Background: The purpose of this study was to analyze the relationship between the gene mutation patterns by the GenoType MTBDRplus (MTBDRplus) assay and the phenotypic drug susceptibility test (pDST) results of isoniazid (INH) and prothionamide (Pto). Methods: A total of 206 patients whose MTBDRplus assay results revealed katG or inhA mutations were enrolled in the study. The pDST results were compared to mutation patterns on the MTBDRplus assay. Results: The katG and inhA mutations were identified in 68.0% and 35.0% of patients, respectively. Among the 134 isolated katG mutations, three (2.2%), 127 (94.8%) and 11 (8.2%) were phenotypically resistant to low-level INH, high-level INH, and Pto, respectively. Among the 66 isolated inhA mutations, 34 (51.5%), 18 (27.3%) and 21 (31.8%) were phenotypically resistant to low-level INH, high-level INH, and Pto, respectively. Of the 34 phenotypic Pto resistant isolates, 21 (61.8%), 11 (32.4%), and two (5.9%) had inhA, katG, and both gene mutations. Conclusion: It is noted that Pto may still be selected as one of the appropriate multidrug-resistant tuberculosis regimen, although inhA mutation is detected by the MTBDRplus assay until pDST confirms a Pto resistance. The reporting of detailed mutation patterns of the MTBDRplus assay may be important for clinical practice, rather than simply presenting resistance or susceptibility test results.
Salman, Hamzah Abdulrahman;Alhameedawi, Alaa kamil;Alsallameh, Sarah Mohammed Saeed;Muhamad, Ghofran;Taha, Zahraa
Microbiology and Biotechnology Letters
/
v.50
no.1
/
pp.147-156
/
2022
Urinary tract infections (UTIs) are one of the most common infections in different age groups, including children. Bacteria are the main etiological agents of UTIs. The aim of the present study was to isolate, identify, and determine the antibiotic susceptibility of bacteria isolated from children with UTIs from Baghdad, Iraq. Three hundred and two urine samples were collected from children aged 6 months to 12 years. The samples were cultured on blood agar and MacConkey agar. The selected colonies were subjected to biochemical tests and antibiotic susceptibility analysis using the Vitek® 2 Compact automated microbial identification system. In this sample, 299 bacteria were identified, of which, 267 were gram-negative bacteria, and 32 were gram-positive bacteria. Escherichia coli (56%) was the most commonly isolated gram-negative bacteria, followed by Pseudomonas aeruginosa (14%), Enterobacter spp. (10.48%), Klebsiella pneumoniae (9.36%), Proteus spp. (7.8%), Acinetobacter baumannii (1.5%), and Morganella morganii (0.37%). Enterococcus faecalis (62.5%) was the most commonly detected gram-positive bacteria, followed by Staphylococcus aureus (37.5%). E. coli and P. aeruginosa were the most antibiotic-resistant bacteria. Among the tested antibiotics, meropenem showed 100% sensitivity, followed by imipenem (97.4%), amikacin (91.8%), and tobramycin (83.5%). In contrast, the high frequencies of resistance were observed with cefixime (93.2%), cefotaxime (78.7%), and ceftriaxone/cefotaxime (71.2%). In conclusion, carbapenems and aminoglycosides are highly recommended for the empirical treatment of UTIs, while, Quinolones, penicillins, and cephalosporins are not suggested. Frequent antibiotics susceptibility testing are warranted to determine the resistance pattern of UTI bacteria.
We investigated the virulence genes, O-serotypes, antimicrobial resistance of pathogenic E. coli isolated from carcasses (n=455) and environmental (n=372) samples of 11 cattle and 12 pig slaughterhouses from December 2020 to December 2021. E. coli were isolated from nine carcasses (2.0%), three slaughter facilities (1.4%), two utensils (2.7%) and three abattoir workers (3.5%) from four cattle and four pig slaughterhouses. Among all isolates, 13 STEC (76.5%) were identified, followed by four EPEC (23.5%). As a result of the antibiotic susceptibility test, all isolates were resistant to at least one antimicrobial, of which 70.6% isolates showed multidrug resistance patterns. The serotypes were diverse in pigs compared to cattle, with serotypes O18, O66, O109 in cattle and O9, O76, O85, O100, O153, and O159 in pigs. In a single cattle slaughterhouse, eight STEC O66 were isolated from various types of sample (4 slaughter animal surfaces, 3 gloves, and 1 knife) with two antimicrobial resistance patterns (CHL-FIS-STR and CHL-FIS). Those two types of strain were suspected cross-contamination from utensils to slaughter animal surfaces. These results showed that pathogenic E. coli were detected in carcasses and various environmental samples in cattle and pig slaughterhouses. Nationwide monitoring and hygiene management are required to prevent cross-contamination of STEC isolate slaughterhouses.
