Modern approaches for detection of volatile organic compounds in metabolic studies focusing on pathogenic bacteria:Current state of the art

Pathogenic microorganisms produce numerous metabolites,including volatile organic compounds(VOCs).Monitoring these metabolites in biological matrices(e.g.,urine,blood,or breath)can reveal the presence of specific microorganisms,enabling the early diagnosis of infections and the timely implementation of tar-geted therapy.However,complex matrices only contain trace levels of VOCs,and their constituent com-ponents can hinder determination of these compounds.Therefore,modern analytical techniques enabling the non-invasive identification and precise quantification of microbial VOCs are needed.In this paper,we discuss bacterial VOC analysis under in vitro conditions,in animal models and disease diagnosis in humans,including techniques for offline and online analysis in clinical settings.We also consider the advantages and limitations of novel microextraction techniques used to prepare biological samples for VOC analysis,in addition to reviewing current clinical studies on bacterial volatilomes that address inter-species in-teractions,the kinetics of VOC metabolism,and species-and drug-resistance specificity.

Antiseptic Efficacy of A Soap Made from Biosurfactants Isolated from Bacillus and Lactobacillus against Pathogenic Bacteria

The aim of our study was to use a biosurfactant produced by Bacillus and Lactobacillus isolates as an antiseptic in the formulation of local soap. A total of 60 isolates were characterized by microbiological techniques (30 Bacillus and 30 Lactobacillus) and the ability to produce biosurfactants was demonstrated by a hydrocarbon emulsification index (E24). The emulsification indexes (E24) varied from 9% to 100% for Bacillus and from 33% to 100% for Lactobacillus as well. The antagonistic assay showed that biosurfactants were able to inhibit the formation of biofilms and growth of pathogens such as Escherichia coli, Pseudomonas aeruginosa, Staphylococcus aureus, Bacillus cereus, Salmonella typhirium, Shigella boydii and Proteus mirabilis. The biosurfactant consortium (BioC) from Bacillus consortium and from Lactobacillus was able to inhibit biofilm formation and the pathogens growth. The BioC was stable to alkaline pH and the temperatures stability of Biosurfactant was ranging from 50°C to 90°C. The soap was made by the cold saponification process using one biosurfactant consortium formulated. This soap has a pH of 10 and showed good cleaning power and good foam stability. Similarly, the soap showed good antiseptic power and disinfection power against all pathogens tested. Handwashing is critical to preventing disease transmission. The persistence of pathogens in waste water was evaluated. The BioS produced showed good disinfection power against all pathogens tested. The valor of reduction on the hands and in the waste water was significantly more than compared to the control soaps used. This soap could be used in the prevention, fighting, and treatment of bacterial and viral infections.

Distribution of pathogenic bacteria and antimicrobial sensitivity of eye infections in Suzhou

AIM:To investigate the types of bacteria in patients with eye infections in Suzhou and their drug resistance to commonly used antibacterial drugs.METHODS:The clinical data of 155 patients were retrospectively collected in this study,and the pathogenic bacteria species and drug resistance of each pathogenic bacteria were analyzed.RESULTS:Among the 155 patients(age from 12 to 87 years old,with an average age of 57,99 males and 56 females)with eye infections(160 eyes:74 in the left eye,76 in the right eye and 5 in both eyes,all of which were exogenous),71(45.81%)strains were gram-positive bacteria,23(14.84%)strains were gram-negative bacteria and 61(39.35%)strains were fungi.Gram-positive bacteria were highly resistant to penicillin and erythromycin(78.87%and 46.48%respectively),but least resistant to vancomycin at 0.Gram-negative bacteria were highly resistant to cefoxitin and compound sulfamethoxazole(100%and 95.65%respectively),but least resistant to meropenem at 0.Comparison of the resistance of gram-positive and gram-negative bacteria to some drugs revealed statistically significant differences(P<0.05)in the resistance of both to cefoxitin,cotrimoxazole,levofloxacin,cefuroxime,ceftriaxone and ceftazidime,and both had higher rates of resistance to gram-negative bacteria than to gram-positive bacteria.The distribution of bacterial infection strains showed that Staphylococcus epidermidis was the most common strain in the conjunctiva,cornea,aqueous humor or vitreous body and other eye parts.Besides,Fusarium and Pseudomonas aeruginosa were also among the most common strains of conjunctival and corneal infections.CONCLUSION:Gram-positive bacteria are the dominant bacteria in eye infections,followed by gram-negative bacteria and fungi.Considering the resistance of gramnegative bacteria to multiple drugs,monitoring of bacteria should be strengthened in eye bacterial infections for effective prevention and control to reduce complications caused by eye infections.

