Field- and laboratory-based studies on correlates of Chlamydia trachomatis transmission by Musca sorbens: Determinants of fly-eye contact and investigations into fly carriage of elementary bodies.

Musca sorbens (Diptera: Muscidae) flies are thought to be vectors of the blinding eye disease trachoma, carrying the bacterium Chlamydia trachomatis (Ct) between the eyes of individuals. While their role as vectors has been convincingly demonstrated via randomised controlled trials in The Gambia, studies of fly-borne trachoma transmission remain scant and as such our understanding of their ability to transmit Ct remains poor. We examined fly-eye contact and caught eye-seeking flies from 494 individuals (79% aged ≤9 years) in Oromia, Ethiopia. Ct-carrying flies (harbouring Ct DNA) were found to cluster spatially in and nearby to households in which at least one resident had Ct infection. Fly-eye contact was positively associated with the presence of trachoma (disease), lower human body weight and increased human body temperature. Studies of laboratory-reared M. sorbens indicated that Ct is found both externally and internally following feeds on Ct culture, with scanning electron microscopy revealing how Ct bodies can cling to fly hairs (setae). Testing for Ct on field-caught M. sorbens found fly 'bodies' (thorax, wings and abdomen) to consistently test positive for Ct while legs and heads were infrequently Ct-positive. These studies strongly support the role of M. sorbens as vectors of trachoma and highlight the need for improved understanding of fly-borne trachoma transmission dynamics and vector competence.

Unveiling the hidden dangers: enteropathogens carried by flies in Pudong New Area.

BACKGROUND: Flies are acknowledged as vectors of diseases transmitted through mechanical means and represent a significant risk to human health. The study aimed to determine the prevalence of enteropathogens carried by flies in Pudong New Area to inform strategies for preventing and controlling flies. METHODS: Samples were collected from various locations in the area using cage trapping techniques between April and November 2021, encompassing various habitats such as parks, residential areas, restaurants, and farmers' markets. The main fly species were identified using cryomicrography and taxonomic enumeration, with 20 samples per tube collected from different habitats. Twenty-five enteropathogens were screened using GI_Trial v3 TaqManTM microbial arrays. RESULTS: A total of 3,875 flies were collected from 6,400 placements, resulting in an average fly density of 0.61 flies per cage. M. domestica were the most common species at 39.85%, followed by L. sericata at 16.57% and B. peregrina at 13.14%. Out of 189 samples, 93 tested positive for enteropathogens, with nine different pathogens being found. 12.70% of samples exclusively had parasites, a higher percentage than those with only bacteria or viruses. The study found that M. domestica had fewer enteropathogens than L. sericata and B. peregrina, which primarily harbored B. hominis instead of bacteria and viruses such as E. coli, Astrovirus, and Sapovirus. During spring testing, all three fly species exhibited low rates of detecting enteropathogens. M. domestica were found in residential areas with the highest number of pathogen species, totaling six. In contrast, L. sericata and B. peregrina were identified in farmers' markets with the highest number of pathogen species, totaling six and seven, respectively. CONCLUSIONS: Flies have the potential to serve as vectors for the transmission of enteropathogens, thereby posing a substantial risk to public health.

An illustrated identification key to early instar larvae of forensically important Muscidae (Diptera) of the western Palaearctic region.

There is a significant gap in the availability of comprehensive identification keys for the early larval stages of forensically important fly species. While well-documented identification keys exist for the third instar larvae, particularly for the Calliphoridae, Muscidae and Sarcophagidae families, there is a notable scarcity of keys for the first, except Calliphoridae, and the second instar larvae, with no such resources available for muscid species. The second instar larvae suffer the most from the lack of morphological descriptions and available identification keys. The Muscidae is one of the most frequently reported dipteran families of forensic importance colonising animal cadavers and human corpses. Nevertheless, descriptions of the morphology of their early instars remain scarce and limited to only a few species, thus their larval identification is challenging or impossible. Considering the numerous challenges associated with studying small-sized entomological material, we tested whether it is feasible to identify muscid flies to the species or at least genus level based predominantly on the details of the cephaloskeleton. To overcome the obstacle of observing details of small sclerites, especially their shapes and interconnections, we effectively employed confocal laser scanning microscopy (CLSM) as a supplementary method for light microscopy (LM). This study provides an identification key for first and second instar larvae of forensically important muscid species from the western Palaearctic (Europe, North Africa, Middle East). The proposed key primarily utilises details of the cephaloskeleton with only addition of external morphology.

