Biofilm-Inhibitory Activity of Wild Mushroom Extracts against Pathogenic Bacteria

Objective This study aims to investigate the bacterial biofilm-inhibitory effect of mushroom extracts. Methods Mushrooms were collected from Arabuko-Sokoke and Kakamega forests and identified using morphological and molecular approaches. Auricularia auricula-judae, Microporus xanthopus, Termitomyces umkowaani, Trametes elegans, and Trametes versicolor were extracted by chloroform, 70% ethanol, and hot water. Extracts were tested against Escherichia coli, Pseudomonas aeruginosa, and Staphylococcus aureus (ATCC25923). Data were analyzed using SPSS ver. 20.0. Results Chloroform, 70% ethanol, and hot water extracts of A. auricula-judae (50 μg/mL) showed statistically significant antibiofilm activities against P. aeruginosa, E. coli, and S. aureus (p ≤ 0.05). M. xanthopus extracts (250 μg/mL) revealed significantly significant antibiofilm activities against each test bacterium (p ≤ 0.05). All extracts of T. umkowaani (250 μg/mL) exhibited statistically significant antibiofilm activities against S. aureus only (p ≤ 0.05). Chloroform extract of T. elegans (250 μg/mL) showed the best antibiofilm activity (69.75 ± 0.01%) against S. aureus. All T. versicolor extracts (250 μg/mL) indicated the best antibiofilm activities against S. aureus. Conclusions Being the first study of its kind to be conducted in Kenya, it added a novel concept to the body of knowledge already known about medical biotechnology research. It offers a fresh understanding of the various varieties of mushrooms found in Kenya, their potential biological function in the production of drugs, particularly those that combat drug resistance, and perhaps even a peek at their bioactive elements. Wild mushrooms, a hidden gem, might help to reopen the pipeline of new antibiotics that have been on the decline. However, further research is required to determine the potential mechanism(s) of action of the extracts that are in charge of the apparent antibiofilm activity.


Introduction
Bacterial bioflms are collections of bacterial cells that are adhered to surfaces and/or to one another and enclosed in an extracellular matrix that the bacteria have created on their own [1,2].Bacteria that produce bioflms are extremely resistant to antimicrobial treatments [3,4].Te damaging efects of antibacterial agents and environmental stresses are warded of by bacterial bioflm.Due to the fact that most antibiotics are unable to pass through the bioflm's protective coating, it has a higher resistance (10-100 times) than the planktonic bacterial cells it is made of [1,5].Understanding bioflms' secret and how they develop antimicrobial resistance has recently drawn more interest from researchers [6].Te creation of new therapeutic approaches is necessary given the quick dissemination and ongoing generation of infections and bioflms that are resistant to antibiotics [7,8].Te need for alternatives is being fueled by the growing disparity between the advent of bacteria resistant to antibiotics and the paucity of newly developed therapeutic methods.Te search for efective screening and fnding promising antibioflm chemicals from natural sources is constantly expanding [9][10][11].Studies have been concentrating on natural secondary metabolites acting as nontoxic inhibitors of the quorum-sensing process within the bioflms to control infections with no encouraging fndings against resistant strains [12].
Te successful prevention and efcient treatment of bacterial infections are currently threatened globally by antibiotic resistance [13][14][15].Clinical and therapeutic results have been negatively impacted by antibiotic resistance, with efects ranging from treatment failures to greater rates of morbidity and mortality as well as high healthcare expenditures [16].Despite the fact that numerous steps have been taken in recent years to address this problem, the trend of worldwide antimicrobial resistance has not changed as of yet.Te primary causes of the emergence of antimicrobial resistance are the inappropriate and excessive use of various antibacterial agents for various objectives.Target alteration, extensive antibiotics-degrading enzyme production, porin alteration, efux pump overexpression, spontaneous evolution, bacterial mutation, and horizontal gene transfer are other important factors in the development of antimicrobial resistance [13,17].Te need for novel antibiotics and other antimicrobials is still urgent in the fght against bacterial infections by humans [14].Antibiotic-resistance catastrophe is looming due to this worrying confuence of increasing antibiotic resistance and declining innovation.In the worst-case scenario, most bacterial illnesses would once again be resistant to therapeutic treatment, sending humanity back to the preantibiotic age [15].Since natural products are a signifcant source of chemical variety and have given signifcant therapeutic agents for many bacterial infections, there has been a rise in interest in the mechanisms behind their actions in recent years.Mushrooms have long been valued for their renowned source of products with a variety of bioactivities, from antibacterial to antiviral, cytotoxic, anti-infammatory, antifungal, or antioxidant, and they may be a useful source in the search for new bioactive extracts to inhibit bioflm production [18].Giving researchers more money will speed up and expand screening campaigns, signifcantly increasing the chance that an antibiotic will be discovered [15].
Fungal species are already providing a wide range of potential for bioprospecting and downstream applications [19].To address issues with bioflm development, systematic exploration and evaluation of bioactive substances from wild mushrooms are thought to be quite benefcial [16,20].Finding and analysing bioactive components in mushrooms are highly benefcial for reducing the growth of bioflms [21].In this regard, the identifcation of antibioflm chemicals in wild mushrooms may herald a new era in the fght against microbial disease through the development of more efective methods for dealing with drug-resistant microorganisms [22].However, no research has been done to date to look for antibioflm bioactive components in these fve wild mushrooms from the Kenyan woodlands of Arabuko-Sokoke and Kakamega.Terefore, the aim of this work is to investigate the bioactive substances found in various wild mushrooms from Kenya by assessing their bioflm-inhibitory efects against three harmful bacteria.

