Penicillin-resistant Isolates Research Articles

Gene Expression Of Some Antibiotic Resistance Mechanisms In Neisseria Gonorrhoeae Isolated From Iraqi Patients

Background: Treatment of N. gonorrhoeae infection is restricted by the rising commonness of multidrug-resistance strains. The point of this study was to evaluate antibiotic resistance and mRNA expression of penA, pilQ, norM, farA, and mtrE of N. gonorrhea. Method: one hindered samples collected from patients infected with N. gonorrhoeae. The bacteria were isolated and diagnosed using culture media and biochemical tests. Using the MIC method, the sensitivity of bacteria to penicillin, ciprofloxacin, ceftriaxone, spectinomycin, tetracycline, and azithromycin was determined. After RNA extraction and cDNA production, gene expression of penA, pilQ, norM, farA, and mtrE was assessed using real time -qPCR. Results: we found all isolates sensitive to ceftriaxone and azithromycin. Clear increase in penA, pilQ, farA, and mtrE gene expression of penicillin-resistant isolates (1.571, 3.135, 0.010 and 34.24 respectively), while norM gene expression decreased in those isolates. High gene expression for pilQ, farA, and mtrE among isolates resistant to ciprofloxacin (0.0041, 0.00072, 0.011 respectively). Tetracycline-resistant isolates appeared to have higher gene expression for penA, norM, farA, and mtrE (0.0033, 8.669, 37.85, 15.84 respectively). Spectinomycin-resistant isolates have high gene expression for penA, pilQ, norM, farA, and mtrE (0.222, 0.021, 0.036, 0.0002, 19.63 respectively) . Moreover, gene expression of penA, norM, and farA increased in males (6.938, 1.095, 7.946 respectively) while the rate of gene expression for pilQ and mtrE increased in females (0.161 and 7.832 respectively). Conclusion: gene expression of penA, pilQ, norM, farA, and mtrE increased in most antibiotics resistance isolates especially of bacterial infection that isolated from males.

2162. In Vitro Antimicrobial Activity of Ceftobiprole against Streptococcus pneumoniae Isolates from the United States (2016–2020)

Abstract Background Ceftobiprole (BPR) is an advanced-generation cephalosporin approved in Europe and many non-European countries for the treatment of community-acquired pneumonia and non–ventilator-associated hospital-acquired pneumonia in adults caused by indicated species, including Streptococcus pneumoniae (SPN). This study evaluated the in vitro antimicrobial activity of BPR and comparator agents against recent SPN isolates causing lower respiratory tract infections in patients at medical centers in the United States (US). Methods A total of 2,793 SPN isolates from 32 US medical centers (2016–2020) were collected from patients with lower respiratory tract infections. Isolates were tested for antimicrobial susceptibility using the Clinical and Laboratory Standards Institute (CLSI) broth microdilution method. MIC interpretations for BPR and comparators utilized European Committee on Antimicrobial Susceptibility Testing (EUCAST) or CLSI criteria, respectively. The EUCAST BPR susceptibility breakpoint for SPN was 0.5 mg/L. Results BPR inhibited 99.5% of all SPN isolates at the EUCAST breakpoint (MIC50/90, 0.015/0.25 mg/L). The MIC50/90 values were stable over all 5 surveillance years. The following comparator agents displayed lower activity against the isolate set: clindamycin (85.5% susceptible [S]), erythromycin (53.2% S), penicillin (63.2% S using oral breakpoints), tetracycline (79.4% S), and trimethoprim-sulfamethoxazole (72.8% S). BPR maintained excellent in vitro activity against each of the isolate subsets resistant to these comparator antimicrobials (95.4–98.8% S to BPR depending on the comparator antimicrobial; 95.4% of the penicillin-resistant isolates were S to BPR). This SPN isolate set was also 99.4% S to levofloxacin, 97.4% S to ceftriaxone (using non-meningitis breakpoints), and 100% S to linezolid and vancomycin. Conclusion BPR exhibited potent in vitro antimicrobial activity against recent SPN clinical isolates collected in US medical centers (2016–2020). These data indicate that BPR merits further investigation as a potential option for the treatment of lower respiratory tract infections caused by SPN in the US. Disclosures Leonard R. Duncan, PhD, AbbVie: Grant/Research Support|Basilea: Grant/Research Support|CorMedix: Grant/Research Support|Melinta: Grant/Research Support|Pfizer: Grant/Research Support Mariana Castanheira, PhD, AbbVie: Grant/Research Support|Basilea: Grant/Research Support|bioMerieux: Grant/Research Support|Cipla: Grant/Research Support|CorMedix: Grant/Research Support|Entasis: Grant/Research Support|Melinta: Grant/Research Support|Paratek: Grant/Research Support|Pfizer: Grant/Research Support|Shionogi: Grant/Research Support Jennifer Smart, PhD, Basilea Pharmaceutica International Ltd, Allschwil, Switzerland: Stocks/Bonds Mark E. Jones, PhD, Astellas Pharma Global Development, Inc: Support for the present publication|Basilea Pharmaceutica International Ltd: Employee of Basilea Pharmaceutica International Ltd|Basilea Pharmaceutica International Ltd: Stocks/Bonds Rodrigo E. Mendes, PhD, AbbVie: Grant/Research Support|Basilea: Grant/Research Support|Cipla: Grant/Research Support|Entasis: Grant/Research Support|GSK: Grant/Research Support|Paratek: Grant/Research Support|Pfizer: Grant/Research Support|Shionogi: Grant/Research Support