A review of Musca sorbens (Diptera: Muscidae) and Musca domestica behavior and responses to chemical and visual cues.

Musca flies (Diptera: Muscidae) have been found culpable in the mechanical transmission of several infectious agents, including viruses, bacteria, protozoans, and helminths, particularly in low-income settings in tropical regions. In large numbers, these flies can negatively impact the health of communities and their livestock through the transmission of pathogens. In some parts of the world, Musca sorbens is of particular importance because it has been linked with the transmission of trachoma, a leading cause of preventable and irreversible blindness or visual impairment caused by Chlamydia trachomatis, but the contribution these flies make to trachoma transmission has not been quantified and even less is known for other pathogens. Current tools for control and monitoring of house flies remain fairly rudimentary and have focused on the use of environmental management, insecticides, traps, and sticky papers. Given that the behaviors of flies are triggered by chemical cues from their environment, monitoring approaches may be improved by focusing on those activities that are associated with nuisance behaviors or with potential pathogen transmission, and there are opportunities to improve fly control by exploiting behaviors toward semiochemicals that act as attractants or repellents. We review current knowledge on the odor and visual cues that affect the behavior of M. sorbens and Musca domestica, with the aim of better understanding how these can be exploited to support disease monitoring and guide the development of more effective control strategies.

Black blow fly (Diptera: Calliphoridae) bacterial symbionts inform oviposition site selection by stable flies (Diptera: Muscidae).

Larval habitats of blood-feeding stable flies, Stomoxys calcitrans (L.) (Diptera: Muscidae), overlap with foraging sites of black blow flies, Phormia regina (Meigen) (Diptera: Calliphoridae). We tested the hypothesis that bacteria in blow fly excreta inform oviposition decisions by female stable flies. In laboratory 2-choice bioassays, we offered gravid female stable flies fabric-covered agar plates as oviposition sites that were kept sterile or inoculated with either a blend of 7 bacterial strains isolated from blow fly excreta (7-isolate-blend) or individual bacterial isolates from that blend. The 7-isolate-blend deterred oviposition by female stable flies, as did either of 2 strains of Morganella morganii subsp. sibonii. Conversely, Exiguobacterium sp. and Serratia marcescens each prompted oviposition by flies. The flies' oviposition decisions appear to be guided by bacteria-derived semiochemicals as the bacteria could not be physically accessed. Oviposition deterrence caused by semiochemicals of the 7-isolate-blend may help stable flies avoid competition with blow flies. The semiochemicals of bioactive bacterial strains could be developed as trap lures to attract and capture flies and deter their oviposition in select larval habitats.

Molecular detection and genotyping of Dientamoeba fragilis and Blastocystis sp. in housefly Musca domestica (Diptera: Muscidae): first report for Dientamoeba fragilis.

Dientamoeba fragilis and Blastocystis sp. are single-celled protozoan parasites of humans and animals. Although they are found in the intestines of healthy hosts, the pathogenicity of them is still unclear. To date, there is no report on D. fragilis and only two studies (without subtyping) on the occurrence of Blastocystis sp. in Musca domestica. In this study, fly samples were collected from livestock farms and their surroundings in the Kirsehir province (Central Anatolia Region) of Türkiye from May to August 2023. A total of 150 microscopically identified M. domestica samples were analyzed for the detection of D. fragilis and Blastocystis sp. molecularly. The overall prevalence of Blastocystis sp. and D. fragilis in M. domestica was determined to be 3.3% (5/150) and 8.0% (12/150), respectively. The SSU rRNA gene sequences of the isolates indicated genotype 1 of D. fragilis. Eleven isolates were identical and represented a single isolate (KAU-Dfrag1). BLAST analysis of KAU-Dfrag1 indicated identity with the isolates reported from humans, cattle, sheep, and budgerigars. The other isolate (KAU-Dfrag2) was polymorphic at two nucleotides from KAU-Dfrag1 and three nucleotides from known genotypes from GenBank and represented a variant of genotype 1. The Blastocystis sp. isolates were found to be identical and represent a single genotype (KAU-Blast1). BLAST analysis revealed that the KAU-Blast1 genotype belonged to the potentially zoonotic subtype 5 (ST5) and exhibited the highest genetic identity (ranging from 99.4 to 99.6%) with pigs, cattle, and sheep from different countries. Our study provides the first data on the molecular prevalence, epidemiology, and genotypic characterization of D. fragilis and Blastocystis sp. in M. domestica.

