Antimicrobial Resistance Testing Research Articles

Antibiotic Resistance and Probiotics: Knowledge Gaps, Market Overview and Preliminary Screening.

Probiotics are among those products, the use of which is increasing, and they are available primarily as food/dietary supplements, as well as in the form of medicines. This study aims to assess the attitudes and practices of health professionals and students of health sciences, give a short overview of the probiotics currently on the market, and conduct a screening of five food supplements and one drug with respect to antibiotic resistance. Nearly half of the respondents in our survey state that probiotics have no side effects, while only 6.3% believe that the use of probiotics can lead to antibiotic resistance. In addition, more than 40% of the participants throw unused probiotics into municipal waste. The market analysis results indicate that probiotic products on the Serbian market have highly variable CFU counts, while the declared health claims cover numerous beneficial health effects, and they are sometimes even registered as medicines. Lactobacilli are frequently present in probiotic supplements, and are sold in pharmacies and online. The experimental results showed that antibiotic resistance is present in different types of lactobacilli in probiotic products. The risk of using probiotics, regardless of their beneficial health effects, should be taken into account in the future. An update to the regulations governing probiotics, including a stipulation for antimicrobial resistance (AMR) testing, should be established, and guidelines for their proper use and disposal put into place.

Capacity of laboratories as surveillance sites for Neisseria gonorrhoeae culture and antimicrobial resistance or susceptibility testing in public hospitals within Kisumu County, Kenya

Capacity of laboratories as surveillance sites for Neisseria gonorrhoeae culture and antimicrobial resistance or susceptibility testing in public hospitals within Kisumu County, Kenya

Prevalence, Antimicrobial Drug Resistance and Associated Risk Factors of Streptococcus Pneumoniae Bacteria Infection Among Under-Five Children With Acute Lower Respiratory Tract Infection Attending Sheik Hassan Yebere Referral Hospital, Jig-Jiga, Ethiopia.

Pneumonia is inflammation of the lung. The Streptococcus pneumoniae (S. pneumoniae) is commensal in the upper airway and can cause infection to under-five children. The bacteria is gram-positive diplococci, catalase negative, and optochin sensitive. The bacteria is the leading cause of bacterial pneumonia among under-five children. No similar data is reported from the current study area. To determine prevalence, antimicrobial drug resistance and associated factors of S. pneumoniae infection among under-five children with acute lower respiratory tract infection attending Sheck Hassan Yebere Referral Hospital from March 1 to April 30, 2021 Jig-Jiga, Ethiopia. A cross-sectional study was conducted among 374 study participants selected by convenience sampling method. A structured questionnaire was used to collect child data. Nasopharyngeal/oropharyngeal swabs were collected and diagnosed to isolate S. pneumoniae by using culture then identified by biochemical examination. Later antimicrobial drug resistance testing was performed by Kirby-Bauer disk diffusion method. All data were entered on epi-data 3.1 then exported to SPSS version 22 to calculate analysis. Statistically significant value was found by calculating an adjusted odds ratio with p-value ≤ 0.05 in a multivariate logistic regression model. Among 374 under-five children, 180 (48.1%) were males and 109 (29.2%) were from low income families. The overall prevalence of S. pneumoniae infection in the study was 18% (95% CI 14.4-22.2). No window (AOR=2.8 CI 1.1-7.6), no/non-exclusive breast-feeding (AOR= 2.1 CI 1.1-4.1), and previous URTI (AOR= 3.2 CI 1.7-6.1) were significantly associated with S. pneumoniae infection. The isolated organism showed drug resistance for Cotrimoxazole (35%), and Tetracycline (34%). The prevalence and antimicrobial resistance in this study were comparatively high. No window, non-exclusive breast-feeding and previous URTI were associated with S. pneumoniae infection. The isolated S. pneumoniae showed high drug resistance to cotrimoxazole and tetracycline.

Prevalence and antimicrobial resistance of Escherichia coli and Salmonella spp. in animal feed in Colombia.

