Standard Culture Methods Research Articles

Improving neonatal outcome by studying the incidence of maternal group B streptococcus colonization among pregnant women: A prospective observational study

ABSTRACT Aims and Objectives: Group B streptococcus (GBS) colonization during pregnancy can lead to invasive infections in neonates including meningitis or sepsis. The major risk factor for neonatal disease is the rectovaginal colonization of GBS during pregnancy and delivery. The objectives of this study were as follows: 1) to screen antenatal patients at 35–37 weeks of gestation for GBS colonization, 2) to determine the prevalence of GBS among pregnant women, 3) to formulate an antibiotic policy with respect to antepartum antibiotic prophylaxis and empiric choice of antibiotics for early onset neonatal sepsis, and 4) to ascertain the maternal and neonatal outcomes in pregnant women with positive GBS colonization. Material and Methods: A prospective observational study was conducted among 50 obstetric patients at 35–37 weeks of gestation attending the outpatient and inpatient departments of a tertiary care hospital. High vaginal and rectal swabs were collected and immediately transported to the microbiology lab and were sent for culture and sensitivity testing using the standard culture methods. The incidence of positive cultures was noted and was taken up as a reference for statistical calculations. Results: Out of the 50 vaginal samples, one sample showed GBS positivity having a prevalence of 2%. Rectal swabs did not grow GBS for all samples. Neither the pregnant woman nor the newborn showed any symptoms or signs due to GBS. GBS grown in vaginal culture was highly sensitive to benzyl penicillin and other penicillins, cephalosporins, clindamycin, daptomycin, vancomycin, meropenem, and linezolid. Conclusion: Based on our findings, screening the patients with routine rectovaginal swabs may not be necessary in our population. Also, empiric treatment with antibiotics for GBS in symptomatic mothers cannot be justified, and hence, GBS should be considered as an insignificant pathogen.

Cockroaches as Reservoirs, Vectors, and Potential Sentinels of Multidrug-Resistant Bacteria in Ugandan Communities: A Retrospective Analysis.

Background: Cockroaches could play a role in the transmission dynamics of antimicrobial-resistant bacteria (ARB) at variable interfaces in Ugandan communities, acting as both reservoirs and vectors. This study investigated the burden and diversity of ARB carried by cockroaches in human settlements in Uganda, so as to understand their role in the spread of these pathogens and their potential as sentinels in antimicrobial resistance (AMR) surveillance programs. Materials and Methods: A retrospective analysis was conducted on two unpublished studies by Makerere University students. Study one and study two sampled 58 and 110 cockroaches, respectively, from secondary schools in Kampala. Cockroach species identification was determined based on physical characteristics. Bacterial isolation and characterization were performed through microbiological analyses including standard culture methods, biochemical tests, matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS), disc diffusion method, and polymerase chain reaction (PCR). Results: Majority of the cockroaches (over 80%) were Periplaneta americana. Multidrug resistance (MDR) was prevalent among the isolates, with over 30% of the isolates being resistant to three or more antibiotic classes. Specifically, MDR (over 90%) was rampant in the extended spectrum beta-lactamase (ESBL)- or AmpC-producing Escherichia coli and enterococci isolates. Critical World Health Organization (WHO) priority pathogens, such as ESBL-/AmpC-producing Enterobacteriaceae, and carbapenem-resistant E. coli, were also identified. The most abundant resistance determinants (tetracycline and sulphonamide) were tetA, sul1, and sul2 for E. coli, and tetM and tetL for enterococci. Conclusion: The findings accentuate the potential role of cockroaches: (1) in transmitting multidrug-resistant bacteria at the human-animal-environment interface and (2) as sentinels in the surveillance of community-generated AMR.

Rapid and accurate testing for urinary tract infection: new clothes for the emperor.

SUMMARYUrinary tract infection (UTI) is among the most common infections in clinical practice. In some cases, if left untreated, it can lead to pyelonephritis and urosepsis. In other cases, UTI resolves without treatment. Clinical diagnosis is typically based on patient symptoms and/or urinalysis, including urine dipsticks. The standard urine culture method is sometimes employed to identify the suspected urinary pathogen (uropathogen) and/or guide antimicrobial choice, but results are rarely available before 24 h. The standard urine culture method also misses fastidious, anaerobic, and slow-growing uropathogens and rarely reports polymicrobial infections. The unexplained combination of negative urine cultures with persistent urinary tract symptoms is distressing to both patients and clinicians. Given the broad appreciation of the advantages provided by rapid testing (e.g., for COVID-19 or influenza A), a rapid, accurate diagnostic test is needed to deliver timely treatment to patients seeking care for UTI that optimizes antibiotic stewardship. Herein, we discuss progress being made toward an accessible, timely (i.e., within hours), accurate assay with results that are clinically useful for the treating clinician within the timeframe of the infection (i.e., the growth rate of the pathogen(s)). New and emerging uropathogens often overlooked by current diagnostic techniques are also reviewed.

