Thelaziasis, also known as thelaziosis, is a neglected vector-borne disease caused by parasitic nematodes from the genus Thelazia, belonging to the family Thelaziidae. The disease is transmitted biologically by various fly species, including the genus Musca (family Muscidae), Phortica (family Drosophilidae), and Fannia (family Fanniidae). These flies act as intermediate hosts (IHs) by depositing infective third-stage larvae (L3) during their tear feeding. Thelazia spp., such as T. gulosa, T. skrjabini, and T. rhodesi, inhabit the ocular regions of various animals, including humans and birds. Adult Thelazia worms are slender and creamy-white, measuring between 8-12 mm in males and 12-21 mm in females, with females releasing live larvae into the host's tear secretions. The presence of these parasites can lead to serious ocular conditions such as keratitis, conjunctivitis, and even blindness. Diagnosis typically involves identifying the adult nematodes in the conjunctival sac, where they appear as active, slender, creamy-white organisms. Management of the infection can include the physical removal of the parasites and the use of effective anthelmintics like Eprinomectin, Doramectin, Ivermectin and Levamisole, administered either subcutaneously or intramuscularly. Regular deworming and treatment of infected animals are essential practices in areas where the disease is endemic, and effective prevention and control strategies are necessary to mitigate its impact. Hence, this comprehensive review has provided updated information about the oldest known but neglected and re-emerging parasitic disease in bovines worldwide.

Fresh approach to the position of the genus Lispe Latreille in the family Muscidae (Diptera) based on the structure of pregenital and genital sclerites and the musculature of the male terminalia.

Stomoxys is a genus of blood-sucking dipteran flies from the family Muscidae with approximately 18 species reported globally. This study sought to identify and determine the apparent densities (ADs) and species richness of Stomoxys species occurring in three land-use setups, namely communal farming areas, commercial farms, and private game farms in the north-eastern part of KwaZulu-Natal Province (KZN), South Africa. Thirty-four H-traps were set up across 10 different localities over 30 days of sampling. A total of 1306 Stomoxys flies with an average of 1.28 flies/trap/day were captured, and six Stomoxys species were identified. S. n. niger was the most abundant species (82.3%), followed by S. calcitrans (13.1%), S. taeniatus (1.9%), S. n. bilineatus (0.84%), S. sitiens (1.1%), and S. boueti (0.7%) was the least collected. This study highlights the need to explore this genus further as it demonstrates that more than one species exists in north-eastern KZN. Furthermore, these flies co-exist with tsetse flies (Glossina spp.), meaning that vector control measures should incorporate all potential vectors of animal trypanosomosis and other vector-borne diseases that occur in the area.

In forensic science, information regarding identity and time of death is crucial in the legal investigation process. Traditionally, investigators determine the time of death by studying physical, chemical, histological, biochemical, and enzymatic changes that occur in a corpse. However, results obtained through these methods are only relevant within 72 hours after death. One method that can be used for a longer time frame is the observation of insects on the corpse. This study aims to observe the pattern of insect arrival on rabbit carcasses (Oryctolagus cuniculus) with different causes of death. The treatments involved complete and incomplete blood drainage, with the carcasses placed indoors under two conditions: slaughtered and lethally anesthetized. Insect observations were conducted in the morning, noon, and afternoon. The insects that arrived were then grouped according to the decomposition stages. The results showed that insects arriving during the early death to bloating stages belonged to the order Diptera (families Muscidae, Calliphoridae, Culicidae, and Sarcophagidae) and the order Hymenoptera. Specific insects that appeared during the decay stage were from the order Diptera (families Fannidae, Drosophilidae, and Asilidae). In the post-decay stage, the insects observed were from the order Diptera (families Muscidae, Calliphoridae, and Culicidae) and the order Hymenoptera.

Flowering time in Alaskan boreal forest is advancing, and this may affect pollination rates of early-flowering species. Viburnum edule is one of the first understory plants to flower, when pollinator diversity and abundance are likely lower than later in the season. We evaluated the impact of flowering time on pollen deposition and composition of the pollinator community over two years (one in which plants flowered slightly earlier than average and one in which flowering time was close to average) using experimental arrays with branches that flowered either at the start or the peak of flowering for each year. Pollinator exclusion reduced fruit set by > 90%, but even plants freely pollinated by insects had fruit set rates of < 10%. Both within and across years, plants that flowered later had more insect visitors and higher proportions of stigmas visited (> 5 pollen grains per stigma); in the advanced year, plants that flowered later also had more pollen grains per stigma. Syrphid flies, solitary bees, and muscid flies constituted ~ 99% of visitors, with a higher proportion of syrphid flies for later-flowering plants within and across years. Despite evidence for potential pollen limitation, pollen loads for the earliest flowering plants were high (mean > 25 pollen grains per stigma). Fruit production in V. edule is likely limited by inefficient pollen transfer between genets, by resource availability, or both. Given the prevalence of syrphid and muscid flies as pollinators, we need a better understanding of what triggers emergence in these taxa to evaluate the potential for trophic mismatches in boreal forest.