Transgenerational effects of pyriproxyfen in a field strain of Musca domestica L. (Diptera: Muscidae).

Musca domestica L. (Muscidae: Diptera) is a human and livestock pest especially in tropical and sub-tropical areas. Different insecticides have been used to control this pest that pose serious harmful effects on humans and the environment. The current study was planned to investigate the effects of two concentrations (LC25 and LC50) of pyriproxyfen on biological and population parameters of a field strain of M. domestica. The exposed parents (F0) and their progeny (F1) were studied to examine the transgenerational effects. The results indicated that preadult duration was higher in control (13.68 days) compared to LC50 treated individuals (12.44 days). The male and female longevity was relatively lower in the LC25 treated population i.e. 24.62 and 26.62 days, respectively. The adult pre-oviposition period (APOP) and total pre-oviposition period (TPOP) values were higher in the LC25 treated individuals than those of control. Moreover, oviposition days and fecundity were reduced in the treated individuals as compared to the control treatment. A gradual decrease in the net reproductive rate (R0) was observed (8.46-14.07 per day) while the value of R0 was significantly higher in control. The results suggested that pyriproxyfen can be effectively utilized and incorporated in the management programs of M. domestica.

Surveillance and screening of Stomoxyinae flies from Mallorca Island (Spain) reveal the absence of selected pathogens but confirm the presence of the endosymbiotic bacterium Wolbachia pipientis.

Adult brachycera biting flies can significantly impact livestock through both direct effects (reduction of food intake, disturbance, painful bites, and blood loss) and indirect effects (pathogen transmission), leading to substantial economic losses and production damage. This study aimed to assess the presence of blood-sucking flies in six mixed-animal farm environments on the island of Mallorca (Balearic Islands, Spain) by employing multiple trapping methods. Additionally, distribution maps of brachycera biting fly species recorded in Spain were created, based on data extracted thorough review of scientific literature and citizen digital databases. Investigation of several pathogens, including equine infectious anemia virus (EIAV), Anaplasmataceae bacteria, and piroplasm protozoa, was carried out using different PCR targets (18S rRNA, 16S rRNA, groESL, and tat genes). Citizen science databases and literature review corroborated the consistent distribution trend for two Stomoxyinae species, underscoring the importance of citizen collaboration as a complement to traditional entomological surveillance. Our study confirmed the presence of two biting Stomoxyinae species: the prevalent stable fly Stomoxys calcitrans across all sampled farms, and the horn fly Haematobia irritans, which turned out to be less abundant. DNA barcoding techniques validated the identification of the two species. Neither EIAV nor bacterial/protozoan pathogens were detected using the selected PCR targets in either fly species. However, Wolbachia pipientis (clustered in the supergroup A together with the only sequence of W. pipientis from the USA) was identified through PCR targeting 16S rRNA, groESL and wsp genes in all pools of H. irritans (n = 13) collected from two of the examined farms. This study represents the first attempt to investigate pathogens in Stomoxyinae biting flies in Spain. The discovery of the endosymbiotic Wolbachia organism in H. irritans represents the first record in Spain and the second from Europe. This finding holds significant implications for future research on the applications of this bacterium in biocontrol programs.

Spatiotemporal distribution of hematophagous flies (Diptera: Muscidae) on beef cattle farms in Bangkok, Thailand.

A spatiotemporal investigation of hematophagous fly prevalence was conducted over a 1-year period on 12 beef cattle farms located in major livestock areas of Bangkok, Thailand, using Vavoua traps. The survey revealed 5,018 hematophagous flies belonging to Muscidae and Tabanidae, with the 3 dominant species identified as Stomoxys calcitrans (Linnaeus) (2,354; 46.91%), Musca crassirostris Stein (1,528; 30.45%), and Haematobia exigua de Meijere (922; 18.37%). The abundance of S. calcitrans per trap per week was significantly higher during the rainy season (45.64 ±â€…14.10), followed by the cold and dry seasons (6.39 ±â€…2.16 and 3.04 ±â€…1.27, respectively). The relative abundance of S. calcitrans reached the highest apparent density per trap per day (ADT) index of 9.83 in August 2022 during the rainy season. Subsequently, there was a rapid decline, and the ADT index dropped to nearly zero in December 2022 during the cold season. This low abundance continued through the dry months from March to May 2023. The higher rainfall and relative humidity could significantly contribute to the high relative abundance of S. calcitrans. In contrast, M. crassirostris and H. exigua showed population fluctuations that were not significantly associated with seasonal changes and weather conditions. Remote sensing data and spatial regression analyses using ordinary least squares regression showed the high spatial density of S. calcitrans in the north direction of the Khlong Sam Wa district during the rainy season; it shifted toward the south in the cold and dry seasons, corresponding with rainfall.

