Category: Moms

Anti-microbial activity

Anti-microbial activity

Anti-mifrobial were incubated with S. Anti-microbial activity Institució Maca root and fertility de Xctivity i Estudis Avancats ICREA, Barcelona, Anti-microbizl Clivia M. This fact has also been verified in other clinics around all over world. This is an open-access article distributed under the terms of the Creative Commons Attribution License CC BY. d Explain or demonstrate how to use Whatman antibiotic assay paper discs to assess microbial action. Anti-microbial activity

Anti-microbial activity -

Broad spectrum—antibiotics act against gram positive and gram negative bacteria, for example amoxicillin. It is based on the chemical and physical properties of their cell walls. Primarily, it detects peptidoglycan, which is present in a thick layer in Gram positive bacteria.

The Gram stain is almost always the first step in the identification of a bacterial organism, and is the default stain performed by laboratories over a sample when no specific culture is referred. While Gram staining is a valuable diagnostic tool in both clinical and research settings, not all bacteria can be definitively classified by this technique, thus forming Gram-variable and Gram-indeterminate groups as well.

A broad spectrum antibiotic acts against both Gram-positive and Gram-negative bacteria, in contrast to a narrow spectrum antibiotic, which is effective against specific families of bacteria. An example of a commonly used broad-spectrum antibiotic is ampicillin. Broad spectrum antibiotics are properly used in the following medical situations: empirically i.

This occurs, for example, in meningitis, where the patient can become fatally ill within hours if broad-spectrum antibiotics are not initiated.

Broad spectrum antibiotics are also used for drug resistant bacteria that do not respond to other, more narrow spectrum antibiotics and in the case of superinfections, where there are multiple types of bacteria causing illness, thus warranting either a broad-spectrum antibiotic or combination antibiotic therapy.

Following a year hiatus in discovering new classes of antibacterial compounds, three new classes of antibacterial antibiotics have been brought into clinical use: cyclic lipopeptides such as daptomycin , glycylcyclines such as tigecycline , and oxazolidinones such as linezolid.

Search site Search Search. These bioactive compounds are used as a starting point for antibiotics synthesis in order to treat infectious diseases [ 22 ]. The crude extract of Polygonum persicaria , Rumex hastatus , Rumex dentatus , Rumex nepalensis , Polygonum plebejum , and Rheum australe have antibacterial and antifungal activities, inhibit the growth of C.

frundii , E. coli , E. aerogenes , and S. aureus [ 23 ]. The n-hexane extracts of Calotropis gigantean have no antibacterial and antifungal activities, against pathogenic microbes.

But, its ethyl-acetate fraction has inhibitory effects on some bacteria and fungi except T. rubrum [ 24 ]. Calotropis gigantean crude extracts show promising antifungal activity against Candida albicans , Aspergillus niger , Aspergillus ochraceus , Aspergillus ustus , and Rizopus oryzae pathogenic fungi in Asia [ 21 ].

In another study, an ethanolic extract of Plumbago zeylanica root exhibit good antimicrobial activities against V. cholerae , E. coli , P. aeruginosa , Curvularia lunata , Colletotrichum corchori , and Fusarium equiseti [ 25 ]. The aqueous leaf extract of Euphorbia hirta , Erythrophleum suaveolens , and the methanolic leaf extract of Thevetia peruviana has an antibacterial effect against Extended Spectrum Beta-Lactamase ESBL producing bacteria such as E.

coli , Pseudomonas , K. pneumonia , Methicillin-Resistant Staphylococcus aureus MRSA , Salmonella and Proteus [ 26 — 28 ].

The few studies conducted on the aqueous and hydro-alcoholic extracts from different plants implicate antibacterial effect on multidrug-resistant bacteria including MRSA and ESBL producing bacteria [ 29 , 30 ].

Moreover, some plants may exhibit broad-spectrum of antimicrobial effects, which possibly control the impairments associated with multidrug-resistant microbes [ 31 ].

In this regard, traditional medicinal plants TMP , are the most valuable source of new bioactive chemical entities due to their ecological biodiversity and diverse chemical endowment within each species [ 31 ].

Thus, P. aethiopica , E. depauperata , C. englerianum , L. adoensis , C. pustulatus , D. abyssinicus are TMP used by communities for the treatment of different disease caused by notorious infectious agents.

Yet, there are limited detailed and thorough examinations of these plants for their potential role as antimicrobials and phytochemical entities as therapeutic agents for MDR bacteria and pathogenic fungi.

