Bacterial contamination of donated platelets is a major issue in transfusion medicine. Platelet concentrates (PCs) are highly susceptible to bacterial contamination due to their storage conditions at room temperature, which offers a suitable environment for bacterial proliferation. Therefore, the aim of the study is to determine the prevalence of bacterial contamination in the donated platelets units and to identify the isolated microorganisms in samples collected in blood bank services at King Abdullah Medical City (KAMC) in Makkah, Saudi Arabia. A total of 4,318 PCs units including random and pooled platelets were screened for bacterial contamination in the microbiology laboratory. All PCs units were inoculated into blood bottles and loaded into the BacT/ALERT microbial detection system to screen for any suspected bacterial growth over five days. Positive platelet units were subcultured on nutrient agar and incubated overnight at 37°C. The microbiological identification of contaminated PCs units was then performed using the Microscan WalkAway system. Six positive bacterial isolates were recovered and isolated from the screened PCs units. Most of the microorganisms isolated from contaminated platelets were coagulase-negative staphylococci (CoNS), including Staphylococcus capitis (50%), Staphylococcus epidermidis (17%), and Corynebacterium species (33%) among the isolates. Microorganisms isolated from PCs units were mainly skin contaminants related to procedures used during donor phlebotomy or lab processing techniques. However, the rate of bacterial contamination of the donated PCs units subjected to screening was low in this study. The continuous improvement of safety precautions, protocols, and quality assurance of blood collection and storage procedures is essential to reduce the risk of contaminating donated units and any potential post blood transfusion reactions.
Mutations in the pncA gene of Mycobacterium tuberculosis, which encodes the PZase enzyme, are closely linked to pyrazinamide (PZA) resistance. Two clinical isolates, R1 and R2, which are resistant to pyrazinamide at concentrations of 100 μg/mL and 150 μg/mL, respectively, harbor multiple pncA mutations. The pncA R1 gene carries T41C, G419A, and A535G, causing Cys14Arg, Arg140His, and Ser179Gly substitutions in PZase; the pncA R2 gene has G76T, G112C, A403C, and G419A, resulting in Ala26Ser, Ala38Pro, Thr135Pro, and Arg140His changes. The relationship between the biochemical characteristics of the enzymes with pyrazinamide resistance in isolates R1 and R2 remains unclear. This paper presents the catalytic kinetics of mutant enzymes and explores their association with PZA resistance. The impact of these mutations on enzyme function was evaluated by expressing recombinant PZase in Escherichia coli, resulting in the production of proteins approximately 21 kDa in size, as confirmed by SDS-PAGE analysis. Kinetic analysis revealed reduced catalytic efficiency (kcat/KM) for the PZase-R1 and PZase-R2 variants, measured at 1.759 mM¯¹·min¯¹ and 1.500 mM¯¹·min¯¹, respectively, in comparison to the wild-type enzyme, which exhibited a value of 2.443 mM¯¹·min¯¹. These reductions correspond to decreases of 28.0% for PZase-R1 and 38.6% for PZase-R2. The observed declines in catalytic efficiency are likely to contribute to the development of resistance to pyrazinamide (PZA). Further studies, including site-directed mutagenesis and structural modelling, are needed to clarify the impact of each mutation on PZase function and drug interaction.
