Clinical presentation, antimicrobial resistance, and treatment outcomes of Aeromonas human infections: A 14-year retrospective study and comparative genomics of two isolates from fatal cases.

BACKGROUND: Aeromonas virulence may not be entirely dependent on the host immune status. Pathophysiologic determinants of disease progression and severity remain unclear. METHODS: One hundred five patients with Aeromonas infections and 112 isolates were identified, their clinical presentations and outcomes analyzed, and their antimicrobial resistance (AMR) patterns assessed. Two isolates (A and B) from fatal cases of Aeromonas dhakensis bacteremia were characterized using whole genome sequence analysis. Virulence factor- and AMR-encoding genes from these isolates were compared with a well-characterized diarrheal isolate A. dhakensis SSU, and environmental isolate A. hydrophila ATCC_7966T. RESULTS: Skin and soft tissue infections, traumatic wound infections, sepsis, burns, and intraabdominal infections were common. Diabetes, malignancy, and cirrhosis were frequent comorbidities. Male sex, age ≥ 65 years, hospitalization, burns, and intensive care were associated with complicated disease. High rates of AMR to carbapenems and piperacillin-tazobactam were found. Treatment failure was observed in 25.7% of cases. Septic shock and hospital-acquired infections were predictors of treatment failure. All four isolates harbored assorted broad-spectrum AMR genes including blaOXA, ampC, cphA, and efflux pumps. Only clinical isolates possessed both polar and lateral flagellar genes, genes for various surface adhesion proteins, type 3- and -6 secretion systems and their effectors, and toxin genes, including exotoxin A. Both isolates A and B were resistant to colistin and harbored the mobile colistin resistance-3 (mcr-3) gene. CONCLUSIONS: Empirical therapy tailored to local Aeromonas antibiograms may facilitate more favorable outcomes, while advanced diagnostic methods may aid in identifying correct Aeromonas spp. of significant clinical importance.

The Effect of Povidone-Iodine Irrigation on the Efficacy of Antibiotic Cement In Vitro.

Povidone-iodine is a common antiseptic demonstrating success in reducing infection rates in primary arthroplasty; however, recent data suggest that its use in revision arthroplasty may increase infection rates. This study evaluated the effect of povidone-iodine solution on antibiotic cement and investigated the connection between povidone-iodine and increased infection rates in revision arthroplasty. Sixty antibiotic cement samples (ACSs) were formed using gentamicin-impregnated cement. The ACSs were divided into three groups: group A (n=20) was subject to a 3-minute povidone-iodine soak followed by a saline rinse; group B (n=20) underwent a 3-minute saline soak; and group C (n=20) underwent only a saline rinse. The antimicrobial activity of the samples was tested using a Kirby-Bauer-like assay using Staphylococcus epidermidis. The zone of inhibition (ZOI) was measured every 24 hours for 7 days. All groups possessed the greatest antimicrobial activity at 24 hours. Group C displayed a mass-corrected ZOI of 395.2 mm/g, which was statistically greater than the group B ZOI (313.2 mm/g, P<.05) but not the group A ZOI (346.5 mm/g, P>.05). All groups demonstrated a decrease in antimicrobial activity at 48 through 96 hours, with no significant difference at any time point. Prolonged soaking of antibiotic cement in a povidone-iodine or saline solution results in elution of the antibiotic into the irrigation solution, blunting initial antibiotic concentration. When using antibiotic cement, antiseptic soaks or irrigation should be focused prior to cementation. [Orthopedics. 2024;47(1):10-14.].

Vacuoles in the Breast: A Histologic Clue for an Unusual Presentation of an Atypical Organism.

Infections with nontuberculous mycobacteria (NTM) are increasing in prevalence worldwide, and this group of organisms is emerging as significant clinical pathogens. We present a case of a 58-year-old female with persistent furuncles of the breast who was found to have an NTM infection. This case is unique for the lack of risk factors for NTM in the patient's history, the location of the infection in the breast, and the close cooperation needed across disciplines to arrive at the diagnosis. This multi-disciplinary discussion considers the classic clinical presentation of NTM, it is a characteristic morphological appearance on histopathology, the differential diagnosis, treatment, and the ultimate outcome of the case. This case report and discussion will assist both clinicians and pathologists in the diagnosis of this important infectious disease.

Ovine model of burn wounds grafted with ovine cadaver skin.

