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.

Thelazia lacrymalis in horses from Romania: epidemiology, morphology and phylogenetic analysis.

BACKGROUND: Equine thelaziosis is a neglected vector-borne parasitic disease in modern veterinary medicine, lacking recent reports. It is transmitted by Musca autumnalis, and potentially other Muscidae species, by ingesting the lachrymal secretions of its equine host. The distribution of both Thelazia lacrymalis and its intermediate hosts remains largely unknown throughout Europe, with most studies dating back 20 years. The aim of this study was to assess the presence, prevalence and distribution of T. lacrymalis in horses from Romania. METHODS: The eyes of 273 horses, slaughtered at two abattoirs from the Northwestern and Western regions of Romania, were examined for the presence of T. lacrymalis between March and November 2021. Upon detection, the nematodes were collected and morphologically identified using the keys from literature. Following identification, one specimen from each animal was selected for molecular analysis while the rest underwent detailed morphometric measurements. Mapping and distribution, according to ecoregions, was done using the QGis 3.20 software, while sequences obtained were compared to those available in GenBank through BLAST analysis using the MEGA X software. RESULTS: Of the 273 animals sampled, 12 (4.39%) were positive for Thelazia spp. INFECTION: Eighty-seven nematodes were recovered, all morphologically identified as T. lacrymalis. The intensity of infestation varied between one and 33 nematodes/animal while five animals presented a bilateral infestation and seven a unilateral one. The highest prevalence was encountered in Pannonian ecoregion (12.12%) while the lowest was in the Alpine ecoregion (0%). Seventy-five intact specimens underwent detailed morphometric analysis, of the 18-20 parameters, resulting in notable differences in striation lengths compared to the data available in other reports. BLAST analysis identified a 96.46-98.60% similarity to the only other COI gene sequence available for T. lacrymalis. CONCLUSIONS: The current study represents the first report of T. lacrymalis in horses in Romania. The low prevalence rates are probably linked to the wide use of macrocyclic lactones.

Parabronema skrjabini (Nematoda: Habronematidae) infection and development in the intermediate host-Haematobia irritans (Linnaeus, 1758) in Inner Mongolia, China.

Parabronemosis is a disease that severely threatens camel health, causing huge economic losses to industries involved in camel husbandry. Previous studies have reported that horn flies (Haematobia irritans) act as intermediate hosts of Parabronema skrjabini; however, the infection and developmental processes of P. skrjabini in horn flies remain unclear. In the present study, the infection rates of P. skrjabini were determined in morphologically and molecularly identified horn flies collected from Bactrian camels (Camelus bactrianus) producing regions in Inner Mongolia, China that have high P. skrjabini infection rates. The horn flies were dissected to obtain the nematode larvae at various instar stages. The P. skrjabini found in the different instar stages of horn fly instars were counted and identified to assess the infection and developmental status. Nematode larvae at different developmental stages were obtained from the horn fly instars for further molecular analysis. Sequencing results confirmed that the nematode larvae were P. skrjabini. Furthermore, we found that the mean growth rate of the nematode larva increased as the horn fly instars develops. The results suggested that P. skrjabini infected the horn flies at the larval instar stage, and that the nematode larvae developed simultaneously with the horn fly instars stages. Our findings provide useful information into the elucidation of P. skrjabini infection and life history by studying horn fly development.

Improved Sentinel Method for Surveillance and Collection of Filth Fly Parasitoids.

Parasitoids are important natural enemies of house flies and other muscoid flies. The two most commonly used methods for collecting fly parasitoids from the field have distinct advantages and disadvantages. Collections of wild puparia depend on the ability to find puparia in sufficient numbers and are prone to localized distortions in relative species abundance because of the overrepresentation of samples from hot spots of fly larval activity. Placement and retrieval of sentinel puparia is convenient and allows consistent sampling over time but is strongly biased in favor of Muscidifurax spp. over Spalangia spp. An improved sentinel method is described that combines some of the advantages of these two methods. Fly medium containing larvae is placed in containers, topped with a screen mesh bag of puparia, and placed in vertebrate-proof wire cages. Cages are placed at sites of actual or potential fly breeding and retrieved 3-7 d later. The modified method collected species profiles that more closely resembled those of collections of wild puparia than those from sentinel pupal bags. A method is also described for isolating puparia individually in 96-well tissue culture plates for parasitoid emergence. Use of the plate method provided a substantial saving of time and labor over the use of individual gelatin capsules for pupal isolation. Puparia from the collections that were housed individually in the wells of tissue culture plates had a higher proportion of emerged Spalangia species than puparia that were held in groups.

