Reptilian Blood Research Articles

Molecular Screening of Haemogregarine Hemoparasites (Apicomplexa: Adeleorina: Haemogregarinidae) in Populations of Native and Introduced Pond Turtles in Eastern Europe.

Haemogregarines (Apicomplexa: Adeleorina) are the most common and widespread reptilian blood parasites. Haemogregarina stepanowi was the first haemogregarine described from a reptile, the European pond turtle Emys orbicularis, and initial assessments indicated it was widespread across different pond turtle host species across much of Europe, the Middle East and North Africa. However, recent molecular assessments have indicated the presence of multiple genetically distinct forms in North Africa and the Iberian Peninsula, and extensive mixed infections which may be associated with a negative impact on the hosts. Here, we screened two native species, E. orbicularis and Mauremys rivulata, and the introduced Trachemys scripta from Serbia and North Macedonia for haemogregarines by amplifying and sequencing part of the 18S rRNA gene of these parasites, and used a standard DNA barcoding approach to identify leeches, the final host, attached to pond turtles. Our results again demonstrate the occurrence of considerable diversity of parasites in the analysed pond turtle species, and that T. scripta are likely infected by local haemogregarine parasites, and not those that are found in its native range. Leeches were identified as Placobdella costata, part of a lineage from Northern Europe. Mixed infections within pond turtles were again common. Current haemogregarine taxonomy does not reflect the genetic diversity identified, and a full taxonomic reassessment is needed.

Experimental feeding of Sergentomyiaminuta on reptiles and mammals: comparison with Phlebotomuspapatasi

BackgroundSergentomyiaminuta (Diptera: Phlebotominae) is an abundant sand fly species in the Mediterranean basin and a proven vector of reptile parasite Leishmania (Sauroleishmania) tarentolae. Although it feeds preferentially on reptiles, blood meal analyses and detection of Leishmania (Leishmania) infantum DNA in wild-caught S.minuta suggest that occasional feeding may occur on mammals, including humans. Therefore, it is currently suspected as a potential vector of human pathogens.MethodsA recently established S.minuta colony was allowed to feed on three reptile species (i.e. lizard Podarcissiculus and geckos Tarentolamauritanica and Hemidactylusturcicus) and three mammal species (i.e. mouse, rabbit and human). Sand fly mortality and fecundity were studied in blood-fed females, and the results were compared with Phlebotomuspapatasi, vector of Leishmania (L.) major. Blood meal volumes were measured by haemoglobinometry.ResultsSergentomyiaminuta fed readily on three reptile species tested, neglected the mouse and the rabbit but took a blood meal on human. However, the percentage of females engorged on human volunteer was low in cage (3%) and feeding on human blood resulted in extended defecation times, higher post-feeding mortality and lower fecundity. The average volumes of blood ingested by females fed on human and gecko were 0.97 µl and 1.02 µl, respectively. Phlebotomuspapatasi females readily fed on mouse, rabbit and human volunteer; a lower percentage of females (23%) took blood meal on the T.mauritanica gecko; reptilian blood increased mortality post-feeding but did not affect P.papatasi fecundity.ConclusionsAnthropophilic behaviour of S.minuta was experimentally demonstrated; although sand fly females prefer reptiles as hosts, they were attracted to the human volunteer and took a relatively high volume of blood. Their feeding times were longer than in sand fly species regularly feeding on mammals and their physiological parameters suggest that S.minuta is not adapted well for digestion of mammalian blood. Nevertheless, the ability to bite humans highlights the necessity of further studies on S.minuta vector competence to elucidate its potential role in circulation of Leishmania and phleboviruses pathogenic to humans.Graphical abstract

HEMATOLOGIC AND BIOCHEMICAL VALUES OF FREE-RANGING HAWKSBILL SEA TURTLES (ERETMOCHELYS IMBRICATA) IN GLOVER'S REEF, BELIZE.

