The primary structure of hemoglobin D from the Aldabra giant tortoise,Geochelone gigantea

The complete primary structures of alpha D-2- and beta-globin of hemoglobin D (Hb D) from the Aldabra giant tortoise, Geochelone gigantea, have been constructed by amino acid sequencing analysis in assistance with nucleotide sequencing analysis of PCR fragments amplified using degenerate oligonucleotide primers. Using computer-assisted sequence comparisons, the alpha D-2-globin shared a 92.0% sequence identity versus alpha D-globin of Geochelone carbonaria, a 75.2% versus alpha D-globin of Aves (Rhea americana) and a 62.4% versus alpha A-globin of Hb A expressed in adult red blood cells of Geochelone gigantea. Additionally, judging from their primary structures, an identical beta-globin was common to the two hemoglobin components, Hb A and Hb D. The alpha D-2- and beta-globin genes contained the three-exon and two-intron configurations and showed the characteristic of all functional vertebrate hemoglobin genes except an abnormal GC dinucleotide instead of the invariant GT at the 5' end of the second intron sequence. The introns of alpha D-2-globin gene were both small (224-bp/first intron, 227-bp/second intron) such that they were quite similar to those of adult alpha-type globins; the beta-globin gene has one small intron (approximately 130-bp) and one large intron (approximately 1590-bp). A phylogenetic tree constructed on primary structures of 7 alpha D-globins from Reptilia (4 species of turtles, 2 species of squamates, and 1 species of sphenodontids) and two embryonic alpha-like globins from Aves (Gullus gullus) and Mammals (Homo sapiens) showed the following results: (1) alpha D-globins except those of squamates were clustered, in which Sphenodon punctatus was a closer species to birds than turtles; (2) separation of the alpha A- and alpha D-globin genes occurred approximately 250 million years ago after the embryonic alpha-type globin-genes (pi' and zeta) first split off from the ancestor of alpha-type globin gene family.     

Crystallization and preliminary X-ray diffraction study of hemoglobin D from the Aldabra giant tortoise, Geochelone gigantea

Hemoglobin D (Hb D) from the Aldabra giant tortoise, Geochelone gigantea, was crystallized by the hanging drop vapor diffusion technique with a precipitant solution containing 10% polyethylene glycol 3350 and 50 mM HEPES-Na, pH 7.5. The Hb D crystals of G. gigantea, which diffract to at least a 2.0 A resolution, belong to the monoclinic space group C2 with unit cell dimensions of a = 112.1 A, b = 62.4 A, c = 54.0 A, and beta = 110.3 degrees. One alphabeta dimer molecule of Hb D existed in an asymmetric unit, with a calculated value of Vm of 2.77 A(3)Da(-1).     

The rise and fall of the Aldabran giant tortoise population.

At the end of the 19th century, after prolonged and extensive harvesting, indigenous giant tortoises had been eliminated from all islands in the Indian Ocean, except Aldabra atoll, where only a few survived. With greatly reduced levels of exploitation during the 20th century, the population recovered to a revised estimated total of 129,000 in 1973-1974, when the first sample census was conducted. A repeat census in 1997 revealed a highly significant reduction in numbers over the past 24 years to an estimated total of 100,000. The great majority of tortoises are still found at relatively high density in south-eastern Grande Terre, where the number of animals has declined by more than one-third. In contrast, low-density subpopulations on Malabar and Picard have almost doubled in size, but they represent less than 5% of the total population. Corroborative evidence for the crash in the Grande Terre subpopulation comes from two independent observations: a significant increase in tortoise mortality; and a significant decline in tortoise counts on long-term population monitoring transects. These population changes are attributed to natural population regulatory mechanisms, exacerbated by low rainfall years in the period 1980-1997, including two consecutive years of below average rainfall in 1995-1996 and 1996-1997.     

