Detection of the Production of Klebsiella Pneumoniae Carbapenemase, New Delhi Metallo-Beta-Lactamase and Oxacillinase-48-Type Carbapenemases by Gram-Negative Bacilli in Resource-Limited Setting

Background: The increasing resistance of bacteria to various antibiotics is a worldwide public health issue. Carbapenems that have elicited great hope in treating infections caused by multidrug-resistant germs have seen their efficacy narrowed over time with the emergence of other novel resistance mechanisms, notably the production of Carbapenemases. Methods: A prospective cross-sectional study was conducted from May 2017 to May 2018 in Douala (Cameroon) to detect carbapenemase-producing Gram-negative bacilli. Isolated strains were identified using the Vitek2TM system. Antimicrobial susceptibility testing was performed using the Kirby-Bauer disk diffusion method on agar plates with 20 selected commercially available antibiotic discs. The bacterial strains were tested for the production of three Carbapenemases (OXA-48, NDM, KPC), using an immuno-chromatographic technique, with the “RESIST-3 O.K.N. K-SeT” rapid detection kit. Results: During the study period, 1687 strains of Gram-negative bacilli were isolated in selected laboratories with a total of 200 multi-resistant strains identified (11.9%). Among the multi-resistant strains, E. coli was the species most represented in Enterobacteriaceae (27.5%) followed by K. pneumoniae (15.5%) and the non-fermenting Gram-negative bacilli were predominantly P. aeruginosa (20.5%). These strains mainly came from urine and pus, i.e. 41% and 32% respectively. Thirty-two (16%) strains produced one of the Carbapenemases with a higher frequency at the General Hospital (84%). NDM-type carbapenemase was the most frequently identified (8.5%), OXA-48 type 7.5%, and no KPC production was observed. Among the Enterobacteriaceae 22.9% produced Carbapenemases and only 5.1% of the non-fermenting bacilli produced these enzymes. The isolates strains were completely resistant to all antibiotics except Amikacin and Fosfomycin. The strains producing the NDM-type carbapenemase showed higher rates of resistance to almost all of the antibiotics tested. Conclusion: Multidrug-resistant strains are experiencing an increase in evolution. The apparition of strains producing Carbapenemases prominently, the NDM and OXA-48 favor this increase. The activities of antibiotics with high efficacies on these strains are low.

Detection of 16S rRNA Methylase Genes in Gram-Negative Bacilli Isolated from Hospitals in Changchun, China

Methylation of 16S rRNA is an important mechanism of aminoglycoside resistance among gram-negative pathogens. In this report, 16S rRNA methylase genes were amplified using PCR among gram-negative bacillus isolates from hospitals in the Changchun area of China and 16S rRNA methylase genotypes (armA, rmtB, rmtA, rmtC, rmtD, and npmA) were identified by direct sequencing. Fifty of the isolates (43.1%) harbored 16S rRNA methylase genes. The common 16S rRNA methylase genes were armA and rmtB (12.1% and 31.0%, respectively), whereas the rmtA, rmtC, rmtD, and npmA genes were absent from the sample. It suggests that the predominant 16S rRNA methylase genes among gramnegative bacilli in the Changchun area are armA and rmtB.

Immunogenomics for identification of disease resistance genes in pigs: a review focusing on Gram-negative bacilli

Over the past years, infectious disease has caused enormous economic loss in pig industry. Among the pathogens, gram negative bacteria not only cause inflammation, but also cause different diseases and make the pigs more susceptible to virus infection. Vaccination, medication and elimination of sick pigs are major strategies of controlling disease. Genetic methods, such as selection of disease resistance in the pig, have not been widely used. Recently, the completion of the porcine whole genome sequencing has provided powerful tools to identify the genome regions that harboring genes controlling disease or immunity. Immunogenornics, which combines DNA variations, transcriptorne, immune response, and QTL mapping data to illustrate the interactions between pathogen and host immune system, will be an effective genomics tool for identification of disease resistance genes in pigs. These genes will be potential targets for disease resistance in breeding programs. This paper reviewed the progress of disease resistance study in the pig focusing on Gram-negative bacilli. Major porcine Gram-negative bacilli and diseases, suggested candidate genes/pathways against porcine Gram-negative bacilli, and distributions of QTLs for immune capacity on pig chromosomes were summarized. Some tools for immunogenomics research were described. We conclude that integration of sequencing, whole genome associations, functional genomics studies, and immune response information is necessary to illustrate molecular mechanisms and key genes in disease resistance.

