scholarly journals Ecological drivers of genetic connectivity for African malaria vectors Anopheles gambiae and An. arabiensis

2020 ◽  
Vol 10 (1) ◽  
Author(s):  
Elizabeth Hemming-Schroeder ◽  
Daibin Zhong ◽  
Maxwell Machani ◽  
Hoan Nguyen ◽  
Sarah Thong ◽  
...  
Keyword(s):  
Genetic Diversity ◽  
Gene Flow ◽  
Anopheles Gambiae ◽  
Population Genetic ◽  
Sub Saharan Africa ◽  
Tree Cover ◽  
Genetic Connectivity ◽  
Malaria Vectors ◽  
Western Kenya ◽  
Sub Saharan

AbstractAnopheles gambiae and An. arabiensis are major malaria vectors in sub-Saharan Africa. Knowledge of how geographical factors drive the dispersal and gene flow of malaria vectors can help in combatting insecticide resistance spread and planning new vector control interventions. Here, we used a landscape genetics approach to investigate population relatedness and genetic connectivity of An. gambiae and An. arabiensis across Kenya and determined the changes in mosquito population genetic diversity after 20 years of intensive malaria control efforts. We found a significant reduction in genetic diversity in An. gambiae, but not in An. arabiensis as compared to prior to the 20-year period in western Kenya. Significant population structure among populations was found for both species. The most important ecological driver for dispersal and gene flow of An. gambiae and An. arabiensis was tree cover and cropland, respectively. These findings highlight that human induced environmental modifications may enhance genetic connectivity of malaria vectors.

scholarly journals Determination of the discriminating concentration of chlorfenapyr (pyrrole) and Anopheles gambiae sensu lato susceptibility testing in preparation for distribution of Interceptor® G2 insecticide-treated nets

2021 ◽  
Vol 20 (1) ◽  
Author(s):  
Richard M. Oxborough ◽  
Aklilu Seyoum ◽  
Yemane Yihdego ◽  
Joseph Chabi ◽  
Francis Wat’senga ◽  
...  
Keyword(s):  
Anopheles Gambiae ◽  
Large Scale ◽  
Resistance Mechanisms ◽  
Probit Analysis ◽  
Sub Saharan Africa ◽  
Malaria Vectors ◽  
Insecticide Treated Nets ◽  
Distributed Testing ◽  
The Difference ◽  
Sub Saharan

Abstract Background Following agricultural use and large-scale distribution of insecticide-treated nets (ITNs), malaria vector resistance to pyrethroids is widespread in sub-Saharan Africa. Interceptor® G2 is a new dual active ingredient (AI) ITN treated with alpha-cypermethrin and chlorfenapyr for the control of pyrethroid-resistant malaria vectors. In anticipation of these new nets being more widely distributed, testing was conducted to develop a chlorfenapyr susceptibility bioassay protocol and gather susceptibility information. Methods Bottle bioassay tests were conducted using five concentrations of chlorfenapyr at 12.5, 25, 50, 100, and 200 µg AI/bottle in 10 countries in sub-Saharan Africa using 13,639 wild-collected Anopheles gambiae sensu lato (s.l.) (56 vector populations per dose) and 4,494 pyrethroid-susceptible insectary mosquitoes from 8 colonized strains. In parallel, susceptibility tests were conducted using a provisional discriminating concentration of 100 µg AI/bottle in 16 countries using 23,422 wild-collected, pyrethroid-resistant An. gambiae s.l. (259 vector populations). Exposure time was 60 min, with mortality recorded at 24, 48 and 72 h after exposure. Results Median mortality rates (up to 72 h after exposure) of insectary colony mosquitoes was 100% at all five concentrations tested, but the lowest dose to kill all mosquitoes tested was 50 µg AI/bottle. The median 72-h mortality of wild An. gambiae s.l. in 10 countries was 71.5, 90.5, 96.5, 100, and 100% at concentrations of 12.5, 25, 50, 100, and 200 µg AI/bottle, respectively. Log-probit analysis of the five concentrations tested determined that the LC95 of wild An. gambiae s.l. was 67.9 µg AI/bottle (95% CI: 48.8–119.5). The discriminating concentration of 203.8 µg AI/bottle (95% CI: 146–359) was calculated by multiplying the LC95 by three. However, the difference in mortality between 100 and 200 µg AI/bottle was minimal and large-scale testing using 100 µg AI/bottle with wild An. gambiae s.l. in 16 countries showed that this concentration was generally suitable, with a median mortality rate of 100% at 72 h. Conclusions This study determined that 100 or 200 µg AI/bottle chlorfenapyr in bottle bioassays are suitable discriminating concentrations for monitoring susceptibility of wild An. gambiae s.l., using mortality recorded up to 72 h. Testing in 16 countries in sub-Saharan Africa demonstrated vector susceptibility to chlorfenapyr, including mosquitoes with multiple resistance mechanisms to pyrethroids.