As the field of interventional pain management (IPM) grows, the risk of surgical site infections (SSIs) is increasing. SSI is defined as an infection of the incision or organ/space that occurs within one month after operation or three months after implantation. It is also common to find patients with suspected infection in an outpatient clinic. The most frequent IPM procedures are performed in the spine. Even though primary pyogenic spondylodiscitis via hematogenous spread is the most common type among spinal infections, secondary spinal infections from direct inoculation should be monitored after IPM procedures. Various preventive guidelines for SSI have been published. Cefazolin, followed by vancomycin, is the most commonly used surgical antibiotic prophylaxis in IPM. Diagnosis of SSI is confirmed by purulent discharge, isolation of causative organisms, pain/tenderness, swelling, redness, or heat, or diagnosis by a surgeon or attending physician. Inflammatory markers include traditional (C-reactive protein, erythrocyte sedimentation rate, and white blood cell count) and novel (procalcitonin, serum amyloid A, and presepsin) markers. Empirical antibiotic therapy is defined as the initial administration of antibiotics within at least 24 hours prior to the results of blood culture and antibiotic susceptibility testing. Definitive antibiotic therapy is initiated based on the above culture and testing. Combination antibiotic therapy for multidrug-resistant Gram-negative bacteria infections appears to be superior to monotherapy in mortality with the risk of increasing antibiotic resistance rates. The never-ending war between bacterial resistance and new antibiotics is continuing. This article reviews prevention, diagnosis, and treatment of infection in pain medicine.
Jose Roberto Aguirre-Sanchez;Nohemi Castro-del Campo;José Andres Medrano-Felix;Alex Omar Martínez-Torres;Cristobal Chaidez;Jordi Querol-Audi;Nohelia Castro-del Campo
Journal of Veterinary Science
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v.25
no.4
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pp.42.1-42.12
/
2024
Importance: Bovine mastitis, predominantly associated with gram-positive Staphylococcus aureus, poses a significant threat to dairy cows, leading to a decline in milk quality and volume with substantial economic implications. Objective: This study investigated the incidence, virulence, and antibiotic resistance of S. aureus associated with mastitis in dairy cows. Methods: Fifty milk-productive cows underwent a subclinical mastitis diagnosis, and the S. aureus strains were isolated. Genomic DNA extraction, sequencing, and bioinformatic analysis were performed, supplemented by including 124 S. aureus genomes from cows with subclinical mastitis to enhance the overall analysis. Results: The results revealed a 42% prevalence of subclinical mastitis among the cows tested. Genomic analysis identified 26 sequence types (STs) for all isolates, with Mexican STs belonging primarily to CC1 and CC97. The analyzed genomes exhibited multidrug resistance to phenicol, fluoroquinolone, tetracycline, and cephalosporine, which are commonly used as the first line of treatment. Furthermore, a similar genomic virulence repertoire was observed across the genomes, encompassing the genes related to invasion, survival, pathogenesis, and iron uptake. In particular, the toxic shock syndrome toxin (tss-1) was found predominantly in the genomes isolated in this study, posing potential health risks, particularly in children. Conclusion and Relevance: These findings underscore the broad capacity for antibiotic resistance and pathogenicity by S. aureus, compromising the integrity of milk and dairy products. The study emphasizes the need to evaluate the effectiveness of antibiotics in combating S. aureus infections.
