Evaluation of in vitro anthelminthic effect of crude leaves extract of piper betle L. (sireh) on commonest gastrointestinal helminths obtained from goats

Abstract

Livestock industry such as goat farming has been providing the farmers in developing countries with steady income. However, farm mismanagement, poor welfare and climate factor has contributed to the increase infections of helminths in goats which challenged the demands-supply chain. The condition worsened when they showed signs of resistance towards majority of commonly available anthelmintic drug. Improving animal husbandry and plant-based treatment could become an alternative in treating helminths infections in goats. Thus, the study is aimed to identify the occurrences of gastrointestinal helminths in goats’ faecal samples and determines the anthelmintic potential of Piper betle L. Following the physical screening, 120 goats’ faecal sample from three different farms in Kuantan, Pahang were observed for helminths under microscope. Subsequently, the species of helminths were confirmed using real-time PCR and high-resolution melt assay (qPCR-HRM) and the predominant species was determined. Then, aqueous and methanol extracts from the Piper betle L. leaves that were prepared for the anthelmintic assay were screened for their phytochemical compounds using liquid and gas chromatography-mass spectrometry (LCMS) and (GCMS) respectively. In addition, their feed nutritional assessments were identified. Finally, the anthelminthic effect of Piper betle L. leaves extracts were examined on the predominant species using egg hatch assay (EHA) and larval paralysis assay (LPA). This study revealed 74.17% (89/120) samples of positive gastrointestinal helminths species. Haemonchus contortus was predominant among the six different species discovered. Besides, the study also revealed multiple infections of internal helminths in a sample. The diagnosis of Helminthiasis is often unrecognized in animal. It is usually suspected on the basis of physical inspection. However, this study has found no direct relationship between physical screening and microscopic observation. Goats with perfect body score were actually severely infected when their faecal samples were studied. Thus, the appearance of helminth under microscopic and further confirmation with molecular assay not only confirms the occurrence but also the species. Both GCMS and LCMS revealed ‘Phenols’ as majority bioactive compound that is responsible for the anthelmintic property. Nevertheless, LCMS has found many more compounds with different possible uses that could be utilised further. All treatment concentration of Piper betle L. has effectively induced anthelmintic activity. However, 100% mortality on the death of Haemonchus contortus eggs and larvae was successfully achieved at the concentration of 5.0 mg/ml within 24 hours for EHA and concentration of 7.5 mg/ml within 12 hours for LPA. In conclusions, Piper betle L. leaves indeed possess the anthelmintic properties where it successfully eliminated the predominant species of gastrointestinal helminths from goats. Nevertheless, these strategies represent possible therapeutic treatment in the future.