To identify the phlebotomine sand fly populations of the eastern coast of Tunisia, an entomological survey was carried out between September and October 2005 at 71 sites located in three districts. CDC light traps and sticky papers were used to collect a total of 2,138 phlebotomine sand flies representing nine species. The predominant species occurring on the eastern coast of Tunisia are, in order of abundance, Phlebotomus longicuspis Nitzulescu, 1930 (40%); Phlebotomus papatasi Scopoli, 1786 (21%); Sergentomyia minuta Parroti Adler & Theodor, 1927 (19%); Phlebotomus perniciosus Newstead, 1911 (9.5%); Phlebotomus chabaudi Croset, Abonnenc & Rioux, 1970 (9%); Sergentomyia fallax Parrot, 1921 (1.5%); Sergentomyia dreyfussi Parrot, 1933 (0.23%); Phlebotomus langeroni Nitzulescu, 1930 (0.05%); and Phlebotomus perfiliewi Parrot, 1930 (0.05%). Species involved in the transmission of Leishmania, namely P. papatasi and P. perniciosus, represent 31% of the total number of flies captured. In the central sites (district of Monastir), P. longicuspis predominates, P. perniciosus predominates in the northern sites (district of Sousse) and P. papatasi in the southern sites (district of Mahdia), which is consistent with the distribution of Leishmania infantum and L. major in this region. Analysis of the degree of presence (D) revealed that Phlebotomus papatasi was the most common species and showed the broadest distribution (D = 95%), followed by P. longicuspis and S. minuta parroti (D = 90%) and P. perniciosus and P. chabaudi (D = 86% and 68% respectively).

The egg of Ochlerotatus japonicus japonicus (Theobald) is described with the aid of variable pressure scanning electron micrographs. The egg is black, cigar shaped, and tapers ventrally. The length is ≈591 µm and the width is ≈172 µm. The outer chorionic cells are irregular in shape, either hexagonal or pentagonal, and decrease in size toward the anterior and posterior poles. Ventral tubercles typically range from three to six and contact the chorionic reticulum. Dorsal tubercles contain two large tubercles with small oval-shaped tubercles grouped around them. A large thread-shaped tubercle extends from this grouping either as a single tubercle or as a series of connected tubercles. The micropylar collar is low and discontinuous and is seldom complete. This description will aid researchers in the identification of this invasive vector species.

Georgia is on the southeastern margin of the native range of the brown recluse spider, Loxosceles reclusa Gertsch and Mulaik. The brown recluse is not a common Georgia spider and has limited distribution in the state. Using recent submissions, previously published records, and examination of museum specimens, we document the spider's presence in 31 (19.5%) of Georgia's 159 counties, with almost all being found in the northern portion. The spider was collected almost exclusively north of the Fall Line (a transition zone separating the Piedmont and the Coastal Plain geological provinces). Only two locations in the southern Coastal Plain province produced L. reclusa specimens; these southern finds are considered spiders that were transported outside their range. There were six finds of the non-native world tramp species, L. rufescens (Dufour), three south of the Fall Line. In conspicuous contrast, over a 5-yr period, a Georgia poison center database recorded 963 reports of brown recluse spider bites from 103 counties. These figures greatly outnumber the historic verifications of brown recluses in the state for both specimen quantity and county occurrence, indicating improbable spider involvement and the overdiagnosis of bites. In the southern half of the state, medical diagnoses of brown recluse spider bites have virtually zero probability of being correct. Bite diagnoses should be made with caution in north Georgia given the spider's spotty distribution with low frequency of occurrence.

A transition matrix model of the population dynamics of the European house dust mite, Dermatophagoides pteronyssinus (Trouessart) (Acari: Pyroglyphidae), is described. It can model continuously varying conditions by forming transition matrices by interpolation between known transition matrices constructed from experimental data. Finite carrying capacity is modeled by modifying the population distribution vector at each time-step by using a form of the Skellam model, which is derived from the assumption that in competition each successful animal gets all it requires, and the unsuccessful animals get insufficient resources for survival or reproduction. The transition matrix model does not require all mites to have the same survivorship, life-stage durations, fecundity, and so on. Life table data to drive the model is taken from two sources, one source of which requires using the mean and standard deviation of the duration of each stage to synthesize a range of duration times and a range of transition probabilities to the next stage, thus ensuring variability between mites. Where synthesized data are used, significant long-lasting oscillations in dust mite levels are modeled, which does not happen when modeling with unsynthesized data, and is unlikely to occur in the field. Under conditions normally met with in the microenvironment (bedding, base of carpet, soft furnishing) of D. pteronyssinus, finite carrying capacity is essential to prevent unbounded population growth. The model is compared with other workers' field data with fair agreement. It is argued that shortcomings in the available data rather than the model are the principal reasons for differences between field and modeled results.