A new species of Sumapazomyia Fogaa, Couri, Prez & de Carvalho (Diptera: Muscidae) from Colombia.

Sumapazomyia Fogaa et al. 2019, is a monotypic Andean genus from the Colombian Andes. Here we described one new species of Sumapazomyia from Colombia, namely Sumapazomyia quimbaya sp. nov. (type-locality: Risaralda, Otn, Quimbaya, Santuario de Fauna y Flora Otn Quimbaya), and an identification key to segregate the two species, including color images and illustrations of male and female terminalia is given. The distribution of the two species is mapped.

Entomological study on vectorial density, temporal variation of tsetse fly and other biting flies in intervention and non-intervention areas of South Omo Zone, Ethiopia.

A repeated cross-sectional entomological survey was conducted to estimate Glossina (tsetse) and other biting flies density, their seasonal variation and associated risk factors in intervention and non- intervention areas of South Omo Zone, Southwest Ethiopia from January 2019-November 2019. In both dry and wet seasons, a total of 96 NGU traps (64 traps in tsetse intervention districts and 32 traps in tsetse non- intervention districts) were deployed at an interval of about 100-200 m in purposively selected and suspected tsetse habitats. Thus, Glossina pallidipes was found to be the only cyclical vector along with mechanical vectors of Tabanus, Stomoxys and Haematopota. In tsetse intervention areas, G. pallidipes apparent density of 2.64 F/T/D and 0.42 F/T/D was recorded in dry and wet season respectively. Mechanical vectors (dry; wet) of Tabanus (205; 155), Stomoxys (34; 54) and Haematopota (50; 33) were also recorded in tsetse intervened areas. Whereas, in non- intervention areas, apparent density of G. pallidipes was 2.03 F/T/D and 0.56 F/T/D, respectively in dry and wet season. Similarly, Tabanus (22; 56), Stomoxys (10; 8) and Haematopota (5; 7) respectively in dry and wet (dry; wet) season were recorded in tsetse non- intervention areas. According to Negative Binomial Regression (NBR), season was the only variable significantly affecting (P < 0.05) the Glossina count in the current study area. Accordingly, the incidence G. pallidipes during wet season was decreased by the factor of 0.21 (CI; 0.097-0.47) when compared to its incidence in dry season by holding other variables constant. In conclusion, cyclical vectors were playing vital role in transmission of trypanosomosis in South Omo Zone along with numerous mechanical vectors even though there have been vector intervention activities in the areas. Therefore, strong, sustainable, environmentally friend and community participating vector control strategies should be followed to tackle the vector distribution in the area.

Using modified trapping regimes to understand the behavioral and spatial ecology of Philornis downsi (Diptera: Muscidae).

The avian vampire fly Philornis downsi (Dodge & Aitken) (Diptera: Muscidae) is native to continental South America and the Caribbean, but invasive in the Galapagos Archipelago. The larvae of P. downsi feed on the blood and tissues of the nestlings of 75% of the small land bird species that are endemic or native to Galapagos, causing high in-nest mortality and severe population declines in some species. Efficient trapping techniques are vital to safeguarding these birds in the short term as well as for monitoring fly populations, but basic information about the ecology of the fly is still needed to help develop a species-appropriate trapping method. In this study, we used a novel trapping regime with a vertical distribution to make inferences about P. downsi's behavioral and spatial ecology and to optimize trap catch. Our results showed that male and female P. downsi were trapped in greater numbers below the canopy (3.1-7.5 m), lower down than other commonly caught insect species (5.1-11.5 m). Notably, the effect of trap height remained consistent across seasons and different weather conditions. These findings suggest that P. downsi tend to move at heights where their hosts nest (at or below the canopy) and do not spend time above the canopy. This also makes it unlikely that strategies such as hill-topping or aerial swarming are being used to locate mates. As such, trapping and control efforts should be focused below the canopy in forests with similar canopy heights to effectively capture P. downsi and reduce bycatch of other insects.