Introduction: Houseflies (Musca domestica L.) of the insect order Diptera and family Muscidae are found worldwide and live near humans. They are associated with dirty environments and are responsible for the transmission of pathogen scores, which remain major causative agents of health-challenging diseases in humans and livestock. This study evaluated the efficacy of Eryngium foetidum L. essential oil against adult houseflies. Methodology: The test oil was obtained via hydrodistillation using a Clevenger apparatus. From 1 ml stock solution, 15.625, 31.25, 62.50, 125, 250, and 500 µl/ml concentrations of E. foetidum oil were obtained by serial dilution using 10% Tween 80. The control consisted of 1 ml of 10% Tween 80. Both the test and control experiments were replicated three times, and 25 adult houseflies were exposed to each setup for an exposure period of 75 min. Results: Oil concentrations of 125, 250, and 500 µl/ml elicited 100% knockdown and mortality from 30 to 75 min. Houseflies exposed to the controls were neither knocked down nor killed during the duration of the experiment. The knockdown time and median lethal concentrations (KDT50 and LC50 ) of E. foetidum oil were 79.935 min and 318.758 min, respectively, as determined using the Log-time Probit Model. Conclusion: E. foetidum should therefore be further explored since it holds promise as a potent control agent against the mechanical vector, M. domestica.

Stomoxys flies are common blood-feeding pests on dairy farms and are suspected carriers of pathogenic bacteria due to their close association with manure and cattle hosts. While prior studies have used amplicon sequencing and culture-dependent methodologies to characterize the composition of the Stomoxys microbiota, little is known about strain-level acquisition of mastitis-causing bacteria from manure by Stomoxys or the functional diversity of Stomoxys-associated taxa. In this study, we address these key knowledge gaps by using whole genome sequencing to provide the first comparative genomic analysis of Stomoxys-derived Escherichia coli, Klebsiella pneumoniae, and Staphylococcaceae isolates. Our results show that fly and manure isolates collected from the same farm system are phylogenetically interspersed, with subsequent pairwise genome alignments revealing near-identical strains and plasmids shared between the two sources. We further identify a phylogenetic clade of Mammaliicoccus sciuri containing known mastitis agents associated with both flies and manure. Functional analysis reveals that this clade is highly enriched in xylose metabolism genes that are rare across other M. sciuri lineages, suggesting potential niche differentiation within the genus. Collectively, our results provide strong evidence for the acquisition of fecal-associated bacteria by adult Stomoxys flies, confirming the link between biting muscid flies and manure habitats. The intermixing of fly and manure isolates in clinically relevant taxonomic groups strongly suggests that flies serve as carriers of opportunistic mastitis-causing or other fecal-borne pathogens and may serve as important vehicles of pathogen dissemination across the dairy farm environment.IMPORTANCEBovine mastitis causes up to $32 billion dollars in losses annually in the global dairy industry. Opportunistic intramammary pathogens can be transmitted through incidental contact with bacteria in environmental reservoirs like manure. However, factors affecting the abundance, persistence, and spread of these bacteria are not well understood. Our research shows that mastitis pathogens are present in the guts of blood-feeding Stomoxys (stable) flies, which develop in cow feces and bite cows. Genomic analysis of isolates from flies, manure, and mastitis cases reveals that strains and antimicrobial resistance genes are shared between these sources. Further analysis of fly gut isolates shows virulence factors and possible niche specialization, identifying fly-associated clades with known mastitis agents from mastitic cows. This strongly suggests that Stomoxys flies play a role in the carriage and circulation of bovine mastitis pathogens from manure in dairy settings.