Blood samples were obtained from the dorsal cervical sinus of free-ranging hawksbill sea turtles (Eretmochelys imbricata) collected at Glover's Reef Marine Reserve, 42 km east of the coast of Belize, for hematology and plasma biochemistry analysis. Unknown sex, subadult turtles (N = 32) were sampled in 2013 (n = 22) and 2017 (n = 10). To provide a more robust data set, parameters that did not have statistically significant differences were pooled and treated as a single population. Eleven hematologic parameters were evaluated; of these, five were pooled. Twenty-three plasma biochemical parameters were evaluated; of these, 15 were pooled. The PCV observed in this study (mean 33.44%) was double that observed in two studies of juvenile hawksbills in Dubai (means 17% and 16%), whereas the total WBC count was half that observed in immature and adult hawksbills sampled in the Galápagos (mean 2.91 × 103 versus 5.3 × 103/µl). Total protein and albumin were lower than regionally similar, adult female hawksbills in Brazil (means 3.36 versus 5.45 g/dl and 0.93 versus 2.11 g/dl, respectively). Globulins were higher (mean 2.43 versus 1.06 and 0.5 g/dl), driving the albumin:globulin ratio lower than that observed in two studies of juvenile hawksbills in Dubai (0.4:1 versus 1.1:1 and 1:1, respectively). These findings represent a geographically distinct population from previous reports, highlight the variability in blood parameters from disparate populations, and reaffirm the vital importance of considering a multitude of variables when interpreting reptilian blood values. The similarities in the majority of values observed in 2013 and 2017 provide confidence in the stability of these parameters in this population.

Leishmania (Sauroleishmania) tarentolae isolation and sympatric occurrence with Leishmania (Leishmania) infantum in geckoes, dogs and sand flies

The trypanosomatid protist Leishmania tarentolae is a saurian-associated parasite vectored by the Sergentomyia minuta sand fly. This study aimed to confirm the circulation of L. infantum and L. tarentolae in sand flies, reptiles and dogs and to isolate new strains of these protists. Reptilian and sheltered dog blood samples were collected, and sand flies were captured. Samples were tested for Leishmania spp. using duplex real-time PCR (dqPCR) and real-time PCR (qPCR); the origin of blood meal was identified in engorged sand flies by conventional PCR. The reptilian blood and intestinal content of sand fly females were cultured. Dog sera were tested by IFAT using both Leishmania species. Four Tarentola mauritanica geckoes were molecularly positive for L. infantum or L. tarentolae, with no co-infections; moreover, amastigote-like forms of L. infantum were observed in the bone marrow. 24/294 sand flies scored positive for Leishmania spp. by dqPCR, 21 S. minuta and two Phlebotomus perniciosus were positive for L. tarentolae, while only a single Ph. perniciosus was positive for L. infantum. Blood meal analysis confirmed reptile and dog in S. minuta, dog and human in Ph. perniciosus and dog in Phlebotomus neglectus. Two axenic strains of L. tarentolae were obtained. Twelve of 19 dogs scored positive for L. infantum and L. tarentolae by IFAT and three of them also for L. infantum by dqPCR, and six by qPCR. These data confirm the sympatric circulation of L. infantum and L. tarentolae in geckoes, sand flies, and dogs, and suggest that geckoes may be infected with L. infantum.

A simple CO2 equilibration method for measuring blood oxygen isotope compositions.

Blood water oxygen isotope compositions can provide valuable insights into physiological processes and ecological patterns. While blood samples are commonly drawn for medical or scientific purposes, blood fractions are infrequently measured for oxygen isotopic compositions (δ18 O) because such measurements are time consuming and expensive. We sampled blood from sheep, goats, and iguanas raised in field and animal laboratories into serum, EDTA, heparin, and uncoated plastic vials commonly used in medical and scientific research, then separated red blood cell (RBC) and plasma or serum blood fractions. These were injected into helium-flushed Exetainer tubes where they naturally outgassed endogenous CO2 (goat blood), or into He- and CO2 -flushed tubes (iguana blood). The CO2 gas was sampled on a GasBench II system, and δ18 O was measured by an isotope ratio mass spectrometer (IRMS). Repeated δ18 O measurements were stable over multiple days. The addition of desiccated blood solids to water standards had little impact on their δ18 O measurements, suggesting that organic molecular constituents within blood serum and plasma do not interfere with blood water δ18 O values. We observed slight but statistically significant δ18 O offsets between plasma, serum and RBC fractions. Mass-dependent body water turnover times for iguanas were derived from the data. We demonstrate that a simple blood-CO2 equilibration method using the GasBench can quickly, reliably and accurately characterize water δ18 O in the plasma, RBC, and whole blood fractions of mammalian and reptilian blood samples (precision ≤ 0.1‰). This method will expand the application of blood stable isotope analysis in physiological and medical research.