Evaluation of mechanical transmission of Trypanosoma vivax by Stomoxys calcitrans in a region without a cyclic vector.

This work investigated the mechanical transmission of Trypanosoma vivax by Stomoxys calcitrans to cattle in a region without a cyclic vector. The study involved two experiments, one with calves experimentally infected with T. vivax, in the acute phase of trypanosomosis (Experiment 1) and the other in the chronic phase (Experiment 2). In both experiments, two transmission methods were used with flies that had not fed for 24 h or had never fed: (i) Method 1: flies released freely in cattle pens (≈3,300 flies/pen for 10 days); and (ii) Method 2: flies placed in a feeding chamber (12 flies/animal). To develop Method 1 in the two experiments (acute and chronic phases), T. vivax-positive animals were kept with T. vivax-negative animals. Periodically, the Brener method, Woo method, blood smears, cPCR, ELISA, IFAT, and Imunoteste® were performed to detect T. vivax in the animals. We also recorded the animals' head tossing and hoof stomping and the number of flies near the pens' inner walls. Subsequently, biological testing was performed using lambs. For Method 2 in both experiments, flies inside the feeding chamber first fed on T. vivax-positive animals and later on negative animals. In both experiments and methods, we examined the flies for the presence of T. vivax through blood smears and cPCR of the proboscis and abdomen. In Experiment 2 (chronic phase), a test was conducted to determine how long trypomastigotes forms could survive on the blood of animals with different levels of parasitemia. None of the animals (calves and lambs) became infected with T. vivax or showed antibodies against it. During the evaluation period, the animals in the presence of the flies exhibited more hoof stomping and head tossing compared to those without flies (control). Additionally, there was an increase in the number of flies in the pens during the experiment. Only in Experiment 1 (acute phase) were T. vivax trypomastigotes and DNA found in the abdomen of the flies but not in the proboscis. In Experiment 2 (chronic phase), higher concentrations of trypomastigotes per milliliter of blood were associated with a shorter the lifespan of this stage of the parasite. In conclusion, under the variable conditions of the experiments (hosts, number of flies, and level of parasitemia), S. calcitrans was unable to mechanically transmit T. vivax to cattle.

An annotated wing interferential pattern dataset of dipteran insects of medical interest for deep learning.

Several Diptera species are known to transmit pathogens of medical and veterinary interest. However, identifying these species using conventional methods can be time-consuming, labor-intensive, or expensive. A computer vision-based system that uses Wing interferential patterns (WIPs) to identify these insects could solve this problem. This study introduces a dataset for training and evaluating a recognition system for dipteran insects of medical and veterinary importance using WIPs. The dataset includes pictures of Culicidae, Calliphoridae, Muscidae, Tabanidae, Ceratopogonidae, and Psychodidae. The dataset is complemented by previously published datasets of Glossinidae and some Culicidae members. The new dataset contains 2,399 pictures of 18 genera, with each genus documented by a variable number of species and annotated as a class. The dataset covers species variation, with some genera having up to 300 samples.

Urban living can rescue Darwin's finches from the lethal effects of invasive vampire flies.

Human activity changes multiple factors in the environment, which can have positive or negative synergistic effects on organisms. However, few studies have explored the causal effects of multiple anthropogenic factors, such as urbanization and invasive species, on animals and the mechanisms that mediate these interactions. This study examines the influence of urbanization on the detrimental effect of invasive avian vampire flies (Philornis downsi) on endemic Darwin's finches in the Galápagos Islands. We experimentally manipulated nest fly abundance in urban and non-urban locations and then characterized nestling health, fledging success, diet, and gene expression patterns related to host defense. Fledging success of non-parasitized nestlings from urban (79%) and non-urban (75%) nests did not differ significantly. However, parasitized, non-urban nestlings lost more blood, and fewer nestlings survived (8%) compared to urban nestlings (50%). Stable isotopic values (δ15 N) from urban nestling feces were higher than those from non-urban nestlings, suggesting that urban nestlings are consuming more protein. δ15 N values correlated negatively with parasite abundance, which suggests that diet might influence host defenses (e.g., tolerance and resistance). Parasitized, urban nestlings differentially expressed genes within pathways associated with red blood cell production (tolerance) and pro-inflammatory response (innate immunological resistance), compared to parasitized, non-urban nestlings. In contrast, parasitized non-urban nestlings differentially expressed genes within pathways associated with immunoglobulin production (adaptive immunological resistance). Our results suggest that urban nestlings are investing more in pro-inflammatory responses to resist parasites but also recovering more blood cells to tolerate blood loss. Although non-urban nestlings are mounting an adaptive immune response, it is likely a last effort by the immune system rather than an effective defense against avian vampire flies since few nestlings survived.

