Increase in Penicillin and multidrug resistance in Streptococcus pneumoniae (1993-2016): report from a tertiary care hospital laboratory, Pakistan

Background: Streptococcus pneumoniae is a major cause of morbidity and mortality worldwide. With the emergence of penicillin-resistant S. pneumoniae (PRSP), treatment has become challenging. The Clinical and Laboratory Standards Institute (CLSI) in 2008 revised its guidelines for S. pneumoniae and recommended separate penicillin breakpoints for meningeal and non-meningeal strains. Similar to penicillin’s, resistance to other classes of antibiotics has emerged globally. Objective: The objective of this study is to determine the trend of resistance to antimicrobials in S. pneumoniae infections and the impact of new CLSI guidelines on penicillin susceptibility among meningeal isolates. Methodology: Twenty-four years (1993-2016) data from S. pneumoniae isolates and their antimicrobial susceptibility was retrieved from the computerized database. Data was divided into two groups for analysis, pre-2008 and post 2008. Results: Penicillin resistance remained unchanged in non-meningeal isolates during both study periods. A significant rise in penicillin resistance in meningeal isolates was observed in the second period 2008-2016 (2.9% vs 36.2%). High resistance rates were observed for co-trimoxazole, tetracycline and erythromycin. Increased trend of multi-drug resistant (MDR) strains were also noted, from 11% in 1999 to 36% in 2016. Conclusion: The emergence of MDR strains is evident from our dataset. It seems like the rise in PRSP in meningeal isolates is due to revised CLSI guidelines. Overall low resistance to penicillin in non-meningeal isolates and no resistance to ceftriaxone is encouraging and will assist in drafting local guidelines. Cautious use of antimicrobials are essential to reduce further emergence of antimicrobial resistance in indigenous isolates.

Occurrence and Antimicrobial Susceptibility Profiles of Streptococcus equi subsp. zooepidemicus Strains Isolated from Mares with Fertility Problems

Streptococcus equi subsp. zooepidemicus (S. zooepidemicus), is a β-hemolytic Streptococcus belonging to the Lancefield group C; it is a rare human pathogen, but in horses, it is frequently associated with endometritis. This study aimed to isolate S. zooepidemicus strains, associated with bacterial endometritis in mares, and to define their antimicrobial resistance profile. Twenty-three isolates were recovered from one hundred ninety-six equine uterine swabs (11.7%). Bacterial identification was carried out by Api 20 Strep and confirmed by matrix assisted laser desorption ionization time of flight mass spectrometry (MALDI-TOF-MS), while antimicrobial susceptibility testing was performed by disk diffusion method on Muller Hinton agar plates. The antibiotic resistance profiles of the isolates revealed a high percentage of resistance to amikacin (95.6%), ampicillin (73.9%) and tetracycline (69.6%), while ceftiofur and ceftriaxone were highly effective with 82.6% and 78.3% of the isolates inhibited, respectively. An intriguing value of resistance to penicillin (34.8%), which represents the first-choice antibiotic in equine S. zooepidemicus infections, was observed. Furthermore, a high prevalence of multidrug-resistant strains (82.6%) was recorded. Continuous surveillance of this potential zoonotic pathogen and an appropriate antimicrobial stewardship program with the promotion of correct use of antimicrobials, after a proper diagnosis, are needed to allow an effective therapy.

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.

Molecular epidemiology of methicillin resistant Staphylococcus species in healthcare workers of a blood bank in the Brazilian Amazon

Background Healthcare workers are susceptible to colonization by multiresistant bacteria, which can increase the risk of outbreaks. Methods Samples were collected from the nasopharynx, hands, and lab coats of healthcare workers. The phenotypic identification was carried out using a VITEK®2 rapid test system. PCR tests for the mecA gene and the sequencing of the amplicons were performed. Staphylococcus epidermidis and Staphylococcus aureus phylogenies were reconstructed using the Bayesian inference. Results A total of 225 healthcare workers participated in this study. Of these, 21.3% were male and 78.7% female. S. epidermidis and S.aureus showed high levels of resistance to penicillin, ampicillin, erythromycin, tetracycline and cefoxitin. The prevalence of methicillin resistant S. aureus was 3.16% and methicillin resistant S. epidermidis was 100%. Multilocus sequence typing identified 23 new S. epidermidis sequence types, and one new allele and sequence type for S. aureus. The frequency of methicillin-resistant S. epidermidis in nursing and hemotherapy technicians as a percentage of the total number of healthcare workers was 5.8–3.1%, while the frequency of methicillin resistant S. aureus in hemotherapy technicians and biomedics, as a percentage of the total number of healthcare workers was 4.2–8.9%%. Conclusions The healthcare workers at the city’s blood bank, even when taking the necessary care with their hands, body and clothes, harbour methicillin-resistant S. aureus and S. epidermidis sequence types, which, as a potential source of multidrug resistant bacteria, can contribute to nosocomial infections among hematological patients.

