Sequencing and multiple antimicrobial resistance of Pseudomonas fluorescens isolated from Nile tilapia fish in Egypt

Abstract Pseudomonas fluorescens is one of the main causes of septicemic diseases among freshwater fish, causing severe economic losses and decreasing farm efficiency. Thus, this research was aimed to investigate the occurrence of P. fluorescens in Nile Tilapia (O. niloticus) fish in Egypt, gene sequencing of 16SrDNA gene, and antimicrobial susceptibility. P. fluorescens strains were detected in 32% (128/400) of apparently healthy (9%; 36/400) and diseased (23%; 92/400) Nile tilapia fish. The highest prevalence was observed in gills of fish, 31.3% followed by intestine 26.9%, liver 24.2%, and kidneys 17.6%. The PCR results for the 16SrDNA gene of P. fluorescens showed 16SrDNA gene in 30% of examined isolates. Moreover, Homogeny and a strong relationship between strains of P. fluorescens was confirmed using 16SrDNA sequences. Beside the responsibility of 16SrDNA gene on the virulence of P. fluorescens. The results of antimicrobial susceptibility tests revealed that all strains were resistant to piperacillin (100%), followed by ceftazidime (29.7%), and cefepime (25.8%). The strains of P. fluorescence were highly sensitive to cefotaxime (74.2%), followed by ceftriaxone and levofloxacin (70.3% each). Interestingly, 29.7% of strains of P. fluorescens were multiple antimicrobial-resistant (MAR).

Resumo Pseudomonas fluorescens é uma das principais causas de doenças septicêmicas em peixes de água doce, causando graves perdas econômicas e diminuindo a eficiência da fazenda. Assim, esta pesquisa teve como objetivo investigar a ocorrência de P. fluorescens em peixes de tilápia-do-nilo (O. niloticus) no Egito, sequenciamento do gene 16S rDNA e suscetibilidade antimicrobiana. Cepas de P. fluorescens foram detectadas em 32% (128/400) de peixes tilápia-do-nilo aparentemente saudáveis (9%; 36/400) e doentes (23%; 92/400). A maior prevalência foi observada nas brânquias dos peixes, 31,3%, seguida pelo intestino 26,9%, fígado 24,2% e rins 17,6%. Os resultados da PCR para o gene 16SrDNA de P. fluorescens mostraram o gene 16SrDNA em 30% dos isolados examinados. Além disso, a homogeneidade e uma forte relação entre cepas de P. fluorescens foi confirmada usando sequências de 16SrDNA. Além da responsabilidade do gene 16SrDNA na virulência de P. fluorescens. Os resultados dos testes de suscetibilidade antimicrobiana revelaram que todas as cepas foram resistentes à piperacilina (100%), seguida pela ceftazidima (29,7%) e cefepima (25,8%). As cepas de P. fluorescens foram altamente sensíveis à cefotaxima (74,2%), seguida pela ceftriaxona e levofloxacina (70,3% cada). Curiosamente, 29,7% das cepas de P. fluorescens eram multirresistentes a antimicrobianos (MAR).

Sequencing and multiple antimicrobial resistance of Pseudomonas fluorescens isolated from Nile tilapia fish in Egypt / Sequenciamento e resistência antimicrobiana múltipla de Pseudomonas fluorescens isoladas de tilápia-do-nilo no Egito

Pseudomonas fluorescens is one of the main causes of septicemic diseases among freshwater fish, causing severe economic losses and decreasing farm efficiency. Thus, this research was aimed to investigate the occurrence of P. fluorescens in Nile Tilapia (O. niloticus) fish in Egypt, gene sequencing of 16SrDNA gene, and antimicrobial susceptibility. P. fluorescens strains were detected in 32% (128/400) of apparently healthy (9%; 36/400) and diseased (23%; 92/400) Nile tilapia fish. The highest prevalence was observed in gills of fish, 31.3% followed by intestine 26.9%, liver 24.2%, and kidneys 17.6%. The PCR results for the 16SrDNA gene of P. fluorescens showed 16SrDNA gene in 30% of examined isolates. Moreover, Homogeny and a strong relationship between strains of P. fluorescens was confirmed using 16SrDNA sequences. Beside the responsibility of 16SrDNA gene on the virulence of P. fluorescens. The results of antimicrobial susceptibility tests revealed that all strains were resistant to piperacillin (100%), followed by ceftazidime (29.7%), and cefepime (25.8%). The strains of P. fluorescence were highly sensitive to cefotaxime (74.2%), followed by ceftriaxone and levofloxacin (70.3% each). Interestingly, 29.7% of strains of P. fluorescens were multiple antimicrobial-resistant (MAR).

