Morphological, cytochemical, and ultrastructural observations on the blood cells of the reptile Tupinambis merianae (Squamata)

Tupinambis merianae is a lizard of the Brazilian fauna and belongs to the Squamata order. Blood cell data are scarce. Blood samples from six specimens of adult T. merianae were used to evaluate some hematological parameters. For structural analysis, 2 ml of blood was collected in the presence of EDTA. Part of the blood was used for preparing blood smears which were submitted to the methods of Leishman, Laür, and toluidine blue, and the cytochemical reactions of periodic acid-Schiff, sirius red, sudan black B, and ortho-toluidine-H₂O₂. The remainder was centrifuged and the leukocyte buffy coat was fixed in Karnovsky’s fluid for electron microscopy examination. The following blood cells were identified: mature and immature erythrocytes, spherical and elliptical thrombocytes, heterophilic granulocytes, eosinophils and basophils types I and II, lymphocytes, monocytes, and azurophilic cells. The more significant results obtained were: the presence of glycogen in the cytoplasm of the thrombocytes, heterophils, and basophils; the presence of basic polyaminoacid-rich proteins in the granules of heterophils and eosinophils and myeloperoxidase in the granules of the heterophils, eosinophils, and azurophilic cells; and sudanophilic small granules in the heterophils, eosinophils, and azurophilic cells. More detailed morphological aspects of the cells were observed by means of ultrastructural analysis.     

Haematology and serum biochemistry of golden eagle (Aquila chrysaetos) in Iran

Haematological and serum biochemical values were estimated in blood samples collected from 21 apparently adult golden eagles (Aquila chrysaetos) of both sexes. The mean values of red blood cells, packed cell volume, haemoglobin, white blood cells, heterophils, lymphocytes, monocytes and eosinophils were 1.63 ± 0.11 × 10¹²/l, 0.47 ± 0.009 l/l, 91.73 ± 1.52 g/l, 24.31 ± 1.97 × 10⁹/l, 4.40 ± 0.22 × 10⁹/l, 16.81 ± 0.65 × 10⁹/l, 0.99 ± 0.19 × 10⁹/l and 2.10 ± 0.30 × 10⁹/l, respectively. The leucocytes had 69.14%, 4.09%, 18.12% and 8.65% lymphocytes, monocytes, heterophils and eosinophils, respectively. The results of serum biochemistry in the golden eagle indicated that the concentrations of glucose, total protein, albumin, total globulin, cholesterol, triglyceride, uric acid, blood urea nitrogen, creatinine, calcium, phosphorous, aspartate aminotransferase, alanine aminotransferase, creatine kinase, lactate dehydrogenase and alkaline phosphatase were 16.42 ± 0.73 mmol/l, 49.76 ± 1.35 g/l, 20.46 ± 0.79 g/l, 29.30 ± 1.47 g/l, 2.14 ± 0.09 mmol/l, 2.04 ± 0.08 mmol/l, 457.67 ± 97.46 μmol/l, 2.74 ± 0.17 mmol/l, 53.27 ± 3.87 μmol/l, 2.37 ± 0.24 mmol/l, 1.73 ± 0.08 mmol/l, 293.24 ± 18.96 IU/l, 28.21 ± 2.36 IU/l, 411.29 ± 58.37 IU/l, 1,209.89 ± 21.73 IU/l and 67.31 ± 5.29 IU/l, respectively. There were no significant differences between haematological and serum biochemical parameters of male and female golden eagles (P > 0.05).     

Haematological and blood chemical values from Bothrops ammodytoides (ophidia-crotalidae) in captivity

In order to establish reference haematological and blood chemistry parameters, blood samples were obtained from 50 healthy specimens ofBothrops ammodytoides kept in captivity. The haematological parameters determined were: red blood cell count (RBC); total leucocyte (WBC) and differential leucocyte cell count; thrombocyte count; haematocrit (PCV); haemoglobin concentration; mean corpuscular volume (MCV); mean corpuscular haemoglobin (MCH) and mean corpuscular haemoglobin concentration (MCHC). Blood chemistry parameters measured were: total protein; albumin; globulins; glucose; urea; uric acid; triglycerides; cholesterol; calcium; phosphorus; magnesium; sodium; potassium and chloride concentrations and the activities of aspartate amino transferase (AST) and alanine amino transferase (ALT). Compared to the values published for other ophidian species, RBC count, PCV and WBC count inBothrops ammodytoides are lower than in most of the crotalids. Total protein and glucose concentrations are lower, whereas uric acid concentration, AST and ALT levels are higher than the values reported for other species.     

