Persistence and Retention of Porcine Reproductive and Respiratory Syndrome Virus in Stable Flies (Diptera: Muscidae).

We investigated the acquisition of porcine reproductive and respiratory syndrome (PRRS) virus by the stable fly (Diptera: Muscidae; Stomoxys calcitrans (L.)) through a bloodmeal, and virus persistence in the digestive organs of the fly using virus isolation and quantitative reverse-transcription PCR (qRT-PCR). Stable flies were fed blood containing live virus, modified live vaccine virus, chemically inactivated virus, or no virus. Stable flies acquired PRRSV from the bloodmeal and the amount of virus in the flies declined with time, indicating virus did not replicate in fly digestive tissues. Virus RNA was recovered from the flies fed live virus up to 24 h postfeeding using virus isolation techniques and 96 h using qRT-PCR. We further examined the fate of PRRSV in the hemolymph of the flies following intrathoracic injection to bypass the midgut barrier. PRRSV was detected in intrathoracically inoculated adult stable flies for 10 d using qRT-PCR. In contrast to what we observed in the digestive tract, detectable virus quantities in the intrathoracically inoculated stable flies followed an exponential decay curve. The amount of virus decreased fourfold in the first 3 d and remained stable thereafter, up to 10 d.

Dispersion and sampling of adult Dermacentor andersoni in rangeland in Western North America.

A fixed precision sampling plan was developed for off-host populations of adult Rocky Mountain wood tick, Dermacentor andersoni (Stiles) based on data collected by dragging at 13 locations in Alberta, Canada; Washington; and Oregon. In total, 222 site-date combinations were sampled. Each site-date combination was considered a sample, and each sample ranged in size from 86 to 250 10 m2 quadrats. Analysis of simulated quadrats ranging in size from 10 to 50 m2 indicated that the most precise sample unit was the 10 m2 quadrat. Samples taken when abundance < 0.04 ticks per 10 m2 were more likely to not depart significantly from statistical randomness than samples taken when abundance was greater. Data were grouped into ten abundance classes and assessed for fit to the Poisson and negative binomial distributions. The Poisson distribution fit only data in abundance classes < 0.02 ticks per 10 m2, while the negative binomial distribution fit data from all abundance classes. A negative binomial distribution with common k = 0.3742 fit data in eight of the 10 abundance classes. Both the Taylor and Iwao mean-variance relationships were fit and used to predict sample sizes for a fixed level of precision. Sample sizes predicted using the Taylor model tended to underestimate actual sample sizes, while sample sizes estimated using the Iwao model tended to overestimate actual sample sizes. Using a negative binomial with common k provided estimates of required sample sizes closest to empirically calculated sample sizes.

Assessment of Stomoxys calcitrans (Diptera: Muscidae) as a vector of porcine reproductive and respiratory syndrome virus.

Porcine Reproductive and respiratory syndrome (PRRS) is a globally significant swine disease caused by an arterivirus. The virus replicates in alveolar macrophages of infected pigs, resulting in pneumonia in growing pigs and late-term abortions in sows. Outbreaks occur on disparate farms within an area despite biosecurity measures, suggesting mechanical transport by arthropods. We investigated the vector potential of stable flies, Stomoxys calcitrans (L.) (Diptera: Muscidae), in the transmission of porcine reproductive and respiratory syndrome virus (family Arteriviridae, genus Arterivirus, PRRSV) under laboratory conditions. Stable flies were collected around PRRS-negative boar stud barns in North Carolina and tested for presence of the virus. Stable flies were collected on alsynite traps placed near the exhaust fan of the close-sided tunnel-ventilated buildings, suggesting blood seeking flies are attracted by olfactory cues. No flies were positive for PRRSV. We assessed transmission of the virus through an infective bite by feeding laboratory reared stable flies on blood containing virus and transferring them to naive pigs for subsequent bloodmeals. Transmission of the virus to naive pigs by infective bites failed in all attempts. The volume of blood contained within the closed mouthparts of the stable fly seems to be insufficient to deliver an infective dose of the virus. Stable flies are unlikely to transmit PRRSV from one pig to another while blood feeding. The fate of the virus after a bloodmeal remains to be determined.

Activity of Bacillus thuringiensis isolates against immature horn fly and stable fly (Diptera: Muscidae).

We screened 85 isolates of Bacillus thuringiensis (Berliner), making up 57 different subspecies, and two isolates of Bacillus sphaericus (Meyer and Neide) for activity against immature horn flies, Haematobia irritans (L.), and stable flies, Stomoxys calcitrans (L.). The majority of B. thuringiensis and the B. sphaericus isolates had little or no activity against horn fly and stable fly. Approximately 87% of the isolates caused < 50% mortality of horn fly larvae and 64% caused < 25% mortality. For stable fly, 95% of the isolates caused < 50% mortality, and 93% caused < 25% mortality. Five isolates were highly toxic to horn fly and stable fly immatures. These isolates were B. t. tolworthi 4L3, B. t. darmstadiensis 4M1, B. t. thompsoni 401, B. t. thuringiensis HD2, and B. t. kurstaki HD945. The LD50 values ranged from 2.2 to 7.9 x 10(6) spores per g manure for horn fly and from 6.3 to 35 x 10(6) spores per g media for stable fly. These were consistently more toxic compared with the B. t. israelensis isolates examined. All had DNA that hybridized with cry1Aa, cry1Ab, and cry1Ac toxin probes, three hybridized with a cry1B probe, and two hybridized with a cry2A probe. These may have potential for use in integrated management of pest flies.

Retention of Escherichia coli by house fly and stable fly (Diptera: Muscidae) during pupal metamorphosis and eclosion.

Populations of Escherichia coli obtained by feeding larval house flies, Musca domestica L. and stable flies, Stomoxys calcitrans (L.), persisted through the pupal stage. The abundance of E. coli in house fly pupae increased initially then declined before adult emergence. Abundance of E. coli in stable fly pupae increased through pupal development and remained high. Infected stable fly pupal cases typically contained more E. coli than house fly pupal cases. A greater proportion of emerging adult house flies were infected with E. coli compared with stable flies; however, the abundance of E. coli on infected flies was similar between species. Adult flies contained 0.04-0.19% of the E. coli in the pupal cases. The proportion of infected house fly adults and the amount of E. coli on the infected flies were related to the levels of E. coli in the pupal cases; however, these relationships did not occur with the stable fly. Results suggest that retention of E. coli from larval to adult house flies could play a role in the transmission and spread of E. coli, whereas stable fly adults probably play a minor role in E. coli spread. However, pupae of both species have potential to act as reservoirs for E. coli.

Persistence of Escherichia coli in immature house fly and stable fly (Diptera: Muscidae) in relation to larval growth and survival.

The persistence of Escherichia coli in artificially fed larvae was examined for up to 48 h after ingestion by house flies, Musca domestica L., and stable flies, Stomoxys calcitrans (L.). The rate of change in the E. coli load was similar for both species for up to 5 h after ingestion. Up to 48 h after ingestion, abundance of E. coli declined in immature house flies but remained constant in immature stable flies. When different E. coli concentrations were fed to larvae, the abundance of E. coli increased in stable fly larvae regardless of the initial concentration. The E. coli load in house fly larvae increased when larvae were fed a low concentration of bacteria, but it declined when larvae were fed a high concentration of bacteria. Survival of house fly and stable fly larvae averaged 62 and 25%, respectively, when reared on pure E. coli cultures. These observations suggest that house fly larvae digest E. coli and use it as a food source but stable fly larvae do not.