Interplay between plasma hormone profiles, sex and body condition in immature hawksbill turtles (Eretmochelys imbricata) subjected to a capture stress protocol

We investigated plasma hormone profiles of corticosterone and testosterone in immature hawksbill turtles (Eretmochelys imbricata) in response to a capture stress protocol. Further, we examined whether sex and body condition were covariates associated with variation in the adrenocortical response of immature turtles. Hawksbill turtles responded to the capture stress protocol by significantly increasing plasma levels of corticosterone over a 5 h period. There was no significant sex difference in the corticosterone stress response of immature turtles. Plasma testosterone profiles, while significantly different between the sexes, did not exhibit a significant change during the 5 h capture stress protocol. An index of body condition was not significantly associated with a turtle's capacity to produce plasma corticosterone both prior to and during exposure to the capture stress protocol. In summary, while immature hawksbill turtles exhibited an adrenocortical response to a capture stress protocol, neither their sex nor body condition was responsible for variation in endocrine responses. This lack of interaction between the adrenocortical response and these internal factors suggests that the inactive reproductive- and the current energetic- status of these immature turtles are important factors, that could influence plasma hormone profiles during stress. (C) 2003 Elsevier Inc. All rights reserved.

Automated Analysis of the Auditory Brainstem Response Using Derivative Estimation Wavelets

In this paper, we describe an algorithm that automatically detects and labels peaks I - VII of the normal, suprathreshold auditory brainstem response (ABR). The algorithm proceeds in three stages, with the option of a fourth: ( 1) all candidate peaks and troughs in the ABR waveform are identified using zero crossings of the first derivative, ( 2) peaks I - VII are identified from these candidate peaks based on their latency and morphology, ( 3) if required, peaks II and IV are identified as points of inflection using zero crossings of the second derivative and ( 4) interpeak troughs are identified before peak latencies and amplitudes are measured. The performance of the algorithm was estimated on a set of 240 normal ABR waveforms recorded using a stimulus intensity of 90 dBnHL. When compared to an expert audiologist, the algorithm correctly identified the major ABR peaks ( I, III and V) in 96 - 98% of the waveforms and the minor ABR peaks ( II, IV, VI and VII) in 45 - 83% of waveforms. Whilst peak II was correctly identified in only 83% and peak IV in 77% of waveforms, it was shown that 5% of the peak II identifications and 31% of the peak IV identifications came as a direct result of allowing these peaks to be found as points of inflection. Copyright (C) 2005 S. Karger AG, Basel.

Sox8 is expressed at similar levels in gonads of both sexes during the sex determining period in turtles

A critical gene involved in mammalian sex determination and differentiation is the Sty-related gene Sox9. In reptiles, Sox9 resembles that of mammals in both structure and expression pattern in the developing gonad, but a causal role in male sex determination has not been established. A closely related gene, Sox8, is conserved in human, mouse, and trout and is expressed in developing testes and not developing ovaries in mouse. In this study, we tested the possibility of Sox8 being important for sex determination or sex differentiation in the red-eared slider turtle Trachemys scripta, in which sex is determined by egg incubation temperature between stages 15 and 20. We cloned partial turtle Sox8 and anti-Mullerian hormone (Amh) cDNAs, and analyzed the expression patterns of these genes in developing gonads by reverse transcriptase-polymerase chain reaction and whole-mount in situ hybridization. While Amh is expressed more strongly in males than in females at stage 17, Sox8 is expressed at similar levels in males and females throughout the sex-determining period. These observations suggest that differential transcription of Sill is not responsible for regulation of Amh, nor responsible for sex determination in turtle. (C) 2004 Wiley-Liss, Inc.

Monitoring green turtles (Chelonia mydas) at a coastal foraging area in Baja California, Mexico: multiple indices to describe population status

From June 1995 to August 2002 we assessed green turtle (Chelonia mydas) population structure and survival, and identified human impact, at Bahia de los Angeles, a large bay that was once the site of the greatest sea turtle harvest rates in the Gulf of California, Mexico. Turtles were captured live with entanglement nets and mortality was quantified through stranding surveys and flipper tag recoveries. A total of 14,820 netting hours (617.5 d) resulted in 255 captures of 200 green turtles. Straight-carapace length and mass ranged from 46.0-100.0 cm (mean = 74.3 +/- 0.7 cm) and 14.5-145.0 kg (mean = 61.5 +/- 1.7 kg), respectively. The size-frequency distribution remained stable during all years and among all capture locations. Anthropogenic-derived injuries ranging from missing flippers to boat propeller scars were present in 4% of captured turtles. Remains of 18 turtles were found at dumpsites, nine stranded turtles were encountered in the study area, and flipper tags from seven turtles were recovered. Survival was estimated at 0.58 for juveniles and 0.97 for adults using a joint live-recapture and dead-recovery model (Burnham model). Low survival among juveniles, declining annual catch per unit effort, and the presence of butchered carcasses indicated human activities continue to impact green turtles at this foraging area.

