Search

Purpouse: This study aimed to assess the influence of the presence of synanthropic flies in food preparation environments on the transmission of potentially zoonotic gastrointestinal protozoa., Methods: Flies were captured using a glass containing water, fruits, and pieces of protein., Results: Approximately 260 flies from four different species were captured: Musca domestica (76.92%; 200/260), Cochliomyia hominivorax (17.31%; 45/260), Lucilia cuprina (3.85%; 10/260), and Hydrotaea aenescens (1.92%; 5/260). Protozoa were identified through microscopic analysis after macerating these arthropods contained in their respective eppendorfs, using two techniques, namely Mini-FLOTAC ® and centrifugal sedimentation with Ziehl-Neelsen staining. The analysis revealed a concerning scenario regarding the epidemiological chain of gastrointestinal protozoa in Sergipe, Northeast Brazil. The positivity rate was 26.92% (14/52) of the pools, with 100.00% (5/5) in Aracaju, 30.00% (3/10) in Nossa Senhora do Socorro, 27.27% (3/11) in Nossa Senhora da Glória, 12.50% (2/16) in Carmópolis, and 10.00% (1/10) in Nossa Senhora das Dores. Two species of protozoa were identified, namely Cryptosporidium spp. (23.08%; 12/52) and Entamoeba spp. (9.62%; 5/52). The involvement of two fly species in the maintenance of the life cycle of these protozoa was noticeable, specifically M. domestica (27.50%; 11/40) and C. hominivorax (22.22%; 2/9)., Conclusions: Counties analyzed had an human development index (HDI) considered medium; however, they still faced socioeconomic problems such as absence of sanitation systems, waste accumulation in the streets, and illegal waste disposal, which could favor the proliferation of these vectors and the spread of these gastrointestinal protozoa., (© 2024. The Author(s), under exclusive licence to Springer Nature Switzerland AG.)

Plasmid-mediated tigecycline resistance genes, notably the tet(X) and tmexCD-toprJ genes, have garnered considerable attention due to their transferability. This study aims to investigate the prevalence and resistance mechanisms associated with tet(X) and tmexCD-toprJ in flies, which are important reservoirs of antimicrobial resistance genes. A total of 52 tigecycline resistant bacterial isolates were collected, among which 40 (76.9 %) and 12 (23.1 %) were positive for tet(X) and tmexCD-toprJ, respectively. Tigecycline resistant bacteria were isolated from diverse geographical locations in China, with tet(X4)-positive Escherichia coli and tmexCD1-toprJ1-positive Klebsiella pneumoniae dominant among the isolates. The prevalence of tet(X) in rural area was significantly higher than that in urban area (2.7 % vs. 0.3 %; P < 0.001), while the prevalence of tmexCD1-toprJ1 shows no significant difference between urban and rural areas (0.2 % vs. 0.6 %; P > 0.05). Most tet(X)-positive strains (n = 40, 100.0 %), and 11(91.7 %) of the tmexCD1-toprJ1-positive strains exhibited multi-drug resistance. The IncFIB(Mar)/IncHI1B hybrid plasmid carrying tmexCD1-toprJ1 was identified by whole-genome sequencing analysis, which dominated the transmission of tmexCD1-toprJ1 in K. pneumoniae. Genetic context analysis showed that tmexCD1-toprJ1 was related locally to IS26, and IS26 may exacerbate the spread of tmexCD1-toprJ1 in different bacteria. In addition, the genetic structure of tmexCD1-toprJ1 also contains several antimicrobial resistance genes, including aph(3')-Ic, sul1, bla DHA-1 , bla CTX-M-5 , etc., conferring resistance to aminoglycosides, sulfonamides, and carbapenems. This study provides insights into the epidemiology and transmission dynamics of tigecycline resistance genes, informing targeted intervention strategies to mitigate antimicrobial resistance dissemination., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024. Published by Elsevier Ltd.)

In flies (Diptera), the ovary displays several distinct patterns of the follicular epithelium formation and diversification. Two main patterns have been identified in the true flies or Brachycera, namely the Rhagio type and the Drosophila type. These patterns align with the traditional division of Brachycera into Orthorrhapha and Cyclorrhapha. However, studies of the follicular epithelium morphogenesis in cyclorrhaphans other than Drosophila are scarce. We characterise the developmental changes associated with the emergence of follicle cell (FC) diversity in two cyclorrhaphans belonging to the family Tephritidae (Brachycera, Cyclorrhapha). Our analysis revealed that the diversification of FCs in these species shows characteristics of both the Rhagio and Drosophila types. First, a distinct cluster of FCs, consisting of polar cells and border-like cells, differentiates at the posterior pole of the ovarian follicle. This feature is unique to the Rhagio type and has only been reported in species representing the Orthorrhapha group. Second, morphological criteria have identified a significantly smaller number of subpopulations of FCs than in Drosophila. Furthermore, while the general pattern of FC migration is similar to that of Drosophila, the distinctive migration of the anterior-dorsal FCs is absent. In the studied tephritids, the migration of the anterior polar cell/border cell cluster towards the anterior pole of the oocyte is followed by the posterior migration of the main body cuboidal FCs to cover the expanding oocyte. Finally, during the onset of vitellogenesis, a distinct subset of FCs migrates towards the centre of the ovarian follicle to cover the oocyte's anterior pole. Our study also highlights specific actions of some FCs that accompany the migration process, which has not been previously documented in cyclorrhaphans. These results support the hypothesis that the posterior and centripetal migrations of morphologically unique FC subsets arose in the common ancestor of Cyclorrhapha. These events appear to have occurred fairly recently in the evolutionary timeline of Diptera., (© 2024 Anatomical Society.)

Moose ( Alces alces ) in boreal habitats feed and rest where they are exposed to Dipteran flies and the parasites they carry. We collected 31,905 flies during the summer from 12 habituated moose on the Kenai Peninsula, Alaska. Moose flies, Haematobosca alcis (Snow), Diptera: Muscidae-a species that completes its entire life cycle on or around moose-accounted for 91% of flies collected; the reminder of the flies collected included mosquitoes (Culicidae), black flies (Simuliidae), and deer flies (Tabanidae). Flies impose physiological costs for moose, e.g., vectors for parasites such as Legworm ( Onchocerca spp.) which causes sores on the hind legs of moose. We found that the number of sores present on the hind legs of moose is positively correlated with body fat, which suggests a correlation between gains of energy and damage from flies. We also found that the number of sores is negatively correlated with serum albumin, which is indicative of an inflammatory response and body protein being used to repair injuries from flies and parasites. The number or type of flies present on a Moose were not correlated with the concentration of corticosteroids in saliva or feces. Flies do not elicit a stress response in moose even though the costs of repairing wounds and resisting infections of those wounds likely reduce gains of protein from summer foraging. Moose can tolerate the injuries from biting flies with regular gains from summer foraging but exposure to insect-borne parasites poses a risk to reproduction and survival., Competing Interests: None declared., (© The Author(s) 2024. Published by Oxford University Press on behalf of the American Society of Mammalogists.)