A Natural Catalytic Converter® for Continuously Inactivating Air and Surface Pathogens with More Effect than Ventilation and Filtration

Study Objective: The purpose of the study is to present independent laboratory testing for a novel technology in air and on surfaces. Since 2020, public health goals have focused on improving indoor air quality. This includes protection from airborne pathogens, such as tuberculosis, RSV, SARS-CoV-2, common cold or influenza viruses, measles, and others. Engineering controls are highly effective at reducing hazardous pathogens found in indoor air and from recontamination of surfaces. This occurs from a continuous cycle of settling of small, sustained airborne pathogens, which may become dehumidified, becoming airborne again, carried by room air currents around indoor spaces, then repeating the cycle. Methods: The novel technology utilizes a catalytic process to produce safe levels of hydrogen peroxide gas that are effective in reducing pathogens in the air and on surfaces. Air testing was performed with the MS2 bacteriophage, the test organism for ASHRAE standard 241, and methicillin-Resistant Staphylococcus aureus (MRSA). Surface testing was performed with SARS-COV-2 (Coronavirus COVID-19) and H1N1 (Influenza). Typical ventilation and filtration does not effectively remove disbursed pathogens from the entire facility, due to inconsistent air circulation and surface deposits of pathogens. Results: MS2 was reduced by 99.9%;MRSA was reduced by 99.9%;SARS-CoV-2 was reduced by 99.9%;H1N1 was reduced by 99.9%. Conclusion: This novel catalytic converter reduces a variety of pathogens in the air (99%) and on surfaces (99%), by actively disinfecting with the introduction of gaseous hydrogen peroxide. This active disinfection provides a strong solution for protecting the entire facility and its occupants.

Bio-control of Some Food-Borne Pathogenic Bacteria by Bacteriophage

In recent years, researchers tended to bring new alternative to biological protective systems used in conservation of food and production of safe food. Use of bacteriophage against to pathogen bacteria in food was the most hopeful system in these methods about bio-control. Controls of bacteriophage for each pathogen species and subspecies and determination of phage-host originality are important because efficient bio-control was achieved. Researches concentrated on some food-borne pathogen bacteria such as E. coli O157：H7, Campylobacter, Salmonella and Listeria. In a consequence of these studies made as in vitro and in vivo, first commercial production of phage which will be used in foods was made in Netherlands. Also, it has been informed that use of phage is cost-efficient alternative as compared with other preservatives. This review, discussed application of bacteriophages as bio-control agents in food and advantages and disadvantages about uses of bacteriophages by taking into account antimicrobial characteristics of them.

Pediococcus Acidilactici Inhibit Biofilm Formation of Food-Borne Pathogens on Abiotic Surfaces

In this study, we aimed to examine the inhibitory effect of PA003, a Pediococcus acidilactici that produces lactic acid and antimicrobial peptides pediocin, on pathogenic biofilm formation on abiotic surfaces. PA003 and pathogens (Escherichia coli, Salmonella enterica serovar Typhimurium, Staphylococcus aureus and Listeria monocytogenes) were used to evaluate auto-aggregation, hydrophobicity, biofilm formation and biofilm formation inhibition on stainless steel, polyvinyl chloride and glass slides in terms of exclusion, displacement and competition. The results showed the highest auto-aggregation abilities were observed for one of the E. coli strains EAggEC (E58595) and the highest hydrophobic strain was observed with EPEC (E2348/69) (51.9%). The numbers of biofilm cells of E. coli, S. Typhimurium, S. aureus and L. monocytogenes on stainless steel, polyvinyl chloride and glass slide coupons were effectively reduced by approximately 4 log CFU/coupon. These results demonstrate that lactic acid bacteria can be used as an alternative to effectively control the formation of biofilms by food-borne pathogens. © 2016, Tianjin University and Springer-Verlag Berlin Heidelberg.

