Bioassay-Guided Chromatographic Isolation and Identification of Antibacterial Compounds from Artemisia annua L. That Inhibit Clostridium perfringens Growth

Abstract Clostridium perfringens is the causative agent of necrotic enteritis leading to significant losses in the poultry industry. Dichloromethane and n-hexane extracts of aerial parts of Artemisia annua (Asteraceae) exhibited activity against C. perfringens with minimum inhibitory concentrations (MIC) of 185 and 270 μg/mL, respectively. Bioassay-guided fractionation of the extracts gave several active fractions (MIC between 75 and 600 μg/mL). Investigations of the most active fractions resulted in the iso ponticaepoxide (MIC between 100 and 200 μg/mL) and (+)-threo-(5E)-trideca-1,5-dien-7,9,11-triyne-3,4-diol (MIC between 400 and 800 μg/mL), the flavonols chrysosplenol D (MIC between 200 and 400 μg/mL) and casticin (slight inhibition at 800 μg/mL), and 2,4-dihydroxy-6-methoxyacetophenone (slight inhibition at 800 μg/mL). Also, the coumarin scopoletin and the sesquiterpene lactone artemisinin were isolated from active fractions but showed no inhibition of C. perfringens growth at 800 and 2000 μg/mL, respectively. Fractions containing essential oil components with camphor constituting >60% did not show inhibition of C. perfringens up to 1600 μg/mL. Extracts and some active fractions showed higher antibacterial effect than individual bioactive compounds, suggesting that synergistic effects may underlie the observed antibacterial effect. The present study adds new valuable information on the antibacterial effect of A. annua against C. perfringens.

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Antibacterial Alkaloids from Chelidonium Majus Linn (Papaveraceae) Against Clinical Isolates of Methicillin-Resistant Staphylococcus Aureus

Journal of Pharmacy & Pharmaceutical Sciences ◽

10.18433/j3d30q ◽

2009 ◽

Vol 11 (4) ◽

pp. 90 ◽

Cited By ~ 46

Author(s):

Guo Ying Zuo ◽

Fan Yan Meng ◽

Xiao Yan Hao ◽

Yun Ling Zhang ◽

Gen Chun Wang ◽

...

Keyword(s):

Staphylococcus Aureus ◽

Antibacterial Agents ◽

Methicillin Resistant Staphylococcus Aureus ◽

Antibacterial Effect ◽

Dilution Method ◽

Chelidonium Majus ◽

Methicillin Resistant ◽

Aerial Parts ◽

Bioassay Guided Fractionation ◽

Mrsa Strains

Purpose. This study describes the antibacterial effect of extracts and compounds isolated from the aerial part of Chelidonium majus Linn. (Papaveraceae) acting against clinical strains of methicillin-resistant Staphylococcus aureus (MRSA). Methods. The activities were evaluated by using the macrobroth dilution method and reported as the MICs/MBCs. Results. Bioassay-guided fractionation of the most active extract from the aerial parts (EtOAc) led to the isolation of benzo[c]phenanthridine-type alkaloids 8-hydroxydihydrosanguinarine (hhS), 8-hydroxydihydrochelerythrine (hhC), which were potently active against MRSA strains. Conclusions. The selective antibacterial activity reported in this paper for 8-hydroxylated benzo[c]phenanthridine-type alkaloids isolated from C.majus opens the possibility that they could be helpful for the developing of new antibacterial agents for treating the infection of MRSA which has created nosocomial problem worldwide.

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Evaluation of the Antidiabetic and Antibacterial Activity of Cissus sicyoides

Brazilian Archives of Biology and Technology ◽

10.1590/s1516-89132002000100004 ◽

2002 ◽

Vol 45 (1) ◽

pp. 21-25 ◽

Cited By ~ 9

Author(s):

Flavio Luis Beltrame ◽

Greisiele Lorena Pessini ◽

Dani Luce Doro ◽

Benedito Prado Dias Filho ◽

Roberto Barbosa Bazotte ◽

...

