绿海龟稚龟的人工饲养

暂养绿海龟稚龟是提高其成活率,保护其种群资源的有效途径。本文阐述了绿海龟稚龟的人工饲养与疾病防治技术。     

Conservation Genetics of the East Pacific Green Turtle (Chelonia mydas) in Michoacan, Mexico

The main continental nesting rookeries of the east Pacific green turtle (EPGT), Chelonia mydas, on the Michoacan (Mexico) coast suffered drastic population declines following intense exploitation in the 1960s--1970s with annual abundance of nesting females plummeting from about 25,000 to an average of about 1400 between 1982 and 2001. Analyses of data from three nDNA microsatellite loci and 400 bp mtDNA control region sequences from a total of 123 nesting females sampled from four Michoacan rookeries found no evidence of population sub-structuring. The recent order of magnitude reduction in the population size shows no apparent impact on genetic diversity in either control region sequences (overall h = 0.48; pi = 0.0036) or microsatellite loci (overall Na = 20.8; Hexp = 0.895). Our estimates of annual effective female population size (Nef; from theta = 2Nemicron) of 1.9-2.3 x 10(3), in spite of being an order of magnitude below historical records, appear to be sufficient to allow recovery of this population without significant loss of genetic diversity. These findings highlight the importance of continued conservation to reverse the decline of this population before it becomes vulnerable to genetic erosion.     

Characterization of polymorphic microsatellite markers for the green turtle (Chelonia mydas)

We describe primers and polymerase chain reaction conditions to amplify 12 microsatellite loci from the green turtle (Chelonia mydas), including one dinucleotide, four trinucleotide and seven tetranucleotide loci. The primers were tested on 78 individuals from a Pacific population nesting in the Hawaiian Islands. The primer pairs developed in this study yielded an average of 8.33 alleles per locus (range of 3-15 alleles), an average observed heterozygosity of 0.668 (range 0.309-0.910), and an average polymorphic information content of 0.647 (range 0.287-0.894).     

Metabolic heating in Mediterranean loggerhead sea turtle clutches

Offspring sex ratio is an important demographic parameter and, given its determination by incubation temperature in sea turtles, might be a key factor for their conservation under climate warming. An appealing approach to estimate hatchling sex ratios is to measure sand temperatures at nest depth and deduce hatchling sex ratios from a beforehand-established relationship of hatchling sex ratio and sand temperature. Such estimates will only be accurate though if metabolic heat produced by the embryos is considered. Judging whether metabolic heating has a potential effect on hatchling sex ratios without actually measuring temperature within clutches would greatly facilitate monitoring protocols. We tested for a relationship between the amount of metabolic heating and the number of developed embryos as well as clutch size in the largest known loggerhead sea turtle (Caretta caretta) population of the Mediterranean on Zakynthos (Greece). Temperatures were measured within 20 nests as well as at a reference site in the sand at nest depth. Metabolic heating was detected, but only during the last third of the incubation period did nests heat up considerably (1.6 °C on average) above the temperature of the surrounding sand. During the middle third of incubation, when sex is determined, the amount of metabolic heating was negligible. The amount of metabolic heating during the last third of the incubation duration was significantly correlated to the number of offspring developed to at least about 75% of incubation duration. This factor explained nearly 50% of variation in metabolic heating. Metabolic heating was also significantly correlated to clutch size. Given that clutch size within the Mediterranean is largest in Zakynthos loggerheads, we conclude that metabolic heating can be ignored in the estimate of hatchling sex ratios in Mediterranean loggerhead populations. These results thus provide the basis for a feasible monitoring of hatchling sex ratios in the loggerhead sea turtle in the Mediterranean.     

Characterization of single nucleotide polymorphism markers for the green sea turtle (Chelonia mydas)

We present data on 29 new single nucleotide polymorphism assays for the green sea turtle, Chelonia mydas. DNA extracts from 39 green turtles were used for two methods of single nucleotide polymorphism discovery. The first approach employed an amplified fragment length polymorphism technique. The second technique screened a microsatellite library. Allele-specific amplification assays were developed for high-throughput single nucleotide polymorphism genotyping and tested on two Pacific C. mydas nesting populations. Observed heterozygosities ranged from 0 to 0.95 for a Hawaiian population and from 0 to 0.85 for a Galapagos population. Each of the populations had one locus out of Hardy-Weinberg equilibrium, SSCM2b and SSCM5 for Hawaii and Galapagos, respectively. No loci showed significant genotypic linkage disequilibrium across an expanded set of four Pacific nesting populations. However, two loci, SSCM4 and SSCM10b showed linkage disequilibrium across three populations indicating possible association.     