Haematology and serum biochemistry of the Carpentarian Rock-rat (<Emphasis Type="Italic">Zyzomys palatalis</Emphasis>)

Haematology and serum biochemistry was analysed on samples collected from four endangered Carpentarian Rock-rats (Zyzomys palatalis) maintained in a captive colony at the Territory Wildlife Park, Northern Territory. As these animals are endangered, only small sample numbers were able to be analysed. Most haematological and serum biochemical parameters were similar to those for other native Australian murids, with the exception of alkaline phosphatase and creatinine. The study provides preliminary haematology and serum biochemistry information for the Carpentarian Rock-rat and will assist managers of this species to assess the health of the captive and wild populations.     

Haematology of the fire-bellied toad,Bombina bombina L.

Haematological parameters and cytomorphometry of circulating blood were studied in the fire-bellied toad (Bombina bombina L.). The mean red blood cell count was significantly higher in males (0.340 × 10¹²/l) than in females (0.290 × 10¹²/l). Haemoglobin concentration was 7.11 g/dl, haematocrit 0.195 l/l, red blood cell indices MCV = 607 fl; MCH = 218.5 pg; MCHC = 367.8 g/l, thrombocyte count 6.275 × 10⁹/l, white blood cell count 8.382 × 10⁹/l, the composition being lymphocytes 4.34 × 10⁹/l; monocytes 0.97 × 10⁹/l; neutrophils 2.10 × 10⁹/l; basophils 0.90 × 10⁹/l; eosinophils 0.30 × 10⁹/l. Size, surface area and staining affinities of blood cells were also determined. Erythroblasts (7.44 × 10⁹/l) at various development stages in circulating blood of B. bombina constituted approximately 2.4% of the total number of mature red blood cells and were more numerous in males. The morphology of blood cells is presented as photomicrographs. Results of comparison with the haematology of the closest relatives of B. bombina are discussed.Abbreviations ERBC erythroblasts - Hb haemoglobin - HCT haematocrit - MCH mean cell haemoglobin - MCHC mean cell haemoglobin concentration - MCV mean cell volume - RBC red blood cell count - TC thrombocytes - WBC white blood cell count     

Normal haematology and blood chemistry of captive adult stone curlews ( Burhinus oedicnemus )

Blood samples were obtained from 18 stone curlews (Burhinus oedicnemus) as part of a study on health and disease in this species. Routine haematology and blood chemistry analyses were carried out to establish normal reference values for captive adult stone curlews. The results obtained are compared with information available for the Cape thick-knee (Burhinus capensis) and values originated from similar studies carried out in bustards.     

Haematology and red cell enzymes of the Australian orange-bellied parrot, Neophema chrysogaster

The haematology, red cell enzymes, and haemoglobin electrophoresis of twelve captive bred Australian orange-bellied parrots (Neophema chrysogaster) were studied. The means of the haematological parameters were: haemoglobin 15.3 g/dl, PCV 54%, red cell count 3.4 × 10¹²/l, MCV 138.0 fl, MCH 44.7 pg, leucocyte count 7.2 × 10⁹/l, heterophils 61%, lymphocytes 24%, monocytes 6.3%, eosinophils 0.7% and basophils 6.8%. The basophil count is higher than that recorded in other parrot species. The red cell enzyme activity is lower than those found in human red cells and similar to those recorded in the limited amount of data available for other avian species. Haemoglobin electrophoresis showed a typical avian pattern.     

