South Australian Populations Research Articles

Floral visitors, pollinators and floral rewards of the Australian dioecious arid zone shrub Pimelea microcephala subsp. microcephala

ABSTRACT Entomophilous dioecious plants may present opportunities to conserve insect populations as they often possess flowers accessible to a diversity of visitors and potential pollinators. Additionally, flowers of each sex can vary in the nutrients they provide. Two South Australian populations of the largely dioecious arid zone shrub Pimelea microcephala subsp. microcephala (Thymelaeaceae) were investigated to determine the conservation utility of the species to conserve insect populations by assessing floral visitor and pollinator diversity and the nutritional quality of pollen and nectar. Fifty-five visitors representing species in six insect orders were recorded at Baron Station, while 49 visitors from three insect orders were recorded at Clements Gap. Thirty-nine of the observed visitors were captured, of which two species of Hymenoptera, two of Diptera and one of Hemiptera were carrying pollen and thus potential pollinators. Visitation rates were higher for male than female plants at Clements Gap, but the reverse at Baron Station. Pollen from male plants had low protein content (2.5% dried weight (DW)) compared to co-flowering hermaphroditic angiosperms at the sites (5.3–14.0% DW). In contrast, floral extracts from male plants had at least 2× greater cumulative sugar concentrations than those of females at the same site, but female extracts had at least 4× higher protein content than male ones. As a food resource plant, P. microcephala subsp. microcephala may represent an important plant species for revegetation efforts that support the conservation of insect diversity in arid regions and throughout its broad range.

The

Crinum flaccidum Herb. is widespread across southern and eastern Australia, but suffers from taxonomic uncertainty. This ambiguity is in part due to widespread hybridisation in the genus, but also morphological variability within and among populations. Morphological and molecular analyses of the complex using 24 morphological characters and 59 chloroplast coding and non-coding regions (~50 000 bp) provided support for the separation of C. flaccidum from C. luteolum Traub & L.S.Hannibal ex Traub, with the latter representing populations from the Flinders Ranges and Lake Eyre Basin. Within Crinum flaccidum, there was greater inferred genetic structure at the population level for New South Wales flood plain accessions, compared with South Australian populations from along the Murray River. The greater structure of the South Australian populations is theorised to be attributed to lower seed-dispersal rates leading to lower gene flow.

The genomic structure of isolation across breed, country and strain for important South African and Australian sheep populations

BackgroundSouth Africa and Australia shares multiple important sheep breeds. For some of these breeds, genomic breeding values are provided to breeders in Australia, but not yet in South Africa. Combining genomic resources could facilitate development for across country selection, but the influence of population structures could be important to the compatability of genomic data from varying origins. The genetic structure within and across breeds, countries and strains was evaluated in this study by population genomic parameters derived from SNP-marker data. Populations were first analysed by breed and country of origin and then by subpopulations of South African and Australian Merinos.ResultsMean estimated relatedness according to the genomic relationship matrix varied by breed (-0.11 to 0.16) and bloodline (-0.08 to 0.06) groups and depended on co-ancestry as well as recent genetic links. Measures of divergence across bloodlines (FST: 0.04–0.12) were sometimes more distant than across some breeds (FST: 0.05–0.24), but the divergence of common breeds from their across-country equivalents was weak (FST: 0.01–0.04). According to mean relatedness, FST, PCA and Admixture, the Australian Ultrafine line was better connected to the SA Cradock Fine Wool flock than with other AUS bloodlines. Levels of linkage disequilibrium (LD) between adjacent markers was generally low, but also varied across breeds (r2: 0.14–0.22) as well as bloodlines (r2: 0.15–0.19). Patterns of LD decay was also unique to breeds, but bloodlines differed only at the absolute level. Estimates of effective population size (Ne) showed genetic diversity to be high for the majority of breeds (Ne: 128–418) but also for bloodlines (Ne: 137–369).ConclusionsThis study reinforced the genetic complexity and diversity of important sheep breeds, especially the Merino breed. The results also showed that implications of isolation can be highly variable and extended beyond breed structures. However, knowledge of useful links across these population substructures allows for a fine-tuned approach in the combination of genomic resources. Isolation across country rarely proved restricting compared to other structures considered. Consequently, research into the accuracy of across-country genomic prediction is recommended.

