To date, there has been no confirmed, indigenously acquired case of arthropod-borne viral disease in New Zealand, but this may change in the near future due to the presence of exotic vectors and regular influx of infected humans. The risk of a disease outbreak may be aggravated if other exotic mosquito vectors become established, in particular Aedes albopictus (Skuse), a species that has already been intercepted several times in New Zealand. In this study, the possible means of invasion and dispersal of Ae. albopictus in northern New Zealand are discussed, along with the factors that should facilitate its establishment in the event it evades border controls.

The southern cattle tick, Boophilus microplus (Canestrini), causes annual economic losses in the hundreds of millions of dollars to cattle producers throughout the world, and ranks as the most economically important tick from a global perspective. Control failures attributable to the development of pesticide resistance have become commonplace, and novel control technologies are needed. The availability of the genome sequence will facilitate the development of these new technologies, and we are proposing sequencing to a 4–6X draft coverage. Many existing biological resources are available to facilitate a genome sequencing project, including several inbred laboratory tick strains, a database of ≈45,000 expressed sequence tags compiled into a B. microplus Gene Index, a bacterial artificial chromosome (BAC) library, an established B. microplus cell line, and genomic DNA suitable for library synthesis. Collaborative projects are underway to map BACs and cDNAs to specific chromosomes and to sequence selected BAC clones. When completed, the genome sequences from the cow, B. microplus, and the B. microplus-borne pathogens Babesia bovis and Anaplasma marginale will enhance studies of host–vector–pathogen systems. Genes involved in the regeneration of amputated tick limbs and transitions through developmental stages are largely unknown. Studies of these and other interesting biological questions will be advanced by tick genome sequence data. Comparative genomics offers the prospect of new insight into many, perhaps all, aspects of the biology of ticks and the pathogens they transmit to farm animals and people. The B. microplus genome sequence will fill a major gap in comparative genomics: a sequence from the Metastriata lineage of ticks. The purpose of the article is to synergize interest in and provide rationales for sequencing the genome of B. microplus and for publicizing currently available genomic resources for this tick.

Anaplasma marginale Theiler is a tick-borne pathogen that causes anaplasmosis in cattle. There are ≈20 tick species worldwide that are implicated as vectors of this pathogen. In the United States, Dermacentor andersoni Stiles and Dermacentor variabilis (Say) are the principal vectors. The risk of transmission of anaplasmosis to cattle has been largely based on the distribution of D. andersoni in the United States. We developed a centralized geographic database that incorporates collection records for D. andersoni from two large national databases. We reviewed the geographic records in each database and postings from MEDLINE and AGRICOLA to produce a national county-level distribution map based on a total of 5,898 records. The records spanned the period from 1903 through 2001 with the majority between 1921 to 1940. Populations of D. andersoni were recorded from 267 counties in 14 states and were distinguished as either established or reported. We found 180 counties with established populations of D. andersoni and 87 counties with reported occurrences in 14 states with the majority of established populations reported from Montana, Idaho, and Oregon. D. andersoni populations in the United States currently extend from the western portions of Nebraska and the Dakotas westward to the Cascade Mountains and from the northern counties of Arizona and New Mexico northward to the Canadian border. The data will be useful for identifying regions at increased risk of acquiring anaplasmosis in the United States. Based upon the database collection records, we also present a summary of recorded hosts for D. andersoni and comments on its seasonal occurrence.

