RESUMO
Background: Varanus salvator is one of the reptiles being hunted by human beings for several purposes, including traditional medicine. The studies about reproductive biology aspects were limited. Aim: This study aimed to determine the morphology, histology, and histometry of V. salvator paryphasmata and hemibaculum based on Snout-Vent Length (SVL) as an indicator of sexual maturity. Methods: This study examined 18 pairs of hemipenis of V. salvator with SVL more and less than 40 cm in equal number. Paryphasmata and hemibaculum parts were observed visually and micro-sliced, then stained with Hematoxylin-Eosin (HE). The histological observation was conducted under a 40×, 100×, and 400× magnification of a light microscope. The histometry of the paryphasmata was examined using 13 Megapixels Coolpad and OptiLab Plus for microscopic pictures. The chondrocyte cell area was measured using the Optilab Plus and Image Raster three applications. Results: The sizes of glans of hemipenis, paryphasmata, and hemibaculum increased according to the increasing of SVL. The average paryphasmata row number, epidermis, and loose connective tissue thickness were not significantly different (p > 0.05). However, dense connective tissue was thicker (p < 0.05), which corresponds to SVL. Hemibaculum was composed of fibrous and hyaline cartilage characterized by chondrocyte cells. The SVL also affects (p < 0.05) the ossification of hyaline in hemipenis, while the chondrocyte cell area followed the equation -1.87E7 + 7.09E5* SVL. Conclusion: The SVL size of V. salvator affects the paryphasmata, hemibaculum, thickness of dense connective tissue of paryphasmata, and the area of chondrocyte cells.
Assuntos
Lagartos , Animais , Condrócitos , HumanosRESUMO
Fixation is one of the processes in preparing histology and pathology. The common material for fixation is buffered formalin including paraformaldehyde. However, the effect of the damaged cells, which is fixed for a long time, causes the research for other fixation materials to become necessary. In addition, paraformaldehyde is also harmful to human body and natural environment. Ethanol is one of the alternative fixation materials, which has been used for two hundred years. It has been used for many purposes, both in routine staining and immunohistochemistry. Nonetheless, no research confirms its effect on the electron microscope. The authors studied the effect of 50 % of ethanol on the cell membrane, organelles, and nucleus of Purkinje cells (Neuron purkinjense) observed on a light microscope and Transmitted Electron Microscope (TEM). Then it was compared to buffered formalin. In the light microscope, it shows that both of fixations have no different effects of the morphology of the cell membrane, cytoplasm, the nucleus of Purkinje cells and the neutrophils. We assume that our 50 % of ethanol concentration is almost the same as BF 10 % in the ability of hardening tissue and color absorption based on the previous study. In TEM, the structure of the cell membrane, organelles, and cytoplasm of Purkinje cell look broken in the cerebellum of 50 % of ethanol except for the nucleus. There was no significant difference diameter of the nucleus. It happened in general because of the shrinkage effect of ethanol. However, the authors recommend using 50 % of ethanol for routine staining.
La fijación es uno de los procesos en la preparación de muestras para histología y patología. El material más común para la fijación es la formalina tamponada. Sin embargo, el daño a las células que se mantienen en formalina durante mucho tiempo, hace necesario buscar otros materiales de fijación. Además, el paraformaldehido también es perjudicial para el cuerpo humano y el medio ambiente natural. El etanol es uno de los materiales de fijación alternativos que se ha utilizado durante muchos años, con diversos objetivos, tanto en la tinción de rutina como en la inmunohistoquímica. Sin embargo no se ha confirmdo su efecto con microscopio electrónico. Los autores estudiaron el efecto del 50 % de etanol sobre la membrana celular, los orgánulos y el núcleo de las células de Purkinje observados en un microscopio óptico y un microscopio de transmisión electrónico (TEM). Luego se comparó con la formalina tamponada. En el microscopio óptico se observó que ambas fijaciones no tienen efectos diferentes a la morfología de la membrana celular, el citoplasma, el núcleo de las células de Purkinje y los neutrófilos. Suponemos que nuestra concentración de 50 % de etanol es casi la misma que BF 10 % en la capacidad de endurecer el tejido y la absorción de color según el estudio anterior. En TEM, la estructura de la membrana celular, los orgánulos y el citoplasma de la célula de Purkinje presentaban daño en el cerebelo con un 50 % de etanol, a excepción del núcleo. No hubo diferencia significativa en el diámetro del núcleo. En general lo anterior se debió al efecto de contracción del etanol. En conclusión los autores recomiendan usar 50% de etanol para la tinción de rutina.