Laboratory evaluation of Toxorhynchites splendens (Diptera: Culicidae) for predation of Aedes albopictus mosquito larvae

Biology of the mosquito Toxorhynchites splendens (Wiedemann) was studied in the laboratory to provide baseline data for using the predatory larvae of this species against those of Aedes albopictus (Skuse) in a biological control programme. The mean incubation time of Tx.splendens eggs was 43.8 h and the time required for newly-hatched larvae to initiate predation was 2.5 h. Mean numbers of prey larvae consumed and killed by each Tx.splendens larva totalled 389 and 345 respectively. The larval period of Tx.splendens was not significantly different for rearing individually or in groups of nine, with equal prey density, and duration of larval development was proportional to prey density. In mass rearing, larval cannibalism was usually observed during days 1-3 post-eclosion. The incidence of cannibalism decreased sharply on the fourth day after hatching when some larvae became fourth-instar. Adult female Tx.splendens usually commenced oviposition on day 4 after emergence. The number of eggs laid daily increased on day 7 and the peak oviposition of 6.3 eggs/female/day occurred on day 11. When oviposition containers were provided only intermittently, gravid females of Tx.splendens scattered most of their eggs on the dry floor of the cage. Viability of eggs laid by females aged 4-14 days was high (60-90%) but decreased to less than 40% as the females aged.     

Changes in the fat body during the post-embryonic development of the predator Toxorhynchites theobaldi (Dyar & Knab) (Diptera: Culicidae)

Several studies have focused on understanding the biochemistry and morphology of the fat body of the hematophagous mosquito Aedes aegypti (L.) (Diptera: Culicidae). In contrast, few studies, if any, have focused on morphological characters of the fat body in other mosquitoes, especially non-hematophagous taxa such as the culicid Toxorhynchites. Larvae of Toxorhynchites prey upon the larvae of other mosquito species and are used in vector mosquito control. We investigated aspects of the fat body trophocytes, including the morphometric analyses of the lipid droplets, protein granules and nuclei, during Toxorhynchites theobaldi (Dyar & Knab) post-embryonic development. Following the body weight increase from larval stage L2 to L4, the size of lipid droplets within the trophocytes also increase, and are likely the result of lipogenesis. Lipid droplets decrease in size during L4 to the female pupal stage and increase once again during the period from newly-emerged to mature adult females. Protein granules are observed for the first time in female pupae, and their appearance might be related to protein storage during metamorphosis. The size of the nucleus of trophocytes also increases during larval development, followed by a decrease during metamorphosis and an additional increase as adult female ages. In conclusion, the morphology of the fat body of T. theobaldi changes according to the developmental stage. Our study provides for the first time important insights into T. theobaldi fat body development and contributes to understand this species biology.     

The mosquitoes (Diptera: Culicidae) of Tabasco,Mexico

To document and update the mosquito species of Tabasco, Mexico, field collection trips were conducted in the two physiographic regions of Tabasco: the coastal plain of the southern gulf and the mountains of Chiapas and Guatemala. Mosquitoes were collected as immature and adult stages during the dry and rainy seasons from 2014 through 2015. Additionally, the Reference Collection of Arthropods of Medical Importance (CAIM‐InDRE) containing mosquitoes of Tabasco was re‐examined. In total, 4,913 specimens were collected and examined, which are divided into seven tribes, 18 genera, 27 subgenera, and 104 species. Of these, one genus (Shannoniana Lane and Cerqueira), two subgenera (Georgecraigius Reinert, Harbach and Kitching, and Carrollia Lutz), and 21 species are new records for the mosquito fauna of Tabasco. Culex metempsytus Dyar is a new record for Mexico and Wyeomyia jocosa (Dyar and Knab) is removed from the Mexican mosquito fauna. Seventeen species historically reported were not found in the field collections conducted here. Taxonomic notes, new distribution limits, and comments about the medical importance of species of mosquitoes of Tabasco are discussed. Tabasco is the second state in Mexico with the largest mosquito richness (104 species), followed by Veracruz with 139 species.     