Spatial and temporal variability in somatic growth of green sea turtles (Chelonia mydas) resident in the Hawaiian Archipelago

The somatic growth dynamics of green turtles ( Chelonia mydas) resident in five separate foraging grounds within the Hawaiian Archipelago were assessed using a robust non-parametric regression modelling approach. The foraging grounds range from coral reef habitats at the north-western end of the archipelago, to coastal habitats around the main islands at the southeastern end of the archipelago. Pelagic juveniles recruit to these neritic foraging grounds from ca. 35 cm SCL or 5 kg ( similar to 6 years of age), but grow at foraging-ground-specific rates, which results in quite different size- and age-specific growth rate functions. Growth rates were estimated for the five populations as change in straight carapace length ( cm SCL year) 1) and, for two of the populations, also as change in body mass ( kg year) 1). Expected growth rates varied from ca. 0 - 2.5 cm SCL year) 1, depending on the foraging-ground population, which is indicative of slow growth and decades to sexual maturity, since expected size of first-time nesters is greater than or equal to 80 cm SCL. The expected size- specific growth rate functions for four populations sampled in the southeastern archipelago displayed a non-monotonic function, with an immature growth spurt at ca. 50 - 53 cm SCL ( similar to 18 - 23 kg) or ca. 13 - 19 years of age. The growth spurt for the Midway atoll population in the northwestern archipelago occurs at a much larger size ( ca. 65 cm SCL or 36 kg), because of slower immature growth rates that might be due to a limited food stock and cooler sea surface temperature. Expected age-at-maturity was estimated to be ca. 35 - 40 years for the four populations sampled at the south-eastern end of the archipelago, but it might well be > 50 years for the Midway population. The Hawaiian stock comprises mainly the same mtDNA haplotype, with no differences in mtDNA stock composition between foraging-ground populations, so that the geographic variability in somatic growth rates within the archipelago is more likely due to local environmental factors rather than genetic factors. Significant temporal variability was also evident, with expected growth rates declining over the last 10 - 20 years, while green turtle abundance within the archipelago has increased significantly since the mid-1970s. This inverse relationship between somatic growth rates and population abundance suggests a density-dependent effect on somatic growth dynamics that has also been reported recently for a Caribbean green turtle stock. The Hawaiian green turtle stock is characterised by slow growth rates displaying significant spatial and temporal variation and an immature growth spurt. This is consistent with similar findings for a Great Barrier Reef green turtle stock that also comprises many foraging-ground populations spanning a wide geographic range.

Modelling the effect of fibropapilloma disease on the somatic growth dynamics of Hawaiian green sea turtles

The effect of the tumour-forming disease, fibropapillomatosis, on the somatic growth dynamics of green turtles resident in the Pala'au foraging grounds (Moloka'i, Hawai'i) was evaluated using a Bayesian generalised additive mixed modelling approach. This regression model enabled us to account for fixed effects (fibropapilloma tumour severity), nonlinear covariate functional form (carapace size, sampling year) as well as random effects due to individual heterogeneity and correlation between repeated growth measurements on some turtles. Somatic growth rates were found to be nonlinear functions of carapace size and sampling year but were not a function of low-to-moderate tumour severity. On the other hand, growth rates were significantly lower for turtles with advanced fibropapillomatosis, which suggests a limited or threshold-specific disease effect. However, tumour severity was an increasing function of carapace size-larger turtles tended to have higher tumour severity scores, presumably due to longer exposure of larger (older) turtles to the factors that cause the disease. Hence turtles with advanced fibropapillomatosis tended to be the larger turtles, which confounds size and tumour severity in this study. But somatic growth rates for the Pala'au population have also declined since the mid-1980s (sampling year effect) while disease prevalence and severity increased from the mid-1980s before levelling off by the mid-1990s. It is unlikely that this decline was related to the increasing tumour severity because growth rates have also declined over the last 10-20 years for other green turtle populations resident in Hawaiian waters that have low or no disease prevalence. The declining somatic growth rate trends evident in the Hawaiian stock are more likely a density-dependent effect caused by a dramatic increase in abundance by this once-seriously-depleted stock since the mid-1980s. So despite increasing fibropapillomatosis risk over the last 20 years, only a limited effect on somatic growth dynamics was apparent and the Hawaiian green turtle stock continues to increase in abundance.

Survey-gap analysis in expeditionary research: where do we go from here?