Haematobium irritans and Haematobium titillans as potential vectors of Parabronema skrjabini in camels (Camelus bactrianus) in Inner Mongolia, China.

Parabronema skrjabini is one of the most harmful nematodes to camels and is responsible for economic losses in animal husbandry industry. There is an urgent need for in-depth studies of potential vectors of the nematode due to its scant regarding information. As previous studies indicated that flies may be the vectors of P. skrjabini, we captured flies in the main camel-producing areas of Inner Mongolia. After autopsy of the specimens of two species of horn flies, we observed the morphology of the suspected nematode larvae found in them. Internal transcribed spacer ribosomal-DNA gene sequences were considered the best candidate to confirm the species of the larvae found. Our results showed that the homology compared with P. skrjabini was 99.5% in GenBank. Subsequently, we preliminarily identified two species of horn flies through morphological observation and then sequenced the mitochondrial-DNA-gene cytochrome oxidase subunit I obtained from two species of horn flies, with 100 and 99.2% similarity to sequences deposited in GenBank, respectively. Thus, we identified Haematobia titillans and Haematobia irritans and provided evidence for their potential role as vectors of parabronemosis. Our study provides reference for future research on the life history of the nematode and the vectors of parabronemosis.

DNA of Theileria orientalis, T. equi and T. capreoli in stable flies (Stomoxys calcitrans).

BACKGROUND: From a veterinary-medical point of view, the stable fly, Stomoxys calcitrans, is perhaps the economically most important blood-sucking muscoid fly species (Diptera: Muscidae), owing to its worldwide occurrence, frequently high local abundance, direct harm caused to livestock, pet animals and humans, as well as its vector role. Considering the latter in the context of protozoan parasites, the stable fly is a mechanical vector of trypanosomes and Besnoitia besnoiti. However, its role as a vector of piroplasms appears to be seldom studied, despite old data suggesting mechanical transmission of babesiae by dipteran flies. METHODS: In this study 395 stable flies (and one Haematobia stimulans) were collected at a cattle farm with known history of bovine theileriosis, and at further nine, randomly chosen locations in Hungary. These flies were separated according to sex (30 of them also cut into two parts: the head with mouthparts and the thorax-abdomen), followed by individual DNA extraction, then screening for piroplasms by PCR and sequencing. RESULTS: In stable flies, Theileria orientalis and T. capreoli were identified at the cattle farm and T. equi was identified in three other locations. At the cattle farm, significantly more male stable flies carried piroplasm DNA than females. There was no significant difference between the ratio of PCR-positive flies between the stable (void of cattle for at least two hours) and the pen on the pasture with cattle at the time of sampling. Among dissected flies (29 S. calcitrans and 1 H. stimulans), exclusively the thoracic-abdominal parts were PCR-positive, whereas the head and mouthparts remained negative. CONCLUSIONS: Theileria DNA is detectable in stable flies, in the case of T. orientalis at least for two hours after blood-feeding, and in the case of T. capreoli also in the absence of infected hosts (i.e. roe deer). Male flies rather than females, and thoracic-abdominal (most likely crop) contents rather than mouthparts may pose a risk of mechanical transmission. These data suggest that it is worth to study further the vector role of stable flies in the epidemiology of theilerioses, in which not the immediate, but rather the delayed type transmission seems possible.

Effect of Fluctuating High Temperatures on House Flies (Diptera: Muscidae) and Their Principal Parasitoids (Muscidifurax spp. and Spalangia spp. [Hymenoptera: Pteromalidae]) From the United States.