Haematology, red cell enzymes, and red cell metabolic intermediates of 20 wild southern elephant seals (Mirounga leonina) from Macquarie Island

Blood was collected for haematological, red cell enzyme and red cell metabolic intermediate studies from 20 Southern elephant seals Mirounga leonina. Mean haematological values were: haemoglobin (Hb) 22.4 ± 1.4 g/dl, packed cell volume (PCV) 54.2 ± 3.8%, mean cell volume (MCV) 213.0 ± 5.0 fl and red cell count (RCC) 2.5 × 10¹²/l. Red cell morphology was unremarkable. Most of the red cell enzymes showed low activity in comparison with human red cells. Haemoglobin electrophoresis showed a typical pinniped pattern, i.e. two major components. Total leucocyte counts, platelet counts, and coagulation studies were within expected mammalian limits. Eosinophil counts varied from 0.5 × 10⁹/l to 7.7 × 10⁹/l(5%–49%), and there was a very wide variation in erythrocyte sedimentation rates, from 3 to 60 mm/h.     

An analysis of haematological findings on a feral population of aldabra giant tortoises (Geochelone gigantea)

In order to obtain haematological reference values for health assessment in captivity, blood samples were obtained from 143 feral Geochelone gigantea in a natural environment on Curieuse Island, Republic of the Seychelles. Tests were carried out by specially adapted field techniques. Because difficulty was experienced with obtaining a suitable blood sample, a method involving in vivo mixing was developed. Using this method it was found that values for haemoglobin (Hb), packed cell volume (PCV) and red (RBC) and white cell count (WBC) of healthy adult G. gigantea were low and the red cells were hypochromic and macrocytic compared with those of mammals and birds. Females had significantly lower Hb, RBC, PCV values and smaller red cells than males. Blood parasites were not found.     

Reticulo-endothelial stroma of the head-kidney from the seawater teleost gilthead seabream (Sparus aurata L.): An ultrastructural and cytochemical study

Background: Head-kidney, considered the major fish lympho-haemopoietic tissue, consists of cells of the different haemopoietic series supported by a network of stromal cells whose morphofunctional properties have not been established. We report the ultrastructure and cytochemical features of the reticulo-endothelial stroma of the head-kidney from the seawater teleost gilthead seabream (Sparus aurata L.). Methods: Samples of head-kidney were processed for electron microscopic study. Some of the samples were incubated for acid and alkaline phosphatase, peroxidase, glucose-6-phosphatase, or ATPase. Results: The reticulo-endothelial stroma of gilthead seabream head-kidney consists of sinusoidal cells (endothelial and adventitial cells) and reticular cells (macrophage-type reticulum and fibroblast-like reticular cells). Transcytosis vesicles and rounded medium electron-dense granules were observed in the cytoplasm of the endothelial cells. The adventitial cells partially covered the outside surface of the endothelial cells and were joined by desmosomes. The macrophage-type reticulum cells were characterized by their cytoplasmic processes and acid phosphatase positive lysosomes. The fibroblast-like reticular cells were joined by desmosomes and formed an extensive network between the haemopoietic parenchyma. They were peroxidase negative and acid and alkaline phosphatase, glucose-6-phosphatase, β-glucuronidase, and ATPase positive. Conclusions: The ultrastructural and cytochemical features of the reticulo-endothelial stroma of the gilthead seabream head-kidney are similar to those of mammalian bone marrow, suggesting phylogenetic analogies between both tissues. © 1995 Wiley-Liss, Inc.     

An ultrastructural study of the microtriches of adultProteocephalus tidswelli (Cestoda: Proteocephalidea)

Ultrastructural observations on adultProteocephalus tidswelli revealed a marked microtrichial polymorphism. Structural and dimensional variations of microtriches between different regions of the strobila and scolex, as well as within the same region, were observed. The authors suggest that microtriches are involved in a diversity of functions, and possible functional activities are discussed.Abbreviations A Apical organ - B Base of microthrix - BL Blade-like microthrix - BP Baseplate - C Cytoplasmic core - D Disc bodies - DC Distal cytoplasm - FL Filamentous microthrix - G Gravid segments - GBL Giant blade-like microthrix - GC Granular cytoplasm - GX Glycocalyx - IM Immature segments - M Matrix - MA Mature segments - MF Microfilaments - N Neck - PL Peg-like microthrix - PM Plasma membrane - R Rostellum - S Sucker - SH Shaft of microthrix - SL Spine-like microthrix - ST Strap-like structures - T Thick-walled tube     

