Hematological profile of Chelonia mydas (Testudines, Cheloniidae) according to the severity of fibropapillomatosis or its absence

The green turtle Chelonia mydas feeds and nests in the Brazilian coastal area and is considered an endangered species by the World Conservation Union (IUCN 2009) and threatened by the Red List of Brazilian Fauna (Ministério do Meio Ambiente 2009). Fibropapillomatosis is a disease characterized by benign skin tumors (fibropapillomas), and it is one of the main threats to the survival of this species. Studies suggest the involvement of viruses as infectious agents associated with environmental and genetic factors. Blood samples were collected from 45 turtles captured in the coastal area of the state of Sao Paulo, Brazil. From these, 27 were affected by fibropapillomas and 18 were tumor free. Biometrical data on the turtles, size, location and quantity of tumors were recorded. The area occupied by fibropapillomas per animal was calculated and four groups were determined according to severity of the disease or its absence. The objective of the study was to compare hemogram results of the sea turtles classified in these four groups. The lowest hematocrit value was observed in severely affected animals. In the hemoglobin assay, the highest value was observed in the group of tumor free turtles and the lowest, in animals severely affected. Lymphocyte counts and curved carapace length were on the verge of statistical significance.

Detection of polymorphisms of the mtDNA control region of Caretta caretta (Testudines: Cheloniidae) by PCR-SSCP

Marine turtles are increasingly being threatened worldwide by anthropogenic activities. Better understanding of their life cycle, behavior and population structure is imperative for the design of adequate conservation strategies. The mtDNA control region is a fast-evolving matrilineal marker that has been employed in the study of marine turtle populations. We developed and tested a simple molecular tracing system for Caretta caretta mtDNA haplotypes by polymerase chain reaction-single strand conformation polymorphism (PCR-SSCP). Using this technique, we were able to distinguish the SSCP patterns of 18 individuals of the haplotypes CC-A4, CC-A24 and CCxLO, which are commonly found in turtles sampled on the Brazilian coast. When we analyzed 15 turtles with previously unknown sequences, we detected two other haplotypes, in addition to the other four. Based on DNA sequencing, they were identified as the CC-A17 and CC-A1 haplotypes. Further analyses were made with the sea turtles, Chelonia mydas (N = 8), Lepidochelys olivacea (N = 3) and Eretmochelys imbricata (N = 1), demonstrating that the PCR-SSCP technique is able to distinguish intra-and interspecific variation in the family Cheloniidae. We found that this technique can be useful for identifying sea turtle mtDNA haplotypes, reducing the need for sequencing.

Heavy Metal Contamination in Phrynops geoffroanus (Schweigger, 1812) (Testudines: Chelidae) in a River Basin, So Paulo, Brazil

The Piracicaba River basin is considered the most disturbed river basin in the state of So Paulo. Considerable amounts of agricultural residues are seasonally drained into the river, and the region is also highly urbanized and industrialized with an incipient sewage treatment system. The presence of heavy metals has been previously reported for the water and riverbed in Piracicaba river basin. In this study we evaluated 13 heavy metals in the blood of 37 Geoffroy`s side-necked turtles, Phrynops geoffroanus, from Piracicaba River and Piracicamirim Creek, one of its tributaries. Blood levels of As, Co, Cr, Se and Pb varied among sites, whereas Sn varied between males and females. However, no obvious pathology was detected. Serum level of Cu (2,194 ng g(-1)) and Pb (1,150 ng g(-1)) found in this study are the highest ever described for any reptile; however, no clinical symptoms have been detected in the present study. There is no information about the time scale of such contamination, which could be currently subclinical and yet lead to a breakdown in the population reproductive success in a few years. Based on the present study, legal enforcement is urged in order to locate and extirpate heavy metal sources in the Piracicaba River basin. In addition, monitoring should include humans and commercial fish consumed in local markets.

Structure of a Population of Hydromedusa maximiliani (Testudines, Chelidae) from Parque Estadual da Serra do Mar, an Atlantic Rainforest Preserve in Southeastern Brazil

We studied the structure of a population of Hydromedusa maximiliani associated with a stream in Parque Estadual da Serra do Mar, Nucleo Itutinga-Piloes, southeastern Brazil, between October 2004 and October 2005. Twenty-five individuals were captured, and a population size of 43.72 +/- 23.7 individuals was estimated. This value is similar to that of the population of Parque Estadual Carlos Botelho, another Atlantic forest reserve of southeastern Brazil. Males were recaptured more frequently than females, suggesting higher activity and/or greater movement of males.