In vitro , an experimental study was carried out to evaluate the antimicrobial status and phytochemical screening of P. abyssinicus against clinical isolate multidrug-resistant microbes and their respective reference strains.

In this study, the selected medicinal plants were collected from Berbere district in Bale Zone, being guided by the local traditional healers and resident farmers who utilize these traditional medicines.

It is located about km away from the Ethiopia capital city Addis Ababa in the southeast direction, in Bale Zone of Oromia Regional State.

The district has 17 peasant associations, where the northern and southern parts are more of highlands and lowlands respectively. Out of the 17 peasant associations, the medicinal plants were collected from seven peasant associations of Berbere district based on the profundity of traditional knowledge generated during the pilot study interview; namely Burkitu Lippia adoensis , Chekata Polysphaeria aethiopica , Darasa Cucumis pustulatus , Gabe Keka Discopodium penninervium , Gora Heddo Rumex abyssinicus , Harawa Cirsium englerianum , and Sirrima Euphorbia depauperate.

Berbere district is far away from the Zonal city Bale-Robe and gets less coverage of infrastructures in terms of hospital, transport, electrical power supplies, and other social security rendering institutions.

Until the conclusion of these fieldworks plant collection and interviews , there is only one health center in the district and the residents are imposed to use medicinal plants for the treatment of skin diseases, diabetes, Urinary Tract Infections, hepatitis, Sexually Transmitted Diseases, cancer, hypertension, sexual impotence, and as contraceptives.

Of which, P. abyssinicus are used by healers for the treatment of bacterial infection such as eczema, gonorrhea, syphilis, pneumonia, scabies, skin infection, and superficial mycosis. The leaf, bark, and root parts of seven plants, P. abyssinicus were collected and screened for their phytochemical bioactive ingredients and evaluated for antimicrobial effects on common human pathogens bacteria and fungi.

The collected plant materials were handled with standard storage protocols and transported being wrapped by plastic sheets. Authentication of each plant sample was carried out and deposited at the National Herbarium, Department of Biology, College of Natural and Computational Sciences, Addis Ababa University AAU , Addis Ababa, Ethiopia.

The collected plant parts were washed thoroughly under running tap water and rinsed in distilled water, air-dried at room temperature under shade 23—27°C , and reduced to appropriate size. The powdered plant materials were packed in a plastic bag and kept until extraction.

The powdered plant materials were weighed by sensitive digital weighing balance and a total of 1. The extraction process was facilitated by occasional shaking. After three days of frequent agitation, the extract was separated from the marc using gauze and the resulting liquid was filtered using Whatman filter paper No.

The residue was re-macerated and repeated three times to exhaustively extract the plant material. The filtrates obtained from the successive maceration were concentrated under reduced pressure using a rotary evaporator Hamato, Japan followed by a hot air oven Medit-Medizin Technik, Germany set at 40°C.

Extraction was repeated seven times and the filtrates of all portions were combined in one vessel. The organic solvent was removed by evaporation using a Rotary evaporator at 40°C.

After the removal of the organic solvent, the aqueous residue was placed in a lyophilizer until non-polar solvents were removed and the extracts became dried.

The extract was further concentrated to dryness by freeze-drying using a lyophilizer Labfreez, China. The weight of the dry extract was expressed as a percentage of the total mass of dry plant matter to determine the percentage yield [ 18 , 31 ].

Each extract of methanol and aqueous was tested for the growth of microbes. This was carried out by inoculating 0. The fungal growth was tested on Sabouraud Dextrose Agar for 4 days being incubated at 25°C. The plates were observed for growth.

The absence of growth in the extracts after incubation indicates sterility and evaluated for antimicrobial activity as indicated in CLSI guidelines [ 32 ]. Nutrient Agar, TSY Broth, MacConkey, Muller Hinton Agar MHA , Muller Hinton Broth MHB , Blood Agar BA , Mannitol Salt Agar MSA , Chocolate Agar, and biochemical reagents for bacteria and Sabouraud Dextrose Agar SDA for fungi were obtained from the Department of Medical Microbiology, Immunology and Parasitology, Addis Ababa University and Tikur Anbessa Specialized Hospital TASH Bacteriology and Pharmacology Units.

The reference bacterial species; American type cell culture ATCC of Escherichia coli ATCC , Klebsiella pneumoniae ATCC , Streptococcus pyogenes ATCC , and Staphylococcus aureus ATCC and their respective MDR strains, and Trichophyton mentagrophytes ATCC and Candida albicans ATCC were collected from Bacteriology Unit of the Microbiology Laboratory of TASH.