A survey of prevalence and phenotypic and genotypic assessment of antibiotic resistance in Staphylococcus aureus bacteria isolated from ready-to-eat food samples collected from Tehran Province, Iran

Background Resistant Staphylococcus aureus (S. aureus) bacteria are considered among the major causes of foodborne diseases. This survey aims to assess genotypic and phenotypic profiles of antibiotic resistance in S. aureus bacteria isolated from ready-to-eat food samples. Methods According to the previously reported prevalence of S. aureus in ready-to-eat food samples, a total of 415 ready-to-eat food samples were collected from Tehran province, Iran. S. aureus bacteria were identified using culture and biochemical tests. Besides, the phenotypic antibiotic resistance profile was determined by disk diffusion. In addition, the genotypic pattern of antibiotic resistance was determined using the PCR. Results A total of 64 out of 415 (15.42%) ready-to-eat food samples were contaminated with S. aureus. Grilled mushrooms and salad olivieh harbored the highest contamination rate of (30%), while salami samples harbored the lowest contamination rate of 3.33%. In addition, S. aureus bacteria harbored the highest prevalence of resistance to penicillin (85.93%), tetracycline (85.93%), gentamicin (73.43%), erythromycin (53.12%), trimethoprim-sulfamethoxazole (51.56%), and ciprofloxacin (50%). However, all isolates were resistant to at least four antibiotic agents. Accordingly, the prevalence of tetK (70.31%), blaZ (64.06%), aacA-D (57.81%), gyrA (50%), and ermA (39.06%) was higher than that of other detected antibiotic resistance genes. Besides, AacA-D + blaZ (48.43%), tetK + blaZ (46.87%), aacA-D + tetK (39.06%), aacA-D + gyrA (20.31%), and ermA + blaZ (20.31%) were the most frequently identified combined genotypic patterns of antibiotic resistance. Conclusion Ready-to-eat food samples may be sources of resistant S. aureus, which pose a hygienic threat in case of their consumption. However, further investigations are required to identify additional epidemiological features of S. aureus in ready-to-eat foods.

Hygienic-sanitary quality of ready-to-eat salmon sashimi (Salmo Salar)

Fifty sashimi samples from 5 restaurants were characterized for enumeration of Aerobic Psychrotrophic Heterotrophic Bacteria (APHB) and Aerobic Mesophilic Heterotrophic Bacteria (AMHB), Enterobacteriaceae, coliforms at 45ºC (thermotolerant coliforms), coagulase-positive Staphylococci, presence of Vibrio parahaemolyticus and Salmonella sp., and determination of hydrogen potential (pH) and total volatile basic nitrogen (TVB-N). The bacterial isolates were evaluated for their resistance profile to the antimicrobial agents Penicillin, Ampicillin, Cefoxitin, Cefotaxime, Amikacin, Gentamicin, Tetracycline, and Trimethropim-sulfamethoxazole. The results showed that APHB, AMHB and coagulase-positive Staphylococci counts and the determination of pH and TVB-N were in accordance with national and international standards adopted as safe limits for consumption. In contrast, the Enterobacteriaceae and thermotolerant coliforms counts and presence of Salmonella sp. and V. parahaemolyticus were in disagreement with those standards, raising concern about the hygienic-sanitary quality of sashimi. The Staphylococcus aureus and V. parahaemolyticus isolates showed resistance to Penicillin, Ampicillin, Cefoxitin, Cefotaxime, Tetracycline, Gentamicin, and Amikacin, while the Salmonella sp. isolate showed no resistance to all the antimicrobial agents studied. The results showed that 48% of the samples were fit for consumption while 52% had unsatisfactory hygienic-sanitary quality for the parameters evaluated.

A Study on High and Low level Drug Resistance Pattern Among Clinical Isolates of Enterococci .

Introduction: The combined abilities of colonisation and both inherent and acquired resistance have made Enterococci a significant human pathogen. Aims and Objectives: This study was done to determine the Minimum Inhibitory Concentration of various antibiotics against Enterococci and to correlate the phenotypic and genotypic characteristics of Enterococci with low level and high level drug resistance. Materials and Methods: A total of 774 isolates of Enterococci obtained from various clinical samples were subjected to antimicrobial susceptibility testing by Kirby Bauer Disk Diffusion method. The Minimum Inhibitory Concentration of various antibiotics were determined by Vitek 2 automated system, agar dilution and E test. Results: 15 out of 774 isolates showed the presence of vancomycin resistant genes by Multiplex PCR. 10 (90.91 %) isolates out of 11 E. faecalis with van A gene showed high level resistance to Penicillin (16-64 µg/ml). 8 (72.73 %) out of 11 isolates showed high level resistance to Gentamicin (512-1024 µg/ml). 6 (54.55 %) , out of 11 isolates were resistant to β lactams. One isolate of E. faecalis from urine with van B gene showed showed high level resistance to Penicillin (32 µg/ml), Linezolid (≥ 8µg/ml), high level resistance to Gentamicin (1024 µg/ml), Fluoroquinolones (≥ 8µg/ml) and Macrolides (≥ 8µg/ml). Conclusion: Isolates of Enterococci resistant to glycopeptides, penicillin, Betalactams and aminoglycosides have important clinical implications in the treatment for infection.

