In vitro and in vivo anthelmintic activity of crude extracts of Coriandrum sativum against Haemonchus contortus

In vitro anthelmintic activities of crude aqueous and hydro-alcoholic extracts of the seeds of Coriandrum sativum (Apiaceae) were investigated on the egg and adult nematode parasite Haemonchus contortus. The aqueous extract of Coriandrum sativum was also investigated for in vivo anthelmintic activity in sheep infected with Haemonchus contortus. Both extract types of Coriandrum sativum inhibited hatching of eggs completely at a concentration less than 0.5 mg/ml. ED₅₀ of aqueous extract of Coriandrum sativum was 0.12 mg/ml while that of hydro-alcoholic extract was 0.18 mg/ml. There was no statistically significant difference between aqueous and hydro-alcoholic extracts (p > 0.05). The hydro-alcoholic extract showed better in vitro activity against adult parasites than the aqueous one. For the in vivo study, 24 sheep artificially infected with Haemonchus contortus were randomly divided into four groups of six animals each. The first two groups were treated with crude aqueous extract of Coriandrum sativum at 0.45 and 0.9 g/kg dose levels, the third group with albendazole at 3.8 mg/kg and the last group was left untreated. Efficacy was tested by faecal egg count reduction (FECR) and total worm count reduction (TWCR). On day 2 post treatment, significant FECR was detected in groups treated with higher dose of Coriandrum sativum (p < 0.05) and albendazole (p < 0.001). On days 7 and 14 post treatment, significant FECR was not detected for both doses of Coriandrum sativum (p > 0.05). Significant (p < 0.05) TWCR was detected only for higher dose of Coriandrum sativum compared to the untreated group. Reduction in male worms was higher than female worms. Treatment with both doses of Coriandrum sativum did not help the animals improve or maintain their PCV while those treated with albendazole showed significant increase in PCV (p < 0.05).     

The In Vitro and In Vivo Biological Activities of the Leaf of Cape Myrtle,Myrsine africana L.

The cape myrtle, Myrsine africana L., is a widely used medicinal plant, which has not been well investigated. We assessed the in vivo hepatoprotective and in vitro antiproliferative and antioxidant effects of leaf extracts of M. africana chemically profiled using high‐performance liquid chromatography. Three flavonoids were quantified, and gas chromatography–mass spectrometry analysis revealed the presence of common fatty acids. The animal study was conducted on mice treated with CCl4, using three doses each of the methanol and chloroform extract (100, 200 and 300 mg/kg b.w.),with silymarin as a positive control. Hepatoprotective effects were determined by analyzing blood for liver marker and antioxidant enzymes, direct bilirubins and total proteins. The methanol extract (300 mg/kg b.w.) showed the strongest hepatoprotective effects against abnormalities produced by CCl4. The in vivo hepatoprotective effects correlated well with the in vitro antioxidant and antiproliferative activities and with high levels of flavonoids in the extracts. Finally, molecular docking studies of the constituent quercetin were undertaken in silico and several sites of binding to human estrogen receptor (ER) protein, linked with alkaline phosphatase, identified. Copyright © 2017 John Wiley & Sons, Ltd.     

In vitro anthelmintic activity of crude extracts of five medicinal plants against egg-hatching and larval development of Haemonchus contortus

Ethnopharmacological relevance

Senna occidentalis, Leonotis ocymifolia, Leucas martinicensis, Rumex abyssinicus, and Albizia schimperiana are traditionally used for treatment of various ailments including helminth infection in Ethiopia.

Materials and Methods

In vitro egg hatch assay and larval development tests were conducted to determine the possible anthelmintic effects of crude aqueous and hydro-alcoholic extracts of the leaves of Senna occidentalis, aerial parts of Leonotis ocymifolia, Leucas martinicensis, Rumex abyssinicus, and stem bark of Albizia schimperiana on eggs and larvae of Haemonchus contortus.