Characterization of intrapuparial development of Musca domestica (Diptera: Muscidae), under different temperatures in laboratory conditions.

Musca domestica (Linnaeus, 1758) (Diptera: Muscidae), popularly known as "housefly", is a highly synanthropic species, with economic, medical-sanitary, veterinary, and forensic importance. It is able to cause damage to health, transmit pathogenic agents, cause infection in domestic animals, and, in its immature stage, cause secondary myiasis. The scavenging habit of its immature stages makes these flies pioneers in colonizing both human and animal carcasses, from the initial stages of corpse decomposition to the final stages. Intrapuparial development studies of all stages of the biological cycle of these insects help estimate pupal age, being useful to forensic entomology to aid in determining the minimum postmortem interval (minPMI). This study describes, morphologically, the external structures of the pupae, under temperatures of 23, 27, and 30 ±â€…1 °C aiming to identify the characteristics that define their developmental stages and estimation of the pupae age of M. domestica. The whole experiment was carried out under laboratory conditions, with relative humidity 60 ±â€…10% and 12 hours of photoperiod. The process of pupariation and pupation including pre-pupae phases were observed; larvae pupae apolysis; early cryptocephalic pupae; late cryptocephalic pupae; phanerocephalic pupae; pharate adult, discriminated by eye color (transparent eyes, pink eyes, and red eyes); and the emergency of adults, which occurred in the intervals of 162-180; 138-144, and 96-102 hr, respectively, being described throughout the metamorphosis of the external morphological characteristics of the pupal stage of M. domestica.

Evaluation of the stability of physiological and behavioral resistance to imidacloprid in the house fly (Musca domestica L.) (Diptera: Muscidae).

BACKGROUND: The house fly (Musca domestica L.) is a synanthropic fly species commonly associated with confined animal facilities. House fly control relies heavily on insecticide use. Neonicotinoids are currently the most widely used class of insecticide and have been formulated into granular fly baits since 2002. Physiological resistance to imidacloprid in house flies has been observed to be unstable and decline over time without continual selection pressure, indicating that resistance has a fitness cost to individuals in the absence of exposure to insecticides. The stability of behavioral resistance to imidacloprid in the house fly has not been evaluated. In the current study, we assess the stability of physiological and behavioral resistance in house flies to imidacloprid over time. RESULTS: Physiological susceptibility to imidacloprid varied significantly among three house fly strains examined, with WT-15 exhibiting the greatest susceptibility to imidacloprid with an LC50 and LC95 of 109.29 (95.96-124.49) µg g-1 and 1486.95 (1097.15-2015.23) µg g-1 , respectively. No significant differences in survival were observed across 30 generations of a house fly strain (BRS-1) previously selected for behavioral resistance to imidacloprid with percentage survival ranging from 93.20% at F0 in 2020 to 96.20% survival at F30 in 2022. CONCLUSION: These results have significant implications for the management of house flies exhibiting behavioral resistance in field settings. It appears that standard resistance management tactics deployed to reduce the prevalence of physiological resistance, such as rotating or temporarily discontinuing the use of specific insecticides, may not lead to reduced behavioral resistance to imidacloprid. © 2023 The Authors. Pest Management Science published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.

All insects matter: a review of 160 entomology cases from 1993 to 2007 in Switzerland-part I (Diptera).

Necrophagous Diptera are the most important group of insects used for the purposes of forensic entomology. While the most utilized fly family in this context is the family Calliphoridae, there are several other families that can be of great importance during real-case investigations. This article analyzes the necrophagous flies of all families recorded from 160 real cases in Switzerland between 1993 and 2007. A total of 56 species belonging to 16 families was identified with Calliphoridae being the most dominant family (90.63% of all cases), followed by Muscidae (26.25%), Sarcophagidae (19.38%), Phoridae (14.38%), and Fanniidae (12.50%). For specimens that were difficult to identify morphologically, a new PCR primer has been specifically designed for the amplification of a short, informative COI barcode in degraded museum samples of forensically important Diptera taxa. The richest family in terms of species was the family Muscidae with 16 species. Fannia fuscula (Fallen) and Fannia monilis (Haliday) were recorded from human cadavers for the first time. The study highlights the importance of different fly families in forensic investigation, enhancing our comprehension of their prevalence and dispersion in real cases in Central Europe. The results pave the way for additional exploration, especially regarding the involvement of less frequently observed species in forensic entomology.