Antennal and palpal sensilla of two less-studied hematophagous muscid flies (Diptera: Muscidae) of veterinary significance: An ultrastructural perspective

Insects are experiencing notable phenological shifts owing to climate change, with substantial interspecific variability. However, our understanding is limited by a shortage of long-term studies, beyond Lepidoptera. This study presents a hierarchical modelling framework to analyse the phenological distribution of 11 muscid fly species across three vegetation types over 18 years (1996-2014) in Zackenberg, Northeast Greenland. We examined species-specific changes in phenology and assessed ecological traits for explaining interspecific variation. Additionally, we investigated the associations between phenological shifts and timing of snowmelt and temperature. We found consistent trends of earlier flight activity and interspecific variation in responses, with smaller species shifting their end-of-the-season activity at faster rates than larger species. Flight activity was strongly associated with the timing of snowmelt, while warming was linked to an earlier end-of-the-season activity. Late-active species exhibited more pronounced shifts in response to climate variations than early-active species. This study highlights that the species-specific climate sensitivity of high-Arctic muscid flies potentially has demographic effects if temporal overlap among interacting species changes. We advocate for prioritizing species-specific insect population studies, ideally analysed within the context of interacting species, to understand better and address disparities in responses to climate change.

Dairy farms are major reservoirs of zoonotic bacterial pathogens, which harbor antimicrobial resistance genes (ARGs), and raise critical questions about their dissemination on and off the farm environment. Here, we investigated the role of coprophagous muscid flies (Diptera: Muscidae) as carriers of zoonotic pathogens and antimicrobial resistance. We collected cow manure and flies on a dairy farm and used shotgun metagenomics to identify the presence of clinically relevant bacteria, virulence factors, and ARGs in both environments. Our results reveal that, although the fly microbiome is largely composed of manure-associated taxa, they also harbor specific insect-associated bacteria, which may be involved in nutrient provisioning to the host. Furthermore, we identifed shared ARGs, virulence factors, and zoonotic pathogens enriched within the fly gastrointestinal tract (GIT). Our study illustrates the potential flow of pathogenic microorganisms from manure to coprophagous flies, suggesting that flies may pose an important zoonotic threat on dairy farms.

We present a genome assembly from a female Phaonia tiefii (muscid fly; Arthropoda; Insecta; Diptera; Muscidae). The genome sequence has a total length of 1,537.95 megabases. Most of the assembly (99.74%) is scaffolded into 5 chromosomal pseudomolecules. The mitochondrial genome has also been assembled, with a length of 20.54 kilobases.

ABSTRACTPollinator insects are essential for the functioning of terrestrial ecosystems and play a critical role in ensuring food security; thus, studies on this topic provide significant benefits for human society. Pollination involves diverse taxa that can vary depending on the specific environment. Nevertheless, it is not always possible to monitor and study all pollinators of an ecosystem, as the research context is often constrained by financial and personnel limitations. European and national directives mostly focus on the monitoring of three taxa: bees (Hymenoptera: Apoidea: Anthophila), hoverflies (Diptera: Syrphidae) and lepidopterans (Lepidoptera). However, an effective prioritisation of taxa for pollinator research should consider their actual role as pollinators, which can vary according to the considered ecosystem. Moreover, knowledge about the specific potential of different taxa as pollinators in different ecosystems is still limited. Here, we evaluated, in mountain environments in the European Alps, the potential as pollinators of the three aforementioned taxa together with three other occurring flower‐visiting taxa: beetles (Coleoptera), muscid flies (Diptera: Muscidae) and ants (Hymenoptera: Formicidae). We compared them in terms of flower visitor abundance and actual pollen transport. In addition, we evaluated whether the scientific efforts made thus far by European researchers aligned with the actual potential of the different taxa as pollinators in a mountain context. Bees and hoverflies have been found to be both the most effective pollen carriers and the most studied pollinators. Instead, a clear discrepancy is observed for muscid flies and lepidopterans, with the former being underrepresented in the scientific literature compared to their actual potential as pollinators, while the latter are overrepresented, ranking as the second most studied taxon despite being the least abundant visitors and contributing negligibly to pollen transport. Our purpose is not to generalise our findings, but to highlight the relevant discrepancies that may exist between the actual role of pollinators in specific ecosystems and the research efforts directed towards them.