To determine the prevalence and antimicrobial resistance of Escherichia coli and Salmonella spp. in animal feed samples collected between 2018 and 2021 in Colombia. This was a laboratory-based cross-sectional study using routine data from the program for inspection, surveillance, and control of animal feed at the Colombian Agriculture Institute. Samples of animal feed for swine, poultry, canine, feline, leporine, piscine, and equine species were processed for detection of E. coli and Salmonella spp. using enrichment and selective culture methods. Isolates were tested for antimicrobial susceptibility using an automated microdilution method. Of 1 748 animal feed samples analyzed, 83 (4.7%) were positive for E. coli and 66 (3.8%) for Salmonella spp. The presence of E. coli and Salmonella spp. was highest in feed for poultry (6.4% and 5.5%) and swine (6.1% and 4.3%). Antimicrobial resistance testing was performed in 27 (33%) E. coli isolates and 26 (39%) Salmonella isolates. Among E. coli, resistance was most frequently observed to ampicillin (44.5%) followed by cefazolin (33.3%), ciprofloxacin (29.6%), ampicillin/sulbactam (26%), and ceftriaxone (11.1%). The highest resistance levels in Salmonella spp. isolates were against cefazolin (7.7%) and piperacillin/tazobactam (7.7%). This is the first study from Colombia reporting on the prevalence and antimicrobial resistance of E. coli and Salmonella spp. in animal feed samples. Its results establish a baseline over a wide geographical distribution in Colombia. It highlights the need to integrate antimicrobial resistance surveillance in animal feed due to the emergence of resistant bacteria in this important stage of the supply chain.

Detection of genes encoding enterotoxins (SEA-D) in Staphylococcus aureus strains isolated from workers' nasal samples and creamy pastries of Shiraz confectioneries.

Staphylococcus aureus is responsible for the majority of food poisoning all around the world. Nasal carriers of S. aureus and foodstuffs need for handling are important sources and vehicles to transmit this pathogen to ready-to-eat foods. According to hygienic standards, confectioners should not be contaminated with S. aureus. This study aimed to detect nasal carriers and creamy pastries contaminated with enterotoxigenic S. aureus in confectioneries of Shiraz, Iran. From the confectioneries of Shiraz city, 27 places in the north, south, center, west, and east areas were selected randomly then 100 creamy pastries samples and 117 nasal swabs were collected. Bacteriological and biochemical tests were performed to isolate S. aureus. The polymerase chain reaction (PCR) test was used to identify the virulence and enterotoxins genes of the S. aureus isolates. Agar disk diffusion was performed to find out the antibiotic resistance of the isolates. Results revealed that 16.24 and 33 percent of workers and creamy pastries were contaminated with S. aureus respectively. Also, 100%, 37%, 58%, and 6% of nasal samples harbored femA, mecA, sea, and sec genes respectively. According to the results 97%, 70%, 54.5%, and 6% of creamy pastries isolates harbored femA, mecA, sea, and sec genes respectively. No isolate carried seb and sed genes. The results also showed that 41.5% of nasals and 5.5% of creamy pastry isolates harbored both sea and sec genes. The sea was the most common enterotoxin gene observed in nasal and creamy pastries. The results of the antimicrobial resistance test showed that 68.42% of nasal and 48.48% of creamy pastry isolates were resistant to cefocxitn (FOX) respectively. Both nasal (89%) and creamy pastry (82%) isolates presented the highest resistance to penicillin (P) and the most sensitivity to trimethoprim-sulphamethoxazole (SXT) (94%). Most of the isolates were sensitive to erythromycin (E), aztreonam (AZM), tetracycline (TE), trimethoprim (TMP), and ciprofloxacin (CP). Isolates of S. aureus harboring multi-enterotoxin genes were resistant to more antibiotics than others. The presence of enterotoxigenic S. aureus in the workers' nasal samples and creamy pastries of Shiraz confectioneries was high which is a potential public health hazard.