Growth characteristics and whole body carbon (C), nitrogen (N), and phosphorus (P) concentrations were examined for the southern house mosquito, Culex quinquefasciatus Say, and Culex tarsalis Coquillett, reared on chemostat-grown bacteria, Pseudomonas aeruginosa. Whole body percentage of C, N, and P of Cx. quinquefasciatus larvae did not differ significantly across three bacterial concentrations (1, 5, and 10 mg of dry mass/liter) and two bacterial quality treatments (culture medium containing 5 μM P versus 50 μM P); whereas the P content of Cx. tarsalis larvae differed between the bacterial quality treatments. Low concentrations of high or low P bacteria decreased mass-specific growth rate (MGR), whereas intermediate and high bacterial concentrations affected MGR asymmetrically, depending on species. High concentrations of P-rich bacteria enhanced the growth rates of Cx. quinquefasciatus larvae relative to growth on the low P diets. Cx. tarsalis larvae reared on low P bacteria grew ≈3- to 4 times faster than larvae reared on high P bacteria. The observed asymmetric response in MGR may have been because of differential tolerance in larvae to putative toxins present in P. aeruginosa and may provide one reason why Cx. tarsalis larvae are not found in hypereutrophic aquatic habitats.

A series of behavioral tests with Aedes aegypti (L.), Anopheles stephensi Liston, mosquitoes, and the sand fly Phlebotomus papatasi Scopoli in the presence of Deet, SS220, and Picaridin topically applied to the skin of human volunteers showed that the insects were deterred from feeding on and repelled from surfaces emanating the compounds. When offered a 12- or 24-cm² area of skin, one-half treated with compound and one-half untreated, the insects fed almost exclusively on untreated skin. The sand flies and mosquitoes did not at any time physically contact chemically treated surfaces. When treated and untreated skin areas were covered with cloth, insects contacted, landed, and bit only through cloth covering untreated skin. These observations provided evidence that the compounds deterred feeding and repelled insects from treated surfaces primarily as a result of olfactory sensing. When cloth, one-half untreated and one-half treated with chemical, was placed over untreated skin, insects only touched and specifically bit through the untreated cloth. This showed that the activity of the chemicals does not involve a chemical × skin interaction. In the presence of any of the three chemicals, no matter how they were presented to the insects, overall population biting activity was reduced by about one-half relative to controls. This reduction showed a true repellent effect for the compounds. Results clearly showed that Deet, SS220, and Picaridin exert repellent and deterrent effects upon the behavior of mosquitoes and sand flies. Heretofore, the combined behavioral effects of these compounds upon mosquito and sand fly behavior were unknown. Moreover, protection afforded by Deet, SS220, and Picaridin against the feeding of these three disease vectors on humans is mechanistically a consequence of the two chemical effects.

The presence of Aedes albopictus (Skuse) was recently confirmed for the first time in northwestern Greece. This location is within the distribution range of a morphologically similar species, Aedes cretinus Edwards, and is a potentially favorable region for the reintroduction of Aedes aegypti (L.). It was thus compelling to use methods in addition to morphology-based keys to correctly identify specimens badly damaged, rubbed, or otherwise altered in their external characteristics. It was decided to use molecular techniques as a novel and reliable method for differentiating the three Stegomyia species. The nuclear internal transcribed spacer 2 (ITS2) fragments from morphologically identified Ae. albopictus and Ae. cretinus specimens were amplified, and their sequences were compared with those in GenBank for Ae. albopictus, Ae. cretinus, and Ae. aegypti. Also, mitochondrial cytochrome oxidase I (COI) fragments were amplified for Ae. albopictus and Ae. cretinus (so far not available in GenBank) and compared with Ae. aegypti fragments. ITS2 and COI sequences generated in our study were deposited in GenBank and could be useful in future studies of mosquitoes by other research workers.

Insecticides are one of the major tools for controlling vector populations and for reducing the transmission of human pathogens. However, there are few new insecticides being developed and marketed for vector control. Herein, we report on the toxicity of six novel insecticides to both adult and larval Aedes aegypti (L). and the toxicity of three novel insect growth regulators (IGRs) to larvae. Four insecticides were highly or moderately toxic to larvae with LC₅₀ values of 16 (chlorfenapyr), 70 (hydramethylnon), 79 (indoxacarb), and 84 ng/ml (imidacloprid). Diafenthiuron and chlorfenapyr were moderately toxic to adult mosquitoes with LC₅₀ values of 13 and 92 ng/cm², respectively. Imidacloprid was strongly synergized by piperonyl butoxide (PBO) in Ae. aegypti adults, suggesting that neonicotinoids are intrinsically very toxic to adult mosquitoes (in the absence of detoxification). The effect of PBO on the toxicity in adults and larvae was considerably different, both in terms of the insecticides that were synergized (or antagonized for chlorfenapyr versus adults) and in terms of the degree of synergism. This result implies that the cytochrome P450s involved in metabolism of these insecticides are different between adults and larvae. Pyriproxyfen was confirmed as a potent IGR (EC₅₀ of 0.0017 ng/ml) for mosquitoes, although tebufenozide lacked activity. The potential for use of these materials in mosquito control is discussed.