Current recommendations for screening Chlamydia trachomatis (CT) and Neisseria gonorrhoeae in urinary tract infections using nucleic acid amplification tests (NAATs) include vaginal swabs for women and first‑catch urine for men. In addition, some manufacturers require the use of a transport medium for prolonged pre‑analytical storage. The aim of this study was to evaluate the sensitivity and specificity of midstream urine specimens and to assess nucleic acid stability in neat urine samples stored at +4°C. The study included 338 asymptomatic participants undergoing screening for Chlamydia trachomatis and Neisseria gonorrhoeae as part of the French national STI screening program (172 women and 166 men). For each participant, a reference specimen (self‑collected vaginal swab for women and first‑catch urine for men) was compared with a midstream urine sample. Two comparisons were conducted to assess nucleic acid stability: the first compared urine stored in a preservation medium with neat urine stored at +4°C, and the second compared neat urine stored at +4°C on days 1 and 7. All samples were analysed using the Alinity m Abbott® automated system with the STI AMP Kit. Among the 46 positive samples, the prevalence of Chlamydia trachomatis (CT) and Neisseria gonorrhoeae (NG) was 43/338 and 4/338, respectively. Due to the low prevalence of NG, sensitivity and specificity analyses for midstream urine were limited to CT. Vaginal swabs showed superior performance for detecting CT in females, with two infections missed by midstream urine. Midstream urine demonstrated adequate sensitivity in females (92.9%) and excellent sensitivity in males (100%), with no false-positive results observed. Regarding nucleic acid stability, no statistical difference (p=0.799) was observed for cycle values between urine samples eluted into the liquid medium and neat urine stored at +4°C. Furthermore, our study validated the use of prolonged storage periods (up to 7 days) at +4°C for CT and NG detection in neat urine (p=0.671). In conclusion, the results of this study corroborate the CDC's 2014 recommendations, but the use of midstream urine could offer interesting prospects for Chlamydia trachomatis screening in specific populations. Regarding Neisseria gonorrhoeae, given the low prevalence of positive results found in our study, no meaningful conclusions can be drawn.
This study aims to evaluate the distribution and antimicrobial resistance profiles of microorganisms isolated from blood cultures of patients hospitalised in intensive care units and wards of our hospital over a 2-year period, in light of European antimicrobial resistance data and country surveillance data. Blood cultures sent to the microbiology laboratory from the wards and intensive care units of our hospital between December 2020 and 2022 were retrospectively evaluated from the hospital automation system. The identification and the antimicrobial susceptibility of the isolates were done by the Phoenix 100 system (Becton Dickinson, USA). The results of antibiotic susceptibility were evaluated according to the criteria of the European Committee for Antimicrobial Susceptibility Testing (EUCAST). The blood culture results of 872 patients with pathogenic bacterial growth in their blood cultures were examined and evaluated. Taking into account World Health Organisation (WHO) surveillance reports, it appears that our hospital's antimicrobial resistance rates were higher than those reported by our country's data. Resistance profiles vary from country to country, city to city, and even among different hospitals in the same city. Monitoring of antimicrobial resistance, even at the hospital level, will contribute to programs to combat antimicrobial resistance both across the country and all around the world.
We present a comparison of the mucosal microbiota within different diverticula in a patient with diverticular disease (DD) complicated by diverticulitis and pelvic abscess. The conventional culture method and the 16S rRNA-based sequencing approach were employed to characterize the microbiota of perforated diverticulum (PD) and adjacent visually intact diverticulum (ID) from the same surgically resected colonic segment. Compared to PD, the microbiota of ID demonstrated depletion in butyrate-producing genera and increased abundances of Proteobacteria, Enterobacteriaceae, and Bacteroides. The predominantly pro-inflammatory character of the microbiota in ID suggests its probable pathological role in the progression of DD towards more complicated forms, up to inflammatory destruction (perforation) of the diverticulum wall. The insights of this study pave the way for the development of forthcoming clinical trials focusing on microbiota-related therapies, including the use of antibiotics, probiotics, and fecal microbiota transplantation (FMT), to potentially treat or manage DD and its complications.