Wild Mushrooms Collection.
Te wild mushrooms growing on tree barks and other substrates (wood, soil, and leaf litter) were randomly collected and handled carefully, wrapped in aluminum foil, and transported to the laboratory.

Morphological and Molecular Identifcation of Specimens.
Specimens were identifed using spore print color (i.e., white, black, brown, pink, and purple) and macroscopic and microscopic (i.e., shape and size of basidiospores, basidia, cystidia, and generative hyphae) methods as well as by comparing their morphological characteristics to Species Fungorum and related literature.Morphological parameters of the mushrooms such as the color and shape of the cap; the color and shape of the stipe; and the size and shape of the gill are some of the parameters used for identifcation.Te pileal margin and pileal surface, stipe location, stipe base, and gill margin were also used for the identifcation.Moreover, other morphological observations of the mushrooms, such as the structure of the cap; margin and location of the gills; location, surface, and shape of the stipe; and margin, shape, and surface of the pileus, were recorded.Furthermore, other characteristics such as the ornamentation of the surfaces of the pileus and stipe, the presence and absence of an annulus on the stipe, and the presence or absence of volva at the base of the stipe were also used to describe and identify the mushrooms (Table1).Finally, specimens were dried in an electric drying oven at 40 to 50 °C for 168 h. and were kept for further analyses [23,24].
Genomic DNA was extracted from the dried fruiting body of mushrooms using the cetyl trimethyl ammonium bromide (CTAB) method [25].Highly conserved regions of fungal rDNA-specifc genes (i.e., ITS1 and ITS4) were amplifed using the PCR amplifcation method [26].During PCR amplifcation, ITS1-F (5′-CTT GGT CAT TTA GAG GAA GTA A-3′) and ITS-4 (5′-TCC TCC GCT TAT TGA TAT GC-3′) set of forward and reverse primers were used, respectively (Table 2).Te amplifcation reaction volume (100 μL) contained 2.5 μL of 80-100 ng of specifc genes (ITS1 and ITS4), 1.5 μL of sterile Milli Q water, 1 μL of 20 pmol of each primer, and 20 μL of OneTaq Quick-Load 2x master mix (containing 0.25 mM each dNTPs, 2 mM � 0.19 g MgCl 2 , and Taq DNA polymerase).Control reactions without the DNA template were also prepared.Te PCR was performed in the following conditions: initial denaturation at 94 °C for 5 min followed by 30 cycles of denaturation at 94 °C for 1 min, annealing for 1 min at 52 °C, initial extension for 1 min at 72 °C, and fnal extension of 10 min at 72 °C, followed by cooling at 4 °C until the samples were recovered.Amplifed PCR products were separated using electrophoresis and visualized under UV light.Te presence and the amount of each DNA PCR product were estimated by comparing it against the ethidium bromide fuorescence intensity of control of 1 kb DNA ladder.Evidence-Based Complementary and Alternative Medicine