Amino Acid Sequence and Activity of Green Turtle (Chelonia mydas) Lysozyme

Green turtle lysozyme purified from egg white was sequenced and analyzed its activity. Lysozyme was reduced and pyridylethylated or carboxymethylated to digest with trypsin, chymotrypsin and V8 protease. The peptides yielded were purified by RP-HPLC and sequenced. Every trypsin peptide was overlapped by chymotrypsin peptides and V8 protease peptides. This lysozyme is composed of 130 amino acids including an insertion of a Gly residue between 47 and 48 residues when compared with chicken lysozyme. The amino acid substitutions were found at subsites E and F. Namely Phe34, Arg45, Thr47, and Arg114 were replaced by Tyr, Tyr, Pro, and Asn, respectively. The time course using N-acetylglucosamine pentamer as a substrate showed a reduction of the rate constant of glycosidic cleavage and transglycosylation and increase of binding free energy for subsite E, which proved the contribution of amino acids mentioned above for substrate binding at subsites E and F.     

The Complete Amino Acid Sequence of Green Turtle (Chelonia mydas) Egg White Ribonuclease

Egg white ribonuclease was first found in green turtle eggs. This enzyme has been purified by CM-toyopearl cation exchange. Two isoforms (GTRNase-1 and GTRNase-2) were further separated by RP-HPLC, with the same M.W. (13 kDa) and activity. These isoforms carried one amino acid exchange of Ser and Leu at the position 37. The N-terminal sequence, ETRYEKF, was determined for the transblotted protein. Internal sequences were analyzed by protein sequencer and ESI-Q-TOF mass spectrometry for tryptic peptides (Ts). The overlapping sequences were obtained from chymotryptic peptides, CNBr fragments and ISD-MS/MS analysis. The C-terminal Ile was identified by CPase-Y. The established sequence composed of 119 residues with the molecular mass of 12,942.1 Da for GTRNase-1 and 12,967.8 Da for GTRNase-2. The comparison of sequence with known pancreatic RNases, 27 positions including catalytic residues at the position 11 and 114 were conserved. Also basic residues contributed to phosphate binding residues were conserved with the exception of Lys 66. One insertion at the position 14, and 3 deletions at the position-1, between position 64–65, and 110 and 111 were found. Two Cys residues at position 65 and 72 that form a disulfide bond in mammalian RNase were deleted and exchanged. All these difference in the sequence were similar to reptile pancreatic RNase.Data deposition: The sequence reported in this paper has been submitted to the UniProt Knowledgebase under accession No. P84844.     

Phylogeography of the green turtle, Chelonia mydas, in the Southwest Indian Ocean

Patterns of mitochondrial DNA (mtDNA) variation were used to analyse the population genetic structure of southwestern Indian Ocean green turtle (Chelonia mydas) populations. Analysis of sequence variation over 396 bp of the mtDNA control region revealed seven haplotypes among 288 individuals from 10 nesting sites in the Southwest Indian Ocean. This is the first time that Atlantic Ocean haplotypes have been recorded among any Indo-Pacific nesting populations. Previous studies indicated that the Cape of Good Hope was a major biogeographical barrier between the Atlantic and Indian Oceans because evidence for gene flow in the last 1.5 million years has yet to emerge. This study, by sampling localities adjacent to this barrier, demonstrates that recent gene flow has occurred from the Atlantic Ocean into the Indian Ocean via the Cape of Good Hope. We also found compelling genetic evidence that green turtles nesting at the rookeries of the South Mozambique Channel (SMC) and those nesting in the North Mozambique Channel (NMC) belong to separate genetic stocks. Furthermore, the SMC could be subdivided in two different genetic stocks, one in Europa and the other one in Juan de Nova. We suggest that this particular genetic pattern along the Mozambique Channel is attributable to a recent colonization from the Atlantic Ocean and is maintained by oceanic conditions in the northern and southern Mozambique Channel that influence early stages in the green turtle life cycle.     

