Molecular cloning of natriuretic peptides from the heart of reptiles : loss of ANP in diapsid reptiles and birds

Atrial natriuretic peptide (ANP) and B-type NP (BNP) are hormones involved in homeostatic control of body fluid and cardiovascular regulation. Both ANP and BNP have been cloned from the heart of mammals, amphibians, and teleost fishes, while an additional cardiac peptide, ventricular NP, has been found in selected species of teleost fish. However, in chicken, BNP is the primary cardiac peptide identified thus far. In contrast, the types of NP/s present in the reptilian heart are unknown, representing a considerable gap in our understanding of NP evolution. In the present study, we cloned and sequenced a BNP cDNA from the atria of representative species of reptile, including crocodile, lizard, snake, and tortoise. In addition, we cloned BNP from the pigeon atria. The reptilian and pigeon BNP cDNAs had ATTTA repeats in the 3′ untranslated region, as observed in all vertebrate BNP mRNAs. A high sequence homology was evident when comparing reptile and pigeon preproBNP with the previously identified chicken preproBNP. In particular, the predicted mature BNP-29 was identical between crocodile, tortoise, and chicken, with pigeon having a single amino acid substitution; lizard and snake BNP had seven and nine substitutions, respectively. Furthermore, an ANP cDNA could only be cloned from the tortoise atria. Since ANP was not isolated from the heart of any non-chelonian reptile and appears to be absent in birds, we propose that the ANP gene has been lost after branching of the turtles in the amniote line. This data provides new avenues for research on NP function in reptiles.

Molecular biology of natriuretic peptides in reptiles and birds

The heart produces natriuretic peptides that are critical regulators of blood pressure and renal function. This study examined the molecular evolution of natriuretic peptides in vertebrates and discovered novel forms of the peptides in birds. The research outcomes advanced our knowledge of the importance of these peptides in animal physiology.

Fire, energy, and the ecology of reptiles in semi-arid Australia

‬Fire dependent ecosystems cover over half of the world's land surface. Understanding the factors that determine the distribution of fauna in these systems is essential to biodiversity conservation. This thesis explores the ecology of reptiles in a fire-prone region.

Life in the really slow lane : loggerhead sea turtles mature late relative to other reptiles

1. Age at maturity is hard to estimate for species that cannot be directly marked or observed throughout their lives and yet is a key demographic parameter that is needed to assess the conservation status of endangered species. 2. For loggerhead turtles (Caretta caretta) in the North Atlantic and North Pacific, juvenile growth rates (c. 10 cm year−1) were calculated by examining size increases during transoceanic journeys; durations of which were estimated from satellite-tracked Lagrangian surface drifter buoy trajectories. 3. Lagrangian-derived growth estimates were used in a weighted loglinear model of size-specific growth rates for loggerhead turtles and combined with newly available information on size at maturity to estimate an age at maturity of 45 years (older than past estimates). 4. By examining the age at maturity for 79 reptile species, we show that loggerhead turtles, along with other large-bodied Testudine (turtle and tortoise) species, take longer to reach maturity than other reptile species of comparable sizes. This finding heightens concern over the future sustainability of turtle populations. By maturing at an old age, sea turtles will be less resilient to anthropogenic mortality than previously suspected.

Molecular and morphological description of a Hepatozoon species in reptiles and their ticks in the Northern Territory, Australia

Ticks, representing 3 species of Amblyomma, were collected from the water python (Liasis fuscus) and 3 additional reptile species in the Northern Territory, Australia, and tested for the presence of Hepatozoon sp., the most common blood parasites of snakes. In addition, blood smears were collected from 5 reptiles, including the water python, and examined for the presence of the parasite. Hepatozoon sp. DNA was detected in all tick and reptile species, with 57.7% of tick samples (n = 187) and 35.6% of blood smears (n=35) showing evidence of infection. Phylogenetic analysis of the 18S rRNA gene demonstrated that half of the sequences obtained from positive tick samples matched closest with a Hepatozoon species previously identified in the water python population. The remaining sequences were found to be more closely related to mammalian and amphibian Hepatozoon species. This study confirms that species of Amblyomma harbor DNA of the same Hepatozoon species detected in the water pythons. The detection of an additional genotype suggests the ticks may be exposed to 2 Hepatozoon species, providing further opportunity to study multiple host-vector-parasite relationships