Results

Both aqueous and hydro-alcoholic extracts of Leucas martinicensis, Leonotis ocymifolia and aqueous extract of Senna occidentalis and Albizia schimperiana induced complete inhibition of egg hatching at concentration less than or equal to 1 mg/ml. Aqueous and hydro-alcoholic extracts of all tested medicinal plants have shown statistically significant and dose dependent egg hatching inhibition. Based on ED₅₀, the most potent extracts were aqueous and hydro-alcoholic extracts of Leucas martinicensis (0.09 mg/ml), aqueous extracts of Rumex abyssinicus (0.11 mg/ml) and Albizia schimperiana (0.11 mg/ml). Most of the tested plant extracts have shown remarkable larval development inhibition. Aqueous extracts of Leonotis ocymifolia, Leucas martinicensis, Albizia schimperiana and Senna occidentalis induced 100, 99.85, 99.31, and 96.36% inhibition of larval development, respectively; while hydro-alcoholic extracts of Albizia schimperiana induced 99.09 inhibition at the highest concentration tested (50 mg/ml). Poor inhibition was recorded for hydro-alcoholic extracts of Senna occidentalis (9%) and Leonotis ocymifolia (37%) at 50 mg/ml.

Conclusions

The overall findings of the current study indicated that the evaluated medicinal plants have potential anthelmintic effect and further in vitro and in vivo evaluation is indispensable to make use of these plants.     

Anthelmintic activity of Chenopodium album (L) and Caesalpinia crista (L) against trichostrongylid nematodes of sheep

AIM OF THE STUDY: The present study was carried out to determine the anthelmintic activity of Caesalpinia crista (L.) (Fabaceae) seed kernel and Chenopodium album (L.) (Chenopodiaceae) whole plant in order to justify their traditional use in veterinary medicine. MATERIALS AND METHODS: In vitro anthelmintic activity of crude aqueous methanolic extract (AME) of both the plants was determined using mature Haemonchus contortus and their eggs in adult motility assay and egg hatch test, respectively. In vivo anthelmintic activity was evaluated in sheep naturally infected with mixed species of gastrointestinal nematodes by administering crude powder (CP) and AME in increasing doses (1.0-3.0 g/kg). RESULTS: Both plants exhibited dose- and time-dependent anthelmintic effects by causing mortality of worms and inhibition of egg hatching. Caesalpinia crista (LC50=0.134 mg/mL) was found to be more potent than Chenopodium album (LC50=0.449 mg/mL) in egg hatch test. In vivo, maximum reduction in eggs per gram (EPG) of faeces was recorded as 93.9 and 82.2% with Caesalpinia crista and Chenopodium album AME at 3.0 g/kg on day 13 and 5 post-treatment, respectively. Levamisole (7.5 mg/kg), a standard anthelmintic agent, showed 95.1-95.6% reduction in EPG. CONCLUSIONS: These data show that both Caesalpinia crista and Chenopodium album possess anthelmintic activity in vitro and in vivo, thus, justifying their use in the traditional medicine system of Pakistan.     

Anthelmintic activity of Arisaema franchetianum and Arisaema lobatum essential oils against Haemonchus contortus

Ethnopharmacological relevance

Arisaema franchetianum and Arisaema lobatum are two perennial plants native to China. Arisaema franchetianum is universally used to promote the subsidence of induration and swelling, quicken blood and relieve pains, and kill intestinal parasites in humans and animals. Arisaema lobatum is used to treat malaria, intestinal parasites, and snake and insect bites in humans and animals. The aim of this study was to determine the composition of the essential oils from Arisaema franchetianum and Arisaema lobatum and evaluate the anthelmintic effect against Haemonchus contortus.

Materials and methods

Two oils were investigated by GC and GC–MS. The anthelmintic bioassay tests of Arisaema franchetianum and Arisaema lobatum essential oil, linalool and carvacrol were performed using egg hatch assay (EHA), larval development assay (LDA) and larval migration inhibition assay (LMIA).