The Indian Ocean giant tortoise Testudo gigantea on Aldabra

The Indian Ocean giant tortoise survives on Aldabra, an atoll north-west of Madagascar. The Bristol Seychelles Expedition estimated the total numbers as 33,000 during visits in 1964 and 1965. Growth and movements were recorded during marking experiments. The growth rate with size is plotted, the time scale being derived from the number of rings on the shields of the shell; and the growth curve is compared with that for captive tortoises.
Variation in many characters of supposed taxonomic value is explained as the influence of environmental factors on the normal growth pattern, and of non-adaptive differences expressed during adult life. The present separation of north and south Aldabra, and Seychelles forms is not supported.
An explanation is suggested for the uniformity of shape among tortoises.
Observations were made on the feeding, diurnal movements and mortality of tortoises on Aldabra.     

Hemoglobin-sulfhydryls from tortoise (Geochelone carbonaria) can reduce oxidative damage induced by organic hydroperoxide in erythrocyte membrane

Sulfhydryl groups are important to avoid oxidative damage to the cell. In RBC, tert-butyl hydroperoxide (tert-BOOH) and hydrogen peroxide (H₂O₂) are capable of oxidizing heme and promoting lipid peroxidation. H₂O₂ caused greater oxidation of heme than tert-BOOH, although the oxidation of sulfhydryl groups was similar. Geochelone carbonaria Hb, a rich sulfhydryl protein, inhibited the TBA-reactive substances formation of human erythrocytes exposed to tert-BOOH by about 30%; this decrease was smaller with Geochelone denticulata Hb. Sulfhydryl reagents diminished the number of reactive sulfhydryl groups in the G. carbonaria Hb resulting in a decrease of its antioxidant power, suggesting the involvement of sulfhydryls of Hb in the protection against lipid peroxidation.     

No evidence of contagious yawning in the red-footed tortoise Geochelone carbonaria

Three hypotheses have attempted to explain the phenomenon of contagious yawning.It has been hypothesized that it is a fixed action pattern for which the releasi...     

Pyrogens fail to produce fever in the leopard tortoise Geochelone pardalis

1. The body temperature of seven tortoises, Geochelone pardalis, was measured in a thermal gradient chamber, by indwelling thermocouples, after injection of various pyrogens. 2. The tortoises regulated their body temperature by moving in the chamber. 3. The tortoises did not develop fever in response to any of the pyrogens we tested. 4. The results support the contention that fever in reptiles is not ubiquitous.     

Attachment sites of the tick Amblyomma marmoreum on its tortoise host,Geochelone pardalis

Leopard tortoises (n=162) were collected over a period of 19 months at the National Zoological gardens, Pretoria, South Africa and the attachment sites of larvae, nymphs and adults of A. marmoreum monitored. Most of the larvae (80%) and nymphs (87%) were found on the anterior parts of the tortoise while adults (90%) were found mainly on the posterior regions of the host body. The immature stages were predominant in soft skinned areas protected by the carapace especially the neck and upper leg areas. Male and female ticks were most commonly found on the posterior armpit. posterior legs and the areas surrounding the base of the tail. Interstadial competition as a major factor influencing attachment site segregation is doubtful. Anterior attachment site selection of the immature stages is believed to result largely from ticks lying passively in wait and then climbing onto the host as it moves through the vegetation and the requirement for soft skinned and protected areas to allow for blood feeding. Adult females usually attach posteriorly in areas that allow them to fully engorge without being dislodged by mechanical disturbance, while males prefer to attach at sites where they are protected but still remain in close proximity to female pheromone signals.     

Sighting frequency and food habits of the leopard tortoise, Geochelone pardalis, in northern Tanzania

Sighting frequency and food habits of the leopard tortoise (Geochelone pardalis) were studied in northern Tanzania from October 1993 to June 1996. Sighting frequency varied significantly between protected (0.22 mhr^?1) and unprotected sites (0.59 mhr^?1), and between Arusha (0.27 mhr^?1) and Serengeti sites (0.56 mhr^?1). The tortoise diet comprised mostly plants (97.8%) and rarely inorganic matter (2.2%). A total of 47 plant species from 21 families was eaten. Forbs made up 74.5% of the plant items and monocots the remainder. Succulents contributed 51.0% of the plant items eaten while grasses and legumes accounted for 16.8% and 13.5%, respectively. Some plant items were eaten more frequently than their occurrence in the habitats.     