All laboratory works were performed according to CLSI guidelines [ 33 ]. The antimicrobial discs used for this research were the products of Liofilchem® Inc. Liofilchem, Clinical and Industrial Microbiology Company produced in mentagrophytes as per the CLSI guideline protocols [ 32 , 33 ].

Multidrug-resistant gram-negative and gram-positive bacteria were isolated from three different samples urine, throat swab, and blood. Few colonies 3 to 5 of similar morphology of the respective bacteria were transferred with a sterile inoculating loop to a liquid medium until adequate growth of turbidity with McFarland in 0.

Then the bacterial suspension was streaked onto MHA plates using a sterile swab in such a way as to ensure thorough coverage of the plates and a uniform thick lawn of growth following incubation. The susceptibilities of clinical isolates were tested by using the MHA impregnated with a range of antimicrobial agents.

Dilutions of overnight broth cultures were inoculated onto antibiotic-containing plates to yield final inoculums of approximately 10 6 CFU per spot according to CLSI protocol guidelines for Enterobacteriaceae.

Selected multidrug-resistant, K. pneumoniae , and E. coli were screened for their resistance for more than two different classes of antibiotics following the disk diffusion method protocol as in the CLSI guidelines and WHO recommendations [ 33 ].

After preliminary screening of plants for their antimicrobial activity, those which revealed potent antimicrobial effects were further tested to determine MIC and MBC against multidrug-resistant gram-negative, gram-positive, and Candida species. It was determined by the MHB broth micro-dilution method.

Each of the well microtiter plates was liquated with 50μl of MHB; while μl of MHB was added to the 10th well sterile control. A serial two-fold dilution was performed by transferring 50μl of the suspension to the subsequent wells until the 8th well as per the protocol and procedure is given by CLSI guidelines and the modified protocol of Wiegand [ 32 ].

About 0. Bacterial cell viability and MIC values were determined by turbidity test. The lowest concentration of the extract with clear suspension was considered as the MIC values.

The lowest concentration of the extract in the post-incubation suspensions which did not harbor any bacterial growth upon spotting on MHA after overnight incubation at 37°C was considered as the MBC values.

The method adopted for the fungus was the same as that of the bacteria. Instead of nutrient agar, Sabouraud dextrose agar was used.

The inoculated medium was incubated at 25°C for C. The test was performed in triplicates alongside antibiotics ciprofloxacin 5μg as a positive control at a concentration of 0.

Ketoconazole was used as a positive control at a concentration of 0. Standard preliminary phytochemical qualitative analysis of the extract was carried out for the various plant constituents and screened for the presence or absence of biologically active compounds or secondary metabolites using standard procedures.

Around mg plant material was boiled in 10mL methanol and filtered. The brownish-red precipitate was taken as a piece of evidence for the presence of alkaloids. Towards the end, an alkaloids test was applied on few plant materials by diluting 2. Then, 1ml of the filtrate was added to 1ml of dilute ammonia.

Finally, 1ml of chloroform CHCl 3 was added and shaken gently to reveal the alkaloid base. Then, the filtrates were allowed to reacted with 2. The presence of pink, red, or violet color in ammonia solution in the lower phase indicates a positive result.

Test for cardiac glycosides Keller-Kiliani test : About 1. The reddish-brown color at the interface indicates the presence of a steroidal ring, the glycone portion of cardiac glycosides.

Test for flavonoids : About 7. A yellowish coloration indicates the presence of flavonoids. The crude extract of the plant material was treated with 3 to 4 drops of ferric chloride solution, or dissolving 5mg of dry extract in 0.

The formation of bluish-black color indicates the presence of phenolic compounds. Test for saponin Frothing test : About 2.

Samples showing froth were warmed. Persistent foam formation indicates the presence of saponin. Two to three drops of 0. Test for terpenoids Salkowski test : About 0. The gray color indicates the presence of terpenoids. The triplicate data reading values of inhibition zones in diameter and concentration values MIC and MBC were analyzed using Statistical Packages for Social Sciences SPSS software, Window version 22 according to CLSI.

Each experimental value was expressed in terms of mean SD mean and standard deviation. Significance in the difference between the two groups was tested by Student t-test, assessed by comparing the corresponding p-value of the test. The Research Ethics Committee under the Office of Vice President for Research of Madda Walabu University, the only University in Bale Zone with its main campus in Bale-Robe and the Berbere district administrative office bilaterally approved the fieldwork of this study.