Effect of cibarial armature of mosquitoes (Diptera,Culicidae) on blood-meal haemolysis

Cibarial armatures of mosquitoes act mechanically on the ingested erythrocytes producing the haemolysis of part of the blood meal. The extent of haemolysis, in blood meals examined immediately after ingestion, is less than 5% in mosquitoes without cibarial armature or with “primitive” types of cibarial armature, while up to 50% haemolysis is observed in mosquitoes with more highly evolved types of cibarial armature. Such high levels of haemolysis are produced by the passage of the blood through the mosquito head but not by the incubation of the blood with salivary gland and midgut homogenates from the same mosquito or by injection of the blood into the intestine via the anus.     

Prey finding behavior and mechanosensilla of larval Toxorhynchites brevipalpis Theobald (Diptera : Culicidae)

Behavioral experiments demonstrated that starved 3rd-instar Toxorhynchites brevipalpis (Diptera : Culicidae) will attack a glass probe in response to vibrations alone. Frequencies in the range of 80–200 Hz elicited 85% of the attacks. The observation that most attacks occurred while the larva was drifting forward towards the probe, substantiates that the predatory behavior of these organisms is basically opportunistic.The main setae on the thorax and abdomen arise from setal support plates and are of 4 general morphological types: dendritic, smooth, spinulate spiniform, and unbranched aciculate setae. The numbers and lengths of the setae on 3rd- and 4th-instar larvae are given. Each seta is innervated by a bipolar neuron with a tubular body in the dendrite, a characteristic of mechanosensilla. By virtue of their abundance and location, it seems probable that one or more types of these setae sense the water vibrations capable of eliciting an attack response.The tubular bodies of the 4 types of setae are somewhat unusual in that they lack electron-dense material, except near sites of attachment of the microtubules to the dendritic membrane, yet possess a large number of complexly arranged microtubules. Among the setal types, variations were observed in the prominence of microtubular attachment sites, proportion of tubules associated with the sites, and orientation of microtubules. In spinulate spiniform setae, the arrangement of microtubules varies within the same tubular body; peripherally most of the microtubules are in one direction with little convergence and have only one distinct attachment site, whereas medially they converge considerably resulting in 2 groups, each associated with its own well-defined site.The attachment sites, which presumably through linkage with the dendritic membrane provide a means of initiating depolarization, are associated with the distal ends of almost all of the microtubules. This suggests that in these tubular bodies depolarization may be initiated through mechanical force acting along the length of the microtubules, that is, stretching and/or compression.     

Laboratory experiments on factors affecting oviposition site selection in Toxorhynchites amboinensis (Diptera: Culicidae), with a report on the occurrence of egg cannibalism

Laboratory experiments tested the effects of water quality and the presence of conspecific and heterospecific immatures on oviposition by Toxorhynchites amboinensis (Doleschall). The females showed a highly significant preference for oak leaf infusion water over distilled water. When twenty starved third and fourth instar Tx.amboinensis larvae were present in the water, substantially fewer eggs were counted from pots containing these conspecifics, than from controls in which no larvae were present. Numbers of eggs from pots containing starved second instar larvae did not differ significantly from controls. Observations of larval behaviour while oviposition was occurring suggested that egg numbers were reduced in containers because of egg cannibalism with third and fourth instar larvae, and not because the larvae caused a deterrent effect. Subsequent experiments confirming the occurrence of substantial egg cannibalism by third and especially fourth instar larvae are described. As with larvae, the presence of Tx.amboinensis pupae in the water had little effect on oviposition. If placed in the water 24 h prior to test, pupae very slightly enhanced its attractiveness, but if introduced immediately before test there appeared to be a slight deterrent effect. With heterospecific larvae, twenty fourth instar Ae.aegypti larvae introduced into infusion water 24 h before test rendered the water slightly attractive, while water in which Ae.aegypti larvae had been reared for 48 h proved highly repellent.     