Keyword(s):

Antibacterial Activity ◽

Bacillus Subtilis ◽

Antibacterial Effect ◽

Methanolic Extract ◽

Serum Levels ◽

Diabetic Rats ◽

Antidiabetic Activity ◽

Minimal Inhibitory Concentrations ◽

Aerial Parts ◽

Bioassay Guided Fractionation

In this work we investigated the antidiabetic and antibacterial effect of Cissus sicyoides (CS) from Brazil. Diabetic rats that received water (A group) or extracts from the aerial parts of the plant (Cs group) during four weeks were employed. After this period, serum levels of glucose, cholesterol and triglycerides were measured. Glycemia was not affected by treatment with CS. However, there was an increased cholesterol and triglyceride level in Cs group. In addition, bioassay-guided fractionation of methanolic extract from aerial parts of CS was performed for isolation of antibacterial compounds.beta-Sitosterol and sitosterol-beta-D-glucopyranoside isolated showed antibacterial activity against Bacillus subtilis with minimal inhibitory concentrations (MICs) of 50 mug/ml and 100 mug/ml, respectively. In spite of popular belief, CS did not show antidiabetic activity. However, two compounds isolated from aerial parts of the plant (beta-sitosterol and sitosterol-beta-D-glucopyranoside) showed antibacterial activity.

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Identifying the Virulent Factors of Clostridium perfringens Locally Isolated from Different Species

World s Veterinary Journal ◽

10.54203/scil.2020.wvj74 ◽

2020 ◽

Vol 10 (4) ◽

pp. 617-624

Author(s):

H. A. El-Helw ◽

M. M. Taha ◽

Elham F. El-Sergany ◽

Ebtesam, E.Z. Kotb ◽

A. S Hussein ◽

...

Keyword(s):

Virulence Factors ◽

Clostridium Perfringens ◽

Animal Species ◽

Necrotic Enteritis ◽

Isolation And Identification ◽

Clostridium Perfringens Enterotoxin ◽

Virulent Strains

Clostridium perfringens incriminated in many diseases among different species of animals due to its ability to produce many virulence factors. In the current study, 135 intestinal samples were collected from different animal species of different localities in Egypt. Samples were subjected to isolation and identification (morphologically and biochemically) for obtaining Clostridium perfringens isolates (n=26, 19.25%). The PCR was carried out to elucidate the virulence factors. It was indicated that all the 26 Clostridium perfringens isolates had CPA gene and Clostridium perfringens enterotoxin (CPE gene), whereas 23% of isolates of chicken and cattle intestinal samples contained CPA, Net B, and CPE genes as virulence factors. Consequently, those isolates are highly recommended to be used in the preparation of enterotoxemia and necrotic enteritis vaccines as they are more virulent strains.

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Synergistic antibacterial combination of Lavandula latifolia Medik. essential oil with camphor

Zeitschrift für Naturforschung C ◽

10.1515/znc-2020-0051 ◽

2020 ◽

Vol 0 (0) ◽

Author(s):

Nursenem Karaca ◽

Görkem Şener ◽

Betül Demirci ◽

Fatih Demirci

Keyword(s):

Staphylococcus Aureus ◽

Essential Oil ◽

Antibacterial Effect ◽

Synergistic Effects ◽

Pharmaceutical Formulations ◽

Human Pathogens ◽

Broth Microdilution ◽

Antibacterial Effects ◽

Lavandula Latifolia

AbstractCombination of various compounds and essential oils for pharmaceutical formulations withdraw attention. In this present study, it was aimed to evaluate the in vitro potential synergistic antibacterial effect of Lavandula latifolia (spike lavender) essential oil with camphor by using the checkerboard method against the human pathogens; Staphylococcus aureus and Listeria monocytogenes. Pharmacopoeia quality L. latifolia essential oil and racemic camphor were analyzed and verified by GC-FID and GC/MS, simultaneously. In vitro antibacterial activity of essential oil and camphor (MIC range: 0.16–20 mg/mL) and standard antimicrobial clarithromycin (MIC range: 0.125–16 μg/mL) were carried out by broth microdilution against S. aureus and L. monocytogenes standard strains, respectively. Resulting antibacterial effects were evaluated for their fractional inhibitory concentrations (FICs) as antagonistic, additive and synergistic effects. The analytical results showed that the major component of essential oil was linalool (45.2%) and 1,8-cineole (25.6%). Antibacterial effects of essential oil were determined as MIC 1.25–5 mg/mL. As a result of the experiments, L. latifolia essential oil–camphor combinations were identified as “synergistic (FIC ≤ 0.5), and additive (0.5 < FIC ≤ 1)” in the respective combinations, suggesting further evaluation for formulations for potential antimicrobial applications in food and pharmaceuticals.

