The prevalence of Stephanofilaria sp. in buffalo fly, Haematobia irritans exigua, in Central Queensland

Abstract  Buffalo fly ( Haematobia irritans exigua ) infestations of cattle are associated with characteristic lesions, the initial cause of which has been attributed to a filarial nematode of the genus Stephanofilaria , for which the fly acts as a vector. Survey work in the 1980s estimated the prevalence of microfilaria in female buffalo fly in Queensland at 2.91%. Since then no information has been published and the current prevalence of microfilarial infection in buffalo fly is not known. Buffalo fly were collected from four geographically distinct sites in Central Queensland in mid-summer 2004 and were dissected to estimate Stephanofilaria sp. infection rates . Larval stages of the nematodes were recovered from female flies from all four sites and the percentage of female flies from which nematodes were recovered ranged from 29% to 57%. The average number of larvae recovered from infected female flies ranged from 1.25 to 1.75. Whereas no infected male flies were found from Sites 2–4, larvae were recovered from 43% of male flies collected at Site 1. This high prevalence of filarial infection in buffalo flies implies a correspondingly high level of transmission to cattle in Central Queensland.     

Competitive Superiority of Early Acting Species: A Case Study of Opiine Fruit Fly Parasitoids

Koinobiont parasitoids that attack an early host stage may have an advantage in suppressing competing parasitoids that attack later stages of the same host. We examined the competitive interaction between the two most important parasitoids of tephritid fruit flies in Hawaii, Fopius arisanus (Sonan), and Diachasmimorpha longicaudata (Ashmead) (Hymenoptera: Braconidae). The former species attacks host eggs while the latter attacks host larvae, and both species emerge as adults from the host puparia. F. arisanus physiologically suppressed egg development of D. longicaudata. Over 90% of D. longicaudata eggs died in the presence of F. arisanus larvae within host larvae of either the Mediterranean fruit fly, Ceratitis capitata (Wiedemann) or the oriental fruit fly, Bactrocera dorsalis (Hendel). D. longicaudata appeared not to discriminate against hosts previously parasitized by F. arisanus. The mechanism that F. arisanus uses to eliminate D. longicaudata is similar to that which it employs to eliminate five other larval fruit fly parasitoids so far reported in Hawaii. This suggests that there is a broad competitive superiority of the early acting species in fruit fly parasitoids. We discuss the implication of this in relation to future biological control introductions against tephritid fruit flies.     

Leishmania infantum xenodiagnosis from vertically infected dogs reveals significant skin tropism

BackgroundDogs are the primary reservoir for human visceral leishmaniasis due to Leishmania infantum. Phlebotomine sand flies maintain zoonotic transmission of parasites between dogs and humans. A subset of dogs is infected transplacentally during gestation, but at what stage of the clinical spectrum vertically infected dogs contribute to the infected sand fly pool is unknown.Methodology/Principal findingsWe examined infectiousness of dogs vertically infected with L. infantum from multiple clinical states to the vector Lutzomyia longipalpis using xenodiagnosis and found that vertically infected dogs were infectious to sand flies at differing rates. Dogs with mild to moderate disease showed significantly higher transmission to the vector than dogs with subclinical or severe disease. We documented a substantial parasite burden in the skin of vertically infected dogs by RT-qPCR, despite these dogs not having received intradermal parasites via sand flies. There was a highly significant correlation between skin parasite burden at the feeding site and sand fly parasite uptake. This suggests dogs with high skin parasite burden contribute the most to the infected sand fly pool. Although skin parasite load and parasitemia correlated with one another, the average parasite number detected in skin was significantly higher compared to blood in matched subjects. Thus, dermal resident parasites were infectious to sand flies from dogs without detectable parasitemia.Conclusions/SignificanceTogether, our data implicate skin parasite burden and earlier clinical status as stronger indicators of outward transmission potential than blood parasite burden. Our studies of a population of dogs without vector transmission highlights the need to consider canine vertical transmission in surveillance and prevention strategies.     

