Using specific skeletons staining with ALIZARINe red S (AR) and alcian BLUE (AB) which is a significant method for of ossification centers during embryogenesis. One major problem in this method was been percipitation of chemical stain which has been resulted in weak to no reaction in different stages of bone and cartilage development. The purpose of this study was to develop new way to over come this technical problem. By using high concentration of AR (2.5 volume) combined with less concentration AB (0.5 volume) as well as decreasing processor time from 3 days to 1.5 and also we decreased the acidity of staining solution up to pH 5, we were able to show that after clearing of embryos, the soft tissues are absolutely clear and cartilages and bones were stained perfectly. It seems that with modify processors, dye can penetrate the embryonic tissues and remain in place after penetration. As high acidity may decreased the amount of bone, it is recommended to reduced the rate of acidity.

Normal and abnormal developmental studies on chicken embryo system are necessart, as the widespread use of chicken embryo model in embryological experiments, from organ to cell and molecule levels. Knowledge of normal embryonic development is important to understanding embryonic events and efficient performing of experimental and teratological tests. The present study was undertaken by double staining on chicken embryo from 1st to 21st day of incubation period by ALIZARIN red – alcian BLUE double staining method, to investigation of wing and leg skeleton development. In this method, cartilage and bone matrix stain BLUE and red, respectively. Totally 13 pieces bone in wing and 22 pieces in leg skeleton were determined. Then, the time of cartilage formation and their ossification, were detected all of those become ossified through bones, become ossified through enchondral ossification pathway, but some part of each bone remains cartilaginous. Results show that leg skeleton development is earlier than wing skeleton development. 6th day and 11th day of incubation period are first appearance cartilage and bone matrix, respectively. These skeletogenous stages of the chicken embryo wing and leg will be useful as a normal control not only in studies of experimental embryology and teratology, but also in the analysis of mutant embryos with skeletal abnormalities.

Introduction: Skeletal staining is an important part of anatomical studies which can evaluate macroscopic disorders in bones and cartilages development. The aim of the present article is to illustrate two distinct protocols by all details in order to stain skeletal system of the lab animals in all ages of their lives so it becomes easy for future studies in this field.Methods: In this paper, staining method was based on sample’s ages. So, all the steps of double skeletal staining described in 2 protocols, one for embryos and newborns and another for adults. Fixing by ethanol, alcian BLUE staining, dehydration by ethanol, ALIZARIN red staining, washing by potassium hydroxide and glycerol, storage in pure glycerol were the stages of embryos and newborns skeletal staining. Fixing by neutral formalin buffer, placing the samples in ddH2O, fixing again by ethanol, skinning and eviscerating, alcian BLUE staining, trypsin digestion, ALIZARIN red staining, potassium hydroxide clearing, KOH and glycerol clearing and finally storing the samples in pure glycerol were the steps of adults staining.Results: Outcomes of a good and reliable procedure for skeletal staining with ALIZARIN red and alcian BLUE are the samples whose cartilages and bones are stained BLUE and red respectively. They are completely transparent so that the skeletal parts can be seen through them.Conclusion: It is shown that skeletal staining procedures differ in embryos and adults although in the most surveys were tried to use a same protocol in diferent ages of laboratory animals.

Studying skeletal system in embryonic and after birth periods has been focused by researchers from different fields, biologists, embryologists and archaeologists, for a long time. This study can be useful in normal embryology and fatal disorders researches, as well as skeleton development system. This research may lead to experimental teratological studies. This paper studies enchondral ossification. in chicken embryo's long bone as a laboratory model by microscopic and macroscopic methods, which later analyzed by ALIZARIN red Alican BLUE double staining method. Results showed that, unlike mammals, birds' cartilage change to bone in four continuous stages including: resting phase, proliferative, hypertrophy and ossification phase. The difference is that there is no calcification phase in birds. Studying the samples prepared by ALIZARIN – Alican method shows that cartilage bone matrix become BLUE and red .respectively. We demonstrated that there is a gap between cartilage and bone which remains without color. This gap is hypertrophic phase that was confirmed by microscopic method.