Microbial Contamination and Antibiotic Resistance in Marketed Food in Bangladesh: Current Situation and Possible Improvements.

Antimicrobial resistance (AMR) is a public health problem worldwide. Bangladesh, like its neighboring countries, faces many public health challenges, including access to safe food, inadequate food surveillance, as well as increasing AMR. This study investigated bacterial contamination and the AMR profile of pathogens in marketed food in Bangladesh and explored barriers to reducing AMR in the country. We collected 366 tomatoes, 359 chicken and 249 fish samples from 732 vendors in traditional markets in urban, peri-urban and rural areas in Bangladesh, as well as from 121 modern retails in Dhaka capital to analyse Vibrio cholerae and Escherichia coli in fish, Salmonella in chicken, and Salmonella and E. coli in tomatoes. Antibiotic susceptibility against 11 antibiotics was tested using a disc diffusion test and interpreted by an automated zone inhibition reader. In addition, a qualitative study using key informant interviews was conducted to explore antimicrobial use and AMR reduction potential in Bangladesh. We found E. coli in 14.21% of tomatoes and 26.91% of fish samples, while 7.38% of tomatoes and 17.27% of chicken were positive for Salmonella, and 44.98% of fish were positive for Vibrio cholerae. In total 231/319 (72.4%) of all pathogens isolated were multidrug-resistant (MDR) (resistant to three or more antibiotic groups). Qualitative interviews revealed an inadequate surveillance system for antibiotic use and AMR in Bangladesh, especially in the agriculture sector. To be able to fully understand the human health risks from bacterial hazards in the food and the AMR situation in Bangladesh, a nationwide study with a one health approach should be conducted, within all sectors, including AMR testing as well as assessment of the antimicrobial use and its drivers.

Genotyping and antimicrobial resistance profiles of chicken originated Salmonella Enteritidis isolates.

Salmonellosis is a common foodborne zoonosis worldwide. The most common Salmonella serovar in humans is Salmonella enterica subsp. enterica serovar Enteritidis (50.3%) in the world. The main transmission route for S. Enteritidis is consumption of contaminated poultry products. Therefore, it is important to determine the diversity and spread of chicken-originated S. Enteritidis isolates in order to monitor and control salmonellosis. Pulsed-field gel electrophoresis (PFGE) and multiple locus variable number of tandem repeats analysis (MLVA) are frequently used for typing of S. Enteritidis isolates. This study aimed to determine the antimicrobial resistance (AMR) profiles and MLVA and PFGE genotypes of chicken-originated S. Enteritidis isolates. A total of 200 S. Enteritidis isolated from chicken broiler, layer, and breeder flocks from different locations in Turkey were investigated by Kirby-Bauer disk diffusion method, PFGE, and MLVA. The AMR test indicated that 57% of the S. Enteritidis isolates were susceptible to all antimicrobials, while 39% were resistant to at least one antimicrobial. The highest resistance (25%) was against ampicillin. Multi-drug resistance rate was low (21%) and mostly from broiler flocks (93%). All isolates were genotyped into 32 different PFGE genotypes (PT) and 34 different MLVA genotypes (MT). The dominant genotypes were PT6 (12.5%) and MT22 (50%). In specific sample groups, there was a correlation between genotypes, breeding type, geographic location, and isolation years of the isolates. There was no significant difference in the discrimination power of PFGE and MLVA. However, MLVA was more suitable for large sample groups and routine genotyping because it was easier, quicker, and less labor-intensive to use.