Field cage assessment of interference among insects attacking seed heads of spotted and diffuse knapweed

Field studies were conducted to determine the competitive interactions between introduced biological control agents that attack the seed heads of spotted knapweed (Centaurea stoebe ssp. micranthos) and diffuse knapweed (Centaurea diffusa). Two weevils, Bangasternus fausti and Larinus minutus (Coleoptera: Curculionidae), were each paired with the previously established fly, Urophora affinis (Diptera: Tephritidae). Each species was released either alone or in pair-wise combinations inside screen cages placed over existing knapweed plants at six field sites in Montana and one in Oregon. Larinus minutus produced almost three times as many progeny on diffuse knapweed as on spotted knapweed. Larinus minutus reproduction was not affected by competition with U. affinis, but U. affinis reproduction was reduced by the presence of L. minutus (by 71% on spotted and 77% on diffuse knapweed). Bangasternus fausti reproduction generally was not affected by competition with U. affinis, nor was U. affinis affected by B. fausti on either host plant. There were extremely few cases of successful production of both weevil and fly in the same capitulum, which was probably because weevil larvae consume the developing flies. Both weevils increased the total proportion of seed heads infested on diffuse knapweed, and B. fausti increased it on spotted knapweed. However, the release of either weevil did not significantly further reduce seed production on either plant. The results and experimental design are discussed in light of the subsequent establishment and impact of these agents.     

Dynamics of Haematobia irritans irritans (Diptera: Muscidae) infestation on Nelore cattle in the Pantanal, Brazil

From June 1993 to May 1995, horn fly counts were conducted twice a month on untreated Nelore cattle raised extensively in the Pantanal. Horn fly population showed a bimodal fluctuation and peaks were observed every year after the beginning (November/December) and at the end (May/June) of the rainy season, which coincided with mid-late spring and mid-late fall, respectively. Horn flies were present on cattle throughout the year in at least 64% of the animals. Mean horn fly numbers on animals did not exceed 85 flies/cow during peaks and were under 35 flies/cow in most of the remaining periods. The highest infestations (population peaks) were short and dropped suddenly within two weeks. Less than 15% of the animals in both herds could be considered as "fly-susceptible" - showing consistently higher infestations, or "fly-resistant" - showing consistently lower infestations.     

Thelazia lacrymalis in horses in Kentucky and observations on the face fly (Musca autumnalis) as a probable intermediate host.

Eyes from 114 (30.3%) of 376 dead horses, examined from 3 April 1975 to 3 April 1976, were naturally infected with adult Thelazia lacrymalis; 1 horse was also infected with 1 male Thelazia skrijabini. Adult T. lacrymalis from dead horses were successfully transferred mechanically to the eyes of 3 of 4 Shetland ponies raised helminth-free. Larvae from gravid female T. lacrymalis underwent development in experimentally infected, laboratory-raised face flies (Musca autumnalis) and third-stage larvae ranging from 1.82 to 2.94 mm in total length were recovered at 12 to 15 days postexposure. A total of 866 naturally occurring face flies were collected from the head region of horses. Twelve of the face flies harbored larval stages of Thelazia spp. One of the larvae resembled third-stage T. lacrymalis that were recovered from the experimentally infected, laboratory-raised face flies. Introduction of 3 third-stage larvae from 1 face fly onto the cornea of a pony raised helminth-free resulted in the recovery of 1 male T. skrjabini 242 days later. In addition to the eyeworm larvae, other parasites recovered from the face flies included Heterotylenchus autumnalis, hypopi of astigmatid mites and a first instar beetle (Coleoptera: Rhipiphoridae). Data from these investigations indicate the likelihood that face flies are an intermediate host for T. lacrymalis and probably other species of Thelazia in this part of the country.     

Dermacentor variabilis: acquired resistance to ticks in BALB/c mice

To assess the ability of Dermacentor variabilis larvae to feed on mice during repeated infestations, known numbers of larvae were confined to the skin of BALB/c mice within plastic capsules, and different skin sites were used in four successive infestations. In secondary infestations larval feeding appeared to be enhanced, but in subsequent infestations the mice expressed acquired tick resistance, manifested by reduction in numbers of engorged larvae and in weights of fed larvae. Resistance, once established, was still demonstrable four weeks after the previous infestation. Histological examination of skin biopsies from BALB/c mice undergoing repeated infestations revealed slight reactions to primary infestations and increasingly severe responses in subsequent infestations. The latter reactions included prominent participation of mast cells and eosinophils, many of which were degranulated close to tick bite sites. It is suggested that these cells might play a role in the expression of murine tick resistance.     

