Marcos Tavares-Dias

Biochemical and hematological reference ranges for Amazon freshwater turtle, Podocnemis expansa (Reptilia: Pelomedusidae), with morphologic assessment of blood cells.

Blood parameters are useful to measure physiological disturbances in chelonians. Thus they can provide important information for the diagnosis and prognosis of diseases. The aim of this paper was to determine the reference range of plasma glucose, total protein, triglycerides, cholesterol and urea levels, as well as hematocrit, hemoglobin, red blood cell, thrombocyte and white blood cell counts for healthy Podocnemis expansa (Schweigger, 1812), bred in a turtle farm in the Amazonas State, Brazil. Plasma glucose, hemoglobin, mean corpuscular volume (MCV), lymphocytes and heterophils were the parameters with the smallest variations. Significant (p<0.001) correlations between red blood cells count and hematocrit and hemoglobin concentration were found. Turtle P. expansa had lymphocytes, azurophils, heterophils, eosinophils and basophils with morphologic features similar to the ones of others turtles species. No investigated blood parameter was influenced by sex. In further studies, the established reference ranges might be useful for the health assessment of this turtle species.

Characterization of the acute inflammatory response in the hybrid tambacu (Piaractus mesopotamicus male × Colossoma macropomum female) (Osteichthyes)

This work evaluated the acute inflammatory response induced by injections of 0.5 mL saline solution (control), 500 microg carrageenin and 0.5 mL thioglycollate 3% in the swim bladder of juvenile tambacu hybrid. Fish were distributed in three treatments, three replications and acclimated for a period of 10 days before assay. The cell characterization from the inflammatory exudate was performed in Giemsa and PAS stained smears. Carrageenin, injected in fish, showed an increase on the total number of cells in the inflammatory exudate when compared to saline and thioglycollate injected. Whereas, for carrageenin-injected fish, the percentage of thrombocyte was higher than thioglycollate. On the other hand, granulocyte percentage in thioglycollate-injected fish was higher than the ones injected using carrageenin. Carrageenin provoked the highest migration of macrophage to the inflammatory site. The PAS method confirmed the presence of three types of granulocytes: eosinophilic granular cell (EGC) type 1 with the characteristics of a special granulocytic cell commonly found in the circulating blood; EGC type 2 shorter than the last one and neutrophil. This study contributes to a better understanding of the inflammatory response and infectious processes in native fish.

Methodological limitations of counting total leukocytes and thrombocytes in reptiles (Amazon turtle, Podocnemis expansa): an analysis and discussion

The aim of this paper is to compare three different methods for counting white blood cells [WBC] (Natt and Herrick method, estimation with 1,000 and 2,000 erythrocytes) and three methods for counting total thrombocytes [TT] (Wojtaszek method, estimation with 1,000 and 2,000 erythrocytes) in a South American freshwater turtle species, Podocnemis expansa, Schweigger 1812 (Reptilia, Pelomedusidae). Direct WBC counts using the Natt and Herrick method showed limitations, which are discussed here. The WBC and TT counts using 1,000 erythrocytes from blood smears are not recommended for Amazon turtles nor other reptilian species, since wide variation in counts can be observed. Estimation methods for determining WBC and TT based on 2,000 erythrocytes of blood smears were most acceptable because they allow a differentiation between leukocytes and thrombocytes and also had a smaller variation. The methods investigated here for the Amazon turtle, which have been widely used in other reptile species, provided evidence that the most acceptable method is not that of using diluted stains and a hemocytometer.

Fluorescence Detection of the Ornamental Fish Cardinal Tetra (Paracheirodon axelrodi)

The fluorescence spectra of the tropical fish, Cardinal Tetra (Paracheirodon axelrodi), originating in the Amazon region of Brazil, were determined. These spectra were then treated using factor analysis, generating two contributing spectra and separating out the noise. Time‐resolved fluorescence results indicated that the fluorescent system in the epidermis undergoes excited state reaction. Excited state proton transfer is suggested as being present. Both intentionally stressed and nonstressed individuals were used and some small differences were noted in the contributions of the two calculated contributing spectra to the experimental spectra, presumably as a function of stress. The results are compared with those obtained by the standard determination of cortisol level using the whole body extraction method and it is suggested that the method could be tested as an improved, nondestructive way to determine stress in this species, which is a necessary step in the development of “best management practices” of methods for storage and transport of the fish.

Ecophysiological interactions and water-related physicochemical parameters among freshwater stingrays

The objective of this study was to compare and correlate the ecology of neonates and young individuals of Potamotrygon wallacei, Potamotrygon motoro and Paratrygon aiereba with regard to their hematological profile and the physicochemical parameters of the water that they inhabit. Principal component analysis (PCA) on the complete blood count revealed total variation of 72.92%, thus demonstrating a differentiation system for oxygen demand. On the other hand, P. motoro was considered to be an intermediate species, given that its complete blood count characteristics interacted with both P. wallacei and with P. aiereba. The interaction among the biochemical variables was shown to total 64.67% of the factors. This allowed differentiation of P. wallacei from P. aiereba, while P. motoro maintained an intermediate position. These characteristics of differentiation within the preferred environment corroborate the PCA of the present study and confirm that these species can be differentiated through considering the complete blood count and biochemical parameters. The PCA on water properties showed 68.57% differentiation, mainly comprising the x axis (49.44%). It can be affirmed that P. motoro has the capacity to inhabit the preferential areas of P. wallacei and P. aiereba, as well as occupying localities in which other stingrays are not found. In conclusion, P. wallacei presents patterns differentiating it from P. aiereba, while P. motoro is a species that presents intermediate characteristics. The latter can be considered to be a more broadly distributed species regarding its ecophysiological characteristics.

Cyrilia sp. (Apicomplexa: Haemogregarinidae) in the Amazonian freshwater stingray Potamotrygon wallacei (cururu stingray) in different hydrological phases of the Rio Negro

Intraerythrocytic parasites are frequently found in fish, including elasmobranchs. The Amazonian rivers present well defined annual hydrological cycles that results in drastic modifications of the environmental conditions with deep implications in the life cycle of the whole associated biota in those fluvial systems. The freshwater stingray Potamotrygon wallacei (stingray cururu) is a new species restricted to the Middle Rio Negro basin and it is subject to strong alterations in their natural habitats (igapós) a result of the constant variations in the water level of Rio Negro. This work demonstrates the occurrence of intraerythrocytic parasite Cyrilia sp. in this stingray species. Additionally, the prevalence and quantification of hemoparasites in different phases of Rio Negro were also established. Field sampling was carried in the Archipelago of Mariuá, Middle Rio Negro, involving different stages of the water cycle. The intraerythrocytic parasites were quantified by direct counting in blood smears using a total counting of 2000 erythrocytes in each blood smear. The presence of parasites intraerythrocytic generates changes in the morphology of blood cell. The largest amount of the hemoparasites was recorded in the drought period. We observed a decreasing tendency in the number of parasites in the blood between the drought periods and inundation. We concluded that the level of Negro River influences the incidence of intraerythrocytic parasites in the cururu stingray and the drought represents the period of larger susceptibility to the infestation.