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.

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.

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.

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.

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.

Isolation and Identification of Houttuynia cordata Endophytic Bacteria and Study of Antibiotic Activity

[Objectives]To further explore the resources of biocontrol strains and provide new options for the control of pathogenic bacteria.[Methods]Houttuynia cordata endophytic bacteria were isolated and identified,and the antibiotic activity was explored.[Results]There are 8 strains(H1-H8)of endophytic bacteria isolated from the healthy plants of H.cordata,which had an antagonistic effect on the pathogenic bacteria.According to the gene sequence,the 8 strains were identified as Stenotrophomonas,Pseudomonas,and Bacillus,which strongly inhibited Colletotrichum(HC1),Fusarium(HC2),and Rhizoctonia(HC5)of H.cordata.On the PDA culture dish,the inhibition diameter of the 8 strains on Colletotrichum,Fusarium,and Rhizoctonia was 6,8,and 5 mm,respectively.[Conclusions]Pathogenic bacteria and endophytic bacteria inoculated with H.cordata seedlings showed that endophytic bacteria have certain antagonistic activity,and have the potential for development and application.

Ag enhanced CuS nanoflower catalyst coupling dielectric barrier discharge plasma for disinfection performance and mechanism

In this study,the hydrothermal method was employed to grow submicron CuS on carbon cloth(CC),and the photoreduction method was used to grow Ag nanoparticles on the CuS submicron flowers,thus forming the Ag/CuS/CC catalytic electrode.The application of Ag/CuS/CC electrode-coupled dielectric barrier discharge(DBD)plasma in the disinfection of pathogenic bacteria in water was studied.The Ag/CuS/CC electrode exhibits strong antibacterial activity,and under an external voltage of 30 V,the degradation efficiency of Bacillus subtilis reaches 99.99%within 15 min without regeneration.After five cycles,the inactivation rate of Bacillus subtilis reached 99.99%within 25 min.The practical applicability of the Ag/CuS/CC-coupled DBD system for treating actual wastewater was evaluated,and the changes in biological toxicity were investigated.The results indicate that the prepared Ag/CuS/CC coupled DBD has great potential for safe disinfection of pathogenic bacteria in water through integrated processes.

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.

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.

Studies on the edwardsiellosis and characterization of pathogenic bacteria from diseased flounder (Paralichthys olivaceus L.) and turbot (Scophthalmus maximus L.)

Edwardsiellosis of flounder and turbot occurring in different mariculture farms during 2001～ 2004 was examined, including the conditions of disease occurrence, clinical signs and pathological changes. The results showed that all diseased fishes expressed bacterial septicaemia. A total of 148 strains were identified using a combination of traditional physiological and biochemical tests and partial 16S rRNA gene analysis. In addition, the mole fraction G ＋ C ratio of the DNA of representative strain of isolates and serum homology were detected, and pathogenicity tests of isolates were conducted by experimental infection. The results revealed that 148 strains were identified as E. tarda of genus Edwardsiella, all the isolates are of serologic similarity, and have strong pathogenicity to flounder and turbot.

Distribution and drug resistance of pathogenic bacteria isolated from cancer hospital in 2013

Objective： To understand distribution and drug resistance of pathogenic bacteria from a specialized cancer hospital in 2013 in order to provide a basis for rational clinical antimicrobial agents. Methods： Pathogenic bacteria identification and drug sensitivity tests were performed with a VITEK 2 compact automatic identification system and data were analyzed using WHONET5.6 software.Results： Of the 1,378 strains tested, 980 were Gram-negative bacilli, accounting for 71.1%, in which Klebsiella pneumonia, Escherichia coli and Pseudomonas aeruginosa were the dominant strains. We found 328 Gram-positive coccus, accounting for 23.8%, in which the amount of Staphylococcus aureus was the highest. We identified 46 fungi, accounting for 4.1%. According to the departmental distribution within the hospital, the surgical departments isolated the major strains, accounting for 49.7%. According to disease types, lung cancer, intestinal cancer and esophagus cancer were the top three, accounting for 20.9%, 17.3% and 14.2%, respectively. No strains were resistant to imipenem, ertapenem or vancomycin.Conclusions： Pathogenic bacteria isolated from the specialized cancer hospital have different resistance rates compared to commonly used antimicrobial agents; therefore antimicrobial agents to reduce the morbidity and mortality of infections should be used.

Fluorescent antibiotics for real-time tracking of pathogenic bacteria

The harm of pathogenic bacteria to humans has promoted extensive research on physiological processes of pathogens,such as the mechanism of bacterial infection,antibiotic mode of action,and bacterial antimicrobial resistance.Most of these processes can be better investigated by timely tracking of fluorophore-derived antibiotics in living cells.In this paper,we will review the recent development of fluorescent antibiotics featuring the conjugation with various fluorophores,and focus on their applications in fluorescent imaging and real-time detection for various physiological processes of bacteria in vivo.

