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Biomedical subjects

J H Bryan

Publications and source records attributed to J H Bryan.

At least 19 recordsLinked to original sources

A molecular phylogeny of Anopheles annulipes (Diptera: Culicidae) sensu lato: the most species-rich anopheline complex.

The Australasian Annulipes Complex is the most species-rich among Anopheles mosquitoes, with at least 15 sibling species suspected. Members of this complex are the most likely vectors of malaria in the past in southern Australia and are involved in the spread of myxomatosis among rabbits. In this, the first comprehensive molecular study of the Annulipes Complex, 23 ITS2 rDNA variants were detected from collections throughout Australia and Papua New Guinea, including diagnostic variants for the previously identified An. annulipes species A-G. Specimens of each ITS2 variant were sequenced for portions of the mitochondrial COI, COII and nuclear EF-1alpha genes. Partitioned Bayesian and Maximum Parsimony analyses confirmed the monophyly of the Annulipes Complex and revealed at least 17 clades that we designate species A-Q. These species belong to two major clades, one in the north and one mainly in the south, suggesting that climate was a driver of species radiation. We found that 65% (11) of the 17 sibling species recorded here had unique COI sequences, suggesting that DNA barcoding will be useful for diagnosing species within the Annulipes Complex. A comparison of the taxa revealed morphological characters that may be diagnostic for some species. Our results substantially increase the size of the subgenus Cellia in Australasia, and will assist species-level studies of the Annulipes Complex.

Animals↗

Molecular systematics of the Philippine malaria vector Anopheles flavirostris.

Allozyme and molecular sequence data from the malaria vector Anopheles flavirostris (Ludlow) (Diptera: Culicidae) were analysed from 34 sites throughout the Philippines, including the type locality, to test the hypothesis that this taxon is a single panmictic species. A finer-scaled allozyme study, of mainly Luzon samples, revealed no fixed genetic differences in sympatric sites and only low levels of variation. We obtained data from partial sequences for the internal transcribed spacer 2 (ITS2) (483 bp), the third domain (D3) (330 bp) of the 28S ribosomal DNA subunit and cytochrome c oxidase subunit I (COI) of mitochondrial DNA (261 bp). No sequence variation was observed for ITS2, only a one base pair difference was observed between Philippine and Indonesian D3 sequences and An. flavirostris sequences were unique, confirming their diagnostic value for this taxon. Sixteen COI haplotypes were identified, giving 25 parsimony informative sites. Neighbour-Joining, Maximum Parsimony, Maximum Likelihood and Bayesian phylogenetic analysis of COI sequences for An. flavirostris and outgroup taxa revealed strong branch support for the monophyly of An. flavirostris, thus confirming that Philippine populations of this taxon comprise a single separate species within the Minimus Subgroup of the Funestus Group. Variation in the behaviour of An. flavirostris is likely to be intraspecific rather than interspecific in origin.

Animals↗

Population structure of the peridomestic mosquito Ochlerotatus notoscriptus in Australia.

Ochlerotatus notoscriptus (Skuse) (Diptera: Culicidae) is the predominant peridomestic mosquito in Australia where it is the primary vector of dog heartworm, Dirofilaria immitis (Leidy), and a potentially important vector of arboviruses (Barmah Forest, Ross River) with geographical variation of vector competence. Although widespread, Oc. notoscriptus has low dispersal ability, so it may have isolated subpopulations. The identification of gene flow barriers may assist in understanding arbovirus epidemiology and disease risk, and for developing control strategies for this species. We investigated the population structure of Oc. notoscriptus from 17 sites around Australia, using up to 31 putative allozyme loci, 11 of which were polymorphic. We investigated the effect of larval environment and adult morphology on genetic variation. At least five subpopulations were found, four in New South Wales (NSW) and one unique to Darwin. Perth samples appear to be a product of recent colonization from the Australian east coast. For NSW sites, a Mantel test revealed an isolation by distance effect and spatial autocorrelation analysis revealed an area of effective gene flow of 67 km, which is high given the limited dispersal ability of this species. No consistent difference was observed between 'urban' and 'sylvan' habitats, which suggests frequent movement between these sites. However, a finer-scaled habitat study at Darwin revealed small but significant allele frequency differences, including for Gpi. No fixed allozyme differences were detected for sex, size, integument colour or the colour of species-diagnostic pale scales on the scutum. The domestic habit of Oc. notoscriptus and assisted dispersal have helped to homogenize this species geographically but population structure is still detectable on several levels associated with geographical variation of vector competence.

