scholarly journals Effect of in vitro aqueous extract of fresh cassava leaves (Manihot esculenta, Crantz) on the larvae of Haemonchus contortus as intestinal parasite of sheep

2021 ◽  
Author(s):  
Ana Flávia Lopes Medeiros ◽  
Marney Pascoli Cereda Cereda ◽  
Gabriel Furrer Matos Braz
Keyword(s):  
Phenolic Compounds ◽  
Aqueous Extract ◽  
Haemonchus Contortus ◽  
Water Extract ◽  
Live Weight ◽  
Manihot Esculenta Crantz ◽  
Cassava Leaves ◽  
Cassava Plant

Abstract Animals feeding with cassava is frequent. In the case of sheep, the producer relates consumption to a reduction in the parasite load. The literature has proven the effect of phenolic compounds as an anthelmintic in vivo, but no evidence for cyanogenic compounds, also present in all parts of the cassava plant, was found. A controlled in vitro bioassay was used to evaluate the aqueous extract of fresh cassava leaves. The efficiency parameter was the imobility of Haemonchus contortus larvae at stage L3, also used to evaluate commercial anthelmintics. Culture plates with 100 active L3 larvae per well were used, constituted a replicate of a total of three. It was placed in these wells, water extract of fresh cassava leaves (FCL) macerated in water, Ivomec® 0.01% (PCI) as positive control and distilled water as negative one (NCW). By considering the immobility of the larvae as a positive anthelmintic effect, the results showed that in NCW all larvae were mobile, while in PCI all 300 larvae were immobile. FCL produced a gradient of larval inactivation correlation (r2 0.996). The best fit equation was y = -33.39ln(x) + 40.517, a logarithmic one, which allowed to calculate the Lethal Concentration (CL) of 3. 44µg.CN-.ml, or 80.0 mg of fresh cassava leaves per milliliter, with performance equivalent to ivermectin. This concentration of free cyanide is compatible with safe consumption of fresh leaves by live weight of sheep. The exact amount of cassava roots, leaves or shoots consumed to provide an effective dose for controlling Haemonchus contort should be established in vivo. Although phenolic compounds must also be present in the extract, immobility was attributed to cyanogenic compounds since the correlation was proportional to the increase in cyanide concentration. It can be concluded that the consumption of fresh cassava leaves has potential as an anthelmintic agent to be evaluated in vivo by feeding sheep and goats. Local use could also add value to the production of fresh cassava leaves, with an average potential production of 2.5 t/ha, available throughout the year, with greater production at the beginning of cultivation and in the summer months. These leaves could be used after the roots harvesting or even in pruning for this purpose. Currently, this volume of good quality protein material is in the field, unused.

scholarly journals In vitro effect of aqueous extract of fresh cassava leaves (Manihot esculenta, Crantz) on the larvae of Haemonchus contortus as an intestinal parasite of sheep.

2021 ◽  
Author(s):  
Ana Flávia Lopes Medeiros ◽  
Marney Pascoli Cereda Cereda ◽  
Gabriel Furrer Matos Braz
Keyword(s):  
Phenolic Compounds ◽  
Aqueous Extract ◽  
Haemonchus Contortus ◽  
Live Weight ◽  
Manihot Esculenta Crantz ◽  
Cassava Leaves ◽  
Cassava Plant ◽  
Vitro Effect