Pseudomonas fluorescens é uma das principais causas de doenças septicêmicas em peixes de água doce, causando graves perdas econômicas e diminuindo a eficiência da fazenda. Assim, esta pesquisa teve como objetivo investigar a ocorrência de P. fluorescens em peixes de tilápia-do-nilo (O. niloticus) no Egito, sequenciamento do gene 16S rDNA e suscetibilidade antimicrobiana. Cepas de P. fluorescens foram detectadas em 32% (128/400) de peixes tilápia-do-nilo aparentemente saudáveis ​​(9%; 36/400) e doentes (23%; 92/400). A maior prevalência foi observada nas brânquias dos peixes, 31,3%, seguida pelo intestino 26,9%, fígado 24,2% e rins 17,6%. Os resultados da PCR para o gene 16SrDNA de P. fluorescens mostraram o gene 16SrDNA em 30% dos isolados examinados. Além disso, a homogeneidade e uma forte relação entre cepas de P. fluorescens foi confirmada usando sequências de 16SrDNA. Além da responsabilidade do gene 16SrDNA na virulência de P. fluorescens. Os resultados dos testes de suscetibilidade antimicrobiana revelaram que todas as cepas foram resistentes à piperacilina (100%), seguida pela ceftazidima (29,7%) e cefepima (25,8%). As cepas de P. fluorescens foram altamente sensíveis à cefotaxima (74,2%), seguida pela ceftriaxona e levofloxacina (70,3% cada). Curiosamente, 29,7% das cepas de P. fluorescens eram multirresistentes a antimicrobianos (MAR).

Efficacy of Chinese Medicinal Formulae for Antimicrobial-resistant Pneumonia: A Systematic Review and Meta-analysis / 中国实验方剂学杂志

Objective:To evaluate the efficacy and safety of Chinese medicinal formulae in the treatment of antimicrobial-resistant pneumonia. Method:Following article retrieval from eight databases and data extraction by two reviewers， the methodological quality of the included trials was assessed and the outcome indicators were subjected to Meta-analysis using RevMan 5.3. Result:A total of 24 randomized controlled trials （RCTs） were included， involving 1 818 cases. Meta-analysis showed that Chinese medicinal formulae combined with western routine intervention was superior to the western routine intervention in improving the overall response rate （ORR） ［relative risk （RR）=1.27， 95% confidence interval （CI） （1.21， 1.34）， <inline-formula><alternatives><mml:math xmlns:mml="http://www.w3.org/1998/Math/MathML" id="M5"><mml:mi>P</mml:mi></mml:math><graphic specific-use="big" xmlns:xlink="http://www.w3.org/1999/xlink" xlink:href="alternativeImage/27038DAF-2FF7-4d58-8001-0E6465A33408-M005.jpg"><?fx-imagestate width="2.28600001" height="2.62466669"?></graphic><graphic specific-use="small" xmlns:xlink="http://www.w3.org/1999/xlink" xlink:href="alternativeImage/27038DAF-2FF7-4d58-8001-0E6465A33408-M005c.jpg"><?fx-imagestate width="2.28600001" height="2.62466669"?></graphic></alternatives></inline-formula><0.000 01］， the bacterial clearance rate ［RR=1.49，95% CI （1.33， 1.66）， <inline-formula><alternatives><mml:math xmlns:mml="http://www.w3.org/1998/Math/MathML" id="M6"><mml:mi>P</mml:mi></mml:math><graphic specific-use="big" xmlns:xlink="http://www.w3.org/1999/xlink" xlink:href="alternativeImage/27038DAF-2FF7-4d58-8001-0E6465A33408-M006.jpg"><?fx-imagestate width="2.28600001" height="2.62466669"?></graphic><graphic specific-use="small" xmlns:xlink="http://www.w3.org/1999/xlink" xlink:href="alternativeImage/27038DAF-2FF7-4d58-8001-0E6465A33408-M006c.jpg"><?fx-imagestate width="2.28600001" height="2.62466669"?></graphic></alternatives></inline-formula><0.000 01］， and the clinical pulmonary infection score （CPIS） ［mean difference （MD）=-1.64， 95% CI （-1.87， -1.41）， <inline-formula><alternatives><mml:math xmlns:mml="http://www.w3.org/1998/Math/MathML" id="M7"><mml:mi>P</mml:mi></mml:math><graphic specific-use="big" xmlns:xlink="http://www.w3.org/1999/xlink" xlink:href="alternativeImage/27038DAF-2FF7-4d58-8001-0E6465A33408-M007.jpg"><?fx-imagestate width="2.28600001" height="2.62466669"?></graphic><graphic specific-use="small" xmlns:xlink="http://www.w3.org/1999/xlink" xlink:href="alternativeImage/27038DAF-2FF7-4d58-8001-0E6465A33408-M007c.jpg"><?fx-imagestate width="2.28600001" height="2.62466669"?></graphic></alternatives></inline-formula><0.000 01］. There was no significant difference in the incidence of adverse reactions ［RR=0.72， 95% CI （0.48， 1.07），<inline-formula><alternatives><mml:math xmlns:mml="http://www.w3.org/1998/Math/MathML" id="M8"><mml:mtext> </mml:mtext><mml:mi>P</mml:mi></mml:math><graphic specific-use="big" xmlns:xlink="http://www.w3.org/1999/xlink" xlink:href="alternativeImage/27038DAF-2FF7-4d58-8001-0E6465A33408-M008.jpg"><?fx-imagestate width="3.04799986" height="2.62466669"?></graphic><graphic specific-use="small" xmlns:xlink="http://www.w3.org/1999/xlink" xlink:href="alternativeImage/27038DAF-2FF7-4d58-8001-0E6465A33408-M008c.jpg"><?fx-imagestate width="3.04799986" height="2.62466669"?></graphic></alternatives></inline-formula>=0.1］. The comparison with the western routine intervention also revealed that Chinese medicinal formulae better improved the ORR and CPIS. Conclusion:According to the current research results， the Chinese medicinal formulae alone or combined with western routine intervention yielded more favorable clinical outcomes than western routine intervention in the treatment of antimicrobial-resistant pneumonia， without increasing the incidence of adverse events. Due to limited quality and quantity of the included RCTs， more high-quality trials are required to verify the above conclusions.