Results

Fifty six components representing 96.88% of the Arisaema franchetianum oil and 64 components representing 96.88% of the Arisaema lobatum oil were identified. Carvacrol and linalool were found to be the major constituents of two oils. In the EHA, greater than 99% inhibition were observed with Arisaema franchetianum oil at 10 mg/mL (CE₅₀ 1.63 mg/mL) and Arisaema lobatum oil at 5 and 10 mg/mL (CE₅₀ 0.48 mg/mL). In the LDA, both oils induced complete inhibition at 10 mg/mL, with the CE₅₀ being 1.10 mg/mL for Arisaema franchetianum oil and 0.73 mg/mL for Arisaema lobatum oil. In the LMIA, the Arisaema franchetianum oil and Arisaema lobatum oil at best inhibited 74.1% and 95.6% of larval migration at 10 mg/mL, respectively. Carvacrol exhibited similar activity to Arisaema lobatum essential oil and linalool did not show high activity in every assay.

Conclusions

These data show for the first time that the essential oils obtained from Arisaema franchetianum or Arisaema lobatum had promising anthelmintic activity against Haemonchus contortus. Arisaema plant may offer an alternative source for the control of gastrointestinal nematodes of sheep and goats.     

Anthelmintic activity of Ziziphus nummularia (bark) and Acacia nilotica (fruit) against Trichostrongylid nematodes of sheep

Ethnopharmacological relevance

Ziziphus nummularia (Rhamnaceae) and Acacia nilotica (Fabaceae) are being used as anthelmintics in ethnoveterinary medicinal system of Pakistan.

Aim of the study

Present study was conducted to determine the anthelmintic activity of Ziziphus nummularia (bark) and Acacia nilotica (fruit) in order to justify their traditional use in veterinary medicine.

Materials and methods

In vitro anthelmintic activity of crude methanolic extract (CME) of both the plants was determined against Haemonchus contortus by the adult motility assay, the egg hatch test and the larval development assay. In vivo anthelmintic activity was evaluated in sheep naturally infected with gastrointestinal nematodes by administering increasing doses of crude powder (CP) and CME (1.0–3.0 g/kg).

Results

Both the plants exhibited dose- and time-dependent anthelmintic effects by causing mortality of worms, and inhibiting egg hatching and larval development. Acacia nilotica (LC₅₀ = 512.86 and 194.98 μg/ml) was found to be more potent than Ziziphus nummularia (LC₅₀ = 676.08 and 398.11 μg/ml) in egg hatch test and larval development assay, respectively. In vivo, maximum fecal egg count reduction (84.7%) was recorded on day 13 post-treatment in sheep treated with Ziziphus nummularia CME (3.0 g/kg) followed by 78.5% on same day with Acacia nilotica CME (3.0 g/kg).

Conclusions

These data show that both Ziziphus nummularia and Acacia nilotica possess anthelmintic activity in vitro and in vivo, justifying their use in traditional veterinary medicine in Pakistan.     

Antimalarial activity of methanolic extracts from plants used in Kenyan ethnomedicine and their interactions with chloroquine (CQ) against a CQ-tolerant rodent parasite, in mice