Research expeditions into remote areas to collect biological specimens provide vital information for understanding biodiversity. However, major expeditions to little-known areas are expensive and time consuming, time is short, and well-trained people are difficult to find. In addition, processing the collections and obtaining accurate identifications takes time and money. In order to get the maximum return for the investment, we need to determine the location of the collecting expeditions carefully. In this study we used environmental variables and information on existing collecting localities to help determine the sites of future expeditions. Results from other studies were used to aid in the selection of the environmental variables, including variables relating to temperature, rainfall, lithology and distance between sites. A survey gap analysis tool based on 'ED complementarity' was employed to select the sites that would most likely contribute the most new taxa. The tool does not evaluate how well collected a previously visited site survey site might be; however, collecting effort was estimated based on species accumulation curves. We used the number of collections and/or number of species at each collecting site to eliminate those we deemed poorly collected. Plants, birds, and insects from Guyana were examined using the survey gap analysis tool, and sites for future collecting expeditions were determined. The south-east section of Guyana had virtually no collecting information available. It has been inaccessible for many years for political reasons and as a result, eight of the first ten sites selected were in that area. In order to evaluate the remainder of the country, and because there are no immediate plans by the Government of Guyana to open that area to exploration, that section of the country was not included in the remainder of the study. The range of the ED complementarity values dropped sharply after the first ten sites were selected. For plants, the group for which we had the most records, areas selected included several localities in the Pakaraima Mountains, the border with the south-east, and one site in the north-west. For birds, a moderately collected group, the strongest need was in the north-west followed by the east. Insects had the smallest data set and the largest range of ED complementarity values; the results gave strong emphasis to the southern parts of the country, but most of the locations appeared to be equidistant from one another, most likely because of insufficient data. Results demonstrate that the use of a survey gap analysis tool designed to solve a locational problem using continuous environmental data can help maximize our resources for gathering new information on biodiversity. (c) 2005 The Linnean Society of London.

Patterns of digenean parasitism of bivalves from the Great Barrier Reef and associated waters

Digenean parasites of marine bivalves are relatively poorly known, particularly in Australia. We surveyed 2256 bivalve individuals ( 47 species, 17 families) from Queensland marine waters incorporating south-east Queensland, Heron Island ( southern Great Barrier Reef) and Lizard Island ( northern Great Barrier Reef). Infections of trematode species from three families, Bucephalidae, Gorgoderidae and Monorchiidae, were found. Overall prevalence of infection was 2.3%. The Bucephalidae was the most commonly found family; 11 species were found in Tellinidae, Ostreidae, Isognomonidae and Spondylidae - the latter two previously unknown as hosts for bucephalids. A single gorgoderid infection was found in a venerid, Lioconcha castrensis. Five species of monorchiids were found from Tellinidae and Lucinidae. All infections are new host/parasite records. No infections were found in 35 of the 47 bivalve species sampled. The generally low prevalence of infection by digeneans of bivalves suggests that it is unlikely that any of the species reported here are seriously damaging to bivalve populations in these waters. We deduce that, at best, we have some life-cycle information but no actual identifications for 10% of the species of trematodes that infect bivalves of Queensland marine waters.

Do open-cycle hatcheries relying on tourism conserve sea turtles? Sri Lankan developments and economic-ecological considerations

By combining economic analysis of markets with ecological parameters, this article considers the role that tourism-based sea turtle hatcheries (of an open-cycle type) can play in conserving populations of sea turtles. Background is provided on the nature and development of such hatcheries in Sri Lanka. The modeling facilitates the assessment of the impacts of turtle hatcheries on the conservation of sea turtles and enables the economic and ecological consequences of tourism, based on such hatcheries, to be better appreciated. The results demonstrate that sea turtle hatcheries serving tourists can make a positive contribution to sea turtle conservation, but that their conservation effectiveness depends on the way they are managed. Possible negative effects are also identified. Economic market models are combined with turtle population survival relationships to predict the conservation impact of turtle hatcheries and their consequence for the total economic value obtained from sea turtle populations.

Effects of hatchery shading and nest depth on the development and quality of Chelonia mydas hatchlings: implications for hatchery management in Peninsular, Malaysia

One of the decisions made by hatchery managers around the world is what degree of shading and nest depth are required to maximise the production of high-quality hatchlings at optimal sex ratios. The primary objectives of this study were to determine the effects of (1) hatchery shading and nest depth on nest temperatures and emergence lag, and (2) nest temperatures and nest depth on hatchling sex ratio and quality. In 2001, 26 Chelonia mydas clutches from Ma'Daerah beach, Terengganu, Malaysia, were relocated alternatively at depths of 50 cm and 75 cm into a 70%-shaded and a 100%-shaded hatchery. Data loggers were placed into the centre of each relocated clutch to record the temperature every hour over the course of incubation. When the hatchlings emerged, a sample of the clutch was run, measured and weighed and a separate sample was examined histologically for sex characteristics. Nest temperatures ranged between 28 degrees C and 30 degrees C and generally showed increases over the second half of incubation due to metabolic heating of the clutch. There was no significant correlation found between nest temperature and any of the hatchling parameters measured. Hatchlings from 75-cm-deep nests had a longer emergence lag (46.4 (+/- 10.2) h) than hatchlings from 50-cm-deep nests. Hatch and emergence success were similar to those of natural populations and hatchling sex ratios were male dominant, with an average of 72% males. There was a poor correlation between mean middle-third incubation temperatures and sex ratio. Hatchlings from 75-cm-deep nests had similar running speeds but lower condition index than their conspecifics from 50-cm-deep nests.