scholarly journals Reticulate speciation and adaptive introgression in the Anopheles gambiae species complex

2014 ◽  
Author(s):  
Jacob Crawford ◽  
Michelle M. Riehle ◽  
Wamdaogo M. Guelbeogo ◽  
Awa Gneme ◽  
N'fale Sagnon ◽  
...  
Keyword(s):  
Gene Flow ◽  
Reproductive Isolation ◽  
Anopheles Gambiae ◽  
X Chromosome ◽  
Species Complex ◽  
Sub Saharan Africa ◽  
Contemporary Gene Flow ◽  
Speciation Continuum ◽  
Primary Vector ◽  
Sub Saharan

Anopheles gambiae, the primary vector of human malaria in sub-Saharan Africa, exists as a series of ecologically specialized subgroups that are phylogenetically nested within the Anopheles gambiae species complex. These species and subgroups exhibit varying degrees of reproductive isolation, sometimes recognized as distinct subspecies. We have sequenced 32 complete genomes from field-captured individuals of Anopheles gambiae, Anopheles gambiae M form (recently named A. coluzzii), sister species A. arabiensis, and the recently discovered ?GOUNDRY? subgroup of A. gambiae that is highly susceptible to Plasmodium. Amidst a backdrop of strong reproductive isolation and adaptive differentiation, we find evidence for introgression of autosomal chromosomal regions among species and subgroups, some of which have facilitated adaptation. The X chromosome, however, is strongly differentiated among all species and subgroups, pointing to a disproportionately large effect of X chromosome genes in driving speciation among anophelines. Strikingly, we find that autosomal introgression has occurred from contemporary hybridization among A. gambiae and A. arabiensis despite strong divergence (~5? higher than autosomal divergence) and isolation on the X chromosome. We find a large region of the X chromosome that has swept to fixation in the GOUNDRY subgroup within the last 100 years, which may be an inversion that serves as a partial barrier to contemporary gene flow. We show that speciation with gene flow results in genomic mosaicism of divergence and introgression. Such a reticulate gene pool connecting vector species and subgroups across the speciation continuum has important implications for malaria control efforts.

scholarly journals Spatial modelling for population replacement of mosquito vectors at continental scale

2021 ◽  
Author(s):  
Nicholas J. Beeton ◽  
Andrew Wilkins ◽  
Adrien Ickowicz ◽  
Keith R. Hayes ◽  
Geoffrey R. Hosack
Keyword(s):  
Anopheles Gambiae ◽  
Control Strategies ◽  
Drive System ◽  
Sub Saharan Africa ◽  
Malaria Vectors ◽  
Anopheles Coluzzii ◽  
Gene Drive ◽  
Continental Scale ◽  
Wide Range ◽  
Sub Saharan

AbstractMalaria is one of the deadliest vector-borne diseases in the world. Researchers are developing new genetic and conventional vector control strategies to attempt to limit its burden. To be deployed responsibly and successfully, proposed novel control strategies require detailed safety assessment. Anopheles gambiae sensu stricto (s.s.) and Anopheles coluzzii, two closely related subspecies within the species complex Anopheles gambiae sensu lato (s.l.), are among the dominant malaria vectors in sub-Saharan Africa. These two subspecies readily hybridise and compete in the wild and are also known to have distinct niches, each with spatially and temporally varying carrying capacities driven by precipitation and land use factors.We model the spread and persistence of a population-modifying gene drive system in these subspecies across sub-Saharan Africa, by simulating introductions of genetically modified mosquitoes across the African mainland as well as on some offshore islands. We explore transmission of the gene drive between the subspecies, different hybridisation mechanisms, the effects of both local dispersal and potential wind-aided migration to the spread, and the development of resistance to the gene drive. We find that given best current available knowledge on the subspecies’ life histories, an introduced gene drive system with typical characteristics can plausibly spread from even distant offshore islands to the African mainland with the aid of wind-driven migration, with resistance taking over within a decade. Our model demonstrates a range of realistic dynamics including the effect of prevailing wind on spread and spatio-temporally varying carrying capacities for subspecies. We thus show both the plausibility and importance of accounting for a wide range of mechanisms from regional to continental scales.