Diversity of Gram negative bacteria antagonistic against major pathogens of rice from rice seed in the tropic environment

With the use of a seed washing technique, more than 4000 Gram negative bacteria were isolated by two improved isolation methods from 446 batches of 1 kg rice seed samples obtained from 22 provinces in the Philippines. They were initially characterized on the basis of colony morphology and results of biochemical and pathogenicity tests. Six hundred and fifty two strains were further identified by Biolog, from which 133 were selected for fatty acid methyl ester (FAME) analysis together with 80 standard reference!strains. Sixteen species or types of Pseudomonas and 17 genera of non pseudomonads were identified, more than one third of which have not been recorded in rice. The most predominant species observed were P. putida and P. fulva. About 17% of the strains of Pseudomonas and 2% of the non pseudomonads were antagonistic to one or more fungal or bacterial pathogens of rice. Rice seed is an important source of biological control agents.

In vitro and in vivo bactericidal activity of Vitex negundo leaf extract against diverse multidrug resistant enteric bacterial pathogens

Objective:To investigate in vitro and in vivo antibacterial potentials of Vitex negundo(V. negundo) leaf extracts against diverse enteric pathogens.Methods:Water and methanol extracts of V.negundo leaves were evaluated against enteric bacterial pathogens by using standard disc diffusion,viable bacterial cell count methods,determination of minimum inhibitory concentrations(MIC) and minimum bactericidal concentrations(MBC).Results:Methanol extract of V.negundo leaves showed potent antibacterial activity(inhibition zone:9.9-22.6 mm,MIC: 200-3 200μg/mL.MBC:200-6 400μg/mL) against all the pathogenic enteric bacteria(Vibrio cholerae.Vibrio parahemolyticus.Vibrio mimicus.Echerichia coli,Shigella spps.,and Aeromonas spps) tested.Methanol extract of V.negundo leaves showed potent bactericidal activity both in vitro laboratory conditions(MBC,200-400μg/mL) and in the intestinal environment(Dose,1-2 mg/mL) of infant mice against pathogenic Vibrio cholerae,the major causative agent of cholera. Furthermore,assays using the mice cholera model showed that V.negundo methanol extract can protect mice from Vibrio cholerae infection and significandy decrease the mortality rate(P【0.0001). Conclusions:For the first time we showed thal medianol extract of V.negundo leaves exhibited strong vihriocidal activity both in vitro and in vivo conditions.Therefore,it will he useful to identify and isolate the active compounds of this extract that could be a good alternative of antibiotics to treat cholera.

Antimicrobial Profile of Lactic Acid Bacteria Isolated from Vegetables and Indigenous Fermented Foods of India against Clinical Pathogens Using Microdilution Method

In dairy and food industries lactic acid bacteria （LAB） have been used in form of starter culture that plays vital role in fermentation; as flavouring and texturizing or as preservative agents. There is increasing evidence that lactobacilli which inhabit the gastrointestinal tract develop antimicrobial activities and participate in the host＇s defence system[1]. During fermentation, most of the LAB produces a number of different compounds like organic acids, hydrogen peroxide, diacetyl, acetaldehyde, carbon dioxide, polysaccharides, and proteinaceous compounds called bacteriocins or bacteriocinogenic peptides.

In vitro and in vivo bactericidal activity of Vitex negundo leaf extract against diverse multidrug resistant enteric bacterial pathogens

Objective:To investigate in vitro and in vivo antibacterial potentials of Vitex negundo(V. negundo) leaf extracts against diverse enteric pathogens.Methods:Water and methanol extracts of V.negundo leaves were evaluated against enteric bacterial pathogens by using standard disc diffusion,viable bacterial cell count methods,determination of minimum inhibitory concentrations(MIC) and minimum bactericidal concentrations(MBC).Results:Methanol extract of V.negundo leaves showed potent antibacterial activity(inhibition zone:9.9- 22.6 mm,MIC: 200-3 200μg/mL,MBC:200-6 400μg/mL) against all the pathogenic enleric bacteria(Vibrio cholerae,Vibrio parahaemolyticus,Vibrio mimicus,Echerichia coli.Shigella spps.,and Aeromonas spps) tested.Methanol extract of V.negundo leaves showed potent bactericidal activity both in vitro laboratory conditions(MBC,200-400μg/mL) and in the intestinal environment(Dose,1-2 mg/mL) of infant mice against pathogenic Vibrio cholerae,the major causative agent of cholera. Furthermore,assays using the mice cholera model showed that V.negundo methanol extract can protect mice from Vibrio cholerae infection and significantly decrease the mortality rate(P【0.0001).Conclusioas:For the first time we showed that methanol extract of V.negundo leaves exhibited strong vibriocidal activity both in vitro and in vivo conditions.Therefore,it will be useful to identify and isolate the active compounds of this extract that could be a good alternative of antibiotics to treat cholera.

