Sperm morphometry is the tool that confers objectivity to the morphological evaluation by accurately measuring the dimensions of the gamete and its structures. Thus, the aim of the study was to perform a morphometric characterization of the domestic cat sperm. Therefore, sperm samples were collected from twenty pairs of epididymis in a TRIS extender at 37ºC. An aliquot of the sample was used to make a smear with Rose Bengal solution, and afterwards, the morphology and morphometry were analysed. In the morphology, were quantified the percentage of normal sperm cells, morphological changes of head, midpiece and tail. In morphometry, each normal sperm cell was measured for length, width, area and perimeter of head and midpiece, tail length and total length. The parameters ellipticity, elongation, regularity and rugosity were also determined. The percentage of normal sperm was 67.21%. Of the abnormalities, the curled/folded tail, followed by the curved midpiece, abnormal shaped head and detached head were the most quantified. The sperm head presented 5.56 ± 0.01 μm and 3.10 ± 0.01 μm of length and width, respectively. The head area was 16.94 ± 0.05 μm2, while the perimeter was 16.16 ± 0.03 μm. In the derived parameters, the values were as follows: ellipticity of 1.81 ± 0.00; elongation of 21.39 ± 0.12; regularity of 0.81 ± 0.00; and rugosity of 0.14 ± 0.00. The midpiece presented length and width of 7.96 ± 0.01 μm and 0.76 ± 0.01 μm, respectively. The mean length of the sperm tail was 45.12 ± 0.06 μm, and the total cell size was 58.67 ± 0.06 μm. Thus, it was concluded that the cat sperm is an elongated cell, with high rugosity and regularity. The spermatic tail represents more than ¾ of the total length of the cell and the midpiece exceeds the length of the head.

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Background: Animal models are widely used in scientific research because of the ability to generate information from an organism like everything under a given experimental condition. Hematological and biochemical tests in laboratory animals are essential for the validation of several scientific studies. In addition, it standardizes physiological values   for these animals according to their sex, age, lineage, environment, and nutritional status. The present work aims to establish reference values   for biochemical and hematological standards in Balb/c mice, for males and females.Materials, Methods & Results: A total of 50 male and female mice were used at reproductive age. The procedures for collecting, processing, and analyzing the samples were standardized.  The collected blood samples were immediately transferred to eppendorf tubes containing heparin, and intended for hematological and biochemical evaluation. The hematological evaluation consisted of Red blood cell count (RBC), Leukocyte counts (WBC), Platelet counts (PLT), Hematocrit (HCT), Hemoglobin concentration (HGB), Mean corpuscular volume (MCV), and Mean corpuscular hemoglobin concentration (MCHC). Already the quantified biochemical parameters were: urea, creatinine, alanina aminotransaminase (ALT), aspartato aminotransaminase (AST) and fosfatase alcalina (FAL). The differential leukocyte count was also performed. Hematological results obtained for males and females were: 9.19 ± 3.35 (106/mm³) and 7.3 ± 2.01(106/mm³) of RBC; 35.8 ± 6.7% and 38.44 ± 3.93% of HCT; 11.51 ± 2.17 g/dL and 11.85 ± 1.56 g/dL of HGB; 45.83 ± 15.03 fL and 60.26 ± 18.25 fL of VCM; 31.80 ± 1.15% and 31.88 ± 0.99% of MCHC; and, 5380 ± 1994.21(10³/mm³) and 3564 ± 1071(10³/mm³) of WBC.  The platelet counts were 878.92 ± 84.19 and 678.28 ± 227.21, for males and females respectively. And for differential leukocyte counts, for males and females: eosinophils 2.12 ± 1.09% and 2.16 ± 1.71%; monocytes 2.84 ± 1.03% and 2.68 ± 1%; lymphocytes 68 ± 8.36% and 71.76 ± 5.9%; neutrophils 27.04 ± 8.55% and 22.96 ± 5.54%. Basophils were not quantified in the samples. As for the biochemical parameters, values of 54.16 ± 27.8 UI/L and 29.72 ± 4.4 UI/L of  ALT; 89.56 ± 47.73 UI/L and 71.32 ± 8.12 UI/L of AST; 3.76 ± 2.08 UI/L and 2.32 ± 0.85 UI/L of  FAL; 31.76 ± 21.08 mg/dL and 41.48 ± 13.61 mg/dL of urea; and 0.76 ± 0.18 mg/dL and 0.44 ± 0.11 mg/dL of creatinine.Discussion: The mean corpuscular volume, mean corpuscular hemoglobin concentration, leukocyte, and platelet counts diverged from those   found in literature. For the biochemical values, it was observed that creatinine values were different from those exhibited by other authors. Such divergences might be explained by the activity of endocrine organs, such as the production and/or release of activation/differentiation factors, and stress, applied methodology, lineage, or individual variability.  In addition, differences in the methodologies applied may be responsible for variations in hematological and biochemical values, requiring the standardization of the equipment and reagents used, as well as the adoption of a range that represents the minimum and maximum values within the normal physiological standard for given mouse lineage. In conclusion, the values presented in the present work are within the variation curve for rodents, and can be used as reference for other studies that use these animals.

