Robert K. Velten

An automated feeding apparatus for in vitro maintenance of the human head louse, Pediculus capitis (Anoplura: Pediculidae).

Abstract An automated feeding apparatus was developed to maintain the human head louse (Pediculus capitis DeGeer) in vitro. With the use of valves and timers, banked human blood and saline from refrigerated reservoirs were pumped into and flushed out of the system every 7 d. During this rotational interval, bloodmeals were provided to head lice continuously and ad libitum through a stretched Nescofilm-silicone sandwich membrane. Compared with our previous in vitro human head louse-rearing apparatus, greater numbers of lice could be fed simultaneously with minimal human monitoring. Development of second to third instars and third instars to adults was significantly faster when lice were reared in vivo than on either of the in vitro rearing systems; there was no significant difference in the duration of the first instar. Although fecundity and hatch rates were significantly higher for female lice reared in vivo, similar trends have been observed for other membrane-fed arthropods. Body lice (Pediculus humanus L.) and bed bugs (Cimex lectularius [L.]) also completed most of their life cycle on this apparatus. Our automated mass-rearing system has broad applications for maintaining fluid-sucking ectoparasites and will facilitate various toxicological, behavioral, and disease-transmission investigations.

Failure of a Permethrin Treatment Regime to Protect Cattle Against Bluetongue Virus

Abstract Holstein heifers in a confined feedlot setting on a southern California dairy were either sprayed individually along the ventral midline using 0.2% permethrin (250 ml/animal) (two pens) or were not treated (two pens). Treatments (n = 6 dates) were applied every 2 wk during the peak fall bluetongue virus transmission season (22 August–29 October). Animals seronegative for bluetongue virus antibodies at the initial bleeding on 15–18 September (n = 106 in the treatment pens and n = 117 in the control pens) were bled again for testing 2 mo later (12–13 November). Seroconversion rates were not significantly different: 56% for the treated animals and 48% for the controls (P > 0.2). The area has many essentially contiguous, confinement dairies with wastewater ponds that produce large numbers of Culicoides sonorensis Wirth & Jones, the primary bluetongue virus vector. Further, these dairies presumably provided a large reservoir of virus-infected cattle to infect vectors in the immediate area. Under these severe virus challenge conditions, permethrin applied at 2-wk intervals failed to reduce exposure to bluetongue virus.

Feeding and survival of Culicoides sonorensis on cattle treated with permethrin or pirimiphos-methyl.

The persistence of permethrin (5% a.i.) and pirimiphos‐methyl (27% a.i.), applied to the dorsum of calves in the field against Culicoides sonorensis Wirth and Jones (Diptera: Ceratopogonidae), was estimated using a hair‐blood‐feeding bioassay in the laboratory. Hair clippings were taken before treatment and 3, 7, 14, 21, 28, 42 and 56 days after treatment from the dorsum, side and belly of treated and control calves. Laboratory‐reared insects were allowed to feed through thin hair layers and a parafilm membrane on sheep blood warmed using a water‐jacketed feeder. Some intoxication after exposure to hair was noted up to 28 days after treatment with permethrin and up to 14 days after treatment with pirimiphos‐methyl. Hair from the dorsum caused more intoxication for a longer period than hair from other body regions. Permethrin and pirimiphos‐methyl applied to the back did not significantly reduce overall engorgement (body regions pooled) after treatment. Permethrin residues on hair remained far higher on the back than other body regions and were related to insect intoxication and reduction in engorgement in the laboratory. Residues on belly hair never exceeded 12 p.p.m. and did not result in significantly reduced feeding at any time. Engorged insects that exhibited sublethal intoxication from feeding through permethrin‐treated hair did recover and matured numbers of eggs comparable to controls. Field trials using treated and control calves and enclosure nets showed that dorsal applications of 5% permethrin were not effective in reducing engorgement, despite some intoxication. Vacuum samples from a calf showed that C. sonorensis fed primarily on the belly. A 0.2% permethrin application on the belly (250 ml) did result in > 80% reduction of C. sonorensis in the enclosure nets at 3 and 7 days after treatment, but activity had subsided by 10 days after treatment. The utility of insecticidal treatments for suppression of this vector is discussed.

Two Pathogens and One Disease: Detection and Identification of Flea-Borne Rickettsiae in Areas Endemic for Murine Typhus in California

ABSTRACT Results of an environmental assessment conducted in a newly emergent focus of murine typhus in southern California are described. Opossums, Didelphis virginiana Kerr, infested with cat fleas, Ctenocephalides felis Buché, in the suburban area were abundant. Animal and flea specimens were tested for the DNA of two flea-borne rickettsiae, Rickettsia typhi and Rickettsia felis. R. felis was commonly detected in fleas collected throughout this area while R. typhi was found at a much lower prevalence in the vicinity of just 7 of 14 case-patient homes identified. DNA of R. felis, but not R. typhi, was detected in renal, hepatic, and pulmonary tissues of opossums. In contrast, there were no hematologic polymerase chain reaction findings of R. felis or R. typhi in opossums, rats, and cats within the endemic area studied. Our data suggest a significant probability of human exposure to R. felis in the area studied; however, disease caused by this agent is not recognized by the medical community and may be misdiagnosed as murine typhus using nondiscriminatory serologic methods.

Variation in the NS3/NS3A gene of bluetongue viruses contained in Culicoides sonorensis collected from a single site in southern California

To determine the variability of the NS3/NS3A gene of field strains of BTV contained in Culicoides sonorensis collected from a single site in California (CA), the NS3/NS3A gene was directly amplified and sequenced from 22 pools of C. sonorensis and compared with those of previously characterized field isolates from CA, as well as to viruses that caused recent outbreaks of bluetongue disease in ruminants in CA. Phylogenetic analysis established that the NS3/NS3A gene of strains of BTV contained in C. sonorensis collected from the site exists as a heterogeneous population. The two most divergent nucleotide sequences of the NS3/NS3A genes of these viruses differed by 2.5% (18 nucleotides). Comparison with the NS3/NS3A gene sequences from viruses that caused recent instances of bluetongue disease in ruminants in CA indicated that BTV strains from different geographic regions can exhibit a higher degree of genetic heterogeneity (up to 6.6%; 0-48 nucleotide differences) than those contained in C. sonorensis collected from a single site.

Host (Avian) Biting Preference of Southern California Culex Mosquitoes (Diptera: Culicidae)

ABSTRACT The host preference of a vector mosquito species plays a significant role in determining human and animal risk of infection with mosquito-transmitted pathogens. Host preferences of common southern California Culex species for four bird species, American crow (Corvus brachyrhynchos), house sparrow (Passer domesticus), house finch (Carpodacus mexicanus), and mourning dove (Zenaida macroura), were examined by determining the proportion of each mosquito species that successfully engorged on each of the four bird species presented equally within a net trap to wild host-seeking mosquitoes. Bloodmeals in engorged mosquitoes captured within the net trap were identified to avian species by using a multiplex polymerase chain reaction assay targeting the cytochrome b gene sequence. There were significant differences in host selection by all three Culex species captured in numbers sufficient for analysis, with Culex erythrothorax Dyar preferentially biting American crows, Culex tarsalis Coquillett preferentially biting house sparrows, and Culex quinquefasciatus Say preferentially biting house finches. All three Culex species demonstrated more frequent engorgement on passerine birds (sparrows, finches, and crows) than the nonpasserine mourning dove. A greater preference for passerine birds might be expected to increase the transmission of pathogens, such as West Nile virus, to which passerine birds are particularly competent hosts.