This study was conducted to investigate the distribution of Salmonella spp. from pigs and cattle in slaughterhouse, the antimicrobial resistance pattern and the prevalence of resistance genes of isolates. A total of 640 fecal samples from pigs and cattle in slaughterhouse were collected for isolation of Salmonella spp.. Isolation rate was revealed as 15% in pigs and 1.6% in cattle. As result of serotyping, group B (56.6%) were identified as most common in pigs and cattle isolates, in order of group C (24.5%) and group E (15.1%). S. Typhimurium (50.9%) was most common serotype. The major serotypes were in order of S. Rissen and S. London (11.3%) and S. Riggil (7.6%). In antimicrobial test, all isolates were demonstrates susceptibility to nitrofurantoin. But isolates were revealed resistance other antibiotics in order of tetracycline (64.6%), streptomycin (68.3%), ampicillin and amoxicillin (56.3%) and spectinomycin (47.9%). With polymerase chain reaction, antimicrobial resistance gene strA (75.0%) and aadA1 (3.1%) were detected in streptomycin resistance isolates and tetA (94.3%) and tetB (11.3%) gene were detected in tetracycline resistant isolates, but tetG was not detected. Class 1 integron gene was detected in all Salmonella isolates.
Kim Junho;Kim Yeun;Bae Kiho;Song Taek-Sun;Cho Sang-Nae;Lee Hyeyoung
Biomedical Science Letters
/
v.11
no.4
/
pp.465-471
/
2005
Ofloxacin has antimycobacterial activity that possibly contributes a pivotal role in the second-line drug regimens that are used for the treatment of multidrug-resistant tuberculosis. However, in some communities, the resistance rate of Mycobacterium tuberculosis to this agent is surging. Therefore, a rapid and accurate method that can be used to determine the resistance of M tuberculosis to the ofloxacin can be very useful for effective treatment of the patients. As an effort to develop such a method, this study was set up to reveal general types of mutations that are related to ofloxacin resistance of M tuberculosis. From previous studies, it has been well known that ofloxacin resistance is associated with mutations in a gene encoding the gyrase A subunit protein. In this study, we obtained 43 ofloxacin-resistant and 50 ofloxacin-susceptible M tuberculosis clinical isolates from Masan National TB Hospital, and sequences of DNA fragment of 320 bp, region of gyrA corresponding to the ofloxacin resistance-determining region were analyzed. In brief, the results showed that a total of seven mutation types were found at gyrA. Theses mutations were all clustered within nucleotides 2574 to 2586 of the gyrA gene (codons 88 to 94). Codon 94 was the most frequently substituted site. Twenty-four of the 43 isolates had mutations at this position resulting in a total of five different types of amino acid changes $(Asp{\to}Ala,\;Asp{\to}Gly,\;Asp{\to}His,\;Asp{\to}Tyr,\;and\;Asp{\to}Asn)$. Five isolates contained a mutation at codon 90 resulting $Ala{\to}Val$ change. Four isolates had mutations at codon 91 causing a $Ser{\to}Pro$ change at this site. Two isolates contained a mutation at codon 88 and each of them resulted in different types of amino acid changes $(Gly{\to}Cys,\;Gly{\to}Ala)$. On the other hand, polymorphic site at codon 95 was found in both ofloxacin-resistant and ofloxacin-susceptible isolates. From these results, we concluded that the rate of mutations present in gyrA among ofloxacin-resistant M. tuberculosis in Korea is similar to the general rates of mutations found throughout the world. Subsequently, an oligonucleotide probe was designed based on the results of sequence analysis and was used to develop a dot blot hybridization assay system to determine ofloxacin-resistance of M tuberculosis. To evaluate this probe, dot-blot hybridization was carried out using other 57 clinical isolates, and the results showed that the dot-blot hybridization assay is good for detecting sequence alterations atgyrA gene.
The prevalence of multidrug-resistant TB (MDR-TB) and extensively drug-resistant TB (XDR-TB) are increasing. We analyzed the patterns of drug resistance and tracking period days of acquiring anti-mycobacterial resistance. From January 2010 to December 2019, drug susceptibility tests (DST) were performed by the absolute concentration method using the Löwenstein-Jensen solid medium and pyrazinamidase activity test (to assess pyrazinamide resistance) in samples from patients who were referred to the Green Cross Laboratories in Yongin. Among the cases that showed resistance to one or more anti-tuberculosis drugs, 55 patients (33.1%) were resistant to isoniazid (INH) at the time of initial referral, and the rates for the development of resistant anti-tuberculosis drugs were ethambutol (EMB) (26.6%), rifampicin (RFP) (21.9%), quinolones (QUI) (21.9%) and pyrazinamide (PZA) (10.9%), in that order. In the cases sensitive to all 10 anti-tuberculosis drugs initially, the development of resistance to INH was the most frequent, seen in 43 patients (7.2%). The average follow-up period was 435.6 days, and the resistance development was observed in the order of INH (7.2%), RFP (3.9%), SM (1.9%), QUI (0.7%), amikacin (AMK) (0.5%), and EMB (0.5%). The conversion of susceptible strains to resistant strains is an important warning sign for the patient, especially in cases of conversion to MDR or XDR. This information would be helpful for improving patient care during TB treatment.