Moose ( Alces alces ) in the boreal forest habitats of Alaska are unlike other northern ungulates because they tolerate high densities of flies (Diptera) even though flies cause wounds and infections during the warm summer months. Moose move to find food and to find relief from overheating (hyperthermia) but do they avoid flies? We used GPS collars to measure the rate of movement (m⋅h -1 ) and the time spent (min⋅day -1 ) by enclosed moose in four habitats: wetlands, black spruce, early seral boreal forest, and late seral boreal forest. Fly traps were used in each habitat to quantify spatio-temporal abundance. Average daily air temperatures increased into July when peak biomass of forage for moose was greatest in early seral boreal forest habitats (424.46 vs. 25.15 kg⋅ha -1 on average in the other habitats). Average daily air temperatures were 1.7°C cooler in black spruce than other habitats, but fly abundance was greatest in black spruce (approximately 4-fold greater on average than the other habitats). Moose increased their movement rate with counts of biting flies (mosquitoes, black flies, horse and deer flies), but not non-biting flies (coprophagous flies). However, as air temperature increased (above 14.7°C) moose spent more time in fly-abundant black spruce, than early seral boreal forest, showing great tolerance for mosquitoes. Warm summer temperatures appear to cause moose to trade-off foraging in fly-sparse habitats for resting and dissipating heat in shady, wet habitats with abundant flies that adversely affect the fitness of moose., Competing Interests: The authors declare that they have no conflict of interest., (© 2024 The Author(s). Ecology and Evolution published by John Wiley & Sons Ltd. This article has been contributed to by U.S. Government employees and their work is in the public domain in the USA.)

Evidence for parental environments profoundly influencing the physiology, biology, and neurobiology of future generations has been accumulating in the literature. Recent efforts to understand this phenomenon and its underlying mechanisms have sought to use species like rodents and insects to model multi-generational legacies of parental experiences like stress and nutritional exposures. From these studies, we have come to appreciate that parental exposure to salient environmental experiences impacts the cadence of brain development, hormonal responses to stress, and the expression of genes that govern cellular responses to stress in offspring. Recent studies using chemosensory exposure have emerged as a powerful tool to shed new light on how future generations come to be influenced by environments to which parents are exposed. With a specific focus on studies that have leveraged such use of salient chemosensory experiences, this review synthesizes our current understanding of the concept, causes, and consequences of the inheritance of chemosensory legacies by future generations and how this field of inquiry informs the larger picture of how parental experiences can influence offspring biology., (© The Author(s) 2024. Published by Oxford University Press.)

As research on Wolbachia male-killers of dipterans increases over time, several standard methods have emerged for the validation and characterization of these Wolbachia strains. Here, we describe typical steps for maintenance and evaluation of male-killers in the laboratory. Drosophila that host Wolbachia male-killers include both mushroom-feeders and flies reared on standard media, which require different maintenance protocols. In addition, male killing in flies typically occurs during embryonic development, and as such, many experiments require embryo tissue samples. We provide step-by-step instructions for standard methods of isolation, rearing, confirmation of Wolbachia male-killing as the basis of a sex ratio bias, and collection of embryos from these species. Thus, anyone intending to study a male-killer in the lab will have the complete set of protocols necessary to collect and rear them, validate them, and collect embryonic tissue for further study., (© 2024. The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature.)

In Drosophila melanogaster , several Gal4 drivers are used to direct gene/RNAi expression to different dopaminergic neuronal clusters. We previously developed a fly model of Parkinson's disease, in which dopaminergic neurons had elevated cytosolic Ca 2+ due to the expression of a Plasma Membrane Ca 2+ ATPase (PMCA) RNAi under the thyroxine hydroxylase (TH)-Gal4 driver. Surprisingly, TH-Gal4>PMCA RNAi flies died earlier compared to controls and showed swelling in the abdominal area. Flies expressing the PMCA RNAi under other TH drivers also showed such swelling and shorter lifespan. Considering that TH-Gal4 is also expressed in the gut, we proposed to suppress the expression specifically in the nervous system, while maintaining the activation in the gut. Therefore, we expressed Gal80 under the direction of the panneuronal synaptobrevin (nSyb) promoter in the context of TH-Gal4. nSyb-Gal80; TH-Gal4>PMCA RNAi flies showed the same reduction of survival as TH-Gal4>PMCA RNAi flies, meaning that the phenotype of abdomen swelling and reduced survival could be due to the expression of the PMCA RNAi in the gut. In perimortem stages TH-Gal4>PMCA RNAi guts had alteration in the proventriculi and crops. The proventriculi appeared to lose cells and collapse on itself, and the crop increased its size several times with the appearance of cellular accumulations at its entrance. No altered expression or phenotype was observed in flies expressing PMCA RNAi in the dopaminergic PAM cluster (PAM-Gal4>PMCA RNAi ). In this work we show the importance of checking the global expression of each promoter and the relevance of the inhibition of PMCA expression in the gut.

Cochliomyia hominivorax, from the family Calliphoridae, is responsible for causing most myiases in domesticated animals in Brazil. As alternatives to conventional treatments, essential oils (EOs) could possibly be used as natural control of ectoparasites. The aim of this study was to evaluate the in vitro activity of EOs from Salvia sclarea L., Rosmarinus officinalis L., Lavandula hybrida Reverchon, Citrus bergamia Risso, Citrus paradisi L., Juniperus virginiana L., Copaifera reticulata Ducke, Illicium verum Hook f., Pelargonium roseum, Cymbopogon winterianus Jowitt, Cymbopogon flexuosus Steud, Eugenia caryophyllus Spreng, Cinnamomum cassia Presl, Thymus vulgaris L. and Origanum vulgare L. against third-instar larvae of C. hominivorax. To evaluate the mortality rate, filter paper tests were performed at different concentrations and verified after 24 h and 48 h. Essential oils of T. vulgaris, O. vulgare and I. verum induced 100% mortality in the screening test, and their main active components (thymol, carvacrol and anethol, respectively) were also tested alone. In this new test, lethal concentrations (LC 50 ) after 24 h and 48 h, respectively, were 407.1 and 314.2 µg.cm - ² for T. vulgaris, 540.9 and 253.8 µg.cm - ² for O. vulgare, 314.2 µg.cm - ² (after 48 h) for I. verum, 255.6 and 102.3 µg.cm - ² for thymol, 970.5 and 931.1 µg.cm -2 for carvacrol and 559.4 µg.cm -2 (after 48 h) for anethol. These results showed the potential of these EOs and their main components for development of ectoparasiticides for veterinary use, especially T. vulgaris EO and thymol., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 Elsevier B.V. All rights reserved.)

Lameness in sheep is one of the most serious issues on farms in the UK and worldwide, affecting over 90% of all UK sheep flocks. Despite its severity and prevalence, there are knowledge gaps regarding transmission routes of bacterial pathogens associated with infectious lameness in sheep. As larvae of Lucilia sericata are commonly found on foot lesions on lame sheep, it was hypothesised that the flies or their larvae could harbour lameness associated bacteria. This study examined the gut contents of larvae obtained from the foot lesions of lame sheep and compared them to control larvae collected from infested cat food on the same farm. Of particular interest, were the presence of three different bacterial genera associated with lameness; Fusobacterium necrophorum, Dichelobacter nodosus and Treponema spp., for which viability was also investigated. Larvae were cultured In vitro and some allowed to metamorphose into flies before specific PCR assays were carried out on the gut contents. Results showed a significant association between the bacteria on the feet of the sheep and those within the larvae. Although the gut contents of all larvae found on sheep feet contained one or more of the lameness bacteria, none of the bacteria were recovered from the adult flies, suggesting a level of gut remodelling during metamorphosis. Interestingly, Treponema spp. and Fusobacterium spp. were viable when isolated from gut contents of larvae. Maintenance of infection from larvae to fly did not occur. However, it still remains important to control both disease and insect populations of farms to maintain animal welfare., Competing Interests: Declaration of Competing Interest None., (Copyright © 2023 The Authors. Published by Elsevier Ltd.. All rights reserved.)