Colonies of house flies (Musca domestica L. [Diptera: Muscidae]) and four species of parasitoids (Muscidifurax raptor Girault and Sanders, Muscidifurax zaraptor Kogan and Legner, Spalangia cameroni Perkins and Spalangia endius Walker) were established by making collections from dairy farms near Bell, FL, Beatrice, NE, Minneapolis, MN, and San Jacinto, CA. Colonies were assessed for heat tolerance by comparing life history parameters at 25-27°C and fluctuating hot (26.7-41.7°C) temperatures. Muscidifurax raptor, S. cameroni, and S. endius produced 24-28% as many progeny under hot conditions as at 25°C. Colonies of M. zaraptor were more heat-tolerant and produced an average 46.9% as many progeny under the hot regime compared with moderate conditions. There was little evidence for higher heat tolerance in parasitoid populations from historically hot locations (CA desert and FL). Colonies of M. raptor and S. endius that had been in culture for 24 yr were the least heat-tolerant with regard to progeny production. House flies collected from the same locations varied little in longevity, fecundity, or egg-to-adult survival under either hot or moderate regimes. Flies reared under hot conditions laid about half as many eggs (89/female) and had about half the egg-adult survival rate (47.3%) under hot compared with moderate conditions, indicating that heat stress had less effect on flies than on all of the parasitoids except M. zaraptor. An attempt to select for heat tolerance in flies by subjecting them to incremental increases in rearing temperatures for 20 generations resulted in little change in tolerance among the selected flies.

Phenotypic plasticity more essential to maintaining variation in host-attachment behaviour than evolutionary trade-offs in a facultatively parasitic mite.

A prevailing hypothesis for the evolution of parasitism posits that the fitness benefits gained from parasitic activity results in selection for and fixation of parasitic strategies. Despite the potential fitness advantage of parasitism, facultative parasites continue to exhibit genetic variation in parasitic behaviour in nature. We hypothesized that evolutionary trade-offs associated with parasitic host-attachment behaviour maintain natural variation observed in attachment behaviour. In this study, we used replicate lines of a facultatively parasitic mite, previously selected for increased host-attachment behaviour to test whether increased attachment trades off with mite fecundity and longevity, as well as the phenotypic plasticity of attachment. We also tested for potential correlated changes in mite morphology. To test for context-dependent trade-offs, mite fecundity and longevity were assayed in the presence or absence of a host. Our results show that selected and control mites exhibited similar fecundities, longevities, attachment plasticities and morphologies, which did not provide evidence for life history trade-offs associated with increased attachment. Surprisingly, phenotypic plasticity in attachment was maintained despite directional selection on the trait, which suggests that phenotypic plasticity likely plays an important role in maintaining attachment variation in natural populations of this facultative parasite.

Molecular identification of bloodmeal sources and trypanosomes in Glossina spp., Tabanus spp. and Stomoxys spp. trapped on cattle farm settlements in southwest Nigeria.

The interactions of host, vector and parasite in bovine trypanosomiasis transmission cycles in southwest Nigeria are not yet well understood. Trypanosoma (Trypanosomatida: Trypanosomatidae) species infection prevalences and bloodmeal sources were determined in transmitting vectors of the genera Glossina (Diptera: Glossinidae), Tabanus (Diptera: Tabanidae) and Stomoxys (Diptera: Muscidae) collected using Nzi traps in cattle settlements in southwest Nigeria. Sequenced cytochrome B mitochondrial DNA segments obtained from vector digestive tracts identified bloodmeal sources from eight host species, namely human, cattle, hippopotamus, giraffe, gazelle, spotted hyena, long-tailed rat and one unidentified species. Overall, 71.1% [95% confidence interval (CI) 63.0-78.1], 33.3% (95% CI 21.9-47.0) and 22.2% (95% CI 16.2-29.9), respectively, of Glossina, Tabanus and Stomoxys flies were positive for trypanosomes. The observed trypanosome species were Trypanosoma vivax, Trypanosoma congolense, Trypanosoma brucei, Trypanosoma evansi, Trypanosoma simiae and Trypanosoma godfreyi. Trypanosome DNA was more prevalent in tsetse (34.8% Tr. vivax, 51.1% Tr. b. brucei, 5.2% Tr. congolense, 4.4% Tr. simiae and 24.4% mixed infections) than in other flies and the main determinants in all flies were seasonal factors and host availability. To the best of the present group's knowledge, this is the first report of Trypanosoma species in Tabanus and Stomoxys flies in Nigeria. It indicates that vector control programmes should always consider biting flies along with tsetse flies in the control of human and animal trypanosomiasis.