In vitro activity of ceftaroline against bacterial isolates causing skin and soft tissue and respiratory tract infections collected in Latin American countries, ATLAS program 2016–2020

Objectives: Ceftaroline, a broad-spectrum cephalosporin has activity against Gram-positive and several Gram-negative bacteria (GNB). This study aimed to evaluate the antimicrobial activity of ceftaroline and comparators against isolates causing skin and soft tissue infections (SSTIs) and respiratory tract infections (RTIs) collected in Latin America (LATAM) in 2016-2020 as part of the Antimicrobial Testing Leadership and Surveillance program (ATLAS). Methods: Minimum inhibitory concentrations were determined using both Clinical and Laboratory Standards Institute (CLSI) and European Committee on Antimicrobial Susceptibility Testing (EUCAST) criteria. Results: Ceftaroline demonstrated potent activity against methicillin-susceptible Staphylococcus aureus (CLSI/EUCAST: MIC90 0.25 mg/L; susceptibility 100%), whereas activity against methicillin-resistant S. aureus varied for SSTIs (MIC90 1 mg/L; susceptibility 92.5%) and RTIs isolates (MIC90 2 mg/L; susceptibility 72.9%) isolates. For Streptococcus pneumoniae, particularly penicillin-resistant isolates commonly causing respiratory infections, high ceftaroline activity (MIC90 0.25 mg/L; susceptibility 100%/98.4%) was noted. All isolates of β-hemolytic streptococci were susceptible to ceftaroline [S. agalactiae: MIC90 0.03 mg/L (SSTIs); MIC90 0.015 mg/L (RTIs); susceptibility 100%; S. pyogenes: MIC90 0.008 mg/L; susceptibility 100%). Ceftaroline was highly active against Haemophilus influenzae including β-lactamase positive isolates (MIC90 0.06 mg/L; susceptibility 100%/85.7%). Ceftaroline demonstrated high activity against non-ESBL-producing GNB (E. coli: MIC90 0.5 mg/L, susceptibility 91.9%; K. pneumoniae: MIC90 0.25 mg/L, susceptibility 95.1%; K. oxytoca, MIC90 0.5 mg/L; susceptibility 95.7%). Conclusion: Ceftaroline was active against the recent collection of bacterial pathogens commonly causing SSTIs and RTIs in LATAM. Local and regional surveillance of antimicrobial resistance patterns are crucial to understand evolving resistance and guide treatment management.

Emergence of ciprofloxacin- and penicillin-resistant Neisseria meningitidis isolates in Japan between 2003 and 2020 and its genetic features.

Although we previously reported that some meningococcal isolates in Japan were resistant to penicillin (PCG) and ciprofloxacin (CIP), the antibiotic susceptibilities of Neisseria meningitidis isolates obtained in Japan remained unclear. In the present study, 290 N. meningitidis isolates in Japan between 2003 and 2020 were examined for the sensitivities to eight antibiotics (azithromycin, ceftriaxone, ciprofloxacin, chloramphenicol, meropenem, minocycline, penicillin, and rifampicin). All isolates were susceptible to chloramphenicol, ceftriaxone, meropenem, minocycline, and rifampicin while two were resistant to azithromycin. Penicillin- and ciprofloxacin-resistant and -intermediate isolates (PCGR, CIPR, PCGI and CIPI, respectively) were also identified. Based on our previous findings from whole genome sequence analysis, approximately 40% of PCGI were associated with ST-11026 and cc2057 meningococci, both of which were unique to Japan. Moreover, the majority of ST-11026 meningococci were CIPR or CIPI. Sensitivities to PCG and CIP were closely associated with genetic features, which indicated that, at least for Japanese meningococcal isolates, PCGR/I or CIPI/R would be less likely to be horizontally conferred from other neisserial genomes by transferring of the genes responsible (penA and gyrA genes, respectively), but rather that ancestral N. meningitidis strains conferring PCGR/I or CIPI/R phenotypes clonally disseminated in Japan.

Impact of the progressive uptake of pneumococcal conjugate vaccines on the epidemiology and antimicrobial resistance of invasive pneumococcal disease in Gipuzkoa, northern Spain, 1998-2022.

To analyze the impact of pneumococcal conjugate vaccines (PCVs) on the incidence of invasive pneumococcal diseases (IPDs) and pneumococcal antibiotic resistance in Gipuzkoa, northern Spain for a 25 years period. All cases of IPD confirmed by culture between 1998 and 2022 in a population of around 427,416 people were included. Pneumococci were serotyped and antimicrobial susceptibility was assessed by the EUCAST guidelines. Overall, 1,516 S. pneumoniae isolates were collected. Annual IPD incidence rates (per 100,000 people) declined from 19.9 in 1998-2001 to 11.5 in 2017-19 (42.2% reduction), especially in vaccinated children (from 46.7 to 24.9) and non-vaccinated older adult individuals (from 48.0 to 23.6). After PCV13 introduction, the decrease in the incidence of infections caused by PCV13 serotypes was balanced by the increase in the incidence of non-PCV13 serotypes. In the pandemic year of 2020, IPD incidence was the lowest: 2.81. The annual incidence rates of penicillin-resistant isolates also decreased, from 4.91 in 1998-2001 to 1.49 in 2017-19 and 0.70 in 2020. Since 2017, serotypes 14, 19A, and 11A have been the most common penicillin-resistant types. The incidence of erythromycin-resistant strains declined, from 3.65 to 1.73 and 0.70 in the same years. PCV use was associated with declines in the incidence of IPD and the spread of non-vaccine serotypes, that balanced the beneficial effect off PCV13, some of them showing high rates of antibiotic resistance.