Predicting the past, present and future distributions of an endangered marsupial in a semi‐arid environment

AbstractGlobally, the impacts of anthropogenic climate change can displace species into more favourable climates. Semi‐arid desert specialists, such as the sandhill dunnart, Sminthopsis psammophila, are typically susceptible to rainfall deficits, wildfires and extreme temperatures caused by anthropogenic climate change. We first used maximum entropy (MaxEnt) species distribution models (SDMs) to predict the current distribution of S. psammophila. Between 2016 and 2018, we ground validated the model’s predictions throughout Western Australia, confirming S. psammophila in 18 locations in which it was predicted to occur. The predicted distribution of S. psammophila appears mostly constrained to within its known range. However, S. psammophila was verified 150 km north of its range in Western Australia and connectivity between the South Australian populations was correctly predicted. In 2019, we used updated occurrence data to project SDMs for S. psammophila during the mid‐Holocene, present day and under two future representative concentration pathways (RCPs) of RCP 4.5 (an optimistic emissions scenario) and RCP 8.5 (“business as usual”) for 2050 and 2070. By 2050 (RCP 8.5), almost all Western Australian Great Victoria Desert (WAGVD) habitat is predicted to be unsuitable for S. psammophila. By 2070 (RCP 8.5), the climates of the WAGVD and Yellabinna Regional Reserve populations are predicted to become unsuitable, and the species’ geographical range is predicted to contract in Australia by 80%. However, the 2070 (RCP 4.5) scenario predicts that this contraction could be halved. As a sandy desert specialist, the distribution of S. psammophila is geographically limited at its southern bounds due to the cessation of suitable spinifex (Triodia spp.) habitats, and so further extension of the range southwards is not possible. Sympatric desert species may be similarly affected, and we suggest that SDMs will be a useful tool in helping to predict the effects of climate change on their distributions.

Identification and Prevalence of Phascolarctid Gammaherpesvirus Types 1 and 2 in South Australian Koala Populations.

To determine Phascolarctid gammaherpesviruses (PhaHV) infection in South Australian koala populations, 80 oropharyngeal swabs from wild-caught and 87 oropharyngeal spleen samples and swabs from euthanased koalas were tested using two specific PCR assays developed to detect PhaHV-1 and PhaHV-2. In wild-caught koalas, active shedding of PhaHV was determined by positive oropharyngeal samples in 72.5% (58/80) of animals, of which 44.8% (26/58) had PhaHV-1, 20.7% (12/58) PhaHV-2 and 34.5% (20/58) both viral subtypes. In the euthanased koalas, systemic infection was determined by positive PCR in spleen samples and found in 72.4% (63/87) of koalas. Active shedding was determined by positive oropharyngeal results and found in 54.0% (47/87) of koalas. Koalas infected and actively shedding PhaHV-1 alone, PhaHV-2 alone or shedding both viral subtypes were 48.9% (23/47), 14.9% (7/47) and 36.2% (17/47), respectively. Only 45.9% (40/87) were not actively shedding, of which 40.0% (16/40) of these had systemic infections. Both wild-caught and euthanased koalas actively shedding PhaHV-2 were significantly more likely to be actively shedding both viral subtypes. Active shedding of PhaHV-2 had a significant negative correlation with BCS in the euthanased cohort, and active shedding of PhaHV-1 had a significant positive relationship with age in both wild-caught and euthanased cohorts.

Spotlight on the April 14 issue

#### Notable in Neurology this week This issue features an article that investigates whether in vivo glycolysis-related glucose metabolism and electron transport chain-related mitochondrial activity are regionally different in the brains of patients with Alzheimer disease; another determines which clinical and demographic factors predict 30-day hospital readmissions. A featured article explores whether amyotrophic lateral sclerosis is a multistep process across South Korean, Japanese, and Australian populations. ### Better stress coping associated with lower tau in amyloid-positive cognitively unimpaired older adults Resilience to stress was associated with lower tau deposition in areas …

Prevalence and clinical significance of koala retrovirus in two South Australian koala (Phascolarctos cinereus) populations.