Growth and carapacial colour variation of the leopard tortoise, Geochelone pardalis babcocki, in northern Tanzania

Observations were made of growth rate and variation in colour pattern of the leopard tortoise ( Geochelone pardalis babcocki ) in northern Tanzania between October 1993 and June 1996. Growth rate differed significantly between tortoises, with immature animals (6.9 ± 1.4 mm month⁻¹) and females (5.7 ± 1.3 mm month⁻¹) exhibiting higher rates of growth than adult animals (2.9 ± 0.8 mm month⁻¹) and males (3.2 ± 1.1 mm month⁻¹), respectively. The faster growth rate in immature animals suggested that growth rate declined with age. The difference in growth rates between the sexes was ascribed to sexual dimorphism, adult females being larger and therefore growing faster than males.
The carapacial colour pattern observed in the northern Tanzanian tortoises was generally similar to that described elsewhere in the species' range. The colour pattern varied between Arusha and Serengeti District tortoises. Animals at the Arusha sites tended to be sparsely coloured (55.6%) compared to the Serengeti tortoises, which had a buff colouration with radiating spots (89.3%).; The sparse colouration provides effective camouflage in the drier Arusha habitats, as does the buff colouration with black spots in the wetter, darker habitats in the Serengeti.     

Plant dispersal by motor cars

Mud from a car driving more than 15 000 km in the growing season of 1986 in the area surrounding Göttingen (FRG) was sampled systematically to assess the size and nature of the car-borne flora. The sludge from front and back mudguards, wheels and other lower parts of the car was set out for germination in a greenhouse. Until the end of 1987, 124 plant species with a total of 3926 seedlings were identified and counted. Germination success was significantly related to the sampling date, the front or back position of the car and the influence of low temperature during exposure. The nature of this potentially car-dispersed flora was compared with the local flora as a whole as well as the flora and plant communities of road verges in the studied area. It can be supposed that almost all plant species growing in plant communities along roadsides can be carried by cars and therefore belong to the car-dispersed flora.     

Plant dispersal, neighbourhood size and isolation by distance

A theoretical relationship between isolation by distance or spatial genetic structure (SGS) and seed and pollen dispersal is tested using extensive spatial-temporal simulations. Although for animals Wright's neighbourhood size N(e) = 4pisigma(2)(t) has been ascertained also, where sigma(2)(t) is the axial variance of distances between parents and offspring, and it was recently confirmed that N(e) = 4pi(sigma(2)(f) + sigma(2)(m))/2 when dispersal of females and males differ, the situation for plants had not been established. This article shows that for a very wide range of conditions, neighbourhood size defined by Crawford's formula N(e) = 4pi(sigma(2)(s) + sigma(2)(p)/2) fully determines SGS, even when dispersal variances of seed (sigma(2)(s)) and pollen sigma(2)(p)) differ strongly. Further, self-fertilization with rate s acts as zero-distance pollen dispersal, and N(e) = 4pi[sigma(2)(s) + sigma(2)(p)(1 - s)/2] fully determines SGS, for most cases where there are both likely parameter values and substantial SGS. Moreover, for most cases, there is a loglinear relationship, I(1) = 0.587 - 0.117 ln(N(e)), between SGS, as measured by I(1), Moran's coefficient for adjacent individuals, and N(e). However, there are several biologically significant exceptions, namely for very low or large N(e), SGS exceeds the loglinear values. There are also important exceptions to Crawford's formula. First, plants with low seed dispersal, high outcross pollen dispersal and high selfing rate show larger SGS than predicted. Second, in plants with very low (near zero) seed dispersal, selfing decreases SGS, opposite expectations. Finally, in some cases seed dispersal is more critical than pollen dispersal, in a manner inconsistent with Crawford's formula.     