Antimicrobial susceptibility of staphylococci from bovine milk samples in routine microbiological mastitis analysis in Finland.

The most frequent reason for antimicrobial use in dairy herds is mastitis and knowledge about mastitis-causing pathogens and their antimicrobial susceptibility should guide treatment decisions. The overall objective of this study was to assess antimicrobial resistance (AMR) of staphylococci in mastitic milk samples in Finland. MALDI-ToF MS identified a total of 504 Staphylococcus isolates (260 S. aureus and 244 non-aureus staphylococci, NAS) originating from bovine mastitic milk samples. Phenotypic susceptibility against cefoxitin, ceftiofur, enrofloxacin, gentamycin, oxacillin, penicillin, and tetracycline was evaluated by disk diffusion method and the presence of blaZ, mecA, and mecC genes investigated by PCR. Nitrocefin test assessed these isolates' beta-lactamase production. The most common NAS species were S. simulans, S. epidermidis, S. chromogenes, and S. haemolyticus. In total, 26.6% of the isolates (18.5% of S. aureus and 35.2% of all NAS) carried the blaZ gene. Penicillin resistance, based on disk diffusion, was lower: 18.8% of all the isolates (9.3% of S. aureus and 28.9% of all NAS) were resistant. Based on the nitrocefin test, 21.5% of the isolates produced beta-lactamase (11.6% of S. aureus and 32.0% of all NAS). Between the Staphylococcus species, the proportion of penicillin-resistant isolates varied, being lowest in S. simulans and highest in S. epidermidis. Resistance to antimicrobials other than penicillin was rare. Of the eight NAS isolates carrying the mecA gene, six were S. epidermidis. One S. aureus isolate carried the mecC gene. Agreement beyond chance, assessed by kappa coefficient, between phenotypic and genotypic resistance tests, was moderate to substantial. Some phenotypically penicillin-susceptible staphylococci carried the blaZ gene but isolates without blaZ or mec genes rarely exhibited resistance, suggesting that the more reliable treatment choice may depend upon genotypic AMR testing. Our results support earlier findings that penicillin resistance is the only significant form of antimicrobial resistance among mastitis-causing staphylococci in Finland.

Resistance finding of Bacillus anthracis towards penicillin in East Java, Central Java, and Yogyakarta Provinces, Indonesia

Anthrax is a worldwide distributing zoonotic disease, caused by Bacillus anthracis, which occurs sporadically in Indonesia, particularly in the provinces of East Java, Central Java, and Yogyakarta, which are the working areas of the Disease Investigation Center (DIC) Wates. Penicillin has been the primary antimicrobial treatment recommended for anthrax since there has never been a report of resistance to this antibiotic in Indonesia. The objective of this research was to assess the sensitivity of B. anthracis isolates from Central Java, East Java, and Yogyakarta to penicillin and tetracycline. Sixteen B. anthracis isolates from DIC Wates collected between 1990-2021 recovered from environmental samples were used in this study. All isolates were identified by phenotype, then tested for sensitivity to penicillin and tetracycline by agar diffusion (Kirby-Bauer) and broth dilution method. The data obtained were compared with the standard and analyzed descriptively. The results showed that all isolates were B. anthracis. One of 16 isolates (6,25%) consistently showed resistance to penicillin, but was sensitive to tetracycline, while 15 isolates (93,75%) showed sensitive to both antibiotics. A penicillin-resistant isolate was soil sample from anthrax endemic area. In conclusion, there was B. anthracis isolate that was found resistance to penicillin. Therefore, tetracycline can be used as an alternative for anthrax treatment.

Australian Gonococcal Surveillance Programme Annual Report, 2022.

The Australian Gonococcal Surveillance Programme (AGSP) has continuously monitored antimicrobial resistance in Neisseria gonorrhoeae for more than 40 years. In 2022, a total of 8,199 isolates from patients in the public and private sectors, in all jurisdictions, were tested for in vitro antimicrobial susceptibility by standardisedmethods. The current treatment recommendation for gonorrhoea, for the majority of Australia, continues to be dual therapy with ceftriaxone and azithromycin. In 2022, of N. gonorrhoeae isolates tested, 0.51% (42/8,199) met the WHO criterion for ceftriaxone decreased susceptibility (DS), defined as a minimum inhibitory concentration value ≥ 0.125mg/L. Resistance to azithromycin was reported in 3.9% of N. gonorrhoeae isolates, proportionally stable since 2019. There were nine isolates with high-level resistance to azithromycin (MIC value ≥ 256 mg/L) reported in Australia: Queensland (4), New South Wales (3), Victoria (1) and non-remote Western Australia (1). This is the highest number detected annually by the AGSP. In 2022, penicillin resistance was found in 38.8% of gonococcal isolates, and ciprofloxacin resistance in 63.3%, however, there was considerable variation by jurisdiction. In some remote settings, penicillin resistance remains low; in these settings, penicillin continues to be recommended as part of an empiric therapy strategy. In 2022, in remote Northern Territory, one penicillin-resistant isolate was reported; in remote Western Australia, 11.8% of gonococcal isolates (9/76) were penicillin resistant. There were three ciprofloxacin-resistant isolates reported from remote Northern Territory; ciprofloxacin resistance rates remain comparatively low in remote Western Australia (6/76;7.9%).