scholarly journals Patterns of Mitochondrial Variation Within and Between African Malaria Vectors, Anopheles gambiae and An. arabiensis, Suggest Extensive Gene Flow

Genetics ◽  
1997 ◽  
Vol 147 (4) ◽  
pp. 1817-1828 ◽  
Author(s):  
Nora J Besansky ◽  
Tovi Lehmann ◽  
G Thomas Fahey ◽  
Didier Fontenille ◽  
Lawrence E O Braack ◽  
...  
Keyword(s):  
Gene Flow ◽  
Anopheles Gambiae ◽  
Genetic Divergence ◽  
Isolation By Distance ◽  
Haplotype Diversity ◽  
Introgressive Hybridization ◽  
Sub Saharan Africa ◽  
Malaria Vectors ◽  
Amino Acid Difference ◽  
Mtdna Sequence

Abstract Anopheles gambiae and An. arabiensis are mosquito species responsible for most malaria transmission in sub-Saharan Africa. They are also closely related sibling species that share chromosomal and molecular polymorphisms as a consequence of incomplete lineage sorting or introgressive hybridization. To help resolve these processes, this study examined the partitioning of mtDNA sequence variation within and between species across Africa, from both population genetic and phylogeographic perspectives. Based on partial gene sequences from the cytochrome b, ND1 and ND5 genes, haplotype diversity was high but sequences were very closely related. Within species, little or no population subdivision was detected, and there was no evidence for isolation by distance. Between species, there were no fixed nucleotide differences, a high proportion of shared polymorphisms, and eight haplotypes in common over distances as great as 6000 km. Only one of 16 shared polymorphisms led to an amino acid difference, and there was no compelling evidence for nonneutral variation. Parsimony networks constructed of haplotypes from both species revealed no correspondence of haplotype with either geography or taxonomy. This trend of low intraspecific genetic divergence is consistent with evidence from allozyme and microsatellite data and is interpreted in terms of both extensive gene flow and recent range expansion from relatively large, stable populations. We argue that retention of ancestral polymorphisms is a plausible but insufficient explanation for low interspecific genetic divergence, and that extensive hybridization is a contributing factor.

scholarly journals Insecticide resistance status of indoor and outdoor resting malaria vectors in a highland and lowland site in Western Kenya

2020 ◽  
Author(s):  
Kevin O. Owuor ◽  
Maxwell G. Machani ◽  
Wolfgang R. Mukabana ◽  
Stephen Munga ◽  
Guiyun Yan ◽  
...  
Keyword(s):  
Insecticide Resistance ◽  
Anopheles Gambiae ◽  
Sub Saharan Africa ◽  
Malaria Vectors ◽  
Moderate Intensity ◽  
Monooxygenase Activity ◽  
Western Kenya ◽  
Malaria Mosquitoes ◽  
Outdoor Resting ◽  
Indoor And Outdoor