Malaria remains a significant global health challenge, disproportionately affecting Sub-Saharan Africa. Pregnant women represent one of the most vulnerable populations. Despite numerous advances in malaria control measures, lost-to-follow-up (LTFU) in antenatal care (ANC) programs poses a critical barrier to achieving optimal maternal and neonatal outcomes and the success of public health interventions. The factors driving LTFU, particularly in rural, high-burden settings, are not completely understood. This study investigates the determinants of LTFU among pregnant women receiving malaria screening in the context of the ERASE-Rise Against Malaria project in Northern Uganda. An observational retrospective cohort study was conducted on 1,558 women, recruited from July 2022 to June 2024, during the operational research held to assess the impact of antimalarial resistance on malaria care among pregnant women in three healthcare facilities in Oyam and Kole districts. Data on individual level (sociodemographic and clinical) and health care-related factors were analyzed using a multilevel logistic regression model to identify predictors of LTFU, defined as the absence of cohort outcome data 30 days after the expected delivery date. Efforts to recover missing data included delivery register consultation, active phone calls, and contact tracing by village health team workers. 871 (55.9%) of the 1,558 women were LTFU. Recovery strategies reduced the missing data rate to 29.1% (n=454). Protective factors against LTFU included higher education (aOR=0.75, 95% CI: 0.54-1.03, p=0.0798), being primigravida (aOR=0.73, 95% CI: 0.56-0.97, p=0.0275), and experiencing malaria during pregnancy (aOR=0.61, 95% CI: 0.48-0.78, p<0.0001). Women attending Aboke Health Center IV (n=385) were over five times more likely to be LTFU than those at Aber Hospital (n=955) (aOR=5.57, 95% CI: 4.08-7.71, p<0.0001), highlighting significant geographic and structural barriers. The high rate of LTFU in malaria screening programs underscores the need for targeted interventions addressing individual, systemic and structural barriers. Strengthening community-level support, improving healthcare infrastructures, and integrating malaria prevention into broader maternal health services are crucial for enhancing retention in care. Addressing determinants of LTFU, systematically, through further qualitative and quantitative research, is essential to improving maternal and neonatal health outcomes and achieving malaria eradication goals in high-burden settings.
Paranasal sinus Fungus Ball (FB) is the most common non-invasive mycotic rhinosinusitis. It most frequently affects the maxillary and sphenoidal sinuses and the treatment of choice is Endoscopic Sinus Surgery (ESS). Although this pathology has been widely investigated throughout the years, some questions still remain unanswered. This study concentrates on assessing radiological and microbiological characteristics by examining a large number of cases treated in our centre. 235 cases of FB that underwent ESS in Fondazione I.R.C.C.S. Policlinico San Matteo di Pavia in the period comprised between January 2000 and May 2020 were collected. The surgical report, microbiological culture, histological report and preoperative Computed Tomography were analysed. FB was confirmed to affect more commonly the female population (68.22%). The maxillary (69.78%) and sphenoidal sinuses (27.23%) were the most frequent localizations. Interestingly, the microbiological reports showed different growth patterns as positive cultures from maxillary FB were reached in 21.52% of cases, while from sphenoidal FB in 45.76%. Different mycotic populations were found: Aspergillus fumigatus was isolated in 33 FB specimens, of which 63.3% in the maxillary sinus, while Aspergillus flavus was isolated in 10 specimens, of which 80% in the sphenoid sinus (p=0.017 and p=0.039 respectively). Radiologically, heterogeneous soft tissue density at sinus cavity (p=0.029) was more represented in patients with positive culture. This study analysed a large population and demonstrated differences in the growth pattern and subpopulation of fungi between differently localized FB, underlining a new characteristic of this pathology.
Acute respiratory infections (ARIs) are among the most common infections, with variable etiology, affecting both community and hospital settings. Vulnerable populations include children, the elderly, and patients with chronic debilitating disease. Early identification of the causative pathogen can reduce inappropriate empirical antibiotic therapy, particularly in viral infection cases, which are the leading cause of ARIs, especially in outpatient settings. Rapid molecular diagnostic methods such as FilmArray Respiratory Panel (FA RP) enable the prompt identification of viral and bacterial pathogens. A retrospective observational study was conducted at Santa Maria Goretti Hospital in Latina, Italy, from January 1, 2022, to April 30, 2023. A total of 232 nasopharyngeal swabs from patients were analyzed using the BioFire FA RP. Eligible patients exhibited fever, cough, or other symptoms suggestive of respiratory infection. The test distribution shows that the Pediatric and Neonatology Units had the highest number of tests and a high proportion of clinically relevant outcomes. The findings underscore the effectiveness of the FA RP in swiftly identifying pathogens, significantly reducing diagnostic turnaround times, and minimizing unnecessary antibiotic prescriptions. The 38% rate of negative tests highlights the importance of appropriate test prescription based on clinical assessment.
