scholarly journals In Vitro Anthelminthic Efficacy of Aqueous Pomegranate (Punica granatum L.) Extracts against Gastrointestinal Nematodes of Sheep

Pathogens ◽  
2020 ◽  
Vol 9 (12) ◽  
pp. 1063
Author(s):  
Fabio Castagna ◽  
Domenico Britti ◽  
Manuela Oliverio ◽  
Antonio Bosco ◽  
Sonia Bonacci ◽  
...  
Keyword(s):  
Gallic Acid ◽  
Anthelmintic Resistance ◽  
Punica Granatum ◽  
In Vivo Studies ◽  
Insoluble Residue ◽  
Methanol Fraction ◽  
Egg Hatch ◽  
Anthelmintic Efficacy ◽  
Egg Hatch Test

The worldwide increased difficulty to counteract gastrointestinal nematode (GIN) infection in sheep, due to progressing anthelmintic resistance, has led to the evaluation of other alternative helminth control options, mainly from plants. The anthelmintic efficacy of an aqueous Punica granatum macerate was evaluated in sheep naturally infected by GIN in southern Italy. The macerate was chemically characterized by chromatographic analysis coupled with high-resolution mass spectrometry (LC/HRMS) and an aliquot was concentrated to obtain a dry extract. A part was characterized, the remaining washed with methanol to obtain an insoluble residue and methanol phase. In the methanol fraction, the quantitatively predominant gallic acid was purified to obtain the pure molecule. The three fractions thus obtained were used for in vitro studies (i.e., egg hatch test) to verify anthelmintic efficacy. For this purpose, fecal samples were collected from sheep naturally infected by GINs. Fractions were diluted in H2O/DMSO 0.5% at 1.00, 0.5, 0.25, 0.125, 0.05, and 0.005 mg/mL concentrations. Thiabendazole (0.25 and 0.5 mg/mL) and deionized water were used as positive and negative controls, respectively. Egg hatch test results indicated that all fractions caused a significant (p < 0.05) egg hatch inhibition within 48 h of exposure highlighting a high (>82%) efficacy in vitro at all tested doses. Maximal egg hatching inhibition effect was exhibited by the methanol fraction (99.3% and 89.3% at 1 and 0.005 mg/mL concentrations), followed by the insoluble residue and gallic acid (94.7% and 85.3% and 94.0% and 82.7% at 1 and 0.005 mg/mL, respectively). The current study validated the anthelmintic potential of traditional P. granatum macerate against GIN infection in sheep, thus highlighting the role of gallic acid as principal component and justifying a need to undertake further in vivo studies on these ethno-veterinary remedies.

scholarly journals In Vivo and In Vitro Pharmacodynamics of Anthelmintic Medication Used in Sheep

2019 ◽  
Vol 76 (2) ◽  
pp. 175
Author(s):  
Mihai CERNEA ◽  
Roxana FILIP ◽  
Raul CĂTANĂ ◽  
Laura CĂTANĂ
Keyword(s):  
Anthelmintic Resistance ◽  
In Vivo Studies ◽  
In Vitro Tests ◽  
Minimal Risk ◽  
Egg Hatch ◽  
Superior Efficacy ◽  
Reference Concentration ◽  
Post Therapy

Our research aimed to evaluate the efficacy of anthelmintic treatments in sheep, as well as the resistance occurrence risk for the most commonly used substances. Anthelmintic medication efficacy was evaluated on 30 animals from a private farm, located in Sânmihaiu Almaşului, Sălaj County. In vivo studies were performed by using Fecal Egg Count Reduction Test (FECRT) and testing an albendazole-based (ABZ) product. In vitro, we used Egg Hatch Assay (EHA) and Larval Development Assay (LDA) for albendazole (ABZ), mebendazole (MBZ), fenbendazole (FBZ), thiabendazole (TBZ) and ivermectin (IVM) (only for LDA). FECRT showed that intestinal nematodes developed resistance phenomena against the ABZ-based product, with an extensivity of 80% at seven days post therapy, an egg reduction percentage of 41.89% at seven days post-therapy and 43.9% at 14 days post-therapy. The in vitro EHA highlighted a superior efficacy of TBZ (egg hatch percentage at reference concentration being 51.21) compared to ABZ (71.89%), MBZ (84.46%) and FBZ (79.22%), with a minimum risk of anthelmintic resistance. The LDA test revealed the superior efficacy of FBZ (MIC 0.59 mcg/ml) and IVM (MIC 0.078 mcg/ml), with a minimal risk of inducing parasitic resistance. All in vivo and in vitro tests revealed a limited ABZ efficacy, recommending avoiding the therapy with this substance.