Long-distance migration and homing after displacement in the green turtle (Chelonia mydas): a satellite tracking study

Four green turtle females were tracked by satellite during their post-reproductive migration in the South China Sea. Three of them reached their feeding grounds 923–1551 km distant. During nesting activity, a female was displaced twice, and her return trips to the nesting beach from 11 and 284 km were tracked by a direction-recording data-logger and by satellite, respectively. Part of the journeys occurred coastwise, indicating that leading geographical features had been utilised. The straightness of the turtles' tracks in open seas, both over shallow and deep waters, and their ability to pinpoint distant targets and home after displacement off their usual routes, provides circumstantial evidence for a true navigation mechanism.Abbreviation PTT platform transmitter terminal     

Structure and Function of the Gastrointestinal Tract of the Green Turtle(Chelonia mydas) Hatchling

The gastrointestinal tracts of four Chelonia mydas hatchlings were examined at the anatomical, histological and ultrastructural level. Our results show that the gastrointestinal tract(GI) is composed by esophagus, stomach, small intestine(SI) and large intestine(LI), and histologically of mucosa, submucosa, muscularis externa(ME) and serosa. The esophagus is marked by conical papillae lined by keratinized stratified squamous epithelium, whereas the remaining GI by simple columnar epithelium; esophageal diverticulum is absent. The stomach covered with mucous granule cells, contains cardia, fundic regions and pylorus, which are separately characterized by cardiac glands, fundic glands and pyloric glands, and have the thickest submucosa and ME of the GI. The ME of the esophagus mainly consist of one layer of circular smooth muscle whereas the rest of GI of two layers, inner circular muscle and outer longitudinal muscle. The SI is slightly longer than the LI and the GI is approximately 5.11 times of the carapace length. The SI is lined with longitudinal zigzag folds and characterized by absorptive cells with longer and denser microvilli, whereas the LI by transversal folds, goblet cells and lymphoid nodules. Only intestinal glands appear in duodenum. Endocrine cells are observed in all sections of the GI and accounted for the largest proportion in duodenum. The results demonstrate a perfect combination of the structure and function of the GI and reveal that the digestion and absorption primarily occurs in the foregut. C. mydas hatchling may prefer carnivorous diet.     

Sand and nest temperatures and an estimate of hatchling sex ratio from the Heron Island green turtle (Chelonia mydas) rookery, Southern Great Barrier Reef

Sand and nest temperatures were monitored during the 2002–2003 nesting season of the green turtle, Chelonia mydas, at Heron Island, Great Barrier Reef, Australia. Sand temperatures increased from ∼ 24°C early in the season to 27–29°C in the middle, before decreasing again. Beach orientation affected sand temperature at nest depth throughout the season; the north facing beach remained 0.7°C warmer than the east, which was 0.9°C warmer than the south, but monitored nest temperatures were similar across all beaches. Sand temperature at 100 cm depth was cooler than at 40 cm early in the season, but this reversed at the end. Nest temperatures increased 2–4°C above sand temperatures during the later half of incubation due to metabolic heating. Hatchling sex ratio inferred from nest temperature profiles indicated a strong female bias.     

The influence of diet on fatty acids in the egg yolk of green sea turtles, Chelonia mydas

Fatty acid concentrations found in the yolk of green sea turtles reflect differences in the diet of the mothers. All of the 12 fatty acids measured in yolk samples were significantly different between eggs produced from the pellet and wild-type diets. However, the relative pattern of yolk fatty acids in the green turtle mirrored those of other reptiles. Yolk samples contained mostly (63–67%) 14:0. 16:0, 16:1n-7 and 18:1n-9. Yolks from captive animals on pellet diet contained an additional 17.64% of the total yolk lipid as 12:0 and 18:2n-6. Wild yolks contained an extra 11.41% of lipid as 18:0 and 18:1n-7. Selection of fatty acids for the yolk should balance the energetic and anabolic needs of the embryo. Eggs are provisioned based on maternal metabolism of available nutrients and subtle differences between natural foods and those available in captivity could affect the viability of future eggs.     