Candiduria is a common problem especially in advanced stage oncology/hematology cases. Herein, we aimed to analyze the efficacy of caspofungin treatment in Candida auris isolated from urine culture. We conducted an observational retrospective study in a tertiary-care educational hemato-oncology hospital from October 2021 to November 2022. The patients hospitalized in our center and having at least two consecutive urine cultures that yielded C. auris with significant pyuria were included in the study. The effectiveness of caspofungin was evaluated based on clinical and microbiological outcomes. In total, 10 patients (two female, aged 55.3 ± 15) were included in the study. Four (40%) patients had urinary tract infections, and six (60%) patients had asymptomatic C. auris candiduria which required antifungal therapy. Nine cases had urinary instrumentation: two nephrostomy, one suprapubic catheter, and six urinary catheters. The 10 cases received a mean of 13.4 ± 3.5 days of caspofungin. Four of 10 cases (40%) had at least 1 negative culture between day 1 and the end of caspofungin treatment. Relapse and re-infection rates 30 days after the end of therapy in microbiologically successful cases were 3/4 and 2/4. Caspofungin resistance developed in three cases (30%) within 30 days after treatment. Overall day-30 mortality was 3/10 (30%) while overall success (microbiological success + no relapse/reinfection during 30 days follow up after end of therapy) was 10%. Caspofungin demonstrated limited efficacy in treating C. auris-related urinary tract infection/colonization in hospitalized cancer patients.
To understand the resistance phenotype and the carriage of resistance genes in carbapenem-resistant Serratia marcescens in our hospital. Two S. marcescens clinical isolates were analyzed retrospectively to determine their sensitivity to 27 antimicrobial agents using the BD Phoenix™ M50 System (Becton, Dickinson and Company, Franklin Lakes, NJ, USA) and Kirby-Bauer disk diffusion method (Oxoid, Hampshire, UK). Whole-genome sequencing of both strains was performed using the Illumina platform. ABRicate software was used to predict the resistance genes and plasmid replicon types carried by the strains. Sequence type (ST) analysis was performed using multilocus sequence typing. A phylogenetic tree was constructed, and homology analysis was conducted by comparing the two strains with 106 blaKPC-2-carrying carbapenem-resistant S. marcescens isolates downloaded from the NCBI database. Two S. marcescens strains were isolated from the sputum samples of patients with lower respiratory tract infections after ICU surgery. Antimicrobial sensitivity testing revealed that both strains were resistant to antimicrobial agents, including β-lactams, sulfonamides, and carbapenems, whereas they were susceptible to tigecycline and ceftazidime/avibactam. In total, 12 resistance genes were predicted, with both WF0070 and WF0071 carrying the carbapenem resistance gene blaKPC-2. Among the remaining resistance genes, the aminoglycoside resistance genes were most commonly predicted, with five types identified. The two strains in this study and the 106 blaKPC-2-carrying carbapenem-resistant S. marcescens isolates downloaded from NCBI were divided into five evolutionary groups, and both strains in this experiment were categorized into clade E. Meanwhile, 18 STs were predicted, and both strains in this study belonged to ST366. In terms of phylogenetic relationships, these strains exhibited high homology with a strain isolated in France in 2018. The two S. marcescens strains in this study displayed high resistance to multiple antimicrobial agents, and both carried the carbapenem resistance gene blaKPC-2 along with several other resistance genes. The two strains displayed close phylogenetic relationships, and the possibility of nosocomial transmission cannot be dismissed.
In European countries, the number of locally-acquired hepatitis E virus (HEV) cases has increased in recent years, primarily linked to infections caused by HEV genotype 3 (HEV-3). The novel HEV-3 subtype 3n was initially reported in Italy; however, its distribution across other European regions remained uncertain. Here, we analyzed all HEV-3 ORF2 sequences from European countries deposited in NCBI (2016-2024). Phylogenetic analysis identified 47 HEV-3n sequences: 43 from wild boars (37 from Italy, 6 from Poland) and 4 human-derived sequences (two blood donor samples each from France and Denmark). Nucleotide sequence analysis demonstrated a 4.0-7.2% genetic distance between human and wild boar HEV-3n isolates, confirming their close evolutionary relationship. This genetic evidence, combined with the geographic distribution across Italy, Poland, France, and Denmark, provides robust molecular support for zoonotic transmission of HEV-3n from wild boars to humans. The findings indicate that HEV-3n is not restricted to Italy but has disseminated across multiple European countries, establishing wild boars as a key reservoir for human infections. This study underscores the need for integrated surveillance of HEV-3n in wildlife and human populations across Europe to mitigate zoonotic risks.