Potential of entomopathogenic nematodes of the genus Heterorhabditis for the control of Stomoxys calcitrans (Diptera: Muscidae) / Potencial de nematoides entomopatogênicos do gênero Heterorhabditis para o controle de Stomoxys calcitrans (Diptera: Muscidae)

Abstract The present study verified the pathogenic potential of entomopathogenic nematodes (EPNs) of the genus Heterorhabditis (Heterorhabditis bacteriophora, isolate HP88 and Heterorhabditis baujardi isolate LPP7) to immature stages of Stomoxys calcitrans in the laboratory. All EPN concentrations of the H. bacteriophora HP88 strain caused mean larval mortality greater than 90% after four days. Higher concentrations of the H. baujardi LPP7 isolate (≥50 EPNs/larva) eliminated more than 70% of larvae after six days with the concentration 200 EPNs/larva reaching mortality levels of 93.3%. The larval mortality at all concentrations of EPNs (25, 50, 100, 150, and 200 EPNs/larva) for both strains was significant (p<0.05) when compared to the respective control groups. Concentrations of H. bacteriophora HP88 yielded an LC50 of 0.36 EPN/larva and LC90 of 29.1; while H. baujardi LPP7 yielded an LC50 of 39.85 and LC90 of 239.18. H. bacteriophora HP88 provided greater inhibition of the emergence of adults when compared to the response obtained with H. baujardi LPP7. EPNs did not cause considerable mortality when applied directly to pupae. The set of observed results suggests that the EPNs of the genus Heterorhabditis, isolates HP88 and LPP7, are a promising alternative in the control of the stable fly.

Resumo O presente estudo verificou o potencial patogênico de nematoides (NEP) do gênero Heterorhabditis (H. bacteriophora - isolado HP88 e H. baujardi - isolado LPP7) para estágios imaturos de Stomoxys calcitrans em laboratório. Todas as concentrações de NEPs da cepa HP88 causaram mortalidade larval média maior que 90% após quatro dias. Concentrações mais elevadas da cepa LPP7 (≥50 NEPs/larva) eliminaram mais de 70% após seis dias, com a concentração 200 NEPs/larva atingindo níveis de 93,3%. A mortalidade larval em todas as concentrações de NEPs (25, 50, 100, 150, and 200 NEPs/larva) para ambas as cepas foi significativa (p<0,05), quando comparadas aos respectivos grupos controle. Concentrações de H. bacteriophora HP88 obtiveram LC50 de 0,36 NEP/larva e LC90 de 29,1; enquanto H. baujardi LPP7 obteve LC50 de 39,85 e LC90 de 239,18. O isolado H. bacteriophora HP88 propiciou maior inibição da emergência de adultos, quando comparado à resposta obtida com H. baujardi LPP7. NEPs não provocaram mortalidade considerável quando aplicados diretamente sobre pupas. O conjunto de resultados observados indica os nematoides entomopatogênicos do gênero Heterorhabditis (estirpes HP88 e LPP7) como uma alternativa promissora no controle da mosca dos estábulos.

Potential of entomopathogenic nematodes of the genus Heterorhabditis for the control of Stomoxys calcitrans (Diptera: Muscidae).