scholarly journals In vitro anthelmintic efficacy of Citrullus colocynthis (L.) Schrad on Haemonchus contortus

2021 ◽  
Vol 91 (3) ◽  
pp. 309-318
Author(s):  
Tauseef Rehman ◽  
◽  
Khalid J. Iqbal ◽  
Azra Anwer ◽  
Rao Z. Abbas ◽  
...  
Keyword(s):  
Ethyl Acetate ◽  
In Vivo Studies ◽  
Citrullus Colocynthis ◽  
Aqueous Methanol ◽  
Egg Hatch ◽  
Promising Alternative ◽  
Fruit Extracts ◽  
Anthelmintic Efficacy ◽  
Serial Dilutions

Ethno-veterinary medicinal studies associated with traditional uses of the flora of the Cholistan desert have shown that fruits of Citrullus colocynthis are used for the treatment of helminth infections. The present research was designed to evaluate the anthelmintic efficacy of C. colocynthis against H. contortus. The in vitro anthelmintic effects of aqueous-methanol and ethyl acetate fruit extracts of C. colocynthis against H. contortus were determined through egg hatch and adult motility assays. The effect of four serial dilutions of 25 mg/mL of each extract compared to levamisol (0.55 mg/mL) and oxfendazole (three serial dilutions of 25 µg/mL) were studied. Both ethyl acetate and aqueous-methanol extracts paralyzed all adult worms 4h and 8h post-exposure at a dose of 25 mg/mL each. In the egg hatch assay, about 83.67% and 80.67% of H. contortus eggs failed to hatch with the same dose (i.e. 25 mg/mL) of ethyl acetate and CAME extracts, respectively. The results of the present study strongly support fruit extracts of C. colocynthis as a promising alternative to synthetic drugs against H. contortus. These findings will lead to further in vivo studies to investigate the bio-availability of the active ingredients of the plant and the minimum non-lethal concentration required for treatment of haemonchosis in livestock. The anthelmintic effects of C. colocynthis might be attributed to the presence of phenolic acids.

scholarly journals In vitro Efficacy of Albendazole against Strongyle eggs recovered from Trade Goats slaughtered at the Nsukka Abattoir: A Preliminary Survey of Resistance to Albendazole

2021 ◽  
Vol 40 (4) ◽  
pp. 287-295
Author(s):  
I.K. Idika ◽  
V.J. Ebuk ◽  
E.I. Okoro ◽  
T.A. Nzeakor ◽  
N.M. Uzonnah ◽  
...  
Keyword(s):  
Faecal Sample ◽  
Anthelmintic Resistance ◽  
Egg Hatch ◽  
Lc50 Value ◽  
Nematode Parasites ◽  
Faecal Culture ◽  
World Association ◽  
Egg Hatch Assay ◽  
Set Up

The efficacy of Albendazole against trichostrongyle nematode parasites in goats presented for slaughter at the Nsukka municipal  abattoir was evaluated using the In vitro Egg hatch assay (EHA) model. The abattoir was visited once every week for 4 consecutive months during which a total of 240 goats were sampled. Fecal samples were collected per rectum from a minimum of 15 goats on each day of the visit. Egg Hatch Assay was performed on strongyle eggs recovered from pooled faecal sample on each day of sampling with a 2.5% W/V Albendazole. Faecal culture was also set up from the pooled faecal sample on each sampling day to recover and identify the nematode parasites present in the goats. Among the 240 goats sampled, the prevalence of trichostrongylosis as observed by the presence of strongyle eggs was 94.6% (227/240). Faecal culture and larval identification revealed 69.8% of the strongyles as  Haemonchus contortus, while 25.5 and 4.8% were Trichostrongylus colubriformis and Oesophagostomum species respectively. In the EHA, Albendazole had mean LC50 value of 0.16 µg/ml which is slightly in excess of the discriminating dose of 0.1µg/ml as prescribed by the World Association for the Advancement of Veterinary Parasitology (WAAVP) as an indication of anthelmintic resistance. There is therefore an urgent need to screen the nematode parasite population in the Nigeria for the presence Albendazole resistance genes. Key words: GI nematode; egg hatch assay; goat; Albendazole; Resistance; Nigeria

scholarly journals In Vitro Anthelmintic Activity of Crude Extracts of Artemisia herba-alba and Punica granatum against Haemonchus contortus

2020 ◽  
Vol 2020 ◽  
pp. 1-7 ◽  
Author(s):  
Aliyi Hassen Ahmed ◽  
Mebrat Ejo ◽  
Teka Feyera ◽  
Dereje Regassa ◽  
Bahar Mummed ◽  
...  
Keyword(s):  
Haemonchus Contortus ◽  
Anthelmintic Activity ◽  
Punica Granatum ◽  
Limiting Factor ◽  
Egg Hatching ◽  
Traditional Use ◽  
Egg Hatch ◽  
Inhibition Effect ◽  
Crude Extracts

