Social networks and the spread of Salmonella in a sleepy lizard population.

Although theoretical models consider social networks as pathways for disease transmission, strong empirical support, particularly for indirectly transmitted parasites, is lacking for many wildlife populations. We found multiple genetic strains of the enteric bacterium Salmonella enterica within a population of Australian sleepy lizards (Tiliqua rugosa), and we found that pairs of lizards that shared bacterial genotypes were more strongly connected in the social network than were pairs of lizards that did not. In contrast, there was no significant association between spatial proximity of lizard pairs and shared bacterial genotypes. These results provide strong correlative evidence that these bacteria are transmitted from host to host around the social network, rather than that adjacent lizards are picking up the same bacterial genotype from some common source.

Managed relocation as an adaptation strategy for mitigating climate change threats to the persistence of an endangered lizard.

The distributional ranges of many species are contracting with habitat conversion and climate change. For vertebrates, informed strategies for translocations are an essential option for decisions about their conservation management. The pygmy bluetongue lizard, Tiliqua adelaidensis, is an endangered reptile with a highly restricted distribution, known from only a small number of natural grassland fragments in South Australia. Land-use changes over the last century have converted perennial native grasslands into croplands, pastures and urban areas, causing substantial contraction of the species' range due to loss of essential habitat. Indeed, the species was thought to be extinct until its rediscovery in 1992. We develop coupled-models that link habitat suitability with stochastic demographic processes to estimate extinction risk and to explore the efficacy of potential climate adaptation options. These coupled-models offer improvements over simple bioclimatic envelope models for estimating the impacts of climate change on persistence probability. Applying this coupled-model approach to T. adelaidensis, we show that: (i) climate-driven changes will adversely impact the expected minimum abundance of populations and could cause extinction without management intervention, (ii) adding artificial burrows might enhance local population density, however, without targeted translocations this measure has a limited effect on extinction risk, (iii) managed relocations are critical for safeguarding lizard population persistence, as a sole or joint action and (iv) where to source and where to relocate animals in a program of translocations depends on the velocity, extent and nonlinearities in rates of climate-induced habitat change. These results underscore the need to consider managed relocations as part of any multifaceted plan to compensate the effects of habitat loss or shifting environmental conditions on species with low dispersal capacity. More broadly, we provide the first step towards a more comprehensive framework for integrating extinction risk, managed relocations and climate change information into range-wide conservation management.

Evaluation of offspring size-number invariants in 12 species of lizard.

The optimal division of resources into offspring size vs. number is one of the classic problems in life-history evolution. Importantly, models that take into account the discrete nature of resource division at low clutch sizes suggest that the variance in offspring size should decline with increasing clutch size according to an invariant relationship. We tested this prediction in 12 species of lizard with small clutch sizes. Contrary to expectations, not all species showed a negative relationship between variance in offspring size and clutch size, and the pattern significantly deviated from quantitative predictions in five of the 12 species. We suggest that the main limitation of current size-number models for small clutch sizes is that they rely on assumptions of hierarchical allocation strategies with independence between allocation decisions. Indeed, selection may favour alternative mechanisms of reproductive allocation that avoid suboptimal allocation imposed by the indivisible fraction at low clutch sizes.

Unique biological rhythm in the reproductive behaviour of female ticks of reptiles.

We report the discovery of a biological rhythm in the reproductive behaviour of the tick Bothriocroton hydrosauri that was absent in Amblyomma limbatum, a species that occurs on the same species of reptile host. Female B. hydrosauri mated in autumn or winter delayed oviposition until the following spring, while there was no diapause in conspecific females mated in spring or early summer. Initiation of ovipositional diapause in ticks is usually related to photoperiodic stimuli, but this was not the case for B. hydrosauri. The sinusoidal pattern in pre-oviposition times of B. hydrosauri females mated in different months in the laboratory suggests an internal seasonal time-keeping mechanism. We hypothesize that hormones imbibed by females during their bloodmeal may provide environmental cues associated with the induction of diapause. Irrespective of the mechanism underlying the rhythm, diapause by B. hydrosauri females mated during autumn or winter is of adaptive advantage because it synchronizes oviposition with favourable environmental conditions for egg hatching and increases the chance of larvae finding a host. The lack of a similar biological rhythm in A. limbatum may be a reflection of the different environmental conditions this species experiences throughout most of its range as compared with B. hydrosauri.