Phenotypic and genotypic characterization of antimicrobial resistance in commonly isolated Salmonella serovars from chickens

Salmonellosis caused by Salmonella agents is the second most common zoonotic infection in humans. In recent years, Salmonella's increasing antimicrobial resistance (AMR) has been a concern. The major transmission route of Salmonella is consumption of contaminated poultry products. Therefore, monitoring of antimicrobial resistance in chicken-originated Salmonella is critically important. This study investigated AMR in four commonly isolated Salmonella serovars from chickens, namely Salmonella Enteritidis (S. Enteritidis), Salmonella Infantis (S. Infantis), Salmonella Kentucky (S. Kentucky), and Salmonella Typhimurium (S. Typhimurium). A total of 133 isolates were examined by phenotypic and genotypic AMR characterization. Resistance to 14 different antimicrobials and eight resistance genes were investigated in all isolates. The AMR test indicated that there was no resistant isolate to all antimicrobials while 14.3% were susceptible to all antimicrobials. The highest resistance was to sulfonamides (57.1%), nalidixic acid (48.1%), and tetracycline (39.1%). The highest susceptibilities were to cefotaxime (86.5%), cefoxitin (92.5%), ceftazidime (78.2%), and ceftriaxone (97%). S. Infantis and S. Kentucky isolates had higher resistance to all antimicrobials than S. Enteritidis and S. Typhimurium isolates. Significantly high multidrug-resistance (MDR) was detected in 50.4% of all isolates, although MDR prevalence varied widely between serovars: 78.7% of all S. Infantis isolates were MDR whereas only 18.8% of S. Enteritidis isolates were MDR. The most prevalent resistance genes were tetA (35.2%) and sul1 (31.6%), with 12.5% and 3.1% of S. Enteritidis isolates being positive for tetA and sul1, respectively, whereas 17.4% and 8.7% of S. Typhimurium isolates were positive. These rather low prevalence rates are probably due to effective monitoring of these serovars by control programs in Türkiye. The nondetection of mcr1 and mcr2 can be explained by the rare use of colistin in chicken flocks in Türkiye. The obtained findings emphasize the importance of AMR monitoring for Salmonella and the risks of chicken-originated isolates to humans.

Reporting of Antimicrobial Resistance from Blood Cultures, an Antibacterial Resistance Leadership Group Survey Summary: Resistance Marker Reporting Practices from Positive Blood Cultures.

We assessed how laboratories use and handle reporting of results of rapid diagnostics performed on positive blood culture broths, with a focus on antimicrobial resistance (AMR) markers. A survey assembled by the Antibacterial Resistance Leadership Group Diagnostics Committee was circulated from December 2020 to May 2021. The survey was sent to local hospitals, shared on the ClinMicroNet and Division C listservs, and included in a College of American Pathologists proficiency testing survey. Ninety-six laboratories of various sizes across the United States (95%) and outside of the United States (5%) participated. Of the laboratories that had at least 1 rapid diagnostic in place (94%), significant heterogeneity in methods used and reporting practices was found across community (52%) and academic (40%) laboratories serving hospitals of various sizes. Respondents had implemented 1 to 6 different panels/platforms for a total of 31 permutations. Methods of reporting rapid organism identification and AMR results varied from listing all targets as "detected"/"not detected" (16-22%) without interpretive guidance, to interpreting results (23-42%), or providing therapeutic guidance comments to patient-facing healthcare teams (3-17%). Current approaches to reporting molecular AMR test results from positive blood culture vary significantly across clinical laboratories. Providing interpretative comments with therapeutic guidance alongside results reported may assist clinicians who are not well-versed in genetic mechanisms of AMR. However, this is currently not being done in all clinical laboratories. Standardized strategies for AMR gene result reporting are needed.

Characterization of coagulase-positive Staphylococcus spp., antimicrobial resistance profile, and presence of enterotoxin-producing genes in goat milk in Paraná State