Koala retrovirus (KoRV-A) is 100 % prevalent in northern Australian (Queensland and New South Wales) koala populations, where KoRV-B has been associated with Chlamydia pecorum disease and the development of lymphosarcoma. In southern populations (Victoria and South Australia), KoRV-A is less prevalent and KoRV-B has not been detected in Victoria, while the current prevalence in South Australian populations is unknown but is thought to be low. This study aimed to determine (i) the prevalence of KoRV in the two largest South Australian koala populations [Kangaroo Island (KI) and Mount Lofty Ranges (MLR)], (ii) KoRV subtype and (iii) if an association between KoRV and C. pecorum exists. Wild koalas were sampled in KI ( n =170) between 2014 and 2017 and in MLR ( n =75) in 2016. Clinical examinations were performed, with blood collected for KoRV detection and typing by PCR. KoRV prevalence was 42.4 % [72/170, 95 % confidence interval (CI): 34.9-49.8 %] in KI and 65.3 % (49/75, 95 % CI: 54.6-76.1 %) in MLR. Only KoRV-A, and not KoRV-B, was detected in both populations. In MLR, there was no statistical association between KoRV and C. pecorum infection (P =0.740), or KoRV and C. pecorum disease status ( P=0.274), although KoRV-infected koalas were more likely to present with overt C. pecorum disease than subclinical infection (odds ratio: 3.15, 95 % CI: 0.91-5.39). KoRV-A is a prevalent pathogen in wild South Australian koala populations. Future studies should continue to investigate KoRV and C. pecorum associations, as the relationship is likely to be complex and to differ between the northern and southern populations.

Chlamydia pecorum prevalence in South Australian koala (Phascolarctos cinereus) populations: Identification and modelling of a population free from infection

Chlamydia pecorum is an established and prevalent infection that produces severe clinical disease in many koala populations, contributing to dramatic population declines. In wild South Australian koala populations, C. pecorum occurrence and distribution is unknown. Here, C. pecorum-specific real-time quantitative PCR (qPCR) was applied to ocular and urogenital swabs from targeted surveys of wild koalas from the mainland Mount Lofty Ranges (MLR) (n = 75) and Kangaroo Island (KI) (n = 170) populations. Historical data from 13,081 KI koalas (1997–2018) provided additional evidence for assessing the absence of C. pecorum infection. In the MLR population, 46.7% (CI: 35.1–58.6%) of koalas were C. pecorum positive by qPCR but only 4% had grade 3 clinical disease. MLR koala fertility was significantly reduced by C. pecorum infection; all reproductively active females (n = 16) were C. pecorum negative, whereas 85.2% of inactive females (n = 23) were positive (P < 0.001). KI koalas were C. pecorum negative and the population was demonstrated to be free of C. pecorum infection with 95% confidence. C. pecorum is a real threat for the sustainability of the koala and KI is possibly the last isolated, large C. pecorum-free population remaining in Australia. These koalas could provide a safeguard against this serious disease threat to an iconic Australian species.

Panel of three loop-mediated isothermal amplification assays differentiates Rathayibacter toxicus populations RT-I, RT-II, RT-III, RT-IV and RT-V

Rathayibacter toxicus is a nematode-vectored (Anguina sp.) plant-pathogenic (Poaceae; grasses) bacterium that secretes a toxin, which can cause fatalities in livestock and horses that ingest diseased plant material. The lethal toxicoses associated with R. toxicus are annual ryegrass toxicity and flood plain staggers. Currently, this bacterium is confined to certain areas of Australia and South Africa. Previous studies have shown that populations from Western Australia (RT-III) are genetically distinct from populations from South Australia (RT-II). A recent study discovered a newly emerged population in South Australia (RT-I), distinct from both Western and South Australian populations studied previously. In this study, a six-gene MLST delineated five distinct genetic populations. Phenotypic significance of these different populations is not yet known. A set of molecular-based assays for the specific detection of R. toxicus has been developed. Described here are loop-mediated isothermal amplification (LAMP) assays for rapid identification and differentiation of R. toxicus populations in the field or laboratory. These assays can be used subsequent to an R. toxicus-specific field assay, allowing on-site determination of population type. This will allow informed field sampling to collect populations of interest, as well as track the movement of populations to new areas.