Risk factors and pathogen characteristics associated with unfavorable outcomes among adults with pneumococcal meningitis in Japan, 2006 to 2016

PurposeIn this study, we aimed to clarify the risk factors associated with unfavorable outcomes in adults with pneumococcal meningitis (PnM). MethodsSurveillance was conducted between 2006 and 2016. Adults with PnM (n = 268) were followed up for outcomes within 28 days after admission using the Glasgow Outcome Scale (GOS). After classifying the patients into the unfavorable (GOS1–4) and favorable (GOS5) outcome groups, i) the underlying diseases, ii) biomarkers at admission, and iii) serotype, genotype, and antimicrobial susceptibility for all isolates were compared between both groups. ResultsOverall, 58.6% of patients with PnM survived,15.3% died, and 26.1% had sequelae. The number of living days in the GOS1 group was highly heterogeneous. Motor dysfunction, disturbance of consciousness, and hearing loss were the commonest sequelae. Of the underlying diseases identified in 68.9% of the PnM patients, liver and kidney diseases were significantly associated with unfavorable outcomes. Of the biomarkers, creatinine and blood urea nitrogen, followed by platelet and C-reactive protein had the most significant associations with unfavorable outcomes. There was a significant difference in the high protein concentrations in the cerebrospinal fluid between the groups. Serotypes 23F, 6C, 4, 23A, 22F, 10A, and 12F were associated with unfavorable outcomes. These serotypes were not penicillin-resistant isolates possessing three abnormal pbp genes (pbp1a, 2x, and 2b), except for 23F. The expected coverage rate of the pneumococcal conjugate vaccine (PCV) was 50.7% for PCV15 and 72.4% for PCV20. ConclusionsIn the introduction of PCV for adults, the risk factors for underlying diseases should be prioritized over age, and serotypes with unfavorable outcomes should be considered.

Phenotypic and Genotypic Characterization of Macrolide-Lincosamide-Streptogramin Resistance in Staphylococcus aureus Isolates from Bovine and Human

Abstract In this study, penicillin, oxacillin, and macrolide-lincosamide-streptogramin (MLS) resistance in S. aureus strains that were isolated from bovine mastitis cases, and human patients were investigated. Inducible clindamycin resistance (iML) was not found in 30 bovine isolates, while it was detected in 3 (10%) of 30 human isolates. MIC90 values of penicillin, oxacillin and macrolide-lincosamides (ML) were 2, 0.19, >256 µg/ml in bovine isolates and were 3, 3 and 0.19-1.5 µg/ml in human isolates, respectively. Streptogramin resistance was not found in both bovine and human isolates. Although the mecA gene was detected in all of the oxacillin resistant isolates, blaZ gene could not be detected in penicillin resistant isolates. The erm(B) gene was detected in 5 (38.6%) of 13 ML-resistant bovine isolates, and the mph(C) gene was detected in 2 (66.66%) of 3 human isolates. As a result, resistance to penicillin and oxacillin was found to be higher in human S. aureus isolates, while ML resistance was found to be higher in bovine isolates in this investigation. It was concluded that the presence of genes in extra-chromosomal elements associated to penicillin and macrolide resistance should be investigated. The data obtained from this study will contribute to the studies on antimicrobial susceptibility in the field of human and veterinary medicine.

Occurrence of virulence genes and methicillin-resistance in Staphylococcus aureus isolates causing subclinical bovine mastitis in Tiaret area, Algeria.

Mastitis remains the most frequent and the most expensive disease of dairy breeding. The objective of the study was to study S. aureus isolates collected from subclinical bovine mastitis in the Tiaret region, Algeria, by determining their antimicrobial susceptibility and their virulence traits. Sixty-two S. aureus isolates collected from subclinical bovine mastitis were studied by determining their antimicrobial susceptibility according to CLSI guidelines, and nine genes encoding virulence factors and resistance to methicillin and penicillin were determined by PCR. Multi-drug resistance was observed in 19 (30.64%) isolates and five (8%) were methicillin-resistant S. aureus (MRSA), four of them harbored the mecA gene; however, the mecC gene was not detected. Out of 59 penicillin-resistant isolates, 14 harbored the blaZ gene; one of them co-harbored the mecA gene. The following virulence genes were detected: eta (n=23; 37%), icaA (20; 32.2%), icaD (18; 29%), etb (16; 25.8%), luk E-D (14; 22.5%), and sea (6; 9.6%). The tsst-1, lukF/lukS, and luk-M genes were not detected. The occurrence of MRSA and multi-drug resistant (MDR) S. aureus isolates as well as genes encoding virulence factors playing an important role in the pathogenesis of subclinical bovine mastitis and of harmful potential to human is a cause for concern.

Going Back in Time: Increasing Penicillin Susceptibility among Methicillin-Susceptible Staphylococcus aureus Osteoarticular Infections in Children.

