Mosquito seasonality and arboviral disease incidence in Murray Valley, southeast Australia

Abstract. Adult female mosquito populations were monitored at weekly intervals during spring-autumn (November-March) for 4 years (1991-95) using dry-ice-baited light traps at forty sites in the Murray Valley of Victoria, Australia. Among twenty species of mosquitoes collected, Culex annulirostriswas the most abundant (66.6 ± 9.3%) followed by Cx australicus(15.3 ± 7.7%). From a total of 476, 682 mosquitoes collected, nearly all were females and only 1295 (0.27%) were males. Mosquito population densities were generally higher in 1992-93 and 1993-94 seasons than in 1991-92 and 1994-95 seasons. Greatest densities of Cx annulirostrisand Cx australicusoccurred in 1992-93, coinciding with outbreaks of Ross River (RR) and Barman Forest (BF) arboviruses causing human polyarthritis. In the majority of shires, Cx australicuswas the predominant species from spring to early summer (November and December), then was replaced by Cx annulirostrisfrom mid-summer to autumn (January-April). In three shires, Aedes bancroftianusand Ae.sagaxpredominated during the early part of the season. Densities of both Cx annulirostrisand Cx australicuswere related to temperature. Cx australicusadults were found to be trapped when the mean ambient temperature exceeded 6d?C, with peak population recorded at 20d?C. Cx annulirostrisadult density increased when the mean temperature rose above 12d?C, reaching a peak during February and March when temperature exceeded 25d?C. Cx annulirostrisdeclined rapidly from April onwards, with no adult activity evident from May to November. Population densities of Aedesspp. were generally less than reported from earlier studies, possibly due to lower rainfall in spring and summer as well as reduced flood irrigation practices. In each year, a significant correlation was detected between Cx annulirostrisdensity and RR virus incidence in humans. As Cx annulirostrisis the predominant local mosquito species and feeds on a wide spectrum of hosts including man, it seems likely that Cx annulirostrisis the major vector of RR in the inland Murray Valley region.     

Laboratory and field evaluation of commercial repellent formulations against mosquitoes (Diptera: Culicidae) in Queensland, Australia

Abstract  Laboratory tests of commercial repellent formulations were conducted against Anopheles farauti Laveran, Culex annulirostris Skuse, Ochlerotatus vigilax (Skuse) and Stegomyia aegypti (L.). The majority of repellent formulations tested contain N,N ,-diethyl- 3 -methylbenzamide (also known as diethyl- m -toluamide, commonly called deet). Two formulations containing picaridin (1-piperidinecarboxylate acid, 2-(2-hydroxyethyl)-1-methylpropylester, also known as KBR 3023), one containing ethyl butylacetylaminopropionate (EBAP) and two formulations containing essential oils (e.g. Citronella oil) were also tested. In the laboratory tests, repellent formulations containing deet provided the best protection, and picaridin and EBAP also provided good protection. Citronella oil provided only limited protection. Two field trials to compare commercially available repellent formulations containing picaridin and deet against mosquitoes at Redcliffe, Queensland, were conducted. In the first, Autan Repel, containing 9.3% picaridin, RID, containing 10% deet, and Bushman Ultra, containing 80% deet in a gel, were compared. In the second, Autan Repel Army 20, containing 19.2% picaridin, OFF! Skintastic, containing 7% deet, and Aerogard, containing 12% deet, were compared. The predominant mosquito in both tests was Cx. annulirostris . Bushman provided >95% protection against all mosquitoes for at least 8 h when tests ceased. The other deet repellents also provided good protection against mosquitoes, with RID providing 5 h, Skintastic 4 h and Aerogard 2 h protection. The Autan repel (9.3% picaridin) provided >95% protection for 3 h, and Autan Repel Army (19.2% picaridin) provided 4 h protection. These studies have shown that commercial formulations of both deet and picaridin provide good protection against Cx. annulirostris , an important vector of arboviruses in Australia.     

The vector competence of Culex annulirostris, Aedes sagax and Aedes alboannulatus for Murray Valley encephalitis virus at different temperatures

Culex annulirostris Skuse, colonized from Brisbane, Queensland, and Mildura, Victoria, Australia, were effective vectors of Murray Valley encephalitis virus at 20, 27 and 32-35 degrees C with full extrinsic incubation periods of 15, 10 and 4 days respectively. At 20 degrees C, 7-11 days post-infection, transmission by the Mildura colony (0-20%) was less efficient than the Brisbane colony (30-70%) but both were capable of 75-100% transmission after longer extrinsic incubation periods. Discriminant analysis of body and salivary gland titres showed that these were not satisfactory indicators of transmission. Wild-caught Aedes sagax (Skuse) and Cx annulirostris from the Murray Valley showed equal competence, but Aedes alboannulatus (Macquart) was a poor vector. The results provide data on rural amplification of Murray Valley encephalitis virus during spring and suggest that further work on the potential of Ae. sagax as a natural vector is warranted.     