The effect of organochlorines and heavy metals on sex steroid-binding proteins in vitro in the plasma of nesting green turtles, Chelonia mydas

In this study on green turtles, Chelonia mydas, from Peninsular Malaysia, the effect of selected environmental toxicants was examined in vitro. Emphasis was placed on purported hormone-mimicking chemicals such as dichlorodiphenyltrichloroethane (DDT), dichlorodiphenyldichloroethylene, dieldrin, lead, zinc and copper. Five concentrations were used: high (1 mg/L), medium (10⁻¹ mg/L), low (10⁻² mg/L), very low (10⁻⁶ mg/L) and control (diluted carrier solvent but no toxicants). The results suggest that environmental pesticides and heavy metals may significantly alter the binding of steroids [i.e. testosterone (T) and oestradiol] to the plasma proteins in vitro. Competition studies showed that only Cu competed for binding sites with testosterone in the plasma collected from nesting C. mydas. Dieldrin and all heavy metals competed with oestradiol for binding sites. Furthermore, testosterone binding affinity was affected at various DDT concentrations and was hypothesised that DDT in vivo may act to inhibit steroid-protein interactions in nesting C. mydas. Although the precise molecular mechanism is yet to be described, DDT could have an effect upon the protein conformation thus affecting T binding (e.g. the T binding site on the steroid hormone binding protein molecule).     

Relationship between the major phosphorylated metabolic intermediates and oxygen affinity of whole blood in the loggerhead (Caretta caretta) and the Green Sea Turtle (Chelonia mydas) during development.

(1) 2,3-Diphosphoglyceric acid (2,3-DPG) is present in the erythrocytes (RBC) of the 68-day loggerhead turtle embryo and 44-day green sea turtle embryo at levels of 7.4 and 5.5 μmoles/ml of RBC, representing the major organic phosphate during the latter period of embryonic development. (2) Inositol pentaphosphate (IPP) is absent in the red blood cells of the embryos of both the loggerhead and green sea turtle. (3) Near equimolar amounts of 2,3-DPG and IPP are present in the erythrocytes of the adult loggerhead and green sea turtle. The total concentration of these two organic phosphates is approximately 0.75 μmoles/ml of RBC in the adult of both species. (4) There is a switch from embryonic to adult hemoglobin during development of these two species of turtles; the two embryonic bands have identical electrophoretic mobilities, whereas the two adult bands migrate differently on cellulose acetate at pH 8.6. (5) The whole blood oxygen affinity of the adult loggerhead and green sea turtle is 60.3 and 32.6 Torr, respectively. (6) The stripped adult hemoglobins in these two species of turtles show no change in oxygen affinity upon addition of 2,3-DPG, ATP, or IPP. (7) It therefore appears unlikely that whole blood oxygen affinity is controlled by organic phosphate modulation of hemoglobin function in these species of turtles.     

Green turtles (Chelonia mydas) foraging at Arvoredo Island in Southern Brazil: Genetic characterization and mixed stock analysis through mtDNA control region haplotypes

We analyzed mtDNA control region sequences of green turtles (Chelonia mydas) from Arvoredo Island, a foraging ground in southern Brazil, and identified eight haplotypes. Of these, CM-A8 (64%) and CM-A5 (22%) were dominant, the remainder presenting low frequencies (< 5%). Haplotype (h) and nucleotide (π) diversities were 0.5570 ± 0.0697 and 0.0021 ± 0.0016, respectively. Exact tests of differentiation and AMOVA Φ(ST) pairwise values between the study area and eight other Atlantic foraging grounds revealed significant differences in most areas, except Ubatuba and Rocas/Noronha, in Brazil (p > 0.05). Mixed Stock Analysis, incorporating eleven Atlantic and one Mediterranean rookery as possible sources of individuals, indicated Ascension and Aves islands as the main contributing stocks to the Arvoredo aggregation (68.01% and 22.96%, respectively). These results demonstrate the extensive relationships between Arvoredo Island and other Atlantic foraging and breeding areas. Such an understanding provides a framework for establishing adequate management and conservation strategies for this endangered species.     