Phylogeny of mosquitoes of tribe Culicini (Diptera: Culicidae) based on morphological diversity

Harbach, R. E., Kitching, I. J., Culverwell, C. L., Dubois, J. & Linton, Y.‐M. (2012). Phylogeny of mosquitoes of tribe Culicini (Diptera: Culicidae) based on morphological diversity. —Zoologica Scripta, 41, 499–514. Relationships among taxa of the mosquito tribe Culicini are explored using 169 morphological characters from 86 exemplar species representing the four genera and 26 subgenera of Culicini, most species groups and subgroups of subgenus Culex and an outgroup of five species from five other tribes. We analysed the data set, with multistate characters treated as unordered, under implied weights with values of K ranging from 1 to 20, implemented by TNT. Each analysis, except K = 4, produced a single most parsimonious (fittest) cladogram (MPC). The topology of the ingroup was identical for K = 6–11, whereas the MPCs for K = 14–20 differed only in the position of a single species, which occupied the same position in the K = 16 and K = 6–11 topologies. The K = 9 and K = 16 trees were given further consideration. In both these cladograms, Lutzia is sister to a clade comprising genera Culex, Deinocerites and Galindomyia. The two topologies have 13 clades in common, but their arrangements differ primarily because of Culex (Culex) duttoni acting as a ‘rogue’ taxon. We evaluated the effect of removing this species from the K = 9 and K = 16 analyses and chose the refined K = 9 topology as the best hypothesis of relationships within Culicini. Genus Culex is not monophyletic because it includes Deinocerites and Galindomyia as derived members of the New World subgenera. With the exception of subgenera Culex, Eumelanomyia and Neoculex, there is strong support for the monophyly of all genera and subgenera. Subgenus Culex would be monophyletic were four other subgenera included and three other taxa (the Afrotropical Cx. duttoni, Neotropical Cx. apicinus and the Australasian Atriceps Group) excluded.     

Inversion polymorphism and ecological specialization of malaria mosquitoes (Diptera, Culicidae) in Karelia

The species composition and inversion polymorphism were studied in malaria mosquito larva hemipopulations of Karelia. Three malaria mosquito species??Anopheles messeae, A. beklemishevi, and A. maculipennis??were found in the region. The northern boundary of their range is at 65°N. The greatest species diversity was observed in biotopes of the central region. Within-species chromosome polymorphism was observed in larva populations of all three species. For A. messeae, maximum karyotype diversity indices were established for the southern and northern regions of Karelia.     

Biological control of container-breeding mosquitoes, Aedes albopictus and Culex quinquefasciatus, in a Japanese island by release of Toxorhynchites splendens adults

To control container-breeding mosquitoes in the small island of Minnajima (0.56 km2), northern Okinawa, Japan, laboratory-reared adults (aged 7-10 days) of Toxorhynchites splendens (Palawan strain), a mosquito with predatory larvae, were released repeatedly during 1984, 1986 and 1987. Thirteen species of mosquitoes (Diptera: Culicidae) occurred in artificial containers, ground pools or crab-holes on the island, the predominant species being Aedes (Stegomyia) albopictus and Culex (Culex) quinquefasciatus. Predatory mosquito larvae of Culex (Lutzia) fuscanus and Cx (Lt.) halifaxii were found commonly in wet containers. In the first year of study, during a period of 54 days from 13 May to 5 July 1984, totals of 879 female and 806 male adults of Tx.splendens were released on six occasions. Similarly, between 29 April and 30 August 1986, totals of 2920 female and 2878 male adult Tx.splendens were released. In the third study year, totals of 2041 female and 1783 male Tx.splendens were released on eight occasions during 199 days from 23 April to 7 November 1987. After adult releases at two sites, the immature stages of Tx.splendens were found in 164 out of 502 traps in 1984, 421 out of 933 traps in 1986, and 151 out of 502 traps in 1987. The number of immatures of Tx.splendens present in each trap varied from 1 to 40 in 1984, 1 to 29 in 1986 and 1 to 9 in 1987. Numbers of immatures of the target species found in the traps during August-September averaged 71.9/trap/month in 1984, 114.7/trap/month in 1986 and 36.0/trap/month in 1987, significantly less in the traps with Tx.splendens than in those without them. The present field studies indicated that, in this small island, approximately 250 adult female and 200 male Tx.splendens per month should be released from April to November, and the releases should be carried out every year, in order to control effectively the target mosquitoes Ae.albopictus and Cx quinquefasciatus breeding in artificial containers in Minnajima.     