Background: Histomonas meleagridis can infect chickens and turkeys. It uses the eggs of the cecal worm Heterakis gallinarum as a vector and reservoir. Litter beetles (Alphitobius diaperinus) and other arthropod species have been implicated as potential vectors, but little information about other arthropod species as potential vectors is known., Methods: Four broiler breeder pullet farms were sampled every 4 months. On each farm, three types of traps were set inside and outside two houses. Trapped arthropod specimens were morphologically identified at order level and grouped into families/types when possible. Selected specimens from abundant types found both inside and outside barns were screened for H. meleagridis and H. gallinarum by qPCR., Results: A total of 4743 arthropod specimens were trapped. The three most frequently encountered orders were Diptera (38%), Coleoptera (17%), and Hymenoptera (7%). Three hundred seventeen discrete types were differentiated. More arthropods were trapped outside than inside. Alpha diversity was greater outside than inside but not significantly influenced by season. The composition of the arthropod populations, including the insectome, varied significantly between trap location and seasons. Up to 50% of litter beetles tested positive for H. meleagridis DNA 4 months after an observed histomonosis outbreak. Sporadically litter beetles were positive for H. gallinarum DNA. Thirteen further arthropod types were tested, and specimens of four Dipteran families tested positive for either one or both parasites., Conclusions: This study describes the insectome in and around broiler breeder pullet farms and identifies new potential vectors of H. meleagridis through qPCR. The results show a limited but present potential of arthropods, especially flies, to transmit histomonosis between farms., (© 2023. The Author(s).)

The European rabbit (Oryctolagus cuniculus) populations of the Iberian Peninsula have been severely affected by the emergence of the rabbit haemorrhagic disease virus (RHDV) Lagovirus europaeus/GI.2 (RHDV2/b). Bushflies and blowflies (Muscidae and Calliphoridae families, respectively) are important RHDV vectors in Oceania, but their epidemiological role is unknown in the native range of the European rabbit. In this study, scavenging flies were collected between June 2018 and February 2019 in baited traps at one site in southern Portugal, alongside a longitudinal capture-mark-recapture study of a wild European rabbit population, aiming to provide evidence of mechanical transmission of GI.2 by flies. Fly abundance, particularly from Calliphoridae and Muscidae families, peaked in October 2018 and in February 2019. By employing molecular tools, we were able to detect the presence of GI.2 in flies belonging to the families Calliphoridae, Muscidae, Fanniidae and Drosophilidae. The positive samples were detected during an RHD outbreak and absent in samples collected when no evidence of viral circulation in the local rabbit population was found. We were able to sequence a short viral genomic fragment, confirming its identity as RHDV GI.2. The results suggest that scavenging flies may act as mechanical vectors of GI.2 in the native range of the southwestern Iberian subspecies O. cuniculus algirus. Future studies should better assess their potential in the epidemiology of RHD and as a tool for monitoring viral circulation in the field., (© 2023. The Author(s).)

Linezolid is an antibiotic of last resort for the treatment of infections caused by Gram-positive bacteria, including vancomycin-resistant enterococci. Enterococcus faecalis , a member of enterococci, is a significant pathogen in nosocomial infections. E. faecalis resistance to linezolid is frequently related to the presence of optrA, which is often co-carried with fex , phenicol exporter genes, and erm genes encoding macrolide resistance. Therefore, the common use of antibiotics in veterinary might promote the occurrence of optrA in livestock settings. This is a cross-sectional study aiming to investigate the prevalence of optrA positive E. faecalis (OPEfs) in 6 reservoirs in farms in Ha Nam province, Vietnam, and its associated factors and to explore genetic relationships of OPEfs isolates. Among 639 collected samples, the prevalence of OPEfs was highest in flies, 46.8% (51/109), followed by chickens 37.3% (72/193), dogs 33.3% (17/51), humans 18.7% (26/139), wastewater 16.4% (11/67) and pigs 11.3%, (14/80). The total feeding area and total livestock unit of the farm were associated with the presence of OPEfs in chickens, flies, and wastewater. Among 186 OPEfs strains, 86% were resistant to linezolid. The presence of optrA was also related to the resistant phenotype against linezolid and levofloxacin of E. faecalis isolates. Close genotypic relationships identified by Pulsed Field Gel Electrophoresis between OPEfs isolates recovered from flies and other reservoirs including chickens, pigs, dogs, and wastewater suggested the role of flies in the transmission of antibiotic-resistant pathogens. These results provided warnings of linezolid resistance although it is not used in livestock.

The concept of feedback loops between changes in chemical quality of decomposing organic residues and changes in faunal communities was employed in studying how such feedback loops, representing distinct ecological successional stages, determine decomposition dynamics in soils. A 52-week litterbag decomposition study was superimposed onto an 18-year long term field experiment. Four types of organic residues contrasting in chemical quality (i.e., nitrogen (N), lignin, polyphenols, cellulose) were incorporated into soil annually to assess decomposition and associated meso- and macrofauna communities. In the first 4 weeks after residue incorporation (loop #1), the abundances (densities) of both mesofauna and macrofauna were positively influenced by labile cellulose and N. The mesofauna Collembola and Acari contributed 70-100% and 0-30% to the decomposition, respectively, while the macrofauna beetles and flies contributed 20-90% and 10-66%, respectively. The abundances were highest under groundnut (high N, low lignin) ([1.35 and 0.85 individual number (g dry litter) -1 ] for mesofauna and macrofauna, respectively). The presence of macrofauna at week 2 led to a mass loss (R 2  = 0.67**), indicating that macrofauna preceded mesofauna in degrading residue. In week 8 (transition of loop #2 to #3), only macrofauna (beetles dominated contributing 65%) played an important role in lignin decomposition (R 2  = 0.56**), resulting in a mass loss (R 2  = 0.52**). In week 52 (loop #4) macrofauna, ants (Formicidae) replaced beetles as the dominant decomposers showing a feedback reaction to availability of protected cellulose. The Formicidans contributed 94% to the decomposition and influenced losses of mass (R 2  = 0.36*) and N (R 2  = 0.78***). The feedback loop concept provides a more comprehensive "two-sided" view into decomposition, as regulated simultaneously by two factors, than earlier "one-sided" approaches to soil fauna-mediated decomposition., Competing Interests: The authors declare no competing interests., (© 2023 The Authors.)

Poultry farming is an important activity in animal protein production worldwide, either by laying hen farming or broilers. Over the last decades, the change in production systems with confinement of large numbers of hens has increased productivity and reduced costs; however, it has also increased sanitary issues. In this setting, arthropods that are adapted to poultry houses have gained great importance. They cause direct damage to hens, either by blood spoliation caused by ectoparasites or lesions in the digestive tract (e.g., lesser mealworm) or by indirect damages, by transmitting pathogens or by affecting egg quality, when they attain pest status. Synthetic chemical products comprise the most frequently used control strategy against these pests, with relative efficacy and many side effects. In Europe, some countries also adopt alternative prevention or control measures. In Brazil, however, although there are some groups of researchers that work on developing alternative control, its use is virtually zero. The present review shows a critical overview of this context in Brazil, based on the alternatives that have already been studied and made available, but have not been implemented, yet, and the potential stumbling blocks created by the very poultry market against these advances., (© 2022. Sociedade Entomológica do Brasil.)

Campylobacter is a leading foodborne pathogen, and poultry are a major vehicle for infection. Houseflies play important roles in colonization of broiler flocks with Campylobacter but comparable information for turkey farms is limited. Here, we investigated houseflies as potential vectors for Campylobacter in 28 commercial turkey flocks. We characterized species, genotypes, and the antimicrobial resistance (AMR) profiles of Campylobacter from turkey feces and houseflies in the same turkey house. Of the 28 flocks, 25 yielded Campylobacter from turkey droppings and houseflies, with an average of 6.25 and 3.11 Campylobacter log CFU/g feces and log CFU/fly, respectively. Three flocks were negative for Campylobacter both in turkey feces and in houseflies. Both C. coli and C. jejuni were detected in turkey feces and houseflies, with C. coli more likely to be recovered from houseflies than feces. Determination of Campylobacter species, genotypes, and AMR profiles revealed up to six different strains in houseflies from a single house, including multidrug-resistant strains. For the predominant strain types, presence in houseflies was predictive of presence in feces, and vice versa. These findings suggest that houseflies may serve as vehicles for dissemination of Campylobacter , including multidrug-resistant strains, within a turkey house, and potentially between different turkey houses and farms in the same region.