Field application of an insect virus in the Mekong Delta: Effects of a Vietnamese nucleopolyhedrovirus on Spodoptera litura (Lepidoptera: Noctuidae) and its parasitic natural enemies

A new Vietnamese isolate of Spodoptera litura nucleopolyhedrovirus (NPV) was applied in local soybean fields, and the effect of its application on S. litura larvae was examined. The virus was propagated in vivo and a crude extract was prepared for spraying at high and low doses (1.7×10⁸ and 3.3×10⁷ occlusion bodies/m², respectively). The percentage of larvae infected with NPV increased from 22.2% on the day before NPV application (Day 0) to 50.8% (Day 6) in the high dose treatment plot, and from 7.9% (Day 0) to 35.7% (Day 6) in the low-dose plot. Microsporidium sp. was observed as another major pathogen of S. litura larvae. Three dominant parasitic natural enemies were found in S. litura larvae: Microplitis manilae (Braconidae), Chelonus sp. (Braconidae), and Peribaea orbata (Tachinidae). The fate of parasitoids developing within virus- and Microsporidium- infected hosts differed between these three parasitoids; more Chelonus sp. emerging from infected hosts died during their larval stage before spinning cocoons, or failed to reach adult eclosion, than did P. orbata. This suggests that the impact of virus application on the survival of parasitoids varies from species to species.     

Cold treatment for guava fruits infested with oriental fruit fly,Bactrocera dorsalis (Diptera: Tephritidae)

Guava (Psidium guajava Linnaeus) is an important horticulture crop in tropical and subtropical regions. However, guava transportation to the fruit market is regulated due to the risk of fruit fly infestations. The oriental fruit fly [Bactrocera dorsalis (Hendel), Tephritidae, Diptera], which damages numerous horticulture fruits, is the primary pest in guava. For fruit export, phytosanitary treatment is necessary to prevent the potential oriental fruit fly infestations through pest-infested fruits. In this study, cold treatment was administered to eliminate oriental fruit fly infestations in guava fruits. The optimal egg inoculation density was 200 eggs per fruit, and third instar larvae displayed the highest cold treatment tolerance. On assessing the efficacy of cold treatment, none of the third instar larvae in the fruits survived at a constant fruit core temperature of 0.5–1 °C for 11 days. A confirmatory test revealed that on maintaining a fruit core temperature of 0.5–1 °C for 12 days, 100% mortality was achieved among the fruit flies. In fruit quality assessment, guava fruits exposed to cold treatment still maintained their market value. Thus, cold treatment effectively disinfested guava fruits with possible fruit fly infestations, making it a viable quarantine treatment for guava fruits prior to their export.     

Acetaldehyde detoxification mechanisms in Drosophila melanogaster adults involving aldehyde dehydrogenase (ALDH) and alcohol dehydrogenase (ADH) enzymes

The mechanism of acetaldehyde detoxification in Drosophila melanogaster adults has been studied by comparing physiological in vitro and in vivo data. ADH⁺ and ADH⁻ flies, both lacking aldehyde dehydrogenase activity from ADH (ALDH^ADH, ALDH (ALDH) or both enzymes were exposed to acetaldehyde or ethanol, and the toxicity and internal accumulation of both compounds were determined. Acetaldehyde was extremely lethal for flies whose ALDH activity had been inhibited by cyanamide, though acetaldehyde was effectively detoxified by flies whose ALDH^ADH activity had been inhibited by acetone. After exposure to acetaldehyde, both acetaldehyde and ethanol rapidly accumulated in flies lacking ALDH activity, but not in flies lacking ALDH^ADH activity. However, ethanol but not acetaldehyde quickly accumulated in flies lacking ALDH activity after exposure to ethanol. Our results provide in vivo evidence that, as opposed to larvae, in D. melanogaster adults acetaldehyde is mainly oxidized into acetate by means of ALDH enzymes. However, the reducing activity of the ADH enzyme, which transforms acetaldehyde into ethanol, also plays an essential role in the detoxification of acetaldehyde. Differences in ALDH activity might be important to explain the differences in ethanol tolerance found in natural populations.     

Parasitism of Heterotylenchus autumnalis Nickle (Nematoda: Sphaerulariidae) to the Face Fly,Musca autumnalis De Geer (Diptera: Muscidae)

New information is reported on the parasitism of Heterotylenchus autumnalis upon its principal known host, Musca autumnalis. Black to brown spots are produced on the cuticle of all infected host larvae where the nematode penetrated. The principal damage to the host is castration of the female. In laboratory tests nematode larvae were not infective and did not leave the hosts before the female fly was 1 1 days old. Nematode larvae removed from infected male flies infected other hosts, but it is believed that in nature these larvae are unable to leave the host.     