This study investigated the adsorption of ALIZARIN red (AR) from aqueous solutions using Epichlorohydrin-Modified Walnut Shells (EMWS). Through static adsorption experiments, the impact of contact time (tc), initial pH of the solution, solution concentration (c0), and temperature (T) on the adsorption of AR onto the EMWS was examined. The optimum values of tc and pH were determined to be 360 min and 3.51, respectively. The adsorption capacity of the EMWS for AR increased when c0 and T were increased. Non-linear fitting of the equilibrium data was performed using the Langmuir, Freundlich, and Temkin isotherms. AR adsorption was well explained by the Langmuir isotherm with the qm of 81.44 mg/g at 323 K. To simulate the adsorption kinetics, various models including pseudo-first-order, pseudo-second-order, intra-particle diffusion, and Elovich models were employed. The results indicated that it followed the PSO kinetic model (k = 4.56 × 10 -4 g / (mg·min), qe = 18.84 mg/g and R2 = 0.9808). Moreover, the Elovich model confirmed the occurrence of chemisorption. According to thermodynamic investigations, the adsorption of AR is found to be spontaneous (ΔG﹤0) and endothermic (ΔH = 12.24 kJ/mol) while exhibiting an increase in entropy (ΔS = 58.5 J / (mol.K)). The results indicated that EMWS showed great potential as an affordable and efficient adsorbent for treating AR wastewater.

Introduction & Objective: The traditional method for staining cartilage and bone in vertebrates is to use Alcian BLUE and ALIZARIN red. Acid is used in the base of the ingredients of Alcian BLUE solution, which led to clear distinguishing of the stained skeletal structures as much as possible. But the problem is that if acid is used, some of the bone ions are removed leading to errors in the results. Therefore, the present study aimed to compare two using acid and acid-free protocols for staining cartilage and bone during the early development of Common bream (Abramis brama) to provide a solution in future studies. Material and Methods: For staining, we applied acid-based (acetic acid) and acid-free protocols using Alican BLUE and ALIZARIN red during the processing stages of watering, staining of cartilage and bone, neutralization, watering, protein digestion, color removal, and washing and clearing. Results and Discussion: Based on the results, it can be stated that in studies of the early development of bony fishes, the use of cartilage and bone staining method based on the acid-free protocol is more suitable than using acid because this method helps to better understand the sequence and pattern of skeletal ontogeny more accurately during the early development period.

Coating metal with a polymeric film is the most widely employed protection procedure against the corrosion of metallic substances. However, this coating can be damaged by microbiological species. The existence of porous layer makes the oxide highly absorbent and hence suitable to be painted. It is well known that biofilms formation and bacteria activity can be influenced by the interaction between metal surfaces and the existing dye. Conventional electrochemical techniques provide valuable information about these processes which are electrochemical in nature. In this paper, the resistance effect of an organic dye (ALIZARIN, 1, 2-dihydroxyanthraquinone) on anodic oxide film of anodized aluminum plates against MIC was studied using various electrochemically methods including: Cyclic and ac voltammetry and scanning electron microscopy (SEM). The results showed, that this dye decreased the growth of bacteria, on dye coated, protective layer on the surface of porous oxide film of anodized aluminum.

A total of 610 strains from 16 species of micromycetes with potential ligninolytic capabilities were isolated from soil, compost, and rotten wood, including 400 strains of soil origin. In the test of decolorization of agarized 0.06% ALIZARIN BLUE, 52 strains with high decolorization activity were selected. In liquid cultures with 0.2% post-industrial lignin and 0.03% ALIZARIN BLUE, all of the selected strains showed three different extracellular peroxidase activities such as HRP-like, LiP, and MnP. Laccase was detected only in the cultures with ALIZARIN BLUE, but those were trace amounts. In the decolorization of ALIZARIN BLUE, the highest levels of activity were attained by HRP-like peroxidase. In the transformations of post-industrial lignin that included decolorization, colorization (darkening of substrate), and re-colorization, the highest levels of activity were attained by lignin peroxidase, especially at the start of the culture. The transformations of both dye substrates in the cultures of all 52 strains were accompanied by the release of glucose oxidase, but high activities of that enzyme were observed only in the presence of ALIZARIN BLUE. Whereas in the presence of lignin the fungi under study synthesized catalase, the activity of which attained a maximum toward the end of the culture was negatively correlated with the activity of the remaining oxidoreductases under study. Among the 52 selected strains, three strains such as Haematonectria haematacocca BwIII43, K37, and Trichoderma harzianum BsIII33 were characterized by a high activity of decolorization.