Alleles↗

A morphological study of the Anopheles punctulatus group (Diptera: Culicidae) in the Solomon Islands, with a description of Anopheles (Cellia) irenicus Schmidt, sp.n.

A description of Anopheles (Cellia) irenicus Schmidt, sp.n. (formerly A. farauti No. 7) is provided. This species is one of six recorded from the Solomon Islands within the A. punctulatus group, which contains the major vectors of the causative agents of malaria and lymphatic filariasis in the southwest Pacific. Morphological markers are described for adult females, fourth-instar larvae and pupae that identify most specimens of A. irenicus. Keys are presented to distinguish members of the A. punctulatus group in the Solomon Islands.

Animals↗

Descriptions of the Anopheles (Cellia) farauti complex of sibling species (Diptera: Culicidae) in Australia.

Descriptions of the three sibling species of the Anopheles farauti complex in Australia, A. farauti Laveran (formerly A. farauti No. 1), A. hinesorum Schmidt sp.n. (formerly A. farauti No. 2) and A. torresiensis Schmidt sp.n. (formerly A. farauti No. 3) are provided. These species form a part of the punctulatus group, which contains the major malaria vectors in the southwest Pacific. Morphological markers are described for adult females, fourth instar larvae and pupae which identify most specimens, and are presented in keys.

Animals↗

Ecological distribution of mosquito larvae of the Anopheles punctulatus group on Niolam (Lihir) Island, Papua New Guinea.

We surveyed the larval habitats of members of the Anopheles punctulatus group of mosquitoes on Niolam (Lihir) Island, Papua New Guinea. Identification of this group was undertaken by polymerase chain reaction-restriction fragment length polymorphism analysis of the amplified internal transcribed spacer unit 2 of rDNA, because morphologic separation of member species is unreliable. The most widespread malaria vector species and their most common larval habitats were identified to aid source-reduction programs for malaria control. The most ubiquitous species was An. punctulatus, followed by An. farauti no. 2. then An. farauti s.s. Anopheles punctulatus has increased relative to An. farauti s.l. since the start of development projects on Lihir Island. The most common larval habitats were shallow temporary pools with clay substrate and with plants or floatage. These habitats, mostly encountered alongside poorly drained roads, may be increased by development projects.

Animals↗

Malaria control in central Malaita, Solomon Islands. 1. The use of insecticide-impregnated bed nets.

The present study investigated the use of insecticide-impregnated bed nets by communities in central Malaita, Solomon Islands. Qualitative and quantitative data were collected by: (1) questionnaire administration to 124 care-givers of children aged 0-10 years of age; (2) 20 focus group discussions; (3) two structured observations of bed net re-impregnation, and (4) interviews with key informants. Ninety-four percent of all care-givers had bed nets, but only 62% had sufficient bed nets for all household members. Fifty-two percent used bed nets throughout the year and 70% of care-givers reported that all their children slept under bed nets. Although coastal householders considered malaria and mosquitoes more of a problem than inland householders, overall bed net compliance did not differ. Factors affecting bed net ownership were cost and community expectation of free bed nets. Bed net use was affected by four factors: (1) seasonality (99% used bed nets during the rainy season, 52% used them all year); (2) mosquito nuisance (59% of respondents reported that protection against mosquitoes was the main reason for using a bed net); (3) weather (68% of care-givers would not use a bed net if the weather was hot), and (4) low density of mosquitoes (respondents who used bed nets as protection against mosquito nuisance were more likely not to use bed nets when mosquitoes were few than those who used bed nets for malaria protection (odds ratio (OR), 3.9; 95% confidence interval (CI), 1.4-12.0). Protection against malaria was the main reason children slept under bed nets. Children from households where bed nets were used for malaria protection were more likely to sleep under bed nets than children from households where nets were used as protection from mosquitoes only (OR, 2.7; 95% CI, 1.3-5.9). Other factors that affected children's bed net use were, age (users were significantly younger than non-users; chi(2)=7.9, degrees of freedom=1, P=0.005) and sufficiency of bed nets (OR, 2.0; 95% CI, 1. 3-7.0).

Animals↗

Malaria control in central Malaita, Solomon Islands 2. Local perceptions of the disease and practices for its treatment and prevention.