Shortage in autograft to cover burn wounds involves a frequent use of cadaver skin (CS) as a temporary cover to prevent infection, dehydration and preparation of wounds for subsequent autografting. We aimed to establish an ovine model of burn wound healing using ovine CS (OCS). Quality and efficacy of fresh and frozen OCS overlaid on to excised 3rd degree flame burn wounds in sheep were evaluated in comparison to autograft. Histologically, autografted wounds maintained normal skin structure at different time points. Wounds overlaid with fresh OCS graft showed signs of rejection starting from day 7. At day 14, the epidermis was mostly rejected. The rejection was completed by day 20 with signs of immunoreaction and presence of many immune cells. Frozen OCS was rejected in the same pattern. Immediately prior to grafting, the thickness was comparable between freshly prepared and frozen OCS for 10 or 40 days. Significant reduction in viability was detected in OCS frozen for 40 days. Both fresh or frozen ovine OCS were rejected within 10 days that mimics CS rejection time in humans (∼8.4 days), suggesting that ovine model of burn wound grafted with OCS can successfully be used in burn wound research mimicking clinical scenario.

Outbreak of coronavirus disease 2019 (COVID-19) among operating room staff of a tertiary referral center: An epidemiologic and environmental investigation.

OBJECTIVE: Investigate an outbreak of coronavirus disease 2019 (COVID-19) among operating room staff utilizing contact tracing, mass testing for severe acute respiratory coronavirus virus 2 (SARS-CoV-2), and environmental sampling. DESIGN: Outbreak investigation. SETTING: University-affiliated tertiary-care referral center. PATIENTS: Operating room staff with positive SARS-CoV-2 molecular testing. METHODS: Epidemiologic and environmental investigations were conducted including contact tracing, environmental surveys, and sampling and review of the operating room schedule for staff-to-staff, staff-to-patient, and patient-to-staff SARS-CoV-2 transmission. RESULTS: In total, 24 healthcare personnel (HCP) tested positive for SARS-CoV-2, including nurses (29%), surgical technologists (25%), and surgical residents (16%). Moreover, 19 HCP (79%) reported having used a communal area, most commonly break rooms (75%). Overall, 20 HCP (83%) reported symptomatic disease. In total, 72 environmental samples were collected from communal areas for SARS-CoV-2 genomic testing; none was positive. Furthermore, 236 surgical cases were reviewed for transmission: 213 (90%) had negative preoperative SARS-CoV-2 testing, 21 (9%) had a positive test on or before the date of surgery, and 2 (<1%) did not have a preoperative test performed. In addition, 40 patients underwent postoperative testing (mean, 13 days to postoperative testing), and 2 returned positive results. Neither of these 2 cases was linked to our outbreak. CONCLUSIONS: Complacency in infection control practices among staff during peak community transmission of SARS-CoV-2 is believed to have driven staff-to-staff transmission. Prompt identification of the outbreak led to rapid interventions, ultimately allowing for uninterrupted surgical service.

Mycobacterium haemophilum and Histoplasma capsulatum coinfection in a renal transplant patient.

We report the case of a 22-year-old man who presented with a Mycobacterium haemophilum and Histoplasma capsulatum coinfection occurring 21 years after a living-donor-related renal transplant.

Direct RT-PCR amplification of SARS-CoV-2 from clinical samples using a concentrated viral lysis-amplification buffer prepared with IGEPAL-630.

The pandemic of 2019 caused by the novel coronavirus (SARS-CoV-2) is still rapidly spreading worldwide. Nucleic acid amplification serves as the gold standard method for confirmation of COVID-19 infection. However, challenges faced for diagnostic laboratories from undeveloped countries includes shortage of kits and supplies to purify viral RNA. Therefore, it is urgent to validate alternative nucleic acid isolation methods for SARS-CoV-2. Our results demonstrate that a concentrated viral lysis amplification buffer (vLAB) prepared with the nonionic detergent IGEPAL enables qualitative detection of SARS-CoV-2 by direct Reverse Transcriptase-Polymerase Chain Reaction (dRT-PCR). Furthermore, vLAB was effective in inactivating SARS-CoV-2. Since this method is inexpensive and no RNA purification equipment or additional cDNA synthesis is required, this dRT-PCR with vLAB should be considered as an alternative method for qualitative detection of SARS-CoV-2.

Molecular characterization of Mycobacterium tuberculosis isolates from Izmir, Turkey.