Methanolic extracts from 15 medicinal plants representing 11 families, used traditionally for malaria treatment in Kenya were screened for their in vivo antimalarial activity in mice against a chloroquine (CQ)-tolerant Plasmodium berghei NK65, either alone or in combination with CQ. The plant parts used ranged from leaves (L), stem bark (SB), root bark (RB), seeds (S) and whole plant (W). When used alone, extracts from seven plants, Clerodendrum myricoides (RB), Ficus sur (L/SB/RB), Maytenus acuminata (L/RB), Rhamnus prinoides (L/RB), Rhamnus staddo (RB), Toddalia asiatica (RB) and Vernonia lasiopus (RB) had statistically significant parasitaemia suppressions of 31.7-59.3%. In combination with CQ, methanolic extracts of Albizia gummifera (SB), Ficus sur (RB), Rhamnus prinoides and Rhamnus staddo (L/RB), Caesalpinia volkensii (L), Maytenus senegalensis (L/RB), Withania somnifera (RB), Ekebergia capensis (L/SB), Toddalia asiatica (L/RB) and Vernonia lasiopus (L/SB/RB) gave statistically significant and improved suppressions which ranged from 45.5 to 85.1%. The fact that these activities were up to five-fold higher than that of extract alone may suggest synergistic interactions. Remarkable parasitaemia suppression by the extracts, either alone or in combination with CQ mostly resulted into longer mouse survival relative to the controls, in some cases by a further 2 weeks. Plants, which showed significant antimalarial activity including Vernonia lasiopus, Toddalia asiatica, Ficus sur, Rhamnus prinoides and Rhamnus staddo warrant further evaluation in the search for novel antimalarial agents against drug-resistant malaria.     

Antimicrobial activity of selected plant species from “the Argentine Puna” against sensitive and multi-resistant bacteria

Aim

The plant species reported here are traditionally used in the “Puna” or “Altiplano” of Argentina for ailments related to bacterial infections. The aim of this study was to evaluate their antimicrobial properties against a panel of sensitive and multi-resistant Gram-positive and Gram-negative bacteria.

Materials and methods

The antimicrobial activity of tinctures and aqueous extracts (Baccharis boliviensis, Chiliotrichiopsis keidelii, Chuquiraga atacamensis, Fabiana bryoides, Fabiana densa, Fabiana punensis, Frankenia triandra, Parastrephia lucida, Parastrephia lepidophylla, Parastrephia phyliciformis, Tetraglochin cristatum) was determined using the agar macrodilution and broth microdilution methods recommended by the Clinical and Laboratory Standards Institute (CLSI, formerly NCCLS). The antibiotic resistant clinical strains were isolated from nosocomial infection in human lesions of skin and soft parts.

Results

The ethanolic extracts of 11 plant species inhibited the growth of one or more of the following strains: Staphylococcus aureus, Enterococcus faecalis, Escherichia coli, Klebsiella pneumoniae, Proteus mirabilis, Enterobacter cloacae, Morganella morganii, Pseudomonas aeruginosa. Ethanol extracts (tinctures) of aerial parts of Baccharis, Fabiana and Parastrephia showed the highest levels of antibacterial activity on methicillin, oxacillin and gentamicin resistant Staphylococcus with MIC values from 20 to 150 μg/ml. Baccharis boliviensis and Fabiana bryoides were more active than the other plant species on Enterococcus faecalis with different phenotype. The most interesting activity on multi-resistant Gram-negative strains was obtained from Chuquiraga atacamensis. Parastrephia species showed activity against Enterobacter cloacae, Pseudomonas aeruginosa and Proteus mirabilis. The ethanolic extracts exhibited stronger activity and broader spectrum of action than aqueous extracts. The extracts were bactericidal in most cases.

Conclusions

The presence of antibacterial activity in Puna plant extracts against multi-resistant bacteria give support to their traditional use for treating conditions associated with microorganisms in humans and animals and consequently seems promising for the treatment of multi-resistant bacteria.     

Potential of medicinal plants from the Brazilian semi-arid region (Caatinga) against Staphylococcus epidermidis planktonic and biofilm lifestyles

Ethnopharmacological relevance

Medicinal plants from the Caatinga, a Brazilian xeric shrubland, are used in folk medicine to treat infections. These ethnopharmacological data can contribute to obtaining new antimicrobial/antibiofilm extracts and natural product prototypes for the development of new drugs. The aim of this study was to investigate the antibiofilm and antibacterial activities of 45 aqueous extracts from 24 Caatinga plant species.