When genes meet nomenclature: tortoise phylogeny and the shifting generic concepts of Testudo and Geochelone

We used a five-gene data set (mtDNA: 12S rRNA, 16S rRNA, cyt-b; nDNA: Cmos, Rag2) comprising approximately two-thirds of all extant testudinid species and, for the first time, including all five Testudo species to investigate the question of whether all western Palaearctic testudinids are monophyletic. Further, we examined whether the recently suggested allocation of the African Geochelone pardalis in the otherwise exclusively South African genus Psammobates and of the Malagasy G. yniphora in the monotypic genus Angonoka is justified in the face of considerable morphological evidence against such placements. Our phylogenetic analyses do not support the paraphyly and generic break-up of Testudo, as suggested by previous papers using a smaller taxon sampling and mtDNA data only. We propose a continued usage of the generic name Testudo for all five western Palaearctic tortoise species. Within Testudo, two monophyletic subclades are present, one containing T. hermanni+T. horsfieldii, and the other comprising (T. kleinmanni+T. marginata)+T. graeca. Nomenclaturally, we demonstrate that Eurotestudo Lapparent de Broin et al., 2006, which was recently erected with the type species T. hermanni, is an objective junior synonym of Chersine Merrem, 1820 and Medaestia Wussow, 1916. Recognition of a monotypic genus Angonoka for G. yniphora is unwarranted according to both our re-analysis of sequence data and morphological data. Acknowledging the strong morphological similarity between G. yniphora and G. radiata, we suggest placing both species into the genus Astrochelys. Although sequence data for only one of the three Psammobates species was available for analysis, there is currently no cause to challenge the monophyly of this genus as established on the basis of morphological evidence. Thus, we hypothesize that G. pardalis is sister to a monophyletic Psammobates. In light of the clear morphological gap between G. pardalis and Psammobates species, the recognition of a distinct genus Stigmochelys for the former seems justified.     

The use of classical and operant conditioning in training Aldabra tortoises (Geochelone gigantea), for venipuncture and other,husbandry issues

A variety of nonhuman animals in zoo and research settings have been the subjects of classical and operant conditioning techniques. Much of the published work has focused on mammals, husbandry training, and veterinary issues. However, several zoos are training reptiles and birds for similar procedures, but there has been little of this work published. Using positive reinforcement techniques enabled the training of 2 male and 2 female Aldabra tortoises (Geochelone gigantea) to approach a target, hold steady on target, and stretch and hold for venipuncture. This article discusses training techniques, venipuncture sight, and future training.     

Seasonal photosynthetic gas exchange and water-use efficiency in a constitutive CAM plant, the giant saguaro cactus (Carnegiea gigantea)

Crassulacean acid metabolism (CAM) and the capacity to store large quantities of water are thought to confer high water use efficiency (WUE) and survival of succulent plants in warm desert environments. Yet the highly variable precipitation, temperature and humidity conditions in these environments likely have unique impacts on underlying processes regulating photosynthetic gas exchange and WUE, limiting our ability to predict growth and survival responses of desert CAM plants to climate change. We monitored net CO₂ assimilation (A _net), stomatal conductance (g _s), and transpiration (E) rates periodically over 2 years in a natural population of the giant columnar cactus Carnegiea gigantea (saguaro) near Tucson, Arizona USA to investigate environmental and physiological controls over carbon gain and water loss in this ecologically important plant. We hypothesized that seasonal changes in daily integrated water use efficiency (WUE_day) in this constitutive CAM species would be driven largely by stomatal regulation of nighttime transpiration and CO₂ uptake responding to shifts in nighttime air temperature and humidity. The lowest WUE_day occurred during time periods with extreme high and low air vapor pressure deficit (D _a). The diurnal with the highest D _a had low WUE_day due to minimal net carbon gain across the 24 h period. Low WUE_day was also observed under conditions of low D _a; however, it was due to significant transpiration losses. Gas exchange measurements on potted saguaro plants exposed to experimental changes in D _a confirmed the relationship between D _a and g _s. Our results suggest that climatic changes involving shifts in air temperature and humidity will have large impacts on the water and carbon economy of the giant saguaro and potentially other succulent CAM plants of warm desert environments.