In the late 1940s to 1950s, Staphylococcus aureus isolates first-gained resistance to penicillin. Recently, some centers have described an increase in the proportion of methicillin susceptible S. aureus (MSSA) which are also susceptible to penicillin (PSSA). There are little data on the frequency of PSSA infections in children. We investigated the prevalence of penicillin susceptibility among pediatric MSSA acute hematogenous osteoarticular infection (OAI) isolates. MSSA OAI isolates were obtained through surveillance studies at Texas Children's and St. Louis Children's Hospitals from January 2011 to December 2019. All isolates underwent PCR for blaZ β-lactamase, PVL genes and agr group. All blaZ negative isolates then underwent penicillin MIC determination. blaZ negative isolates with penicillin MIC ≤ 0.125 μg/mL were considered PSSA. Multilocus sequence typing (MLST) was conducted on a subset of isolates. A total of 329 unique isolates were included in the study. The median patient age was 9.2 years (IQR:5.1 to 12.2). Overall, 6.7% of isolates were penicillin susceptible. No PSSA were detected prior to 2015 but increased yearly thereafter. By the final study year, 20.4% of isolates were PSSA (P = 0.001). PSSA were similar to penicillin-resistant MSSA (PR-MSSA) isolates in terms agr group and PVL carriage as well as clinical presentation and outcomes. PSSA were of distinct sequence types compared to PR-MSSA. PSSA appears to be increasing among OAI in U.S. children. Overall, PSSA isolates are associated with a similar clinical presentation as penicillin-resistant isolates. The potential for use of penicillin treatment in PSSA OAI warrants further study.

Australian Gonococcal Surveillance Programme Annual Report, 2021.

The Australian Gonococcal Surveillance Programme, established in 1981, has continuously monitored antimicrobial resistance in Neisseria gonorrhoeae for more than 40 years. In 2021, a total of 6,254 isolates from patients in the public and private sectors, in all jurisdictions, were tested for in vitro antimicrobial susceptibility by standardised methods. The current treatment recommendation for gonorrhoea, for the majority of Australia, continues to be dual therapy with ceftriaxone and azithromycin. In 2021, of isolates tested, 0.9% were reported nationally with decreased susceptibility (DS) to ceftriaxone (minimum inhibitory concentration [MIC] value ≥ 0.06 mg/L). There was one isolate from non-remote Western Australia that was resistant to ceftriaxone (MIC value ≥ 0.25 mg/L). Resistance to azithromycin (MIC value ≥ 1.0 mg/L) was reported nationally in 4.7% of N. gonorrhoeae isolates. This is increased from that reported in 2020 (3.9%) but similar to the percentage reported in 2019 (4.6%). Isolates with high-level resistance to azithromycin (MIC value ≥ 256 mg/L) are identified sporadically in Australia; none were reported in 2021. In 2021, penicillin resistance was found in 38% of gonococcal isolates nationally, and ciprofloxacin resistance in 53%; however, there is considerable variation by jurisdiction. In some remote settings, penicillin resistance remains low; in these settings, penicillin continues to be recommended as part of an empiric therapy strategy. In 2021, in remote Northern Territory, one penicillin-resistant isolate was reported, and in remote Western Australia 2/83 of gonococcal isolates (2.4 %) were penicillin resistant. There were two ciprofloxacin-resistant isolates reported from remote Northern Territory; ciprofloxacin resistance rates remain comparatively low in remote Western Australia (3/83; 3.6 %).

Whole-genome sequence analysis of high-level penicillin-resistant strains and antimicrobial susceptibility of Neisseria gonorrhoeae clinical isolates from Thailand.

The increasing rate of antimicrobial-resistant Neisseria gonorrhoeae poses a considerable public health threat due to the difficulty in treating gonococcal infections. This study examined antimicrobial resistance (AMR) to drugs recommended for gonorrhea treatment between 2015 and 2017, and the AMR determinants and genetic compositions of plasmids in 3 gonococcal strains with high-level penicillin resistance. We collected 117 N. gonorrhoeae isolates from patients with gonococcal infections who attended Siriraj Hospital, Bangkok, Thailand, between 2015 and 2017. Minimum inhibitory concentrations (MICs) of penicillin, tetracycline, ciprofloxacin, azithromycin, spectinomycin, cefixime, and ceftriaxone were determined by the agar dilution method. PCR amplification and sequencing of 23S rRNA and mtrR (a negative regulator of MtrCDE efflux pump) were performed. Whole genomes of 3 PPNG strains with high-level penicillin resistance (MIC ≥ 128 μg/ml) were sequenced using Illumina and Nanopore sequencing platforms. The proportions of N. gonorrhoeae isolates with resistance were 84.6% for penicillin, 91.5% for tetracycline, and 96.6% for ciprofloxacin. All isolates were susceptible to spectinomycin, azithromycin, cefixime, and ceftriaxone. An adenine deletion within a 13 bp inverted repeat sequence in the mtrR promoter and an H105Y mutation in the mtrR coding region were found in the N. gonorrhoeae isolate with the highest azithromycin MIC value (1 μg/ml). Three high-level penicillin-resistant isolates contained nonmosaic type II penA and had mutations in penB and the mtrR coding region. All isolates with high-level penicillin resistance carried the conjugative plasmids with or without the Dutch type tetM determinant, the beta-lactamase plasmid (Rio/Toronto), and the cryptic plasmid. The gonococcal population in Thailand showed high susceptibility to ceftriaxone and azithromycin, current dual therapy recommended for gonorrhea treatment. As elevated MIC of azithromycin has been observed in 1 strain of N. gonorrhoeae, expanded and enhanced surveillance of antimicrobial susceptibility and study of genetic resistance determinants are essential to improve treatment guidelines.