Synanthropic filth flies transport enteric pathogens from feces to food, which upon consumption poses an infection risk. We evaluated the effect of an onsite sanitation intervention─including fly control measures─in Maputo, Mozambique, on the risk of infection from consuming fly-contaminated food. After enumerating flies at intervention and control sites, we cultured fecal indicator bacteria, quantified gene copies for 22 enteric pathogens via reverse transcription quantitative polymerase chain reaction (RT-qPCR), and developed quantitative microbial risk assessment (QMRA) models to estimate annual risks of infection attributable to fly-contaminated foods. We found that the intervention reduced fly counts at latrine entrances by 69% (aRR = 0.31, [0.13, 0.75]) but not at food preparation areas (aRR = 0.92, [0.33, 2.6]). Half of (23/46) of individual flies were positive for culturable Escherichia coli , and we detected ≥1 pathogen gene from 45% (79/176) of flies, including enteropathogenic E. coli (37/176), adenovirus (25/176), Giardia spp. (13/176), and Trichuris trichiura (12/176). We detected ≥1 pathogen gene from half the flies caught in control (54%, 30/56) and intervention compounds (50%, 17/34) at baseline, which decreased 12 months post-intervention to 43% (23/53) at control compounds and 27% (9/33) for intervention compounds. These data indicate flies as a potentially important mechanical vector for enteric pathogen transmission in this setting. The intervention may have reduced the risk of fly-mediated enteric infection for some pathogens, but infrequent detection resulted in wide confidence intervals; we observed no apparent difference in infection risk between groups in a pooled estimate of all pathogens assessed (aRR = 0.84, [0.61, 1.2]). The infection risks posed by flies suggest that the design of sanitation systems and service delivery should include fly control measures to prevent enteric pathogen transmission.

Background: Huntingtin (htt) protein is an essential regulator of nervous system function through its various neuroprotective and pro-survival functions, and loss of wild-type htt function is implicated in the etiology of Huntington's disease. While its pathological role is typically understood as a toxic gain-of-function, some neuronal phenotypes also result from htt loss. Therefore, it is important to understand possible roles for htt in other physiological circumstances., Objective: To elucidate the role of htt in the context of ethanol exposure, we investigated how loss of htt impacts behavioral and physiological responses to ethanol in Drosophila., Methods: We tested flies lacking htt for ethanol sensitivity and tolerance, preference for ethanol using capillary feeder assays, and recovery of mobility after intoxication. Levels of dopamine neurotransmitter and numbers of dopaminergic cells in brains lacking dhtt were also measured., Results: We found that dhtt-null flies are both less sensitive and more tolerant to ethanol exposure in adulthood. Moreover, flies lacking dhtt are more averse to alcohol than controls, and they recover mobility faster following acute ethanol intoxication. We showed that dhtt mediates these effects at least in part through the dopaminergic system, as dhtt is required to maintain normal levels of dopamine in the brain and normal numbers of dopaminergic cells in the adult protocerebrum., Conclusions: Our results demonstrate that htt regulates the physiological response to ethanol and indicate a novel neuroprotective role for htt in the dopaminergic system, raising the possibility that it may be involved more generally in the response to toxic stimuli.

During decomposition, flies interact with the remains to lay eggs and acquire nutrients, and in the process, they bring their microbes with them. While it is known that flies have their own unique core microbiome, it is not known if flies associated with human cadavers have a different core microbiome. Differences in the fly microbiome may influence the types of microbes transmitted from the flies to the cadaver, therefore potentially affecting assembly of the human decomposer microbiome. The first purpose of this study was to characterize the microbiome of flies associated with human cadavers by fly organ and season. This is because fly interactions with cadavers vary by season, and because it is likely that external fly organs [i.e., the labellum and tarsi] make more direct contact and are likely involved in increased mechanical transmission with the cadaver than internal organs such as the oocyte. The second purpose of this study was to determine if the fly microbes contribute to the human decomposer microbiome. To accomplish these aims, 10 human cadavers were placed outdoors across three seasons and allowed to decompose. A total of 40 flies that landed on the cadaver were collected and dissected by the labellum, tarsi, and oocyte. In addition to fly collections, samples from the cadavers were collected using a sterile swab at sites including the cheek of the face, inner cheek, bicep, torso, and anus. Overall, it was shown that flies associated with human cadavers have a similar microbiome to flies from previous studies that were not associated with human cadavers. However, there are differences in the microbiome between seasons and fly parts. We also show evidence that flies act as a microbial source to the human decomposer microbiome, which is important for understanding the ecological mechanisms of human cadaver microbial community assembly., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2022. Published by Elsevier B.V.)

Lumpy skin disease virus (LSDV) is a poxvirus that causes severe systemic disease in cattle and is spread by mechanical arthropod-borne transmission. This study quantified the acquisition and retention of LSDV by four species of Diptera (Stomoxys calcitrans, Aedes aegypti, Culex quinquefasciatus, and Culicoides nubeculosus) from cutaneous lesions, normal skin, and blood from a clinically affected animal. The acquisition and retention of LSDV by Ae. aegypti from an artificial membrane feeding system was also examined. Mathematical models of the data were generated to identify the parameters which influence insect acquisition and retention of LSDV. For all four insect species, the probability of acquiring LSDV was substantially greater when feeding on a lesion compared with feeding on normal skin or blood from a clinically affected animal. After feeding on a skin lesion LSDV was retained on the proboscis for a similar length of time (around 9 days) for all four species and for a shorter time in the rest of the body, ranging from 2.2 to 6.4 days. Acquisition and retention of LSDV by Ae. aegypti after feeding on an artificial membrane feeding system that contained a high titer of LSDV was comparable to feeding on a skin lesion on a clinically affected animal, supporting the use of this laboratory model as a replacement for some animal studies. This work reveals that the cutaneous lesions of LSD provide the high-titer source required for acquisition of the virus by insects, thereby enabling the mechanical vector-borne transmission. IMPORTANCE Lumpy skin disease virus (LSDV) is a high consequence pathogen of cattle that is rapidly expanding its geographical boundaries into new regions such as Europe and Asia. This expansion is promoted by the mechanical transmission of the virus via hematogenous arthropods. This study quantifies the acquisition and retention of LSDV by four species of blood-feeding insects and reveals that the cutaneous lesions of LSD provide the high titer virus source necessary for virus acquisition by the insects. An artificial membrane feeding system containing a high titer of LSDV was shown to be comparable to a skin lesion on a clinically affected animal when used as a virus source. This promotes the use of these laboratory-based systems as replacements for some animal studies. Overall, this work advances our understanding of the mechanical vector-borne transmission of LSDV and provides evidence to support the design of more effective disease control programmes.

Multi-drug resistant pathogens are a global problem. Flies are a potential vector of multi-drug resistant pathogens, which can be particularly dangerous in the hospital environment. This study aimed to evaluate flies as vectors of alert pathogens. The research material consisted of 100 flies ( Musca domestica (46.0%), Lucilia sericata (28.0%), and Calliphora vicina (26.0%)) collected at the University Hospital No. 1 dr. A. Jurasz in Bydgoszcz (Poland) in 2018-2019 (summer months). The presence of bacteria of the genera: Enterococcus, Staphylococcus, Escherichia, Leclercia, Citrobacter, Hafnia, Providencia, Proteus, Enterobacter, Klebsiella, Raoultella, Morganella, Moellerella, Bordetella, Pantoea, Serratia, Plesiomonas, Wohlfahrimonas , and Lelliottia was confirmed. The most frequently isolated species included: Enterococcus faecalis (n = 64), Escherichia coli (n = 43) and Moellerella wisconsensis (n = 24). The infection rate and antibiotic resistance of bacteria were assessed. One strain of Proteus mirabilis (isolated from Calliphora vicina) produced ESBLs (extended-spectrum beta-lactamases). The infection rate was 0.38%, 0.26%, and 0.20% for Musca domestica, Lucilia sericata , and Calliphora vicina , respectively. The flies from a hospital area were not a vector of alert pathogens. Monitoring flies as potential vectors of pathogens is an important aspect of public health, especially for hospitalized patients.