The present study verified the pathogenic potential of entomopathogenic nematodes (EPNs) of the genus Heterorhabditis (Heterorhabditis bacteriophora, isolate HP88 and Heterorhabditis baujardi isolate LPP7) to immature stages of Stomoxys calcitrans in the laboratory. All EPN concentrations of the H. bacteriophora HP88 strain caused mean larval mortality greater than 90% after four days. Higher concentrations of the H. baujardi LPP7 isolate (≥50 EPNs/larva) eliminated more than 70% of larvae after six days with the concentration 200 EPNs/larva reaching mortality levels of 93.3%. The larval mortality at all concentrations of EPNs (25, 50, 100, 150, and 200 EPNs/larva) for both strains was significant (p<0.05) when compared to the respective control groups. Concentrations of H. bacteriophora HP88 yielded an LC50 of 0.36 EPN/larva and LC90 of 29.1; while H. baujardi LPP7 yielded an LC50 of 39.85 and LC90 of 239.18. H. bacteriophora HP88 provided greater inhibition of the emergence of adults when compared to the response obtained with H. baujardi LPP7. EPNs did not cause considerable mortality when applied directly to pupae. The set of observed results suggests that the EPNs of the genus Heterorhabditis, isolates HP88 and LPP7, are a promising alternative in the control of the stable fly.

Differential Haematobia irritans infestation levels in beef cattle raised in silvopastoral and conventional pasture systems.

The use of silvopastoral systems (SPS) can be a good alternative to reduce the environmental impacts of livestock breeding in Brazil. One of the reasons for its scarce adoption is the lack of information on health and productivity of cattle raised under these conditions. The experiment reported here was designed to compare the infestation by external parasites - the cattle tick (Rhipicephalus microplus), horn fly (Haematobia irritans), and larvae of the botfly (Dermatobia hominis) - in beef cattle raised in a SPS and a conventional pasture system (CPS), evaluated for 24 months. Data on air and soil temperature, solar radiation, wind incidence and water balance were used to characterize the SPS and CPS. R. microplus adult females and D. hominis larvae were counted on the body of each animal to determine the parasites burdens, but we did not find significant differences between the two systems. Horn flies counts on animals' body, and analysis of the horn fly and its pupal parasitoids associated with the dung pats were obtained in the two systems. Horn fly infestation was significantly lower (p=0.01) in the SPS (13.17±3.46) in comparison with the CPS (24.02±4.43). In SPS and CPS, respectively, the mean densities of pupae of H. irritansin dung pats were 9.8 and 10.7; the mean density of adults of H. irritans, 3.7 and 3.5; and the density of its pupal parasitoids were 20.5 and 5.4. The effect of production system was significant (p<0.05) only for the occurrence of pupal parasitoids of the horn fly, where the greatest occurrences of these natural enemies were in the SPS. These data indicate that natural enemies were able to control, at least partially, the horn fly populations in the cattle.

The larval development of Habronema muscae (Nematoda: Habronematidae) affects its intermediate host, Musca domestica (Diptera: Muscidae).

Although the life cycle of the equid stomach parasite Habronema muscae was disclosed more than 100 years ago, little is known about the effect of the developing nematode larvae in its intermediate host, Musca domestica. In a series of experiments, freshly hatched M. domestica larvae were exposed to H. muscae eggs contained in a faecal sample of a naturally infected horse. In daily intervals, 50 fly larvae were removed and transferred on a parasite-free larval rearing medium where they completed their development. Hatched flies were examined for the presence of Habronema third-stage larvae. In two subsequent control groups, flies spend their entire larval life in contaminated horse faeces and in a parasite-free larval rearing medium, respectively. Out of the 700 fly larvae used in the infection experiments, 304 developed into adult flies of which 281 were infected. The average nematode larval burden rose from 3.6 in the group with the shortest exposure to more than 25 in the groups with the longest exposure. The proportion of larvae that developed into the adult insect fell from 82 % in the uninfected control group to 27 % in the positive control group. The pupae of the positive control group were smaller and lighter than those of the uninfected control group. Lower pupal size and weight in the positive control group as well as a lower insect developing rate might be attributed to the destruction of adipose cells in the maggots by Habronema larvae.

First Record of the Larval Endoparasitoid Tachinaephagus zealandicus (Hymenoptera: Encyrtidae) From Fly Hosts Developing on Burnt Remains and in Pennsylvania.

Adults of the gregarious larval endoparasitoid Tachinaephagus zealandicus Ashmead (Hymenoptera: Encyrtidae) were collected from two species of carrion-inhabiting flies, Phormia regina Meigen (Diptera: Calliphoridae) and Hydrotaea aenescens Wiedemann (Muscidae), associated with the burnt remains of a domesticated pig, Sus scrofa L. (Artiodactyla: Suidae), during late summer in south central Pennsylvania. This represents the first reported occurrence of the wasp in the state and only the second in the mid-Atlantic region. Collection from P. regina is a new host record for T. zealandicus and also the first for this parasitoid being attracted to flies on burnt remains.