Evaluating trends in abundance of immature green turtles, Chelonia mydas, in the Greater Caribbean

Many long-lived marine species exhibit life history traits. that make them more vulnerable to overexploitation. Accurate population trend analysis is essential for development and assessment of management plans for these species. However, because many of these species disperse over large geographic areas, have life stages inaccessible to human surveyors, and/or undergo complex developmental migrations, data on trends in abundance are often available for only one stage of the population, usually breeding adults. The green turtle (Chelonia mydas) is one of these long-lived species for which population trends are based almost exclusively on either numbers of females that emerge to nest or numbers of nests deposited each year on geographically restricted beaches. In this study, we generated estimates of annual abundance for juvenile green turtles at two foraging grounds in the Bahamas based on long-term capture-mark-recapture (CMR) studies at Union Creek (24 years) and Conception Creek (13 years), using a two-stage approach. First, we estimated recapture probabilities from CMR data using the Cormack-Jolly-Seber models in the software program MARK; second, we estimated annual abundance of green turtles. at both study sites using the recapture probabilities in a Horvitz-Thompson type estimation procedure. Green turtle abundance did not change significantly in Conception Creek, but, in Union Creek, green turtle abundance had successive phases of significant increase, significant decrease, and stability. These changes in abundance resulted from changes in immigration, not survival or emigration. The trends in abundance on the foraging grounds did not conform to the significantly increasing trend for the major nesting population at Tortuguero, Costa Rica. This disparity highlights the challenges of assessing population-wide trends of green turtles and other long-lived species. The best approach for monitoring population trends may be a combination of (1) extensive surveys to provide data for large-scale trends in relative population abundance, and (2) intensive surveys, using CMR techniques, to estimate absolute abundance and evaluate the demographic processes' driving the trends.

Cholinergic and adrenergic innervation of lingual salt glands of the estuarine crocodile, Crocodylus porosus

Many marine reptiles and birds possess extrarenal salt glands that facilitate the excretion of excess sodium and chloride ions accumulated as a consequence of living in saline environments. Control of the secretory activity of avian salt glands is under neural control, but little information is available on the control of reptilian salt glands. Innervation of the lingual salt glands of the salt water crocodile, Crocodylus porosus, was examined in salt water-acclimated animals using histological methods. Extensive networks of both cholinergic and adrenergic nerve fibres were identified close to salt-secreting lobules and vasculature. The identification of both catecholamine-containing and cholinergic neurons in the salt gland epithelium and close to major blood vessels in the tissue suggests the action of the neurotransmitters on the salt-secreting epithelium itself and the rich vascular network of the lingual salt glands.

Analysis of the Pasteurella multocida outer membrane sub-proteome and its response to the in vivo environment of the natural host

This study describes the identification of outer membrane proteins (OMPs) of the bacterial pathogen Pasteurella multocida and an analysis of how the expression of these proteins changes during infection of the natural host. We analysed the sarcosine-insoluble membrane fractions, which are highly enriched for OMPs, from bacteria grown under a range of conditions. Initially, the OMP-containing fractions were resolved by 2-DE and the proteins identified by MALDI-TOF MS. In addition, the OMP-containing fractions were separated by 1-D SDS-PAGE and protein identifications were made using nano LC MS/MS. Using these two methods a total of 35 proteins was identified from samples obtained from organisms grown in rich culture medium. Six of the proteins were identified only by 2-DE MALDI-TOF MS, whilst 17 proteins were identified only by 1-D LC MS/MS. We then analysed the OMPs from P. multocida which had been isolated from the bloodstream of infected chickens (a natural host) or grown in iron-depleted medium. Three proteins were found to be significantly up-regulated during growth in vivo and one of these (Pm0803) was also up-regulated during growth in iron-depleted medium. After bioinformatic analysis of the protein matches, it was predicted that over one third of the combined OMPs predicted by the bioinformatics sub-cellular localisation tools PSORTB and Proteome Analyst, had been identified during this study. This is the first comprehensive proteomic analysis of the P. multocida outer membrane and the first proteomic analysis of how a bacterial pathogen modifies its outer membrane proteome during infection.