The diagnostic efficacy of rapid urine sediment fluorescence staining in urinary tract infections: An exploratory study

ObjectiveTo investigate rapid and early diagnostic approaches for urinary tract infections (UTIs). MethodsCoincidence rates, sensitivities, and specificities of fungal fluorescence staining for fungal detection were calculated. Additionally, the positive rate of cellular fluorescence staining was compared with bacterial urine culture, with a focus on the increasing correlation between fluorescently stained bacteria proportions and the concordance between methods. Furthermore, diagnostic performance for detecting cocci and bacilli via histocyte fluorescence staining was evaluated. Receiver Operating Characteristic (ROC) curve analyses were conducted to determine the diagnostic accuracy of urine sediment fluorescence staining for fungal and bacterial UTIs. ResultsFungal fluorescence staining of urine sediment demonstrated a coincidence rate of 95.90%, sensitivity of 94.44%, and specificity of 96.05%. The positivity of cellular fluorescence staining exceeded that of bacterial culture (P=0.014), and an enhanced agreement between the two methods was observed with a rise in the proportion of stained bacteria. For histocyte fluorescence staining in identifying cocci and bacilli, coincidence rates were 76.92% and 74.36%, sensitivities were 51.85% and 73.85%, and specificities reached 82.43% and 88.18%, respectively. The ROC curve areas for diagnosing fungal and bacterial UTIs using urine sediment fluorescence staining were 0.952 and 0.87, respectively. ConclusionUrine sediment fluorescence staining offers promise as a rapid diagnostic tool for UTIs, demonstrating high sensitivity and specificity for fungal detection and a competitive performance in bacterial identification when compared to standard culture methods. These findings highlight the potential utility of this technique in clinical settings for prompt and accurate UTI diagnosis.

Physicochemical and microbiological evaluation of yoghurt sold in Addis Ababa, Ethiopia

BackgroundIncrease in demand for yoghurt in Addis Ababa, in recent times has led to proliferation of different kinds of commercially produced (branded) yoghurt in line with the traditional (unbranded) one in the market. However, limited or no available data for the quality of gradually increasing different kinds of commercially produced (branded) yoghurt in line with the traditional (unbranded) one in the market. It was therefore vital to evaluate the physicochemical property and microbial quality of yoghurt sold in Addis Ababa in order to determine its quality and perhaps safeguard the health and wellbeing of the numerous people consuming yoghurt products.MethodsA total of 40 yoghurt samples consisted of 20 traditionally (unbranded) and 20 commercially produced (branded) were analysis for physicochemical property (pH, moisture, ash, fat, Total Solid (TS) and Solid Not Fat (SNF)) and microbial quality (Coliform Count (CC) and Yeast and Mould Count (YMC) were performed using TEMPO system whereas; Total Viable Count (TVC) was performed according to standard culture method).ResultsIn the traditionally produced (unbranded) yogurt samples, the fat content was 4.44%, pH 3.99, TS content 10.12%, SNF content 7.18%, moisture content 89.88%, and ash content 0.53%. For the commercially produced (branded) yogurt samples, the fat content was 5.02%, pH 3.88, TS content 10.66%, SNF content 8.10%, moisture content 89.29%, and ash content 0.62%. The TVC in traditionally (unbranded) and commercially produced (branded) yoghurt samples was found to be 10.72 and 10.35 log10 cfu/mL, respectively. In terms of coliform counts (CC), 20%, 55%, and 25% of the traditionally produced yogurt samples had counts of < 10 cfu/mL, 10 to < 4.9 × 104 cfu/mL, and > 4.9 × 104 cfu/mL, respectively. For the commercially produced yogurt samples, 40%, 35%, and 25% fell within these same ranges. The overall mean coliform counts were 3.72 log₁₀ cfu/mL for traditionally produced yogurt samples and 2.81 log₁₀ cfu/mL for commercially produced yogurt samples; however, the difference was not statistically significant (P > 0.05). Similarly, for yeast and mold counts (YMC), 0%, 35%, and 65% of traditionally produced yogurt samples had counts of < 100 cfu/mL, 100—< 4.9 × 104 cfu/mL, and > 4.9 × 104 cfu/mL, respectively. In commercially produced yogurt, 30%, 50%, and 20% of samples fell within these same ranges. The overall mean YMC was 4.48 log₁₀ cfu/mL for traditionally produced yogurt samples and 3.92 log₁₀ cfu/mL for commercially produced yogurt samples, but this difference was also not statistically significant (P > 0.05).ConclusionBased on the findings of this study, it is crucial to implement measures to improve the quality of yogurt in Addis Ababa to ensure consumer safety and product consistency.