Isolation, identification and antibiotic sensitivity pattern of aerobic bacteria from burn wound patient admitted in tertiary care hospital

Background: Significant burn injuries induce a state of immunosuppression that predisposes patients to infectious complications, thus the rate of nosocomial infections are higher. Rapidly merging multidrug resistant among the various isolate in indoor burn patients are depending on time-line becoming serious threat for managing therapeutically. Objective of this study is to determine the aetiological factor, prevalence, antimicrobial susceptibility pattern and emerging nosocomial pathogens.Methods: A prospective study was carried in burn ward of K.L.E.’s Dr. Prabhakar Kore Hospital and Medical Research Centre, Belgaum for the period of 1 year. Pair of wound swab were collected from patient having burnt more than 30% (RULE OF NINE) on 3rd day of stay. Sample were collected aseptically from 30 patients and processed by convectional culture and biochemical identification procedures and tested against commonly used antibiotics.Results: 30 patients that fall under inclusive criteria were enrolled in the study. The total burn surface area (TBSA) ranges from 30-82%. The ratio of female to male patient suffering burn wound in our study is 1.5:1. Aetiology of burn is heat (moist/dry) mostly. Depending upon degree of burn, most of patient suffered from 20 degree (superficial to deep) injury. From 30 swab cultures, 42 isolates were identified during the study in which mixed were 66.66% and one is fungi. The most commonly isolated is Pseudomonas aeruginosa (45.24%) then Klebsiella pneumoniae (19.04%), Acinetobacter spp. (14.28%), Staphylococccus aureus (11.90%). Among gram positive isolates, isolates are found to be most resistant to Erythromycin (100%) and Co-trimoxazole (100%) and sensitive to Vancomycin (71.42%). Among gram negative isolates are found to be most resistant to Gentamicin (91.65%), Ciprofloxacin (82.35%), Ceftazidime (82.35%) and sensitive to Meropenem (52.95%), Piperacillin (35.30%), Carbenicillin (29.41%).Conclusions: Pseudomonas aeruginosa was found to be the most common isolate. The nature of microbial wound colonization and flora changes with time should be taken into consideration in empirical antimicrobial therapy.

Genome sequence of Prevotella intermedia SUNY aB G8-9K-3, a biofilm forming strain with drug-resistance

Abstract Prevotella intermedia has long been known to be as the principal etiologic agent of periodontal diseases and associated with various systemic diseases. Previous studies showed that the intra-species difference exists in capacity of biofilm formation, antibiotic resistance, and serological reaction among P. intermedia strains. Here we report the genome sequence of P. intermedia SUNY aB G8-9K-3 (designated ATCC49046) that displays a relatively high antimicrobial resistant and biofilm-forming capacity. Genome sequencing information provides important clues in understanding the genetic bases of phenotypic differences among P. intermedia strains.