In recent years, molecular typing methods have been used in epidemiologic studies of Mycobacterium tuberculosis isolates in various areas of the world. However, there have been few data on this issue in Turkey. We describe the molecular characterization of 56 Mycobacterium tuberculosis isolates recovered from individual patients in Izmir and the surrounding area by three different molecular methods. Isolated M. tuberculosis strains were characterized by IS6110 RFLP, spoligotyping and major genetic group designation. In total, 51 RFLP and 35 spoligopatterns were identified. Fourteen (25%) isolates were indicated as low copy number. Based on three genotypic characterization methods together, five clusters with two isolates each were identified. Most of the isolates (98.2%) were assigned as genetic groups 2 or 3. Only one isolate was identified as Beijing family strain (principal genetic group 1). The shared international clades were found to be Beijing-family, var T1 (ST 37), LAM (Latin-American-Mediterranean) 7 (ST 41), LAM 9 (ST 42), Haarlem 1 (ST 47), Haarlem 3 (ST 50) and T1 (ST 53). In this study, IS6110 RFLP, spoligotyping and major genetic group designation were found to be useful methods for molecular epidemiologic studies.

Correlation of microbiologic culture and fine-needle aspiration cytology: A 14-year experience at a single institution.

BACKGROUND: Fine-needle aspiration (FNA) is an important tool for the diagnosis of infectious disease. FNA material should be appropriately submitted for cultures when indicated by preliminary findings. Correlation of cytologic diagnoses with culture results are important quality assurance tools. The current study reviewed 14 years of FNA-culture correlation. METHODS: FNA cytology-culture correlation records from the years 1996 through 2007 and 2010 through 2011 were retrieved from electronic databases compiled for histology and culture correlation. Correlation was limited to those cases for which material was submitted for culture from the FNA sample. Culture results were retrieved from the laboratory or hospital information system. RESULTS: Correlative data included 770 cases. Cytology, culture, or both were positive for microbes in 416 of 770 samples (54%), excluding cultured bacterial skin contaminants. Among the 204 bacteria cases, 93 (46%) were identified by cytology and culture, 92 (45%) were identified by culture only, and 19 (9%) were identified by cytology only. Among the 16 cases of Actinomycetales, 8 (50%) were identified by cytology and culture, 5 (31%) were identified by culture only, and 3 (19%) were identified by cytology only. Of the 129 cases of mycobacteria, 63 (49%) were identified by cytology and culture, 44 (34%) were identified by culture only, and 22 (17%) were identified by cytology only. Among the 67 cases of fungi, 34 (51%) were identified by cytology only, with 15 of these 34 cases being fungal hyphae; 25 cases (37%) were identified by cytology and culture, with a 100% concordance between the cytology diagnosis and culture result; and 8 cases (12%) were identified by culture only. CONCLUSIONS: FNA cytology-culture correlation is a valuable tool with which to assess the efficacy and limitations of the direct diagnosis of infectious agents, and to identify types of infections that may be negative on culture but positive on cytology diagnosis.

Mycobacterium simiae pseudo-outbreak resulting from a contaminated hospital water supply in Houston, Texas.

Various species of nontuberculous mycobacteria are known to cause nosocomial pseudo-outbreaks, but there have been no detailed reports of nosocomial Mycobacterium simiae pseudo-outbreaks. From April 1997 through February 2001, we recovered 65 M. simiae isolates from 62 patients at a community teaching hospital in Houston, Texas. The organism was grown in various water samples obtained in the hospital building and in professional building 1 but not in professional building 2, which has a separate water supply system. Thirty-one environmental and human outbreak-related M. simiae isolates had indistinguishable or closely related patterns on pulsed-field gel electrophoresis and were considered clonal. M. simiae can be a cause of nosocomial pseudo-outbreaks. The reservoir for this pseudo-outbreak was identified as a contaminated hospital water supply.

Mycobacterium simae outbreak associated with a hospital water supply.

OBJECTIVE: Mycobacterium simiae is found primarily in the southwestern United States, Israel, and Cuba, with tap water as its suspected reservoir. Our institution saw an increase in M. simiae isolates in 2001. An investigation into possible contaminated water sources was undertaken. DESIGN: Environmental cultures were performed from water taps in the microbiology laboratory, patient rooms, points in the flow of water to the hospital, and patients' homes. Patient and environmental M. simiae were compared by PFGE. SETTING: Military treatment facility in San Antonio, Texas. PATIENTS: All patients with cultures positive for M. simiae between January 2001 and April 2002. Medical records were reviewed. RESULTS: M. simiae was recovered from water samples from the hospital, patients' home showers, and a well supplying the hospital. A single PFGE clone was predominant among water isolates (9 of 10) and available patient isolates (14 of 19). There was an association between exposure to hospital water and pulmonary samples positive for the clonal M. simiae strain (P = .0018). Only 3 of 22 culture-positive patients met criteria for M. simiae pulmonary disease. Of them, two had indistinguishable M. simiae strains from tap water to which they were routinely exposed. CONCLUSIONS: This represents an outbreak of M. simiae colonization with one nosocomial infection. It is only the second time that M. simiae has been recovered from hospital tap water and its first presentation in municipal water. This study raises issues about the need and feasibility of eliminating or avoiding exposure to M. simiae from tap water.