Materials and methods

The effect of aqueous extracts on planktonic cells and on biofilm formation by Staphylococcus epidermidis was studied by the OD₆₀₀ absorbance and by the crystal violet assay, respectively. Scanning electron microscopy (SEM) was used to generate comparative images of extract-treated and untreated biofilms. Chromatographic analyses were performed to characterize the active extracts.

Results

The in vitro screening, at 0.4 mg/mL and 4.0 mg/mL, showed 20 plants effective in preventing biofilm formation and 13 plants able to inhibit planktonic bacterial growth. SEM images demonstrated distinct profiles of bacterial adhesion, matrix production and cell morphology according to different treatments and surfaces. The phytochemical analysis of the selected active extracts indicates the polyphenols, coumarins, steroids and terpenes as possible active compounds.

Conclusion

This study describes the first antibiofilm and antibacterial screening of Caatinga plants against S. epidermidis. The evaluation presented in this study confirms several ethnopharmacological reports and can be utilized to identify new antibiofilm and antibacterial products against S. epidermidis from traditional Brazilian medicine.     

Genotoxic and clastogenic activity of saponins extracted from Nauclea bark as assessed by the micronucleus and the comet assays in Chinese Hamster Ovary cells

Ethnopharmacological relevance

Bark extracts of Nauclea latifolia, Nauclea diderrichii, Nauclea pobeguinii and Nauclea vandergutchii are used in traditional medicine in West and South Africa for the treatment of fevers, diarrhea and malaria.

Aim of the study

To estimate the possible long-term toxicity and genotoxicity of plant extracts (dichloromethane, methanol, water/methanol, water) and saponins.

Materials and methods

The clastogenicity of plant extracts and saponins was assessed by the micronucleus assay performed on Chinese Hamster Ovary cells. The DNA-damaging activity of saponin mixture was assessed by the comet assay on Chinese Hamster ovary cells.

Results

Hydromethanolic extracts from Nauclea latifolia, Nauclea diderrichii and Nauclea pobeguinii exhibited a significant clastogenic/aneugenic activity without S9 mix. The hydromethanolic extract from Nauclea diderrichii was the most clastogenic/aneugenic fraction with a Minimal Active Concentration (MAC) of 23.1 μg mL⁻¹. It was submitted to a separation step leading to six main saponins identified as quinovic acid glycosides (saponins A, D, E, G, J, K). None of the isolated saponins exerted a significant clastogenic/aneugenic activity by the micronucleus assay, however a mixture made with equal quantities of each of the six saponins exhibited a direct genotoxic/clastogenic activity as assessed by both the micronucleus assay and the comet assay on Chinese Hamster Ovary cells.

Conclusion

Saponins present in the hydromethanolic extracts of Nauclea induced synergistic in vitro DNA-damage and chromosome mutations in mammalian cells. This genotoxic activity was probably due to the capacity of Nauclea saponins to reduce cell defense against oxidative stress through the inhibition of glutathione-S-transferase activity.     

生化汤提取物对正常未孕、雌激素预处理及产后小鼠离体子宫平滑肌收缩功能的影响

目的:研究生化汤不同含量水提取物对正常未孕小鼠、雌激素预处理小鼠及产后小鼠子宫平滑肌收缩功能的影响。方法:采用正常未孕小鼠、雌激素预处理小鼠及产后小鼠离体子宫标本,以Medlab生物信号采集处理系统记录生化汤水提取物(3～12 mg.mL^-1)对子宫的收缩幅度及频率的影响。结果:生化汤水提取物对正常未孕小鼠及产后小鼠子宫的影响主要表现为活动力减弱;对雌激素预处理小鼠子宫收缩频率及子宫活动力均表现为明显增强,且未见明显量效关系。结论:雌激素预处理小鼠子宫对生化汤的反应不能代表正常未孕及产后小鼠子宫的药理作用。     