Effect of temperature on the development and consumption of Phaenochilus kashaya (Coleoptera: Coccinellidae), a predator of the cycad aulacaspis scale,Aulacaspis yasumatsui

The cycad aulacaspis scale (CAS), Aulacaspis yasumatsui Takagi (Hemiptera: Diaspididae), is a serious pest of ornamental cycads in the southeastern United States. In Florida, CAS was first reported in Miami in 1996 and is now present in 43 Florida counties. Chemical control of CAS can be effective, but it is expensive, and insecticides must be frequently and regularly applied. Recent surveys of natural enemies in Thailand identified a new potential biological control agent of CAS. The objective of this study was to examine the temperature requirements and consumption rates of Phaenochilus kashaya Giorgi and Vandenberg (Coleoptera: Coccinellidae). Survival, developmental time and number of scales consumed by P. kashaya were measured at four constant temperatures (20, 25, 30 and 35°C). In addition, adult longevity and consumption were recorded at three constant temperatures (20, 25 and 30°C). Phaenochilus kashaya completed development to adulthood at temperatures ranging from 20 to 30°C. The highest survival rate (48%) was obtained at 25°C, and the accumulated degree-days for this species was 714. Phaenochilus kashaya is a voracious predator of CAS, consuming 380 scales during the larval stage and an estimated 4700 scales during the adult stage. Adult longevity varied from 59 days at 30°C to 220 days at 20°C for females, and 31 days at 30°C to 148 days at 20°C for males. The intrinsic rate of increase (r _m) was 0.34 at 20°C, which is high compared with other coccinellid species. According to this study, P. kashaya has the potential to be an effective biological control agent of the CAS in Florida.     

Sequence and secondary structure comparisons of ITS rDNA in mosquitoes (Diptera: Culicidae).

Sequences of the internal transcribed spacers (ITS1 and ITS2) of the mosquito Aedes aegypti, and the ITS2 of six related species, A. simpsoni, A. albopictus, A. vexans, A. triseriatus, Haemagogus mesodentatus, and Psorophora ferox are reported. Intraspecific variation in A. aegypti ITS1 is 1.07% among four clones from three individuals, and in the ITS2 is 1.17% among 15 clones from four individuals. In A. simpsoni, intraspecific ITS2 variation is 0.46% among 10 clones from a single individual. Alignment of the ITS2 sequence of the seven species reveals several homologous domains. Secondary structure predictions for the ITS2 region indicate that these domains base pair to form a core region central to several stem features. The sequence outside the ITS2 homologous domains tends to be GC-rich and characteristically slippage generated; these areas preserve or add to the stem length of the predicted secondary structures. These ITS2 intraspacer variable regions resemble previously described expansion segments of the 28S gene region. Evolutionary analysis of the ITS2 of these species, using both sequence and secondary structure information, leads to the prediction of divergence in the mosquito tribe Aedini that is not clearly reflected in current taxonomic designations.     