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Supplementing Garlic Nanohydrogel Optimized Growth, Gastrointestinal Integrity and Economics and Ameliorated Necrotic Enteritis in Broiler Chickens Using a Clostridium perfringens Challenge Model

Animals ◽

10.3390/ani11072027 ◽

2021 ◽

Vol 11 (7) ◽

pp. 2027

Author(s):

Doaa Ibrahim ◽

Tamer Ahmed Ismail ◽

Eman Khalifa ◽

Shaimaa A. Abd El-Kader ◽

Dalia Ibrahim Mohamed ◽

...

Keyword(s):

Growth Performance ◽

Clostridium Perfringens ◽

Broiler Chickens ◽

Body Weight Gain ◽

Economic Benefits ◽

Production Costs ◽

Broiler Chicks ◽

Necrotic Enteritis ◽

Feed Conversion ◽

Intestinal Lesion

Necrotic enteritis (NE) caused by Clostridium perfringens (C. perfringens) results in impaired bird growth performance and increased production costs. Nanotechnology application in the poultry industry to control NE outbreaks is still not completely clarified. Therefore, the efficacy of dietary garlic nano-hydrogel (G-NHG) on broilers growth performance, intestinal integrity, economic returns and its potency to alleviate C. perfringens levels using NE challenge model were addressed. A total of 1200 male broiler chicks (Ross 308) were assigned into six groups; four supplemented with 100, 200, 300 or 400 mg of G-NHG/kg diet and co-challenged with C. perfringens at 21, 22 and 23 d of age and two control groups fed basal diet with or without C. perfringens challenge. Over the total growing period, the 400 mg/kg G-NHG group had the most improved body weight gain and feed conversion efficiency regardless of challenge. Parallel with these results, the mRNA expression of genes encoding digestive enzymes (alpha 2A amylase (AMY2A), pancreatic lipase (PNLIP) and cholecystokinin (CCK)) and intestinal barriers (junctional adhesion molecule-2 (JAM-2), occludin and mucin-2 (Muc-2)) were increased in groups fed G-NHG at higher levels to be nearly similar to those in the unchallenged group. At 14 d post challenge, real-time PCR results revealed that inclusion of G-NHG led to a dose-dependently decrease in the C. perfringens population, thereby decreasing the birds’ intestinal lesion score and mortality rates. Using 400 mg/kg of G-NHG remarkably ameliorated the adverse effects of NE caused by C. perfringens challenge, which contributed to better growth performance of challenged birds with rational economic benefits.

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Evaluation of different antigenic preparations against necrotic enteritis in broiler birds using a novel Clostridium perfringens type G strain

Anaerobe ◽

10.1016/j.anaerobe.2021.102377 ◽

2021 ◽

pp. 102377

Author(s):

Hiramoni Sarmah ◽

Ritam Hazarika ◽

Shantonu Tamuly ◽

Pankaj Deka ◽

Seeralan Manoharan ◽

...

Keyword(s):

Clostridium Perfringens ◽

Necrotic Enteritis

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Isolation and Identification of Two Potent Phytotoxic Substances from Afzelia xylocarpa for Controlling Weeds

Applied Sciences ◽

10.3390/app11083542 ◽

2021 ◽

Vol 11 (8) ◽

pp. 3542

Author(s):

Ramida Krumsri ◽

Kaori Ozaki ◽

Toshiaki Teruya ◽

Hisashi Kato-Noguchi

Keyword(s):