Insect life cycles are short-term and therefore sensitive to immediate changes triggered by climate, vegetation structure, or land use management; hence, the insect populations shape the communities and functional relationships in tropical forests. In this study, we hypothesized that seasonal variations of the dipteran families respond in different ways to changes of weather conditions, thereby affecting their population dynamics. In a one-hectare plot, we surveyed the fly community inhabiting the understory of a Neotropical rainforest. Over a yearly cycle, we used three Malaise traps operated continuously for 365 days and recorded a total of 68,465 fly specimens belonging to 48 families of Diptera, 15 of which were most abundant, accounting for 99.2% of all sampled individuals. The results of the trapping frequency indices (TFIs) exhibited significant population fluctuations in 12 of the 15 most abundant families, which were particularly correlated with temperature or precipitation. Based on such variations, we identified four seasonal patterns as follows: (i) Spring-Autumn bimodal pattern (Cecidomyiidae, Sciaridae, Phoridae, Stratiomyidae); (ii) Spring pattern (Mycetophilidae, Dolichopodidae, Ceratopogonidae); (iii) Autumn pattern (Chironomidae, Psychodidae); (iv) Winter pattern (Empididae, Tipulidae, Ditomyiidae). From a functional perspective, we found the prevalence of families with saprophagous larvae, in addition to phytophagous, fungivores, and predacious. Our results suggest a key role played by the Diptera community on structuring the functional clusters, both in terms of taxonomic composition and on seasonal shifts of abundance, thus influencing the dynamic processes of nutrient cycling in the understory., (© 2022. Sociedade Entomológica do Brasil.)

The potential of insects for forensic investigations has been known for more than 700 years. However, arthropods such as mites could also play a role in these investigations. The information obtained from insects, together with their phoretic mites, is of special interest in terms of estimating the time and geographical location of death. This paper presents the first interaction network between phoretic mites and their host insects in Navarra. It also reports the first time that an interaction network was applied to animal remains of forensic relevance. The data reveal the degrees of specificity of the interactions established, the biological and ecological characteristics of the mites at the time of association, and factors that played important roles in the mites' dispersion. Fauna was collected using 657 traps baited with 20 g of pig carrion over a year. Only 0.6% of insects collected carried phoretic mites. The network comprised 312 insects (275 beetles, 37 flies) and 1533 mites and was analyzed using various packages of the R programming language. We contribute new host insect records for 15 mites, 3 new records of insects as hosts, 5 new mite records for the Iberian Peninsula, and 2 new mites records and 8 new insect records for Navarra.

Liver Kinase B1 (LKB1) is known as a master kinase for 14 kinases related to the adenosine monophosphate-activated protein kinase. Two of them salt inducible kinase 3 and adenosine monophosphate-activated protein kinase α have previously been implicated in sleep regulation. We generated loss-of-function mutants for Lkb1 in both Drosophila and mice. Sleep, but not circadian rhythms, was reduced in Lkb1-mutant flies and in flies with neuronal deletion of Lkb1. Genetic interactions between Lkb1 and threonine to alanine mutation at residue 184 of adenosine monophosphate-activated protein kinase in Drosophila sleep or those between Lkb1 and Threonine to Glutamic Acid mutation at residue 196 of salt inducible kinase 3 in Drosophila viability have been observed. Sleep was reduced in mice after virally mediated reduction of Lkb1 in the brain. Electroencephalography analysis showed that nonrapid eye movement sleep and sleep need were both reduced in Lkb1-mutant mice. These results indicate that liver kinase B1 plays a physiological role in sleep regulation conserved from flies to mice., (© The Author(s) 2022. Published by Oxford University Press on behalf of Genetics Society of America. All rights reserved. For permissions, please email: journals.permissions@oup.com.)

The unique biology of flies and their omnipresence in the environment of people and animals makes them ideal candidates to be important vectors of antimicrobial resistance genes. Consequently, there has been increasing research on the bacteria and antimicrobial resistance genes that are carried by flies and their role in the spread of resistance. In this review, we describe the current knowledge on the transmission of bacterial pathogens and antimicrobial resistance genes by flies, and the roles flies might play in the maintenance, transmission, and surveillance of antimicrobial resistance.

Camellia oleifera Abel. is an important woody oil plant, and its pollination success is essential for oil production. We conducted this study to select the best pollinator candidates for C. oleifera using principal component analysis and multi-attribute decision-making. Field observations of the flower-visiting characteristics of candidate pollinators were conducted at three sites. The insect species that visited flowers did not considerably differ between regions or time periods. However, the proportion of each species recorded did vary. We recorded eleven main candidates from two orders and six families at the three sites. The pollen amount carried by Apis mellifera was significantly higher than that of other insects. However, the visit frequency and body length of Apis mellifera were smaller than those of Vespa velutina . Statistical analysis showed that A. mellifera is the best candidate pollinator; Eristalis cerealis is a good candidate pollinator; Phytomia zonata , A. cerana, and V. velutina were ordinary candidate pollinators; and four fly species, Episyrphus balteatus were classified as inefficient candidate pollinators. Our study shows that flies and hoverflies play an important role in the pollination system. Given the global decline in bee populations, the role of flies should also be considered in Eristalinus arvorum were classified as inefficient candidate pollinators. Our study shows that flies and hoverflies play an important role in the pollination system. Given the global decline in bee populations, the role of flies should also be considered in C. oleifera seed production.

Over the last decade, the increasing interest in long non-coding RNAs (lncRNAs) has led to the discovery of these transcripts in multiple organisms. LncRNAs tend to be specifically, and often lowly, expressed in certain tissues, cell types and biological contexts. Although lncRNAs participate in the regulation of a wide variety of biological processes, including development and disease, most of their functions and mechanisms of action remain unknown. Poor conservation of the DNA sequences encoding for these transcripts makes the identification of lncRNAs orthologues among different species very challenging, especially between evolutionarily distant species such as flies and humans or mice. However, the functions of lncRNAs are unexpectedly preserved among different species supporting the idea that conservation occurs beyond DNA sequences and reinforcing the potential of characterising lncRNAs in animal models. In this review, we describe the features and roles of lncRNAs in the fruit fly Drosophila melanogaster, focusing on genomic and functional comparisons with human and mouse lncRNAs. We also discuss the current state of advances and limitations in the study of lncRNA conservation and future perspectives., (© 2021. The Author(s).)

Antimicrobial resistance (AMR) in bacteria in the livestock is a growing problem, partly due to inappropriate use of antimicrobial drugs. Antimicrobial use (AMU) occurs in Swedish dairy farming but is restricted to the treatment of sick animals based on prescription by a veterinary practitioner. Despite these strict rules, calves shedding antimicrobial resistant Enterobacteriaceae have been recorded both in dairy farms and in slaughterhouses. Yet, not much is known how these bacteria disseminate into the local environment around dairy farms. In this study, we collected samples from four animal sources (fecal samples from calves, birds and rodents, and whole flies) and two environmental sources (cow manure drains and manure pits). From the samples, Escherichia coli was isolated and antimicrobial susceptibility testing performed. A subset of isolates was whole genome sequenced to evaluate relatedness between sources and genomic determinants such as antimicrobial resistance genes (ARGs) and the presence of plasmids were assessed. We detected both ARGs, mobile genetic elements and low rates of AMR. In particular, we observed four potential instances of bacterial clonal sharing in two different animal sources. This demonstrates resistant E. coli dissemination potential within the dairy farm, between calves and scavenger animals (rodents and flies). AMR dissemination and the zoonotic AMR risk is generally low in countries with low and restricted AMU. However, we show that interspecies dissemination does occur, and in countries that have little to no AMU restrictions this risk could be under-estimated., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2022 Hickman, Agarwal, Sjöström, Emanuelson, Fall, Sternberg-Lewerin and Järhult.)