Abstract Cassava is frequently fed to animals. In the case of sheep, the producer relates consumption to a reduction in the parasite load. The literature has proven the effect of phenolic compounds as an anthelmintic in vivo, but no evidence for cyanogenic compounds, also present in all parts of the cassava plant, was found. A controlled in vitro bioassay was used to evaluate the aqueous extract of fresh cassava leaves. The efficiency parameter was the immobility of Haemonchus contortus larvae at the L3 stage, also used to evaluate commercial anthelmintics. Cell culture plates with 100 active L3-stage larvae per well were used, being each replicate constituted by three wells. Aqueous extract of fresh cassava leaves (FCL), Ivomec® 0.01% (PCI) as the positive control, and distilled water as negative control (NCW), were placed in the culture plate wells. Considering the immobility of the larvae as a positive anthelmintic effect, the results showed that in NCW treatments all larvae were mobile, while in PCI all 300 larvae were immobile. FCL produced a gradient of larval inactivation correlation (R2 0.996). The best-fit equation was y = -33.39ln(x) + 40.517, a logarithmic equation, which allowed the calculation of the Lethal Concentration (CL) of 3.44 µg CN- ml, or 80.0 mg of fresh cassava leaves per milliliter of water, with a performance equivalent to ivermectin. This concentration of free cyanide is compatible with the safe consumption of fresh leaves by live weight of sheep. The exact amount of cassava roots, leaves, or shoots, consumed to provide an effective dose for controlling H. contort should be established in vivo. Although phenolic compounds must also be present in the extract, the immobility was attributed to cyanogenic compounds since the correlation was proportional to the increase in the cyanide concentration. It can be concluded that the consumption of fresh cassava leaves has the potential as an anthelmintic agent to be evaluated in vivo by feeding sheep and goats. Local use could also add value to the production of fresh cassava leaves, with an average potential production of 2.5 tonnes ha-1, available throughout the year, with greater production at the beginning of cultivation and in the summer months. These leaves could be used after the roots harvesting or even after pruning for this purpose. Currently, this amount of good quality protein material remains without use in the field.

scholarly journals In vitro action of Mimosa caesalpinifolia ketone extract on Haemonchus contortus and Trichostrongylus colubriformis

2017 ◽  
Vol 38 (4) ◽  
pp. 1963 ◽  
Author(s):  
Danilo Rodrigues Barros Brito ◽  
Livio Martins Costa Júnior ◽  
João Luis Garcia ◽  
Suzana Gomes Lopes ◽  
Giselle Cutrim de Oliveira Santos ◽  
...  
Keyword(s):  
Phenolic Compounds ◽  
Haemonchus Contortus ◽  
Anthelmintic Activity ◽  
In Vivo Studies ◽  
In Vitro Assays ◽  
Trichostrongylus Colubriformis ◽  
Lower Efficiency ◽  
Egg Hatchability

This study aimed to evaluate the in vitro anthelmintic activity of Mimosa caesalpinifolia (sabiá) ketone extract, which is rich in condensed tannins (CT), on Haemonchus contortus and Trichostrongylus colubriformis. The leaves and stems of M. caesalpinifolia were collected, dried in the shade, and ground to a dry powder from which ketone extracts were obtained for in vitro assays. The extracts were diluted in 5% acetone and PBS to concentrations in the range 0.25-2.80 mg mL-¹ and the inhibition of egg hatchability was assayed. Feces collected from goats naturally infected with H. contortus (55%) and T. colubriformis (45%) were macerated and the helminth eggs were collected to conduct in vitro egg hatchability tests. Nematode larvae collected from the feces were used for a larval exsheathment inhibition assay of M. caesalpinifolia leaf and M. caesalpinifolia leaf and stem ketone extracts at concentrations of 0.6 and 1.2 mg mL-¹; three replicates were performed for each concentration. PBS was used as a negative control. Polyvinylpolypyrrolidone (PVPP) was added to the extracts to assess the effect of phenolic compounds on anthelmintic activity. An inhibitory effect of 80.7% and 82.3% was observed on hatchability after treatment with 1.0 and 2.0 mg mL-¹ M. caesalpinifolia leaf ketone extract, respectively. An inhibition rate lower than 75% was observed for 1.0 mg mL-¹ M. caesalpinifolia leaf and stem ketone extract. The M. caesalpinifolia leaf ketone extract larval exsheathment assay resulted in inhibition of 97.3% and 99.8% at concentrations of 0.6 and 1.2 mg mL-¹, respectively. For the leaf and stem ketone extract, the larval exsheathment inhibition was 94.3% at 1.2 mg mL-¹. The leaf and leaf and stem extracts showed no inhibitory effects on larval exsheathment after the addition of PVPP. These results showed that cuticular loss was related to the action of the phenolic compounds in the M. caesalpinifolia extracts. M. caesalpinifolia leaf ketone extract inhibited egg hatchability, but the leaf and stem extract showed lower efficiency, which indicated a greater abundance of active compounds in the leaves. The M. caesalpinifolia ketone extracts were effective in the inhibition of larval exsheathment at the highest concentration tested. Based on these results, in vivo studies should be performed to demonstrate the anthelmintic activity of this plant in goats.