Aedes albopictus is a mosquito originating from Asia, which has extended its range worldwide the last decades. It is a competent vector for several arboviruses. It was first described in La Réunion (an island of the South West part of the Indian Ocean) in 1913. Since then, it has become the dominant Aedes species and a serious threat to public health, especially during the two last arboviruses outbreaks of dengue (1977) and chikungunya (2005–2006). Despite its pest status, data on the biology of this vector are scarce, especially the population present in the Indian Ocean (IO), which has never been studied in detail. Therefore, the immature development, survival, longevity, fecundity, and gonotrophic cycles of Ae. albopictus were studied for an F₂ population of the IO. These biological parameters were studied in controlled conditions at eight constant temperatures (5, 10,15, 20, 25, 30, 35, and 40°C). The minimal threshold of immature stages development was found at 10.4°C and its optimum at 29.7°C. The shortest periods for immature development were found at 30°C, with in average of 8.8 d. The optimum intrinsic rate of growth (r) was observed between 25 and 30°C. The gonotrophic cycles were also evaluated, and the shortest cycles were found at 30°C (mean, 3.5 d). Those results are according to the field repartition of this species in La Réunion, allowing Ae. albopictus survival at a large range of temperatures.

The survival, absolute population size, gonotrophic cycle duration, and temporal and spatial abundance of Nyssomyia neivai (Pinto) were studied in a rural area endemic for American cutaneous leishmaniasis (ACL) in Conchal, Sõo Paulo State, southeastern Brazil, using mark-release-recapture techniques and by monitoring population fluctuation. The monthly abundance exhibited a unimodal pattern, with forest and domicile habitats having the highest relative abundances. A total of 1,873 males and 3,557 females were marked and released during the six experiments, of which 4.1–13.0% of males and 4.1–11.8% of females were recaptured. Daily survivorship estimated from the decline in recaptures per day was 0.681 for males and 0.667 for females. Gonotrophic cycle duration was estimated to be 4.0 d. Absolute population size was calculated using the Lincoln Index and ranged from 861 to 4,612 males and from 2,187 to 19,739 females. The low proportion of females that reach the age when they are potentially infective suggests that N. neivai has a low biological capacity to serve as a vector and that factors such as high biting rates and opportunistic feeding behavior would be needed to enable Leishmania (Viannia) braziliensis Vianna transmission. This agreed with the epidemiological pattern of ACL in southeastern Brazil that is characterized by low incidence, with isolated cases acquired principally within domiciliary habitats.

The recent resurgence of bed bugs, Cimex lectularius L. (Heteroptera: Cimicidae), has increased the demand for information about effective control tactics. Several studies have focused on determining the susceptibility of bed bug populations to insecticides. However, behavioral responses of bed bugs to insecticide residues could influence their efficacy. The behavioral responses of bed bugs to deltamethrin and chlorfenapyr, two commonly used insecticides for bed bug control in the United States, were evaluated. In two-choice tests, grouped insects and individual insects avoided resting on filter paper treated with deltamethrin. Insects did not avoid surfaces treated with chlorfenapyr. Harborages, containing feces and eggs and treated with a deltamethrin-based product, remained attractive to individuals from a strain resistant to pyrethroids. Video recordings of bed bugs indicated that insects increased activity when they contacted sublethal doses of deltamethrin. Insecticide barriers of chlorfenapyr or deltamethrin did not prevent bed bugs from reaching a warmed blood source and acquiring blood meals. We discuss the impact of these responses on bed bug control practices.