Reptiles in restored agricultural landscapes: the value of linear strips, patches and habitat condition

Habitat restoration, including revegetation of linear strips and enlargement of remnant patches, may benefit native fauna in highly fragmented landscapes. Such restoration has occurred around the world, even though the relative importance of strips and patches of vegetation remains controversial. Using reptile communities from south-eastern Australia, we assessed the conservation value of revegetation in strips and alongside remnant patches compared with remnant vegetation and cleared roadsides. We also examined the distance that reptiles occurred from remnant patches into linear vegetation. We found that reptile species richness and counts did not substantially differ between revegetated, remnant and cleared habitats, or between linear strip and patch treatments. This may indicate that species sensitive to land clearing have already been lost from the landscape. These results imply that if specialist species have already been lost, we may be unable to measure the effects of agriculture on biodiversity. Furthermore, revegetation with the expectation that fauna will recolonize may be unrealistic and translocations may be necessary. Unexpectedly, we recorded higher species richness and counts of rare reptile species in remnant linear strips as distance from remnant patches increased. Ground-layer attributes were important for increasing reptile species richness and counts and in structuring reptile communities, explaining approximately three times as much variation as remnant shape or vegetation type (remnant, revegetated, cleared). Management agencies should protect and effectively manage remnant linear strips if rarer reptiles are to be retained, paying particular attention to ground-layer attributes. The decision to include ground layers in future revegetation activities will be more important than the shape of restored areas.

Concordance in phylogeography and ecological niche modelling identify dispersal corridors for reptiles in arid Australia

Aim: Using the rock-specialist agamid Ctenophorus caudicinctus as a model, we test hypothesized biogeographical dispersal corridors for lizards in the Australian arid zone (across the western sand deserts), and assess how these dispersal routes have shaped phylogeographical structuring. Location: Arid and semi-arid Australia. Methods: We sequenced a c. 1400 bp fragment of mtDNA (ND2) for 134 individuals of C. caudicinctus as well as a subset of each of the mtDNA clades for five nuclear loci (BDNF, BACH1, GAPD, NTF3, and PRLR). We used phylogenetic methods to assess biogeographical patterns within C. caudicinctus, including relaxed molecular clock analyses to estimate divergence times. Ecological niche modelling (Maxent) was employed to estimate the current distribution of suitable climatic envelopes for each lineage. Results: Phylogenetic analyses identified two deeply divergent mtDNA clades within C. caudicinctus - an eastern and western clade - separated by the Western Australian sand deserts. However, divergences pre-date the Pleistocene sand deserts. Phylogenetic analyses of the nuclear DNA data sets generally support major mtDNA clades, suggesting past connections between the western C. c. caudicinctus populations in far eastern Pilbara (EP) and the lineages to the east of the sand deserts. Ecological niche modelling supports the continued suitability of climatic conditions between the Central Ranges and the far EP for C. c. graafi. Main conclusions: Estimates of lineage ages provide evidence of divergence between eastern and western clades during the Miocene with subsequent secondary contact during the Pliocene. Our results suggest that this secondary contact occurred via dispersal between the Central Ranges and the far EP, rather than the more southerly Giles Corridor. These events precede the origins of the western sand deserts and divergence patterns instead appear associated with Miocene and Pliocene climate change.