Seasonal variation in tsetse fly apparent density and Trypanosoma spp. infection rate and occurrence of drug-resistant trypanosomes in Lambwe, Kenya.

Tsetse flies are major arthropod vectors of trypanosomes that cause debilitating African animal trypanosomiasis. The emergence of drug-resistant trypanosomes is a common problem in sub-Saharan Africa. This study aimed to identify tsetse flies' seasonal variation in apparent densities and their infection rates and the occurrence of drug-resistant trypanosomes. Tsetse flies were collected from Lambwe, Kenya, during May and September 2021. Genomic DNA was extracted from them, and the ITS1 gene was amplified to detect Trypanosoma infection with subsequent species determination. Transporter genes DMT, E6M6, TbAT/P2, and TcoAde2 were targeted to detect polymorphisms associated with drug-resistance, using sequencing and comparison to drug-sensitive trypanosome species referenced in Genbank. A total of 498 tsetse flies and 29 non-tsetse flies were collected. The apparent density of flies was higher in wet season 6.2 fly per trap per density (FTD) than in the dry season 2.3 FTD (P = 0.001), with n = 386 and n = 141 flies caught in each season, respectively. Male tsetse flies (n = 311) were more numerous than females (n = 187) (P = 0.001). Non-tsetse flies included Tabanids and Stomoxys spp. Overall, Trypanosoma infection rate in tsetse was 5% (25/498) whereby Trypanosoma vivax was 4% (11/25), Trypanosoma congolense 36% (9/25), and Trypanosoma brucei 20% (5/25) (P = 0.186 for the distribution of the species), with infections being higher in females (P = 0.019) and during the wet season (P < 0.001). Numerous polymorphisms and insertions associated with drug resistance were detected in DMT and E6M6 genes in two T. congolense isolates while some isolates lacked these genes. T. brucei lacked TbAT/P2 genes. TcoAde2 sequences in three T. congolense isolates were related to those observed in trypanosomes from cattle blood in our previous study, supporting tsetse fly involvement in transmission in the region. We report Trypanosoma associated with trypanocidal drug-resistance in tsetse flies from Lambwe, Kenya. Female tsetse flies harbored more Trypanosoma infections than males. Tsetse transmission of trypanosomes is common in Lambwe. Risk of trypanosome infection would seem higher in the wet season, when tsetse flies and Trypanosoma infections are more prevalent than during the dry season. More efforts to control animal trypanosome vectors in the region are needed, with particular focus on wet seasons.

Control study of Musca domestica (Diptera, Muscidae) in Misan Province.

Background: Houseflies are the most common type of Diptera, specifically Muscidae, worldwide, representing more than 90% of all species. This family has over 170 genera and 4200 species, but a few are of medical significance. This study aimed to estimate and assessing the measures to control and prevent grow-up inside houses and flying of the housefly ( Musca domestica Linnaeus, 1758) in Misan. Methods: The study occurred over 12 months, from December 2020 to December 2021. Using plastic containers, Musca domestica were collected from all potential breeding sites in the study region (inside and around houses). Sticky oil paper and traps were used to collect the insects. The collected insects were transferred to sealed plastic containers and then to the laboratory of the Department of Microbiology.Out of 200 randomly selected houses, 150 (75%) contained insects. Light traps and sticky oil papers were the most effective control measures, with 26.7% and 25.9% of the Musca domestica collected from these methods, respectively. The ratio of male (233) to female (456) Musca was 1:2, with a significant difference between the frequencies (P<0.05). A large population of houseflies was collected during the hot season (501, 72.7%), whereas fewer Musca were collected during the cold months (188, 27.3%), with a strongly significant difference (P<0.05). The percentage of HI was 54.4%, the CI was 21.9%, and the BI was 79.9%. The overall larval densities (LD) were at a medium level. Conclusions: Misan has a high density of Musca domestica, with females being more prominent than males. Hot climate, humid sites, and dirty places are responsible for the breeding of houseflies. The overall larval density was medium. Therefore, the risk of transmitting infectious diseases by houseflies is high within the boundaries of Misan province, and effective control parameters should include measures like light traps and sticky oil.