Identification of trypanosomatids and blood feeding preferences of phlebotomine sand fly species common in Sicily, Southern Italy.

In this study, the presence of Leishmania DNA and blood feeding sources in phlebotomine sand fly species commonly present in Sicily were investigated. A total of 1,866 female sand flies including 176 blood fed specimens were sampled over two seasons in five selected sites in Sicily (southern Italy). Sergentomyia minuta (n = 1,264) and Phlebotomus perniciousus (n = 594) were the most abundant species at all the sites, while three other species from the genus Phlebotomus (i.e., P. sergenti n = 4, P. perfiliewi n = 3 and P. neglectus n = 1) were only sporadically captured. Twenty-eight out of the 1,866 (1.5%) sand flies tested positive for Leishmania spp. Leishmania tarentolae DNA was identified in 26 specimens of S. minuta, while the DNA of Leishmania donovani complex was detected in a single specimen each of S. minuta and P. perniciosus. Interestingly, seven S. minuta specimens (0.4%) tested positive for reptilian Trypanosoma sp. Blood sources were successfully identified in 108 out of 176 blood fed females. Twenty-seven out of 82 blood sources identified in fed females of P. perniciosus were represented by blood of wild rabbit, S. minuta mainly fed on humans (16/25), while the sole P. sergenti fed specimen took a blood meal on rat. Other vertebrate hosts including horse, goat, pig, dog, chicken, cow, cat and donkey were recognized as blood sources for P. perniciosus and S. minuta, and, surprisingly, no reptilian blood was identified in blood-fed S. minuta specimens. Results of this study agree with the well-known role of P. perniciosus as vector of L. infantum in the western Mediterranean; also, vector feeding preferences herein described support the hypothesis on the involvement of lagomorphs as sylvatic reservoirs of Leishmania. The detection of L. donovani complex in S. minuta, together with the anthropophilic feeding-behaviour herein observed, warrants further research to clarify the capacity of this species in the transmission of pathogens to humans and other animals.

Clinical Management of an Intranasal Abscess in a Captive Elongate Tortoise (Indotestudo elongata) by Using Modified Choukroun's Platelet-Rich Fibrin

An adult male elongate tortoise (Indotestudo elongata) was presented to the Animal Medical Center of Taipei Zoo with an abscess on the tip of its nose. After 43 days of daily debridement and systemic antibiotic administration, we sought an alternative treatment modality due to poor wound management. We developed a novel treatment using platelet-rich fibrin (PRF). There are few studies documenting the benefits of PRF for the management of wounds in reptiles. Most published protocols require the use of specific anticoagulants and methods to prepare PRF; however, we found that a modified Choukroun's PRF was relatively easy to prepare and use for wound treatment of an elongate tortoise. Here, we describe a successful outcome of a chronic, nonresolving facial wound after treatment with modified Choukroun's PRF. To our knowledge, this is the first report of a Choukroun's PRF preparation derived from reptilian blood. We recommend further studies and detailed trials to evaluate the efficacy of this protocol in reptiles.

A Step-by-Step Guide to Assemble a Reptilian Genome.

Multiple technologies and software are now available facilitating the de novo sequencing and assembly of any vertebrate genome. Yet the quality of most available sequenced genomes is substantially poorer than that of the golden standard in the field: the human genome. Here, we present a step-by-step protocol for the successful sequencing and assembly of a high-quality snake genome that can be applied to any other reptilian or avian species. We combine the great sequencing depth and accuracy of short reads with the use of different insert size libraries for extended scaffolding followed by optical mapping. We show that this procedure improved the corn snake scaffold N50 from 3.7kbp to 1.4Mbp, currently making it one of the snake genomes with the longest scaffolds. Short guidelines are also given on the extraction of long DNA molecules from reptilian blood and the necessary modifications in DNA extraction protocols. This chapter is accompanied by a website ( www.reptilomics.org/stepbystep.html ), where we provide links to the suggested software, examples of input and output files, and running parameters.

Flow cytometric characterization of peripheral blood leukocyte populations of 3 neotropical snake species: Boa constrictor, Bothrops jararaca, and Crotalus durissus.

