The virulence regulator AbsR in avian pathogenic Escherichia coli has pleiotropic effects on bacterial physiology

Avian pathogenic Escherichia coli(APEC)belonging to extraintestinal pathogenic E.coli(ExPEC)can cause severe infections in extraintestinal tissues in birds and humans,such as the lungs and blood.MprA(microcin production regulation,locus A,herein renamed AbsR,a blood survival regulator),a member of the MarR(multiple antibiotic resistance regulator)transcriptional regulator family,governs the expression of capsule biosynthetic genes in human ExPEC and represents a promising druggable target for antimicrobials.However,a deep understanding of the AbsR regulatory mechanism as well as its regulon is lacking.In this study,we present a systems-level analysis of the APEC AbsR regulon using ChIP-Seq(chromatin immunoprecipitation sequencing)and RNA-Seq(RNA sequencing)methods.We found that AbsR directly regulates 99 genes and indirectly regulates 667 genes.Furthermore,we showed that:1)AbsR contributes to antiphagocytotic effects by macrophages and virulence in a mouse model for systemic infection by directly activating the capsular gene cluster;2)AbsR positively impacts biofilm formation via direct regulation of the T2SS(type II secretion system)but plays a marginal role in virulence;and 3)AbsR directly upregulates the acid tolerance signaling system EvgAS to withstand acid stress but is dispensable in ExPEC virulence.Finally,our data indicate that the role of AbsR in virulence gene regulation is relatively conserved in ExPEC strains.Altogether,this study provides a comprehensive analysis of the AbsR regulon and regulatory mechanism,and our data suggest that AbsR likely influences virulence primarily through the control of capsule production.Interestingly,we found that AbsR severely represses the expression of the type I-F CRISPR(clustered regularly interspaced short palindromic repeats)-Cas(CRISPR associated)systems,which could have implications in CRISPR biology and application.

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.

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.

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.

Antibacterial activity of lemongrass(Cymbopogon citratus) oil against some selected pathogenic bacterias

Objective:To find the effectiveness of essential oil of lemongrass for the treatment of pathogenic organisms.Methods:Lemongrass oil was investigated for activity against Staphylococcus aureus (S.aureus),Bacillus cereus(B.cereus),Bacillus subtilis(B.subtilis),Escherichia coli(E.coli), Klebsiella pneumoniae(K.pneumoniae) and Pseudomonas aeruginosa(P.aeruginosa),using Agar Diffusion Method and Broth Dilution Method.Minimum inhibitory concentration(MIC) and minimum bactericidal concentration(MBC) were determined by the Broth Dilution Method. The antibiotic susceptibility test against the test organisms was performed by Disc Diffusion Method.Results:Lemongrass was found effective against all the test organisms except P. aeruginosa.Gram positive organisms were found more sensitive to lemon grass oil as compared to gram negative organisms.The test organisms were found inhibited by Lemon grass oil at lower concentrations in Broth Dilution Method as compared to Agar Diffusion Method.Conclusions: The tested organisms,particularly gram-negative organisms had shown high resistance towards different antibiotics whereas they were found to be inhibited by lemongrass oil even at lower concentration.Thus lemongrass oil is effective against drug resistant organisms.It can be suggested that use of lemongrass oil would be helpful in the treatment of infections caused by multidrug resistant organisms.

Improving Geese Production Performance in “Goose-Fish” Production System by Competitive Reduction of Pathogenic Bacteria in Pond Water

This study investigated whether competitive reduction of pathogenic bacteria growth in pond water alleviates lipopolysaccharide （LPS） contamination and improves geese production performances in the ＂goose-fish＂ production system, thereby providing the potential for an improved technique for ecological water fowl production. In the first experiment, 240 Magang goslings of 15-d age were randomly and equally allocated into 16 ＂yard and pond＂ pens using a 2-2 factorial design with 4 replications per treatment. In the 55-d experimental period, the goslings received 2 main treatments： supplementation of Bacillus subtilis spores in the feed and addition of photosynthetic bacteria （PSB） to the pond water. Both B. subtilis spores and PSB treatments significantly suppressed water counts of Gram-negative bacteria Escherichia coli, Salmonella and Shigella, and LPS concentrations in pond water and in gosling blood （P〈0.05）. As the result, the two treatments significantly improved gosling weight gain and carcass quality, marked by enhanced breast and leg muscle percentages and reduced subcutaneous fat proportions （P〈0.05）. Moreover, the improved effects of B. subtilis spores and PSB treatments were additive. In the second experiment, 1 160 adult geese were induced to start egg laying from May throughout the summer months. The geese were separated equally into control and experimental flocks to fit into 2 integration production units, with a density of 1 bird m-2 meter on pond water. Experimental flock geese were treated with B. subtilis spores in feed and PSB in the pond water for the duration of the study. Such treatment combination significantly depressed the growth of E. coli, Salmonella and Shigella in the pond water and reduced LPS concentrations both in pond water and in geese blood （P〈0.01）. As a result, egg fertility, fertile and set egg hatchabilities were all improved in the treated flock. Results from both growing goslings and breeding geese demonstrated that water bacteria pollution can be competitively reduced by supplementation with B. subtilis spores via the feed and addition of PSB in pond water, each of which reduces LPS contamination to geese and improves production performances. Micro- ecological agents such as B. subtilis spores and PSB improve water quality and provide a simple ecological technique for the ＂water fowl-fish＂ integrative production system.

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.

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.

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.

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.