scholarly journals HPLC ANALYSIS AND ANTI-INFLAMMATORY PROPERTIES STUDIES OF TRUNK BARKS OF ACACIA NILOTICA VAR ADANSONII (GUILL AND PERR) O KTZE (MIMOSACEAE)

2021 ◽  
pp. 40-46
Author(s):  
BOLY A. GILCHRIST L. ◽  
OUEDRAOGO NOUFOU ◽  
TRAORE ARISTIDE ◽  
TRAORE KADIATOU T. ◽  
BELEMLILGA MOHAMED ◽  
...  
Keyword(s):  
Acetic Acid ◽  
Phenolic Compounds ◽  
Ethyl Acetate ◽  
Aqueous Extract ◽  
Hplc Analysis ◽  
Acacia Nilotica ◽  
Anti Inflammatory ◽  
Different Doses

Objective: The objective of this study was to evaluate the in vitro and in vivo anti-inflammatory properties of the aqueous extract and fractions of the trunk bark of Acacia nilotica. Methods: A maceration of the powder of the trunks barks of the plant was realized. Then the aqueous macerate obtained was fractionated with dichloromethane, butanol and ethyl acetate successively. The phenolic compounds of the aqueous extract, butanol and ethyl acetate fractions were identified by HPLC/DAD. Lipoxygenase and phospholipase inhibition tests with the aqueous extract and the butanol and ethyl acetate fractions were carried out. The anti-inflammatory potential of the aqueous extract was assessed in vivo by the anti-edema test with carrageenan and the analgesic test with acetic acid at different doses (200 mg/ml; 400 mg/ml; 600 mg/ml). Aspirin (200 mg/ml) and paracetamol (200 mg/ml) were used as a reference. Results: The HPLC/DAD analysis of the extracts revealed that gallic acid is the most abundant phenol acid in the extracts. The aqueous extract inhibited lipoxygenase (IC50 = 18.32±1.18 μg/ml), phospholipase (11.44±0.32% per 100 μg/ml) and cyclooxygenase (56.48±0.29% for 100 μg/ml) as well as its tested fractions. It also reduced edema and pain in the mice by more than 50% from the 400 mg/ml dose. Conclusion: Aqueous extract of Acacia nilotica has anti-inflammatory properties. Hence its use in traditional medicine in the treatment of inflammation.

scholarly journals Anthelmintic effect of Indigofera tinctoria L on Haemonchus contortus obtained from sheep in Indonesia

2021 ◽  
pp. 1272-1278
Author(s):  
Iskandar Muda ◽  
Joko Prastowo ◽  
Wisnu Nurcahyo ◽  
Sarmin Sarmin
Keyword(s):  
Weight Gain ◽  
Aqueous Extract ◽  
Haemonchus Contortus ◽  
Saline Water ◽  
Positive Control ◽  
Negative Control ◽  
Indigofera Tinctoria ◽  
Anthelmintic Effect