Goat milk presents interesting characteristics to consumers, but the production of this food faces several challenges that influence its quality. Process failures from milking to processing and commercialization can expose milk to contamination by pathogenic microorganisms, including the coagulase-positive Staphylococcus group. Although Staphylococcus aureus is the most relevant species in mastitis and food poisoning, other species in this group are also important, especially those related to food poisoning. This study aimed to identify the coagulase-positive Staphylococcus species in goat milk using biochemical tests, determine the prevalence of antimicrobial resistance using the disc diffusion test, and investigate enterotoxin-producing genes, sea, seb, sec, sed, see, seg, seh, and sei by multiplex PCR. A total of 384 coagulase-positive Staphylococcus strains obtained from raw goat milk collected from nine farms in Paraná during four seasons of the year were studied. Biochemical tests showed that 85.69% of the 384 strains were S. aureus, followed by 9.38% of S. intermedius, 4.17% of S. hyicus and 0.78% of S. delphini. For the antimicrobial resistance test, up to three strains of each species identified as coagulase-positive Staphylococci were chosen from each farm, with a total of 74 strains. Of these, 27% (20/74) showed resistance to at least one antibiotic, and among all penicillin-resistant strains, 45% (9/20) also presented resistance to tetracycline. In the search for staphylococcal enterotoxin-producing genes, 49 S. aureus strains were studied; among them, 40.81% (20/49) presented enterotoxin-producing genes. The highest prevalence was detected for the sec gene, which was present in 22.44% (11/49) of the strains, followed by the seh gene in 18.36% (9/49), and the see gene was detected in 4.08% (2/49). It was concluded that S. aureus was the predominant species in raw goat milk, with a high prevalence of penicillin- and tetracycline-resistant Staphylococci and a significant number of strains with staphylococcal enterotoxin-producing genes. The strains studied carried enterotoxin-producing genes involved in food poisoning in humans, indicating that goat milk is a possible source of contamination and, therefore, a potential danger to public health.

High rate of fluoroquinolone resistant Neisseria gonorrhoeae detected by molecular surveillance of antimicrobial resistance determinants in Bulgaria

Antimicrobial-resistant Neisseria gonorrhoeae is a major public health concern. The surveillance of antimicrobial resistance benefits from rapid and accurate molecular techniques in molecular diagnostics to facilitate individualised medicine and antimicrobial stewardship. To support the recommendations for empirical treatment of gonococcal infections in Bulgaria, we investigated N. gonorrhoeae-positive clinical specimens from 2018 to 2021 for the presence of genetic determinants associated with antimicrobial resistance. N. gonorrhoeae-positive samples stored at the National Center of Infectious and Parasitic Diseases during the four-year study period were retrospectively analysed by polymerase chain reaction and DNA sequencing assays for resistance determinants to fluoroquinolones, third-generation cephalosporins and macrolides. The detected determinants indicated a high rate of fluoroquinolone resistance (59%), very low level of decreased susceptibility to third-generation cephalosporins (3%) but no macrolide resistance (0%). These findings validate the utilisation of the international guidelines’ recommendations for empirical dual therapy with ceftriaxone/cefixime and azithromycin in Bulgaria. Because of the high fluoroquinolone resistance rate, ciprofloxacin should only be considered as treatment if phenotypic or molecular antimicrobial susceptibility data indicate susceptibility to ciprofloxacin. For the purposes of surveillance and individualised medicine, molecular assays for resistance determinants could complement culture-based phenotypic gonococcal antimicrobial resistance testing.

Virulence Profiles and Antimicrobial Resistance of Streptococcus agalactiae Infective and Colonizing Strains from Argentina.

Streptococcus agalactiae (GBS) is a colonizing agent in pregnant women, the main cause of invasive neonatal infections, and the reason of serious diseases in non-pregnant adults. Several virulence determinants are involved in the pathogenesis. These include capsular polysaccharide, surface-localized proteins, and toxins. Penicillin is considered the first choice antibiotic for the treatment and prophylaxis; erythromycin, clindamycin and fluoroquinolones are recommended alternatives for penicillin-allergic GBS carriers or patients. Our objective was to investigate the virulence genetic characteristics and the antimicrobial susceptibility of 162 GBS colonizing and infective isolates recovered in Argentina. Serotypes Ia and III were the most prevalent ones, followed by Ib, II, V, IV and non-typeable. In relation to the 13 virulence genes screened, cpsA, cylE, hylB, lmb, and scpB were the most prevalent and could be postulated as vaccine epitopes; bca, rib, bac, hvgA, spb1, PI, PI-2a, and PI-2b were detected in lesser frequencies. No significant association was found between serotypes or virulence genes and colonizing or infective isolates but, on the contrary, significant association was observed between some genes and the most prevalent serotypes, la and III. The cluster analysis showed 52 virulence profiles and, antimicrobial resistance tests, 16 profiles, some with up to 4 resistances. Tetracycline resistance was significantly associated with colonizing isolates. Genes tetM and ermB conferring resistance to tetracyclines and macrolides, respectively, were the most commonly identified. Our findings show that GBS colonizing and infective isolates circulating in Argentina share similar features in terms of serotype and virulence genes and show a high level of antimicrobial resistance.