Haematological, cytochemical and ultrastructural characteristics of blood cells and serum chemistry in captive Malayan tapir (Tapirus indicus) in Thailand

The Malayan tapir (Tapirus indicus) is classified as a preserved animal of Thailand. It is listed as an endangered species on the 2008 IUCN red list and also by the Convention on International Trade in Endangered Species of Wild Fauna and Flora. Blood samples from captive Malayan tapirs (n?=?20, 10 males and 10 females) from five zoos under the Zoological Park Organization of Thailand were examined and measured for complete blood counts, serum chemistry and electrolyte analysis. Cytochemical staining including Peroxidase, Sudan Black B, Periodic acid Schiff's reaction (PAS), α-naphthyl acetate esterase (ANAE) and β-glucuronidase (BG) was also analysed. Statistically significant differences were found between sexes including alkaline phosphatase, total protein and red cell distribution widths. Red blood cells were slightly anisocytosis, (range 4.5–8.5 μm) with 6.3-μm mean diameter. Eosinophils and monocytes were positive with all five cytochemical stains. Lymphocytes showed a variable reaction for ANAE and BG. PAS was positive for all type of blood cells except red blood cells. Ultrastructurally, eosinophils had the largest round shape with three-capsule layer granules which were prominent and distinguishable from other species. This investigation provides basic haematology and serum chemistry analyses as well blood cells identification guideline for the Malayan tapir and serves as endangered mammal reference values.     

The Complete CDS of the Prion Protein (<Emphasis Type="Italic">PRNP</Emphasis>) Gene of African Lion (<Emphasis Type="Italic">Panthera leo</Emphasis>)

We provide the complete PRNP CDS sequence for the African lion, which is different from the previously published sequence and more similar to other carnivore sequences. The newly obtained prion protein sequence differs from the domestic cat sequence at three amino acid positions and contains only four octapeptide repeats. We recommend that this sequence be used as the reference sequence for future studies of the PRNP gene for this species.     

Ultrastructural study of the endocrine cells of the gut of Testudo graeca (Chelonia)

Summary The digestive tract of Testudo graeca (Chelonia) was investigated by means of electron microscopy using both conventional and immunocytochemical techniques. EC-, L-, D-, G-, B-, N- and EC-L-cells were detected. These cells share several common ultrastructural characteristics with the endocrine cells of mammals (i.e. clear cytoplasm, prominent Golgi apparatus, secretory granules etc.). EC and D₁ cells have so far not been described in the esophagus of any animal species; in the present study these cells have been observed in the esophagus of T. graeca. Of special interest was the presence of B-cells in the intestine, suggesting that the migration of B-cells from the gut to the pancreas to constitute pancreatic islets is not concluded in T. graeca. The present study demonstrates that the gut endocrine system of T. graeca is a complex structure containing a large variety of endocrine cell types similar in morphology to those found in higher vertebrates.     

A convenient method for in situ processing of cultured cells for cytochemical localization by electron microscopy

The effects of various exogenous agents on subcellular structures is of importance to many investigators and can be critically evaluated by the use of cytochemical techniques and transmission electron microscopy. Therefore various cell culture techniques have become increasingly important in biological research in order to help determine these effects. Presently, most existing methods for processing anchorage-dependent cultures for electron microscopy utilize cells grown on either glass or plastic substrates. Therefore various coating substances have been applied to the culture surfaces to facilitate removal of the sample, however, incomplete separation and sample fracture often results. Here we present a simple method of in situ processing of samples for electron microscopy involving the use of detachable chamber slides. This method allows for the use of a quick processing procedure that results in a complete separation of the sample from the glass slide.     