182. Back to The Future: Increasing Penicillin Susceptibility among Methicillin-Susceptible Staphylococcus aureus Osteoarticular Infections in Children

BackgroundStarting in the late 1940s-1950s Staphylococcus aureus isolates gained resistance to penicillin largely through the acquisition of β-lactamases. In recent years, some centers have described an increase in the proportion of methicillin susceptible S. aureus (MSSA) which are also susceptible to penicillin (PSSA). There are little data on the prevalence or clinical significance of PSSA in children. Acute hematogenous osteoarticular infections (AHOAIs, including osteomyelitis and septic arthritis) are the most common manifestation of invasive S. aureus disease in children. We investigated the prevalence of penicillin susceptibility among MSSA AHOAI isolates at two children’s hospitals.MethodsMSSA AHOAI isolates were obtained through surveillance studies at Texas Children’s (TCH) and St. Louis Children’s Hospitals (SLCH) from 1/2011- 12/2019. All isolates underwent PCR for blaZ β-lactamase, PVL genes and agr group. All blaZ negative isolates then underwent penicillin susceptibility testing using macrobroth dilution. Isolates which were blaZ negative and had a penicillin MIC ≤ 0.125 μg/ml were regarded as PSSA.Results329 unique isolates were available and included in the study. The median patient age was 9.2 years (IQR: 5.1-12.2). Overall, 22 isolates were found to be penicillin susceptible (6.7%). No PSSA isolates were detected prior to 2015 but increased yearly thereafter; by the final study year 20.4% of isolates were PSSA (p=0.001, Figure 1). Patients with PSSA isolates were slightly older than those with resistant isolates (median age 11.8 years vs. 9.1 years, p=0.08) and PSSA were more commonly identified at SLCH (12.9% vs. 5.2%, p=0.04). PSSA were similar to penicillin-resistant isolates in terms agr group and PVL carriage as well as clinical presentation and outcomes. For PSSA, the MIC90 for penicillin (0.06 μg/ml) was much lower than that for other β-lactams (Figure 2). The figure describes the relative frequency of penicillin susceptible S. aureus (PSSA) over time among MSSA osteoarticular infection isolates in children. Distribution of MICs to penicillin, ampicillin, cefazolin, cephalexin and oxacillin among PSSA isolates.ConclusionPSSA appears to be increasing among AHOAI isolates in US children, although geographic variability does occur. Overall, PSSA isolates are associated with a similar clinical presentation as penicillin-resistant isolates. Penicillin susceptibility testing may serve as an avenue for future stewardship intervention in staphylococcal infections. Disclosures Jonathon C. McNeil, MD, Agency for Healthcare Research and Quality (Research Grant or Support)Allergan (Grant/Research Support)Nabriva (Grant/Research Support, Other Financial or Material Support, Site PI for a multicenter trial) Kristina G. Hulten, PhD, Pfizer (Research Grant or Support) Sheldon L. Kaplan, MD, Pfizer (Research Grant or Support)

Antibiotic resistance among invasive Neisseria meningitidis isolates in England, Wales and Northern Ireland (2010/11 to 2018/19).

Invasive meningococcal disease (IMD), caused by Neisseria meningitidis, can have a fatality rate as high as 10%, even with appropriate treatment. In the UK, penicillin is administered to patients in primary care whilst third generation cephalosporins, cefotaxime and ceftriaxone, are administered in secondary care. The first-choice antibiotic for chemoprophylaxis of close contacts is ciprofloxacin, followed by rifampicin. Immunocompromised individuals are often recommended antibiotic chemoprophylaxis and vaccination due to a greater risk of IMD. Resistance to antibiotics among meningococci is relatively rare, however reduced susceptibility and resistance to penicillin are increasing globally. Resistance to third generation cephalosporins is seldom reported, however reduced susceptibility to both cefotaxime and ceftriaxone has been observed. Rifampicin resistance has been reported among meningococci, mainly following prophylaxis, and ciprofloxacin resistance, whilst uncommon, has also been reported across the globe. The Public Health England Meningococcal Reference Unit receives and characterises the majority of isolates from IMD cases in England, Wales and Northern Ireland. This study assessed the distribution of antibiotic resistance to penicillin, rifampicin, ciprofloxacin and cefotaxime among IMD isolates received at the MRU from 2010/11 to 2018/19 (n = 4,122). Out of the 4,122 IMD isolates, 113 were penicillin-resistant, five were ciprofloxacin-resistant, two were rifampicin-resistant, and one was cefotaxime-resistant. Penicillin resistance was due to altered penA alleles whilst rifampicin and ciprofloxacin resistance was due to altered rpoB and gyrA alleles, respectively. Cefotaxime resistance was observed in one isolate which had an altered penA allele containing additional mutations to those harboured by the penicillin-resistant isolates. This study identified several isolates with resistance to antibiotics used for current treatment and prophylaxis of IMD and highlights the need for continued surveillance of resistance among meningococci to ensure continued effective use.