Association of Myianoetus muscarum (Acari: Histiostomatidae) With Synthesiomyia nudiseta (Wulp) (Diptera: Muscidae) on Human Remains.

Synthesiomyia nudiseta (Wulp) (Diptera: Muscidae) was identified during the course of three indoor medicolegal forensic entomology investigations in the state of Texas, one in 2011 from Hayes County, TX, and two in 2015 from Harris County, TX. In all cases, mites were found in association with the sample and subsequently identified as Myianoetus muscarum (L., 1758) (Acariformes: Histiostomatidae). This report represents the first records of a mite associated with S. nudiseta in the continental United States. In particular, this association is believed to be of potential future value in forensic investigations, as it lends new insight into the community structure of colonizers on human remains in indoor environments.

Diversity and Abundance of House Fly Pupal Parasitoids in Israel, with First Records of Two Spalangia Species.

Filth flies (Diptera: Muscidae), particularly the house fly, Musca domestica L., are global pests of livestock production. In this study, we characterized the fauna of house fly pupal parasitoids in Israel and identified factors affecting their diversity and abundance. The study, which included one round of sampling during the fall of 2013 and another round of sampling in the spring of 2014, encompassed 26 locations of common fly-breeding habitats: dairy, egg-laying, and goat farms throughout Israel. Nine parasitoid species were found: Spalangia cameroni Perkins, Spalangia endius Walker, Spalangia drosophilae Ashmead, Spalangia gemina Boucek, Spalangia nigroaenea Curtis, Pachycrepoideus vindemmiae Rondani, Muscidifurax raptor Girault and Sanders, Muscidifurax zaraptor Kogan & Legner (all Hymenoptera: Pteromalidae), and Dirhinus giffardii Silvestri (Hymenoptera: Chalcididae). This is the first record of S. gemina from the Palearctic, as well as the first record of S. drosophilae from the Middle East. The composition and relative abundance of parasitoid species varied markedly among localities, climatic regions (Mediterranean vs. desert), habitat types (dairy vs. egg-laying vs. goat farm), and seasons. Overall, parasitoid richness in egg-laying farms was two- and sevenfold higher than in dairy and goat farms, respectively, and three times higher in Mediterranean than desert climate. The significance and implications of our results for inundative biological control programs of filth flies are discussed.

Temperature-Dependent Functional Response of Spalangia cameroni (Hymenoptera: Pteromalidae), a Parasitoid of Stomoxys calcitrans (Diptera: Muscidae).

The effects of host density, temperature, and burial depths on the functional response of the synovigenic parasitoid Spalangia cameroni (Perkins) attacking pupae of the stable fly Stomoxys calcitrans (L.) were examined. Five temperatures (15, 20, 25, 30, and 35°C), six host densities (2, 4, 8, 16, 32, and 64 pupae per 19.64 cm(2)), and three burial depths in whole wheat grains (0, 8, and 16 cm) were used. It was demonstrated that temperature had a profound effect on the functional response, whereas burial depths of the pupae played a minor role. For all temperatures, the functional response was of type 2 with an upper asymptote that depends on the temperature. Data were fitted by a functional response model where only the maximum attack rate is temperature dependent. The model shows that the optimal temperature for S. cameroni for attacking S. calcitrans pupae is 28.6°C, where the maximum attack rate is 20.2 pupae day(-1). The lower and upper temperature thresholds for attacks were found as 8.1 and 36.6°C, respectively. The temperature range for attacks that resulted in successful parasitism was narrower, namely, 15.0 and 35.5°C. Maximum rate of successful parasitism was estimated to be 18.2 progeny day(-1) occurring at 27.9°C. The proportion of attacks resulting in successful parasitism increased steeply with temperatures >15°C and declined steeply at temperatures >30°C.

Comparison of the Olfactory Preferences of Four of Filth Fly Pupal Parasitoid Species (Hymenoptera: Pteromalidae) for Hosts in Equine and Bovine Manure.

