Morphological, cytochemical and ultrastructural aspects of blood cells in freshwater stingray species in the middle Rio Negro basin of Amazonian Brazil

In the present work, we examined the morphology, dimensions, cytochemical staining reactions and ultrastructure of blood cells from three freshwater stingray species, Potamotrygon wallacei, Potamotrygon motoro and Paratrygon aiereba, living in the waters of the middle Rio Negro basin (Barcelos, Amazonas, Brazil). We identified erythrocytes, erythroblasts, thrombocytes and four types of leukocytes (basophils, heterophils, lymphocytes and monocytes) in the blood of these stingray species. In all the freshwater stingray species studied, the shapes and dimensions of these cells were similar to those of marine elasmobranchs. Positive PAS staining occurred in heterophils and thrombocytes, and weak staining occurred in lymphocytes and monocytes, while metachromasia only occurred in basophils. Positive Sudan Black B staining was observed in thrombocytes and lymphocytes, and weak staining occurred in heterophils. Basophils and heterophils were the only cells with positive bromophenol blue staining, while no peroxidase staining was observed in any of the four leukocyte types. This is the first study to establish the dimensions and cytochemical staining profiles of blood cells in Amazonian stingray species. Because these elasmobranch species are exported as ornamental fish to countries worldwide, this study can contribute to establishing standards for blood constituents that may be helpful in assessing the health and welfare of these fish in artificial systems.

Morphological, Cytochemical and Ultrastructural Aspects of Blood Cells in Freshwater Stingray Species in the Middle Rio Negro Basin, Amazonian, Brazil

Examined the morphology, dimensions, cytochemical staining reactions and ultrastructure of blood cells from three freshwater stingray species, Potamotrygon wallacei, Potamotrygon motoro and Paratrygon aiereba, living in the waters of the middle Rio Negro basin (Barcelos, Amazonas, Brazil). We identified erythrocytes, erythroblasts, thrombocytes and four types of leukocyte (basophils, heterophils, lymphocytes and monocytes) in the blood of these stingray species. In all the freshwater stingrays studied, the shape and dimensions of these cells were similar to those of marine elasmobranchs. A positive PAS reaction occurred in heterophils and thrombocytes, and a weak reaction in lymphocytes and monocytes, while a metachromasia reaction only occurred in basophils. Sudan black staining was positive for thrombocytes and lymphocytes, and only a weak reaction occurred in heterophils. Basophils and heterophils were the only cells stained with bromophenol blue, while no peroxidase reaction was observed in any leukocyte type. This is the first study to establish the dimensions and cytochemical staining reactions of blood cells in Amazonian stingray species. Since these elasmobranch species are exported as ornamental fish to countries worldwide, this study can contribute towards establishing standards for blood constituents that may be helpful in assessing the health and welfare of these fish in artificial systems.

Hematological and cytochemical characteristics of peripheral blood cells in the argus snakehead (Ophiocephalus argus Cantor)

Background The argus snakehead (Ophiocephalus argus Cantor) is a highly nutritious, freshwater, cultured bony fish with a high economic value. The health of the fish is closely related to its blood cells, which are critical for oxygen transport, natural defense, and immunity. We investigated the morphometry, microstructure, and cytochemical characteristics of the peripheral blood cells of O. argus. Our results may provide the basic reference values needed to monitor the health of this fish for large-scale cultivation. Methods The number of blood cells in O. argus were counted on a hemocytometer and their size was measured using a micrometer under light microscope. The morphology and classification of the blood cells were studied using Wright’s staining and the cytochemical characteristics were studied using seven chemical stains including peroxidase (POX), Sudan black B (SBB), periodic acid-Schiff (PAS), acid phosphatase (ACP), alkaline phosphatase (ALP), chloroacetic acid AS-D naphthol esterase (AS-D), and α-naphthol acetate esterase (α-NAE). Results The peripheral blood cells in O. argus can be classified as erythrocytes, leukocytes, and thrombocytes; of which, females had 2.9597 million/mm3, 88,400/mm3, and 43,600/mm3, respectively, and males had 3.0105 million/mm3, 105,500/mm3, and 34,000/mm3, respectively. Leukocytes consisted of neutrophils, monocytes, large lymphocytes, and small lymphocytes. Eosinophils and basophils were not found. Monocytes were the most numerous leukocytes identified, followed by neutrophils and small lymphocytes, while large lymphocytes were the least frequently identified. Cytochemical staining showed that erythrocytes were only positive for PAS staining. Neutrophils were strongly positive for POX, SBB, and ACP, and positive for all the other cytochemical stains. Monocytes were positive for PAS and α-NAE and were weakly positive for ACP and AS-D staining. Large lymphocytes were positive for PAS and were weakly positive for ALP, AS-D, and α-NAE staining. Small lymphocytes were positive for PAS and weakly positive for AS-D and α-NAE staining. Thrombocytes were positive for PAS and were weakly positive for ACP and AS-D, but negative for the remaining cytochemical stains. The morphology of peripheral blood cells in O. argus was generally similar to that of other fish species, while the cytochemical staining patterns showed clear species specificity.