Antibiotic resistance profiles of Staphylococcus spp. from white button mushrooms and handlers

The presence of Staphylococcus spp. has increasingly been reported in food products and poses a public health threat. The aim of this study was to determine the diversity of Staphylococcus spp. and the antibiotic resistance profiles of isolates obtained from freshly harvested and packed ready-to-eat mushrooms (n=432) and handlers’ hands (n=150). A total of 56 Staphylococcus isolates [46.4% (n=26) from hands and 53.6% (n=30) from mushrooms] were recovered belonging to 10 species. Staphylococcus succinus isolates (n=21) were the most prevalent, of which 52.4% came from mushrooms and 47.6% from hands. This was followed by S. equorum isolates [n=12; 91.7% (n=11) from mushrooms and 8.3% (n=1) from hands] and S. saprophyticus [n=9; 66.7% (n=6) from mushrooms and 33.3% (n=3) from hands]. Six isolates that were characterised as multidrug resistant were isolated from hands of handlers. Most (83.9%; n=47) of the 56 isolates were resistant to penicillin [53.2% (n=25) from mushrooms and 46.8% (n=22) from hands] and 14.3% (n=8) were resistant to cephalosporin classes [25% (n=2) from mushrooms and 75% (n=6) from hands], both of which are used to treat staphylococcal infections. Antibiotic resistance genes blaZ [25.0% (n=14) of all isolates of which 71.4% (n=10) were from hands and 28.57% (n=4) from mushrooms], tetL and tetK [both 1.8% (n=1) from hands], mecA [5.4% (n=3) from hands] and ermA [1.8% (n=1) from mushrooms] were detected from the 56 isolates. Only two (25.0%) of the eight methicillin-resistant staphylococci harboured the mecA gene, while only 11 (23%) of the 47 penicillin-resistant isolates harboured the blaZ gene [36.4% (n=4) from mushrooms and 63.6% (n=7) from hands]. Our results demonstrate that food handlers and harvested and packed ready-to-eat mushrooms could be a source of diverse Staphylococcus spp. that exhibit antimicrobial resistance. Clinically relevant S. aureus was only detected on one handler’s hand; however, the isolate was not multidrug resistant. The presence of diverse Staphylococcus spp. on mushrooms and the hands of handlers is a potential public health concern due to their potential to cause opportunistic infections.Significance: This study is the first to describe the antibiotic resistance profiles and antibiotic gene presence of Staphylococcus spp. isolated from fresh mushrooms and hands of pickers and packers. Mushrooms and handlers in this study were demonstrated to be possible routes of transmission of Staphylococcus spp. that are antibiotic resistant and which harbour antibiotic resistance genes, presenting a possible public health hazard.

Vaccination as a solution of the issue of resistance S. pneumoniae

Relevance Antimicrobial resistance (AMR) is a growing acute global health issue. Uncontrolled combined use of antibiotics determines changes in the microbial agents' properties, leading to a decrease of susceptibility to antimicrobial substances and increasing the probability of severe invasive bacterial infections with limited possibilities to treat them. Vaccination against pneumococcal infection as well as improvement of the health care providers awareness can be considered as part of AMR management strategies. A number of epidemiological observations have been published indicating a change in antimicrobial resistance to Streptococcus pneumoniae following the implementation of the national infant immunization program against pneumococcal infection. Aim. To analyze the literature data on the pneumococcal vaccination contribution to the reduction of the number of pneumococcal diseases and the need for antibacterial therapy and to assess the burden of invasive pneumococcal diseases globally and in the Republic of Belarus (RB) and to identify gaps in the awareness of practicing physicians on pneumococcal infection immunoprophylaxis, its' effectiveness and impact on AMR. Conclusions. WHO considers and promote vaccination as an AMR containment measure. There are a lot of publications with the examples of countries with pneumococcal infection pediatric national immunization programs (NIP) and epidemiological data on changes in the S.pneumoniae sensitivity to antibacterial drugs after the start of the NIP. During the period of the mass PCV immunization and the subsequent drift of strains the sensitivity to antimicrobials may return. In Israel 88% and 93% reduction of otitis media cases caused by resistant to penicillin and macrolides vaccine types of S.pneumoniae after the phased introduction of PCV7 and PCV13 to the infant NIP, comparing to the period before vaccination introduction. A decrease in the frequency of bacteremia in combination with a sharp decrease in the proportion of penicillin-resistant pneumococcal isolates (from 50.9% to 5.3%) is also described. In France the proportion of S.pneumoniae strains with reduced susceptibility or resistance to penicillin declined from 47.1% to 39% after the start of PCV13 NIP; researchers also noted a decrease in antibiotic-resistant isolates of S.pneumoniae in children with otitis media: by 26, 31, 55, 29 and 57%, respectively, to penicillin, amoxicillin, cefotaxime, erythromycin and co-trimoxazole to in 2011. A study in Germany revealed a decrease in macrolide-resistant IPD strains after the start of PCV13 use of (in children 8.2%; in adults 8.8%) compared with the period after the introduction of PCV7 (17.3% and 13.0%) and especially in the cohort of children compared with the period before the use of PCV7 (24.8% and 13.3%). In Scotland, there was a decrease in the number of penicillin-resistant strains isolated from the blood of patients with IPD after the introduction of PCV13 into the NIP. The publication of the results of three parallel studies in Finland evaluated the effectiveness of PCV10 in reducing the number of IPD caused by vaccine-specific serotypes of PCV10 by 93%, 98% and 100%, respectively. At the same time, the effectiveness against «related» PCV10 serotypes was equal to 46%, 51% and 78%, respectively, and the estimated effectiveness against unrelated PCV10 serotypes was negative. Composition and resistance profile of the lower respiratory tract microflora isolated from sputum pneumococcal strains in children in 2016–2018, demonstrated up to 72.4% resistance to macrolides, 31.3% resistance to cefotaxime and 8.3% – to ceftriaxone. Spn serotypes structure in under 5 years of age children with meningitis was characterized by relatively high uniformity. Majority (92%) of multiresistant meningeal strains are covered by current PCVs. An anonymous questionnaire showed pediatricians' insufficient awareness about current approaches to pneumococcal infection prevention, underestimation of IPD risks and complications, and the impact of vaccination on reducing AMR. Additional education is required to increase the level of knowledge and understanding of the PCVs role in AMR reduction. PCV NIP introduction will provide additional opportunities to reduce AMR in the Republic of Belarus.