BackgroundSynanthropic filth flies thrive in human and animal habitats, posing health risks through the transmission of infectious agents. They breed on organic waste, including animal feces, making them carriers of various pathogens. In Egypt, where livestock farming is common and poor sanitation, these flies may contribute to zoonotic disease transmission. The current study investigates parasitic infections in filth flies from three livestock farms in Assiut Governorate, Upper Egypt, highlighting their role as vectors for zoonotic infections, particularly Cryptosporidium, via morphological and molecular tools.MethodsA total of 12,749 flies were collected from the study sites via sweep nets. After taxonomic identification, the flies were examined microscopically for parasites using various concentration and staining techniques. Positive samples were further confirmed for infections, particularly for Cryptosporidium parasites, via nested PCR and sequence analysis targeting the COWP and SSU rRNA genes.ResultsThis study revealed the presence of several fly species from seven dipteran families, particularly the family Muscidae, primarily Musca domestica, which presented a high parasite infestation rate of 96.6%. This study revealed a high prevalence of various protozoans and helminths in the collected flies. Cryptosporidium was the most prevalent parasite (64.4–100%), infecting all fly species. Entamoeba and Balantidium were also significant, especially in M. domestica (22.6–90.1%, 8.9–100%), Fannia canicularis (10.5–74.4%, 44.2–88.2%), and Borborillus vitripennis (11.1–50%, 37.2–91.4%). Giardia, Trichuris, and Trichostrongylidae had low to moderate prevalence in multiple fly species. Mites are commonly detected on fly exoskeletons, with high infestation rates observed in Musca domestica (77–100%) and Physiphora alceae (66.7–100%). The present study also reported sporadic infections with Trichomonas, Toxocara vitulorum, and pseudoscorpions, along with notable midge larval infestations (52.1%), mainly at site B. Parasitic infections were highest in autumn and spring, with the lowest rates in winter. Molecular identification confirmed the presence of the zoonotic species Cryptosporidium parvum and Cladotanytarsus gedanicus.ConclusionThis study revealed that zoonotic parasites exist in flies and pose potential risks when they are found near humans. Cryptosporidium parvum is the prevalent parasite causing diarrhea outbreaks in animals. This is the first genetic evidence of Cladotanytarsus gedanicus midge from Upper Egypt.

Unravelling phylogenetic relationships within the genus Lispe (Diptera: Muscidae) through genome-assisted and de novo analyses of RAD-seq data.

Muscid flies are significant in forensics and medical fields because of their polyphagous habit and capacity to spread fatal pathogens to humans and animals, causing various diseases. Adult muscids and their larvae found on carcasses and cadavers are utilized as forensic indicators to calculate the post-mortem interval (PMI). Taxonomic identification of muscid flies is difficult due to the absence of proper taxonomic expertise for quick identification of both adults and larvae. Hence, entomologists are increasingly concentrating on alternative strategies to expedite identification procedures. A short (∼670 bp) mitochondrial cytochrome oxidase I (COI) gene fragment is the optimal marker for accurate molecular identification and phylogenetic analyses. During the present study, the effectiveness of the COI gene was assessed in the molecular identification of 14 species of muscid flies in 5 genera, namely Atherigona, Hydrotaea, Musca, Neomyia, and Synthesiomyia, collected from various biogeographic regions of West Bengal. Thirty-one sequences (∼670 bp) of 14 species were generated and submitted to GenBank. The sequences of the studied species showed 99%–100% similarity with sequences from within and outside the country. COI barcodes were able to distinguish between species, with Kimura-2-Parameter (K2P) intraspecific genetic divergences ranging from 0% to 2% and Kimura-2-Parameter (K2P) interspecific genetic divergences ranging from 5.35% to 18.10%. Phylogenetic analyses involving neighbor-joining (NJ), maximum likelihood (ML), and Bayesian inference (BI) showed well-supported monophyly and proper differentiation among species. It can be concluded that COI barcodes have proven to be a highly successful alternative for identifying forensically and synanthropically significant muscid flies.

Seasonal dynamics and temporal distribution of biting and non-biting muscid flies in various zones of Karnataka state

ABSTRACTShort‐tongued saprophilous flies are a diverse but under‐appreciated group of pollinators, which are particularly important for flowers in the subtribe Stapeliinae (Apocynaceae‐Asclepiadoideae‐Ceropegieae). This clade of plants is characterised by repeated shifts between tubular kettle‐trap and open non‐trap flowers and chemical mimicry (of decomposing substrates or dying insects) to attract specific fly pollinators. The biology of most Stapeliinae, particularly those with non‐trap flowers, remains poorly studied, hampering our understanding of the mechanisms driving diversification in this plant group. We examined the pollination biology and floral traits of Ceropegia pulchellior, a South African endemic with non‐trap flowers confined to Natal Group Sandstone on the Durban escarpment. Observations showed that flowers are visited exclusively by flies which forage on small amounts of nectar in the corona cavities. Flowers were pollinated primarily by lauxaniid flies in the genus Cestrotus, although several muscid fly species also contribute. Pollinator exclusion experiments confirmed that the plants depend on these flies for reproduction. Reproductive success was low (fruit set never exceeding 8% of flowers). Pollen transfer efficiency was relatively high but variable across the flowering period. GC–MS analysis of floral scent revealed that the foetid odour is dominated by aliphatic acids and p‐cresol with small amounts of indole, supporting the assumption that C. pulchellior mimics decaying substrates to selectively attract detritus‐feeding (saprophagous) flies as pollinators. Analysis of spectral reflectance of flowers indicates that flower colours, when viewed by the fly pollinators, are not chromatically distinct from the habitat background, suggesting that flowers rely on olfactory rather than visual signals to attract pollinators. This study contributes to the growing awareness of the complexity of pollination systems in the Stapeliinae, in terms of the wide diversity of fly taxa involved, the intricate pollen transfer mechanisms, and the unusual floral scent chemistry associated with mimicry of oviposition substrates and food sources.

