Nocardiosis in the Illawarra-Shoalhaven region from 2010 to 2019.

There is a need for simple microbiology diagnostics to enable antimicrobial resistance surveillance in low- and middle-income countries.
To investigate the field utility of InTray COLOREX plates for urine culture and ESBL detection.
Clinical urine samples from Mahosot Hospital, Vientiane, Lao PDR were inoculated onto chromogenic media and InTray COLOREX Screen plates between June and August 2020.
Urine and isolates from other clinical specimens were inoculated onto COLOREX ESBL plates. A simulated field study investigating the field utility of the InTray COLOREX plates was also completed.
 In total, 355 urine samples were inoculated onto standard chromogenic agar and InTray COLOREX Screen plates, and 154 urine samples and 54 isolates from other clinical specimens on the COLOREX ESBL plates.
Growth was similar for the two methods (COLOREX Screen 41%, standard method 38%) with 20% discordant results, mainly due to differences in colony counts or colonial appearance.
Contamination occurred in 13% of samples, with the COLOREX Screen plates showing increased contamination rates, potentially due to condensation.
ESBL producers were confirmed from 80% of isolates from the COLOREX ESBL plates, and direct plating provided rapid detection of presumptive ESBL producers. Burkholderia pseudomallei also grew well on the ESBL plates, a relevant finding in this melioidosis-endemic area.
The InTray COLOREX Screen and ESBL plates were simple to use and interpret, permitting rapid detection of uropathogens and ESBLs, and have the potential for easy transport and storage from field sites and use in laboratories with low capacity.

UK standards for microbiology investigations of ear infection (SMI B1) are inadequate for the recovery of fungal pathogens and laboratory diagnosis of otomycosis: a real-life prospective evaluation.

 We evaluated the recovery of fungal pathogens from clinical external ear samples from patients with otitis externa (OE) using the UK national Standard Microbiology Investigations of ear infection (SMI B1).
The UK SMI B1 protocol including a single Sabouraud dextrose agar with chloramphenicol (SABC) incubated at 37°C for 48 hours was compared to a standard fungal-specific culture method using two SABC agar plates incubated at 28°C and 37°C for 2 weeks with an extra Candida chromogenic agar incubated at 37°C for 5 days.
This was undertaken on ear samples from patients with OE from January 2020 to December 2020.
 Altogether, 304 individual patient ear swabs were prospectively examined.
The positivity rate of UK standard was 14% (42/304) versus 26% (79/304) for the fungal-specific protocol (p<0.05).
The standard protocol identified seven compared to 17 species using the fungal-specific protocol. A total of 93 fungal isolates were recovered; nine different yeasts and eight filamentous fungal species.
Candida parapsilosis (38/304; 13%), C. albicans (10/304; 3%) and C. orthopsilosis (6/304; 2%) were common yeast species. Aspergillus niger complex (16/304; 5%) was the most common mold, followed by A. fumigatus complex (3/304; 1%).
Many less common and emerging yeasts and molds were only isolated from samples cultured using a fungal-specific protocol.
 Our results suggest that the UK SMI B1 media and procedures are inadequate to detect all fungal agents causing otomycosis.
Fungal-specific culture protocols increase the recovery rate and diversity of fungal pathogens isolated from ear samples.

Comparative Genomics of Borderline Oxacillin-Resistant Staphylococcus aureus Detected during a Pseudo-outbreak of Methicillin-Resistant S. aureus in a Neonatal Intensive Care Unit.