Synergistic effect between cryptotanshinone and antibiotics in oral pathogenic bacteria

Cryptotanshinone (CT), a major tanshinone of medicinal plant Salvia miltiorrhiza Bunge, demonstrated effective in vitro antibacterial activity against all oral bacteria tested in this experiment. The antibacterial activities of CT against oral bacteria were assessed using the checkerboard and time-kill methods to evaluate the synergistic effects of treatment with ampicillin or gentamicin. The CT was determined against oral pathogenic bacteria with MIC and MBC values ranging from 0.5 to 16 and 1 to 64 μg/mL;for am- picillin from 0.0313 to 16 and 0.125 to 32 μg/mL;for gentamicin from 2 to 256 and 4 to 512 μg/mL respectively. The range of MIC50 and MIC90 were 0.0625 - 8 μg/mL and 1 - 64 μg/mL, respectively. The combination effects of CT with antibiotics were synergistic (FIC index < 0.5) against tested oral bacteria except additive, Streptococcus sobrinus, S. criceti, and Actinobacillus actinomycetemcomitans (FIC index < 0.75 - 1.0). The MBCs were shown reducing ≥4 - 8-fold, indicating a synergistic effect as defined by a FBCI of ≤0.5. Furthermore, a time-kill study showed that the growth of the tested bacteria was completely attenuated after 3 - 6 h of treatment with the 1/2 MIC of CT, regardless of whether it was administered alone or with ampicillin or gentamicin. The results suggest that CT could be employed as a natural antibacterial agent against cariogenic and periodontopathogenic bac- teria.

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.

Species Identification of Mealie Meal Spoilage Organisms and Pathogenic Bacteria from Selected Food Stores in Lusaka District of Zambia

Background: Mealie Meal is one of the by-products of maize from dry milling. It constitutes more significant part of the daily diet of growing population of the Lusaka District of Zambia. However, reported cases of a cholera outbreak in October 2017-May 2018 were attributed to water contamination in the households of Lusaka. Hitherto there is no previous study or documented data for gastroenteritis caused by microbial contamination from food stores. Aim: The main objective of the study was to carry out species identification of Mealie meal spoilage organisms and pathogenic bacteria from selected food stores in Lusaka district from September 2019-March 2020. Crosssectional survey was conducted for food spoilage organisms and pathogenic bacteria of packaged maize meal flour from a selected food store. The number of packaged Mealie meal samples from selected food stores included in the study was 143 samples. Samples were obtained by a simple stratified random selection from food stores in Lusaka District. Thus Mealie Meal samples bought from Street food vendors were 96 (67%), Shops—30 (21%), Malls—17 (11.9%). Subsequently isolation by spread and pour plate methods, and species identification of microbial contamination by Cell Culture processes coupled to microbial morphological and biochemical characterization was performed. Microsoft Office Excel (version 2007) Spreadsheet was employed for the categorical data analysis and summarized graphical presentation of data. Laboratory findings revealed: Spoilage organisms—Aspergillum species, Mucor Species, Candida Species, Clostridium Species and Bacillus Species;Pathogenic Bacteria Species—Clostridium perfringens, Staphylococcus aureus, Clostridium tetani, Bacillus, Escherichia coli, Pseudomonas aeruginosa and Yesinia pestis. It can be established from the laboratory results earlier mentioned that the food poisoning outbreak could evolve from food stores in Lusaka district apart from water contamination.

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.

Application of 16s rDNA Sequencing in the Analysis of Pathogenic Bacteria in Sputum of Severe Bacterial Pneumonia

Objective: 120 patients with severe pneumonia who were kept in the comprehensive ICU of our hospital in 2018 were selected, and 16s rDNA sequencing was performed to analyze the composition of pathogenic bacteria in the sputum of severe pneumonia. Methods: The sputum samples of patients with severe bacterial pneumonia were collected, and the diversity of pathogens in the samples was analyzed by polymerase chain reaction (PCR) amplification and high-throughput sequencing (16s rDNA PCR-DGGE). Results: Sequence showed that sputum samples contained a relatively large number of species, and there were many species that were not detected by sequencing. The dominant bacteria were Streptococcus, Sphingomonas, Corynebacterium, Denatobacteria, Aquobacteria, Acinetobacteria, Prevotella, Klebsiella, Pseudomonas, etc. Conclusion: Bacteria caused by sputum of severe bacterial pneumonia are complex and diverse, which provides new methods and ideas for individualized treatment of patients with severe pneumonia.

Application of 16S rDNA Sequencing Technology in the Analysis of Pathogenic Bacteria in Sputum of Severe Pneumonia

The diagnosis of pathogenic bacteria in severe pneumonia is difficult and the prognosis is poor. Its outcome is closely related to bacterial pathogenicity and the timeliness and pertinence of antibiotic treatment. Therefore, early diagnosis is of great significance to the prognosis of patients. Sputum examination and culture is the gold standard for the diagnosis of pathogens of severe pneumonia. However, due to the long time of bacterial culture, the early use of antibiotics, the change of bacteria species, mixed infection and other problems, the results of bacterial culture in sputum are often false negative. With the continuous application of new molecular biology techniques in clinical detection, the classification of bacteria and microorganisms has deepened from the identification of phenotypic characteristics to the classification of gene characteristics. Sequencing analysis with 16S rDNA sequencing technology has the characteristics of high sequencing flux, large amount of data obtained, short cycle, and can more comprehensively reflect the species composition of microbial community, real species distribution and abundance information. In this paper, 16S rDNA sequencing technology was used to analyze the bacterial population composition in the sputum of severe pneumonia, and to explore a new method of etiological diagnosis.