Molecular Characterization and Survive Abilities of Salmonella Heidelberg Strains of Poultry Origin in Brazil

The aim of the study was to evaluate the genotypic and phenotypic characteristics of 20 strains of S. Heidelberg (SH) isolated from broilers produced in southern Brazil. The similarity and presence of genetic determinants linked to virulence, antimicrobial resistance, biofilm formation, and in silico-predicted metabolic interactions revealed this serovar as a threat to public health. The presence of the ompC, invA, sodC, avrA, lpfA, and agfA genes was detected in 100% of the strains and the luxS gene in 70% of them. None of the strains carries the blaSHV, mcr-1, qnrA, qnrB, and qnrS genes. All strains showed a multidrug-resistant profile to at least three non-β-lactam drugs, which include colistin, sulfamethoxazole, and tetracycline. Resistance to penicillin, ceftriaxone (90%), meropenem (25%), and cefoxitin (25%) were associated with the presence of blaCTX–M and blaCMY–2 genes. Biofilm formation reached a mature stage at 25 and 37°C, especially with chicken juice (CJ) addition. The sodium hypochlorite 1% was the least efficient in controlling the sessile cells. Genomic analysis of two strains identified more than 100 virulence genes and the presence of resistance to 24 classes of antibiotics correlated to phenotypic tests. Protein-protein interaction (PPI) prediction shows two metabolic pathways correlation with biofilm formation. Virulence, resistance, and biofilm determinants must be constant monitoring in SH, due to the possibility of occurring infections extremely difficult to cure and due risk of the maintenance of the bacterium in production environments.

Comparison of endocervical swabs to cultured isolates for the detection of antimicrobial resistance determinants in Neisseria gonorrhoeae

Background: The global emergence of antimicrobial resistance (AMR) in Neisseria gonorrhoeae to various antibiotics is a public health concern. To date, there have been no published South African studies that have compared the primary swab to the cultured isolates for the detection of N. gonorrhoeae AMR determinants. This study provides data on such a comparison. Methods: Paired endocervical swabs were collected from 307 pregnant women. The first swab was stored in an Amies charcoal transport media for culture assessment and the second swab was used for the molecular detection of resistant determinants. Specific targets (genes/plasmids/mutations) associated with resistance to penicillin, tetracycline, ciprofloxacin, spectinomycin, cefixime, azithromycin and ceftriaxone were detected from both the cultured isolates and the endocervical swabs. Results: Of the 307 samples tested in this study, only six samples tested positive for culture. A total of 24 samples tested positive for N. gonorrhoeae with the quantitative polymerase chain reaction (qPCR) assay. The six samples which tested positive for culture fell within the qPCR positives group. Since this study was designed to directly compare the culture swabs to the endocervical swabs for the detection of AMR determinants, the current analysis included only the six culture samples and six paired endocervical swab samples (n = 6). All six isolates were resistant to tetracycline and penicillin G while five of the six isolates were resistant to ciprofloxacin. All isolates were susceptible to the remaining antimicrobials. There was a 100% correlation between the cultured isolates and endocervical swabs for detecting the specific AMR determinants, conferring resistance to tetracycline, penicillin G and ciprofloxacin. Conclusion: Based on the findings of this study, tracking emerging patterns of resistance from the molecular level using only the endocervical swabs may serve as an attractive future research direction.

Phenotypic and Molecular Traits of Staphylococcus coagulans Associated with Canine Skin Infections in Portugal

Staphylococcus coagulans is among the three most frequent pathogens of canine pyoderma. Yet, studies on this species are scarce. Twenty-seven S. coagulans and one S. schleiferi, corresponding to all pyoderma-related isolations from these two species at two veterinary laboratories in Lisbon, Portugal, between 1999 and 2018 (Lab 1) or 2018 (Lab 2), were analyzed. Isolates were identified by the analysis of the nuc gene and urease production. Antibiotic susceptibility towards 27 antibiotics was evaluated by disk diffusion. Fourteen antibiotic resistance genes were screened by PCR. Isolates were typed by SmaI-PFGE. Two S. coagulans isolates (2/27, 7.4%) were methicillin-resistant (MRSC, mecA+) and four (4/27, 14.8%) displayed a multidrug-resistant (MDR) phenotype. We observed resistance to penicillin (17/27, 63.0%), fluoroquinolones (11/27, 40.7%), erythromycin and clindamycin (3/27, 11.1%), fusidic acid (3/27, 11.1%) and tetracycline (1/27, 3.7%). The blaZ and erm(B) genes were carried by 16 and 1 isolates resistant to penicillin and erythromycin/clindamycin, respectively. Only three S. coagulans carried plasmids. The single S. schleiferi isolate presented an MDR phenotype. SmaI-PFGE revealed a limited genetic diversity of S. coagulans, with a predominant lineage present from 2001 to 2018. This study describes the first MRSC causing canine infection in Portugal and reveals a high burden of antimicrobial resistance, with the emergence of MDR phenotypes within the main lineages.