Rapid and accurate testing of colistin resistance is of paramount importance for the effective management of gram-negative infections in workflow laboratories. The objective of this study was to evaluate the performance of commercial VITEK®2 AST-XN21 cards and ChromID media via the reference Diagnostics MIC-COL method. Colistin susceptibility testing was conducted on 72 nonduplicate isolates using Diagnostics MIC-COL as the reference method, in comparison with the VITEK®2 AST-XN21 card and ChromID agar. The colistin resistance rate was 37.5% (27/72) according to the Diagnostics MIC-COL (Klebsiella spp. 34.4% (n=10), and Acinetobacter baumannii (A. baumannii) 39.5% (n=17)), whereas the VITEK®2 AST-XN21 card resistance rate was 41.3% (n=12) for Klebsiella spp. and 48.8% (n=21) for A. baumannii. The VITEK®2 AST-XN21 card was CA 91.6%, and the very major error (VME) and major error (ME) rates for all the isolates were 1.38% (n=1) and 6.9% (n=5), respectively. A. baumannii isolates were CA (37/43) 86%, VME (1/43) 2.3% and ME (5/43) 11.6% with XN21 cards. For Klebsiella spp., CA (29/29) was 100%, whereas ME and VME were not detected. The CA was 94.4% (68/72) for ChromID medium, 93% (40/43) for A. baumannii and 96.5% (28/29) for Klebsiella spp. These results indicated that the CHROMID medium is an appropriate method for the screening of colistin resistance. Nevertheless, it is imperative to exercise caution when interpreting the results of a commercial automated VITEK®2 AST-N21 card for colistin susceptibility testing.
Infections caused by multidrug-resistant Gram-negative bacteria, particularly carbapenemase-producing Enterobacterales, present significant diagnostic and therapeutic challenges, highlighting the need for practical laboratory methods to evaluate combination therapies. This study investigated 90 ceftazidime-avibactam (CZA)-resistant Klebsiella pneumoniae isolates collected from clinical specimens between January 2021 and July 2023. Species identification was performed using MALDI-TOF MS, and antimicrobial susceptibility testing was conducted with the BD Phoenix™ system according to Clinical and Laboratory Standards Institute (CLSI) criteria, with disk diffusion used for confirmation when necessary. Carbapenemase production was determined phenotypically using the modified Carbapenem Inactivation Method. For a subset of isolates with available whole-genome sequencing data, resistance genes were confirmed, although genotypic analysis was not performed for all isolates. The in vitro activity of the CZA plus aztreonam (CZA-ATM) combination was evaluated using the CLSI-recommended broth disk elution method. All isolates were resistant to CZA and aztreonam when tested individually, whereas no visible growth was observed with the combination, indicating in vitro susceptibility. These findings suggest that the broth disk elution method is a practical approach for assessing CZA-ATM activity in routine clinical laboratories. Further studies including reference method validation and evaluation of borderline or resistant isolates are needed.
Yersinia enterocolitica is a food-borne bacterium related to a disease called yersiniosis (gastroenteritis) in animals and humans. It is widely distributed in nature and is controlled with various antibiotics, such as second- and third-generation cephalosporins, gentamicin and fluoroquinolones and ciprofloxacin. In addition to antibiotic resistance, these antibiotics have side effects in the host because of drug-host interaction. To overcome these problems, this study focused on finding putative drug targets in the pathogen Y. enterocolitica and identifying novel molecules to tackle the function of this pathogen. In the present study, two new drug targets were found in Y. enterocolitica. Apart from using antibiotics, chlorogenic acid, hesperidin, neohesperidin, and naringin had indicated good binding affinity with the recognized targets through molecular docking experiments. Accordingly, this investigation suggests that the newly-found drug targets may be a fertile field in the pharmaceutical community for finding inhibitors similar to chlorogenic acid, hesperidin, neohesperidin, and naringin, as well as designing new drug molecule formulations to tackle the foodborne illness caused by pathogen Y. enterocolitica.