Nitric oxide control of lower vertebrate blood vessels by vasomotor nerves

In mammals, much is understood about the endothelial and neural NO control mechanisms in the vasculature. In contrast, NO control of blood vessels in lower vertebrates is poorly understood, with the majority of research focusing on the presence of an endothelial NO system; however, its presence remains controversial. This study examined the mechanisms by which NO regulates the large blood vessels of non-mammalian vertebrates. In all species examined, the arteries and veins contained a plexus of NOS-positive perivascular nerves that included nerve bundles and fine, varicose nerve terminals. However, in the large arteries and veins of various species of fishes and amphibians, no anatomical evidence was found for endothelial NOS using both NADPH-diaphorase and eNOS immunohistochemistry. In contrast, perinuclear NOS staining was readily apparent in blue-tongue lizard, pigeon and rat, which suggested that eNOS first appeared in reptiles. Physiological analysis of NO signalling in the vascular smooth muscle of short-finned eel and cane toad could not find any evidence for endothelial NO signalling. In contrast, it appears that activation of the nitrergic vasomotor nerves is responsible for NO control of the blood vessels.

How well do ecosystem-based planning units represent different componenets of biodiversity?

There are many proposals for managing biodiversity by using surrogates, such as umbrella, indicator, focal, and flagship species. We use the term biodiversity management unit for any ecosystem-based classificatory scheme for managing biodiversity. The sufficiency of biodiversity management unit classification schemes depends upon (1) whether different biotic elements (e.g., trees, birds, reptiles) distinguish between biodiversity management units within a classification (i.e., coherence within classes}; and (2) whether different biotic elements agree upon similarities and dissimilarities among biodiversity management unit classes (i.e., conformance among classes). Recent evaluations suggest that biodiversity surrogates based on few or single taxa are not useful. Ecological vegetation classes are an ecosystem-based classification scheme used as one component for biodiversity management in Victoria, Australia. Here we evaluated the potential for ecological vegetation classes to be used as biodiversity management units in the box-ironbark ecosystem of central Victoria, Australia. Eighty sites distributed among 14 ecological vegetation classes were surveyed in the same ways for tree species, birds, mammals, reptiles, terrestrial invertebrates, and nocturnal flying insects. Habitat structure and geographic separations also were measured, which, with the biotic elements, are collectively referred to as variables. Less than half of the biotic element-ecological vegetation class pairings were coherent. Generalized Mantel tests were used to examine conformance among variables with respect to ecological vegetation classes. While most tests were not significant, birds, mammals, tree species, and habitat structure together showed significant agreement on the rating of similarities among ecological vegetation classes. In this system, use of ecological vegetation classes as biodiversity management units may account reasonably well for birds, mammals, and trees; but reptiles and invertebrates would not be accommodated. We conclude that surrogates will usually have to be augmented or developed as hierarchies to provide general representativeness.

Dual mechanisms for nitric oxide control of large arteries in the estuarine crocodile crocodylus porosus

n reptiles, accumulating evidence suggests that nitric oxide (NO) induces a potent relaxation in the systemic vasculature. However, very few studies have examined the source from which NO is derived. Therefore, the present study used both anatomical and physiological approaches to establish whether NO-mediated vasodilation is via an endothelial or neural NO pathway in the large arteries of the estuarine crocodile Crocodylus porosus. Specific endothelial nitric oxide synthase (NOS) staining was observed in aortic endothelial cells following nicotinamide adenine dinucleotide phosphate-diaphorase (NADPH-d) histochemistry and endothelial NOS immunohistochemistry (IHC), suggesting that an endothelial NO pathway is involved in vascular control. This finding was supported by in vitro organ bath physiology, which demonstrated that the relaxation induced by acetylcholine (10-5 mol l-1) was abolished in the presence of the NOS inhibitor, N-omega-nitro-L-arginine (L-NNA; 10-4 mol l-1), the soluble guanylyl cyclase inhibitor, 1H-[1,2,4]oxadiazolo[4,3-a]quinoxalin-1-one (ODQ; 10-5 mol l-1), or when the endothelium was removed. Interestingly, evidence for a neural NO pathway was also identified in large arteries of the crocodile. Neural NOS was located in perivascular nerves of the major blood vessels following NADPH-d histochemistry and neural NOS IHC and in isolated aortic rings, L-NNA and ODQ, but not the removal of the endothelium, abolished the relaxation effect of the neural NOS agonist, nicotine (3x10-4 mol l-1). Thus, we conclude that the large arteries of C. porosus are potentially regulated by NO-derived from both endothelial and neural NOS.