Low-Level Antibiotic Resistance among Staphylococcus aureus and Gram-Negative Pathogens from Infected Skin and Soft Tissues in Rural Kenya

Introduction: Bacterial skin and soft tissue infections (SSTIs) are a cause of frequent inpatient and outpatient care visits whose causative agents are associated with a high antimicrobial resistance burden. For insights on antimicrobial susceptibilities in a rural setting, we examined specimens from suspected SSTIs from two public health facilities in Kenya. We additionally assessed antibiotic use, appropriateness of empiric therapy and risk factors for SSTI. Methodology: Between 2021 and 2023, 265 patients at Kisii and Nyamira County Referral hospitals were enrolled. Wound swabs/aspirates were collected and processed following standard microbiological procedures. Identification and antimicrobial susceptibility were performed using the VITEK 2 Compact platform. Demographic, clinical, and microbiological data were analyzed with R Statistical software. Results: S. aureus was isolated in 16.2% (43/265) of patients with a methicillin resistance (MRSA) proportion of 14% (6/43). While 13/15 drugs elicited susceptibilities ranging from 84% - 100%, penicillin (16%) and trimethoprim-sulfamethoxazole [TMP-SXT] (23%) yielded the lowest susceptibilities. Escherichia coli (n = 33), Klebsiella pneumoniae (n = 8), Pseudomonas aeruginosa (n = 8), and Citrobacter species (n = 4) were the most commonly isolated gram-negative species. Gram-negative strains showed high susceptibilities to most of the tested drugs (71% - 100%) with the exception of ampicillin (18%), TMP-SXT (33%), and first and second generation cephalosporins. Conclusions: The low MRSA prevalence and generally high antibiotic susceptibilities for S. aureus and gram-negative bacteria present opportunities for antibiotic stewardship in the study setting. Diminished susceptibilities against penicillin/ampicillin and TMP-SXT accord with prevailing local data and add a layer of evidence for their cautious empiric use.

High mortality associated with gram-negative bacterial bloodstream infection in liver transplant recipients undergoing immunosuppression reduction

BACKGROUND Immunosuppression is an important factor in the incidence of infections in transplant recipient.Few studies are available on the management of immunosuppression(IS)treatment in the liver transplant(LT)recipients complicated with infection.The aim of this study is to describe our experience in the management of IS treatment during bacterial bloodstream infection(BSI)in LT recipients and assess the effect of temporary IS withdrawal on 30 d mortality of recipients presenting with severe infection.AIM To assess the effect of temporary IS withdrawal on 30 d mortality of LT recipients presenting with severe infection.METHODS A retrospective study was conducted with patients diagnosed with BSI after LT in the Department of Liver Surgery,Renji Hospital from January 1,2016 through December 31,2017.All recipients diagnosed with BSI after LT were included.Univariate and multivariate Cox regression analysis of risk factors for 30 d mortality was conducted in the LT recipients with Gram-negative bacterial(GNB)infection.RESULTS Seventy-four episodes of BSI were identified in 70 LT recipients,including 45 episodes of Gram-positive bacterial(GPB)infections in 42 patients and 29 episodes of GNB infections in 28 patients.Overall,IS reduction(at least 50%dose reduction or cessation of one or more immunosuppressive agent)was made in 28(41.2%)cases,specifically,in 5(11.9%)cases with GPB infections and 23(82.1%)cases with GNB infections.The 180 d all-cause mortality rate was 18.5%(13/70).The mortality rate in GNB group(39.3%,11/28)was significantly higher than that in GPB group(4.8%,2/42)(P=0.001).All the deaths in GNB group were attributed to worsening infection secondary to IS withdrawal,but the deaths in GPB group were all due to graft-versus-host disease.GNB group was associated with significantly higher incidence of intra-abdominal infection,IS reduction,and complete IS withdrawal than GPB group(P<0.05).Cox regression showed that rejection(adjusted hazard ratio 7.021,P=0.001)and complete IS withdrawal(adjusted hazard ratio 12.65,P=0.019)were independent risk factors for 30 d mortality in patients with GNB infections after LT.CONCLUSION IS reduction is more frequently associated with GNB infection than GPB infection in LT recipients.Complete IS withdrawal should be cautious due to increased risk of mortality in LT recipients complicated with BSI.