The introduction of integrase strand transfer inhibitors (INSTIs) has transformed HIV therapy, offering high efficacy and tolerability. However, emerging evidence links INSTI exposure to weight gain. The long-term impact of switching from protease inhibitor (PI)-based to INSTI-based combined antiretroviral therapy (cART) on body composition and metabolic health remains incompletely understood. We conducted a retrospective longitudinal study of 89 virologically suppressed people living with HIV (PLWH) followed from 2008 to 2021. All participants were on PI-based cART at baseline (T0). Between 2008 and 2013 (T1), a subset switched to a raltegravir-based INSTI regimen, with subsequent transitions to dolutegravir or bictegravir during follow-up (T2). Changes in body weight, body mass index (BMI), and metabolic parameters were compared between participants who remained on PI therapy and those who switched to any INSTI ("Ever INSTI"). Over the full observation period, individuals who switched to INSTI-based therapy experienced significantly greater mean increases in body weight and BMI compared with those maintained on PI-based regimens. Immune recovery remained stable across groups, and metabolic safety appeared preserved. Within the INSTI class, participants with prolonged raltegravir exposure exhibited a trend toward greater long-term weight gain, though this finding should be interpreted cautiously due to the small subgroup size and potential residual confounding. Switching from PI- to INSTI-based cART in virologically suppressed PLWH is associated with modest but sustained increases in body weight and BMI over time, without evident deterioration in metabolic health. These results support the favourable safety profile of INSTIbased regimens while underscoring the need for routine anthropometric monitoring and preventive lifestyle interventions during long-term therapy.
Staphylococcus aureus, commonly colonizing the mucous membranes and skin of humans, is a prevalent pathogen responsible for Skin and Soft Tissue Infections (SSTIs), with a notably high prevalence of methicillin-resistant Staphylococcus aureus (MRSA). The antimicrobial resistance profiles of MRSA vary across different regions, with differences in population structure and epidemiological characteristics over time and geography. Molecular typing is frequently employed to investigate the population structure and transmission features among strains. The purpose of this study is to investigate the molecular epidemiological characteristics, virulence genes, and drug resistance of MRSA isolated from patients with skin and soft tissue infections in the Shaoxing region. Seventy-seven MRSA strains isolated from patients with SSTIs in the Shaoxing area from 2017 to 2022 were subjected to drug sensitivity testing using the VITEK 2 Compact fully automated system. The study utilized next-generation sequencing to conduct Staphylococcal Chromosome Cassette mec (SCCmec) typing, Staphylococcal Protein A (SPA) typing, Multilocus Sequence Typing (MLST), and investigate virulence genes in MRSA strains. The variations in antimicrobial resistance, virulence genes, and molecular typing among different genders and age groups were analyzed. The findings revealed that the resistance rate of MRSA to penicillin was 100%, while it was 61.04% for erythromycin and 59.74% for clindamycin. The resistance rate to other antibiotics was below 20%. MLST typing is mainly dominated by ST59 (22.08%) and ST398 (18.18%), while SCCmec typing is predominantly represented by IV (38 strains) and V (27 strains). SPA typing is mainly characterized by t437 (19.48%) and t34 (14.29%). The major clones of SSTIs in the Shaoxing area are ST59-t437-IV and ST398-t34-V. The strains carry 17 enterotoxin genes, with the highest detection rates found in sek and seq (32.47%), PVL (12.99%), and tst (7.79%).This study demonstrates that skin and soft tissue infections in the Shaoxing region are caused mainly by ST59-t437-IV and ST398-t34-V strains, which carry multiple virulence genes. PVL is identified as a significant virulence factor in MRSA strains.