The acquisition of Borrelia burgdorferi by the larvae of competent and refractory ixodid ticks was assessed by quantitative polymerase chain reaction (PCR). Larvae were fed on infected mice, and the spirochete loads were determined during feeding and up to 93 d postfeeding. Amblyomma americanum (L.) was refractory to B. burgdorferi infection, with almost no detection of spirochete DNA during or postfeeding. In contrast, Ixodes scapularis Say supported high loads of spirochetes (10³–10⁴ per larva). In Dermacentor variabilis (Say), B. burgdorferi uptake was reduced, with an average of 16 spirochetes per larvae acquired after 4 d of feeding, representing 1/195 of the counts in I. scapularis. However, during the first day postfeeding, the spirochete growth rate in D. variabilis reached 0.076 generations per hour, 7.7 times greater than the highest growth rate detected in I. scapularis. D. variabilis supported intense spirochete growth up to the fourth day postinfection, when the counts increased to an average of 282 spirochetes per larvae or 1/8.5 of the I. scapularis counts 4 d postfeeding. The kinetics of spirochete growth was unstable in D. variabilis compared with I. scapularis, and transmission of B. burgdorferi by D. variabilis could not be demonstrated. A cofeeding experiment indicated that I. scapularis feeding increased A. americanum spirochete uptake. These collective results indicate suboptimal conditions for B. burgdorferi uptake and colonization within A. americanum or the presence of anti-Borrelia factor(s) in this nonpermissive tick species.

Egg hatching has been studied in Aedes aegypti (L.) through scanning electron microscopy. The first sign of egg hatching is a small protrusion on the eggshell in the anterior pole. The larval movement provokes a crack in the eggshell with the egg buster located in the dorsal head. The egg buster provokes a small transverse fissure in the eggshell that gradually increases in the chorion. Then, the rupture is completed around the eggshell. The separation of the anterior pole occurs, showing the dorsal region of the larva head with the egg buster and the cap. After sequential movements, the larva looses the cap. Finally, the first instar is ready to be free showing details of its body with the egg buster over its head. This structure is a cuticular formation, similar to a cone structure that ends in a very fine tip and emerges from a pear-like depression with high rounded borders. Our results describe the anatomy of the egg hatching process in Ae. aegypti, showing details of the participation of the egg buster.

Oral susceptibility to infection with bluetongue virus (family Resviridae, genus Orbivirus, BTV) serotype 9 was characterized in three Palaearctic species of Culicoides (Diptera: Ceratopogonidae). Variation in susceptibility to infection by using a recently described feeding technique was shown to occur between populations of Culicoides obsoletus Meigen complex midges from different geographic regions of the United Kingdom with virus infection rates varying from 0.4 to 7.4% of those tested. Susceptibility to infection was consistent on an annual basis at selected sites. Prevalence of infection in the most susceptible populations of both the C. obsoletus and Culicoides pulicaris L. complexes was comparable with that of Culicoides imicola Kieffer, the major vector of BTV in southern Europe and throughout Africa, when using the same feeding method and virus. These results are discussed with reference to the potential threat of the virus to susceptible livestock in northern Europe.

