Abstract

Comparative Studies of Different Fixatives Used for Histopathology in Broiler Bird Tissues

Authors: Raheel Sharif, Muhammad Yasin Tipu, Ghulam Mustafa, Muhammad Rizwan, Kashod Aslam, Laraib Khan, Haziq Abdullah, Amina Yaseen and Asjad Umair Shah

DOI: http://dx.doi.org/10.71081/cvj/2024.029

Abstract
Formalin has been proven to be an effective fixative in ordinary tissue processing, but there are significant safety and health risks associated with its use. Formalin can be safely substituted with honey, jaggery, ethanol (70 %), and a mixture of ethanol, methanol and acetic acid (EMA=3:1:1). A total of 270 bits were collected from the liver, lungs, and intestines and fixed at different concentrations and durations. The aim of this study was to compare the tissue fixation properties of honey, jaggary, ethanol 70%, and EMA with those of formalin based on antibacterial properties, staining features of tissues, prolonged embedding of tissues in paraffin wax, and evaluation of nuclear staining, cytoplasm staining, cell morphology, clarity of staining, and uniformity of staining with H&E. This study found that honey, ethanol, EMA and formalin possess antibacterial properties and are effective fixatives for tissues. However, honey and jaggary showed poor fixation quality compared to EMA and formalin at a time duration of 24 h for liver and intestine tissues, while honey and jaggary gave comparable results to formalin, ethanol 70%, and a mixture of ethanol, methanol, and acetic acid (EMA) in the lungs. In addition, formalin and EMA caused tissue damage and shrinkage when embedding the tissue in paraffin wax for up to five hours, especially in the lungs. The study also found that the concentrations of honey and jaggary did not affect the staining quality of tissues, and a concentration of more than 50% could disturb the staining quality. These results suggest that natural fixatives such as honey and jaggary and chemical fixatives such as EMA (a mixture of ethanol, methanol, and acetic acid) could be considered as alternatives to formalin for tissue fixation in histopathological research.

Keywords: Histopathology, Broiler bird tissues.