Resistance of Microbial Community of Artemisia annua L.to Pathogenic Fungi

[Objectives]This paper was to figure out whether the dominant bacterial community has the role and effect of bacterial community and its defense mechanism against potential pathogenic fungi in Artemisia annua,and thus establish a systematic model of bacteria-fungus-plant.[Methods]Fifty-eight strains of bacteria and one strain of pathogenic fungi,Globisporangium ultimatum,were used for the experiments.These 58 bacterial strains were assembled into a bacterial community,and the bacteria with abundance in the top 1%were reassembled into a dominant bacterial community as measured by 16S rDNA.[Results]The growth of A.annua seedlings inoculated with bacterial communities and pathogenic fungi or dominant bacterial communities and pathogenic fungi was significantly better than that of A.annua seedlings inoculated with pathogenic fungi during in vitro confrontation,which was evident in both enzymatic and non-enzymatic antioxidant assays.[Conclusions]The results suggest that the dominant bacterial community has a crucial role as a representative core microbial community of synthetic bacterial community,which can protect plants by interfering with the growth of phytopathogenic fungi mediated by chemical signals,and can be used as the main synthetic community of biocides to achieve the effect of biocontrol.

Antimicrobial Activity of Wild Plant Seed Extracts against Human Bacterial and Plant Fungal Pathogens

Five wild plant species belonging to different families (Chenopodium album, Plantago major, Elytrigia elongata, Filipendula ulmaria and Nigella sativa) widely spread in Russian Federation and the former USSR were evaluated for their ability to inhibit growth of two important human food-borne pathogens (Escherichia coli O157:H7 and Listeria monocytogenes strain EGD-e) and eight plant pathogens (Alternaria alternata, Alternaria tenuissima, Bipolaris sorokiniana, Stagonospora nodorum, Fusarium solani, Fusarium oxysporum, Fusarium culmorum and Phytophtora infestans). To isolate biologically active compounds from seeds, a step-wise procedure including extraction with hexane, ethyl acetate, ethanol, and 10% acetic acid followed by reversed-phase HPLC was developed. Using disc-diffusion assay, the highest activity against E. coli O157:H7 was observed with extracts from F. ulmaria (hexane and ethyl acetate extracts and the unbound RP-HPLC fraction) and P. major (ethyl acetate extract and the unbound RP-HPLC fraction);E. elongate (the unbound RP-HPLC fraction) was less active. The extracts from P. major and E. elongate (the unbound RP-HPLC fractions) were equally highly active against L. monocytogenes, while those of F. ulmaria (the unbound RP-HPLC fraction) and N. sativa (hexane and ethyl acetate extracts) were less active against this pathogen. The dynamics of L. monocytogenes EGD-е and E. coli O157:H7 growth in the presence of two most potent extracts (RP-HPLC-unbound fractions of P. major and E. elongate and the hexane extract of F. ulmaria) was studied.

Sanger Sequencing Implementation in Clinically Ill Patients for Bacterial and Fungal Pathogens Identification

The diagnosis of bacterial or fungal infections requires the identification of the pathogen etiology in the shortest time possible. Although some biomarkers are used as indicators of bacterial infections, their specificity and sensitivity are highly variable, and there is no direct relationship between the level increase of these biomarkers for mycosis. It is common to obtain negative microbiological cultures in patients infected by non-culturable, intracellular bacteria or mycosis, even though there is a high clinical suspicion of infection. This study identifies the pathogen present in critically infected patients through 16S and 18S/eEF1 genes detection by polymerase chain reaction (PCR) coupled with Sanger sequencing. Thirty clinical samples were evaluated by PCR, of which 40% were positive for fungi, 23.33% for bacteria, 26.7% for fungi and bacteria, and 10% for no pathogen. The PCRs outcomes period for bacteria or fungi was one day compared to seven and up to 14 days (on average) of microbiological culture for bacteria and fungi. Then, we assessed the relationship with the most used biomarkers (procalcitonin, C-reactive protein, globular sedimentation velocity, and the neutrophil-lymphocyte index). This combination of molecular techniques has been shown as helpful in identifying intracellular bacteria and fungi that are difficult to culture by conventional methods. Screening with genomic markers 16S and 18S/eEF1 by PCR allowed us to optimize the time to obtain the result of the infection caused by bacteria or fungi. Also, identifying the specific etiological microorganism by Sanger sequencing was very helpful in avoiding the progression of the disease and setting targeted treatment with better clinical outcomes.