Study Design for Antibioflm Activities of Mushroom
Extracts.Te antibioflm activities of the extracts were determined by microtiter plate assay [31,32].Escherichia coli (Clinical isolate), Pseudomonas aeruginosa (Clinical isolate), and Staphylococcusaureus (ATCC25923) were inoculated to 5 mL Mueller Hinton broth (MHB) test tubes and incubated at 37 °C for 18 h.Te cultures of each bacterium were diluted 100-fold (1 : 100) with a fresh MHB to obtain OD 620 of 1.00 (1 × 10 8 CFU/mL).20 mg/mL of each mushroom extract was dissolved in dimethyl sulfoxide (DMSO).A sterile 100 μL MHB was added to the wells.From the 20 mg/mL extract, 25 μL (500 μg/mL) was dispensed to the #1 Well of rows A, B, and C of the 96 wells of the microtiter plate.Ten, the mixture of the broth and the extract was serially diluted from Well #1 up to Well #4 (the concentration of the extract ranged from 500 μg/mL Well #1 to 62.5 μg/mL Well #4).Te wells of the other three rows (rows D, E, and F) within the same microtiter plate were flled with 100 μL of MHB and 100 μL of each bacterium culture without the extracts as a negative control.
Te wells of the remaining two rows (rows G and H) were flled with 200 μL Mueller Hinton broth only as quality control.Te microtiter plates were incubated at 37 °C for 48 h.After incubation, 100 μL of planktonic bacterial cells was transferred to a new 96-well cell culture plate, and the OD 620 was measured.Te antibacterial efects of the extracts on the planktonic cells of the tested bacteria were investigated by calculating the relative amounts of planktonic cells of the tested bacteria and comparing their optical density (OD) values against their counterparts (negative controls).Te contents of each well were discarded and washed three times with sterile distilled water to remove nonadherent cells and air-dried for 15 min to visualize the bioflm formation.Te microtiter plate wells were fxed using 200 μL of 2% sodium acetate at room temperature for 10 min, and the sodium acetate solution was discarded and air-dried.
A 200 μL of 0.10% crystal violet (CV) solution was added to the wells and waited for 10 min at room temperature, and the excess stain was rinsed of with distilled water.Subsequently, the microtiter plates were vigorously tapped on a paper towel to remove any excess liquid and air-dried.Te attached and stained bacterial cells (if any) were solubilized with 200 μL of glacial acetic acid for 15 min at room temperature [33].About 200 μL of the CV and glacial acetic acid mixed solution in the wells were then transferred to other microtiter plate wells.Bioflm formation (adherence) was quantifed by measuring the optical density of the CV and glacial acetic acid mixed solution at 620 nm using a microtiter plate reader (Infnite F50 Wako, TECAN).Te percentage of bioflm inhibition was calculated by the formula: [(Control OD 620nm − Test OD 620nm ) ÷ Control OD 620nm ] × 100.Te amount of bioflm formed was measured by comparing the absorbance values of the extracttreated bacteria against the untreated ones.

Data Analyses.
All experiments and tests were performed in triplicates.All quantitative data were compared using relevant descriptive and inferential statistics using SPSS ver.20.0.Results of data analyses are given as mean of triplicates and standard deviation (mean ± SD) values, and

Results
Te specimens were identifed as Trametes elegans (from Arabuko-Sokoke National Forest) and Auricularia auriculajudae, Microporus xanthopus, Termitomyces umkowaani, and Trametes versicolor (from Kakamega National Forest) based on morphological characteristics and molecular techniques, respectively (Figure 1).Te specimens were identifed with the aid of mycologists and with careful observation of the physical characteristics of the mushrooms, spore prints, local knowledge, literature, taxonomy keys, and other sources.Te color and shape of the cap, the color and shape of the stipe, and the size and shape of the gill were some of the morphological characteristics used to identify mushrooms.Te majority of the specimens that were gathered have even gill margins, a centrally located stipe, equal stipe bases, and smooth pileal margins and surfaces.Te structure of the cap, the margin and placement of the gills, the location, surface, and shape of the stipe, and the margin, shape, and surface of the pileus, among other morphological observations of the mushrooms, were also noted (Table 1).Te ornamentation of the pileus and stipe surfaces, the presence or absence of an annulus on the stipe, and the presence or absence of a volva at the base of the stipe were additional characteristics that were used to defne and identify the mushrooms.However, none of the specimens had any volva or annulus features.Te only specimen without a stipe on its cap was Auricularia auricular-judae.Te color of the upper and lower surfaces of the caps of the gathered mushrooms was noted in addition to the aforementioned variables.
Te fve mushrooms' amplifed DNA bands have sizes ranging from 350 base pairs to 600 base pairs (Figure 1).Te ITS markers can clearly distinguish between and among species of mushrooms.Te examined mushrooms were identifed as A. auricula-judae, M. xanthopus, T. umkowaani, T. elegans, and T. versicolor.
At concentrations below the minimum inhibitory concentrations (if any), all A. auricula-judae extracts were evaluated for their antibacterial efectiveness against test microorganisms (Table 3).Te chloroform extract had antibacterial efects on E. coli.Extracts produced with hot water and 70% ethanol, however, exhibited no antibacterial action at the investigated concentration.
M. xanthopus extracts' percentage of bioflm-inhibitory activities was evaluated in a concentration-dependent manner (Figure 5).When compared to the control bacteria, the test bacteria signifcantly reduced the production of bioflms.Hot water and chloroform extracts reduced the bioflm-formation activity of P. aeruginosa (51.72 ± 0.01%) and E. coli (64.58 ± 0.03%), respectively.Te 70% ethanol extract had signifcant bioflm-inhibitory efects against E. coli and S. aureus (85.71 0.01%) but had little impact on P. aeruginosa (33.33 0.03%) in terms of bioflm formation.All of the extracts had minimal bioflm-inhibitory efects against P. aeruginosa.
Te efects of M. xanthopus extracts on the development of planktonic cells were assessed in comparison to the test bacterium.Only the planktonic cell of S. aureus was suppressed by all the extracts.However, P. aeruginosa and E. coli planktonic cells' proliferation has not been inhibited by the extracts (Table 4).