Background and Aim: Hemonchosis has resulted in huge economic losses for sheep farmers worldwide. Secondary metabolite compounds from Indigofera tinctoria L. can be used as anthelmintics. This study aimed to evaluate the in vitro and in vivo effects of I. tinctoria L. aqueous extract (IAE) as an anthelmintic against adult Haemonchus contortus isolated from sheep. Materials and Methods: Ten active adult worms were placed in each Petri dish containing 25 mL of IAE, each having a different concentration of IAE (at concentrations 100, 120, 140, 160, 180, 200, and 220 mg/mL). Each experiment was repeated. The positive control used 1% albendazole, and the negative control used 0.62% saline water. The number of immobile worms and the time of mortality were recorded after 2, 4, 6, and 8 h. The dead worms were subsequently tested using scanning electron microscopy (SEM) and sodium dodecyl sulfate-polyacrylamide gel electrophoresis. In the in vivo study, 15 sheep with an average fecal egg count (FEC) of 1000 eggs per gram were treated with IAE for 9 days: Group A (negative control, saline water 0.62%), Group B (21 mg/mL), Group C (41 mg/mL), Group D (62 mg/mL), and Group E (positive control, albendazole 100 mg/mL). Measurements of the body weight, FEC reduction test, and hematology testing were performed on days 0, 7, and 14. SEM was performed using worms found from the abomasum of slaughtered sheep. Results: The leaves of I. tinctoria L. contained a number of secondary metabolites, including total tannins, saponins, flavonoids, and alkaloids. The most effective concentration that killed the adult H. contortus worms was 220 mg/mL (93.33% mortality) after 8 h of treatment. The electrophoresis results showed that the protein band at a dose of 22% was less than that of the control. The highest FECR value of the treatment group on the 14th day after treatment was at a dose of 62 mg/mL. The highest weight gain as well as the highest increased hemoglobin (Hb), packed cell volume (PCV), and total erythrocyte count (TEC) values on the 14th day after treatment were at a dose of 41 mg/mL. The SEM results showed that IAE treatment caused the worms' anterior parts to become wrinkled with thick creases and cuticle abrasion (in vitro) and the anterior part to shrink along with the presence of aggregates in the worm cuticle (in vivo). Conclusion: The aqueous extract of I. tinctoria contains tannins, saponins, flavonoids, and alkaloids and has an anthelmintic effect with decreased FEC, increased weight gain, Hb, PCV, and TEC, causing damage to the worms' body and reducing the protein profile of adult H. contortus worms.

Inhibitory effects of epimedium herb water extract on the inflammatory response in vitro and in vivo

Planta Medica ◽  
2016 ◽  
Vol 81 (S 01) ◽  
pp. S1-S381
Author(s):  
YC Oh ◽  
YH Jeong ◽  
WK Cho ◽  
SJ Lee ◽  
JY Ma
Keyword(s):  
Inflammatory Response ◽  
Water Extract ◽  
Inhibitory Effects

In vitro and in vivo Trypanocidal Effect of the Aqueous extract of the Leaves of Ochna Schweinfurthiana

2020 ◽  
Vol 4 (1) ◽  
pp. 47-51
Author(s):  
Eteme Enama S ◽  
Messi A N ◽  
Mahob R J ◽  
Siama A ◽  
Njan Nloga A M
Keyword(s):  
Aqueous Extract

In-vitro and in-vivo evaluation of antidiabetic activity of Withania coagulans extract

2019 ◽  
Vol 09 ◽  
Author(s):  
Tejas Patel ◽  
B.N. Suhagia
Keyword(s):  
Blood Glucose ◽  
Acute Toxicity ◽  
Aqueous Extract ◽  
Pancreatic Beta Cell ◽  
Toxicity Study ◽  
Acute Toxicity Study ◽  
Withania Coagulans ◽  
Study Results