Microsatellite Analysis and Urediniospore Dispersal Simulations Support the Movement of Puccinia graminis f. sp. tritici from Southern Africa to Australia.

The Australian wheat stem rust (Puccinia graminis f. sp. tritici) population was shaped by the introduction of four exotic incursions into the country. It was previously hypothesized that at least two of these (races 326-1,2,3,5,6 and 194-1,2,3,5,6 first detected in 1969) had an African origin and moved across the Indian Ocean to Australia on high-altitude winds. We provide strong supportive evidence for this hypothesis by combining genetic analyses and complex atmospheric dispersion modeling. Genetic analysis of 29 Australian and South African P. graminis f. sp. tritici races using microsatellite markers confirmed the close genetic relationship between the South African and Australian populations, thereby confirming previously described phenotypic similarities. Lagrangian particle dispersion model simulations using finely resolved meteorological data showed that long distance dispersal events between southern Africa and Australia are indeed possible, albeit rare. Simulated urediniospore transmission events were most frequent from central South Africa (viable spore transmission on approximately 7% of all simulated release days) compared with other potential source regions in southern Africa. The study acts as a warning of possible future P. graminis f. sp. tritici dispersal events from southern Africa to Australia, which could include members of the Ug99 race group, emphasizing the need for continued surveillance on both continents.

Obesity, Body Mass Index, and Homicide.

The body mass indexes (BMIs) of 100 randomly selected homicide cases from the files of Forensic Science SA were compared to the Australian and South Australian populations. There were 70 males and 30 females (M:F = 2.3:1; age range 18-84 years; mean 42.3 years). There was a substantially lower proportion of obese individuals in the homicide population compared to the general Australian and South Australian populations (19% [vs.] 27.9% and 30%, respectively). A second group of 144 randomly selected autopsy cases where the BMI was ≥40 kg/m2 was analyzed. There were 77 males and 67 females (M:F = 1.2:1; age range 23-78 years; mean 46.7 years). The majority of deaths were natural (N = 108), with no homicides. A negative association between obesity and homicide has, therefore, been demonstrated. Reasons for the lower numbers of obese/morbidly obese individuals among homicide victims are unclear, but may include physical protection afforded by fat padding from sharp force injuries, and relative sociodemographic isolation.

Polymorphisms within the COL5A1 gene and regulators of the extracellular matrix modify the risk of Achilles tendon pathology in a British case-control study

ABSTRACTSeveral genetic loci have been associated with risk of Achilles tendon pathology (ATP) within South African and Australian populations. The aim of this study was, therefore, to evaluate eight previously implicated genetic variants in an independent British population. A total of 130 asymptomatic controls (CON) and 112 participants clinically diagnosed with ATP comprising 87 individuals with chronic Achilles tendinopathy (TEN) and 25 with Achilles tendon ruptures (RUP) were included. All participants were genotyped for variants within the COL5A1, MIR608, IL-1β, IL-6 and CASP8 genes. Primary findings implicated COL5A1 and CASP8. Three inferred allele combinations constructed from COL5A1 rs12722, rs3196378 and rs71746744 were identified as risk modifiers. The T–C–D combination was associated with increased risk of ATP (P = 0.023) and RUP (P < 0.001), the C–A–I combination was associated with increased risk of ATP (P = 0.011), TEN (P = 0.011) and RUP (P = 0.011) and the C–C–D combination was associated with decreased risk of ATP (P = 0.011) and RUP (P = 0.004). The CASP8 rs3834129 DD genotype was associated with decreased risk of TEN (P = 0.020, odds ratio: 0.45, 95% confidence interval: 0.22–0.90) and the CASP8 I–G (rs3834129–rs1045485) inferred allele combination was associated with increased risk of TEN (P = 0.031). This study further highlights the importance of polymorphisms within COL5A1 and CASP8 in the aetiology of ATP.