Association between infections caused by multidrug-resistant gram-negative bacteria and mortality in critically ill patients

The incidence of gram-negative multidrug-resistant(MDR) bacterial pathogens is increasing in hospitals and particularly in the intensive care unit(ICU) setting. The clinical consequences of infections caused by MDR pathogens remain controversial. The purpose of this review is to summarize the available data concerning the impact of these infections on mortality in ICU patients. Twenty-four studies, conducted exclusively in ICU patients, were identified through Pub Med search over the years 2000-2015. Bloodstream infection was the only infection examined in eight studies, respiratory infections in four and variable infections in others. Comparative data on the appropriateness of empirical antibiotic treatment were provided by only seven studies. In ten studies the presence of antimicrobial resistance was not associated with increased mortality; on the contrary, in other studies a significant impact of antibiotic resistance on mortality was found, though, sometimes, mediated by inappropriate antimicrobial treatment. Therefore, a direct association between infections due to gram-negative MDR bacteria and mortality in ICU patients cannot be confirmed. Sample size, presence of multiple confounders and other methodological issues may influence the results. These data support the need for further studies to elucidate the real impact of infections caused by resistant bacteria in ICU patients.

Seafood as a Reservoir of Gram-negative Bacteria Carrying Integrons and Antimicrobial Resistance Genes in Japan

PCR and DNA sequencing were used to screen and characterize integrons and resistance genes in Gram-negative bacteria isolated from seafood products in Japan.A total of 215 Gram-negative bacteria were isolated from local and imported seafood samples collected from retail markets in Hiroshima Prefecture.Class 1 integrons containing gene cassettes encoding resistance to trimethoprim

Impact of Polymyxin Resistance on Virulence and Fitness among Clinically Important Gram-Negative Bacteria

Humanity is facing an enormous and growing worldwide threat from the emergence of multi-drug-resistant(MDR)Gram-negative bacteria such as Escherichia coli,Klebsiella pneumoniae,and Acinetobacter baumannii.Polymyxin B and E(colistin)constitute the last-line therapies for treating MDR Gram-negative bacteria.Polymyxin is a cationic antibacterial peptide that can destroy the outer membrane of Gram-negative bacteria.With the increasing clinical application of polymyxin,however,there have been many reports of the occurrence of polymyxin-resistant Gram-negative bacteria.This resistance is mainly mediated by the modification or complete loss of lipopolysaccharide(LPS).LPS is also a virulence factor of Gram-negative bacteria,and alterations of LPS may correlate with virulence.Although it is generally believed that the biological costs associated with drug resistance may enable benign susceptible bacteria to overcome resistant bacteria when antibiotic pressure is reduced,some studies have shown that polymyxin-resistant bacteria are associated with higher virulence and greater fitness compared with their susceptible counterparts.To predict the development of polymyxin resis-tance and evaluate interventions for its mitigation,it is important to understand the relative biological cost of polymyxin resistance compared with susceptibility.The impact of polymyxin resistance mecha-nisms on the virulence and fitness of these three Gram-negative bacteria are summarized in this review.

An Improved Approach for Rapidly Identifying Different Types of Gram-Negative Bacterial Secreted Proteins

Protein secretion plays an important role in bacterial lifestyles. In Gram-negative bacteria, a wide range of proteins are secreted to modulate the interactions of bacteria with their environments and other bacteria via various secretion systems. These proteins are essential for the virulence of bacteria, so it is crucial to study them for the pathogenesis of diseases and the development of drugs. Using amino acid composition (AAC), position-specific scoring matrix (PSSM) and N-terminal signal peptides, two different substitution models are firstly constructed to transform protein sequences into numerical vectors. Then, based on support vector machine (SVM) and the “one to one”?algorithm, a hybrid multi-classifier named SecretP v.2.2 is proposed to rapidly and accurately?distinguish different types of Gram-negative?bacterial secreted proteins. When performed on the same test set for a comparison with other methods, SecretP v.2.2 gets the highest total sensitivity of 93.60%. A public independent dataset is used to further test the power of SecretP v.2.2 for predicting NCSPs, it also yields satisfactory results.