Ferulic Acid ◽

Vanillic Acid ◽

Synergistic Effects ◽

Phleum Pratense ◽

Lepidium Sativum ◽

Dependent Manner ◽

Leaf Extracts ◽

Isolation And Identification ◽

Phytotoxic Activity ◽

Seedling Length

Phytotoxic substances released from plants are considered eco-friendly alternatives for controlling weeds in agricultural production. In this study, the leaves of Afzelia xylocarpa (Kurz) Craib. were investigated for biological activity, and their active substances were determined. Extracts of A. xylocarpa leaf exhibited concentration-dependent phytotoxic activity against the seedling length of Lepidium sativum L., Medicago sativa L., Phleum pratense L., and Echinochloa crus-galli (L.) P. Beauv. Bioassay-guided fractionation of the A. xylocarpa leaf extracts led to isolating and identifying two compounds: vanillic acid and trans-ferulic acid. Both compounds were applied to four model plants using different concentrations. The results showed both compounds significantly inhibited the model plants’ seedling length in a species-dependent manner (p < 0.05). The phytotoxic effects of trans-ferulic acid (IC50 = 0.42 to 2.43 mM) on the model plants were much greater than that of vanillic acid (IC50 = 0.73 to 3.17 mM) and P. pratense was the most sensitive to both compounds. In addition, the application of an equimolar (0.3 mM) mixture of vanillic acid and trans-ferulic acid showed the synergistic effects of the phytotoxic activity against the root length of P. pratense and L. sativum. These results suggest that the leaves of A. xylocarpa and its phytotoxic compounds could be used as a natural source of herbicides.

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Recurring Necrotic Enteritis Outbreaks in Commercial Broiler Chicken Flocks Strongly Influence Toxin Gene Carriage and Species Richness in the Resident Clostridium perfringens Population

Frontiers in Microbiology ◽

10.3389/fmicb.2017.00881 ◽

2017 ◽

Vol 8 ◽

Cited By ~ 5

Author(s):

Marie-Lou Gaucher ◽

Gabriel G. Perron ◽

Julie Arsenault ◽

Ann Letellier ◽

Martine Boulianne ◽

...

Keyword(s):

Species Richness ◽

Clostridium Perfringens ◽

Broiler Chicken ◽

Toxin Gene ◽

Necrotic Enteritis ◽

Commercial Broiler

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Quantitative Detection of Clostridium perfringens in the Broiler Fowl Gastrointestinal Tract by Real-Time PCR

Applied and Environmental Microbiology ◽

10.1128/aem.71.7.3911-3916.2005 ◽

2005 ◽

Vol 71 (7) ◽

pp. 3911-3916 ◽

Cited By ~ 71

Author(s):

Mark G. Wise ◽

Gregory R. Siragusa

Keyword(s):

Gastrointestinal Tract ◽

Real Time ◽

Clostridium Perfringens ◽

Real Time Pcr ◽

Limit Of Detection ◽

Quantitative Detection ◽

Necrotic Enteritis ◽

Analytical Sensitivity ◽

Quantitative Real Time Pcr ◽

Pcr Inhibitor

ABSTRACT Strains of Clostridium perfringens are a frequent cause of food-borne disease and gas gangrene and are also associated with necrotic enteritis in chickens. To detect and quantify the levels of C. perfringens in the chicken gastrointestinal tract, a quantitative real-time PCR assay utilizing a fluorogenic, hydrolysis-type probe was developed and utilized to assay material retrieved from the broiler chicken cecum and ileum. Primers and probe were selected following an alignment of 16S rDNA sequences from members of cluster I of the genus Clostridium, and proved to be specific for C. perfringens. The assay could detect approximately 50 fg of C. perfringens genomic DNA and approximately 20 cells in pure culture. Measurements of the analytical sensitivity determined with spiked intestinal contents indicated that the consistent limit of detection with ileal samples was approximately 102 CFU/g of ileal material, but only about 104 CFU/g of cecal samples. The decreased sensitivity with the cecal samples was due to the presence of an unidentified chemical PCR inhibitor(s) in the cecal DNA purifications. The assay was utilized to rapidly detect and quantify C. perfringens levels in the gut tract of broiler chickens reared without supplementary growth-promoting antibiotics that manifested symptoms of necrotic enteritis. The results illustrated that quantitative real-time PCR correlates well with quantification via standard plate counts in samples taken from the ileal region of the gastrointestinal tract.

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