New Records and Reference Collection of Mosquitoes (Diptera: Culicidae) on Jeju Island, Republic of Korea

Mosquito collections were carried out during 2003‐2004 on Jeju Island, Republic of Korea. Eighteen species of mosquitoes in 7 genera were collected, including three new records, Culex (Culex) mimeticus, Culex (Culiciomyia) sasai and Ochlerotatus (Finlaya) nipponicus. Based on the available information, a total of 28 species representing 7 genera have been recorded from Jeju Island. Larval habitat characteristics, collection sites, bionomics, and vector potential for each of these species are described.     

Mosquito (Diptera: Culicidae) fauna in inland areas of south-west Western Australia

Abstract  Previously existing sources of data regarding mosquitoes in inland areas of south-west Western Australia are few and qualitative in nature. This is the first attempt to quantitatively investigate mosquito fauna in this region. The existing data are reviewed and compared with the results of the quantitative surveys presented in this paper. Temporal comparisons appear to indicate that mosquito community structure in the region may have changed since the initial surveys in the 1950s from a combination of freshwater-breeding species towards a strong dominance of Aedes camptorhynchus (Thomson), a major vector of Ross River virus in southern Australia. It is speculated that this shift may have been brought about by the increasing area and severity of dryland salinity in the region over the last century, and also may increase the potential for Ross River virus disease transmission.     

Development time and survival of Verrallina funerea (Theobald) (Diptera: Culicidae) immatures and other brackish water mosquito species in southeast Queensland, Australia

Abstract  Verrallina funerea (Theobald) is a brackish water mosquito that is recognised as an important pest and vector in southeast Queensland, Australia. Immature development time and survival of Ve. funerea was defined in the laboratory in response to a range of temperatures (17–34°C) and salinities (0–35 parts per thousand (p.p.t)). The expression of autogeny in this species was also assessed. Salinity only had a slight effect on mean development time from hatching to adult emergence (7.0–7.4 d at salinities of 0, 17.5 and 31.5 p.p.t) and survival was uniformly high (97.5–99.0%). Mean development times were shorter at 26, 29 and 32°C (7.0, 6.8 and 6.8 d, respectively) and longest at 17°C (12.2 d). The threshold temperature ( t ) was 5.8°C and the thermal constant ( K ) was 142.9 degree-days above t . Survival to adulthood decreased from >95% (at 17–29°C) to 78% (at 32°C) and 0% (at 34°C). No expression of autogeny was observed. Immature development times of Ve. funerea , Ochlerotatus vigilax (Skuse) and Oc. procax (Skuse) were then determined under field conditions at Maroochy Shire. Following tide and rain inundation, cohorts of newly hatched larvae were monitored daily by dipping, and time until pupation was noted. Tidal inundation triggered hatching of Ve. funerea and Oc. vigilax larvae whereas Oc. procax larvae were found only after rain inundation. Estimates of Ve. funerea and Oc. vigilax field development times were similar (8–9 d) while Oc. procax development time was slightly longer (9–10 d). Based on these survey results, control activities targeting Ve. funerea must be initiated 4 d (if using Bacillus thuringiensis var. israelensis de Barjac) or 5 d (if using s -methoprene) after inundation. However, Casuarina glauca Sieber canopy and branchlets covering breeding habitats may present a problem for the penetration of such treatments.     

Molecular identification of mosquitoes (Diptera: Culicidae) in southeastern Australia

DNA barcoding is a modern species identification technique that can be used to distinguish morphologically similar species, and is particularly useful when using small amounts of starting material from partial specimens or from immature stages. In order to use DNA barcoding in a surveillance program, a database containing mosquito barcode sequences is required. This study obtained Cytochrome Oxidase I (COI) sequences for 113 morphologically identified specimens, representing 29 species, six tribes and 12 genera; 17 of these species have not been previously barcoded. Three of the 29 species ─ Culex palpalis, Macleaya macmillani, and an unknown species originally identified as Tripteroides atripes ─ were initially misidentified as they are difficult to separate morphologically, highlighting the utility of DNA barcoding. While most species grouped separately (reciprocally monophyletic), the Cx. pipiens subgroup could not be genetically separated using COI. The average conspecific and congeneric p‐distance was 0.8% and 7.6%, respectively. In our study, we also demonstrate the utility of DNA barcoding in distinguishing exotics from endemic mosquitoes by identifying a single intercepted Stegomyia aegypti egg at an international airport. The use of DNA barcoding dramatically reduced the identification time required compared with rearing specimens through to adults, thereby demonstrating the value of this technique in biosecurity surveillance. The DNA barcodes produced by this study have been uploaded to the ‘Mosquitoes of Australia–Victoria’ project on the Barcode of Life Database (BOLD), which will serve as a resource for the Victorian Arbovirus Disease Control Program and other national and international mosquito surveillance programs.     