COMMON INFECTIONS IN THE ELDERLY IN THE HOME OR NURSING HOME SETTING

Infections in the elderly are associated with high morbidity and mortality. Diagnosing infections in the elderly is challenging due to their atypical and subtle presentation. A high index of suspicion is often needed. Commonly encountered infections in the elderly include bacterial pneumonia, urinary tract infection, cellulitis and Herpes zoster. In addition, institutionalised elderly and those with multiple hospital admissions are at risk of infection with Multidrug-resistant Organisms (MROs); this can be difficult to manage. The purpose of this article is to look at some common infections in the elderly encountered in the home or nursing home, and review their management.

Analysis of the Antimicrobial-Resistant mechanism and clinical test of Staphylococcus Aureus / 国际检验医学杂志

Objective To investigate the antimicrobial‐resistant profile and genes carried by 80 staphylococcus aureus and analy‐size its antimicrobial‐resistant mechanism .Methods The bacteria identification and the antimicrobial susceptibility test were con‐ducted by VITEK‐2 compact automatic system .Methicillin resistant taphylococcus aureus (MRSA) were screened by disk diffusion method with cefoxitin .The polymerase chain reaction(PCR)was used to detect the antimicrobial‐resistant genes .Results The re‐sistance rates of 80 staphylococcus aureus to penicillin ,erythromycin and gentamicin were 91 .25% ,85 .0% and 71 .25% ,respective‐ly .All of the isolates were susceptible to linezolid ,vancomycin ,teicoplanin and tigecycline .Among the 80 isolates ,there were 22 MRSA ,accounting for 27 .5% ,and 13 were inducible antimicrobial‐resistant .57 (71 .25% ) carried antimicrobial‐resistant genes ,in‐cluding 6 genes .Of which ,21(26 .25% )carried ermB and aac(6′)/aph(2) simultaneously ,9(11 .25% )carried ermA ,mecA ,qacA simultaneously ,and all of the 9 were MRSA ,13(16 .25% ) only carried ermC .Conclusion The resistance rates of staphylococcus aureus were high to penicillin ,erythromycin and gentamicin .The various antimicrobial‐resistant genes were positive in staphylococ‐cus aureus .We should pay attention to the detection of the antimicrobial‐resistant gene and choose antibacterial drug rationally .

Antimicrobial Producing Bacteria Isolated from Tropical Peat Swamp Soil

Aims: The aims of this study were to isolate and characterize antimicrobial producing bacteria from tropical peat swamp forest soils. Methodology and results: Bacteria isolated from peat soil were screened for antimicrobial properties via agar overlay assay. Broth microdilution was performed using crude-cell free supernatant (CCFS) to determine the minimum inhibitory concentration (MIC). One isolate was selected due to its broad spectrum activity and identified as Burkholderia spp. with a maximum identity of 99% via 16s rRNA gene PCR. This isolate was able to produce antimicrobials that were active against several Gram positive bacteria, Gram negative bacteria and also yeast. The antimicrobial activity of the CCFS was stable at a pH range of 1 to 11, temperatures of −20 °C to 80 °C, and after treatment with several proteolytic enzymes: α-chymotrypsin, proteinase K and trypsin, indicating that the antimicrobial produced might not be proteinaceous in nature. It is possible that the isolate can produce polyketides, a type of antimicrobial compound produced by Burkholderia known to be resistant to proteolytic enzymes. However, further work needs to be done to confirm this. Conclusion, significance and impact of study: The presence of antimicrobial producing bacteria signified that tropical peat swamps are indeed a potential source for antimicrobials to combat infections.

Antimicrobial susceptibility and pathogenic genes of Staphylococcus aureus isolated from the oral cavity of patients with periodontitis