The effect of bat (Rousettus aegyptiacus) dispersal on seed germination in eastern Mediterranean habitats

The fruit-bat Rousettus aegyptiacus (Pteropodidae) in Israel consumes a variety of cultivated and wild fruits. The aim of this study was to explore some of its qualities as a dispersal agent for six fruit-bearing plant species. The feeding roosts of the fruit-bat are located an average of 30 m from its feeding trees and thus the bats disperse the seeds away from the shade of the parent canopy. The bat spits out large seeds but may pass some (2%) of the small seeds (<4 mg) through its digestive tract. However, neither the deposited seeds nor the ejected seeds (except in one case) had a significantly higher percentage germinating than intact seeds. Although the fruit-bat did not increase the percentage germinating, seeds of three plant species subject to different feeding behaviors (deposited in feces or spat out as ejecta) had a different temporal pattern of germination from the intact seeds. The combined seed germination distribution generated by these different treatments is more even over time than for each treatment alone. It is sugested that this increases asynchronous germination and therefore enhances plant fitness by spreading the risks encountered during germination, especially in eastern Mediterranean habitats where the pattern of rainfall is unpredictable.     

Mitochondrial DNA polymorphism and feminizing sex factors dynamics in a natural population of Armadillidium vulgare (Crustacea,Isopoda)

Sex determination in Armadillidium vulgare may be under the control of two parasitic sex factors that reverse genetic males into functional neo-females. The first feminizing factor (F) is a Wolbachia and the other (f) is probably a sequence of the F bacterial DNA unstably integrated into the host genome. Both of these feminizing factors are mainly maternally transmitted. Here we investigate the mitochondrial DNA polymorphism of wild iso-female lineages harbouring either F or f. Among the four haplotypes present in the population, two were the f-harbouring lineages, while two were common to the F- and f-harbouring lineages. This result suggests that there has been an introgression of the f factor into lineages infected by F Wolbachia. Based on previous data, we propose two different ways to account for such introgression. Given the particular dynamics of feminizing factors (f-harbouring lineages increase in populations at the expense of F-harbouring lineages), such an introgression should prevent the replacement of F-linked mitochondrial types by f-linked mitochondrial types in wild populations.     

Green turtle (Chelonia mydas) foraging and nesting aggregations in the Caribbean and Atlantic: impact of currents and behavior on dispersal

Although significant amounts of research have been dedicated to increasing the knowledge of the life history of green turtles (Chelonia mydas), large gaps exist in our understanding of juvenile migratory behavior. These gaps can be filled by genetic studies of foraging ground aggregations. Using mitochondrial DNA markers and Bayesian analyses, samples (n = 106) from a foraging aggregation in North Carolina indicated that animals from the east coast of the United States (54%) and Mexico (27%) dominate the composition with the remainder coming from other Caribbean and Atlantic nesting aggregations. These findings prompted a reanalysis of 4 regional foraging aggregations using Bayesian mixed stock analysis, analysis of molecular variance, and diversity measures. Significant regional population structure between northern and southern foraging aggregations in the Caribbean was detected (phiST = 0.27, P = 0.000) in addition to significant nesting aggregation structure (phiST = 0.87, P = 0.000). Haplotype diversity levels were highest at foraging aggregations located within the confluence of major current systems. These findings indicate that both currents and behavior have strong influences on the composition of foraging aggregations. In addition, our results provide evidence of juvenile homing to regional foraging grounds and highlight the difficulties of separating historical and current effects on recruitment patterns at foraging locations.     

The littoral fish community of the Lebanese rocky coast (eastern Mediterranean Sea) with emphasis on Red Sea immigrants

The organisation of the coastal rocky fish community of Lebanon was investigated for the first time, using visual censuses. A total of 62 fish species were recorded between the surface and 32 m depth, 8 species being Red Sea migrants. Species richness and fish abundance were positively correlated first with substrate complexity and second with depth. The trophic structure of the community was dominated by diurnal zooplanktivores (Chromis chromis, Oblada melanura and Spicara smaris) and mesocarnivores 1 (Coris julis and Thalassoma pavo), and did not vary significantly with depth. The Lessepsian migrants represented 13% of the species richness and 19% of the total abundance of individuals. They dominated among herbivores, nocturnal zooplanktivores and macrocarnivores. The Lessepsian Siganus luridus and S. rivulatus (Siganidae), Pempheris vanicolensis (Pempheridae) and Sargocentron rubrum (Holocentridae) were now among the most common fish species on the Lebanese rocky coast. Lessepsian fish species displayed either similar ecological niches in the Red Sea and in the Levantine basin (P. vanicolensis, S. rubrum) or an enlargement of their depth distribution towards deeper waters in their new environment (S. luridus, S. rivulatus). This study will serve as a baseline for future studies as eastern Mediterranean communities are subjected to a steady increase in Red Sea migrant species.