Government health policy for malaria control in Solomon Islands has three main objectives: (1) early diagnosis and treatment of malaria at a health service; (2) reduction of human-vector contact through widespread use of insecticide-impregnated bed nets; and (3) provision of malaria chemoprophylaxis for pregnant women. Social research was carried out in thirteen villages in central Malaita to determine local attitudes toward malaria and to estimate the level of participation in malaria control activities. Interviews with 124 care-givers who had children 0-10 years of age, 20 focus group discussions and four evening structured observations were research methods used. Antimalarial drugs were the most favoured treatment, and use of traditional medicines and healers were reportedly minimal. Twenty-five percent of respondents reported keeping chloroquine at home and 42% said they would use chloroquine before seeking diagnosis and treatment from a health service. Structured observations suggest that protection against mosquitoes is poor during the evening. Fifty-two percent of respondents reported using fire and 32% said they used bed nets to protect themselves from mosquitoes. Participants had contradictory beliefs on the threat of malaria during pregnancy and the safety of taking chloroquine prophylaxis. Implications of malaria treatment and prevention practices are discussed, and recommendations for improving malaria control are presented.

Animals↗

Shared salinity tolerance invalidates a test for the malaria vector Anopheles farauti s.s. on Guadalcanal, Solomon Islands.

Among the Punctulatus Group of Anopheles mosquitoes (Diptera: Culicidae), first-instar larvae of the medically unimportant freshwater Anopheles farauti species No. 7 survives a seawater tolerance test (SST) that was previously thought to be diagnostic for the saltwater-tolerant malaria vector species, An. farauti Laveran s.s. Salt tolerance in these two closely related isomorphic species appears to be a shared derived character within the Farauti Complex. Failure to differentiate An. farauti s.s. from An. farauti No. 7 will overestimate potential malaria vector numbers and waste limited larval control resources. Use of the SST should therefore be discontinued on Guadalcanal and other techniques such as allozyme electrophoresis used instead.

Animals↗

Shared salinity tolerance invalidates a test for the malaria vector Anopheles farauti s.s. on Guadalcanal, Solomon Islands [corrected].

Among the Punctulatus Group of Anopheles mosquitoes (Diptera: Culicidae), first-instar larvae of the medically unimportant freshwater Anopheles farauti species No. 7 survives a seawater tolerance test (STT) that was previously thought to be diagnostic for the saltwater-tolerant malaria vector species, An. farauti Laveran s.s. Salt tolerance in these two closely related isomorphic species appears to be a shared derived character within the Farauti Complex. Failure to differentiate An. farauti s.s. from An. farauti No.7 will overestimate potential malaria vector numbers and waste limited larval control resources. Use of the STT should therefore be discontinued on Guadalcanal and other techniques such as allozyme electrophoresis used instead [corrected].

Animals↗

Seasonal abundance of Anopheles farauti (Diptera: Culicidae) sibling species in far north Queensland, Australia.

In the Cairns area of far north Queensland, Australia, the seasonal abundance of Anopheles farauti Laveran sibling species was studied at 6 locations, representing 3 habitat types, between August 1995 and September 1997. A total of 45,401 An. farauti s.l. was collected using CO2 + octenol baited CDC light traps, and consisted of 29,565 An. farauti No. 2, 14,214 An. farauti No. 3, and 1,622 An. farauti s.s. The relative abundance of all 3 species differed significantly by season and location. An. farauti No.2 was the dominant species except in Cairns, where An. farauti s.s. was most abundant, and at Ninds Creek, where An. farauti No. 3 predominated. The dominant species at each location was present year round, although peaks in seasonal abundance were observed. An. farauti s.s. populations were highest during the wet season (January-April). In lowland freshwater swamp habitats and 1 brackish location, An. farauti No. 2 was more abundant during the wet season. However, at the highland freshwater swamp habitat, populations of An. farauti No. 2 were highest during the late dry season and early wet season (October-December). There was a significant positive correlation of both temperature and rainfall with An. farauti s.s. and An. farauti No. 2 trap collections. There was a negative correlation between An. farauti No. 3 and temperature, indicating that this species may be more abundant during cool weather. Although there were significant relationships among weather variables and An. farauti s.l. collections, correlation values were generally low, indicating that other factors may contribute to variability among An. farauti sibling species trap collections.

Analysis of Variance↗

Anopheles kochi in Irian Jaya detected by size polymorphism of polymerase chain reaction-amplified internal transcribed spacer unit 2.