Repeated disturbances and canopy disturbance regime in a tropical semi-deciduous forest

The canopy disturbance regime and the influence of gap methods on the interpretation of forest structure and dynamics were evaluated in a tropical semi-deciduous forest in south-eastern Brazil. We encountered a gap density of 11.2 gaps ha(-1) and an average size which varied from 121 to 333 m(2) depending on the gap delimitation method considered (minimum gap size was 10 m(2)). Although average size was slightly higher, the median value obtained (78 m(2)) was comparable to other tropical forest sites and the gap size-class distribution found supported the pattern described for such forest sites. Among 297 gap makers, snapping and uprooting were the most common modes of disturbance. The number and basal area of gap makers were good predictors of gap size. Almost 25% of all gaps suffered from repeated disturbance events that brought about larger gap sizes. Such processes, along with delimitation methods, strongly influenced the estimation of turnover rate and therefore the interpretation of forest dynamics. These results demonstrated the importance of further studies on repeated disturbances, which is often neglected in forest studies.

Macrogeographic Genetic Variation in Broad-Snouted Caiman (Caiman latirostris)

Broad-snouted caiman`s (Caiman latirostris) geographic distribution comprises one of the widest latitudinal ranges among all crocodilians. In this study we analyzed the relationship between geographic distance (along the species latitudinal range) and genetic differentiation using DNA microsatellite loci developed for C. latirostris and Alligator mississippiensis. The results suggest that there is a consistent relationship between geographic distance and genetic differentiation; however, other biogeographical factors seem to be relevant. The Atlantic Chain (Serra do Mar) seems to be an effective geographic barrier, as well as the relatively narrow (<= 1.5 km) sea channel between Cardoso Island and the continent. In addition, coastal populations seem to have been well connected in recent geological time (Pleistocene 16,000 years ago) all along the eastern Brazilian coast. Further studies should focus on the Sao Francisco River drainage, which is still poorly known for this species. J. Exp. Zool. 30.9A:628-636, 2008. (C) 2008 Wiley-Liss, Inc.

The Heart and Patterns of Cardiac Outflow in Crocodilia

The anatomy of the crocodilian heart and major arteries has fascinated people for a very long time. The first scientific paper seems to be that by the Italian anatomist Bartolomeo Panizza in 1833 who wrote about the structure of the heart and the circulation of the blood in /Crocodilys lucius/, an early name for the American Alligator. Since 1833 there have been many papers and the crocodilian heart has attracted the attention of generation after generation of anatomists and physiologists with ever-increasingly sophisticated investigatory techniques be­ing applied to questions about the functional significance of the puzzlingly complex anatomy.

Lesions caused by cardiovascular flukes (Digenea : Spirorchidae) in stranded green turtles (Chelonia mydas)

Evidence of infection with spirorchid flukes (Digenea: Spirorchidae) was sought at necropsy of 96 stranded green turtles, Chelonia mydas, that were examined during the course of a survey of marine turtle mortality in southeastern Queensland, Australia. Three species of spirorchid (Hapalotrema mehrai, H. postorchis, and Neospirorchis schistosomatoides) were identified. Severe disease due to spirorchid fluke infection (spirorchidiasis) was implicated as the principal cause of mortality in 10 turtles (10%), and appeared to be one of multiple severe problems in an additional 29 turtles (30%). Although flukes were observed in only 45% of stranded C. mydas in this study, presumed spirorchid fluke infection was diagnosed in an additional 53% of turtles, based principally on characteristic necropsy lesions and to a lesser extent on the histopathological detection of spirorchid eggs. Characteristic necropsy lesions included miliary spirorchid egg granulomas, which were observed most readily on serosal surfaces, particularly of the small intestine. Cardiovascular lesions included mural endocarditis, arteritis, and thrombosis, frequently accompanied by aneurysm formation. Resolution of thrombi was observed to occur via a combination of granuloma formation about indigestible components (spirorchid fluke egg shells) and exteriorization through the vessel wall, which resulted in granulomatous nodules on the adventitial surface. Septic aortic thrombosis complicated by disseminated bacterial infection, observed in five turtles, was recorded for the first time. Egg granulomas were ubiquitous in turtle tissues throughout this study. Although they generally appeared to be mild or incidental lesions, they were occasionally associated with severe multifocal granulomatous pneumonia or meningitis.

Glossocercus chelodinae (MacCallum, 1921) n. comb. (Cestoda : Dilepididae) from freshwater turtles in Australia and a redefinition of the genus Bancroftiella Johnston, 1911

Glossocercus chelodinae (MacCallum, 1921) n. comb. is redescribed from fresh material recovered from the intestine of an Australian freshwater turtle, Chelodina expansa. G. chelodinae can be distinguished from all other species of the genus by the shape of its rostellar hooks. it is suggested that this species has colonised fish-eating turtles from fish-eating birds. The morphological relationships among Parvitaenia, Bancroftiella and Glossocercus are discussed. The diagnosis of Bancroftiella is amended and marsupials are eliminated as hosts. Bancroftiella sudarikovi Spasskii & Yurpalova, 1970 becomes a synonym of Glossocercus glandularis (Fuhrmann, 1905); only B. tennis Johnston, 1911, the type-species, and B. ardeae Johnston, 1911 remain in the genus.