The reptilian immune system is represented by innate, humoral, and cell-mediated mechanisms, involving different types of blood leukocytes. The development of optimized methods for the advanced study of origin and function of reptilian blood leukocytes is needed. The purpose of the study was to optimize leukocyte density gradient isolation protocols from snake peripheral blood samples, and characterize recovered cells by flow cytometry based on size and internal complexity for a qualitative and semi-quantitative assessment of leukocyte populations in one boa (Boa constrictor), and 2 viper species (Bothrops jararaca, Crotalus durissus). Blood samples from 30 snakes (10 from each species, 5 males and 5 females) were collected in tubes with sodium heparin. Fresh blood was centrifuged with either ficoll-paque PLUS or percoll density gradients for leukocyte isolation. Flow cytometric leukocyte gates were defined based on size (forward scatter [FSC]) and internal complexity (side scatter [SSC]). Relative leukocyte differential counts after sorting the cells in these gates in one snake for each species were compared to conventional light microscopic differential counts on unsorted isolated leukocytes. There was no statistical difference in the relative leukocyte populations, including heterophils, azurophils, and small and large lymphocytes between samples isolated by ficoll or percoll. Four leukocyte gates were identified based on their location in FSC/SSC cytograms. The relative leukocyte differential counts after sorting in single animals showed some agreement with the light microscopy differential count on unsorted cells. Based on FSC and SSC, 4 distinct leukocyte populations were found in ficoll or percoll density gradient isolated leukocytes from peripheral blood from boa and viper species. Further optimization of the technique should allow the performance of functional assays.

Summary of oxygen transport characteristics of reptilian blood

Summary of oxygen transport characteristics of reptilian blood

Anticoagulants for avian and reptilian blood: heparin and EDTA.

The effectiveness of EDTA (dipotassium and disodium salts) and heparin as anticoagulants for the blood of puffadders and pigeons has been investigated. In the case of the snakes all three substances produced haemolysis after variable time intervals, and increases in cell volume were observed. In the case of the pigeons, samples stored at 4 degrees C showed lower haematocrit values and haemolysed sooner than samples stored at room temperature.

Leucocyte and Erythrocyte Diluent for Reptilian Blood Cell Counts

Leucocyte and Erythrocyte Diluent for Reptilian Blood Cell Counts

Haemogregarines in Snakes: The Incidence and Identity of the Erythrocytic Stages

The traditional methods of displaying reptile specimens in zoos provide ideal conditions for transmission of protozoan parasites due to the humid cage atmosphere and the sand or leaf mulch substrates used to simulate natu.ral habitats. These physical conditions favor arthropod reproduction; the concentration of potential hosts favors transmission of whatever parasites may be present. Indeed, the mortality rates typical of zoo reptile collections substantiate the probability that more than just the nutritional and psychological effects of confinement and display affect the longevity of the captive population. Only impractically elaborate precautions could eliminate the possibility of parasite transmission under the display conditions deemed necessary to assuage the aesthetic sensibilities of the general public. Therefore, examination of zoo specimens for parasites may be expected to be quite rewarding; seldom are conditions contributing to high parasite concentrations combined with the ready availability of hosts from all over the world that a zoo offers. This report presents data concerning the incidence and identity in captive snakes of the protozoan blood parasite generally included in the genus Haemnogregarina. These data were gathered during 1955 through 1958 as part of a program concerning experimental transmission of reptilian blood parasites. The authors are grateful to the curators of the reptile collections at the Lincoln Park Zoo, Chicago, Illinois, the Toledo Zoo, Toledo, Ohio, the Cleveland Municipal Zoo, Cleveland, Ohio, and to the zoo personnel at these institutions for their physical aid and moral support in the pursuit of the specimens from which blood and these data are taken.