Seasonal and spatial dynamics of ectoparasite infestation of a threatened reptile, the tuatara (Sphenodon punctatus).

The conservation of threatened vertebrate species and their threatened parasites requires an understanding of the factors influencing their distribution and dynamics. This is particularly important for species maintained in conservation reserves at high densities, where increased contact among hosts could lead to increased rates of parasitism. The tuatara (Sphenodon punctatus) (Reptilia: Sphenodontia) is a threatened reptile that persists at high densities in forests (approximately 2700 tuatara/ha) and lower densities in pastures and shrubland (< 200 tuatara/ha) on Stephens Island, New Zealand. We investigated the lifecycles and seasonal dynamics of infestation of two ectoparasites (the tuatara tick, Amblyomma sphenodonti, and trombiculid mites, Neotrombicula sp.) in a mark-recapture study in three forest study plots from November 2004 to March 2007, and compared infestation levels among habitat types in March 2006. Tick loads were lowest over summer and peaked from late autumn (May) until early spring (September). Mating and engorgement of female ticks was highest over spring, and larval tick loads subsequently increased in early autumn (March). Nymphal tick loads increased in September, and adult tick loads increased in May. Our findings suggest the tuatara tick has a 2- or 3-year lifecycle. Mite loads were highest over summer and autumn, and peaked in March. Prevalences (proportion of hosts infected) and densities (estimated number of parasites per hectare) of ticks were similar among habitats, but tick loads (parasites per host) were higher in pastures than in forests and shrub. The prevalence and density of mites was higher in forests than in pasture or shrub, but mite loads were similar among habitats. We suggest that a higher density of tuatara in forests may reduce the ectoparasite loads of individuals through a dilution effect. Understanding host-parasite dynamics will help in the conservation management of both the host and its parasites.

A comparison of the reproductive parameters of females of two reptile tick species.

In comparisons of females of two reptile tick species Aponomma hydrosauri and Amblyomma limbatum, Ap. hydrosauri was initially larger, and after mating on the host engorged faster and remained attached for a shorter time before completing engorgement and detaching. Amb. limbatum had a longer period of engorgement, and achieved a greater engorged weight. Engorged Amb. limbatum females laid significantly more eggs than equivalent sized Ap. hydrosauri. Although the two species are ecologically similar and were collected from the same site for this study, their reproductive differences probably reflect adaptations to different conditions in their largely allopatric ranges.

Influence of environmental factors on oviposition and egg development in Amblyomma limbatum and Aponomma hydrosauri (Acari: Ixodidae).

This study examined the influence of temperature and light on the length of the pre-oviposition period of engorged females of two Australian ixodid ticks, Amblyomma limbatum and Aponomma hydrosauri. The hatching success and development time of eggs of both species were also compared at different temperatures and relative humidities. Darkness was found to have no effect on the duration of the pre-oviposition time or reproductive output of females of either species. In contrast, the preoviposition period of females of both species decreased with increasing temperature. Amb. limbatum females had shorter pre-oviposition periods than Ap. hydrosauri at all temperatures examined. Temperature and relative humidity had a marked effect on the hatching success of eggs. Eggs of both species had reduced hatching success at low relative humidities. Eggs failed to hatch at temperatures below 21 degrees C. Ap. hydrosauri eggs also failed to hatch at 34 degrees C while Amb. limbatum eggs failed to hatch at 36 degrees C. Within the range of temperatures suitable for egg development, the hatching times of eggs of both species decreased with increasing temperature. Amb. limbatum eggs developed faster than Ap. hydrosauri eggs at temperatures greater than 25 degrees C, but slower at cooler temperatures. These differences in the duration of their preoviposition period, and the responses of females and their eggs to different temperatures and relative humidities correlate with the different climates the two species experience throughout most of their distributional range.

Interspecific differences in the movements of female ticks on reptiles.