Peptidergic control of gastrointestinal blood flow in the estuarine crocodile, Crocodylus porosus

Peptidergic mechanisms influencing the resistance of the gastrointestinal vascular bed of the estuarine crocodile, Crocodylus porosus, were investigated. The gut was perfused in situ via the mesenteric and the celiac arteries, and the effects of different neuropeptides were tested using bolus injections. Effects on vascular resistance were recorded as changes in inflow pressures. Peptides found in sensory neurons [substance P, neurokinin A, and calcitonin gene-related peptide (CGRP)] all caused significant relaxation of the celiac vascular bed, as did vasoactive intestinal polypeptide (VIP), another well-known vasodilator. Except for VIP, the peptides also induced transitory gut contractions. Somatostatin and neuropeptide Y (NPY), which coexist in adrenergic neurons of the C. porosus, induced vasoconstriction in the celiac vascular bed without affecting the gut motility. Galanin caused vasoconstriction and occasionally activated the gut wall. To elucidate direct effects on individual vessels, the different peptides were tested on isolated ring preparations of the mesenteric and celiac arteries. Only CGRP and VIP relaxed the epinephrine-precontracted celiac artery, whereas the effects on the mesenteric artery were variable. Somatostatin and NPY did not affect the resting tonus of these vessels, but somatostatin potentiated the epinephrine-induced contraction of the celiac artery. Immunohistochemistry revealed the existence and localization of the above-mentioned peptides in nerve fibers innervating vessels of different sizes in the gut region. These data support the hypothesis of an important role for neuropeptides in the control of the vascular bed of the gastrointestinal tract in C. porosus.

Relationships between blood pressure and heart rate in the saltwater crocodile Crocodylus porosus

The cardiac limb of the baroreflex loop was studied in the saltwater crocodile Crocodylus porosus, The classical pharmacological methodology using phenylephrine and sodium nitroprusside was used to trigger blood pressure changes, and the resulting alterations in heart rate were analysed quantitatively using a logistic function. Interindividual differences in resting heart rates and blood pressures were observed, but all seven animals displayed clear baroreflex responses. Atropine and sotalol greatly attenuated the response. A maximal baroreflex gain of 7.2 beats min(-1) kPa(-1) was found at a mean aortic pressure of 6.1 kPa, indicating the active role of the baroreflex in a wide pressure range encompassing hypotensive and hypertensive states. At the lowest mean aortic pressures (5.0 kPa), the synergistic role of the pulmonary-to-systemic shunt in buffering the blood pressure drop also contributes to blood pressure regulation, Pulse pressure showed a better correlation,vith heart rate and also a higher gain than mean aortic, systolic or diastolic pressures, and this is taken as an indicator of the existence of a differential control element working simultaneously with a linear proportional element.

Hypoxia stimulates cerebral blood flow in the estuarine crocodile (Crocodylus porosus)

The effect of N-2 respiration on cerebral blood flow (CBF) velocity on the dorsal surface of cerebellum was examined in the estuarine crocodile, Crocodylus porosus, using epi-illumination microscopy. Twelve minutes of N-2 respiration resulted in a 126% increase in CBF velocity. N-2 respiration had no effect on blood pressure, indicating an underlying cerebral vasodilation. In addition, heart rate increased significantly. Systemic injections of aminophylline and the NO synthase (NOS) inhibitor nitro-L-arginine (L-NA) did not affect the hypoxia induced increase in CBF. We conclude that C. porosus responds to hypoxia with adenosine and nitric oxide (NO) independent cerebral vasodilation, and that this is likely to be a mechanism protecting the brain from energy deficiency during prolonged dives. (C) 1999 Elsevier Science Ireland Ltd. All rights reserved.

From anatomy to angioscopy: 164 years of crocodilian cardiovascular research, recent advances, and speculations

This paper present: a synthesis of current research on the crocodilian cardiovascular system with a view to encourage discussion and debate about the intricacies of this unique system and to provide ideas and suggestions for future studies. Innovative experimental approaches combined with new technologies have helped to resolve the complex flow and pressure patterns observed during non-shunting conditions that predominate in resting instrumented animals and during pulmonary to systemic shunting, which has been observed to occur spontaneously and during diving. The mechanisms and structures that may induce and regulate shunting are presented and the functional significance of a pulmonary to systemic shunt is discussed. (C) 1997 Elsevier Science Inc.

Notopronocephalus peekayi gen. et sp. n. (Digenea: Pronocephalidae) from Australian freshwater turtles

Notopronocephalus peekayi gen, et sp, n. is described from the intestine of Elseya latisternum Gray, 1867, E. dentata (Gray, 1863) and Emydura signata Ahl, 1932 from rivers in Queensland. The new genus is distinguished by the absence of ventral glands, simple (neither diverticulate nor sinuous) caeca terminating at the anterior margin of the testes, excretory arms not uniting in forebody, single ovary, two opposite testes close to the posterior end of the body, intracaecal genital pore, vitelline follicles anterior to the testes, cirrus-sac orientated obliquely and not divided into two portions, and the uterus intracaecal. This is the first pronocephalid to be described from an Australian freshwater turtle and the first from the family Chelidae.