Ctenus medius Keyserling, 1891 (Araneae: Ctenidae) co-occurs in various microhabitats of the Brazilian Atlantic Forest and can be easily misidentified as the medically important spider Phoneutria nigriventer Keyserling, 1981 (Ctenidae). Despite being phylogenetically close to Phoneutria, no data are available about the toxic potential of Ctenus medius venom. Here we show that, although presenting different profile of protein composition, C. medius venom displays some of the toxic properties exhibited by P. nigriventer venom, including proteolytic, hyaluronidasic and phospholipasic activities, as well as the ability of causing hyperalgesia and edema. Moreover, C. medius venom interferes in the activation of the complement system in concentrations that P. nigriventer venom is inactive. Thus, these data show that venoms of spiders from Ctenidae family share important proinflammatory properties and suggest that the C. medius bite may have an important noxious effect in human accidents.

A floating, slow-release, granular formulation of Bacillus sphaericus (Neide) was used to control mosquito larvae in two suburban areas of two tropical cities: Ouagadougou and Bobo-Dioulasso, Burkina Faso. A circular area of 2 km², diameter 1,600 m, was treated in each city using a similar, smaller area 1 km away as an untreated control. Mosquito captures were made in houses in four concentric circles, from the periphery to the center; each circle was 50 m in width. Mosquitoes were captured in CDC light traps or from human landings. More than 95% of the mosquitoes were Culex quinquefasciatus (Say) (Diptera: Culicidae). The human landing catches provided twice as many mosquitoes as did the CDC traps/night/house. The treatments resulted in important reductions relative to the control area and to preintervention captures. The reduction was more prominent in the inner circle (up to 90%) than in the outer circle (50–70%), presumably because of the impact of immigrating mosquitoes from nontreated breeding sites around the intervention area. This effect was more pronounced for light trap catches than from human landings. The impact of treatment was also measured as the mean ratio of mosquito density in the two outer circles to that of the two inner circles. This ratio was ∼1:1 before the intervention and reached 1:0.43 during the intervention. This comparison does not depend on the assumption that, in the absence of intervention, the mosquito population development in the two areas would have been identical, but does depend on the homogeneity of the intervention area. The study showed that it is possible to organize mosquito control in a tropical, urban environment by forming and rapidly training teams of young people to carry out the mosquito control mostly using a biopesticide that can be applied without any tools except an iron bar to lift lids on some cesspits.

Pyrethroid insecticides play very important roles in the control of pediculosis, which is caused by human head louse (Pediculus humanus capitis De Geer) worldwide. The development of head louse resistance to pyrethroid pediculicides has become a global issue during the last two decades. Pyrethroid resistance is associated with amino acid substitutions in the α-subunit of the para-sodium channel gene; these substitutions are termed kdr-like mutations. The frequencies of four mutations (D11E, M815I, T929I, and L932F) in the sodium channel gene were studied in human head louse colonies collected from Japan by using SNaPshot method. A total of 630 head lice collected from 282 infested people were analyzed, and it was found that 55 lice of 19 colonies homozygously or heterozygously possessed kdr-like genes, in which the four mutations existed concomitantly. This suggested that the quadruple mutant haplotype is a common feature of the kdr genes of pandemic head lice. The frequencies of the occurrence of resistant genes in the total individuals tested and resistant gene-carrying colonies were 8.7 (55/630) and 6.7% (19/263), respectively. Because the resistant gene was detected in the colonies collected from 11 of the 22 prefectures surveyed, it is speculated that resistant head lice are already spread extensively in Japan. This was the first large-scale survey of pyrethroid resistant head lice in Japan.

Multiple assays are available to measure P450 activity in insects, including mosquitoes; however, each of these assays has drawbacks in terms of the number of mosquitoes required, specificity, sensitivity, cost, and/or time required to prepare active enzyme homogenates. In this study, a commercially available luminescent assay, P450-Glo, was modified and evaluated to measure P450 activity from the gut of a single larva after removal of the gut contents. We also compared this assay to an earlier developed fluorescent assay. After optimization of assay conditions, the P450-Glo assay held considerable promise to be used as an effective, inexpensive, high-throughput, and sensitive screening assay to measure P450 activities in single mosquitoes. Furthermore, we tested the utility of the single gut assay using the pyrethroid resistant Marin strain of Culex pipiens pipiens form molestus and the pyrethroid sensitive CQ-1 strain of Cx. pipiens quinqefasciatus. We observed on average 1.8-fold higher levels of P450 activity in the resistant mosquitoes in comparison to the sensitive mosquitoes. Additionally, consistent with our previous findings, distribution plots of P450 activity showed 33% of individual Marin mosquitoes had higher P450 activities than the highest activity displayed by a CQ-1 mosquito. The assay platform is highly flexible in terms of choice of tissue, method of preparation, isozyme specificity, and sample quantity and thus could easily be adapted to be used for other arthropod species.