Adipo-on-chip: a microphysiological system to culture human mesenchymal stem cells with improved adipogenic differentiation.

Obesity is associated with several comorbidities that cause high mortality rates worldwide. Thus, the study of adipose tissue (AT) has become a target of high interest because of its crucial contribution to many metabolic diseases and metabolizing potential. However, many AT-related physiological, pathophysiological, and toxicological mechanisms in humans are still poorly understood, mainly due to the use of non-human animal models. Organ-on-chip (OoC) platform is a promising alternative to animal models. However, the use of adipose-derived human mesenchymal stem cells (hASCs) in these models is still scarce, and more knowledge on the effects properties of culturing hASCs in OoC models is needed. Here, we present the development of an OoC using hASCs to assess adipogenic differentiation. The device capability to increase hASC differentiation levels was confirmed by Nile red staining to verify lipid droplets inside cells after 10days of culture and fluid flow of 10 µL/h. The Adipo-on-a-chip system increases hASC proliferation and differentiation area compared with the standard culture method under static conditions (96-well plates) verified in hASCs from different donors by image analysis of cells stained with Nile red. The expression of the gene FABP4 is lower in the MPS, which calls attention to different homeostasis and control of lipids in cells in the MPS, compared with the plates. An increase of hASC proliferation in the MPS related to the 96-well plate was verified through protein Ki-67 expression. Cell and nuclei morphology (area, roundness, perimeter, width, length, width to length rate, symmetry, compactness, axial and radial properties to nuclei, and texture) and dominant direction of cells inside the MPS were evaluated to characterize hASCs in the present model. The presented microphysiological system (MPS) provides a promising tool for applications in mechanistic research aiming to investigate adipogenesis in AT and toxicological assessment based on the hASC differentiation potential.

Role of Xpert PCR kit in assessing MRSA colonization in medical and surgical units of a tertiary care teaching hospital

Background and rationaleMethicillin resistant Staphylococcus aureus (MRSA) colonization increases the risk of MRSA infection. Detecting MRSA colonization can influence postoperative outcomes and prolong hospital stay. The conventional standard culture method for detecting MRSA colonization has limitations in terms of sensitivity and turnaround time. Hence, we sought out use of Xpert PCR kit for prompt evaluation of MRSA colonization to support MRSA prevention in a tertiary care hospital in Karachi, Pakistan.Materials and methodsDuring 1st April-31st December 2022, 290 nasal and skin swab samples were collected from 257 patients and processed using routine culture (as gold standard method) and PCR-based MRSA detection assay (MRSA Xpert).ResultsA total of two hundred and ninety (290) swab samples from 257 patients were obtained, 33 of which were paired. The overall prevalence of MRSA colonization was 12% by both methods, with 90% of cases classified as community-associated (CA-MRSA) whereas 10% as hospital-acquired (HA-MRSA). The colonized group showed a higher subsequent MRSA infection rate (11% vs. 3.5%) compared to the noncolonized group. Culture identified 11% of screening samples as MRSA positive, Xpert MRSA assay showed 100% sensitivity and 95% specificity. The cost of a single MRSA Xpert assay was $50 while MRSA culture cost around $7.50.ConclusionOur study findings suggest that the presence of MRSA colonization in our cohort of patients is consistent with the existing trends in hospital epidemiology. Both conventional culture and Xpert MRSA methods showed comparable efficacy for detection of MRSA colonization. Larger-scale studies are recommended to validate these findings conclusively.

Urinary tract infection among people living with human immunodeficiency virus attending selected hospitals in Addis Ababa and Adama, central Ethiopia.