Timing of host-seeking behaviour of the mosquitoes Anopheles annulipes sensu lato Walker and Coquillettidia linealis (Skuse) (Diptera: Culicidae) in the Murray River Valley, South Australia

Abstract  The timing of host-seeking behaviour was studied in two mosquito species, Anopheles annulipes sensu lato and Coquillettidia linealis at Mundic Creek, near the Upper River Murray in South Australia. Hourly sampling using carbon dioxide-baited traps revealed consistent patterns in the timing of host-seeking. Coquillettidia linealis displayed a very large peak in host-seeking activity at sunset, followed by a much reduced level throughout the night and a smaller peak at sunrise. Anopheles annulipes s.l. differed by displaying a significantly smaller peak at sunset, followed by higher amount of activity throughout the night compared with Cq. linealis . The two taxa, while both crepuscular and nocturnal, spread their host-seeking effort out differently throughout the night, indicating that they have different resource requirements.     

Characterization of microsatellite markers in the mosquito Ochlerotatus caspius (Diptera: Culicidae)

Seven polymorphic microsatellite loci were developed for the mosquito species Ochlerotatus caspius, using an enriched genomic library. The number of alleles per locus varied between two and 11; the expected heterozygosity (H_E) ranged from 0.18 to 0.77. These microsatellite primers should prove useful for population genetic studies of this mosquito species.     

Aquatic thermal conditions predict the presence of native and invasive rock pool Aedes (Diptera: Culicidae) in the southern Appalachians,U.S.A.

The native rock pool mosquito, Aedes atropalpus (Coquillett), and the invasive Aedes japonicus (Theobald) have been found in many types of artificial and natural containers throughout North America. Little is known about the ecology of these two species in habitats where they co‐occur, although multiple investigators have reported the decline of the native species concurrent with the introduction and spread of the invasive species. Here we report the results of riverine rock pool collections (n=503) in the southern Appalachian Mountains between 2009‐2015. Surface water temperatures strongly predicted the presence of each species across a broad range of observed temperatures (11‐39.8° C). For every unit of increase in temperature (°C) the odds of collecting Ae. atropalpus larvae increased by 0.34 while the odds of collecting Ae. japonicus larvae decreased by 0.28. No Ae. japonicus larvae or pupae were collected at temperatures greater than 36° C; however, immature Ae. atropalpus were found in rock pools with temperatures up to 39.8° C. In contrast, Ae. japonicus were highly abundant in cooler rock pools (<17° C) where Ae. atropalpus were infrequent or absent. Our findings suggest that in spite of the successful invasion by Ae. japonicus, Ae. atropalpus remains well established in the southern Appalachian Mountains. Given the strong correlation of temperature with the presence of the two species and the contrasting absence of each species at observed temperature extremes, the role of thermal conditions should be carefully explored in the context of other ecological factors likely influencing the range and abundance of these mosquitoes.     

Tires as larval habitats for mosquitoes (Diptera: Culicidae) in southern Manitoba, Canada

In 2003, a survey at waste management grounds and tire dealerships was conducted to determine the species composition of mosquitoes in tires in southern Manitoba, Canada. Over 25% of the 1,142 tires sampled contained a total of 32,474 mosquito larvae and pupae. Culex restuans made up at least 95% of the larvae collected for each month of the summer. Culiseta inornata and Culex tarsalis reached their greatest numbers in July and August, respectively, though they were never abundant. Ochlerotatus triseriatus was also found but never reached more than 1% of the total larvae collected in any given month. Mosquito prevalence was more than three times greater in August (36.1%) than in June (11.7%). Orientation affected prevalence of mosquitoes in tires: 31.4% of vertical tires (tires standing on their treads) contained mosquitoes, whereas mosquitoes were found in only 18.9% of horizontal tires (tires parallel to the ground). Tires in the eastern region of Manitoba contained mosquitoes more often (61.7%), irrespective of date, than Winnipeg (25.9%), the central region (29.1%), or the western region (19.8%). Mosquito prevalence was similar across three size categories of tires, car tires (18.8%), truck tires (19.8%), and semi-trailer tires (26.7%), though tractor tires (47.8%) contained significantly more mosquitoes than tires in the other categories.     