Gastrointestinal nematodes (GINs) are the major limiting factor for the successfulness of livestock production throughout the world. Emergence of resistance strains as well as scarcity and high cost of the currently available drugs has led to the evaluation of other alternative helminth control options, mainly from plants. The current study is aimed at investigating the in vitro anthelmintic efficacy of crude methanolic extracts of two traditionally important medicinal plants, Artemisia herba-alba and Punica granatum, against Haemonchus contortus using adult motility assay (AMA) and egg hatch inhibition assay (EHIA). Four graded concentrations of the extracts were tested for both the AMA (10, 5, 2.5, and 1.25 mg/mg) and EHIA (0.1, 0.25, 0.5, and 1 mg/mL) in replicates. Albendazole and phosphate-buffered saline (AMA) or distilled water (EHIA) were used as the positive and negative controls, respectively. The crude extracts of A. herba-alba and P. granatum exhibited a potential anthelmintic activity at all dose levels in a concentration- and time-dependent fashion. The highest concentration (10 mg/mL) of all the extracts caused a significantly (p<0.05) superior nematocidal activity compared to the negative control. Moreover, significant and concentration-dependent egg hatching inhibition effect was observed from both plant extracts. Maximal (98.67%) egg hatching inhibition effect was exhibited by the flower extract of A. herba-alba at 1 mg/mL concentration. The relative egg hatch inhibition efficacy indicated that both plants caused a significantly (p<0.05) greater egg hatch inhibition within 48 hr of exposure. The current study validated the traditional use of both plants as a natural anthelmintic against H. contortus justifying a need to undertake detail pharmacological and toxicological investigation on both plants.

Composition and Potential Health Benefits of Pomegranate: A Review

2019 ◽  
Vol 25 (16) ◽  
pp. 1817-1827 ◽  
Author(s):  
Vesna Vučić ◽  
Milkica Grabež ◽  
Armen Trchounian ◽  
Aleksandra Arsić
Keyword(s):  
Current Knowledge ◽  
Health Benefits ◽  
Punica Granatum ◽  
In Vivo Studies ◽  
Full Potential ◽  
Fruit Peel ◽  
Potential Health ◽  
Comprehensive Search

Background:: Pomegranate (Punica granatum L.) fruits are widely consumed and used as preventive and therapeutic agents since ancient times. Pomegranate is a rich source of a variety of phytochemicals, which are responsible for its strong antioxidative and anti-inflammatory potential. Objective:: The aim of this review is to provide an up-to-date overview of the current knowledge of chemical structure and potential health benefits of pomegranate. Method: : A comprehensive search of available literature. Results:: The review of the literature confirms that juice and extracts obtained from different parts of this plant, including fruit peel, seeds, and leaves exert health benefits in both in vitro and in vivo studies. The antidiabetic, antihypertensive, antimicrobial and anti-tumour effects of pomegranate fruit are of particular scientific and clinical interest. Conclusion:: Further investigations are required to clarify the mechanism of action of the bioactive ingredients and to reveal full potential of pomegranate as both preventive and therapeutic agent.

Supremacy of Synergism: A comparison of anticancer activity of Rhizome Extract of Bistorta amplexicualis and gallic acid in Cancer Cell Lines and Primary Cells

2020 ◽  
Vol 01 ◽  
Author(s):  
Salma Batool ◽  
M. Javaid Asad ◽  
Muhammad Arshad ◽  
Rahman Shah Zaib Saleem ◽  
Muhammad Sheeraz Ahmad
Keyword(s):  
Gallic Acid ◽  
Cell Line ◽  
Cancer Cell ◽  
Plant Extract ◽  
Umbilical Vein ◽  
Cancer Cell Line ◽  
In Vivo Studies ◽  
Therapeutic Window

Background: Bistorta amplexicaulis is a seasonal herb with several folkloric uses. The plant extract has been shown to possess various activities including antioxidant, anticancer, anti-bacterial, anti-fungal, cardio-protective, anti-atherosclerosis activities. Objective: The aim of the study was to quantify the activity of the plant extract and relate it to the activity of the isolated compounds, gallic acid. Methods: Extraction of the plant was carried out. Then the activity of the extract was compared with its constituent, gallic acid. Cytotoxic potential of the two against human liver cancer cell line (HepG2), breast cancer cell line (MCF-7) and human umbilical vein endothelial cells (HUVEC) was evaluated through MTS assay. Results: The extract had better activity against HepG2 as compared to gallic acid (IC50 29µg/mL vs 37µg/mL). It also provided a better therapeutic window by having lower toxicity for HUVEC cells than gallic acid (IC50 63µg/mL vs 47µg/mL) suggesting the use of the extract over the purified gallic acid for these cells. We also performed the fluorescence study of the rhizome extract in ethanol (REE), methanol (REM), 80% ethanol (80RE), 80% methanol, (80RM) and acetone (RAC). The highest intensity of fluorescence was found in REE with excitation at 394 nm and emission at 421nm. Conclusion: The comparison of gallic acid with ethanolic rhizome extract of B. amplexicaulis reveals important insights about utilizing the plant extract over purified gallic acid. The ethanolic extract also has the potential to be used as autoflouresent drug during in vitro and in vivo studies.