PURPOSE: The goal of this study was to characterize the patterns of antimicrobial resistance and virulence genes in samples of Staphylococcus aureus (S. aureus) isolated from periodontitis patients. METHODS: From July 2015 to August 2015, oral saliva was collected from a total of 112 patients diagnosed with periodontitis, including 80 outpatients in dental hospitals and 32 patients in dental clinics located in Seoul and Cheonan. The samples were subjected to a susceptibility test to evaluate the prevalence of antimicrobial resistance, and the pathogenic factors and antimicrobial resistance factors in the DNA of S. aureus were analyzed using polymerase chain reaction. RESULTS: A susceptibility test against 15 antimicrobial agents showed that 88% of cultures were resistant to ampicillin, 88% to penicillin, and 2% to oxacillin. Resistance to at least two drugs was observed in 90% of cultures, and the most common pattern of multidrug resistance was to ampicillin and penicillin. Enterotoxins were detected in 65.9% of samples. The cell hemolysin gene hld was detected in 100% of cultures and hla was detected in 97.6% of samples. All strains resistant to penicillin and ampicillin had the blaZ gene. The aph(3')IIIa gene, which encodes an aminoglycoside modifying enzyme, was detected in 46.3% of samples. CONCLUSIONS: In the treatment of oral S. aureus infections, it is important to identify the pathogenic genes and the extent of antimicrobial resistance. Furthermore, it is necessary to study patterns of antimicrobial resistance and cross-infection in the context of periodontological specialties in which antimicrobials are frequently used, such as maxillofacial surgery, where the frequency of antimicrobial use for minor procedures such as implant placement is increasing.

Resistant Mechanism and Strain Relationship of Multi-resistant Pseudomonas aeruginosa / 中华医院感染学杂志

OBJECTIVE To investigate the correlated resistant genes encoding ?-lactamases,aminoglycoside and genetic marker of integron and transposon in multi-resistant Pseudomonas aeruginosa(MRPA) isolated from clinical specimens and study their relationship by phylogenetic analysis.METHODS Twenty-one resistant genes,two integron-Ⅰ genes and one transposon genetic gene were analyzed by PCR and verified by DNA sequencing.Multi-resistant genes cluster analysis was performed by UPGMA.RESULTS The positive rates of CARB,oprD2,aac(3)-Ⅱ,aac(6′)-Ⅱ,ant(2″)-Ⅰ,intⅠ1,qacE△1-sul1 and merA in 20 strains of MRPA were 15%,100%,70%,15%,15%,85%,85% and 85%,respectively,and other genes were negative.It was classfied to three subgroup,by the multi-resistant genes cluster analysis.CONCLUSIONS MRPA isolated from clinical specimens has carried many resistant genes.The deficiency rate of oprD2 gene is very high.The positive rate of genetic mark genes about integron and transposon is very high.It may be the main multi-resistant mechanism of MRPA.Multi-resistant genes cluster analysis shows that there is clone transmission in MRPA and it can induce nosocomial infection prevalence.

Antimicrobial-resistant Genes and Consanguinity in Multi-resistant Pseudomonas aeruginosa / 中华医院感染学杂志

OBJECTIVE To investigate the antimicrobial-resistant genes and consanguinity in multi-resistant Pseudomonas aeruginosa(MRPA) isolated from the 98th Hospital of PLA,Huzhou,Zhejiang Province,China.METHODS Thirty strains of MRPA were isolated from hospitalized patients between Sep 2003 and Oct 2004.Twenty four kinds of genes of blaTEM,blaSHV,blaOXA-10 group,blaPER,blaVEB,blaIMP,blaVIM,blaGES,blaCARB,blaDHA,blaMIR,aac(3)-Ⅰ,aac(3)-Ⅱ,aac(3)-Ⅲ,aac(3)-Ⅳ,aac(6′)-Ⅰ,aac(6′)-Ⅱ,ant(3″)-Ⅰ,ant(2″)-Ⅰ,aph(3′)-Ⅵ,oprD,qacE△1-sul1,catB,and cml1 were analyzed by PCR and verified by DNA sequencing.Resistant-genes cluster analysis was performed by Average.RESULTS In 30 strains of MRPA the positive rate of genes of blaTEM,blaOXA-10 group,blaCARB,aac(3)-Ⅱ,aac(6′)-Ⅰ,aac(6′)-Ⅱ,ant(3″)-Ⅰ,ant(2″)-Ⅰ,qacE△1-sul1,and cml1 were 66.7%,3.3%,3.3%,76.7%,3.3%,33.3%,53.3%,26.7%,83.3%,and 3.3%,respectively,and the deficiency rate of oprD gene was 90.0%.The gene of blaOXA-10 group was sequenced and determined to be blaOXA-10 subtype ESBL gene.But the rest of genes were all negative.According to the cluster analysis of resistant-gene,30 strains of MRPA isolated could be classified into four subgroups,which were caused by the infection in hospital.CONCLUSIONS At least 10 kinds of antimicrobial-resistant genes exist in MRPA isolates,and the deficiency rate of oprD gene is very high.MRPA can induce clone transmitted hospital infection and it has fulminant prevalence.