Metallo-β-lactamase producing nonfermentative gram-negative bacteria:An increasing clinical threat among hospitalized patients

Objective:To detect and evaluate the various methods for metallo-β-lactamases(MBL) production in Pseudomonas aeruginosa(P.aeruginosa) and Acinetobacter species.Methods:A total of 109 P.aeruginosa and 85 Acinetobacter species were screened for imipenem resistance by Kirby- Bauer disc diffusion methods.Detection of MBL production was(lone by imipenem-EDTA combined disc test,double disc synerygy test(DDST) and imipenem-EDTA MBL E test.Results: A total of 63(57.8%) strains of P.aeruginosa and 46(54.1%) strains of Acinetobacter spp.were found to be resistant to imipenem.Of the 63 imipenem resistant P.aeruginosa tested for MBL production.44(69.89;) were found to be positive and among 46 imipenem resistant Acinetobacter. 19(41.3%) were shown to be the MBL producers.Conclusions:Imipenem-EDTA combined disc test and MBL E test are equally effective for MBL detection in both P.aeruginosa and Acinetobacter spp.,but given the cost-constraints,combined disc can be used as a convenient screening method in the clinical microbiology laboratory.

Epidemiological Characteristics, Resistance Patterns and Spread of Gram-Negative Bacteria Related to Colonization of Patients in Intensive Care Units

Our aim was to determine the epidemiological characteristics, the resistance patterns and the spread of Gram negative bacteria related to colonization of patients in adult Intensive Care Units. Methods: A prospective cohort of patients colonized and/or infected with Gram negative bacteria was conducted at two adult ICUs from hospitals in Brazil (April 2012 to February 2013). Nasal, groin and perineum swabs were performed. Samples were incubated on MacConkey and cetrimide agar (48 h at 37℃) and identification tests (Vitek-BioMérieux), antibiogram (Bauer-Kirby method), Carba NP test, Polymerase Chain Reaction (PCR) and sequencing were performed. The patterns of resistant microorganisms were compared by rep-PCR (Diversilab). Results: There were 53 cases of colonization. In these cases, we identified imipenem-resistant Acinetobacter baumannii (51%), Pseudomonas aeruginosa (32%), Klebsiella pneumoniae ESBL (38%) or imipenem resistant (5.6%). The use of antimicrobials and medical devices were related to colonization (p The resistance patterns expressed by Klebsiella pneumoniae were ESBL (CTX-M, SHV e TEM) and KPC2. A verified profile of Acinetobacter baumannii was related to OXA-23 and OXA-253 (OXA-143 variant). The profiles ESBL and KPC2 expressed by Klebsiella pneumoniae were distributed between the both ICUs. The distribution of OXA-23 and OXA-253 was verified only in one ICU. The similarity of strains ranged from 80% to 95%, highlighting the horizontal transference of these microorganisms.

Prevalence and Antimicrobial Resistance of Gram-Negative Bacteria Isolates in Shellfish Samples from Two River Estuaries in South-South Nigeria