Efficacy of a single dip treatment in coumaphos at 0.182% active ingredient was determined against all parasitic stages of Rhipicephalus (Boophilus) microplus (Canestrini) on infested cattle exposed to various levels of rainfall immediately after treatment. One group of calves remained untreated with no exposure to rainfall to serve as a negative control. A second group of cattle treated with coumaphos, but not exposed to rainfall, acted as a positive treated control. Three additional groups of coumaphos-treated cattle were exposed to 14.3, 28.6, and 42.9 mm of rainfall, respectively. In the coumaphos-treated group not exposed to a rainfall, overall mean control (99.2%) was greater and mean female engorgement weight (200 mg), egg mass weight (43 mg), and index of fecundity (IF; 2.90) were all less than any group exposed to rainfall. Although exposure to the lowest level of rainfall (14.3 mm) resulted in substantially greater control (83.7%) with lower mean egg mass weight (65 mg) and IF (62.26) than ticks exposed to 28.6 or 42.9 mm of rainfall, differences were seldom significant (P > 0.05). This suggested that higher levels of rainfall exposure adversely impacted coumaphos efficacy somewhat more than lower levels of rainfall exposure. Control remained >97% against larval ticks regardless of rainfall exposure level; however, against nymphs or adults, dramatic declines in control occurred as a result of exposure to any rainfall. Thus, the movement of coumaphos treated cattle exposed to any level of rainfall would pose a high risk of dispersing viable ticks into uninfested areas.

A naturally occurring sesquiterpene, isolongifolenone, derivatives of which have been used extensively as ingredients in the cosmetics industry, was discovered to effectively repel blood-feeding arthropods that are important disease vectors. We show that (—)-isolongifolenone deters the biting of the mosquitoes, Aedes aegypti (L.) and Anopheles stephensi Liston, more effectively than the widely used synthetic chemical repellent, N,N-diethyl-3-methyl benzamide (DEET), in laboratory bioassays. The compound also repelled blacklegged ticks, Ixodes scapularis Say, and lone star ticks, Amblyomma americanum (L.), as effectively as DEET. Isolongifolenone is easily synthesized from inexpensive turpentine oil feedstock. We are therefore confident that the compound has significant potential as an inexpensive and safe repellent for protection of large human populations against blood-feeding arthropods.

Azadirachta indica A. Juss (the neem tree), a source of limonoid insect growth regulatory (IGRs), grows well in many places in sub-Saharan Africa. We explored the potential of neem wood and bark chippings in malaria vector control by evaluating their aqueous extracts as a larvicide and growth disruptor of Anopheles gambiae s.s. (Diptera: Culicidae) under laboratory conditions. Immature stages of the mosquito were tested using WHO guidelines. Fifty percent inhibition of adult emergence (IE₅₀) of all larval instars was obtained with <0.4 g of neem chippings in 1 liter of distilled water. For pupae, significant mortality occurred at 5 g/liter. Inhibition of pupation was seen with some larvae staying as LIVs for 9 d before dying. In addition to growth retardation, reduced reaction by larvae to visual and mechanical stimuli observed at higher neem concentrations may make them more susceptible to natural predators. There were no significant differences in the sex ratio of emerged adults or wing length of females compared with the controls. High-performance liquid chromatography of aqueous extracts showed a series of constituents of varying polarity, including the limonoids nimbin and salannin, which were quantified. Azadirachtin was not detected and the observed activities are attributed to other constituents of the chippings. Such larvicides can be particularly effective where larval habitats are relatively large and readily identifiable. Aqueous extracts of neem wood chippings can be produced locally and their use has the potential to be a low-tech component of integrated malaria vector control schemes in sub-Saharan Africa.