Foodborne pathogens have become a major concern not only due to the diseases they cause, but also because of the rise of antibiotic resistant strains in human and animals. The purposes of this study were to determine the occurrence of  Campylobacter jejuni and Escherichia coli and their antibiotic resistance profiles in wild birds, chickens, humans, and the environment in Malay villages in Malaysia. Three Malay villages in Kota Setar, Kedah were chosen. Three hundred nine (309) samples were collected in this study including wild birds (38), chickens (71), humans (47), and the environment (153). Subsequently, the C. jejuni and E   coli isolates were tested against antibiotics using the disc diffusion method. Campylobacter jejuni was found positive in 17 (37.8%) flies and 8 (11.3%) chickens. Also, E. coli was found positive in 89.4% of human, 47.4% of bird, 44 62% of chicken and in 71.2% of the environmental samples. Ten antibiotics were used to determine the susceptibility of the isolates. Eighty four percent (84%) of  C. jejuni and 100% of  E. coli isolates were found to show resistance towards at least one antibiotic. The isolates showed high resistance to cefpodoxime and tetracycline., Competing Interests: The authors declare no conflict of interest., (Copyright: © 2022 Ibrahim Mohamed et al.)

The abdominal and pregenital segments and the genitalia were studied in males of Fannia subpellucens (Zetterstedt, 1845), Fannia canicularis (Linnaeus, 1761) and Fannia incisurata (Zetterstedt, 1838). In comparison with the remaining members of the muscoid grade, in addition to the symmetry of the pregenital segments, significant reductions of the sclerites and musculature of the male terminalia have been observed in Fanniidae. The muscular structure of pregenital segments confirms that the fused pregenital ring is syntergosternite VI + VII + VIII. Symmetry and fusion, as well as the lower number of the sclerites and muscles of the pregenital segments and male genitalia of the Fanniidae, can be considered apomorphic character states. The presence of the lateral bacilliform sclerite, as well as the presence and position of the epandrial muscles M 26, three pairs of muscles M 19 and paired muscles M 18, can be considered as a plesiomorphic character state of the Fanniidae. The structure of the sclerites and muscles of the male abdominal segments and terminalia place the Fanniidae at the base of the muscoid grade and Oestroidea, as has been confirmed by recent molecular studies.

Plant-pollinator interactions provide a natural experiment in signal evolution. Flowers are known to have evolved colour signals that maximise their ease of detection by the visual systems of important pollinators such as bees. Whilst most angiosperms are bee pollinated, our understanding on how the second largest group of pollinating insects, flies, may influence flower colour evolution is limited to the use of categorical models of colour discrimination that do not reflect the small colour differences commonly observed between and within flower species. Here we show by comparing flower signals that occur in different environments including total absence of bees, a mixture of bee and fly pollination within one plant family (Orchidaceae) from a single community, and typical flowers from a broad taxonomic sampling of the same geographic region, that perceptually different colours, empirically measured, do evolve in response to different types of insect pollinators. We show evidence of both convergence among fly-pollinated floral colours but also of divergence and displacement of colour signals in the absence of bee pollinators. Our findings give an insight into how both ecological and agricultural systems may be affected by changes in pollinator distributions around the world., (© 2021 The Authors. New Phytologist © 2021 New Phytologist Foundation.)

Background: Due to the confirmation of the presence of the novel coronavirus in the feces and municipal sewerage system, and the feeding of domestic insects from fecal matter, as well as the ability of these insects to mechanically transmit microbes from the sewerage system. This study was aimed at molecular evaluation of the novel coronavirus infection isolated on cockroaches and flies collected from Kamkar-Arabnia Hospital in Qom City, Iran., Methods: Totally, 18 samples; (12 samples cockroaches and 6 flies) from the external surface of cockroaches and houseflies as well as their digestive system were prepared. After designed and synthetized exogenous heterologous internal control, the RNA was extracted to investigate the contamination of these samples with the novel coronavirus. To detect the virus, the E and RdRp genes were identified., Results: Investigation of coronavirus E gene using the multiplex one-step qPCR technique on the collected samples showed an amplification plot in CT= 35.70 related to the internal surfaces of cockroaches collected from the treatment and sick room of the hospital. Also, the design of internal control to ensure the accuracy of the extraction process was successful., Conclusion: According to the findings of the present study regarding detecting the presence of the coronavirus infection in the digestive system of domestic insects such as American cockroaches and considering their ability to mechanically transmit viruses, it is recommended to control the domestic insects that are in close contact with humans in crowded places such as hospitals and health centers during the Covid-19 pandemic., Competing Interests: Conflict of interest statement Authors declare that there is no conflict of interest., (Copyright © 2021 The Authors. Published by Tehran University of Medical Sciences.)

The current study aimed at molecular identification and comparing the diversity of arthropods communities between pig and sheep carcasses during the cold and warm season in KwaZulu-Natal province of South Africa. Adult arthropods found on and around the carcasses were collected using either fly traps or forceps. Molecular analyses confirmed the identification of twelve arthropod species collected from both sheep and pig carcasses during the cold season. Results showed that 11 of 12 arthropod species were common in both sheep and pig carcasses, with exception to Onthophagus vacca (Coleoptera: Scarabaeidae) (Linnaeus, 1767) and Atherigona soccata (Diptera: Muscidae) (Rondani, 1871) species which were unique to sheep and pig carcasses respectively. However, during the warm season, the sheep carcass attracted more arthropod ( n = 13) species as compared to the pig carcass. The difference in the obtained arthropod was due to the presence of O. vacca which was also unique to the sheep carcass during this season. Furthermore, there was an addition of a beetle species Hycleus lunatus (Coleoptera: Meloidae) (Pallas, 1782), which was collected from both sheep and pig carcasses but unique to the warm season. The pig carcass attracted more dipteran flies during both warm ( n = 1,519) and cold season ( n = 779) as compared to sheep carcass during the warm ( n = 511) and cold season ( n = 229). In contrast, coleopterans were more abundant on the sheep carcass during the warm season ( n = 391) and cold season ( n = 135) as compared to the pig carcass in both warm season ( n = 261) and cold season ( n = 114). In overall, more flies and beetles were collected on both sheep and pig carcasses during the warm season, and this further highlight that temperature influenced the observed difference in the abundance of collected arthropod between seasons., Competing Interests: The authors declare that they have no competing interests., (© 2021 Tembe et al.)

The structural patterns comprising bimodal pollination networks can help characterize plant-pollinator systems and the interactions that influence species distribution and diversity over time and space. We compare network organization of three plant-pollinator communities along the altitudinal gradient of the San Francisco Peaks in northern Arizona. We found that pollination networks become more nested, as well as exhibit lower overall network specialization, with increasing elevation. Greater weight of generalist pollinators at higher elevations of the San Francisco Peaks may result in plant-pollinator communities less vulnerable to future species loss due to changing climate or shifts in species distribution. We uncover the critical, more generalized pollinator species likely responsible for higher nestedness and stability at the higher elevation environment. The generalist species most important for network stability may be of the greatest interest for conservation efforts; preservation of the most important links in plant-pollinator networks may help secure the more specialized pollinators and maintain species redundancy in the face of ecological change, such as changing climate.