Kim, Hyoung-Soo;Rho, Kwang-Suk;Kong, Suck-Jun;Sohn, Mal-Hyeun;Kim, Tae-Yoon
Tuberculosis and Respiratory Diseases
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v.51
no.5
/
pp.409-415
/
2001
Background : Multidrug-resistant tuberculosis(MDR-TB) in patients is mainly caused by acquired drug resistance. However, a small proportion of MDR-TB is caused by initial drug resistance(IDR), which may be somewhat different from acquired drug resistance. This study analyzed the clinical characteristics of IDR in MDR -TB patients to use the results as basic data in managing the disease. Methods : A retrospective study of 30 IDR cases in MDR-TB patients from Jan. 1995 to Dec. 1998 was performed. In order to analyze the clinical characteristics, the age, sex, family history, duration of negative conversion, number of resistant drugs, treatment regimens, duration of treatment, extent of disease and cavitary lesion on the chest X-ray was examined. In order to analyze the level of improvement, the extent of the disease and cavitary lesion on the chest X-ray, tested by Wilcoxon signed rank sum test, and the disease free interval rate of 1-year and 4-year was examined using the Kaplan-Meier method. Results : The mean age of the patients was 46.6 years and the sex ratio 1:1. Six(20%) patients had a family history. The mean negative conversion of the sputum AFB stain was 2.6 months. The number of resistant drugs was 7.6 and the number of used drugs 3.6. Twenty-three(67%) patients were treated for less than 12months and 28(93%) patients were treated with first-line drugs. The extent of the disease and the cavitary lesion on the chest X-ray improved after treatment(p<.05). Among 13 patients who were followed up for 22.6 months, 2(15%) patients relapsed and the disease free interval rate of I-year and 4-year was 85%. Conclusion: It is recommended that the duration of treatment of IDR in MDR-TB with first-line drugs be 9-12 months even if the extent of disease and cavitary lesion on the chest X-ray improves.
Among 53 Acinetobacter baumannii isolates collected in 2004, nine imipenem-resistant isolates were obtained from clinical specimens taken from patients hospitalized in Busan, Korea. Nine carbapenemase-producing isolates were further investigated in order to determine the mechanisms underlying resistance. These isolates were then analyzed via antibiotic susceptibility testing, microbiological tests of carbapenemase activity, pI determination, transconjugation test, enterobacterial repetitive consensus (ERIC)-PCR, and DNA sequencing. One outbreak involved seven cases of infection by A. baumannii producing OXA-23 ${\beta}-lactamase$, and was found to have been caused by a single ERIC-PCR clone. During the study period, the other outbreak involved two cases of infection by A. baumannii producing IMP-1 ${\beta}-lactamase$. The two clones, one from each of the outbreaks, were characterized via a modified cloverleaf synergy test and an EDTA-disk synergy test. The isoelectric focusing of the crude bacterial extracts detected nitrocefin-positive bands with pI values of 6.65 (OXA-23) and 9.0 (IMP-1). The PCR amplification and characterization of the amplicons via direct sequencing showed that the clonal isolates harbored $bla_{IMP-1}$ or $bla_{oxA-23}$ determinants. The two clones were characterized by a multidrug resistance phenotype that remained unaltered throughout the outbreak. This resistance encompassed penicillins, extended-spectrum cephalosporins, carbapenems, monobactams, and aminoglycosides. These results appear to show that the imipenem resistance observed among nine Korean A. baumannii isolates could be attributed to the spread of an IMP-lor OXA-23-producing clone. Our microbiological test of carbapenemase activity is a simple method for the screening of clinical isolates producing class D carbapenemase and/or class B $metallo-{\beta}-lactamase$, in order both to determine their clinical impact and to prevent further spread.
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