Urinary tract infections (UTIs) and antibacterial resistance (ABR) are important public health problems, but they are not well-studied among people living with human immunodeficiency virus (PLHIV) globally, especially in low-income countries. Therefore, it is important to regularly measure the extent of UTIs and ABR in the most susceptible populations. This study aimed to investigate the prevalence of UTIs, associated factors, bacterial causal agents, and their antibiotic susceptibility profile among PLHIV in central Ethiopia. A hospital-based cross-sectional study was conducted to recruit 688 PLHIV by a simple random sampling method. Background information was gathered through interviews, while clinical information was gathered from recent information sheets of patient charts using organized, pretested, and validated study tools. Midstream urine was collected aseptically and transported to the Microbiology Laboratory of Aklilu Lemma Institute of Pathobiology within 4 h of collection, maintaining its cold chain. Standard conventional microbial culture methods and matrix-assisted laser desorption ionization-time of flight (MALDI-TOF) mass spectrometry were used to identify the bacterial isolates at the species level. Kirby Bauer's disk diffusion method was used to determine the antibiotic susceptibility profile of the bacterial isolates based on the interpretation guidelines of the Clinical Laboratory Standard Institute. Logistic regression models were used to examine factors associated with the occurrence of UTIs among PLHIV attending selected hospitals in Addis Ababa, and Adama. Out of 688 PLHIVs involved in the current study, 144 (20.9%) were positive for UTIs, whereas the majority were asymptomatic for UTIs. In the multivariable logistic regression analysis, only HIV RNA ≥ 200 copies/ml [AOR = 12.24 (95% CI, 3.24, 46.20), p < 0.01] and being symptomatic for UTIs during the study period [AOR = 11.57 (95% CI, 5.83, 22.97), p < 0.01] were associated with the occurrence of UTIs. The dominant bacterial species isolated were Escherichia coli (E. coli; n = 65; 43%), followed by Enterococcus faecalis (E. faecalis; n = 16; 10.6%) and Klebsiella pneumoniae (K. pneumoniae; n = 11; 7.3%). Over half of the E. coli isolates were resistant to antibiotics such as gentamicin (GM; n = 44; 67.7%), amikacin (AN; n = 46; 70.8%), nalidixic acid (NA; n = 42; 64.6%), ciprofloxacin (CIP; n = 40; 61.5%), and azithromycin (AZM; n = 45; 69.2%). All of the K. pneumoniae isolates (n = 11; 100%), (n = 6; 54.5%), and (n = 7; 63.6%) were resistant to [amoxicillin as well as amoxicillin + clavulanic acid], ceftriaxone, and sulfamethoxazole + trimethoprim, respectively. All the Staphylococcus aureus (S. aureus) isolates were resistant to cefoxitin, which implies methicillin-resistant S. aureus (MRSA). The high prevalence of UTIs and antibiotic resistance revealed in the current study needs public health interventions such as educating the population about preventive measures and the importance of early treatment of UTIs. Our findings also highlight the need to provide UTI screening services for PLHIV, and healthcare providers should adopt antibiotic stewardship programs to promote and ensure their appropriate and judicious use.

Presence of carbapenem resistance in hybrid Escherichia coli pathovars from ready-to-eat fresh-cut fruits in Accra, Ghana.

This study reports the presence of carbapenem-resistant Escherichia coli hybrid pathovars and its prevalence in 200 fresh-cut fruits from Accra. Standard culture methods were used to quantify microbial indicators and E. coli on fresh-cut fruits retailed in formal and informal outlets in Accra. The Kirby-Bauer disc diffusion method was used to determine the antibiotic resistance profile of E. coli, while multiplex PCR was employed to identify the virulence and carbapenem-resistance genes. Escherichia coli prevalence in cut fruits was 17%, with pawpaw, watermelon, and mixed fruit having higher prevalence than pineapple. Of the 34 E. coli isolates from fresh-cut fruits, 44% showed broad resistance to beta-lactam antibiotics, while 5.9% showed carbapenem resistance. The study identified virulence genes associated with all E. coli isolates, including stx1, stx2, escV, and ipaH, of which 97% were hybrid pathovars bearing genes for Shiga toxin-producing E. coli/enteropathogenic E. coli/enteroinvasive E. coli. The carbapenemase gene, blaIMP, was associated with both carbapenem-resistant E. coli phenotypes identified. Despite a low-carbapenem-resistance prevalence observed among E. coli isolates, hypervirulent hybrid strains of E. coli is present in fresh-cut fruits in the sampling area, posing a potential public health risk to fresh-cut fruit consumers.