Background: Diabetes mellitus is major issue to public health as its prevalence is rising day by day. Synthetic agents available for the diabetic treatment are expensive or produce undesirable side effect on chronic use and some of them are not suitable during pregnancy. Herbal medicines accepted widely due to side effects and low cost. Objective: The aim of present study was to evaluate the activity of Withania coagulans extract using In-vitro and In-vivo model. Methods: Different three types of Withania coagulans extract were prepared using aqueous (W1), Alcohol (W2) and hydro-alcoholic (50:50) mixture (W3). In-vitro Anti-diabetic activity of the all three extracts evaluated using RINm5F Pancreatic beta cells.Further, n-vivo anti-diabetic evaluation performed by administering 50 mg/kg (p.o) aqueous extract for 7 days in Streptozotocin (STZ)-induced mice. Body weight of the animals was also determined to perform acute toxicity study. Results: The results of in –vitro cell based study indicated that among all three extract, aqueous extract (W1) of Withania coagulans showed potential increase in inulin release. The EC50 of the W1 (249.6 µg/L) which is compared with standard (Glibenclamide) EC50. From the results of In-vitro study, W1 subjected for acute toxicity study and the acute toxicity study results indicated LD50 of 50mg/kg. Diabetic rats treated with W1 extract at oral dose of 50 mg/kg for 7 days showed 34.17% reduction in blood glucose in comparison to untreated diabetic (STZ-induced) rats. Blood glucose levels of Standard treated (Glibenclamide) and control untreated. Conclusion: In conclusion, results of pancreatic beta cell based study showed increase in insulin release by administration of extract. Further aqueous extract (W1) was potentially reduced blood glucose level in STZ induced diabetic mice.

scholarly journals Oriental Hornet (Vespa orientalis) Larval Extracts Induce Antiproliferative, Antioxidant, Anti-Inflammatory, and Anti-Migratory Effects on MCF7 Cells

Molecules ◽  
2021 ◽  
Vol 26 (11) ◽  
pp. 3303
Author(s):  
Amina M. G. Zedan ◽  
Mohamed I. Sakran ◽  
Omar Bahattab ◽  
Yousef M. Hawsawi ◽  
Osama Al-Amer ◽  
...  
Keyword(s):  
Phenolic Compounds ◽  
Aqueous Extract ◽  
Antioxidant Activities ◽  
Individual Treatment ◽  
Alcoholic Extract ◽  
Anticancer Effect ◽  
Mcf7 Cells ◽  
Vespa Orientalis ◽  
Anti Inflammatory

The use of insects as a feasible and useful natural product resource is a novel and promising option in alternative medicine. Several components from insects and their larvae have been found to inhibit molecular pathways in different stages of cancer. This study aimed to analyze the effect of aqueous and alcoholic extracts of Vespa orientalis larvae on breast cancer MCF7 cells and investigate the underlying mechanisms. Our results showed that individual treatment with 5% aqueous or alcoholic larval extract inhibited MCF7 proliferation but had no cytotoxic effect on normal Vero cells. The anticancer effect was mediated through (1) induction of apoptosis, as indicated by increased expression of apoptotic genes (Bax, caspase3, and p53) and decreased expression of the anti-apoptotic gene Bcl2; (2) suppression of intracellular reactive oxygen species; (3) elevation of antioxidant enzymes (CAT, SOD, and GPx) and upregulation of the antioxidant regulator Nrf2 and its downstream target HO-1; (4) inhibition of migration as revealed by in vitro wound healing assay and downregulation of the migration-related gene MMP9 and upregulation of the anti-migratory gene TIMP1; and (5) downregulation of inflammation-related genes (NFκB and IL8). The aqueous extract exhibited the best anticancer effect with higher antioxidant activities but lower anti-inflammatory properties than the alcoholic extract. HPLC analysis revealed the presence of several flavonoids and phenolic compounds with highest concentrations for resveratrol and naringenin in aqueous extract and rosmarinic acid in alcoholic extract. This is the first report to explain the intracellular pathway by which flavonoids and phenolic compounds-rich extracts of Vespa orientalis larvae could induce MCF7 cell viability loss through the initiation of apoptosis, activation of antioxidants, and inhibition of migration and inflammation. Therefore, these extracts could be used as adjuvants for anticancer drugs and as antioxidant and anti-inflammatory agents.

scholarly journals High-Throughput Identification of Organic Compounds from Polygoni Multiflori Radix Praeparata (Zhiheshouwu) by UHPLC-Q-Exactive Orbitrap-MS