Biological and mechanical quality of red blood cells cultured from human umbilical cord blood stem cells

Human umbilical cord blood (CB) has moved from the status of biological waste to that of a valuable source of haematopoietic stem (HS) cells. There are potentially three major clinical applications for HS cells andex vivo-expanded HS cells: reconstitution of haematopoiesis in patients undergoing chemotherapy; gene therapy (e.g. in thalassaemia, sickle cell anaemia); and large-scale production of mature blood cells. Erythropoiesis is accomplished by highly complex interactions of haematopoietic progenitor cells, stromal cells and cytokines in the bone marrow. Among them, erythropoietin is the principal regulator.Ex vivo cell culture experiments to obtain mature red blood cells were the focus of this study. Attempts to elucidate appropriate medium components and amounts of haematopoietic growth factors were successful: enucleated and haemoglobin-filled erythroid cells were obtained from primitive HS cells. Dimethylsulphoxide (DMSO) was found to be of particular importance as an efficient differentiation inducer. The differentiation process was followed microscopically and by fluorescence-activated cell sorting (FACS). Using the micropipette aspiration technique, the elastic properties of erythroid cells were evaluated as erythropoiesis progressed. Discocyte-like cells, comprising reticulocytes and finally differentiated red blood cells, showed an about ten-fold higher membrane shear modulus compared with control cells.     

An electron microscopic and cytochemical study ofLeptomonas samueli

The fine structure of promastigotes ofLeptomonas samueli is described. This protozoan revealed several features in common with other trypanosomatids. A large membrane-bound cavity containing many vesicles was observed near the nucleus. Pinocytotic vesicles were seen arising from the membrane lining the flagellar pocket. They are associated with some microtubules which originate in the flagellar pocket region and extend toward the multivesicular structure. Morphometric analysis made on electron micrographs showed a mitochondrial relative volume of 0.11 and a peroxisome relative volume of 0.08. Determination of the number of sub-pellicular microtubules in different regions of the protozoon body show that the largest number is found in the region containing the Golgi complex. Carbohydrates were detected using the periodic-acid-thiosemicarbazide-silver proteinate technique. Reaction products were seen in the plasma membrane, in the membrane of the Golgi complex, and in the membranes which form the multivesicular structure. Two cytochemical methods were used to locate basic proteins. Using the ammoniacal silver method reaction products were seen only in the nucleus. With the ethanolic phosphotungstic acid method reaction products were seen in the microtubules which form the flagellum, in the peroxisome-like organelle, and at the region of adhesion of the flagellum to the cell body.     

Effect of Parmidine (pyridinolcarbamate) on platelet aggregation,blood coagulation,and fibrinolysis

The effect of Parmidine (pyridinolcarbamate) on platelet aggregation, blood coagulation, and fibrinolysis was studied in rabbits. After direct addition of Parmidine to platelet-enriched plasma aggregation of the platelets produced by serotonin (5-HT) and adrenalin was reduced. After oral administration of Parmidine to animals, aggregation induced by ADP, 5-HT, and adrenalin was inhibited, blood coagulation was reduced, and fibrinolysis was accelerated.Laboratory of Pharmacology, S. Ordzhonikidze All-Union Pharmaceutical Chemical Research Institute. Department of Pharmacology, Therapeutic Faculty, Moscow Medical Stomatological Institute. Translated from Byulleten' Éksperimental'noi Biologii i Meditsiny, Vol. 81, No. 3, pp. 322–324, March, 1976.     

Ultrastructure of the spermatozoon of Raillietina (Raillietina) baeri (Cyclophyllidea, Davaineidae) an intestinal parasite of the multimammate rat, Mastomys huberti (Rodentia, Muridae)

The mature Raillietina (Raillietina) baeri spermatozoon exhibits an apical cone of electron-dense material about 2.5 m long and 0.5 m wide and two helicoidal crest-like bodies roughly 100–125 nm thick. The latter are of different lengths, spiralized and stand in an angle of about 50° with the spermatozoon axis. The axoneme is of the 9 + 1 pattern and does not reach the posterior extremity of the gamete. The nucleus is an electron-dense cord coiled in a spiral around the axoneme. The cytoplasm exhibits a posterior densification and contains few small electron-dense granules in regions I, II and V of the spermatozoon. In regions III and IV, it is divided into irregular compartments by walls of electron-dense material. The cortical microtubules are spiralized and make an angle of 40–50° to the spermatozoon axis. In this work, we describe, for the first time, a spermatozoon of a davaineidaean cestode parasitic of mammals. This has enabled us to show a wide apical cone, which has never been described before in a cyclophyllidean species the spermatozoon of which has two crest-like bodies.