Comparative characterisation of human and ovine non-aureus staphylococci isolated in Sardinia (Italy) for antimicrobial susceptibility profiles and resistance genes.

We present the comparative characterisation of 195 non-aureus staphylococci (NAS) isolates obtained from sheep (n = 125) and humans (n = 70) in Sardinia, Italy, identified at the species level by gap gene polymerase chain reaction (PCR) followed by restriction fragment length polymorphism analysis with AluI. Isolates were tested phenotypically with a disc diffusion method and genotypically by PCR, for resistance to 11 antimicrobial agents including cationic antiseptic agents. Among the ovine isolates, Staphylococcus epidermidis (n = 57), S. chromogenes (n = 29), S. haemolyticus (n = 17), S. simulans (n = 8) and S. caprae (n = 6) were the most prevalent species, while among human isolates, S. haemolyticus (n = 28) and S. epidermidis (n = 26) were predominant, followed by S. lugdunensis and S. hominis (n = 4). Of the 125 ovine isolates, 79 (63.2%) did not carry any of the resistance genes tested, while the remainder carried resistance genes for at least one antibiotic. The highest resistance rates among ovine isolates were recorded against tetracycline (20.8%), and penicillin (15.2%); none was resistant to methicillin and two exhibited multidrug resistance (MDR); one of which was positive for the antiseptic resistance smr gene. By contrast, most human isolates (59/70, 84.3%) were resistant to ⩾1 antimicrobials, and 41 (58.6%) were MDR. All 52 (74.3%) penicillin-resistant isolates possessed the blaZ gene, and 33 of 70 (47.1%) harboured the mec gene; of these, seven were characterised by the Staphylococcal Chromosomal Cassette (SCCmec) type IV, 6 the type V, 5 of type III and one representative each of type I and type II. The majority (57.1%) was erythromycin-resistant and 17 isolates carried only the efflux msrA gene, 11 the methylase ermC gene and an equal number harboured both of the latter genes. Moreover, 23 (32.8%) were tetracycline-resistant and all but one possessed only the efflux tetK gene. qacA/B and smr genes were detected in 27 (38.6%) and 18 (25.7%) human NAS, respectively. These results underline a marked difference in species distribution and antimicrobial resistance between ovine and human-derived NAS.

Virulence factors and antimicrobial resistance in Staphylococcus aureus isolated from bovine mastitis in Brazil.

This study aimed to evaluate virulence factors and genetic markers of antimicrobial resistance in 400 Staphylococcus aureus strains isolated from bovine mastitis in four Brazilian states, as well as to assess the association between these characteristics and field information. Virulence factors and drug resistance genes were identified by PCR screening. Biofilm-forming and hemolytic phenotype were detected using Congo red Tryptic Soy Broth and defibrinated sheep blood agar, respectively. Of all isolates, 83.5% were biofilm-forming and 98.5% strains exhibited biofilm gene icaAD, and a significant association between phenotype and genotype for biofilm was observed (P = 0.0005). Hemolysin genes were observed in 82.85% (hla+hlb+), 16.5% (hla+) and 0.75% (hlb+) isolates, whereas the hemolytic phenotype exhibited was complete and incomplete hemolysis in 64.25%, complete in 28.25%, incomplete in 4.75%, and negative in 2.75% of the strains. Virulence factors genes luk, seb, sec, sed, and tst were observed in 3.5%, 0.5%, 1%, 0.25%, and 0.74% isolates, respectively. The gene blaZ was detected in 82.03% of penicillin-resistant isolates, whereas tetK and aac(6')-Ie-aph(2')-Ia were observed in 33.87% and 45.15% of the tetracycline and aminoglycosides-resistant isolates, respectively. Fluoroquinolone resistance gene mepA was detected for the first time in S. aureus from bovine mastitis. Resistance genes tetM (3.22%), tetL (1.61%), ermA (14.29%), ermB (14.29%), ermC (33.3%), ermT (9.52%), ermY (4.76%), msrA (9.52%), and mphC (9.52%) were also detected among resistant isolates. No association between virulence factors or antimicrobial-resistant genes and year of isolation, geographic origin, or antimicrobial resistance profile was observed. Our results showed that S. aureus strains isolated from bovine mastitis in the four Brazilian states sampled are mainly biofilm-forming and hemolytic, whereas virulence genes associated with enterotoxins, luk and tst, were less frequently observed. Moreover, a wide variety of resistance genes that confer resistance to almost all classes of antimicrobial agents approved for use in animals and humans were found. Overall, the data point to a great pathogenic potential of S. aureus associated with bovine mastitis and to the non-negligible risks to public health of staphylococcal infections from animal origin.