Functional identification of phenazine biosynthesis genes in plant pathogenic bacteria Pseudomonas syringae pv. tomato and Xanthomonas oryzae pv. oryzae

Phenazines are secondary metabolites with broad spectrum antibiotic activity and thus show high potential in biological control of pathogens. In this study, we identified phenazine biosynthesis （phz） genes in two genome-completed plant pathogenic bacteria Pseudomonas syringae pv. tomato （Pst） DC3000 and Xanthomonas oryzae pv. oryzae （Xoo） PXO99A. Unlike the phz genes in typical phenazine-producing pseudomonads, phz homologs in Pst DC3000 and Xoo PXO99A consisted of phzC/D/E/F/G and phzC/E1/E2/F/G, respectively, and the both were not organized into an operon. Detection experiments demonstrated that phenazine-l-carboxylic acid （PCA） of Pst DC3000 accumulated to 13.4 IJg L-1, while that of Xoo PXO99A was almost undetectable. Moreover, Pst DC3000 was resistant to 1 mg mL-1 PCA, while Xoo PXO99A was sensitive to 50 IJg mL ~ PCA. Furthermore, mutation of phzF blocked the PCA production and significantly reduced the pathogenicity of Pst DC3000 in tomato, while the complementary strains restored these phenotypes. These results revealed that Pst DC3000 produces low level of and is resistant to phenazines and thus is unable to be biologically controlled by phenazines. Additionally, phz-mediated PCA production is required for full pathogenicity of Pst DC3000. To our knowledge, this is the first report of PCA production and its function in pathogenicity of a plant pathogenic P. syringae strain.

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.

Antibacterial and Antifungal Activity of Organic and Peptidic Extracts of Ecuadorian Endophytic Fungi

Antibacterial and antifungal activities of 32 organic and 14 peptidic extracts obtained from twelve endophytic fungi of the Collection of Endophytes Quito-Catolica were tested against the pathogenic bacteria Staphylococcus aureus, Pseudomonas aeruginosa, Serratia marcescens, Escherichia coli and Salmonella enteritidis, and against the fungal oomycete Pythium ultimum. It was observed that the extracts of Marasmiellus candidus (CEQCA-O1113), Xylaria laevis (CEQCA-O1399), Fusarium solani (CEQCA-O1393), Diaporthe helianthi (CEQCA-O1394) and Xylaria sp. (CEQCA-O1400) partially or totally inhibited the microorganisms tested. The extracts CEQCA-O1399.P1 and CEQCA-O1113.D1 showed fungistatic and fungicidal effects against P. ultimum, respectively. Six extracts strongly inhibited S. aureus, and E. coli was strongly inhibited by the extract CEQCA-O1113.E2. Based on a bioassay-guided approach, the extracts with strong bioactivity in the antibacterial assays by an agar diffusion method were also tested by a disk diffusion method, and the minimal inhibitory concentrations were determined by a microdilution assay. The most bioactive extract, CEQCA-O1113.D1, was also purified by high performance liquid chromatography. The extract recovered its initial bioactivity against S. aureus in two fractions after the purification. Overall, the results of this study highlight the potential of the fungal endophytes as producers of bioactive compounds.

Synergistic Effect of Combined Antibiotics against Some Selected Multidrug Resistant Human Pathogenic Bacteria Isolated from Poultry Droppings in Akure, Nigeria

Antibiotic resistant bacteria pass between humans, between animals and between humans and animals in both directions, the use of antibiotics in poultry has contributed to multiple antibiotic resistant in pathogenic bacteria and use of two antibiotics might prevent the emergence of resistance to either. In this study, synergistic effect of combined antibiotics against multidrug resistant human pathogenic bacterial isolates from poultry droppings in Akure, Nigeria was examined. Collection of samples, isolation and identification of bacteria were carried out using standard microbiological method, antibiotic sensitivity test was performed by disc diffusion method and zone of inhibition was used to interpret the sensitivity test as resistant, susceptible or intermediate while combined effects of two antibiotics were investigated by macrobroth dilution and checkerboard assay methods while the synergetic effects of combined antibiotics were calculated using Fractional Inhibitory Concentration (FIC) and percentage synergistic interaction was calculated. All the ten (10) species of bacterial isolates were multidrug resistant and are less resistant to ofloxacin. The highest percentage synergistic interactions observed were Ofloxacin + Amoxicillin (90%), Ciprofloxacin + Amoxicillin (90%), Tetracycline + Amoxicillin (70%), Tetracycline + Augmentin (80%), Cotrimoxazol + Amoxicillin (50%), Cotrimoxazol + Augmentin (70%), Chloramphenicol + Amoxicillin (70%) and Chloramphenicol + Augmentin (80%). Poultry droppings is a potential source of human pathogenic bacteria, high frequency of multiple antibiotic resistance bacteria observed in this study is of great treat to man as this may cause the treatment of infection caused by these bacteria to be difficult. Combination of beta-lactam antibiotic with fluoroqunolones, tetracycline, Chloramphenicol and Cotrimoxazole was synergetic and this will reduce dose related toxicity and prevent resistance to single antibiotic.