We present a genome assembly from an individual female muscid fly, Muscina levida (Arthropoda; Insecta; Diptera; Muscidae). The genome sequence spans 805.20 megabases. Most of the assembly is scaffolded into 5 chromosomal pseudomolecules. The mitochondrial genome has also been assembled and is 19.56 kilobases in length. Gene annotation of this assembly on Ensembl identified 13,013 protein-coding genes.

The transition of food systems for healthy diets and sustainable food systems calls for greater production of fruit and nuts, i.e. pollinated crops. The North China Plain (NCP) is a cereal dominated area that needs to transition to a more sustainable food system with a nutritionally more diverse output. Buckwheat pairs high nutritional quality with low input requirement, but it is pollination dependent. There is little knowledge on the pollinator communities and pollination service in the NCP. We selected Quzhou county as a study region on the NCP, and used potted plants of buckwheat (Fagopyrum esculentum M.) to measure pollination in three treatments: insect exclusion, open pollination, and honeybee supplementation. Measurements were made at 24 sites across the county in 2021 and 2022. At each site, we directly observed the activity of flower-visiting insects and measured the pollination rate, seed set, seed abortion rate and yield-related indicators. Most flower visits were made by hoverflies (Syrphidae) and flies from the families Muscidae, Sarcophagidae, and Calliphoridae. Compared with open pollination, insect exclusion reduced buckwheat pollination rate and seed set by more than 90 %. Without honeybee supplementation, the pollination rate in different experimental sessions was reduced by 12–21 %, and the seed set was reduced by 22–33 %. Honeybee supplementation increased harvest index and seed weight per plant with factors of 1.6 and 2.0, respectively. Hence, a pollination deficit would exist if only natural pollinators were relied upon. Managed honeybees provide an efficient means for enhancing pollination of buckwheat in the North China Plain.

Some dipteran flies play an important role in the transmission of pathogens such as viruses, bacteria, fungi, protozoan and metazoan parasites in humans and other animals. Despite this importance, knowledge of the prevalence and molecular characteristics of some pathogens in flies is limited, and no data are available for Türkiye. In this study, we investigated the possible vector role of muscid fly species for the transmission of Enterocytozoon bieneusi Desportes (Chytridiopsida: Enterocytozoonidae), Encephalitozoon spp., Coxiella burnetii Derrick (Legionellales: Coxiellaceae) and Thelazia spp. using polymerase chain reaction (PCR) and sequence analysis. The flies were trapped in different animal-related places and surroundings from two different geographical regions of Türkiye including Central Anatolia and Middle Black Sea. According to the morphological keys, 850 (85%), 141 (14.1%) and 6 (0.6%) of the total of 1000 fly specimens identified as Musca domestica Linnaeus (Diptera: Muscidae), Stomoxys calcitrans Linnaeus (Diptera: Muscidae) and Musca autumnalis De Geer (Diptera: Muscidae), respectively. The other species including Haematobia irritans Linnaeus (Diptera: Muscidae), Muscina stabulans Fallén (Diptera: Muscidae) and Hydrotaea ignava Harris (Diptera: Muscidae) were each represented by a single specimen. Screening of the pathogens identified E. bieneusi only in M. domestica with a prevalence of 2.4%. Sequence analyses identified three known genotypes, Type IV, BEB6 and BEB8, and one novel genotype named AEUEb of E. bieneusi in M. domestica. Coxiella burnetii was detected in M. domestica and S. calcitrans with prevalences of 2.9% and 2.8%, respectively. The one specimen of H. ignava was also positive for C. burnetii. Encephalitozoon spp. and Thelazia spp. were not found in the examined specimens. Our results contribute to the current knowledge on the vector potential of muscid flies and their possible role in the transmission dynamics of certain pathogens, especially in regions where diseases are prevalent and affect public and animal health.