Regional faunal decline – reptile occurrence in fragmented rural landscapes of south-eastern Australia

Many species of reptiles are sedentary and depend on ground-layer habitats, suggesting that they may be particularly vulnerable to landscape changes that result in isolation or degradation of native vegetation. We investigated patterns of reptile distribution and abundance in remnant woodland across the Victorian Riverina, south-eastern Australia, a bioregion highly modified (>90%) by clearing for agriculture. Reptiles were intensively surveyed by pitfall trapping and censuses at 60 sites, stratified to sample small (<30 ha) and large (>30 ha) remnants, and linear strips of roadside and streamside vegetation, across the regional environmental gradient. The recorded assemblage of 21 species was characterised by low abundance and patchy distribution of species. Reptiles were not recorded by either survey technique at 22% of sites and at a further 10% only a single individual was detected. More than half (53%) of all records were of two widespread, generalist skink species. Multivariate models showed that the distribution of reptiles is influenced by factors operating at several levels. The environmental gradient exerts a strong influence, with increasing species richness and numbers of individuals from east (moister, higher elevation) to west (drier, lower elevation). Differences existed between types of remnants, with roadside vegetation standing out as important; this probably reflects greater structural heterogeneity of ground and shrub strata than in remnants subject to grazing by stock. Although comparative historical data are lacking, we argue that there has been a region-wide decline in the status of reptiles in the Victorian Riverina involving: (1) overall population decline commensurate with loss of >90% of native vegetation; (2) disproportionate decline of grassy dry woodlands and their fauna (cf. floodplains); and (3) changes to populations and assemblages in surviving remnants due to effects of land-use on reptile habitats. Many species now occur as disjunct populations, vulnerable to changing land-use. The status of reptiles in rural Australia warrants greater attention than has been given to date. Effective conservation of this component of the biota requires better understanding of the population dynamics, habitat use and dispersal capacity of species; and a commitment to landscape restoration coupled with effective ecological monitoring.

Testing the immunocompetence handicap hypothesis : a review of the evidence

The immunocompetence handicap hypothesis was formulated 12 years ago in an attempt to offer a proximate mechanism by which female choice of males could be explained by endocrine control of honest signalling. The hypothesis suggested that testosterone has a dual effect in males of controlling the development of sexual signals while causing immunosuppression. Our purpose in this review is to examine the empirical evidence to date that has attempted to test the hypothesis, and to conduct a meta-analysis on two of the assumptions of the hypothesis, that testosterone reduces immunocompetence and increases parasitism, to ascertain any statistical trend in the data. There is some evidence to suggest that testosterone is responsible for the magnitude of trait expression or development of sexual traits, but this is by no means conclusive. The results of many studies attempting to find evidence for the supposed immunosuppressive qualities of testosterone are difficult to interpret since they are observational rather than experimental. Of the experimental studies, the data obtained are ambiguous, and this is reflected in the result of the meta-analysis. Overall, the meta-analysis found a significant suppressive effect of testosterone on immunity, in support of the hypothesis, but this effect disappeared when we controlled for multiple studies on the same species. There was no effect of testosterone on direct measures of immunity, but it did increase ectoparasite abundance in several studies, in particular in reptiles. A funnel analysis indicated that the results were robust to a publication bias. Alternative substances that interact with testosterone, such as glucocorticoids, may be important. Ultimately, a greater understanding is required of the complex relationships that exist both within and between the endocrine and immune systems and their consequences for mate choice decision making.