A new species of Eutyphoeus earthworm (Clitellata: Acanthodrilidae) from Mizoram, North-East India.

We describe here a new species of terrestrial earthworm Eutyphoeus rihnimensis sp. nov. (family Acanthodrilidae) from specimens collected at Khawrihnim, a village in Mizoram, Northeast India. The new species is distinguished from its congener members in E. gigas group in having an outgrowth that branches into four finger-like projections from the male porophores, five pairs of supra-intestinal glands in 8188 segments, typhlosole that begins from segment 25, ventral intestinal caeca 10 that covers segments 33-34, a bidiverticulate spermathecae with a short and stout duct, and very short lateral and median diverticula among other characters. Sequence analysis based on mitochondrial cytochrome c oxidase subunit 1 (CO1) gene further warrants its distinction from known species of Eutyphoeus.

Phytochemical analysis and in vitro anthelmintic activity of Imperata cylindrica underground parts.

BACKGROUND: Imperata cylindrica is considered as an agricultural weed, but it is a valuable plant in the traditional medicines of Southeast Asia. In the Mizo traditional medicine of India and Myanmar, the rhizomes and roots are used as a remedy for bacterial, fungal and intestinal helminth infections. METHODS: An extract of the whole underground parts was prepared in Soxhlet apparatus using chloroform as a solvent. After concentrating in a vacuum rotary evaporator, the extract was analysed using gas chromatography-mass spectrometry. Anthelmintic activity was tested in vitro against the tapeworm Raillietina tetragona and the roundworm Ascaridia galli. Scanning electron microscopy was used to examine the structural changes on the helminths after treatment with the plant extract. RESULTS: Twenty-two compounds were identified from the plant extract out of which fatty acids were the predominant compounds. Palmitic acid was the most abundant. Bioactive phytosterols such as campesterol and stigmasterol were also detected. The plant extract was significantly effective on both the helminths and showed dose-dependent anthelmintic activity as that of albendazole. The tapeworm treated with the plant extract showed deformities on the suckers, clumping of the spines, tegumental folds and erosion of microtriches. Extensive damage was also seen on the roundworm including cuticular shrinkage, collapse of the lips, and formation of warty surface throughout the body. CONCLUSION: I. cylindrica extract effectively killed and caused detrimental effects on parasitic tapeworm and roundworm. The study therefore validates the traditional usage among the Mizo people, and guarantees further investigation on the exact compound(s) and mechanism of action.

Anthelmintic activity of praziquantel and Spilanthes acmella extract on an intestinal cestode parasite.

Spilanthes acmella Murr., popularised as toothache plant, is a well-known culinary and medicinal plant for different purposes, but its use as an anthelmintic is apparently exclusive to the Mizo people of India and Myanmar. A chloroform extract of Spilanthes acmella Murr. was analysed in a single quadrupole GC-MS system, from which it was found that the major compound was an alkylamide, N-isobutyl-(2E,4Z,8Z,10E)-dodecatetraenamide. A comparative study was performed on the anthelmintic activity of the plant extract and praziquantel (PZQ) against an intestinal cestode, Raillietina echinobothrida. In terms of efficacy, PZQ was more potent, but the plant extract was also effective at all concentrations tested. PZQ caused severe shrinkage and folds of the tegument, constriction of the suckers, dislocation of spines and erosion of microtriches. The plant extract caused shrinkage and folds on the main body but not on the scolex. Damage on the suckers is more pronounced than in PZQ-treated cestodes. The spines were completely removed. The current findings indicate that S. acmella is a good source of compounds with anthelmintic activity.

Chemical composition and broad-spectrum anthelmintic activity of a cultivar of toothache plant, Acmella oleracea, from Mizoram, India.

Context: A variety of Acmella oleracea (L.) R.K. Jansen (Asteraceae) is used by the Mizo people of India and Myanmar for intestinal helminthiasis.Objective: To perform a chemical analysis of the plant extract using gas chromatography-mass spectrometry (GC-MS) and test the anthelmintic activity on intestinal parasites.Materials and methods: An extract of the aerial parts was prepared in hexane and analysed using GC-MS. Survival test was performed in vitro on the cestode, Taenia tetragona, and the nematode, Ascaridia perspicillum. Concentrations of 1.25, 2.5, 5, 10 and 20 mg/mL, prepared in phosphate-buffered saline (PBS) with 1% dimethylsulphoxide (DMSO), were tested. Negative control was maintained in PBS with DMSO, and albendazole was used as a reference drug. Each treatment consisted of six worms and was done until death was confirmed. Scanning electron microscopy was used to describe the structural changes.Results: Nineteen compounds were detected. The major compounds were fatty alcohols such as 3,7,11,15-tetramethylhexadec-2-en-1-ol and (9Z)-9-hexadecen-1-ol. Important bioactive compounds including an alkylamide, N-isobutyl-(2E,4Z,8Z,10E)-dodecatetraenamide, and a triterpenoid, lupeol, were also confirmed. The lethal concentration (LC50) of the plant extract was 5128.61 ppm on T. tetragona and 8921.50 ppm on A. perspicillum. Tegumental shrinkage, erosion of microtriches, and distortion of the suckers were observed on the cestode. The nematode showed collapse of the lips and shrunk cuticle.Conclusions: Acmella oleracea contains important bioactive compounds, which are responsible for the broad-spectrum anthelmintic activity. Further study on the pharmacology of the compounds is warranted.

Intestinal cestodes of chicken are effectively killed by quinoline-rich extract of Spilanthes acmella.

BACKGROUND AND AIM: Spilanthes acmella is used for the treatment of intestinal helminth infections in Mizo traditional medicine. In spite of a variety of drugs developed for helminthiases, an entirely safe and absolutely effective drug is still lacking, so much so that infections remain a major problem in human and animal welfare. In this study, we attempted to substantiate S. acmella as an anticestodal agent. MATERIALS AND METHODS: The aqueous extract of the aerial parts of S. acmella was prepared and from it a bioactive fraction was obtained using column chromatography. Chemical analyses were done using thin-layer chromatography (TLC) and gas chromatography-mass spectrometry (GC-MS). Helminth survival test was performed in vitro on an intestinal cestode, Raillietina tetragona. Structural effects on the cestode were examined under scanning electron microscopy. RESULTS: From the bioactive fraction of S. acmella extract, TLC indicated the presence of an aromatic quinone, which was identified using GC-MS as a quinoline derivative (2,2,4-trimethyl-1,2-dihydroquinoline having a retention time of 24.97 min and chemical formula of C12H15N). The quinoline-rich fraction showed concentration-dependent activity against R. tetragona as that of albendazole. Scanning electron microscopy of the treated cestode revealed classic anthelmintic effects such as tegumental shrinkage and damage of surface organs. The scolex was shrunk, suckers were degenerated with disintegrated spines, and rostellum was completely collapsed. There were severe damages on the tegument and formation of pit-like scars on the proglottids. CONCLUSION: The efficacy of S. acmella extract and structural damages it caused on the cestode indicates that it is a potential source of anthelmintic agent and that 2,2,4-trimethyl-1,2-dihydroquinoline contributes to its antiparasitic activity.