Predation potential of three flatworm species (Platyhelminthes: Turbellaria) on mosquitoes (Diptera: Culicidae)

We conducted a field survey for flatworms to select species as potential biological control agents against Aedes aegypti and Culex pipiens (Diptera, Culicidae) breeding in artificial containers. Laboratory experiments were performed to determine the daily predation rate, differential predation on each mosquito larval instar, selective predation on either A. aegypti or C. pipiens, and predator tolerance to water from artificial containers. Girardia anceps (Tricladida, Paludicola, Dugesiidae), Mesostoma ehrenbergii and Bothromesostoma cf. evelinae (Rhabdocoela, Typhloplanoida, Typhloplanidae) were found in temporary puddles and permanent pools. In the laboratory, they killed between 52% and 100% of immature mosquitoes coexisting in the same habitat. No preference of flatworms for mosquito preys was detected. Predation rate was related to predator size and instar of preys. Girardia anceps and B. evelinae survived after a dry period and when re-flooding occurred, they laid eggs. Tolerance to water from artificial containers was highest in G. anceps and this species could be a suitable predator to reduce mosquito populations from artificial containers using an inoculative approach.     

Phylogenetic analysis and temporal diversification of mosquitoes (Diptera: Culicidae) based on nuclear genes and morphology

Background  

Phylogenetic analyses provide a framework for examining the evolution of morphological and molecular diversity, interpreting patterns in biogeography, and achieving a stable classification. The generic and suprageneric relationships within mosquitoes (Diptera: Culicidae) are poorly resolved, making these subjects difficult to address.     

Eclosion rate, development and survivorship of Aedes albopictus (Skuse)(Diptera: Culicidae) under different water temperatures

In tropical areas, where vector insects populations are particularly numerous, temperature usually range between 25 degrees C and 35 degrees C. Considering the importance of such temperature variation in determining mosquitoes population dynamics, in this work the developmental, eclosion and survival rates of the immature stages of Aedes albopictus (Skuse) were compared under constant 25, 30 and 35 degrees C (using acclimatized chambers) and environmental (25 degrees C to 29 degrees C) temperatures. The hatching rate was considered as total number of larvae recovered after 24h. The development period as well as larval and pupal survival rate were evaluated daily. Eclosion rate was significantly higher under environmental temperature than under the studied constant temperatures, suggesting that temperature variation may be an eclosion-stimulating factor. The mean eclosion time increased with the temperature, ranging from 2.8h (25 degrees C) to 5.2h (35 degrees C). The larval period was greatly variable inside each group, although it did not differ significantly amongst groups (11.0 +/- 4.19 days), with individuals showing longer larval stages in water at 35 degrees C (12.0 +/- 4.95 days) and environmental temperature (13.6 +/- 5.98 days). Oppositely, survival was strongly affected by the higher temperature, where only one individual lived through to adult phase. The results suggest that population of Ae. albopictus from Recife may be adapting to increasing of environmental temperatures and that the limiting temperature to larval development is around 35 degrees C.     

Population dynamics of the immature stages of Aedes aegypti (Diptera: Culicidae), vector of dengue: a longitudinal study (1996-2000)

A four year study was conducted on a natural population of immature stages of Aedes aegypti after the re-invasion of Argentina by this vector in 1987. Thirty six plastic containers with 700 ml of dechlorinated water were placed in the La Plata Zoological Garden, La Plata, Argentina. A strip of filter paper around each container was added to facilitate egg counting. Eggs, larvae and pupae were counted weekly in each container from September, 1996 to August, 2000. After egg counting, papers were submerged to facilitate egg hatching and a new paper was placed in each container. Presence of A. aegypti immature stages was recorded from December-January to June during each of the four years of this study. In 1997, 13,105 eggs, 7,978 larvae and 1,476 pupae were registered with 54.7 % positive containers; during 1998, 8,194 eggs, 668 larvae and 142 pupae were recorded with 28.3 % positive containers; 13,510 eggs, 3,690 larvae and 743 pupae were registered during 1999 with 56.7 % positive containers; and 16,327 eggs, 4,669 larvae and 715 pupae during 2000 with 59.3 % of containers with presence of A. aegypti. Egg number and hatching rate were drastically reduced in 1998 when temperatures from December to May were 1 to 2.5 degrees C lower than the other years of this study. These colder than usual temperatures in the summer of 1998 were a consequence of the El Ni?o event.     

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.