Larval and nymphal Ixodes pacificus Cooley and Kohls, I. (Ixodes) jellisoni Cooley and Kohls, and Dermacentor occidentalis Marx were tested for host preference when simultaneously presented with a deer mouse (Peromyscus maniculatus Wagner), California kangaroo rat (Dipodomys californicus Merriam), western fence lizard (Sceloporus occidentalis Baird and Girard), and California towhee (Pipilo crissalis Vigors) in an experimental apparatus. Differences were observed in the preferences among the three species and between life stages. More larvae of all species approached and contacted hosts than did nymphs. Subadult I. pacificus entered all host-containing chambers in the highest numbers and remained on lizards most often after contact. Subadult I. jellisoni entered and remained in the chambers containing kangaroo rats, while rejecting mice, lizards, and birds as hosts. Subadult D. occidentalis most frequently entered rodent-containing chambers and contacted these hosts. After overnight exposure to all nonavian hosts, only I. pacificus parasitized and fed successfully on all three animals. I. jellisoni fed only on kangaroo rats and D. occidentalis fed only on rodents. Molting success ranged from ≈66 to 95% among tick species and stages. We concluded that, under laboratory conditions, I. pacificus larvae and nymphs prefer western fence lizards, but also will parasitize rodents. Dermacentor occidentalis immatures use deer mice and kangaroo rats similarly, whereas I. jellisoni subadults exclusively parasitize kangaroo rats. California towhees are considerably less attractive as hosts for these three ticks. These host preferences are consistent with what is known about the natural feeding habits of all three ticks.

To gain insight into the transmissibility of bacteria by house flies, the temporal and spatial fate of green fluorescent protein (GFP)-expressing motile and nonmotile strains of Aeromonas hydrophila was examined within the alimentary canal. Liquid food consumed by house flies is first stored in the crop and then is regurgitated and/or passed into the midgut. Once within the midgut, food is contained inside a double-layered peritrophic matrix (PM), with the inner layer enveloping digested material and forming fecal pellets for excretion. Between 1 and 12 h after ingestion, and irrespective of motility, live GFP A. hydrophila adhered to the luminal surfaces of the crop and inner PM of bacteria-fed flies. However, some nonadherent, motile bacteria moved freely within the PM lumen in the anteromedial midgut, whereas fecal pellets (lysed bacteria) continued passing posteriorly. At 12–24 h, adhered bacteria of both strains were lysed in the distal midgut, compressed into fecal pellets, and excreted. Viable bacteria in the crop visually exceeded numbers within these fecal pellets. Culture recovery at the same time points showed a 1,000-fold increase of viable bacteria at 2 h, presumably in the crop, with a temporal decline thereafter. Further, viable bacteria were recovered from vomit specks and orally contaminated substrates up to 2 h after feeding but never from feces. These results suggest that A. hydrophila is a transient resident of the house fly alimentary canal and is only orally transmissible for a short time after ingestion. Thus, regurgitation may be more significant than fecal transmission in the spread of some house fly transmitted bacterial diseases.

Lyme disease in the United States is caused by the bacterial spirochete Borrelia burgdorferi s.s. (Johnson, Schmid, Hyde, Steigerwalt, and Brenner), which is transmitted by tick vectors Ixodes scapularis (Say) and I. pacificus (Cooley and Kohls). Borrelia lonestari, transmitted by the tick Amblyomma americanum L., may be associated with a related syndrome, southern tick-associated rash illness (STARI). Borrelia lonestari sequences, reported primarily in the southeastern states, have also been detected in ticks in northern states. It has been suggested that migratory birds may have a role in the spread of Lyme disease spirochetes. This study evaluated both migratory waterfowl and nonmigratory wild turkeys (Meleagris gallopavo silvestris, Eastern wild turkey) for B. burgdorferi and B. lonestari DNA sequences. A total of 389 avian blood samples (163 migratory birds representing six species, 125 wild turkeys harvested in habitats shared with migratory birds, 101 wild turkeys residing more distant from migratory flyways) were extracted, amplified, and probed to determine Borrelia presence and species identity. Ninety-one samples were positive for Borrelia spp. Among migratory birds and turkeys collected near migration routes, B. burgdorferi predominated. Among turkeys residing further away from flyways, detection of B. lonestari was more common. All A. americanum ticks collected from these areas were negative for Borrelia DNA; no I. scapularis were found. To our knowledge, this represents the first documentation of B. lonestari among any birds.