Susceptibility of black soldier fly (Diptera: Stratiomyidae) larvae and adults to four insecticides

Dosage-mortality regressions were determined for black soldier fly, Hermetia illucens (L.), larvae fed cyromazine or pyriproxifen treated media. Cyromazine LC50 for larvae dying before becoming prepupae ranged from 0.25 to 0.28 ppm with dosage-mortality regression slopes between 5.79 and 12.04. Cyromazine LC50s for larvae dying before emergence ranged from 0.13 to 0.19 ppm with dosage-mortality regression slopes between 3.94 and 7.69. Pyriproxifen dosage-mortality regressions were not generated for larvae failing to become prepupae since <32% mortality was recorded at the highest concentration of 1,857 ppm. LC50s for larvae failing to become adults ranged from 0.10 to 0.12 ppm with dosage mortality-regression slopes between 1.67 and 2.32. Lambda-cyhalothrin and permethrin dosage-mortality regressions were determined for wild adult black soldier flies and house flies, Musca domestica L., and for susceptible house flies. Our results indicate that the wild house fly, unlike the black soldier fly, population was highly resistant to each of these pyrethroids. Regression slopes for black soldier flies exposed to lambda-cyhalothrin were twice as steep as those determined for the wild house fly strain. Accordingly, LC50s for the black soldier fly and susceptible house fly were 10- to 30-fold lower than those determined for wild house flies. The differential sensitivity between wild black soldier flies and house flies might be due to behavioral differences. Adult house flies usually remain in animal facilities with the possibility of every adult receiving pesticide exposure, while black soldier fly adults are typically present only during emergence and oviposition thereby limiting their exposure.     

Use of a monoclonal antibody to ovine IgE for fly strike studies in sheep

A new monoclonal raised against sheep IgE was used to examine sera and wound exudates from sheep which had been struck by Lucilia cuprina in the field. The antibody was also used to detect the presence of IgE in sera and skin sections from sheep which had been artificially infected with fly larvae 3 times. Neither total, nor L. cuprina specific circulating IgE could be detected in serum or wound exudates from struck sheep. Cell bound IgE was, however, identified by the monoclonal in skin sections from struck sheep and from a control sheep which had not been struck. No difference in the number of IgE positive cells was observed between the control and 2 of the 3 artificially infected sheep, and none of the latter showed an increase in IgE positive cells even after 3 infections. One sheep showed twice as many IgE positive cells as the other treated sheep and the third larval infection was difficult to establish and limited in size and severity. This suggests a relationship between innate resistance to strike and the number of IgE positive cells present in skin.     

Prevalence and intensity of Thelazia spp. (Nematoda: Thelazioidea) in a Musca autumnalis (Diptera: Muscidae) population from Central Alberta

A face fly (Musca autumnalis) population near Wetaskiwin, central Alberta, Canada, was sampled 9 times from 26 July to 29 September 1988 for the early larval stages of Thelazia spp. Of 426 female flies examined, 159 (37%) contained Thelazia spp. (almost exclusively T. skrjabini), with an average worm burden of 4.2 larvae per infected fly. Prevalence ranged from 17 to 56% over 9 collections. This is the first report of Thelazia skrjabini in flies from western North America and the highest Thelazia prevalence in face flies yet reported in North America. The face fly population was also parasitized by Heterotylenchus autumnalis, with a prevalence of 5.5%.     

Investigation of the bacterial communities associated with females of Lutzomyia sand fly species from South america

Phlebotomine sand flies are vectors of Leishmania that are acquired by the female sand fly during blood feeding on an infected mammal. Leishmania parasites develop exclusively in the gut lumen during their residence in the insect before transmission to a suitable host during the next blood feed. Female phlebotomine sand flies are blood feeding insects but their life style of visiting plants as well as animals, and the propensity for larvae to feed on detritus including animal faeces means that the insect host and parasite are exposed to a range of microorganisms. Thus, the sand fly microbiota may interact with the developing Leishmania population in the gut. The aim of the study was to investigate and identify the bacterial diversity associated with wild adult female Lutzomyia sand flies from different geographical locations in the New World. The bacterial phylotypes recovered from 16S rRNA gene clone libraries obtained from wild caught adult female Lutzomyia sand flies were estimated from direct band sequencing after denaturing gradient gel electrophoresis of bacterial 16 rRNA gene fragments. These results confirm that the Lutzomyia sand flies contain a limited array of bacterial phylotypes across several divisions. Several potential plant-related bacterial sequences were detected including Erwinia sp. and putative Ralstonia sp. from two sand fly species sampled from 3 geographically separated regions in Brazil. Identification of putative human pathogens also demonstrated the potential for sand flies to act as vectors of bacterial pathogens of medical importance in addition to their role in Leishmania transmission.     