AbstractBackgroundLong Lasting Insecticidal Nets (LLINs) and indoor residual spraying (IRS) represent powerful tools for controlling malaria vectors in sub-Saharan Africa. The success of these interventions relies on their capability to inhibit indoor feeding and resting of malaria mosquitoes. This study sought to understand the interaction of insecticide resistance with indoor and outdoor resting behavioral responses of malaria vectors from Western Kenya.MethodsThe status of insecticide resistance among indoor and outdoor resting anopheline mosquitoes was compared in Anopheles mosquitoes collected from Kisumu and Bungoma counties in Western Kenya. The level and intensity of resistance were measured using WHO-tube and CDC-bottle bioassays, respectively. The synergist piperonyl butoxide (PBO) was used to determine if metabolic activity (monooxygenase enzymes) explained the resistance observed. The mutations at the voltage-gated sodium channel (Vgsc) gene and Ace 1 gene were characterized using PCR methods. Microplate assays were used to measure levels of detoxification enzymes if present.ResultsA total of 1094 samples were discriminated within Anopheles gambiae s.l. and 289 within An. funestus s.l. In Kisian (Kisumu county), the dominant species was Anopheles arabiensis 75.2% (391/520) while in Kimaeti (Bungoma county) collections the dominant sibling species was Anopheles gambiae s.s 96.5% (554/574). The An. funestus s.l samples analysed were all An. funestus s.s from both sites. Pyrethroid resistance of An.gambiae s.l F1 progeny was observed in all sites. Lower mortality was observed against deltamethrin for the progeny of indoor resting mosquitoes compared to outdoor resting mosquitoes (Mortality rate: 37% vs 51%, P=0.044). The intensity assays showed moderate-intensity resistance to deltamethrin in the progeny of mosquitoes collected from indoors and outdoors in both study sites. In Kisian, the frequency of vgsc-L1014S and vgsc-L1014F mutation was 0.14 and 0.19 respectively in indoor resting malaria mosquitoes while those of the outdoor resting mosquitoes were 0.12 and 0.12 respectively. The ace 1 mutation was present in higher frequency in the F1 of mosquitoes resting indoors (0.23) compared to those of mosquitoes resting outdoors (0.12). In Kimaeti, the frequencies of vgsc-L1014S and vgsc-L1014F were 0.75 and 0.05 respectively for the F1 of mosquitoes collected indoors whereas those of outdoor resting ones were 0.67 and 0.03 respectively. The ace 1 G119S mutation was present in progeny of mosquitoes from Kimaeti resting indoors (0.05) whereas it was absent in those resting outdoors. Monooxygenase activity was elevated by 1.83 folds in Kisian and by 1.33 folds in Kimaeti for mosquitoes resting indoors than those resting outdoors respectively.ConclusionThe study recorded high phenotypic, metabolic and genotypic insecticide resistance in indoor resting populations of malaria vectors compared to their outdoor resting counterparts. The indication of moderate resistance intensity for the indoor resting mosquitoes is alarming as it could have an operational impact on the efficacy of the existing pyrethroid based vector control tools. The use of synergist (PBO) in LLINs may be a better alternative for widespread use in these regions recording high insecticide resistance.

scholarly journals Insecticide resistance status of indoor and outdoor resting malaria vectors in a highland and lowland site in Western Kenya

PLoS ONE ◽  
2021 ◽  
Vol 16 (3) ◽  
pp. e0240771
Author(s):  
Kevin O. Owuor ◽  
Maxwell G. Machani ◽  
Wolfgang R. Mukabana ◽  
Stephen O. Munga ◽  
Guiyun Yan ◽  
...  
Keyword(s):  
Insecticide Resistance ◽  
Anopheles Gambiae ◽  
Sub Saharan Africa ◽  
Malaria Vectors ◽  
Moderate Intensity ◽  
Monooxygenase Activity ◽  
Western Kenya ◽  
Malaria Mosquitoes ◽  
Outdoor Resting ◽  
Indoor And Outdoor

Background Long Lasting Insecticidal Nets (LLINs) and indoor residual spraying (IRS) represent powerful tools for controlling malaria vectors in sub-Saharan Africa. The success of these interventions relies on their capability to inhibit indoor feeding and resting of malaria mosquitoes. This study sought to understand the interaction of insecticide resistance with indoor and outdoor resting behavioral responses of malaria vectors from Western Kenya. Methods The status of insecticide resistance among indoor and outdoor resting anopheline mosquitoes was compared in Anopheles mosquitoes collected from Kisumu and Bungoma counties in Western Kenya. The level and intensity of resistance were measured using WHO-tube and CDC-bottle bioassays, respectively. The synergist piperonyl butoxide (PBO) was used to determine if metabolic activity (monooxygenase enzymes) explained the resistance observed. The mutations at the voltage-gated sodium channel (Vgsc) gene and Ace 1 gene were characterized using PCR methods. Microplate assays were used to measure levels of detoxification enzymes if present. Results A total of 1094 samples were discriminated within Anopheles gambiae s.l. and 289 within An. funestus s.l. In Kisian (Kisumu county), the dominant species was Anopheles arabiensis 75.2% (391/520) while in Kimaeti (Bungoma county) collections the dominant sibling species was Anopheles gambiae s.s 96.5% (554/574). The An. funestus s.l samples analysed were all An. funestus s.s from both sites. Pyrethroid resistance of An.gambiae s.l F1 progeny was observed in all sites. Lower mortality was observed against deltamethrin for the progeny of indoor resting mosquitoes compared to outdoor resting mosquitoes (Mortality rate: 37% vs 51%, P = 0.044). The intensity assays showed moderate-intensity resistance to deltamethrin in the progeny of mosquitoes collected from indoors and outdoors in both study sites. In Kisian, the frequency of vgsc-L1014S and vgsc-L1014F mutation was 0.14 and 0.19 respectively in indoor resting malaria mosquitoes while those of the outdoor resting mosquitoes were 0.12 and 0.12 respectively. The ace 1 mutation was present in higher frequency in the F1 of mosquitoes resting indoors (0.23) compared to those of mosquitoes resting outdoors (0.12). In Kimaeti, the frequencies of vgsc-L1014S and vgsc-L1014F were 0.75 and 0.05 respectively for the F1 of mosquitoes collected indoors whereas those of outdoor resting ones were 0.67 and 0.03 respectively. The ace 1 G119S mutation was present in progeny of mosquitoes from Kimaeti resting indoors (0.05) whereas it was absent in those resting outdoors. Monooxygenase activity was elevated by 1.83 folds in Kisian and by 1.33 folds in Kimaeti for mosquitoes resting indoors than those resting outdoors respectively. Conclusion The study recorded high phenotypic, metabolic and genotypic insecticide resistance in indoor resting populations of malaria vectors compared to their outdoor resting counterparts. The indication of moderate resistance intensity for the indoor resting mosquitoes is alarming as it could have an operational impact on the efficacy of the existing pyrethroid based vector control tools. The use of synergist (PBO) in LLINs may be a better alternative for widespread use in these regions recording high insecticide resistance.