Some physiological properties of reptilian blood

Some physiological properties of reptilian blood

Properties of reptilian blood. IV. The alligator (alligator mississippiensis daudin)

Journal of Cellular and Comparative PhysiologyVolume 6, Issue 2 p. 243-254 Article Properties of reptilian blood. IV. The alligator (alligator mississippiensis daudin) D. B. Dill, D. B. Dill Fatigue Laboratory, Morgan Hall, Harvard University, BostonSearch for more papers by this authorH. T. Edwards, H. T. Edwards Fatigue Laboratory, Morgan Hall, Harvard University, BostonSearch for more papers by this author D. B. Dill, D. B. Dill Fatigue Laboratory, Morgan Hall, Harvard University, BostonSearch for more papers by this authorH. T. Edwards, H. T. Edwards Fatigue Laboratory, Morgan Hall, Harvard University, BostonSearch for more papers by this author First published: June 1935 https://doi.org/10.1002/jcp.1030060206Citations: 29AboutPDF ToolsRequest permissionExport citationAdd to favoritesTrack citation ShareShare Give accessShare full text accessShare full-text accessPlease review our Terms and Conditions of Use and check box below to share full-text version of article.I have read and accept the Wiley Online Library Terms and Conditions of UseShareable LinkUse the link below to share a full-text version of this article with your friends and colleagues. Learn more.Copy URL Share a linkShare onFacebookTwitterLinkedInRedditWechat Citing Literature Volume6, Issue2June 1935Pages 243-254 RelatedInformation

Properties of reptilian blood. III. The chuckwalla (sauromalus obesus baird)

Journal of Cellular and Comparative PhysiologyVolume 6, Issue 1 p. 37-42 Article Properties of reptilian blood. III. The chuckwalla (sauromalus obesus baird) D. B. Dill, D. B. Dill Fatique Laboratory, Morgan Hall, Harvard University, BostonSearch for more papers by this authorH. T. Edwards, H. T. Edwards Fatique Laboratory, Morgan Hall, Harvard University, BostonSearch for more papers by this authorA. V. Bock, A. V. Bock Fatique Laboratory, Morgan Hall, Harvard University, BostonSearch for more papers by this authorJ. H. Talbott, J. H. Talbott Fatique Laboratory, Morgan Hall, Harvard University, BostonSearch for more papers by this author D. B. Dill, D. B. Dill Fatique Laboratory, Morgan Hall, Harvard University, BostonSearch for more papers by this authorH. T. Edwards, H. T. Edwards Fatique Laboratory, Morgan Hall, Harvard University, BostonSearch for more papers by this authorA. V. Bock, A. V. Bock Fatique Laboratory, Morgan Hall, Harvard University, BostonSearch for more papers by this authorJ. H. Talbott, J. H. Talbott Fatique Laboratory, Morgan Hall, Harvard University, BostonSearch for more papers by this author First published: April 1935 https://doi.org/10.1002/jcp.1030060104Citations: 20AboutPDF ToolsRequest permissionExport citationAdd to favoritesTrack citation ShareShare Give accessShare full text accessShare full-text accessPlease review our Terms and Conditions of Use and check box below to share full-text version of article.I have read and accept the Wiley Online Library Terms and Conditions of UseShareable LinkUse the link below to share a full-text version of this article with your friends and colleagues. Learn more.Copy URL Share a linkShare onFacebookTwitterLinked InRedditWechat Citing Literature Volume6, Issue1April 1935Pages 37-42 RelatedInformation

Properties of reptilian blood. II. The gila monster (heloderma suspectum cope)

Journal of Cellular and Comparative PhysiologyVolume 6, Issue 1 p. 21-35 Article Properties of reptilian blood. II. The gila monster (heloderma suspectum cope) H. T. Edwards, H. T. Edwards Fatigue Laboratory, Morgan Hall, Harvard University, BostonSearch for more papers by this authorD. B. Dill, D. B. Dill Fatigue Laboratory, Morgan Hall, Harvard University, BostonSearch for more papers by this author H. T. Edwards, H. T. Edwards Fatigue Laboratory, Morgan Hall, Harvard University, BostonSearch for more papers by this authorD. B. Dill, D. B. Dill Fatigue Laboratory, Morgan Hall, Harvard University, BostonSearch for more papers by this author First published: April 1935 https://doi.org/10.1002/jcp.1030060103Citations: 21AboutPDF ToolsRequest permissionExport citationAdd to favoritesTrack citation ShareShare Give accessShare full text accessShare full-text accessPlease review our Terms and Conditions of Use and check box below to share full-text version of article.I have read and accept the Wiley Online Library Terms and Conditions of UseShareable LinkUse the link below to share a full-text version of this article with your friends and colleagues. Learn more.Copy URL Citing Literature Volume6, Issue1April 1935Pages 21-35 RelatedInformation