Antibiotic resistant bacteria pathogens remain the leading cause of shellfish borne diseases and a major health threat to humans worldwide. The objectives of this study were to isolate, identify, and determine the antibiotic resistance patterns of Gram-negative bacteria from shellfish. We analyzed a total of 540 shellfish (117 clams, 88 oysters, and 136 periwinkles) samples collected from different vendors at Iko and Douglas Creeks in Akwa Ibom State, South-South Nigeria. Conventional cultural techniques, morphological, biochemical characteristics, and PCR amplification were used to identify the bacterial isolates. Antibiotic susceptibility tests (Kirby-Bauer disk diffusion method) and ESBL phenotype (disk) of the isolates were performed. One hundred and thirty-five (135) Gram-negative bacteria comprising 5 genera and 14 species were detected at a prevalence of: Alcaligenes faecalis TRB-7 38 (28.2%), Pseudomonas oryzihabitans strain KCB005 16 (11.9%), Paenalcaligenes retgerii strain B5 12 (8.9%) Pseudomonas aeruginosa JB2 10 (7.4%), Providencia stuartii DMC-28b 9 (6.7%), Alcaligenes species TLT151 8 (5.9%), Pseudomonas aeruginosa CIFRI DTSB1 7 (5.2%), Paenalcaligenes species UN24 7 (5.2%), Alcaligenes faecalis BT10 7 (5.2%), Vibrio species strain PrVy108 6 (4.4%), Pseudomonas xiamenensis C10-2 5 (3.7%), Providencia vemicola Bu15_38 4 (2.9%), Pseudomonas anguillisceptica 4029 3 (2.2%), and Pseudomonas aeruginosa N15-01092 3 (2.2%). All tested isolates showed various degrees of resistance to the thirteen antimicrobials evaluated. High levels of resistance (100%) to cefepime and imipenem were expressed by all isolates except the Providencia species. For the EBSL indicators, all isolates apart from Alcaligenes species were resistant (100%) to ceftriaxone. All Vibrio species were susceptible to norfloxacin, nalidixic acid, and ceftazidime. The identification of antibiotic resistant Gram-negative bacteria (GNARB) from shellfish in this study highlights the risk of disseminated multi-drug resistance—a serious public health concern.

Impact of an Antimicrobial Stewardship Program Initiative on Antibiotic Optimization in Patients with Gram-Negative Bacteremia

The purpose of this study was to critically evaluate the impact of an institutional blood culture notification protocol called RAIDS （rapid administration of antimicrobials by an infectious diseases specialist） on time to optimization of antimicrobial therapy in hospitalized patients with gram-negative bacteremia. Time to antibiotic optimization was compared in patients with gram-negative bacilli isolated from blood cultures obtained from March-May 2011 （pre-RAIDS） versus March-May 2013 （post-RAIDS）. The results show that patients in the pre-RAIDS study group had a significantly longer time to antibiotic optimization when compared to the post-RAIDS group （median （IQR）, 27.6 （10.8-75.8） h vs. 3.1 （0.8-34.3） h, p = 0.03）. The RAIDS protocol resulted in quicker time to antibiotic de-escalation （pre- vs. post-RAIDS; median （IQR）, 27.6 （10.8-134.5） h vs. 4.3 （1.4-32.6） h, p = 0.03）. There were no differences in clinical outcomes such as clinical cure, microbiological cure, and 30-day mortality between pre-RAIDS and post-RAIDS study groups. Patients in the post-RAIDS arm were more likely to receive appropriate empiric and definitive treatment. Implementation of the RAIDS protocol, which was an ASP （antimicrobial stewardship program） initiative, resulted in quicker time to antibiotic de-escalation and overall treatment optimization. RAIDS reduced the unnecessary use of broad-spectrum antimicrobial in this study population.

Comparison of the serum contents of inflammatory mediators and oxidative stress mediators between patients with gram-positive bacteria and gram-negative bacteria infection

Objective: To compare the serum contents of inflammatory mediators and oxidative stress mediators between patients with gram-positive bacteria and gram-negative bacteria infection. Methods: Patients who were diagnosed with bloodstream bacterial infection in Zigong Third People's Hospital between March 2015 and April 2017 were selected as the research subjects and divided into gram-positive group and gram-negative group according to the results of blood culture and strain identification, and serum levels of inflammatory mediators PCT, IL-1β, IL-6, sTREM-1, TNF-α, NGAL, SAA, HPT and hs-CRP as well as oxidative stress mediators MDA, AOPP, TAC, CAT and SOD were determined. Results: Serum PCT, IL-1β, IL-6, sTREM-1, TNF-α, NGAL, SAA, HPT, hs-CRP, MDA and AOPP levels of gram-negative group were greatly higher than those of gram-positive group while TAC, CAT and SOD levels were greatly lower than those of gram-positive group. Conclusion: The changes of inflammatory mediators and oxidative stress mediators in the serum of patients with gram-negative bacteria infection are more significant than those of patients with gram-positive bacteria infection.