Malaria in Sri Lanka is closely associated with the river systems in the country. Construction of major dams across rivers has resulted in year-round pooling and anopheline mosquito breeding in river beds below the dams. This study was carried out to determine the larvivorous activity of fish found in river bed pools below the dams to identify the potential fish species for anopheline larval control in these areas. Fish species in five river beds below the dams, namely, Laxapana, Kotmale, Nilambe, Victoria, and Rantembe, were identified in June 2000. The larvivorous potential of the 12 fish species collected was determined in the laboratory based on the number of larvae consumed per fish within 10 min and an hour. Danio malabaricus Jerdon, Oreochromis mossambicus Peters, Oreochromis niloticus L., and Poecilia reticulata Peters consumed nine or more larvae per fish within 10 min. Aplocheilus dayi Steindachner and Rasbora daniconius Hamilton consumed nine and eight larvae, respectively, within 1 h. O. mossambicus, O. niloticus, and P. reticulata consumed 10 larvae per fish within 10 min. Despite some species having a high larvivorous potential in the laboratory, field studies are required to determine the feasibility of using these fish for anopheline mosquito control in the river bed pools below the dams.

Host-seeking mosquitoes were sampled in bird-baited traps at four sites in New York state in 2003–2004. Trap placement and efficacy of chickens, Gallus gallus domesticus L., as bait compared with house sparrows, Passer domesticus L., an important reservoir of West Nile virus (family Flaviviridae, genus Flavivirus, WNV), was evaluated. Each site had a chicken-baited trap near ground level (≈1.5 m) and in the tree canopy (≈9 m), and a house sparrow-baited trap at ground level and canopy level. Each trap allowed mosquito access to birds on one end, and an inner mesh screen blocked bird access on the other end. The two most abundant mosquitoes, Culex restuans Theobald and Culex pipiens pipiens L., were differentiated using molecular characters. In 2003, Cx. restuans and Cx. p. pipiens made up 88% of total mosquito catch. In 2004, Cx. restuans comprised 43% of total catch and Cx. p. pipiens comprised 33%. The remaining species representing at least 1% of total catch were Ochlerotatus trivittatus (Coquillett), Coquilletidia perturbans (Walker), and Culiseta morsitans (Theobald). Capture rates were similar for chicken and house sparrow-baited traps; however, significantly more mosquitoes were captured in the canopy for both bird species. Cx. restuans preferred canopy traps, whereas equal numbers of Cx. p. pipiens were captured at ground and canopy levels. Mosquitoes were more likely to escape (74%) when excluded from birds than when allowed free access to birds (54%). Sentinel bird surveillance for WNV can be improved by trapping in the tree canopy in addition to ground level to capture the most important avian vectors.

More than 20 species of fleas in California are implicated as potential vectors of Yersinia pestis. Extremely limited spatial data exist for plague vectors—a key component to understanding where the greatest risks for human, domestic animal, and wildlife health exist. This study increases the spatial data available for 13 potential plague vectors by using the ecological niche modeling system Genetic Algorithm for Rule-Set Production (GARP) to predict their respective distributions. Because the available sample sizes in our data set varied greatly from one species to another, we also performed an analysis of the robustness of GARP by using the data available for flea Oropsylla montana (Baker) to quantify the effects that sample size and the chosen explanatory variables have on the final species distribution map. GARP effectively modeled the distributions of 13 vector species. Furthermore, our analyses show that all of these modeled ranges are robust, with a sample size of six fleas or greater not significantly impacting the percentage of the in-state area where the flea was predicted to be found, or the testing accuracy of the model. The results of this study will help guide the sampling efforts of future studies focusing on plague vectors.

Field and laboratory studies were performed to verify whether Culex theileri Theobald functions as a natural vector of Dirofilaria immitis (Leidy) on Madeira Island, Portugal. CO₂-baited light traps (EVS traps) were use to sample mosquitoes monthly basis between February 2002 and February 2003 in the area of Quebradas (Funchal). Three mosquito species were captured, including 58 Culex pipiens L., 790 Cx. theileri, and three Culiseta longiareolata (Macquart). Only C. theileri tested positive for D. immitis. The presence of this filarial worm was detected by direct observation, infectivity assay dissection technique, and polymerase chain reaction methods. Infected mosquitoes were recovered in October and December 2002 and January 2003. These data provide evidence that Cx. theileri could be the main vector of D. immitis in Funchal, Madeira.