Bioflm-Inhibitory Activities of Termitomyces umkowaani
Extracts.At a concentration of 250 g/mL, T. umkowaani extracts showed bioflm-inhibitory efects against P. aeruginosa, E. coli, and S. aureus (Figure 6).Inhibitory bioflm activity against all of the test microorganisms was lowest in the chloroform extract.All extracts resulted in limited bioflm-inhibitory activities against P. aeruginosa and E. coli as compared to S. aureus.Chloroform, 70% ethanol, and hot water extracts showed statistically signifcant bioflm-inhibitory activities against S. aureus (p ≤ 0.05).
Te percentage of bioflm-inhibitory activities of T. umkowaani extracts was evaluated against the test bacteria (Figure 7).All extracts showed a weak percentage of antibioflm-formation activities against E. coli.Te best bioflm-inhibitory activities were observed with chloroform (64.71 ± 0.01%), 70% ethanol (68 ± 0.01%), and hot water (71.43 ± 0.02%) extracts against the S. aureus.Of the Evidence-Based Complementary and Alternative Medicine three extracts, the hot water extract exhibited a better percentage of bioflm-inhibitory activities against all the test bacteria.Te 70% ethanol and hot water extracts signifcantly limited (as much as 50%) the bioflm formation of P. aeruginosa.
Te chloroform extract resulted in growth inhibition of E. coli but not P. aeruginosa and S. aureus.Yet, the 70% ethanol and hot water extracts did not result in any antibacterial activities in all the test bacteria (Table 5).
Te percentage of bioflm-inhibitory activities of T. elegans extracts was very weak against all the test bacteria (Figure 9).All the extracts showed a very weak percentage of bioflm-inhibitory activities against P. aeruginosa and E. coli.Comparatively speaking, the chloroform extract yielded the best percentage of bioflm-inhibitory activity against S. aureus (69.75 ± 0.01%) as compared to the hot water (50 ± 0.02%) and 70% ethanol (41.18 ± 0.01%) extracts.
Te efects of the extracts of T. elegans on the antibacterial and the bioflm-inhibitory activities were determined against all the test bacteria (Table 6).Although the chloroform extract showed antibacterial activity against E. coli, there was no such efect on P. aeruginosa and S. aureus.On the other hand, the 70% ethanol and hot water extracts demonstrated limited bioflm-inhibitory activities against all the test bacteria without any efect on their growth.

Bioflm-Inhibitory Activities of Trametes versicolor
Extracts.Te bioflm-inhibitory activities of the extracts of T. versicolor (250 μg/mL) against test and control bacteria yielded no statistically signifcant results (p > 0.05) (Figure 10).All the extracts resulted in a weak percentage of bioflm-inhibitory activities against all the test bacteria.
Te percentage of bioflm-inhibitory activities of T. versicolor extracts against the test microorganisms was investigated (Figure 11).A very small percentage of bioflminhibitory activity against P. aeruginosa and E. coli was produced by all three extracts.However, all extracts showed the highest percentage of bioflm-inhibitory activities against S. aureus.Hot water (41.18 ± 0.01%) and 70% ethanol (42.86 ± 0.01%) extracts exhibited the lowest and the highest percentage of bioflm-inhibitory activities against S. aureus.Evidence-Based Complementary and Alternative Medicine Besides, the 70% ethanol extract showed the highest percentage of bioflm-inhibitory activity against S. aureus (42.86 ± 0.01%) and lowest against E. coli (10.53 ± 0.01%).P. aeruginosa and S. aureus planktonic cells did not experience any growth suppression from the chloroform extract, despite the fact that E. coli did (Table 7).Both a 70% ethanol extract and a hot water extract had a discernible inhibitory efect on the growth of planktonic cells when used against S. aureus and P. aeruginosa.