scholarly journals Does the in vitro egg hatch test predict the failure of benzimidazole treatment in Haemonchus contortus?

Parasite ◽  
2021 ◽  
Vol 28 ◽  
pp. 62
Author(s):  
Michal Babják ◽  
Alžbeta Königová ◽  
Michaela Urda Dolinská ◽  
Tomas Kupčinskas ◽  
Jaroslav Vadlejch ◽  
...  
Keyword(s):  
Haemonchus Contortus ◽  
Gastrointestinal Parasites ◽  
Resistant Isolate ◽  
Egg Hatch ◽  
Larval Stages ◽  
Tubulin Isotype ◽  
Faecal Egg Count ◽  
Egg Hatch Test

Considerable research has been directed towards optimising in vitro tests that can diagnose resistance in pre-parasitic stages of parasites. The objective of this study was to compare the in vivo faecal egg count reduction test (FECRT), the in vitro egg hatch test (EHT), and the molecular determination of the frequency of a codon 200 allele of β-tubulin isotype 1 associated with benzimidazole resistance in larval stages of Haemonchus contortus obtained from infected goats. Animals were infected with composite infective doses representing 10, 20, 30, 40, 60, and 80% resistant alleles. Faecal samples for the EHT were collected on 28, 33, and 35 days post-infection. The results of the in vivo FECRT indicated that albendazole treatment reduced infections consisting of composite doses of 10, 20, 30, 40, 60, and 80% larvae of the resistant isolate by 91.3, 78.0, 63.3, 48.4, 36.5, and 41.4%, respectively. The drug concentration at which 50% of the eggs were prevented from developing hatching larvae (ED50) in the in vitro EHT varied from 0.09 ± 0.01 to 15.63 ± 12.10 μg/mL thiabendazole. The results of the in vitro EHT indicated that the test could estimate in vivo resistance well. The EHT could thus accurately estimate the in vivo efficacy of the drug and percentage of the resistance allele in the population using hatching parameters in delineation doses. This finding was also supported by comparing the FECRT data to the hatching percentages in the EHT on 30 goat farms in Slovakia with natural mixed infections of gastrointestinal parasites.

scholarly journals Oxfendazole Resistance in Gastrointestinal Nematodes of Beetal Goats at Livestock Farms of Punjab (Pakistan)

2007 ◽  
Vol 76 (1) ◽  
pp. 79-85 ◽  
Author(s):  
M. Saeed ◽  
Z. Iqbal ◽  
A. Jabbar
Keyword(s):  
Anthelmintic Resistance ◽  
Gastrointestinal Nematodes ◽  
Post Treatment ◽  
Egg Hatch ◽  
Treatment Frequency ◽  
Contributory Factors ◽  
Reduction Test ◽  
Egg Hatch Assay

This study was carried out to screen goat farms for anthelmintic resistance (AR) against oxfendazole (OXF) and to determine contributory factors for its development. For this purpose, Beetal goat farms (n = 18) were randomly selected, with natural mixed gastrointestinal nematodosis infection. In vivo (faecal egg count reduction test) and in vitro (egg hatch assay) tests were used to ascertain the presence of AR while a scorecard was used to determine the role of possible contributory factors for oxfendazole resistance. For in vivo test, the experimental animals were divided into two groups of 10 animals each; one group received OXF treatment, while the other served as control. Pre- and post-treatment coproculture was performed to identify the species and genera of nematodes. Egg hatch assay (EHA) was used to confirm the results of FECRT. Fecal egg count reduction test (FECRT) revealed the development of resistance on six farms and post-treatment larval cultures indicated Haemonchus contortus, Trichostrongylus colubriformis, Cooperia curticei, Teladorsagia circumcincta and Oesophagostomum spp. as dominant species with resistance. Furthermore, EHA confirmed the results of FECRT. Among the presumptive factors for AR, the highest composite score was for rotation of anthelmintics followed by treatment frequency, dose rate and nature of medication. The scorecard for the development of AR, used in this study, may be helpful for the assessment of contributory factors of AR.

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