Anopheles aquasalis Curry, a coastal malaria vector with a Neotropical distribution, was collected from Belém, Pará state, Brazil, and 500 adults per cage were maintained at the Instituto Evandro Chagas insectary at 26–30°C and 80–90% RH, where they fed on a 10% domestic sugar solution and blood from white mice. Oviposition of the parental generation (P) occurred in fresh water in dark cups introduced into mosquito cages. After eclosion, 100 larvae per pan were reared in artesian well water (salinity 0.04 g liter⁻¹) and fed ground fish food until pupation. After force mating in the F₁ generation, the eight subsequent generations were free mating. Mean larval mortality was <1%, and the mean developmental time from eclosion to emergence was 7.7 d (F₁), 7.6 d (F₂), 8 d (F₃), and 7.5 d (F₄). The maximum daily production of pupae (from the fourth generation on) occurred on day 6 postoviposition.

Trench fever is a body louse-borne disease caused by Bartonella quintana Brenner. The recent status of louse infestation in Nepalese children is not well known. We collected head and body lice, Pediculus humanus capitis De Geer and Pediculus humanus humanus L., respectively, from 30 children, including 11 cases of double infestation with both head and body lice. Detection of B. quintana in both louse species identified was carried out by polymerase chain reaction (PCR). PCR products with B. quintana DNA sequences were detected in both head and body lice from two children as well as in body lice derived from two other children. These results demonstrate that head lice may also play a role in the transmission of trench fever.

In laboratory tests, ethyl acetate extracts of Hyptis suaveolens Poit. from Guinea-Bissau and Rhododendon tomentosum (Stokes) H. Harmaja (formerly Ledum palustre L.) and Myrica gale L. significantly reduced probing activity of Aedes aegypti (L.). In the field in southern Sweden, extracts of leaves of R. tomentosum, M. gale, and Achillea millefolium L. significantly reduced biting by Aedes mosquitoes. Volatile compounds from M. gale, R. tomentosum, A. millefolium, and H. suaveolens were collected by solid phase microextraction (SPME). Alternatively, compounds in the plants were subjected to extraction by organic solvents of different polarities or by steam distillation and collection by SPME. Compounds collected were identified by gas chromatography-mass spectrometry. Leaves of H. suaveolens contained mainly β-caryophyllene, bergamotene, and terpinolene. The volatile fraction of an ethyl acetate extract of H. suaveolens was collected by SPME and included β-caryophyllene, (−)-sabinene, β-pinene, limonene, α-pinene, and bergamotene. The main volatiles detected were α-pinene, α-phellandrene, myrcene, and limonene from M. gale leaves or inflorescences; p-cymene, sabinene, and terpinyl acetate from leaves of R. tomentosum; and (−)-germacrene D, β-pinene, sabinene, and α-pinene from A. millefolium leaves or inflorescences. The selected plant species contained numerous volatiles known to have insecticidal, acaricidal, “pesticidal,” and/or insect repellent properties.

We investigated whether host-seeking nymphs and adults of the western blacklegged tick, Ixodes pacificus Cooley & Kohls, the primary vector of Lyme disease spirochetes in far-western North America, are infected naturally with relapsing-fever group spirochetes in Mendocino County, California. Relapsing-fever group borreliae were detected in four (1.7%) of 234 nymphal and two (0.7%) of 282 adult host-seeking I. pacificus ticks by polymerase chain reaction and sequence analysis of the 16S rRNA and flagellin genes, respectively, exhibiting 99 and 98.5% sequence homology to Borrelia miyamotoi Fukunaga. Phylogenetic analysis based on these two genes revealed that the borreliae detected in these ticks belong to the relapsing-fever group and that these are closely related to, if not identical with, B. miyamotoi.