Alcohol consumption as self-medication against blood-borne parasites in the fruit fly

Plants and fungi often produce toxic secondary metabolites that limit their consumption, but herbivores and fungivores that evolve resistance gain access to these resources and can also gain protection against nonresistant predators and parasites. Given that Drosophila melanogaster fruit fly larvae consume yeasts growing on rotting fruit and have evolved resistance to fermentation products, we decided to test whether alcohol protects flies from one of their common natural parasites, endoparasitoid wasps. Here, we show that exposure to ethanol reduces wasp oviposition into fruit fly larvae. Furthermore, if infected, ethanol consumption by fruit fly larvae causes increased death of wasp larvae growing in the hemocoel and increased fly survival without need of the stereotypical antiwasp immune response. This multifaceted protection afforded to fly larvae by ethanol is significantly more effective against a generalist wasp than a wasp that specializes on D. melanogaster. Finally, fly larvae seek out ethanol-containing food when infected, indicating that they use alcohol as an antiwasp medicine. Although the high resistance of D. melanogaster may make it uniquely suited to exploit curative properties of alcohol, it is possible that alcohol consumption may have similar protective effects in other organisms.     

Hot-water immersion quarantine treatment against Mediterranean fruit fly and Oriental fruit fly (Diptera: Tephritidae) eggs and larvae in litchi and longan fruit exported from Hawaii

Immersion of litchi fruit in 49 degrees C water for 20 min followed by hydrocooling in ambient (24 +/- 4 degrees C) temperature water for 20 min was tested as a quarantine treatment against potential infestations of Mediterranean fruit fly, Ceratitis capitata (Wiedemann); and oriental fruit fly, Bactrocera dorsalis Hendel, eggs or larvae in Hawaiian litchi, Litchi chinensis Sonnerat. The 49 degrees C hot-water immersion of litchi provided probit 9 (99.9968% mortality with >95% confidence) quarantine security against eggs and first instars. There were no survivors from 15,000 each feeding and nonfeeding Mediterranean fruit fly or oriental fruit fly third instars immersed in a computer-controlled water bath that simulated the litchi seed-surface temperature profile during the 49 degrees C hot-water immersion treatment. Litchi served as the model for longan, Dimocarpus longan Lour., a closely related fruit that is smaller and also has commercial potential for Hawaii. Modified fruit infestation and holding techniques used to obtain adequate estimated treated populations from poor host fruit, such as litchi and longan, are described. Data from these experiments were used to obtain approval of a hot-water immersion quarantine treatment against fruit flies for litchi and longan exported from Hawaii to the U.S. mainland.     

Temporal occurrence of third-stage larvae of Elaeophora schneideri in Tabanus lineola hinellus on South Island, South Carolina

The occurrence of third-stage larvae of the arterial worm (Elaeophora schneideri) in populations of the horse fly, Tabanus lineola hinellus, was studied on South Island, Georgetown County, South Carolina. Over the period from mid-April to mid-October 1982, horse flies had a bimodal pattern of abundance with peak populations in June and September. During individual bimonthly horse fly collections, the prevalence of infection of T. l. hinellus ranged from 0 to 1.23%. Prevalence of infected horse flies was biphasic with peaks in mid-May (1.23%) and mid-August (1.22%), 2 to 4 wk prior to peaks in horse fly populations. The intensity of infection of T. l. hinellus increased as time progressed and was highest in late summer. This increase resulted in greater recovery of infective larvae during August-September (mean intensity = 25) than in May-June (mean intensity = 4). Both prevalence and intensity of infection of T. l. hinellus were low compared to primary intermediate hosts of E. schneideri in the western U.S.     

Recent Mediterranean fruit fly (Diptera: Tephritidae) infestations in Florida--a genetic perspective

Microsatellite and mitochondrial DNA (mtDNA) variability data were used to study infestations of the Mediterranean fruit fly, Ceratitis capitata (Wiedemann) in Florida in 1997 and 1998. A total of 132 flies collected in monitoring traps or as larvae removed from fruit were examined at three polymorphic mtDNA restriction sites and two microsatellite loci. All of the flies sampled in Florida in 1997 displayed the mitochondrial AAB haplotype and represent a novel introduction of Mediterranean fruit flies into the state. All flies collected in central Florida in 1998 also displayed the AAB haplotype. Microsatellite analysis of these specimens from 1998 detected only alleles that were present in 1997. These results strongly indicate that the 1998 Florida outbreaks were derived from the Florida populations from the previous year. According to our analyses, the Mediterranean region is the most likely source for the 1997 Florida infestation. Flies from a small outbreak in Miami Springs, Dade County, FL, early in 1998 had a different mtDNA haplotype, characterized by the AAC restriction pattern. Microsatellites of these specimens showed significant differences in their allelic distribution from AAB flies, indicating an origin from a separate source population. South America is the most likely source for the Miami Springs flies.