scholarly journals A pyrethroïd-treated bed net increases host attractiveness for Anopheles gambiae s.s. carrying the kdr allele in a dual-choice olfactometer

2016 ◽  
Author(s):  
Angélique Porciani ◽  
Malal Diop ◽  
Nicolas Moiroux ◽  
Tatiana Kadoke-Lambi ◽  
Anna Cohuet ◽  
...  
Keyword(s):  
Insecticide Resistance ◽  
Anopheles Gambiae ◽  
Sub Saharan Africa ◽  
Malaria Vectors ◽  
Malaria Vector Control ◽  
Anopheles Gambiae S.S ◽  
Host Seeking ◽  
Seeking Behavior ◽  
Sub Saharan ◽  
The Impact

AbstractThe use of long lasting insecticide nets (LLINs) treated with pyrethroïd is known for its major contribution in malaria control. However, LLINs are suspected to induce behavioral changes in malaria vectors, which may in turn drastically affect their efficacy against Plasmodium sp. transmission. In sub Saharan Africa, where malaria imposes the heaviest burden, the main malaria vectors are widely resistant to pyrethroïds, the insecticide family used on LLINs, which also threatens LLIN efficiency. There is therefore a crucial need for deciphering how insecticide-impregnated materials might affect the host-seeking behavior of malaria vectors in regards to insecticide resistance. In this study, we explored the impact of permethrin-impregnated net on the host attractiveness for Anopheles gambiae mosquitoes, either susceptible to insecticides, or carrying the insecticide resistance conferring allele kdr. Groups of female mosquitoes were released in a dual-choice olfactometer and their movements towards an attractive odor source (a rabbit) protected by insecticide-treated (ITN) or untreated nets (UTN) were monitored. Kdr homozygous mosquitoes, resistant to insecticides, were more attracted by a host behind an ITN than an UTN, while the presence of insecticide on the net did not affect the choice of susceptible mosquitoes. These results suggest that permethrin-impregnated net is detectable by malaria vectors and that the kdr mutation impacts their response to a LLIN protected host. We discuss the implication of these results for malaria vector control.

scholarly journals Resting Behavior of Malaria Vectors in Ghana and Its Implication on Vector Control.

2022 ◽  
Author(s):  
Akua Obeng Forson ◽  
Isaac A. Hinne ◽  
Shittu B. Dhikrullahi ◽  
Isaac Kwame Sr ◽  
Abdul Rahim Mohammed ◽  
...  
Keyword(s):  
Vector Control ◽  
Anopheles Gambiae ◽  
Blood Meal ◽  
Indoor Residual Spraying ◽  
Sub Saharan Africa ◽  
Malaria Vectors ◽  
Resistance Mutations ◽  
Sub Saharan ◽  
Outdoor Resting ◽  
Resting Behavior