Molecular Investigation of Some Beta-lactamase Genes by PCR and DNA Sequencing Techniques in clinical Escherichia coli

In this study, out of 50 isolates of some nosocomial infections from some Baghdad hospitals, only 13 (26%) were identified as Escherichia coli. Depending on selective media, morphological and biochemical tests the species was then confirmed by molecular methods. Later on antimicrobial resistance test was performed by the Kirby-Bauer method. The molecular characterization of blaTEM and blaCTX-M genes in different clinical isolates of E. coli was done through polymerase chain reaction (PCR) by utilizing special primers. These genes were positive to only 4 (30.7%) isolates. The sequence of nucleotides of positive genes was carried out for four isolates. The results showed that there was no variance in the nucleotide sequence between Iraqi isolates compared with the global isolates, and that they were 100% identical to many genera of Enterobacteriaceae. Finally, due to the indiscriminate use of antibiotics, these genes in human strains were likely the source of widespread drug resistance.

Species diversity and antimicrobial resistance patterns of Enterococcus spp. isolated from mastitis cases, milking machine and the environment of dairy cows.

The objectives were to study the diversity of Enterococcus spp. isolated from mastitis cases, milking equipment and the environment of dairy cows; and to determine invitro resistance of isolates to antimicrobials that are relevant to human and animal health. Ten dairy farms were visited to collect samples from mastitis cases, faeces, bedding, aisles, water and milking equipment. Identification of Enterococcus at the species level and antimicrobial resistance testing was performed by MALDI-TOF and the disk-diffusion method, respectively. Of 365 isolates, Enterococcus hirae was the most prevalent, being more likely to be isolated from faeces than from milk (odds ratio (OR) = 39·2), liners (OR = 5·4) or bedding (OR = 2·2). Enterococcus saccharolyticus was the most prevalent in milk samples. The chances of isolating Enterococcus faecalis from milk were higher than from aisles (OR = 12·5), faeces (OR = 5·3), bedding (OR = 3·6) or liners (OR = 3·0). The odds of isolating Enterococcus faecium from faeces were higher than from liners (OR = 7·3), bedding (OR = 2·5) or aisles (OR = 2·4). Of 360 tested isolates, 1·9, 0·3 and 0·6% were resistant to penicillin, vancomycin and teicoplanin, respectively. Our results suggest that Enterococcus species can occupy specific ecological niches on dairy farms and pose a risk to public and animal health.

Performance of a Machine Learning-Based Methicillin Resistance of Staphylococcus aureus Identification System Using MALDI-TOF MS and Comparison of the Accuracy according to SCCmec Types.

The prompt presumptive identification of methicillin-resistant Staphylococcus aureus (MRSA) using matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS) can aid in early clinical management and infection control during routine bacterial identification procedures. This study applied a machine learning approach to MALDI-TOF peaks for the presumptive identification of MRSA and compared the accuracy according to staphylococcal cassette chromosome mec (SCCmec) types. We analyzed 194 S. aureus clinical isolates to evaluate the machine learning-based identification system (AMRQuest software, v.2.1, ASTA: Suwon, Korea), which was constructed with 359 S. aureus clinical isolates for the learning dataset. This system showed a sensitivity of 91.8%, specificity of 83.3%, and accuracy of 87.6% in distinguishing MRSA. For SCCmec II and IVA types, common MRSA types in a hospital context, the accuracy was 95.4% and 96.1%, respectively, while for the SCCmec IV type, it was 21.4%. The accuracy was 90.9% for methicillin-susceptible S. aureus. This presumptive MRSA identification system may be helpful for the management of patients before the performance of routine antimicrobial resistance testing. Further optimization of the machine learning model with more datasets could help achieve rapid identification of MRSA with less effort in routine clinical procedures using MALDI-TOF MS as an identification method.