On the other hand, as the research was joint collaborative research between Madda Walabu and Kotebe Metropolitan Universities, the Research Ethics Committee of Menelik Medical and Health Science College, Kotebe Metropolitan University in Addis Ababa, Ethiopia granted an ethical approval letter for the study to make use of the patient samples collected at its disposal and other two hospitals in Addis Ababa under its supervision Ref.

The ethically cleared study, from which we extracted samples from the human subjects urine, throat swab, and blood samples was entitled "Molecular characterization of multidrug-resistant microbes among antenatal care attending pregnant women in Addis Ababa" and principally carried out by one of the co-authors of this study.

His research team underwent all the standard procedures of ethical clearance, where informed written consent was obtained from all study participants prior to study participation.

For those participants who were below the legal consenting age Furthermore, the current study followed all the standard procedures of random representative sample selection simple random sampling for the inclusion of unbiased patient samples collected from Addis Ababa into the study subjects, after we had had full permissions from Menelik Tertiary Level Hospital, Kotebe Metropolitan University.

The dried and powdered leaves of P. aethiopica , C. englerianum , E. depauperata , L. penninervium , and root of C. pustulatus and R. They had shown promising antimicrobial effects on E. coli , K. pneumoniae , S.

aureus , S. pyogenes , C. albicans , and T. Almost all extracts showed antibacterial activity against two or more multidrug-resistant and reference strains of human pathogenic bacteria. The extract of E.

depauperata , R. abyssinicus , and L. adoensis showed the broadest spectrum of action as they inhibited the growth of E. aureus , and S. pyogenes with a zone of inhibition in diameter ranging from 29mm Table 1. The methanolic extract of C.

adoensis , and E. depauperate showed inhibition zone in diameter values of 28mm, 27mm, and 26mm on the MRSA, respectively.

The novel antimicrobial activity was observed from the methanolic extract of C. englerianum against the S. It had an inhibition zone of 29mm against both strains collected from the sample and reference strain. In addition, S.

aureus had a remarkable inhibition zone Fig 1. The methanolic extract obtained from E. depauperata revealed appreciable antibacterial activity against MSSA and MRSA with an inhibition zone of 27mm and 26mm, in that order Table 1.

On the other hand, MDR gram-negative pathogenic bacteria, K. pneumoniae strains were the second susceptible bacteria. It had an inhibition zone in diameter ranging from 19mm to 29mm.

Similarly, another MDR gram-negative pathogenic bacteria, E. coli had shown 15mm, 22mm, and 27mm inhibition zone in diameter when the extract obtained from E.

englerianum , and D. penninervium were applied, respectively Table 1. However, the extract from P. aethiopica failed to show an observable zone of inhibition against the MDR E.

coli bacteria tested Fig 2. The MIC value of plant extracts against the tested Gram-positive bacteria ranged from 1. The lowest MIC and MBC values were observed in the case of S. pyogenes , which were the most inactivated bacteria species by most methanolic extracts.

Also, these findings divulge remarkable growth inhibition of both MRSA, with MIC and MBC value ranges of 4. The second most inactivated bacteria were clinical isolate MDR and reference strain of K.

pneumoniae , with MIC and MBC value ranges of 2. The minimum bactericidal concentration test was directly proportional to its respective MIC value. The bacteria species inhibited at the lower concentration of the plant extracts also showed no growth in subculture media with relatively lower concentration.

This study revealed that the methanolic extracts obtained from C. abyssinicus demonstrated remarkable antifungal effects against C. mentagrophytes Table 1. The methanolic extract of R. abyssinicus showed a promising antifungal effect over others. Likewise, extracts of D.

penninervium and C. No antifungal activity was observed with extracts of P. aethiopica against C. mentagrophytes Table 2. The highest yield was observed in C. In terms of the qualitative phytochemical investigation of the medicinal plants, the medicinal plants extract had different phytochemicals constituents such as saponins, tannins, alkaloids, terpenoids, anthraquinones, phenolic compounds, cardiac glycosides, and flavonoids Table 4.

These bioactive compounds are naturally occurring in most extracts and identified to possess bactericidal or fungicidal properties on the tested human pathogens. Biomedical science has exploited plants as the potential sources of drugs to prevent and cure human diseases.

The World Health Organization has recognized antimicrobial resistance as a global health security threat that requires action across government sectors and society as a whole [ 4 ]. It is obvious that the increasing occurrences of MDR microbes ameliorate socioeconomic crises and dwindle public health status worldwide.