Anopheles kochi is reported for the 1st time from New Guinea, probably introduced by aircraft. This Oriental species was originally detected by analysis of polymerase chain reaction-amplified rDNA internal transcribed spacer unit 2 (ITS-2). Identification was confirmed by morphologic examination. Size of ITS-2 is presented for 32 species of Australasian and Oriental anophelines to assist morphologic identifications for distribution and vector studies.

Animals↗

Oviposition preference for freshwater in the coastal malaria vector, Anopheles farauti.

Oviposition preference of the Australasian coastal malaria vector Anopheles farauti s.s. for water of varying salinity was determined in the laboratory to help understand the distribution and control of this species in the field. Numbers of eggs laid showed an inverse relationship with salinity; of 5 NaCl concentrations most eggs were laid in distilled water but some were laid in 3.17% NaCl (the salinity of seawater). The association of An. farauti with coastal areas occurs in spite of an aversion to salt water by ovipositing females. Factors other than salinity must be the primary determinants of distribution. Increasing the salinity of larval habitats will not totally prevent An. farauti from laying eggs. Elimination of this species may not occur unless salinity is kept high enough to prevent complete larval development.

Animals↗

Evolution and systematics of Anopheles: insights from a molecular phylogeny of Australasian mosquitoes.

Relationships among the genus Anopheles and its many sibling species-groups are obscure despite the importance of anophelines as the vectors of human malaria. For the first time, the interrelationships and the origin of Australasian members of the subgenus Cellia are investigated by a cladistic analysis of sequence variation within the mitochondrial cytochrome oxidase subunit II gene. Estimated divergence times between many Australasian and Oriental taxa predate the mid Miocene collision of Australasia and Southeast Asia. Phylogenetic analysis suggests that two-way exchanges with Oriental mosquitoes rather than only immigration may have been a characteristic of anopheline paleobiogeography in Australasia. The Australasian fauna is mostly included in a large clade. The medically important Punctulatus Group is monophyletic and appears derived from Oriental stock. Populations within this group from as far apart as Australia and Vanuatu were in contact in the recent past (i.e., 0.35-2.44 mya), supporting dispersal rather than vicariance explanations. Some support for the monophyly of the Myzomyia, Neomyzomyia, and Pyretophorus Series was found. However, the subgenera Anopheles and Cellia and the Neocellia Series are paraphyletic, but branch support at these taxonomic levels was poor. The COII gene shows promise for questions concerning alpha taxonomy but appears to be of limited use for resolving deeper relationships within the Anopheles.

Amino Acid Sequence↗

Malaria transmission and climate change in Australia.

Although endemic malaria was eradicated from Australia by 1981, the vectors remain and transmission from imported cases still occurs. Climate modelling shows that global warming will enlarge the potential range of the main vector, Anopheles farauti sensu stricto; by the year 2030 it could extend along the Queensland coast to Gladstone, 800 km south of its present limit. Vigilance and a dispassionate assessment of risk are needed to meet this challenge.

Animals↗

DNA probes for the Anopheles punctulatus complex.

Genomic DNA probes were made for two recently identified members of the Anopheles punctulatus complex; Anopheles sp. near punctulatus from Papua New Guinea and Anopheles farauti No. 7 from the Solomon Islands. The probes are species-specific and with the use of 32P labeling sensitive enough so that a squash blot of only a small segment of the mosquito is required for identification. The 119-basepair (bp) probe for An. sp. near punctulatus and the 1,106-bp probe for An. farauti No. 7 have been sequenced in full and the probes have been tested on field collected specimens. These probes now make it possible to distinguish An. sp. near punctulatus and An. farauti No. 7 from the other eight members of the An. punctulatus complex. A pan-species probe was also made from the 18S ribosomal DNA that binds to DNA from all members of the complex. These three probes complete the set required for distinguishing all known members of the An. punctulatus complex by DNA hybridization.

Animals↗

Malaria in Honiara, Solomon Islands: vector studies.

Adult and larval specimens of anopheline mosquitos were collected throughout eastern Honiara during a study into risk factors for malaria illness in adults. Species identification was by morphology, DNA probes and by PCR. Only Anopheles farauti s.s. were identified from part-night landing catches carried out from 1900 to 2200 hours. Most mosquitos attracted to humans were culicines. The majority of anophelines (85%) were captured between 1900 and 2000 hours. An. farauti s.s. larvae were most common but one An. farauti No. 7, and ten An. punctulatus larvae were also collected.

Adult↗