Discussion
Te results of the current investigation demonstrated that A. auricula-judae and M. xanthopus extract in chloroform, 70% ethanol, and hot water had efective bioflm-inhibitory properties.Te ability of the extracts to inhibit the bioflmformation sites (such as cyclic diguanylate) of the test bacteria may be one of the many potential explanations for these promising bioflm-inhibitory capabilities.Cyclic diguanylate starts the synthesis of bis-(3′-5′)-cyclic dimeric guanosine monophosphate, which makes it possible to produce sticky compounds connected to the bioflm development process continuously [34].Te extracts may also be able to considerably repress a number of genes and prevent the production of sticky compounds necessary for the processes of quorum sensing and bioflm formation [35,36].
Te test bacteria's ability to form bioflms was signifcantly reduced by the bioflm-inhibitory efects of A. auricula-judae and M. xanthous extracts.However, the extracts displayed a range of bioflm-formation inhibitory efects against E. coli, P. aeruginosa, and S. aureus.P. aeruginosa's capacity to form bioflms was diminished by hot water extracts of A. auricula-judae and M. xanthopus.Similarly, all of the A. auricula-judae and M. xanthopus extracts showed superior bioflm-inhibitory properties against S. aureus but were less efcient in preventing P. aeruginosa from forming bioflms.Tis might be attributed to and explained by the two bacteria's contrasting efux pumps for the extracts and their distinct afnities for certain signaling molecules [37].Moreover, the recovered bioactive chemicals cannot enter the bacterial cells because of the structure of the polysaccharides that surround them [38].
Extracts of A. auricula-judae and M. xanthous prevented the test bacteria from forming bioflms without impairing the growth of their planktonic cells.Tese results show that the extracts prevented or reduced the test bacteria's ability to form bioflms at a concentration of 50 g/mL, which is much lower than the minimum inhibitory concentration values noted in our earlier studies [39,40].Te receptor proteins and molecules involved in the quorum-sensing pathway may have had their activity reduced by these extracts.According certain research, extract concentrations below MIC values do not necessarily kill the test bacteria; instead, they have an alternative method of action that prevents the adhesion process [31].Yet, this is not always the case.An investigation into oral bioflms revealed that extracts of Lentinula edodes at and below the MIC values did not signifcantly slow down the bioflm-formation process [41].
Even though all of the A. auricula-judae and M. xanthopus extracts only had modest antimicrobial efects on P. aeruginosa, they had signifcant bioflm-inhibitory efects.According to these results, extracts with good   Evidence-Based Complementary and Alternative Medicine antimicrobial activity do not necessarily have good bioflm suppressive activities [42].Tis may be explained by the fact that bioflms have unique phenotypic characteristics that are not present in their planktonic cousins.For example, bioflm-forming bacteria boost rates of genetic exchange among other bioflm occupants and boost resistance to environmental variables such antibiotics, detergents, UV exposure, dehydration, salt, and phagocytosis [42][43][44].
Te bioflm-forming ability of S. aureus was severely reduced by all T. umkowaani extracts.Tese extracts' capacity to prevent the development of the enzymes and         Evidence-Based Complementary and Alternative Medicine proteins necessary for the creation of extracellular polymeric compounds may explain their ability to suppress bioflminhibitory efects against S. aureus.Tese results are consistent with those of earlier studies that looked at the antibioflm properties of aqueous extracts against Streptococcus mutans [45].Tis may be because these extracts inhibit DNA gyrase, which is necessary for DNA synthesis.Similar to this, the extracts' abilities to suppress bioflm formation could be brought on by the bacterium's blockage of quorum sensing.Extracts' antiquorum-sensing abilities have the ability to quickly alter the gene expression patterns of a bacterial population in response to population density [31].S. aureus's ability to form bioflms was markedly reduced by a hot water extract of T. elegans.Tis may be related to the extracts' detrimental efects on the bacterial motility and extracellular polymer synthesis during the bioflm development stages [46].Additionally, the extracts may be able to obstruct the early phases of bioflm development by preventing the production of extracellular polysaccharide molecules and the initial adhesion of the bacterial cells to the surfaces [47][48][49][50][51].
Comparatively to the standard bacterial strain of S. aureus, all T. versicolor extracts showed poor bioflminhibitory efects against the clinical isolates of P. aeruginosa and E. coli.Te observed weak antibioflm efects of the extracts on the clinical isolates, inherent resistance to antimicrobial agents and some genetically controlled processes, may be responsible for the observed weak bioflminhibitory activities [52,53].Strong bioflm development, as seen in the P. aeruginosa isolates, is a crucial component in the pathogenesis of many bacterial illnesses as well as their survival and fourishing in a variety of habitats.In order to establish a host, increase their population, and, most critically, spread disease, bacterial pathogens may rely on the emergence of bioflms [52,54,55].