Existence of Two Acetylcholinesterases in the Mosquito Culex pipiens (Diptera: Culicidae)

Abstract: Two acetylcholinesterases (AChEs), AChE1 and AChE2, differing in substrate specificity and in some aspects of inhibitor sensitivity, have been characterized in the mosquito Culex pipiens . The results of ultracentrifugation in sucrose gradients and nondenaturing gel electrophoresis of AChE activity peak fractions show that each AChE is present as two molecular forms: one amphiphilic dimer possessing a glycolipid anchor and one hydrophilic dimer that does not interact with nondenaturing detergents. Treatment by phosphatidylinositol-specific phospholipase C converts each type of amphiphilic dimer into the corresponding hydrophilic dimer. Molecular forms of AChE1 have a lower electrophoretic mobility than those of AChE2. However, amphiphilic dimers and hydrophilic dimers have similar sedimentation coefficients (5.5S and 6.5S, respectively). AChE1 and AChE2 dimers, amphiphilic or hydrophilic, resist dithiothreitol reduction under conditions that allow reduction of Drosophila AChE dimers. In the insecticide-susceptible strain S-LAB, AChE1 is inhibited by 5 × 10⁻⁴ M propoxur (a carbamate insecticide), whereas AChE2 is resistant. All animals are killed by this concentration of propoxur, indicating that only AChE1 fulfills the physiological function of neurotransmitter hydrolysis at synapses. In the insecticide-resistant strain, MSE, there is no mortality after exposure to 5 × 10⁻⁴ M propoxur: AChE2 sensitivity to propoxur is unchanged, whereas AChE1 is now resistant to 5 × 10⁻⁴ M propoxur. The possibility that AChE1 and AChE2 are products of tissue-specific posttranslational modifications of a single gene is discussed, but we suggest, based on recent results obtained at the molecular level in mosquitoes, that they are encoded by two different genes.     

The 2000 epidemic of Rift Valley fever in Saudi Arabia: mosquito vector studies

In mid-September 2000, Rift Valley fever (RVF) virus was diagnosed as the cause of infection in humans and livestock in Jizan Region, Saudi Arabia. This is the first time that this arbovirus has been found outside Africa and Madagascar. Collections of mosquitoes (Diptera: Culicidae) were therefore undertaken (from 25 September to 10 October) at eight sites during the epidemic to obtain mosquitoes for attempted RVF virus isolation. Among 23 699 mosquito females tested, six isolations of RVF virus were made from 15 428 Culex (Culex) tritaeniorhynchus Giles and seven from 8091 Aedes (Aedimorphus) vexans arabiensis Patton [corrected]. Minimum mosquito infection rates per 1000 at sites with infected mosquitoes were 0.3-13.8 Cx. tritaeniorhynchus and 1.94-9.03 Ae. v. arabiensis. Viral activity moved northwards as collecting was in progress and collectors 'caught up' with the virus at the two most northerly sites on the last two trapping evenings. Other species occurred in small numbers and were identified but not tested. Both Cx. tritaeniorhynchus and Ae. v. arabiensis were susceptible to RVF virus and transmitted between hamsters, and an additional quantitative test with Cx. tritaeniorhynchus showed that 71-73% of mosquitoes became infected after ingesting 6.9-7.9 log10 FFU/mL of virus; transmission rates were 10% (post-infection day 14) and 26% (post-infection day 20). It was concluded that both species were vectors on grounds of abundance, distribution, preference for humans and sheep, the virus isolations and vector competence tests.     

Molecular composition of the chromosome 2 pericentric heterochromatin in malarial mosquitoes (Diptera,Culicidae)

Rich in repeated DNA sequences and poor in genes, the heterochromatin is an important functional part of the eukaryotic genome. Heterochromatin exhibits high evolutionary variability, which was revealed on the cytological and molecular levels in malarial mosquito species from the Anopheles maculipennis complex. In this connection, investigation of the heterochromatin molecular composition in species of this complex is of interest.     

Updated checklist of the mosquitoes (Diptera: Culicidae) of Croatia

Improvement of morphological and molecular identification methods allows the detection of new species of mosquitoes. The mosquito fauna of Croatia currently includes 52 species, belonging to eight genera, including Anopheles (12 species), Aedes (24 species), Coquillettidia (one species), Culex (seven species), Culiseta (six species), Orthopodomyia (one species), and Uranotaenia (one species). This is an updated checklist, which includes five new species found in Croatian mosquito fauna. Two of these are invasive mosquito species, Aedes albopictus (Skuse, 1895) and Aedes japonicus (Theobald 1901), which are spreading across Europe and Croatia. The other three species, Culex laticinctus (Edwards 1913), Culex torrentium (Martini 1925), and Anopheles daciae (Linton, Nicolescu & Harbach 2004) are autochthonous species which haven't been recorded so far. Since there are several more invasive species spreading across Europe, we assume that this is not the final list.