Defining conservation units in a complex of morphologically similar, sexually deceptive, highly endangered orchids

In plants, rapid speciation attributed to pollination strategy is unlikely to lead to obvious morphological traits for identification or large genetic differences among populations. The species-rich Australian orchid genus Caladenia contains numerous taxonomically difficult species complexes, including the reticulata complex comprising 37 species, many of which are highly endangered with narrow-range geographic distributions as a consequence of severe habitat loss. Here, we test morphological species delimitation in the reticulata complex with a multigene phylogenetic analysis, detailed population genetic analysis and comprehensive data on specific sexually deceived pollinators of species within the complex. Phylogenetic analysis revealed limited species-level resolution within the reticulata complex and no difference between species with different pollinators. Microsatellite data confirmed this result for Victorian populations of the reticulata complex but revealed significant isolation by distance between Victorian and South Australian populations. Pollinator preference tests indicated that 10 reticulata complex morphospecies all shared the thynnid wasp pollinator Phymatothynnus nitidus, yet the microsatellite analysis revealed only limited differentiation between those and three species within the reticulata complex shown to employ different pollinators. The combined molecular and ecological approach revealed that morphospecies that share the same pollinator are likely to be the same biological species. Conservation and taxonomic focus should shift from a ‘microspecies’ approach to capturing key populations across the geographic range of the Caladenia reticulata biological species which will greatly assist the allocation of limited conservation resources.

Origin, migration routes and worldwide population genetic structure of the wheat yellow rust pathogen Puccinia striiformis f.sp. tritici.

Analyses of large-scale population structure of pathogens enable the identification of migration patterns, diversity reservoirs or longevity of populations, the understanding of current evolutionary trajectories and the anticipation of future ones. This is particularly important for long-distance migrating fungal pathogens such as Puccinia striiformis f.sp. tritici (PST), capable of rapid spread to new regions and crop varieties. Although a range of recent PST invasions at continental scales are well documented, the worldwide population structure and the center of origin of the pathogen were still unknown. In this study, we used multilocus microsatellite genotyping to infer worldwide population structure of PST and the origin of new invasions based on 409 isolates representative of distribution of the fungus on six continents. Bayesian and multivariate clustering methods partitioned the set of multilocus genotypes into six distinct genetic groups associated with their geographical origin. Analyses of linkage disequilibrium and genotypic diversity indicated a strong regional heterogeneity in levels of recombination, with clear signatures of recombination in the Himalayan (Nepal and Pakistan) and near-Himalayan regions (China) and a predominant clonal population structure in other regions. The higher genotypic diversity, recombinant population structure and high sexual reproduction ability in the Himalayan and neighboring regions suggests this area as the putative center of origin of PST. We used clustering methods and approximate Bayesian computation (ABC) to compare different competing scenarios describing ancestral relationship among ancestral populations and more recently founded populations. Our analyses confirmed the Middle East-East Africa as the most likely source of newly spreading, high-temperature-adapted strains; Europe as the source of South American, North American and Australian populations; and Mediterranean-Central Asian populations as the origin of South African populations. Although most geographic populations are not markedly affected by recent dispersal events, this study emphasizes the influence of human activities on recent long-distance spread of the pathogen.

