Mediterranean and Central Europe. As a result, this country is an important corridor for the migration of many species, especially along natural highways like river valleys and the Black Sea Coast.

Ongoing large-scale monitoring of these corridors, including various sampling methods combined with reviews of observations on citizen science platforms, has resulted in numerous discoveries of generally rare or previously unknown local animal species.

Here, we provide the first documented records of the digger wasp Oryttus concinnus (Rossi, 1790) and the genus Oryttus from Bulgaria, based on six specimens collected over the past three years on the territory of the capital city of Sofia (SW Bulgaria) and photographically documented observations from 2018 in Varna on the Black Sea coast and from 2025 in Sofia.

The potter wasp Rhynchium oculatum (Fabricius, 1781) has a mostly Mediterranean distribution. It was first reported in Bulgaria in Petrich (SW Bulgaria, Struma River Valley) more than 60 years ago. Here, we present a new observation of the species from Rezovo (SE Bulgaria) on the Black Sea coast, near the border of Turkey.

All Bulgarian records of Oryttus concinnus are from towns, supporting the thesis that the open urban areas are particularly suitable for digger wasps and bees because of the warmer microclimate they sustain. The observations of both species show the potential of citizen science groups for the monitoring of changes in the distribution of key species, which could complement traditional research methods tracking the impact of climate change on species distributions.

This study is a part of the project under Grant contract number KP-06-N61/6 – 14.12.2022.

The Greater melon fly, Dacus frontalis, is an economically damaging pest of cucurbit fruits in Libya. The ability of Dacus frontalis flies emerging from soil inoculated with MET52 Granular biopesticide, based on a pathogenic fungus, Metarhizium anisopliae, var anisopliae, to induce new infection in untreated flies was investigated. Contaminated adults were able to transmit the fungus conidia to untreated individuals of the opposite sex, resulting in above 30% mortality among females and 15% among males. The impact of MET52 on adult reproductive success in different mating combinations was assessed. The progeny pupation rates were affected by the treatments. The lowest pupal number was produced when inoculated males were paired with untreated females, resulting in a nearly 89% reduction. The persistence of MET52 was assessed in terms of infectivity against larval–pupal stages (measured by calculating adult emergence rate) and adult D. frontalis under laboratory conditions. The fungus reduced the emergence rate and caused mortality in newly emerging adults for more than two months after a single application.

Overall, the MET52 fungus was able to persist in soil, reducing the adult emergence rate and subsequent fly population for more than two months after a single application. In addition, it induced new infections among the fly population and reduced adult reproduction.