These MDR microbes compromise the success of many currently accessible and affordable antimicrobials on the shelf, especially in developing countries. It is for these reasons that the search for new antimicrobials from medicinal plants to combat the growing threat from those pathogenic microbes, MDR bacteria, and fungi.

This could be demonstrated by almost all tested medicinal plants in this study, where the extracts from the majority of them showed a glimpse of prospective new drug discoveries to cure and control diseases caused by MDR pathogenic microbes Figs 1 and 2.

These findings corroborate previous studies that the therapeutic agents derived from plants are used as an important surrogate, alternative, or complementary treatment of infectious diseases [ 16 — 18 ].

This current study revealed that all species of plants tested had shown antibacterial activity against S. pyogenes , as this bacterium was the most susceptible species of all tested human pathogenic bacteria. Besides, most of the extracts had a remarkable and pronounced inhibitory effect on MRSA, next to S.

Moreover, multidrug-resistant and the reference strains of K. pneumoniae were found to be the most sensitive gram-negative bacteria by the majority of the tested plant extracts. Implausible, both multidrug-resistant and reference strains of E. coli were the most unresponsive strain of all tested bacteria species.

These findings agree with several research reports [ 25 , 37 ]. This demonstrates that gram-positive bacteria S. aureus and S. pyogenes are more sensitive to TMP extracts tested than the gram-negative E. Unlike the current study result on E. coli , many studies have shown a glimmer of hope as medicinal plants are endowed with many different bioactive compounds that potentially inhibit the growth of this human bacterial pathogen [ 19 ].

On the other hand, researchers demonstrated that E. coli has developed multidrug resistance to many currently available and affordable antibiotics on the market [ 20 , 27 ]. Thus, genetic makeup to E. coli to produce multidrug resistance was favorable to impede the antibiotic effectiveness through impeding permeability and enhancing efflux pump [ 16 , 19 ].

Consequently, searching for antibacterial activity of the secondary metabolites from medicinal plants is a spotlight lately [ 24 , 29 ]. In this regard, drug discovery from medicinal plants is a near-future hope to overcome socioeconomic problems and long-term negative impacts as a result of multidrug-resistant bacteria, including MRSA, E.

coli , and K. pneumoniae [ 25 , 37 ]. Regarding pathogenic fungi of humans in the current study, the presence of bioactive compounds in traditional medicinal plants inhibited the growth of C. A study conducted elsewhere by Shumaia Parvin reported that a selected extract of C.

gigantean had inhibitory potential on all species of human pathogenic bacteria and fungi except T. Another study conducted on ethanolic extract of P. zeylanica root also reported exhibiting appreciable antimicrobial activities against V. aeruginosa , Curvularia lunata , Colletotrichum corchori , and Fusarium equiseti [ 21 , 22 ].

Overall, the current study has shown that the human fungal pathogens were relatively tolerant to many of the tested extracts as compared to the human pathogenic bacteria.

Also, among the pathogenic fungi, the current findings demonstrated that C. albicans was less susceptible to the extracts as compared to the cutaneous mycosis causing fungal agent T. This might be due to the presence of different bioactive entities that potentially inhibit the growth of cutaneous mycosis, like that of the multidrug-resistant bacteria.

Bacteria often activigy Anti-microbial activity polymicrobial communities Anti-mixrobial they compete for Anti-microbial activity resources. Intrinsic to this competition Protein benefits the ability GI values some species Anto-microbial inhibit acgivity kill their competitors. This phenomenon is pervasive throughout the human body where commensal bacteria block the colonization of incoming microorganisms. In this regard, molecular epidemiological and microbiota-based studies suggest that species-specific interactions play a critical role in the prevention of nasal colonization of the opportunistic pathogen Staphylococcus aureus. Despite this, S. The Antim-icrobial activity of plant extracts and phytochemicals Anti-microbial activity evaluated with antibiotic susceptible and resistant activkty. In addition, the Anti-nicrobial synergistic Anti-micribial when associated with antibiotics were studied. Extracts from Maca root and fertility following plants were utilized: Achillea millifolium yarrowCaryophyllus aromaticus cloveMelissa offficinalis lemon-balmOcimun basilucum basilPsidium guajava guavaPunica granatum pomegranateRosmarinus officinalis rosemarySalvia officinalis sageSyzygyum joabolanum jambolan and Thymus vulgaris thyme. The phytochemicals benzoic acid, cinnamic acid, eugenol and farnesol were also utilized. The highest antimicrobial potentials were observed for the extracts of Caryophyllus aromaticus and Syzygyum joabolanum, which inhibited

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