Recovery of South Australian rabbit populations from the impact of rabbit haemorrhagic disease

Context Recovery of Australian rabbit populations from the impact of rabbit haemorrhagic disease virus (RHDV) contrasts with more prolonged suppression of wild rabbits in Europe, and has been widely discussed in the scientific community, but not yet documented in formal scientific literature. The underlying causes of recovery remain unclear, but resistance to RHDV infection has been reported in laboratory studies of wild-caught rabbits. Aims We document numerical changes in two South Australian wild rabbit populations that were initially suppressed by RHDV, and examine serological data to evaluate several alternative hypotheses for the cause of recovery. Methods Rabbit numbers were assessed from spotlight transect counts and dung mass transects between 1991 and 2011, and age and RHDV antibody sero-prevalence were estimated from rabbits shot in late summer. Key results Rabbit numbers were heavily suppressed by RHDV between 1995 and 2002, then increased 5- to 10-fold between 2003 and 2010. During the period of increase, annual RHDV infection rates remained stable or increased slightly, average age of rabbits remained stable and annual rainfall was below average. Conclusions Rabbit populations recovered but neither avoidance of RHDV infection, gradual accumulation of long-lived RHD-immune rabbits, nor high pasture productivity were contributing factors. This leaves increased annual survival from RHDV infection as the most likely cause of recovery. Implications Previously documented evidence of resistance to RHDV infection may be of little consequence to post-RHD recovery in rabbit numbers, unless the factors that influence the probability of infection also shape the course of infection and affect survival of infected rabbits.

Age-dependent changes in gross and histological morphology of the thyroid gland in South Australian koalas (Phascolarctos cinereus)

Studies characterising the thyroid gland structure of koalas are limited and have not previously been undertaken in South Australian populations. Hence, this study aimed to describe the thyroid gland morphology of koalas from the Mount Lofty region, South Australia. Results showed that thyroid gland morphology was highly variable between individual koalas (n = 36), ranging from that considered typical for healthy mammals, in which small colloid-filled follicles were lined by cuboidal epithelium, to that consistent with colloidal goitre, in which macrofollicles distended with colloid were lined by flattened epithelium. Juvenile koalas more frequently showed typical thyroid gland morphology than adults, with significantly higher thyroid follicle density (P &lt; 0.05) and a higher proportion of follicles lined by cuboidal epithelium compared with flattened epithelium (P &lt; 0.05). Thyroid glands of most adult koalas were characterised by colloidal macrofollicles (P &lt; 0.01), and classified as colloidal goitre. There were no significant differences in thyroid gland morphology based on health status or sex of koalas. These findings suggest that an age-dependent colloidal goitre occurs in adult koalas, which is unlikely to have ill effects and may be associated with the low metabolic rate of this species, or exposure to a dietary goitrogen.

Variants within the COMP and THBS2 genes are not associated with Achilles tendinopathy in a case-control study of South African and Australian populations

Cartilage oligomeric matrix protein is a structural protein of the extracellular matrix, while thrombospondin-2 is a matricellular protein involved in cell–matrix interactions. Recent studies have shown that genetic variation is a significant risk factor for Achilles tendinopathy, and the genes encoding cartilage oligomeric matrix protein (COMP) and thrombospondin-2 (THBS2) were identified as good candidate genes for association with Achilles tendinopathy. This study aimed to test the association of sequence variants within these candidate genes with the risk of Achilles tendinopathy in participants from South Africa (SA) and Australia (AUS). Three-hundred and forty (133 SA; 207 AUS) control participants with no history of Achilles tendinopathy and 178 (94 SA; 84 AUS) participants clinically diagnosed with Achilles tendinopathy were genotyped for five single nucleotide polymorphisms within the COMP and THBS2 genes in this case-control study. There was no difference in genotype distributions between control and tendinopathy groups for either the THBS2 variants rs9505888, rs6422747 and rs9283850, or the COMP variants rs730079 and rs28494505 in the SA and AUS populations. As the selection of COMP and THBS2 as candidate genes was hypothesis driven, based on biological function, the possibility that other variants within these genes are associated with Achilles tendinopathy cannot be excluded.