A Simple and Rapid Low-Cost Procedure for Detection of Vancomycin-Resistance Genes in Enterococci Reveals an Outbreak of Vancomycin-Variable Enterococcus faecium.

The detection of resistance to vancomycin in enterococci cultured from patients is important for the treatment of individual patients and for the prevention of hospital transmission. Phenotypic antimicrobial resistance tests may fail to detect potential vancomycin-resistant enterococci. We have developed and tested a PCR based procedure for routine screening for vancomycin-resistance genes in clinical samples with enterococci. Primary cultures from diagnostic samples reported with growth of Enterococcus faecium or E. facalis were tested for vanA and vanB genes by real-time PCR without the isolation of specific bacteria. Up to ten samples were pooled and tested in each real-time PCR reaction, with subsequent individual testing of cultures from positive pools. In a one-month test period in 2017 vanA gene was detected in one out of 340 urine samples with vancomycin-susceptible enterococci reported from diagnostic culture. A second test period in 2018 included 357 urine samples, and vanA gene was detected in samples from eight patients. Subsequently, all urine samples reported with growth of E. faecium during a period of one year were tested. Fifty-eight individuals were identified with enterococci, carrying the vanA gene not previously detected. Routine molecular testing of primary culture material from patient samples may improve the detection of hospitalized patients carrying E. faecium with resistance genes to vancomycin.

Characterization of the virulence, agr typing and antimicrobial resistance profile of Staphylococcus aureus strains isolated from food handlers in Brazil.

Staphylococcus aureus is one of the main pathogens associated with foodborne outbreaks in Brazil and food handlers can carry toxigenic and resistant S. aureus strains. The aims of this study were to verify the frequency of virulence genes, to identify the agr groups and to determine the antimicrobial resistance profile of S. aureus strains isolated from food handlers of pilot kitchens located in São Paulo, Brazil. A total of 74 strains of the Staphylococcus genus were detected and 50% were identified as of the species S. aureus. The enterotoxin genes detection, tst and luk-PV detection, agr typing, mecA detection, ccr complex detection and SCCmec typing were performed using PCR. The antimicrobial resistance testing was performed by the disk diffusion method. The enterotoxin genes were identified in 36 S. aureus, including sea (83.8%). The tst gene was detected in 18.92% of the strains and the luk-PV was detected in only one isolate. Agr typing classified 58.3% of the strains as type I. Seven (18.92%) strains were classified as MRSA and the ccr2 complex was detected in six of these isolates. The SCCmec typing characterized strains as type II, III, IV and V. Moreover, there were also a greater number of resistant strains to penicillin (83.78%) and clarithromycin (67.57%). In conclusion, the study revealed a significant prevalence of S. aureus, and the presence of different virulence genes and a worrying resistance profile in S. aureus strains isolated from food handlers in this country.