Hematologic and Biochemical Reference Intervals and Urinary Test Results for Wild-caught Adult Southern Giant Pouched Rats (Cricetomys ansorgei)

Southern giant pouched rats (Cricetomys ansorgei) are muroid rodents native to subSaharan Africa. They are increasinglyused as service animals because of their keen sense of smell and are primarily known for clearing minefields in Africa. Theobjectives of this study were to determine hematologic and biochemical reference intervals from clinically healthy wild-caught captive adult rats, to describe the cytochemical staining reactions of peripheral blood leukocytes, and to document urinalysis findings. Blood samples were collected from the coccygeal artery of 60 isoflurane-anesthetized rats (36 males and 24 females) and analyzed with automated hematologic and biochemical analyzers; manual differential cell counts were performed on modified Wright–stained blood smears. Urine was collected by cystocentesis, and dipsticks were analyzed on a urine analyzer, with visual examination of unstained sediments. Samples from a male rat with chronic renal disease were excluded from analysis. Reference intervals were determined according to guidelines established by the American Society of Veterinary Clinical Pathology. Lymphocytes were the dominant leukocyte in peripheral blood and granular lymphocytes were identified in most animals. Male rats had significantly higher RBC, absolute reticulocyte counts, and MCV than did female rats. Minor sex-associated differences in urea nitrogen concentration and GGT activity were noted. Leukocytes showed unique cytochemical staining characteristics. Small amounts of protein and bilirubin were found in the urine of rats of both sexes and of female rats, respectively, particularly in concentrated urine. These results will provide benchmarks for determining health status and identifying disease in this species of rat.

Chondrogenic Progenitor Cells Exhibit Superiority Over Mesenchymal Stem Cells and Chondrocytes in Platelet-Rich Plasma Scaffold-Based Cartilage Regeneration

Background: Platelet-rich plasma (PRP) has been considered a promising tool for cartilage regeneration. However, increasing evidence has demonstrated the controversial effects of PRP on tissue regeneration, partially due to the unsatisfactory cell source. Chondrogenic progenitor cells (CPCs) have gained increasing attention as a potential cell source due to their self-renewal and multipotency, especially toward the chondrogenic lineage, and, thus, may be an appropriate alternative for cartilage engineering. Purpose: To compare the effects of PRP on CPC, mesenchymal stem cell (MSC), and chondrocyte proliferation, chondrogenesis, and cartilage regeneration. Study Design: Controlled laboratory study. Methods: Whole blood samples were obtained from 5 human donors to create PRPs (0, 1000 × 109, and 2000 × 109 platelets per liter). The proliferation and chondrogenesis of CPCs, bone marrow–derived MSCs (BMSCs), and chondrocytes were evaluated via growth kinetic and CCK-8 assays. Immunofluorescence, cytochemical staining, and gene expression analyses were performed to assess chondrogenic differentiation and cartilaginous matrix formation. The in vivo effects of CPCs, BMSCs, and chondrocytes on cartilage regeneration after PRP treatment were measured by use of histopathological, biochemical, and biomechanical techniques in a cartilage defect model involving mature male New Zealand White rabbits (critical size, 5 mm). Results: The CPCs possessed migration abilities and proliferative capacities superior to those of the chondrocytes, while exhibiting a chondrogenic predisposition stronger than that of the BMSCs. The growth kinetic, CCK-8, cytochemical staining, and biochemical analyses revealed that the CPCs simultaneously displayed a higher cell density than the chondrocytes and stronger chondrogenesis than the BMSCs after PRP stimulation. In addition, the in vivo study demonstrated that the PRP+CPC construct yielded better histological (International Cartilage Repair Society [ICRS] score, mean ± SEM, 1197.2 ± 163.2) and biomechanical (tensile modulus, 1.523 ± 0.194) results than the PRP+BMSC (701.1 ± 104.9, P < .05; 0.791 ± 0.151, P < .05) and PRP+chondrocyte (541.6 ± 98.3, P < .01; 0.587 ± 0.142, P < .01) constructs at 12 weeks after implantation. Conclusion: CPCs exhibit superiority over MSCs and chondrocytes in PRP scaffold-based cartilage regeneration, and PRP+CPC treatment may be a favorable strategy for cartilage repair. Clinical Relevance: These findings provide evidence highlighting the preferable role of CPCs as a cell source in PRP-mediated cartilage regeneration and may help researchers address the problem of unsatisfactory cell sources in cartilage engineering.

Peculiar traits of wood in a leaning stem of Scots pine (Pinus sylvestris L.)

The paper analyses biometrical and anatomical traits of wood in a leaning stem of pine trees. For study purpose, five pine trees (Pinus sylvestrisL.) with a visibly leaning stem were chosen. Wood samples were taken at three different stem heights, that is, below the stem curvature, at the curvature and above the curvature. Microscopic specimens were prepared and used for the following measurements: annual rings width, tracheids diameter and tracheids wall thickness. The measurements were performed for wood located on the lower side of the leaning stem and on the opposite side. Cytochemical staining was performed to identify the occurrence of laricinan. The results showed tracheids with a rounded shape and thick cell walls, helical cavities and intercellular spaces in wood located at curvature height on the lower side of the leaning stem. These traits indicate a severe compression wood that allowed pine trees to change their stem position in relation to the vector of gravity.