Diagnostic efficacy of Ziehl-Neelsen method against fluorescent microscopy in detection of acid fast bacilli

Objective:To investigate the application of Ziehl-Neelsen(Z-N) and fluorescent microscopy in detection of acid fast bacilli(AFB).Methods:Duplicate smears were prepared from 260 sputum samples and stained with Z-N and fluorescent staining(FS) methods.The efficiency of both methods in primary diagnosis of tuberculosis were evaluated.Results:The smears were positive for AFB in 15(5.77%) samples by Z-N staining method and in 16(6.15%) samples by FS method.The sensitivity and specificity of Z-N staining method against FS method were 93.75% and 100%respectively.Conclusions:Though lesser cost-effective than Z-N,FS method is a more sensitive and better case finding tool in detection of AFB.

Experimental Study on the Antibacterial Effect of Origanum Volatile Oilon Dysentery Bacilli In Vivo and In Vitro

To observe the germistatic and germicidal effects of origanum volatile oil (OVI) on the dysentery bacteria, the abdominal cavity of mice was infected with Shigella sonne (Sh. sonnei) and Shigella flexneri (Sh. flexneri) F 2a. After OVI was given to the mice via gastric lavage, the effects of OVI on the infected mice were observed. The minimal inhibitory concentration (MIC) and minimal bactericidal concentration (MBC) for dysentery bacteria were determined in vitro. The results showed that origanum volatile oil showed obvious protective effect on mice infected with Sh. sonnei and Sh. flexneri F 2a. and it had germistatic and germicidal effects on dysentry bacteria. We are led to conclude that origanum volatile oil is an effective medicine against the infection of dysentery bacteria.

Detection of Acid Fast Bacilli in Tuberculous Lymph Node Tissue and Touch Preparations—A Comparative Study

Background: The most common form of extrapulmonary tuberculosis is lymphadenitis. Lymphadenitis is considered mainly a disease of young children and has a peak age of onset at 20 to 40 years. Objectives: The purpose of this study was to detect and compare the presence of acid fast bacilli in lymph node biopsies and their touch preparations using Ziehl Neelsen and fluorescence staining. Materials and Methods: The lymph node biopsies and their touch preparations were prepared and stained with Haematoxylin and eosin to determine their morphology. Slides were stained with ZN staining and fluorescent staining technique using Auramine O and Rohdamine B to demonstrate the presence of acid fast bacilli in tissue and their touch preparations. Results: AFB was detected in 56% of biopsies and 54% of touch preparations. In addition in these cases we observed that fluorescent staining technique using Auramine O and Rohdamine B is superior to ZN stain because low magnification used in fluorescent staining technique makes it possible to scan the smear rapidly and detects the AFB as glowing spots even if it is present in small numbers. Conclusion: The results of AFB in tissue biopsies are in equivalent in their touch preparation.

Detection of oqxA and oqxB efflux pump genes among nosocomial coliform bacilli:An observational cross-sectional study

Objectives:To identify and test the antibiotic susceptibility of nosocomial coliform bacilli and investigate the presence of oqxA and oqxB genes among the multidrug-resistant(MDR)phenotypes.Methods:One hundred and twenty different healthcare-associated infection samples were collected.Coliform bacilli were isolated,identified by conventional methods,and then antibiotic susceptibility tests were done using the VITEK2 system and disk diffusion methods.OqxAB operon was identified using a conventional PCR-based technique.oqxA and oqxB genes were compared between MDR Klebsiella pneumonia(K.pneumonia)phenotypes and MDR Escherichia coli(E.coli)phenotypes.Besides,oqxAB operons were compared between phenotypes of K.pneumonia and E.coli isolates.Results:Seventy coliform bacilli were isolated with the predominance of K.pneumonia and E.coli.Besides,82.1%of K.pneumonia strains and 53.3%of E.coli isolates were MDR phenotypes.Significant more oqxB genes alone were found in MDR E.coli than that in MDR K.pneumoniae phenotypes(χ^(2)=10.160,P=0.003).OqxAB operon was significantly more in MDR phenotypes of E.coli than that in the susceptible phenotypes(P<0.001).There was significantly less of this operon in susceptible E.coli isolates than that in susceptible K.pneumoniae isolates(P<0.001).OqxAB positive isolates that were also resistant to fluoroquinolones,tetracycline,trimethoprim,and chloramphenicol,most probably suggested functional pumps.Conclusions:MDR coliform bacilli are strongly implicated in healthcare-associated infection.Attention should be paid to the presence of oqxAB pump,as an important mechanism in the development of resistance against many antimicrobials because it contributes to co-resistance with other categories;therefore,this pump could be a good target for efflux pump inhibitors.