Anaerobiospirillum succiniciproducens sepsis in an autopsy patient: A troublesome diagnostic workup.

Anaerobiospirillum succiniciproducens is an uncommon yet potentially lethal gram-negative bacterium typically affecting patients with comorbidities. We report a case of A. succiniciproducens infection in an autopsy patient who had hepatitis C and type 2 diabetes and describe the difficulties in the laboratory identification of this pathogen.

Pulsed-field gel electrophoresis study of Mycobacterium abscessus isolates previously affected by DNA degradation.

DNA degradation (which results in a smear pattern) occurs with almost 50% of Mycobacterium abscessus strains during pulsed-field gel electrophoresis (PFGE). We assessed the potential benefit of using thiourea-containing buffer with M. abscessus by studying 69 isolates not previously typeable by PFGE (i.e., those with a smear pattern). Random (epidemiologically unrelated) isolates that were typeable (no smear pattern) were included as controls. Genomic DNA was digested with DraI, XbaI, and AseI. PFGE gels were run in regular gel buffer with and without 100 muM thiourea. All 69 isolates that generated smear patterns had clear band profiles when the thiourea buffer was used. These isolates were divided into only 30 patterns with DraI, 20 patterns with XbaI, and 20 patterns with AseI. The molecular profiles were all closely or possibly related, and the differences between the isolates ranged from zero to six bands. By multilocus enzyme electrophoresis (MEE), 45 of 53 smear isolates (85%) belonged to two closely related electrophoretic types. These isolates contained at least one enzyme allele seen almost exclusively in this group. Isolates without smear patterns were unaffected by thiourea and produced unrelated PFGE profiles, as well as multiple MEE types. The hsp65 and 16S rRNA gene sequences of the isolates with smear patterns were identical to those of M. abscessus type strain ATCC 19977, which had a nonsmear pattern, suggesting that this clone is a subgroup within M. abscessus. This demonstrates that the inability to type M. abscessus by PFGE is associated with a single clone of organisms.

Multisite reproducibility of results obtained by two broth dilution methods for susceptibility testing of Mycobacterium avium complex.

A multicenter study was conducted to assess the interlaboratory reproducibility of susceptibility testing of Mycobacterium avium complex (MAC) by broth microdilution using two different media (cation-adjusted Mueller-Hinton broth with 5% oleic acid-albumin-dextrose-catalase and 7H9 broth with casein) and by macrodilution using the BACTEC 460TB and 12B media at pH 6.8 and 7.3 to 7.4. Ten well-characterized strains of MAC (four macrolide susceptible, six macrolide resistant) were tested against clarithromycin and azithromycin (the latter only by BACTEC 460TB, pH 6.8). At each site, strains were tested against clarithromycin three times on each of three separate days (nine testing events per isolate) by using a common lot of microdilution trays and BACTEC 12B medium, pH 6.8; strains were tested once on three separate days against clarithromycin in 12B medium at pH 7.3 to 7.4 and against azithromycin. Agreement among MICs (i.e., mode +/- 1 twofold dilution) was 100% for all strains and both drugs when BACTEC 460TB was used, regardless of the pH of the medium, but varied when microdilution with either medium was used, particularly with susceptible strains. Agreement based on interpretive category, with NCCLS-recommended breakpoints, was 100% for all strains with the BACTEC 460TB method (both drugs and both pH values) and with microdilution using 7H9 broth. With microdilution and Mueller-Hinton broth, agreement by interpretive category was 100% for eight isolates and >90% for two; errors occurred only in laboratories where personnel had minimal experience with this technique. MAC susceptibility testing may be performed by broth macrodilution or microdilution at either pH, with NCCLS-recommended interpretive breakpoints. However, because visual interpretation of broth microdilution end points is subjective, it is more prone to reader error; therefore, this method requires greater expertise than the BACTEC 460TB. Both techniques require appropriate validation and continued documentation of proficiency.