West Nile virus (WNV) has remained epidemic in Kern County, CA, since its introduction in 2004 through 2007 when the human case annual incidence increased from 6–8 to 17 per 100,000, respectively. The 2007 increase in human infection was associated with contradicting surveillance indicators, including severe drought, warm spring but cool summer temperature anomalies, decreased rural and urban mosquito abundance but increased early season infection in urban Culex quinquefasciatus Say, moderate avian “herd immunity,” and declines in the catch of competent (western scrub-jay and house finch) and noncompetent (California quail and mourning dove) avian species. The decline in these noncompetent avian hosts may have increased contact with competent avian hosts and perhaps humans. The marked increase in home foreclosures and associated neglected swimming pools increased urban mosquito production sites, most likely contributing to the urban mosquito population and the WNV outbreak within Bakersfield. Coalescing five surveillance indicators into a risk assessment score measured each half month provided 2- to 6-wk early warning for emergency planning and was followed consistently by the onset of human cases after reaching epidemic conditions. St. Louis encephalitis virus (SLEV) antibody was detected rarely in wild birds but not mosquitoes or sentinel chickens, indicating that previously infected birds were detected in Kern County, but SLEV reintroduction was not successful. In contrast, western equine encephalitis virus (WEEV) was detected during 3 of 5 yr in Culex tarsalis Coquillett, sentinel chickens, and wild birds, but failed to amplify to levels where tangential transmission was detected in Aedes mosquitoes or humans. A comparison of transmission patterns in Kern County to Coachella Valley in the southeastern desert of California showed the importance of mosquito phenology and spatial distribution, corvids, or other avian “super spreaders” and anthropogenic factors in WNV epidemiology.

We developed a single-step real-time fluorescence resonance energy transfer (FRET) multiplex polymerase chain reaction (PCR) merged with melting curve analysis for the detection of Wuchereria bancrofti and Brugia malayi DNA in blood-fed mosquitoes. Real-time FRET multiplex PCR is based on fluorescence melting curve analysis of a hybrid of amplicons generated from two families of repeated DNA elements: the 188 bp SspI repeated sequence, specific to W. bancrofti, and the 153-bp HhaI repeated sequence, specific to the genus Brugia and two pairs of specific fluorophore-labeled probes. Both W. bancrofti and B. malayi can be differentially detected in infected vectors by this process through their different fluorescence channel and melting temperatures. The assay could distinguish both human filarial DNAs in infected vectors from the DNAs of Dirofilaria immitis- and Plasmodium falciparum-infected human red blood cells and noninfected mosquitoes and human leukocytes. The technique showed 100% sensitivity and specificity and offers a rapid and reliable procedure for differentially identifying lymphatic filariasis. The introduced real-time FRET multiplex PCR can reduce labor time and reagent costs and is not prone to carry over contamination. The test can be used to screen mosquito vectors in endemic areas and therefore should be a useful diagnostic tool for the evaluation of infection rate of the mosquito populations and for xenomonitoring in the community after eradication programs such as the Global Program to Eliminate Lymphatic Filariasis.

A three-dimensional (3-D) model of the digestive system of Periplaneta americana was built for the first time based on hematoxylin and eosin (H&E) staining, the study of multiple cross-sections of the larval cockroach, and 3-D reconstruction technology. The digestive system of P. americana includes the foregut, midgut, and hindgut and takes up most of the celom. The foregut comprises almost one half of the digestive system (43.57%). The midgut, the critical region for digestion and absorption, has the second highest volume ratio (35.21%). The hindgut, with the lowest volume ratio (21.22%), includes the ileum, colon, and rectum. After the ileal valve is the colon. The 3-D model presented in this paper provides a stereoscopic view for studying the adjacent relationship and arrangement of different gut sections of P. americana.

The male sex determining factor (M) in the house fly is linked to the Y chromosome in the ancestral condition, but can also be linked to another chromosome (I-V or X). However, descriptions of the linkage and frequency of M factors in different populations throughout the world are vastly incomplete. We collected house flies from a dairy in Ipswich, Australia, and determined that M was linked to chromosomes II, III, IV, and Y. Most males (69.8%) were homozygous for M on autosome II and/or III, and 92.3% of the males had multiple M factors. In all, there were 13 different male genotypes found. The high frequency of M, the presence of M on four different linkage groups, and the large number of male genotypes found in this population make it unique relative to other populations of house flies that have been examined.