Rapid label-free SERS detection of foodborne pathogenic bacteria based on hafnium ditelluride-Au nanocomposites

Two-dimensional(2D)nanomaterials have captured an increasing attention in biophotonics owing to their excellent optical features.Herein,2D hafnium ditelluride(HfTe_(2)),a new member of transition metal tellurides,is exploited to support gold nanoparticles fabricating HfTe_(2)-Au nanocomposites.The nanohybrids can serve as novel 2D surface-enhanced Raman scattering(SERS)substrate for the label-free detection of analyte with high sensitivity and reproducibility.Chemical mechanism originated from HfTe_(2) nanosheets and the electromagnetic enhancement induced by the hot spots on the nano-hybrids may largely contribute to the superior SERS effect of HfTe_(2)-Au nanocomposites.Finally,HfTe_(2)-Au nanocomposites are utilized for the label-free SERS analysis of foodborne pathogenic bac-teria,which realize the rapid and ultrasensitive Raman test of Escherichia coli,Listeria mono-cytogenes,Staphylococcus aureus and Salmonella with the limit of detection of 10 CFU/mL and the maximum Raman enhancement factor up to 1.7×10^(8).Combined with principal component analysis,HfTe_(2)-Au-based SERS analysis also completes the bacterial classification without extra treatment.

Antibacterial activity of plant extracts in different solvents against pathogenic bacteria: An in vitro experiment

Objective:To assess the antibacterial activity of 5 selected plants against 4 pathogenic bacteria.Methods:Three solvents with different polarities were used to extract antimicrobial agents from the plants via maceration technique.The agar-disc diffusion technique was adopted to primarily screen antibacterial activities.Broth-dilution assay was employed to determine the minimum inhibitory concentration(MIC)and the minimum bactericidal concentration(MBC).Results:Among all extracts,the ethanol extract of Piper betle Linn showed the highest antibacterial activity against Gram-positive and the negative bacteria.MIC and MBC of the ethanol extract of Piper betle Linn against Salmonella typhimurium were the same(1?562.50 mg/L);while it showed the highest MIC and MBC against Pseudomonas aeruginosa of 6?250 mg/L and 12?500 mg/L,respectively.Conclusions:Salmonella typhimurium is the most susceptible bacteria while Pseudomonas aeruginosa is the most resistant bacteria towards the ethanol extract of Piper betle Linn.Piper betle possesses compounds with potential antibacterial activity and might be useful as an alternative to control infectious diseases.

Effect Evaluation about Pathogenic Bacteria of Diary Cow Mastitis Treated with Ruyuankang

In this research,36 cows with clinical mastitis were treated with the Chinese herb injection Ruyuankang which was developed by our group.Milk samples of pre and post treatment of Ruyuankang were collected for isolation and characterization of pathogenic bacteria.The dynamic changes of pathogenic bacteria were investigated as well.The results indicated that Ruyuankang could effectively inhibit the common pathogenic bacteria of mastitis which result in the negative rate of pathogenic bacteria 71.74%.

Effect of Chinese Herbal Medicine on Pathogenic Bacteria of Cow Mastitis

[Objective]The paper was to analyze the clinical therapeutic effects of Chinese herbal medicine on pathogenic bacteria of cow mastitis.[Method]Eight Chinese herbs and three compound extracts were selected for the treatment of cow mastitis.Four kinds of pathogenic bacteria were successfully isolated from cows with mastitis by flat perforation irrigation method and tube double dilution method.In addition,in vitro antibacterial test was carried out,and in vitro inhibitory effects of single and compound Chinese medicine extracts on pathogenic bacteria causing cow mastitis were studied.[Result]A total of seven Chinese herbs had good inhibitory effect,such as Taraxacum mongolicum,Viola yedonensis and Polygala japonica,which received good effect of activating blood circulation and removing blood stasis,clearing away heat and toxic materials.[Conclusion]As good Chinese herbal medicines,T.mongolicum,V.yedonensis and P.japonica could be popularized.