Movements of females of two reptile ticks, Amblyomma limbatum and Aponomma hydrosauri were studied to determine whether the behaviour of the species on hosts was similar or different. Experiments were conducted to determine if the movement of females on hosts prior to mating was influenced by the delayed attachment of conspecific males. A field experiment was also undertaken to determine whether data obtained during laboratory experiments reflected the behaviour of females on hosts in the field. Both in laboratory and field experiments, a significantly greater proportion of Ap. hydrosauri females changed their site of attachment prior to mating. Individual Ap. hydrosauri females made significantly more positional changes than Amb. limbatum females. These interspecific differences occurred irrespective of the absence of conspecific males or presence of non-conspecific adults on hosts. These data may have important ecological implications in relation to the success of colonizing females at their parapatric boundary.

Influence of temperature and relative humidity on the moulting success of Amblyomma limbatum and Aponomma hydrosauri (Acari: Ixodidae) larvae and nymphs.

This study compared the duration of the moulting periods of engorged larvae and nymphs of the ixodid ticks, Amblyomma limbatum and Aponomma hydrosauri, at different temperature/relative humidity regimes, and examined the relationships between the engorged weight of ticks and their weights after moulting. The results showed that for each species, there was a significant relationship between the weights of unfed nymphs and engorged larvae, and the weights of unfed adults and engorged nymphs. The weight of engorged nymphs was also a good indicator of their sex, with female ticks having heavier weights as engorged nymphs. Temperature and relative humidity had a marked effect on the moulting success of engorged ticks of both species. Aponomma hydrosauri larvae and nymphs were able to moult at lower temperatures than Amb. limbatum but most ticks, except Ap. hydrosauri larvae, failed to moult at 13 degrees C. Additionally, there was a marked decrease in the pre-moult times of ticks at higher temperatures, with larvae taking less time to moult than nymphs. At temperatures greater than 21 degrees C, Amb. limbatum took less time to moult than Ap. hydrosauri but this interspecific difference was less marked for nymphs. The interspecific differences in the responses of engorged larvae and nymphs to different temperatures and relative humidities correlated with interspecific differences in off-host behaviour and with the different climates the two species experience throughout most of their distributional range.

The impact of tick parasites on the behaviour of the lizard Tiliqua rugosa.

Populations of the Australian sleepy lizard, Tiliqua rugosa, near Mt. Mary, South Australia carry natural infestations of two tick species Aponomma hydrosauri and Amblyomma limbatum. In field experiments at two sites, 18 km apart, lizards with experimentally increased tick loads had smaller home ranges, moved shorter distances in a day, and were found basking more but moving less often than lizards from which ticks were experimentally removed. The results were consistent for adult lizards in two years, and for sub-adults in a third year. Laboratory trials showed that juvenile lizards that had tick infestations had lower sprint speeds than uninfested siblings, and that adults with tick infestations had less endurance than those that were uninfested. The results contrast with those of a previous survey that showed that lizards with high tick loads had greater body size and remained longer at a site, but indicate that there may be a balance, for lizards, between the fitness advantages in occupying habitats with high-quality resources, and the costs from parasites that also prefer those habitats.

Water movements, tadpole competition and limits to the distribution of the frogs Ranidella riparia and R. signifera.

Ranidella signifera and R. riparia are leptodactylid frogs with largely allopatric distributions which contact and narrowly overlap in the Flinders Ranges of South Australia. One hypothesisto explain the distributional limits is that each species cannot persist in the creeks beyond its range. Survival and growth of tadpoles were measured in enclosures in three creeks, one on either side of the overlap zone and one in it. When each species was kept alone there were no differences between them either in the slow water typical of R. signifera creeks or in fast moving water typical of R. riparia creeks. When the two species were kept together there were no consistent differences in survival or growth in any creek where water flow was shielded in the experimental enclosures. Where water flow was not shielded through the enclosures R. riparia tadpoles had significantly higher survival than R. signifera. A form of interaction is proposed where R. signifera is more frequently exposed to the water current when R. riparia is present. This may explain why R. signifera does not extend its distribution into the adjacent fast rocky creeks occupied by R. riparia, but the experiments did not explain the limitation of the range of R. riparia.

No competition for resources between two tick species at their parapatric boundary.