Abstract Background: In Sub-Saharan Africa, there is widespread use of long-lasting insecticidal nets (LLINs) and Indoor residual spraying (IRS) to help control the density of malaria vectors and decrease the incidence of malaria in communities. An understanding of the interactions between increased insecticide use and resting behaviour patterns of malaria mosquitoes is important for an effective vector control programme. This study was carried out to investigate the resting behavior, host preference and infection with Plasmodium falciparum of malaria vectors in Ghana in the context of increasing insecticide resistance in malaria vectors in sub-saharan Africa.Methods: Indoor and outdoor resting Anopheline mosquitoes were sampled during the dry and rainy seasons in five sites that were in 3 ecological landscapes [Sahel savannah (Kpalsogou, Pagaza, Libga), Coastal savannah (Anyakpor) and Forest (Konongo) zones] using pyrethrum spray catches (PSC), mechanical aspiration (Prokopack) for indoor collections, pit shelter and Prokopack for outdoor collections. PCR based molecular diagnostics were used to determine mosquito speciation, genotype for knockdown resistance mutations (L1014S and L1014F), G119S Ace-1 mutation, specific host blood meal origins and sporozoite infection in field collected mosquitoes.Results: Anopheles gambiae s. l. was the predominant species (89.95%, n = 1,718), followed by An. rufipes (8.48%, n=162), and An. funestus s. l. (1.57%, n = 30). Sibling species of the Anopheles gambiae revealed An. coluzzii accounted for 63% (95% CI: 57.10 – 68.91), followed by An. gambiae s. s [27% (95% CI: 21.66 – 32.55)], and An. arabiensis [9% (95% CI: 6.22 – 13.57)]. The mean resting density of An. gambiae s. l. was higher outdoors (79.63%; 1,368/1,718) than indoors (20.37%; 350/1,718) (z = -4.815, p< 0.0001). The kdr west L1014F and the Ace-1 mutations were highest in indoor resting An. coluzzii and An. gambiae in the sahel-savannah sites compared to the forest and coastal savannah sites. Overall, the blood meal analyses revealed a large proportion of the malaria vectors preferred feeding on humans (70.2 %) than animals (29.8%) in all sites. The sporozoite rates was only detected in indoor resting An. coluzzii from the sahel savannah (5.0%) and forest (2.5%) zones.Conclusion: The study reports high outdoor resting densities of An. gambiae and An. coluzzii with high kdr west mutation frequencies, and persistence of malaria transmission indoors despite the use of LLINs and IRS. Continuous monitoring of changes in resting behavior of mosquitoes and implementation of complementary malaria control interventions are needed to target outdoor resting Anopheles mosquitoes in Ghana.

scholarly journals Evolutionary history and genetic connectivity across highly fragmented populations of an endangered daisy

Heredity ◽  
2021 ◽  
Author(s):  
Yael S. Rodger ◽  
Alexandra Pavlova ◽  
Steve Sinclair ◽  
Melinda Pickup ◽  
Paul Sunnucks
Keyword(s):  
Genetic Diversity ◽  
Gene Flow ◽  
Genetic Differentiation ◽  
Evolutionary History ◽  
Livestock Grazing ◽  
Genetic Connectivity ◽  
Evolutionary Divergence ◽  
Common Component ◽  
A Genome ◽  
Eastern Australia

AbstractConservation management can be aided by knowledge of genetic diversity and evolutionary history, so that ecological and evolutionary processes can be preserved. The Button Wrinklewort daisy (Rutidosis leptorrhynchoides) was a common component of grassy ecosystems in south-eastern Australia. It is now endangered due to extensive habitat loss and the impacts of livestock grazing, and is currently restricted to a few small populations in two regions >500 km apart, one in Victoria, the other in the Australian Capital Territory and nearby New South Wales (ACT/NSW). Using a genome-wide SNP dataset, we assessed patterns of genetic structure and genetic differentiation of 12 natural diploid populations. We estimated intrapopulation genetic diversity to scope sources for genetic management. Bayesian clustering and principal coordinate analyses showed strong population genetic differentiation between the two regions, and substantial substructure within ACT/NSW. A coalescent tree-building approach implemented in SNAPP indicated evolutionary divergence between the two distant regions. Among the populations screened, the last two known remaining Victorian populations had the highest genetic diversity, despite having among the lowest recent census sizes. A maximum likelihood population tree method implemented in TreeMix suggested little or no recent gene flow except potentially between very close neighbours. Populations that were more genetically distinctive had lower genetic diversity, suggesting that drift in isolation is likely driving population differentiation though loss of diversity, hence re-establishing gene flow among them is desirable. These results provide background knowledge for evidence-based conservation and support genetic rescue within and between regions to elevate genetic diversity and alleviate inbreeding.

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