Comparison of cartilage and bone staining methods based on using acid and acid-free protocols for study of ontogeny in common Bream (Abramis brama)

Subject Areas :

Mohammad Reza Sahraein ¹ , ایگدری Eagderi ² , Gholamreza Rafiee ³ , Arash Zibaee ⁴ , Mohamad Moradi Ghrkhlo ⁵

1 - PhD graduate, Department of Fisheries, Faculty of Natural Resources, University of Tehran, Karaj, Iran.
2 - Department of Fisheries, Faculty of Natural Resources, University of Tehran, Karaj, Iran.
3 - Professor, Department of Fisheries, Faculty of Natural Resources, University of Tehran, Karaj, Iran.
4 - Associate professor, Department of Plant Protection, Faculty of Agricultural Sciences, University of Guilan, Rasht, Iran.
5 - Department of Biology, Faculty of Science, University of Zanjan, Zanjan, Iran.

Received: 2022-09-14 Accepted : 2022-11-02 Published : 2022-11-22

Keywords: larval development, Common bream, Alcian blue, Alizarin red, Osteology,

Abstract :

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.