Morphological sperm evaluation supported by the morphometry can be used in the determination of the seminal quality and in the investigation of potential extenders. Although there are studies comparing TRIS and ACP extenders, there are no comparative studies between them for the computerized assisted semen analysis (CASA), sperm viability, membrane functionality and sperm morphometry parameters of cryopreserved canine semen. Hence, we aimed to evaluate the effects of ACP-106c and TRIS on post-freezing canine sperm quality. Five dogs were submitted to semen collection twice with one-week interval. The semen was evaluated within the parameters: total motility, vigor, concentration, viability, plasma membrane functionality, morphology and morphometry. In the morphometric evaluation, the morphologically normal sperm was measured as: length, width, area and perimeter of the head and the midpiece, tail length and total length. The parameters of ellipticity, elongation, regularity and roughness were determined. Then, the semen was divided into two aliquots that were diluted in TRIS or ACP-106c, with the addition of egg yolk and glycerol. The diluted semen was refrigerated and frozen. The thawed samples were evaluated. Total motility, viability, sperm membrane functionality and normal morphology reduced after thawing in both extenders (morphology reduced from 89.60 ± 1.3% to 84.40 ± 1.8 and 84.60 ± 1.1% in TRIS and ACP-106c, respectively). However, it did not differ between TRIS and ACP-106c. In the ACP-106c the sperm head defects in cryopreserved semen were higher compared to fresh semen (P < 0.05). For all the morphometric parameters evaluated, there were no differences between fresh and cryopreserved samples (3.70 ± 0.4% vs. 2.30 ± 0.5%). In kinetics, with an interval of one week statistical differences between the extenders were found only in the parameters ALH and LIN (P < 0.05). Regardless of the extender, there were no changes in the morphometric parameters of sperm after thawing.

RESUMO:O cultivo de células animais existe há cerca de 100 anos. Os estudos pioneiros de pesquisadores como Harrison, Carel e Roux, contribuíram para o desenvolvimento da técnica e para encorajamento de outros trabalhos relacionados ao cultivo de células. Cultivo pode ser definido como um conjunto de práticas que permitem a manutenção de células em um sistema in vitro independente do tecido de origem sob condições controladas. Os primeiros cultivos resumiam-se a tecidos fragmentados submersos em plasma sanguíneo (ou soro do animal de origem) contidos em frascos. Atualmente, o cultivo de células apresenta inúmeras aplicações, desde a produção biotecnológica de moléculas recombinantes, anticorpos monoclonais, vacinas veterinárias e humanas, à produção de enxertos para transplantes. Muitos desses avanços só foram possíveis graças ao estabelecimento de linhagens celulares estáveis e a descoberta de sistemas de cultivo específicos para cada tipo celular. Portanto, o presente artigo tem como finalidade apresentar um apanhado histórico do cultivo de células animais, além de expor as linhagens celulares de referência mundial.Palavras-chave: cultivo, linhagem, bioengenharia, biotecnologia. History of development of the cultivation animal cells.ABSTRACT: The cultivation of animal cells has been around for 100 years. The pioneering studies by researchers such as Harrison, Carel and Roux, contributed to the development of the technique and encouragement to other works related to cell culture. Cultivation can be defined as a set of practices that allow the maintenance of cells in an in vitro system independent of the original tissue under controlled conditions. The first cultures are summarized to fragmented tissue submerged in blood plasma (or serum of animal origin) contained in flasks. Currently, the cell culture has numerous applications from biotechnological production of recombinant molecules, monoclonal antibodies, human and veterinary vaccines, to the production of grafts for transplantation. Many of these advances have been possible only through establishing stable cell lines and the discovery of specific culture systems for each cell type. Therefore, this article aims to present a historical overview of animal cell culture, showing the future expectations of this technology, and expose worldwide reference cell lines.

This study evaluated the effect of the extract of Aloe vera at concentrations of 10% and 20% on the cryopreservation of sperm from the epididymis of domestic cats. Epididymal spermatozoa were recovered using the flotation technique and used in the treatments: control (TRIS-egg yolk at 20%), T10% (TRIS plus 10% of A. vera extract), and T20% (TRIS plus 20% of A. vera extract). The spermatozoa were subjected to 4ºC for 60 minutes, followed by 20 minutes in nitrogen vapors, and stored in a cryogenic cylinder. The samples were thawed at 37°C for 30 seconds. The sperm motility decreased (P<0.05) after thawing in the three treatments. Only the spermatozoa in the control treatment maintained postthawing vigor. The viability of spermatozoa decreased in the treatments with A. vera (P<0.05). According to the hypoosmotic test, all treatments maintained the sperm membrane functionality (P>0.05) during freezing; however, after thawing, it decreased (P<0.05) in the T10% and T20% treatments. The morphology and chromatin condensation of spermatozoa did not differ, regardless of the treatments and time of evaluation (P>0.05). The effect of the crude A. vera extract was not satisfactory on the cryopreservation of epididymal spermatozoa of domestic cats after thawing; although the motility of spermatozoa was similar to that found with the use of egg yolk, and it presented maintenance of the chromatin integrity. However, it is necessary to understand the action of the substances present in A. vera with the feline spermatozoa, well as the standardization and adjustment of physicochemical characteristics aiming at the future application of the vegetal extract.