STUDY ON ADHERENCE ABILITY OF PERIODONTAL PATHOGENS AND CARIOGENIC BACTERIA TO HYDROXYAPATITE DISKS

Objective To study the adherence activity of six representative periodontopathic and cariogenic bacteria to hydroxyapatite disks. Methods Six periodontopathic and cariogenic bacteria of P. gingivalis, A. actinomycetemcomitans, F. nucleatum, S. sanguis, A. viscosus and S. mutans were cultured in modified MD-300 chemostat according to total fifteen experimental groups of single-specie and each pair of periodontal pathogens and cariogenic bacteria, respectively. After 1h attached live bacteria on removable hydroxyapatite disks was analyzed by culture technologies to evaluate the adherence level. Results The adherence activity of periodontopathic and cariogenic bacteria to HA was in the following order: S. sanguis > A. viscosus> S. mutans> A. actinomycetemcomitans > F. nucleatum > P. gingivalis. The number of periodontopathic bacteria to HA was enhanced by S. sanguis and A. viscosus,respectively.When mix-cultivated with S. mutans, the colonization of P. gingivalis was reduced significantly(P<0.001). Periodontopathic bacteria had no effect on the adherence activity of S. mutans and A. viscosus, except S. sanguis. Conclusion It was showed that the adherence activity of periodontal pathogens was weaker than that of cariogenic bacteria and emphasized the importance of bacterial adherence in determining the level of bacterial colonization on tooth surfaces. It was suggested that periodontopathic bacteria can utilize initial colonizers to become those predominant bacteria in periodontal ecosystem, which maybe have close relation to the periodontopathic mechanism.

Drug Sensitive Test on Main Pathogenic Bacteria of Cow Recessiveness Mastitis Using Traditional Chinese Medicine Preparation “Zengrujianniusan”

According to the theory of Chinese veterinary medicine and characteristics of Chinese herbal medicine, four different preparations of compound Chinese medicine ＂Zengrujianniusan＂ were composed, and the prepared water decoction was used to carry on the bacteriostatic test on main pathogens of cow recessiveness mastitis. The results showed that the four different prescriptions of water decoction all had antibacterial effects. The prescription 3 was sensitive to Staphylococcus aureus and Streptococcus agalactiae, while the other three prescriptions showed high sensitivity, and the prescription 3 had the strongest bacteriostatic effects.

Antibacterial efficacy of the seed extracts of Melia azedarach against some hospital isolated human pathogenic bacterial strains

Objective:To investigate the antibacterial potential of the polar and non-polar extracts of the seeds of Melia azedarach(M.azedarach) L.(Meliaceae) against eighteen hospital isolated human pathogenic bacterial strains.Methods:Petrol,benzene,ethyl acetate,methanol,and aqueous extracts at five different concentrations(1,2,5,10 and 15 mg/mL) were evaluated.Disk diffusion method was followed to evaluate the antibacterial efficacy.Results:All extracts of the seeds demonstrated significant antibacterial activity against tested pathogens.Among all extracts,ethyl acetate extract revealed the highest inhibition comparatively.The present study also favored the traditional uses reported earlier.Conclusions:Results of this study strongly confirm that the seed extracts of M.azedarach could be effective antibiotics,both in controlling gram-positive and gram-negative human pathogenic infections.

Survival of pathogenic bacteria in compost with special reference to Escherichia coli

Application of compost in agricultural practice could potentially cause contamination of foodstuffs with pathogenic bacteria such as Escherichia coli O157：H7 （E. Coli O157）. We investigated pathogenic bacteria in compost collected from the compost facilities, and evaluated the survival of E. coli K12 and O157 in laboratory experiments. Out of 19 compost product samples, coliform bacteria and salmonella were detected in 7 and 3 samples respectively. The number of coliform bacteria was 1.8 × 10^2 to 2.5 × 10^6 CFU/g dw and that of salmonella was 4.2 × 10^1 to 6.0 × 10^3 CFU/g dw. Moreover. coliform bacteria, fecal coliform, E. coli and salmonella were detected during composting at 54℃ to 67℃. The results indicated that moisture content was a very important factor to the heat sensitivity of pathogenic bacteria in compost, E. coli in compost of high moisture content was more sensitive than that in compost of low moisture content, cells harvested in logarithmic phase was more sensitive than these in stationary phase, and E. coli K12 was more sensitive than E. coli O157. Based on the D values, the lethal time of E. coli K12 and 0157 from 10^8 to 10^0 CFU/g dw were 16.3 and 28.8 min, respectively, at 60℃ in compost with 40% moisture content. However, some E. coli cells survived in composting process at 54℃ to 67℃. Water potential（low moisture content） and physiological aspects of bacteria（stationary phase） could explain only in part of the prolonged survival of E. coli in compost, and there should be some other factors that are conducive to bacterial survial in compost.