House flies (Musca domestica L.) and stable flies (Stomoxys calcitrans (L.)) (Diptera: Muscidae) are common pests in equine and cattle facilities. Pupal parasitoids, primarily in the genera Spalangia and Muscidifurax (Hymenoptera: Pteromalidae), can be purchased for biological control of these flies. However, little is known about the host-habitat preferences associated with host-seeking by these parasitoids. The preferences of two Spalangia and two Muscidifurax species to odors associated with house fly hosts in equine and bovine manure were investigated in the laboratory using a Y-tube olfactometer. Odor stimuli from manure without developing flies, third-instar house flies in manure, and fly host puparia in manure were evaluated. In choice tests, S. cameroni and S. endius were strongly attracted to odor associated with equine manure against clean air. Although S. cameroni was attracted to all bovine manure-containing treatments against clean air, S. endius was only attracted to the bovine manure with third-instar flies. There were no significant differences between the Spalangia species in odor responses. Neither Muscidifurax species were attracted to equine manure treatments and were only attracted to the bovine manure with puparia over clean air. In manure comparison studies, bovine treatments with developing flies were more attractive than the equivalent equine treatments to both Muscidifurax species The data suggest that coexistence between the competing pteromalid parasitoids might be promoted by different host-seeking behaviors. Additionally, manure preferences may indicate parasitoid suitability for releases on different livestock and equine facilities.

Effect of Mutual Interference on the Ability of Spalangia cameroni (Hymenoptera: Pteromalidae) to Attack and Parasitize Pupae of Stomoxys calcitrans (Diptera: Muscidae).

We investigated the effect of mutual interference on the attack efficiency and the rate of successful parasitism on the parasitoid Spalangia cameroni (Perkins) attacking pupae of the stable fly Stomoxys calcitrans (L.). Female parasitoids (2, 4, 8, 16, or 32) were exposed to 100 fly pupae during 24 h. The number of pupae that were attacked and the number successfully parasitized increased with the parasitoid density and reached a maximum of ∼70 and 50, respectively. Parasitoid-induced mortality (PIM) was about 20 pupae, irrespective of parasitoid density. The per capita rates of attack, successful parasitism and parasitoid-induced mortality declined monotonously with parasitoid density. Progeny sex ratio was female biased for all parasitoid densities, but declined significantly with increasing parasitoid density from ∼70% females at the lowest density to ∼60% at the highest. Mutual interference was incorporated into a functional response model to predict the attack rate and the rate of successful parasitism at different temperatures, host densities and parasitoid densities. The model explained 93.5% of the variation in the observed number of attacked pupae and 91.5% of the variation in the number of successfully parasitized pupae. The model predicts that increasing parasitoid densities will increase the percentage of killed hosts, but only up to a certain density. Above this density, a further increase in parasitoid abundance will actually lead to a decline in the percentage parasitism. These findings may have some implications for using S. cameroni in biological control against flies using inundative releases.

Linear Dispersal of the Filth Fly Parasitoid Spalangia cameroni (Hymenoptera: Pteromalidae) and Parasitism of Hosts at Increasing Distances.

Release of parasitic wasps (Hymenoptera: Pteromalidae) as biological control agents for house flies and stable flies in livestock confinements has had variable success. In part, this may reflect a lack of knowledge regarding the optimal distance to be used between parasitoid release stations. In the current study, we assessed the effect of linear distance on host parasitism by the wasp Spalangia cameroni Perkins. In open fields at distances ranging from 1 m to 60 m from a central point, house fly puparia were placed in a mixture of pine shavings soiled with equine manure, urine, and alfalfa hay. Releases of S. cameroni then were made using a 5:1 host: parasitoid ratio. Host pupae were parasitized at all distances, with the highest rate of total parasitism (68.9%) recorded ≤ 5 m from the release site. Analyses of results using non-linear and linear models suggest that S. cameroni should be released in close proximity to host development areas. Additionally, releases may not be suitable in pasture situations where long-distance flight is required for control. However, further testing is needed to examine the effect of density-dependent dispersal and diffusion of S. cameroni.