Molecules ◽  
2021 ◽  
Vol 26 (13) ◽  
pp. 3977
Author(s):  
Shaoyun Wang ◽  
Xiaozhu Sun ◽  
Shuo An ◽  
Fang Sang ◽  
Yunli Zhao ◽  
...  
Keyword(s):  
High Performance ◽  
Chemical Constituents ◽  
Water Extract ◽  
Ethanol Extract ◽  
In Vivo Studies ◽  
Polygonum Multiflorum ◽  
Q Exactive ◽  
Orbitrap Ms

Polygoni Multiflori Radix Praeparata (PMRP), as the processed product of tuberous roots of Polygonum multiflorum Thunb., is one of the most famous traditional Chinese medicines, with a long history. However, in recent years, liver adverse reactions linked to PMRP have been frequently reported. Our work attempted to investigate the chemical constituents of PMRP for clinical research and safe medication. In this study, an effective and rapid method was established to separate and characterize the constituents in PMRP by combining ultra-high performance liquid chromatography with hybrid quadrupole-orbitrap mass spectrometry (UHPLC-Q-Exactive Orbitrap-MS). Based on the accurate mass measurements for molecular and characteristic fragment ions, a total of 103 compounds, including 24 anthraquinones, 21 stilbenes, 15 phenolic acids, 14 flavones, and 29 other compounds were identified or tentatively characterized. Forty-eight compounds were tentatively characterized from PMRP for the first time, and their fragmentation behaviors were summarized. There were 101 components in PMRP ethanol extract (PMRPE) and 91 components in PMRP water extract (PMRPW). Simultaneously, the peak areas of several potential xenobiotic components were compared in the detection, which showed that PMRPE has a higher content of anthraquinones and stilbenes. The obtained results can be used in pharmacological and toxicological research and provided useful information for further in vitro and in vivo studies.

scholarly journals Mutagenicity evaluation of Anastatica hierochuntica L. aqueous extract in vitro and in vivo

2017 ◽  
Vol 243 (4) ◽  
pp. 375-385 ◽  
Author(s):  
Siti Rosmani Md Zin ◽  
Zahurin Mohamed ◽  
Mohammed A Alshawsh ◽  
Won F Wong ◽  
Normadiah M Kassim
Keyword(s):  
Aqueous Extract ◽  
Positive Control ◽  
In Vitro Assay ◽  
In Vivo Study ◽  
Sufficient Evidence ◽  
Aqueous Extracts ◽  
Mutagenic Potential ◽  
Vitro Assay

Anastatica hierochuntica L. ( A. hierochuntica), a folk medicinal plant, was evaluated for mutagenic potential via in vitro and in vivo assays. The in vitro assay was conducted according to modified Ames test, while the in vivo study was performed according to Organisation for Economic Co-operation and Development guideline for mammalian erythrocyte micronucleus assay. Four groups ( n= 5 males and 5 females per group) Sprague Dawley rats were randomly chosen as the negative control, positive control (received a single intramuscular injection of cyclophosphamide 50 mg/kg), 1000 and, 2000 mg/kg A. hierochuntica aqueous extracts. All groups except the positive control were treated orally for three days. Findings of the in vitro assay showed mutagenic potential of AHAE at 0.04 and 0.2 mg/ml. However, no mutagenic effect was demonstrated in the in vivo study up to 2000 mg/kg. No significant reduction in the polychromatic and normochromatic erythrocytes ratio was noted in any of the groups. Meanwhile, high micronucleated polychromatic erythrocytes frequency was seen in cyclophosphamide-treated group only. These findings could perhaps be due to insufficient dosage of A. hierochuntica aqueous extracts to cause genetic damage on the bone marrow target cells. Further acute and chronic in vivo toxicity studies may be required to draw pertinent conclusion on the safety aspect of A. hierochuntica aqueous extracts consumption. Impact statement In this paper, we report on the mutagenicity evaluation of Anastatica hierochuntica aqueous extract. This is a significant research in view of the popularity of this herb consumption by the people across the globe despite of limited scientific evidence on its toxicity potential. This study is intended to encourage more extensive related research in order to provide sufficient evidence and guidance for determining its safe dosage.

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