Aponomma hydrosauri and Amblyomma limbatum are tick species with the same major host species, the sleepy lizard Trachydosaurus rugosus, but parapatric distributions in South Australia. Near Mt. Mary one species is abruptly replaced by the other over one kilometre. There is substantial overlap in feeding sites on the hosts. This suggested that interspecific competition, during feeding, may be an important mechanism preventing further overlap of the distributions of the two species. However, in the overlap region there was no resource shortage, and each species infested hosts independently of the other species. Also in laboratory experiments at the highest densities encountered in the field, larvae and nymphs of each species attached and engorged equally well whether alone or mixed with the other species. These data refuted the, hypothesis of interspecific competition between the two species while feeding.

Niche changes between parasite populations: An example from ticks on reptiles.

Two Australian tick species Aponomma hydrosauri and Amblyomma albolimbatum have the same major host species, the lizard Trachydosaurus rugosus. While females of Amb. albolimbatum are most often attached in the ears and on the neck of their hosts, Ap. hydrosauri females prefer to attach further back, under the forearms and on the back. Males show the same interspecific difference but there is also a difference between populations. Ap. hydrosauri males from populations in contact with Amb. albolimbatum attach more often in posterior positions than Ap. hydrosauri males from populations isolated from Amb. albolimbatum. These differences were found in both field populations and laboratory reared ticks. Phylogenetic evidence suggests that the change in male attachment site between Ap. hydrosauri populations followed the colonization of T. rugosus. We propose that the most likely reason for the change of attachment sites has been interspecific interactions with Amb. albolimbatum and that competition has been for space for efficient reception of female signals.

Habitat selection in tadpoles of Ranidella signifera and R. riparia (Anura: Leptodactylidae).

Two leptodactylid frog species Ranidella signifera and R. riparia occupy adjacent but different habitat types in the Flinders Ranges of South Australia. R. signifera is found in slow-flowing creeks with mud or sand substrates. R. riparia occupies fast-flowing rocky creeks. The two coexist in a narrow overlap zone with heterogeneous habitats. Laboratory experiments were used to test habitat preferences of tadpoles in a tub with sand and rock substrates. R. riparia was most commonly found on the rock substrate. R. signifera showed no distinct preference for sand, but when mixed with R. riparia increased its use of the habitat in the spaces between and under rocks. This was particularly apparent in flowing water where 81% of all R. signifera tadpoles were observed in this sheltered habitat.The data suggest that R. signifera tadpoles have reduced fitness in flowing water because they cannot exploit exposed feeding sites where most of the algae grow, and that interspecific interactions with R. riparia tadpoles reduce their fitness further. This helps to explain why R. signifera do not extend their distribution into R. riparia occupied creeks. The data do not explain why R. riparia do not colonize the slow muddy creeks occupied by R. signifera.

Reproductive interference between three parapatric species of reptile tick.

Reproductive interference between three species of reptile tick, Aponomma hydrosauri, Amblyomma albolimbatum and Amb. limbatum was investigated. Adults of two species attach together on the same lizard hosts in narrow overlap zones at parapatric boundaries between species, providing opportunities for interference. The possibility of reproductive interference was suggested because of similarities in the sexual communication systems.Three forms of interference were found in laboratory experiments. Firstly male movement to search for sexually receptive females was inhibited when females of two species were present on the same host, probably because of signal interference. Secondly, a nonspecific, short range attractant pheromone led males into non-conspecific courtship attempts which reduced the time for conspecific courtships. Thirdly Amb. albolimbatum males physically blocked the genitalia of female Ap. hydrosauri after unsuccessful non-conspecific courtship attempts. Field data showed this behaviour was common in both Amblyomma species after conspecific matings. Reproductive interference would reduce the fitness of each species in sympatry, and may contribute to the maintenance of the narrow parapatric boundaries.

Host detection by four Australian tick species.

The behavior of 4 tick species in the presence of their dominant host species was examined. Nymphs and adult ticks could locate a host from greater distances than the larvae. Adult Aponomma concolor were able to locate their mammalian host (echidna) from distances greater than 3 reptile-infesting species could locate their hosts. The results suggested that all 4 tick species detect hosts passively, which may be related to the relative scarcity of hosts and their irregular use of refuge sites.

Clustering of related individuals in a population of the Australian lizard, Egernia frerei.