Diptera is one of the biggest insect orders and displays a large diversity of visual adaptations. Similarly to other animals, the dipteran visual process is mediated by opsin genes. Although the diversity and function of these genes are well studied in key model species, a comprehensive comparative genomic study across the dipteran phylogeny is missing. Here we mined the genomes of 61 dipteran species, reconstructed the evolutionary affinities of 528 opsin genes, and determined the selective pressure acting in different species. We found that opsins underwent several lineage-specific events, including an independent expansion of Long Wave Sensitive opsins in flies and mosquitoes, and numerous family-specific duplications and losses. Both the Drosophila and the Anopheles complement are derived in comparison with the ancestral dipteran state. Molecular evolutionary studies suggest that gene turnover rate, overall mutation rate, and site-specific selective pressure are higher in Anopheles than in Drosophila. Overall, our findings indicate an extremely variable pattern of opsin evolution in dipterans, showcasing how two similarly aged radiations, Anopheles and Drosophila, are characterized by contrasting dynamics in the evolution of this gene family. These results provide a foundation for future studies on the dipteran visual system., (© The Author(s) 2021. Published by Oxford University Press on behalf of the Society for Molecular Biology and Evolution.)

The abandonment of historical land-use forms within forests, such as grazing or coppicing, and atmospheric nitrogen deposition, has led to an increasing overgrowth of forest gaps and canopy closure in forest ecosystems of Central Europe. From 1945 to 2015, 81% of the forest gaps greater than 150 m 2 within the study area transitioned into a closed forest.This study investigated how the overgrowth process affects flower supply, flower visitors, and reproduction of Campanula species. Six native Campanula species with different light requirements were used as phytometers.The forest gaps in the studied area are a feature of the historical European cultural landscape. We compared large gaps caused by human activities, small gaps caused by habitat conditions, and closed forests. In eight blocked replicates, each with the three habitat categories, we recorded the flower cover and number of indigenous flowering species in the immediate surroundings, and, of six Campanula species, flower visitors and seed production.Forest gaps and their size positively affected the number of flowering plant species in the surrounding area, the number of all flower visitor groups, and the number of seeds produced by all six Campanula species. Flower cover in the surrounding area was higher in large gaps, but there was no difference between small gaps and closed forests. Among flower visitors, small bees varied the most between the three habitat categories, and flies varied the least. The effect on the number of seeds produced was particularly strong for three light-demanding Campanula species.The overgrowth of forest gaps negatively affected flower supply, flower-visiting insects, and seed sets of six Campanula species. Forest gaps should be managed to maintain the reproduction of open forest plants and their pollinators., Competing Interests: None declared., (© 2021 The Authors. Ecology and Evolution published by John Wiley & Sons Ltd.)

Ethanol at low doses induces a locomotor stimulant response across a range of phylogenetically diverse species. In rodents, this response is commonly used as an index of ethanol's disinhibitory, anxiolytic, or reinforcing effects, and its expression is regulated by signaling through a number of conserved neurotransmitter systems. In the current experiments, we asked whether ethanol-induced locomotor stimulation in the fruit fly Drosophila melanogaster might be mediated by ionotropic GABA receptors. We measured basal and ethanol-stimulated locomotion in flies expressing RNAi directed against three known subunits of ionotropic GABA receptors, and also examined the effects of picrotoxin feeding on these behaviors. We found that RNAi-mediated knockdown of a subunit of fly ionotropic GABA receptors, RDL, in all neurons resulted in an increased ethanol-induced locomotor stimulant response, while knockdown of two other subunits, LCCH3 and GRD, did not affect the responses. The effect of pan neuronal RDL knockdown was recapitulated with selective RDL knockdown in cholinergic neurons, and increased ethanol-induced locomotor stimulation was also seen by feeding the GABA A antagonist picrotoxin to flies prior to behavioral testing. However, the increase in ethanol-stimulated locomotion in each of these experiments was largely accounted for by decreased baseline activity. Our results indicate that ionotropic GABA receptors might be a conserved mediator of the locomotor stimulant effects of ethanol, but that alternative experimental approaches will be necessary to disentangle effects of GABAergic manipulations on baseline and ethanol-stimulated locomotion in flies., (Copyright © 2021. Published by Elsevier B.V.)

Abstract: Flies are a vector for spreading foodborne pathogens pertinent to fresh produce, such as Shiga toxigenic Escherichia coli and Salmonella; however, most studies focus on concentrated animal feeding operations, which do not reflect low-density animal farming practices that often adjoin fruit and vegetable acreage. In this study, we determined the prevalence of Salmonella in flies collected biweekly on an integrated animal and produce operation over two growing seasons. Eleven of 889 pooled samples tested positive for Salmonella. Flies from the Calliphoridae, Muscidae, Sarcophagidae, and Tachinidae families were associated with Salmonella carriage, but fly family was not a significant factor for isolation of Salmonella (P = 0.303). Fly species were a significant factor (P = 0.026), with five Pentacricia aldrichii pools testing positive for Salmonella. With the exception of single specimen isolation, prevalence ranged from 2.2 to 15.2%. With the exception of the Tachinidae family, these results reflect a strong association of flies that are commonly associated with feces or are pests of animals. Trap location was not significantly associated with isolation of Salmonella-positive flies (P = 0.236). Overall, the population of flies was not as abundant as studies conducted with produce grown close to concentrated animal feeding operations, indicating a reduced risk of transmission; however, similar to these studies, fly families that are commonly isolated from fecal and decaying matter were most frequently associated with Salmonella isolation. Further work is warranted to elucidate the foodborne pathogen transmission rates to produce and subsequent survival over time., (Published 2021 by the International Association for Food Protection.)

Introduction: synanthropic flies are sometimes involved in the transmission of diarrheal diseases as mechanical vectors of pathogenic bacteria. The purpose of this study was to assess the diversity of synanthropic flies and to determine their potential involvement in the transmission of diarrheal diseases in the city of Maroua., Methods: fly catching sessions were carried out per season in 12 wards, in five different sites and in three moments of the day, corresponding to the different daily sunshine hours. Multiple keys for the identification of diptera and of microbiological analyzes in the laboratory were used to estimate the biodiversity and the portage of microorganisms by the synanthropic flies. We carried out an ecological and statistical analyses of collected data., Results: eight species of synanthropic flies belonging to four families were identified in the city of Maroua and the distribution of these species varied according to the seasons, sites and moment of the day (p<0.05). Musca domestica and Chrysomya putoria were the most numerous species detected in the sites where the activities of agri-food processing and livestock were intense, notably Hardé, Pont-vert, Doualaré, Kongola and Makabaye. Escherichia coli was more involved than Salmonella spp.in the transmission of diarrheal diseases and the portage of bacteria by synanthropic flies was abundance-dependent., Conclusion: the diversity of synanthropic flies varies as a function of anthropogenic activities, season of the year and moment of the day. These flies are important potential mechanical vectors of fecal pathogenic bacteria in Maroua., Competing Interests: Les auteurs ne déclarent aucun conflit d´intérêts., (Copyright: Daniel Amani Dawaye et al.)

Manganese (Mn) is an essential element that, in excess, seems to be involved in the development of different neurodegenerative conditions. Gamma-oryzanol (Ory) was previously reported to possess antioxidant and neuroprotective properties. Thus, we conducted this study to test the hypothesis that Ory can also protect flies in an Mn intoxication model. Adult wild-type flies were fed over 10 days with Mn (5 mM) and/or Ory (25 μM). Flies treated with Mn had a decrease in locomotor activity and a higher mortality rate compared to those in controls. Mn-treated flies also had a significant increase in acetylcholinesterase (AChE) activity, in Mn accumulation and in oxidative stress markers. Moreover, flies treated with Mn exhibited a significant decrease in dopamine levels and in tyrosine hydroxylase activity, as well as in mitochondrial and cellular viability. Particularly important, Ory protected against mortality and avoided locomotor and biochemical changes associated with Mn exposure. However, Ory did not prevent the accumulation of Mn. The present results support the notion that Ory effectively attenuates detrimental changes associated with Mn exposure in Drosophila melanogaster, reinforcing its neuroprotective action/potential.