Methicillin-resistant Staphylococcus aureus has become significant due to its prevalence as one of the main causes of nosocomial infections, provoking both human and economic losses; therefore, a better understanding of its biology will allow the proposal of new strategies of control. To analyze the metabolic profiles of two strains of Staphylococcus aureus under exposure to sub-inhibitory concentrations of oxacillin. Methicillin-susceptible Staphylococcus aureus and Methicillin-resistant Staphylococcus aureus strains were exposed to oxacillin at 0.125 mg/L; afterwards, tandem mass spectrometry was used to analyze their metabolic profiles (12 amino acid [AA]s and 28 acylcarnitine [AcC]s). Exposure to oxacillin in both strains generated osmolytes such as proline and carnitine, in response to osmotic stress generated from the damage on the cellular wall. Moreover, they presented a capacity to modify their intra-cellular composition of both amino acids and AcCs in response to exposure to the drug. Exposure of MSSA and MRSA strains to oxacillin modifies their metabolome.
We present the case of a 47-year-old man living with HIV who had experienced fever, night sweats, cough and occasional haemoptysis for the previous two months. Computed tomography of the chest revealed a lesion measuring 57 mm in diameter with cavitation in the left lower superior segment. His CD4+ T cell count was 220 cells/mm³ and his HIV RNA level was 406,756 copies/ml. Acid-resistant coccobacilli were isolated from the transthoracic biopsy of the lesion and were initially identified as Mycobacterium lentiflavum. However, 16S rDNA sequence analysis identified it as Rhodococcus equi. A histopathological examination revealing malakoplakia with Michaelis-Gutmann bodies supported the diagnosis. The patient responded dramatically to a 3-month course of treatment with azithromycin and moxifloxacin. The most obvious epidemiological risk factor in this case was that the patient regularly took care of multiple stray cats. Rhodococcus equi should be considered in differential diagnosis of cavitary pneumonia in patients with cellular immunodeficiencies, especially when there is a contact history not only with horses, but with other herbivores.
In this narrative review, inspired by the observation of changes in clinical practice implemented at our centre (IRCCS San Gerardo dei Tintori), we describe the evolution of care for pregnant women living with HIV (WLWH) and the current issues regarding ARV therapy, delivery, prophylaxis and breastfeeding. Over the years, advances in the care of HIV in pregnancy have significantly reduced the risk of perinatal transmission, making the experience of motherhood in WLWH comparable to that of people without HIV infection. However, some issues remain to be overcome to separate the real risks of transmission from the perceived fears and reduce the medicalization of pregnancy in WLWH. Notably, the rate of caesarean section remains higher than that of the general population, highlighting the need to further promote vaginal birth for WLWH in clinical practice and to reassure both women and physicians on its safety. Moreover, certain areas remain uncertain or subject to conflicting guidelines, such as the use of post-exposure prophylaxis for low-risk neonates and breastfeeding.
Dermatophytes are the leading cause of cutaneous fungal infection worldwide, followed by non-dermatophyte molds and Candida spp. Microbiological diagnosis is essential to identify the etiological agent and to guide targeted therapy. Conventional methods, including direct microscopy and fungal culture, are fundamental tools in the diagnostic workflow. This study aimed to assess quantitative Multiplex Tandem PCR performance in comparison with microscopy and culture for rapid and accurate identification in real clinical practice. A total of 50 clinical specimens were collected and analyzed with conventional methods. Direct microscopic examination yielded positive results in 45 out of 50 samples (90%), while fungal culture identified clinically significant fungi in 29 out of 50 cases (58%). The same 50 specimens were retrospectively analyzed using Multiplex MT-PCR, which detects major dermatophytes, Scopulariopsis, Aspergillus and Candida species. MT-PCR identified fungal DNA in 84% of samples. Compared to standard reference methods, the assay demonstrated a sensitivity of 84% and a specificity of 60%. Notably, MT-PCR identified fungal pathogens that were missed by conventional techniques in 32% of cases, significantly reducing the turnaround time. Conversely, 6% of samples were PCR-negative but culture-positive, likely reflecting pre-analytical variability and sample heterogeneity. Although the sample size was limited, MT-PCR proved to be a rapid and highly sensitive complementary diagnostic tool, warranting further investigation in larger-scale studies.