Stable social aggregations are rarely recorded in lizards, but have now been reported from several species in the Australian scincid genus Egernia. Most of those examples come from species using rock crevice refuges that are relatively easy to observe. But for many other Egernia species that occupy different habitats and are more secretive, it is hard to gather the observational data needed to deduce their social structure. Therefore, we used genotypes at six polymorphic microsatellite DNA loci of 229 individuals of Egernia frerei, trapped in 22 sampling sites over 3500 ha of eucalypt forest on Fraser Island, Australia. Each sampling site contained 15 trap locations in a 100 x 50 m grid. We estimated relatedness among pairs of individuals and found that relatedness was higher within than between sites. Relatedness of females within sites was higher than relatedness of males, and was higher than relatedness between males and females. Within sites we found that juvenile lizards were highly related to other juveniles and to adults trapped at the same location, or at adjacent locations, but relatedness decreased with increasing trap separation. We interpreted the results as suggesting high natal philopatry among juvenile lizards and adult females. This result is consistent with stable family group structure previously reported in rock dwelling Egernia species, and suggests that social behaviour in this genus is not habitat driven.

Optimized blood cell profiling method for genomic biomarker discovery using high-density microarray.

High-quality biomarkers for disease progression, drug efficacy and toxicity liability are essential for improving the efficiency of drug discovery and development. The identification of drug-activity biomarkers is often limited by access to and the quantity of target tissue. Peripheral blood has increasingly become an attractive alternative to tissue samples from organs as source for biomarker discovery, especially during early clinical studies. However, given the heterogeneous blood cell population, possible artifacts from ex vivo activations, and technical difficulties associated with overall performance of the assay, it is challenging to profile peripheral blood cells directly for biomarker discovery. In the present study, Applied BioSystems' blood collection system was evaluated for its ability to isolate RNA suitable for use on the Affymetrix microarray platform. Blood was collected in a TEMPUS tube and RNA extracted using an ABI-6100 semi-automated workstation. Using human and rat whole blood samples, it was demonstrated that the RNA isolated using this approach was stable, of high quality and was suitable for Affymetrix microarray applications. The microarray data were statistically analysed and compared with other blood protocols. Minimal haemoglobin interference with RNA labelling efficiency and chip hybridization was found using the TEMPUS tube and extraction method. The RNA quality, stability and ease of handling requirement make the TEMPUS tube protocol an attractive approach for expression profiling of whole blood to support target and biomarker discovery.

Reproductive interactions between two Australian reptile tick species.

In South Australia the two tick species Amblyomma limbatum and Aponomma hydrosauri share the same common reptile host species, but have allopatric distributions which abut along a narrow parapatric boundary. Reproductive interference is a mechanism that has previously been suggested could contribute to maintaining the boundary. Populations of each species were established in pens within the range of Aponomma hydrosauri. Pens held either each species alone, or the two species together. The performance of females in those pens was monitored over 28 months. There was no indication that the proportion of attached females which mated and engorged was reduced by the presence of heterospecifics. There was no indication that the time taken to mate, engorge and detach was any longer in the presence of heterospecifics. The experiment did not support the hypothesis that reproductive interference contributes to maintaining the parapatric boundary. However, Amblyomma limbatum in the pens had a shorter season of reproductive activity, and achieved much smaller numbers of reproductive females. This may inhibit successful colonization of cooler habitats to the south of its distribution.

Prevalence of infection by the protozoan Hemolivia mariae in ticks.

This paper considers the prevalence of natural infections of the protozoan Hemolivia mariae, in its hosts the Australian sleepy lizard, Tiliqua rugosa, and the ixodid tick species, Amblyomma limbatum. We investigate whether the protozoan may be influencing the tick population in the field, by comparing the observed prevalence of infection in ticks with the prevalence expected from known transmission dynamics. The prevalence of infection in nymphs was similar to the expected prevalence, but the observed prevalence in adults was higher than expected. These results provide no evidence for infection-induced mortality in ticks. We also found that tick loads on infected and uninfected lizards were not significantly different and, overall, infected lizards were as likely to be tick-infested as uninfected lizards. However, infected lizards were less likely to be found carrying female ticks. On balance, the evidence did not strongly support the hypothesis that ticks avoid feeding on infected lizards. We use known parameters of H. mariae transmission to estimate the rate of tick ingestion that may be required to sustain the observed prevalences in the field.