Wing interference patterns (WIPs) are stable structural colours displayed on insect wings which are only visible at specific viewing geometries and against certain backgrounds. These patterns are widespread among flies and wasps, and growing evidence suggests that they may function as species- and sex-specific mating cues in a range of taxa. As such, it is expected that WIPs should differ between species and show clear sexual dimorphisms. However, the true extent to which WIPs vary between species, sexes and individuals is currently unclear, as previous studies have only taken a qualitative approach, without considering how WIPs might be perceived by the insect. Here, we perform the first quantitative analysis of inter- and intra-specific variation in WIPs across seven Australian species of the blowfly genus Chrysomya. Using multispectral digital imaging and a tentative model of blowfly colour vision, we provide quantitative evidence that WIPs are species-specific, highlight that the extent of divergence is greater in males than in females and demonstrate sexual dimorphisms in several species. These data suggest that WIPs have diversified substantially in blowflies as a result of either sexual or ecological selection., (© 2021 European Society for Evolutionary Biology. Journal of Evolutionary Biology © 2021 European Society for Evolutionary Biology.)

Objectives: Flies have been implicated in the dispersal of medically important bacteria including members of the genus Klebsiella between different environmental compartments. The aim of this study was to retrieve and characterize antibiotic-resistant bacteria from flies collected near to hospitals., Methods: Flies were collected in the vicinity of medical facilities and examined for bacteria demonstrating phenotypic resistance to ceftriaxone, followed by determination of phenotypic and genotypic resistance profiles. In addition, whole genome sequencing followed by phylogenetic analysis and resistance genotyping were performed with the multidrug-resistant (MDR) strain Lemef23, identified as Klebsiella quasipneumoniae subsp. similipneumoniae., Results: The strain Lemef23, classified by multiple locus sequence typing as novel ST 3397, harboured numerous resistance genes. The bla NDM was located on a Tn3000 element, a common genetic platform for the carriage of this gene in Brazil. Inference of phylogenetic orthology of strain Lemef23 and other clinical isolates suggested an anthropogenic origin., Conclusions: The findings of this study support the role of flies as vectors of MDR bacteria of clinical importance and provide the first record of bla NDM-1 and bla CTXM-15 in a Brazilian isolate of K. quasipneumoniae subsp. similipneumoniae, demonstrating the value of surveying insects as reservoirs of antibiotic resistance., (Copyright © 2020 The Authors. Published by Elsevier Ltd.. All rights reserved.)

Necrophagous blowflies (Diptera: Calliphoridae) such as Calliphora vicina, a cold-tolerant species, and Lucilia sericata, a warm-adapted species, are pioneer carrion-breeder. Although these two species have different temperature preferences, larvae aggregate actively and often feed simultaneously on carrion. The hypothesis to be tested was that L. sericata benefits from the association with C. vicina at lower temperatures (i.e., 15 °C) and that C. vicina derives greater benefits from this association at higher temperatures (i.e., 28 °C). Therefore, both species were raised at these two constant temperatures from first instars to adults under three different conditions: monospecific low-density, monospecific high-density, and heterospecific high-density. The time until larval migration, surface area of puparia, and survival rates were determined for each condition. Differences between these fitness-related traits were found between species, temperatures, group densities, and species compositions. At 28 °C, C. vicina larvae bred in heterospecific groups migrated significantly earlier and in higher numbers than that under same density conspecific conditions, with a lower mortality rate. At 15 °C, both species benefited from high-density heterospecific associations, expressed by faster development and larger puparia. In conclusion, necrophagous larvae benefited from heterospecific aggregations at suboptimal temperatures by adapting their migration time to that of the faster species. Since temperature changes throughout the day and over the year, the beneficiary of such a collective association also changes. The costs involved and deviations to the temperature-size rule highlight the complexity of the carrion ecosystem., (© 2020 Institute of Zoology, Chinese Academy of Sciences.)

Urbanization is increasing worldwide and is regarded a major threat to biodiversity in forests. As consequences of intensive human use, the vegetation structure of naturally growing urban forests and their amount of deadwood can be reduced. Deadwood is an essential resource for various saproxylic insects and fungi. We assessed the effects of urbanization and forest characteristics on saproxylic insects and fungi. We exposed standardized bundles consisting of each three freshly cut beech and oak branches in 25 forests along a rural-urban gradient in Basel (Switzerland). After an exposure of 8 months, we extracted the saproxylic insects for 10 months using an emergence trap for each bundle. We used drilling chips from each branch to determine fungal operational taxonomic units (OTUs). In all, 193,534 insect individuals emerged from the experimental bundles. Our study showed that the abundance of total saproxylic insects, bark beetles, longhorn beetles, total flies, moths, and ichneumonid wasps decreased with increasing degree of urbanization, but not their species richness. However, the taxonomic composition of all insect groups combined was altered by wood moisture of branches and that of saproxylic beetles was influenced by the degree of urbanization. Unexpectedly, forest size and local forest characteristics had a minor effect on saproxylic insects. ITS (internal transcribed spacer of rDNA) analysis with fungal specific primers revealed a total of 97 fungal OTUs on the bundles. The number of total fungal OTUs decreased with increasing degree of urbanization and was affected by the volume of naturally occurring fine woody debris. The composition of fungal OTUs was altered by the degree of urbanization and pH of the branch wood. As a consequence of the altered compositions of saproxylics, the association between total saproxylic insects and fungi changed along the rural-urban gradient. Our study shows that urbanization can negatively impact saproxylic insects and fungi., Competing Interests: The authors declare no conflict of interest., (© 2021 The Authors. Ecology and Evolution published by John Wiley & Sons Ltd.)

Digital dermatitis is a bacterial infection that causes lesions above the heel bulbs on cattle hooves, and several bacterial species from the genus Treponema are suspected to be causative agents of this polymicrobial condition. Transmission of the bacteria to healthy cows is understudied, particularly with regard to potential insect vectors. Therefore, the objective of this research was to determine if flies captured from a dairy farm known to have digital dermatitis are contaminated with Treponema bacteria. The DNA-based assays did not detect any Treponema phagedenis from stable flies and house flies collected at a dairy experiencing an outbreak of digital dermatitis. Other potential means of bacterial transmission are discussed., (Copyright © 2021 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.)

Ectoparasites and livestock-associated insects are a major concern throughout the world because of their economic and welfare impacts. Effective control is challenging and relies mainly on the use of chemical insecticides and acaricides. Wolbachia, an arthropod and nematode-infecting, maternally-transmitted endosymbiont is currently of widespread interest for use in novel strategies for the control of a range of arthropod-vectored human diseases and plant pests but to date has received only limited consideration for use in the control of diseases of veterinary concern. Here, we review the currently available information on Wolbachia in veterinary ectoparasites and disease vectors, consider the feasibility for use of Wolbachia in the control of livestock pests and diseases and highlight critical issues which need further investigation., (Crown Copyright © 2020. Published by Elsevier B.V. All rights reserved.)

Forensic entomotoxicology integrates toxicology into forensic entomology to estimate minimum postmortem interval (PMI min ) and circumstances of death where toxicants and poisonous substances are the suspected cause of death. Forensic entomotoxicology not only confirms the presence of toxicants in insects feeding off a cadaver but also studies its effect on the bio-morphometry and growth rate of insects. This review article highlights the effects of various toxicants on forensically important species of dipteran flies. It also discusses the parameters that may affect accuracy in estimation of time since death. The bio-physical effects of toxicants (excluding the analytical approach for qualitative detection) would help understand the trends in forensic entomotoxicological research worldwide.