Chemical composition,antimicrobial activity against Staphylococcus aureus and a pro-apoptotic effect in SGC-7901 of the essential oil from Toona sinensis (A. Juss.) Roem. leaves

Ethnopharmacological relevance

Leaves of Toona sinensis (A. Juss.) Roem. (TSL), a popular vegetable in China, have anti-inflammatory, antidoting, and worm-killing effects and are used in folk medicine for the treatment of enteritis, dysentery, carbuncles, boils, and especially abdominal tumors. Our aim was to investigate the in vitro antimicrobial activity against Staphylococcus aureus and anticancer property of the essential oil from TSL (TSL-EO), especially the pro-apoptotic effect in SGC-7901.

Materials and methods

TSL-EO obtained by hydrodistillation was analyzed by GC/MS and was tested in vitro against twenty clinically isolated strains of Staphylococcus aureus (SA 1–20), which were either methicillin-sensitive Staphylococcus aureus (MSSA) or methicillin-resistant Staphylococcus aureus (MRSA) and two standard strains viz. ATCC 25923 and ATCC 43300. The anticancer activity of TSL-EO was evaluated in vitro against HepG2, SGC7901, and HT29 through MTT assay. Moreover, the apoptosis-inducing activity of TSL-EO in SGC7901 cells was determined by Hoechst 33324 staining and flow cytometry methods. Also, the apoptosis-related proteins viz. Bax, Bcl-2 and caspase-3 were detected by western-blotting.

Results

GC–MS analysis showed that TSL-EO contained a high amount of sesquiterpenes (84.64%), including copaene (8.27%), β-caryophyllene (10.16%), caryophyllene (13.18%) and β-eudesmene (5.06%). TSL-EO inhibited the growth of both MSSA and MRSA, with the lowest MIC values of 0.125 and 1 mg/ml, respectively. Treatment with TSL-EO for 24 h could significantly suppress the viability of three different cancer cell lines (P<0.05). Furthermore, the apoptosis-inducing activity of TSL-EO in SGC7901 cells increased in a dose-dependent manner, potentially resulting from the up-regulated expression of Bax, caspase-3 and down-regulated expression of Bcl-2.

Conclusions

TSL-EO possessed antibacterial activity against Staphylococcus aureus and significant cytotoxicity against cancer cells and particularly prominent pro-apoptotic activity in SGC7901 cells. These bioactivities were probably due to the high content of sesquiterpenes. Our results suggested that TSL-EO possessed potential health benefits and could serve as a promising natural food addictive.     

In vivo analgesic activity,toxicity and phytochemical screening of the hydroalcoholic extract from the leaves of Psidium cattleianum Sabine

Ethnopharmacological relevance

Psidium cattleianum Sabine is extensively used in Brazilian traditional medicine to treat several diseases including painful disorders. Aim of the study to investigate the toxicity and the possible analgesic activities of the hydroalcoholic extract from the leaves of Psidium cattleianum Sabine (ELPCS), to support its use in folk medicine. To screen the major phytochemical constituents of this extract and evaluate their antioxidant activity.

Materials and methods

ELPCS was assessed for its antioxidant activity using the DPPH model. Its analgesic activity was examined using mouse models of acetic acid-induced writhing and hot plate paw licking models. The major phytochemical constituents of the extract were screened; their toxicity on LLC-MK2 mammalian cells was evaluated.

Results

ELPCS exhibited significant peripheral analgesic activity at doses of 60, 80, 100, 200 and 400 mg/kg in mice, but it did not display central analgesic activity and not was toxic to LLC-MK2 cell (LD₅₀>400 µg/mL). The extract exhibited free radical scavenging activity as evidenced by IC₅₀ values (15.9 µg/mL) obtained by the DPPH method. Phytochemical screening detected flavonoids, saponins, cardiac glycosides, anthraquinones, and tannins.

Conclusions

The results of the experimental studies proved the analgesic activity of ELPCS and supported the traditional use of this plant.