Conclusions
Te results suggest that the mushroom extracts would be a good option for bioflm-inhibitory properties.In a concentration-dependent pattern, all mushroom extracts demonstrated considerable bioflm-inhibitory efcacy against the test organisms.All extracts were more efcient against S. aureus at inhibiting bioflms.Te intriguing potential of the extracts to inhibit the bacteria's bioflmformation processes might lead to the discovery of new chemicals that might fnd use in clinical settings.Researchers may be prompted to hunt for new substances with potential for use in medicine by the extracts' promising capacity to signifcantly block the test bacteria's bioflm-formation activities.To pinpoint the active extract ingredients that are responsible for the extracts' bioflm-inhibiting properties, more research is required.It is vital to identify these bioactive substances and explain how they work in order to exert their antibioflm efects.

Future Prospective of the Study.
Te pharmaceutical and medical biotechnology sectors, as well as human health, will be signifcantly impacted by the research fndings discussed in this paper.Terefore, it is crucial to utilize wild

Figure 3 :
Figure 3: Percentage of bioflm-inhibitory activities of A. auriculajudae extracts against the test bacteria.

Figure 5 :
Figure 5: Percentage of bioflm-inhibitory activities of M. xanthopus extracts against the test bacteria.

Figure 7 :
Figure 7: Percentage of bioflm-inhibitory activities of T. umkowaani extracts against the test bacteria.

Figure 8 :
Figure 8: Bioflm-inhibitory activities of T. elegans extracts against the test bacteria.

Figure 9 :
Figure 9: Percentage of bioflm-inhibitory activities of T. elegans extracts against the test bacteria.

Figure 10 :
Figure 10: Bioflm-inhibitory activities of T. versicolor extracts against the test bacteria.

Figure 11 :
Figure 11: Percentage of bioflm-inhibitory activities of T. versicolor extracts against the test bacteria.

Table 1 :
Morphological characters and keys used for the identifcation of the wild mushrooms.
°C.Te extracts were kept in a −80 °C deep freezer and freeze-dried (MRC Freeze Dryer, Model, FDL-10N-50-8M).Finally, crude extracts were stored in a 4 °C refrigerator in amber-colored bottles for further analyses.

Table 2 :
Primers used to amplify specifc genes of the fve mushroom species.

Table 3 :
Efect of A. auricula-judae extracts on the growth of planktonic bacterial cells.: negative control; all extracts were tested at 62.5 μg/mL.Te signifcance of the bold values is to indicate that the chloroform extract has shown promising results beyond the negative control. NC

Table 4 :
Efect of M. xanthopus extracts on the growth of planktonic bacterial cells.NC: negative control; all extracts were tested at 250 μg/mL.Te signifcance of the bold values is to indicate that all the extracts have shown promising results against S. aureus even beyond the negative control.

Table 5 :
Efect of T. umkowaani extracts on the growth of planktonic bacterial cells.

Table 6 :
Efect of T. elegans extracts on the growth of planktonic bacterial cells.Te signifcance of the bold values is to indicate that the chloroform extract has shown promising results against E. coli even beyond the negative control.
NC: negative control; all extracts were tested at 250 μg/mL.

Table 7 :
10ect of T. versicolor extracts on the growth of planktonic bacterial cells.Te signifcance of the bold values is to indicate that all extracts have shown promising results against all bacteria even beyond the negative control.10Evidence-BasedComplementary and Alternative Medicine mushrooms as food and a unique source of antibacterial and antibioflm chemicals.With the aid of these bioactive molecules, the search for novel compounds to combat the troublesome characteristics of bioflm-forming and drugresistant bacteria might also enter a new era.
NC: negative control; all extracts were tested at 250 μg/mL.