Obligate aerobic, gram-positive, weak acid-fast, nonmotile bacilli, Tsukamurella tyrosinosolvens: Minireview of a rare opportunistic pathogen

Tsukamurella species are obligate aerobic,gram-positive,weak acid-fast,nonmotile bacilli.They are found in various environments,such as soil,water,sludge,and petroleum reservoir wastewater,and belong to the order Actinomycetales.In 2016,there was a reclassification of species within the genus Tsukamurella,merging the species Tsukamurella tyrosinosolvens(T.tyrosinosolvens)and Tsukamurella carboxydivorans.Tsukamurella species are clinically considered to be a rare opportunistic pathogen,because most reported cases have been related to bacteremia and intravascular prosthetic devices and immunosuppression.To date,it has been isolated only from human specimens,and has always been associated with clinical disease;human infections are very rare.Reported infections have included pneumonia,brain abscesses,catheter-related bloodstream infections,ocular infections,bacteremia,and sepsis presenting with septic pulmonary emboli in patients who are immunocompromised.To date,there is no commercially available test for identification.On the other hand,sequence-based identification,including matrix-assisted laser desorption ionization time-of-flight mass spectrometry,is an alternative method for identifying clinical isolates that are either slow growers or difficult to identify through biochemical profiling.The golden standards for diagnosis and optimal management still remain to be determined.However,newer molecular biological techniques can provide accurate identification,and contribute to the appropriate selection of definitive therapy for infections caused by this organism.Combinations of several antimicrobial agents have been proposed for treatment,though the length of treatment for infections has yet to be determined,and should be individualized according to clinical response.Immunocompromised patients often experience severe cases due to infection,and life-threatening T.tyrosinosolvens events associated with dissemination and/or failure of source control have occurred.Favorable prognoses can be achieved through earlier identification of the cause of infection,as well as successful management,including appropriate antibiotic therapy together with source control.Further analyses of similar cases are required to establish the most adequate diagnostic methods and treatment regimens for infections.

2015—2021年CHINET儿童患者临床分离不发酵糖革兰阴性杆菌耐药性变迁

目的了解CHINET中国细菌耐药性监测网儿童患者临床分离的不发酵糖革兰阴性杆菌的分布和耐药性变迁。方法按CHINET耐药监测统一方案使用纸片扩散法或商品化药敏试验自动测试仪进行药敏试验。按CLSI 2021年版标准判断结果。结果临床分离不发酵糖革兰阴性杆菌26987株中,主要以假单胞菌属和不动杆菌属细菌为主,其他各菌属均有不等检出。下呼吸道标本占51.1%,居于首位,脓液及伤口分泌物占16.5%,尿液标本占9.2%。这些菌株主要分离于内科病房(38.1%)、儿科ICU(26.8%)和外科病房(18.9%)。铜绿假单胞菌对氨曲南耐药率最高,为22.7%,对碳青霉烯类抗生素耐药率为15.6%,对其他抗菌药物耐药率均低于10%。碳青霉烯类耐药鲍曼不动杆菌检出率54.1%,明显高于铜绿假单胞菌的15.6%,2020年和2021年出现下降趋势。碳青霉烯类耐药铜绿假单胞菌在新生儿期患儿的检出率(34.7%)高于其他患儿;碳青霉烯类耐药鲍曼不动杆菌随患儿年龄增长检出率逐渐增高(39.9%~65.4%)。结论儿童分离铜绿假单胞菌和鲍曼不动杆菌对各种抗菌药物的耐药率呈锯齿形变化,对大多抗菌药物的耐药率有下降趋势。不同年龄段患儿中碳青霉烯类耐药菌株的检出率不同。不发酵糖革兰阴性杆菌的分布特征和不同菌属菌株的耐药特征存在差异。因此,在儿科对此类菌进行耐药监测十分必要。