Susceptibility Antibiotics of Bacteria Causing Urinary Tract Infection in Pregnant Women Infected with COVID-19

Urinary tract infections (UTIs) are a significant issue in women of all ages, but they are especially common during pregnancy. Co-infection of the Coronavirus Disease 2019 (COVID-19) with UTIs in pregnant females is a hot topic today, as it may be linked to various conditions. Furthermore, understanding the types of bacteria that cause UTIs and related antimicrobial resistance may aid the clinician in selecting the appropriate empirical treatment. This study aimed to isolate and characterize causative agents of UTIs and determine antimicrobial treatment sensitivity patterns among pregnant women diagnosed with the COVID-19 admitted to Teaching Hospital X in Iraq. Antimicrobial drug resistance testing was performed on 42 mid-stream urine samples that had been prepared for culture. Escherichia coli (18/42.85%), Klebsiella pneumoniae (9/21.45%), Streptococcus galactica (7/16.66%), and Proteus mirabilis (4/9.52%) were the bacteria isolated and diagnosed from pregnant women. The infections Pseudomonas aeruginosa and Staphylococcus aureus were the least common (2/4.7). In conclusion, the isolated uro-pathogens showed high resistance to Gentamicin, Cefuroxime, Ceftriaxone and sensitive to Ampicillin and Nitrofurantoin. The common cause of UTIs among pregnant women infected with the COVID-19 was discovered to be E. coli. Before recommending therapy, culture and sensitivity testing of isolates from urine tests should be conducted on a regular basis.

Genomic epidemiological analysis of Klebsiella pneumoniae from Portuguese hospitals reveals insights into circulating antimicrobial resistance

Klebsiella pneumoniae (Kp) bacteria are an increasing threat to public health and represent one of the most concerning pathogens involved in life-threatening infections and antimicrobial resistance (AMR). To understand the epidemiology of AMR of Kp in Portugal, we analysed whole genome sequencing, susceptibility testing and other meta data on 509 isolates collected nationwide from 16 hospitals and environmental settings between years 1980 and 2019. Predominant sequence types (STs) included ST15 (n = 161, 32%), ST147 (n = 36, 7%), ST14 (n = 26, 5%) or ST13 (n = 26, 5%), while 31% of isolates belonged to STs with fewer than 10 isolates. AMR testing revealed widespread resistance to aminoglycosides, fluoroquinolones, cephalosporins and carbapenems. The most common carbapenemase gene was blaKPC-3. Whilst the distribution of AMR linked plasmids appears uncorrelated with ST, their frequency has changed over time. Before year 2010, the dominant plasmid group was associated with the extended spectrum beta-lactamase gene blaCTX-M-15, but this group appears to have been displaced by another carrying the blaKPC-3 gene. Co-carriage of blaCTX-M and blaKPC-3 was uncommon. Our results from the largest genomics study of Kp in Portugal highlight the active transmission of strains with AMR genes and provide a baseline set of variants for future resistance monitoring and epidemiological studies.

Repurposing Drugs to Combat Drug Resistance in Leprosy: A Review of Opportunities.

Leprosy is caused by extremely slow-growing and uncultivated mycobacterial pathogens, namely Mycobacterium leprae and M. lepromatosis. Nearly 95% of the new cases of leprosy recorded globally are found in India, Brazil, and 20 other priority countries (WHO, 2019), of which nearly two-third of the cases are reported in India alone. Currently, leprosy is treated with dapsone, rifampicin, and clofazimine, also known as multi-drug therapy (MDT), as per the recommendations of WHO since 1981. Still, the number of new leprosy cases recorded globally has remained constant in last one-decade, and resistance to multiple drugs has been documented in various parts of the world, even though relapses are rare in patients treated with MDT. Antimicrobial resistance testing against M. leprae or the evaluation of the anti-leprosy activity of new drugs remains a challenge as leprosy bacilli cannot grow in vitro. Besides, developing a new drug against leprosy through conventional drug development process is not economically attractive or viable for pharma companies. Therefore, a promising alternative is the repurposing of existing drugs/approved medications or their derivatives for assessing their anti-leprosy potential. It is an efficient method to identify novel medicinal and therapeutic properties of approved drug molecules. Any combinatorial chemotherapy that combines these repurposed drugs with the existing first-line (MDT) and second-line drugs could improve the bactericidal and synergistic effects against these notorious bacteria and can help in achieving the much-cherished goal of "leprosy-free world". This review highlights novel opportunities for drug repurposing to combat resistance to current therapeutic approaches.