Fucoxanthin Isolated from Undaria pinnatifida Can Interact with Escherichia coli and lactobacilli in the Intestine and Inhibit the Growth of Pathogenic Bacteria

Fucoxanthin is a xanthophyll-type carotenoid that provides many benefits to human health. However, the mechanism by which fucoxanthin interacts with microbes and inhibits pathogenic bacteria is unknown. In this study, we investigated the effects of fucoxanthin isolated from the edible seaweed Undaria pinnatifida on pathogenic bacteria Escherichia coli and lactobacilli both in vitro and in vivo. Fucoxanthin strongly inhibited the growth of Gram-positive pathogenic bacteria but was less effective against Gram-negative bacteria. Fucoxanthin extracted from the crude mixture had a recovery rate of 93.38% and a purity of 82.70%, which were higher than those of fucoxanthin extracted using a previous method. Fucoxanthin also promoted the growth of intestinal mi- crobes in mice. Fucoxanthinol, a metabolite of fucoxanthin, was generated in the culture media. Fucoxanthin can be deacetylated into fucoxanthinol not only by conventional digestive enzymes in the digestive tract, but also by E. coli and lactobacilli in the intestine. These results indicate that fucoxanthin interacts with and influences E. coli and lactobacilli in the intestine. Therefore, fucoxanthin isolated from Undaria pinnatifida possibly can be applied in human health maintenance.

In vitro antibacterial potential of some Vitex species against human pathogenic bacteria

Objective:To study the antibacterial activity of the leaf methanol extracts of five different species of Vitex namely,Vilex altissima(V.allissima),Vitex diversifolia(V.diversifolia),Vitex negundo(V.negundo),Vitex peduncularis(V.peduncularis) and Vitex trifolia(V.trifolia). Methods:Antibacterial assay was carried out by using disc diffusion method,determination of minimum inhibitory concentrations(MIC) and minimum bactericidal concentrations(MBC) against five strains of Gram-positive and seven strains of Gram-negative human pathogenic bacterial strains.Results:The results of antibacterial activity of Vitex species showed that the extracts possessed a broad spectrum of antibacterial activity.The V.peduncularis possessed the highest activity against all the microorganisms screened.It produced a zone of inhibition ranged between (11.000±0.577) and(22.670±0.667) mm and the MIC values were from 62.5 to I 000.0μg/mL and the MBC values were from 125.0 to 2 000.0μg/mL.Conclusions:Based on the present study, V.peduncularis is recommended for the isolation of antibacterial molecule responsible for the activity against the tested human pathogenic bacterial strains.

Antibacterial activity of leaves extracts of Trifolium alexandrinum Linn,against pathogenic bacteria causing tropical diseases

Objective:To investigate antibacterial potential of Trifolium alexandrinum(T.alexandrinum) Linn,against seven gram positive and eleven gram negative hospital isolated human pathogenic bacterial strains responsible for many tropical diseases.Methods:Non-polar and polar extracts of the leaves of T.alexandrinum i.e.,hexane,dichloromethane(DCM),ethyl acetate(EtOAc), methanol(MeOH) and aqueous(AQ) extracts at five different concentrations(1,2,5,10 and 15 mg/ mL) were prepared to evaluate their antibacterial value.NCCL standards were strictly followed to perform antimicrobial disc susceptibility test using disc diffusion method.Results:Polar extracts demonstrated significant antibacterial activity against tested pathogens.EtOAc and MeOH extracts showed maximum antibacterial activity with higher inhibition zone and were found effective against seventeen of the tested pathogens.While AQ plant extract inhibited the growth of sixteen of the test strains.EtOAc and MeOH plant extracts inhibited the growth of all seven gram positive and ten of the gram negative bacterial strains.Conclusions:The present study strongly confirms the effectiveness of crude leaves extracts against tested human pathogenic bacterial strains causing several tropical diseases.Since Egyptian clover is used as a fodder plant,it could be helpful in controlling various infectious diseases associated with cattle as well.