Anthelmintic efficacy of Holarrhena pubescens against Raillietina spp. of domestic fowl through ultrastructural, histochemical, biochemical and GLCM analysis

Holarrhena pubescens is a well-known medicinal plant used in ethno-traditional medicine and tribal folklore medicine in West Bengal, India. This article focuses on depicting the anthelmintic efficacy of ethanolic extract and Ethyl acetate fraction of the stem bark of Holarrhena pubescens against the cestode Raillietina spp. through efficacy testing, ultra-structural observations, histochemical and biochemical studies on some tegumental enzymes i.e., Acid Phosphatase (AcPase), Alkaline Phosphatase (AlkPase), Adenosine Triphosphatase (ATPase) and 5-Nucleotidase (5-Nu) along with Gray Level Co-occurrence Matrix (GLCM) analysis of histochemistry images. Praziquantel was used as the reference drug. Investigations revealed 10mg/ml of crude extract was determined as the most efficacious dose and the Ethyl acetate fraction showed the most anthelmintic property. Ultrastructural studies through Scanning Electron Microscope (SEM) and Transmission Electron Microscope (TEM) clearly showed the damage in head, sucker, proglottids, proximal and distal cytoplasm (DC), microtriches (MT), basal lamina (BL), nuclear membrane (NM), and, nucleolus (NL) in the treated worms. Histochemical studies revealed decrease in staining intensity for all the tegumental enzymes in the treated worms compared to control. The GLCM analysis strongly supported the result of histochemical studies. Biochemical studies revealed marked reduction in enzyme activity in the treated worms with maximum reduction in the activity of 5-Nu (77.8%) followed by ATPase (63.17%).