Population Divergence in Venom Bioactivities of Elapid Snake Pseudonaja textilis: Role of Procoagulant Proteins in Rapid Rodent Prey Incapacitation

This study looked at how toxic proteins in venoms of adult Australian eastern Brown snakes Pseudonaja textilis from South Australian and Queensland populations interact with physiological functions of the lab SD rat Rattus norvegicus. Circulatory collapse and incoagulable blood occurred instantly after injection of venom under the dorsal skin of anaesthetised and mechanically ventilated rats in an imitation of a P. textilis bite. Intravenous injection of purified P. textilis (Mackay, QLD) venom prothrombin activator proteins caused instant failure of circulation, testifying of high toxicity of these proteins and suggesting their role in rapid incapacitation of rodent prey. The hypothesis is further supported by circulatory collapse occurring instantly despite artificial respiration in envenomed rats and the finding of extremely high venom procoagulant potency in rat plasma. LC-MS and physiology assays revealed divergent venom composition and biological activity of South Australian (Barossa locality) and Queensland (Mackay locality) populations, which may be driven by selection for different prey. The Queensland venom of P. textilis was found to be more procoagulant and to exhibit predominately presynaptic neurotoxicity, while the South Australian venom contained diverse postsynaptic type II and III α-neurotoxins in addition to the presynaptic neurotoxins and caused significantly faster onset of neuromuscular blockade in the rat phrenic nerve-diaphragm preparation. LC-MS analysis found evidence of multiple coagulation factor X-like proteins in P. textilis venoms, including a match to P. textilis coagulation factor X isoform 2, previously known to be expressed only in the liver.

Elucidating the native sources of an invasive tree species, Acacia pycnantha, reveals unexpected native range diversity and structure.

Understanding the introduction history of invasive plant species is important for their management and identifying effective host-specific biological control agents. However, uncertain taxonomy, intra- and interspecific hybridization, and cryptic speciation may obscure introduction histories, making it difficult to identify native regions to explore for host-specific agents. The overall aim of this study was to identify the native source populations of Acacia pycnantha, a tree native to south-eastern Australia and invasive in South Africa, Western Australia and Portugal. Using a phylogeographical approach also allowed an exploration of the historical processes that have shaped the genetic structure of A. pycnantha in its native range. Nuclear (nDNA) and plastid DNA sequence data were used in network and tree-building analyses to reconstruct phylogeographical relationships between native and invasive A. pycnantha populations. In addition, mismatch distributions, relative rates and Bayesian analyses were used to infer recent demographic processes and timing of events in Australia that led to population structure and diversification. The plastid network indicated that Australian populations of A. pycnantha are geographically structured into two informally recognized lineages, the wetland and dryland forms, whereas the nuclear phylogeny showed little geographical structure between these two forms. Moreover, the dryland form of A. pycnantha showed close genetic similarity to the wetland form based on nDNA sequence data. Hybrid zones may explain these findings, supported here by incongruent phylogenetic placement of some of these taxa between nuclear and plastid genealogies. It is hypothesized that habitat fragmentation due to cycles of aridity inter-dispersed with periods of abundant rainfall during the Pleistocene (approx. 100 kya) probably gave rise to native dryland and wetland forms of A. pycnantha. Although the different lineages were confined to different ecological regions, we also found evidence for intraspecific hybridization in Victoria. The invasive populations in Portugal and South Africa represent wetland forms, whereas some South African populations resemble the Victorian dryland form. The success of the biological control programme for A. pycnantha in South Africa may therefore be attributed to the fact that the gall-forming wasp Trichilogaster signiventris was sourced from South Australian populations, which closely match most of the invasive populations in South Africa.

“Cycliplex PCR” confirmation of Fusobacterium necrophorum isolates from captive wallabies: A rapid and accurate approach

Isolation and identification of obligate anaerobic bacteria is labour intensive and time consuming. This has led to the increased application of molecular tools to circumvent part of this problem. We report here the development of a rapid, accurate and cost-effective method to isolate and identify Fusobacterium necrophorum species from South Australian wallaby populations using a supplemented medium (BHIRS) in conjunction with a “Cycliplex PCR” method which involves a stepwise-selective amplification of target PCR products. This report demonstrates the complementation of phenotypic characterization by PCR for accurate and fast identification of F. necrophorum isolates from wildlife origin.