  1. Active surveillance for methicillin-resistant Staphylococcus aureus (MRSA) is a component of our neonatal intensive care unit (NICU) infection prevention efforts.
  2. Recent atypical trends prompted review of 42 suspected MRSA isolates. Species identification was confirmed by matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS), and methicillin resistance was reevaluated by PBP2a lateral flow assay, cefoxitin/oxacillin susceptibility testing, mecA and mecC PCR, and six commercially available MRSA detection agars.
  3. All isolates were confirmed S. aureus, but only eight were MRSA (cefoxitin resistant, PBP2a positive, mecA positive, growth on all MRSA screening agars). One MRSA isolate was cefoxitin susceptible but PBP2a and mecA positive, and the remaining 33 were cefoxitin susceptible, PBP2a negative, and mecA negative; interestingly, these isolates grew inconsistently across MRSA screening agars and had susceptibility profiles consistent with that of borderline oxacillin-resistant S. aureus (BORSA).
  4. Comparative genomic analyses found these BORSA isolates to be phylogenetically diverse and not representative of clonal expansion or shared gene content, though clones of two NICU strains were infrequently observed over 8 months.
  5. We identified 6 features-substitutions and truncations in PBP2, PBP4, and GdpP and beta-lactamase hyperproduction-that were used to generate a random forest classifier to distinguish BORSA from methicillin-susceptible S. aureus (MSSA) in our cohort.
  6. Our model demonstrated a robust ability to predict the BORSA phenotype among isolates collected across two continents (validation area under the curve [AUC], 0.902). Taking these findings together, we observed an unexpected prevalence of BORSA in our NICU, BORSA misclassification by existing MRSA screening methods, and markers that are together discriminatory for BORSA and MSSA within our cohort. This work has implications for epidemiological reporting of MRSA rates for centers using different screening methods.
  7. In this study, we found a high prevalence of Staphylococcus aureus isolates exhibiting a borderline oxacillin resistance phenotype (BORSA) in our neonatal intensive care unit (NICU) serendipitously due to the type of MRSA screening agar used by our laboratory for active surveillance cultures.
  8. Subsequent phenotypic and molecular characterization highlighted an unexpected prevalence and variability of BORSA isolates. Through whole-genome sequencing, we interrogated core and accessory genome content and generated a random forest classification model to identify mutations and truncations in the PBP2, PBP4, and GdpP proteins and beta-lactamase hyperproduction, which correlated with BORSA and MSSA phenotypes among S. aureus clinical isolates collected across two continents.
  9. In consideration of these findings, this work will help clinical microbiology laboratories and clinicians identify MRSA screening shortfalls and draw attention to the non-mecA-mediated BORSA phenotype.

nvestigation of Candidal Species among People Who Suffer from Oral Potentially Malignant Disorders and Oral Squamous Cell Carcinoma.

 The aim of the present study was to evaluate the candidal species among masses with oral potentially malignant disorders and oral squamous cell carcinoma.
  •  This prospective and observational study was conducted by the Department of Oral Pathology and Microbiology, S. B. Patil Dental College, Bidar, Karnataka, India, from February 2018 to January 2019. The study composed of total of 150 individuals, of which 50 individuals did not had any visible manifestations, 50 were analyzed with potentially malignant disorders (PMDs) in particular oral leukoplakia, oral lichen planus, and oral submucous fibrosis and last group of 50 individuals were suffering from oral squamous cell carcinoma (OSCC).
  • First, the swab samples were elicited from culture technique after that incisional biopsy of the discernible investigated lesion was done for the purpose of justopathological verification. The swab samples were streak on Sabouraud dextrose agar (SDA) and HiCrome Candida Differential HiVeg agar/CHROMagar medium and incubation at 37°C for 24-48 h. Biopsy was done for all the samples.
  •  The proportion of candidates as men and women in control was 45 (90%) and 5 (10%), in PMD was 30 (60%) and 20 (40%), and in OSCC was 45 (90%) and 5 (10%), correspondingly. On evaluation on SDA medium in controls, PMD and OSCC groups, Candida was founded in 14 (28%), 20 (40%), and 42 (84%) and not founded in 36 (72%), 30 (60%), and 8 (16%) folks, subsequently.
  • Intragroup contrast illustrated exceedingly necessary distinction with P = 0.000 between both controls versus OSCC and PMD in comparison to OSCC. Nevertheless, controls versus PMD manifested insignificant, P = 0.119. Investigation on CHROM AGAR media among controls, PMD and OSCC groups, Candida species was seen in 11 (22%), 19 (38%), and 40 (80%) and absent in 39 (78%), 31 (62%), and 10 (20%) individuals, respectively. On statistical inspection, the variations noted were enormous, (P = 0.000).
On speciation of Candida in CHROM agar among the controls, PMD and OSCC groups, Candida albicans species was present in 9 (18%), 16 (32%), and 6 (12%), Candida krusei in 3 (6%), 6 (12%), and 13 (26%), Candida glabrata in 0, 0, and 8 (16%), and Candida tropicalis in 0, 0, and 3 (6%) cases, respectively.

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Nonetheless, only OSCC group reveal amalgamation of species such as C. glabrata and C. krusei was present in 2 (4%) case, C. tropicalis and C. krusei in was present 3 (6%) cases, C. tropicalis and C. glabrata was present in 2 (4%) case, C. albicans and C. tropicalis was present in 2 (4%) cases, and C. kruseiC. glabrata with C. albicans was present in 1 (2%) case, respectively.
All other types of fungi were regarded as infectious excluding Candida, on analysis on SDA medium, infestation in the form of fungal molds was seen in 18 (36%) in controls, 12 (24%) in PMD and 8 (16%) in OSCC groups.
We interpreted that the chief carrier of candidal species in PMDs and OSCC, yet more light is to be thrown on the topic that Candida has particular establishment in PMDs or in malignancy.

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