In-Vitro Proliferation and Enrichment of CD34+ Autologous Haematopoietic Stem Cells, Obtained from Patients with End-Stage Left Ventricular Failure

With a rising global prevalence of end-stage heart failure and a limited availability of cardiac transplantation programmes or left ventricular assist devices in many parts of the world, there is a renewed interest in potential alternative treatment options.Along with other groups, we have previously demonstrated the beneficial effect of intracoronary autologous bone marrow derived peripheral haemopoietic stem cell transplantation, to improve myocardial contractility in end-stage heart failure.We now assess the ability of these cells to expand in numbers in-vitro, to achieve enrichment of stem cell counts, using standard cell culture methods and flow- cytometric analysis.We demonstrate that with in-vitro culturing (within 3 passages post-harvest), the CD34+ stem cell fraction increases in standard culture media, across multiple samples. However, optimal culture conditions to achieve near pure stem cell populations with rapid cell proliferation, still needs to be defined.

Revolutionizing chronic endometritis diagnosis: real-time polymerase chain reaction unveils microbial pathogens in Indian women with abnormal bleeding and reproductive challenges

BackgroundThis study aimed to assess the utility of real-time-polymerase chain reaction (PCR) for diagnosing chronic endometritis (CE) by targeting 11 prevalent pathogens and to compare the outcomes with conventional culture-based diagnosis. Study DesignA retrospective analysis was conducted on 500 patients with clinical conditions such as abnormal bleeding, in vitro fertilization failure, recurrent implantation failure, recurrent miscarriage, and recurrent pregnancy loss. The prevalence of 11 key pathogens associated with CE was evaluated in endometrial biopsy samples. ResultsIn our study, PCR identified 318 cases (63.6%) positive for at least one of the 11 investigated pathogens, while culture-based methods detected 115 cases (23%). Predominant pathogens detected by PCR included Enterococcus faecalis (E. faecalis) (19%), Escherichia coli (E. coli) (6.8%), Staphylococcus aureus (S. aureus) (9%), Mycoplasma hominis (5%), Mycoplasma genitalium (6.2%), Streptococcus agalactiae (S. agalactiae) (4.2%), Ureaplasma urealyticum (4%), nontuberculous Mycobacterium (5.2%), Mycobacterium tuberculosis (1.2%), Neisseria gonorrhoeae (0.6%), and Chlamydia trachomatis (2.4%). Standard culture methods identified E. faecalis (10.8%), S. aureus (6.2%), E. coli (3.8%), and S. agalactiae (2.2%). ConclusionThe DICE panel proves itself as a swift, precise, and cost-effective diagnostic tool for detecting both culturable and nonculturable endometrial pathogens in CE. Demonstrating superiority, the Molecular method outshines microbial culture, ensuring accurate and sensitive detection of CE-associated pathogens, harmonizing seamlessly with histology and hysteroscopy findings.

Recombinase polymerase amplification combined with lateral flow biosensor for rapid visual detection of Clostridium perfringens in chicken meat and milk.

Clostridium perfringens is one of the major anaerobic pathogen causing food poisoning and animal enteritis. With the rise of antibiotic resistance and the restrictions of the use of antibiotic growth promoting agents (AGPs) in farming, Clostridium enteritis and food contamination have become more common. It is time-consuming and labor-intensive to confirm the detection by standard culture methods, and it is necessary to develop on-site rapid detection tools. In this study, a combination of recombinase polymerase amplification (RPA) and lateral flow biosensor (LFB) was used to visually detect C. perfringens in chicken meat and milk. Two sets of primers were designed for the plc gene of C. perfringens, and the amplification efficiency and specificity of the primers. Selection of primers produces an amplified fragment on which the probe is designed. The probe was combined with the lateral flow biosensor (LFB). The reaction time and temperature of RPA-LFB assay were optimized, and the sensitivity of the assay was assessed. Several common foodborne pathogens were selected to test the specificity of the established method. Chicken and milk samples were artificially inoculated with different concentrations (1 × 102 CFU/mL to 1 × 106 CFU/mL) of C. perfringens, and the detection efficiency of RPA-LFB method and PCR method was compared. RPA-LFB can be completed in 20 min and the results can be read visually by the LFB test strips. The RPA-LFB has acceptable specificity and the lowest detection limit of 100 pg./μL for nucleic acid samples. It was able to stably detect C. perfringens contamination in chicken and milk at the lowest concentration of 1 × 104 CFU/mL and 1 × 103 CFU/mL, respectively. In conclusion, RPA-LFB is specific and sensitive. It is a rapid, simple and easy-to-visualize method for the detection of C. perfringens in food and is suitable for use in field testing work.

Granulicatella adiacens: An uncommon pathogen in prosthetic joint infections

Granulicatella adiacens is an uncommon culprit in prosthetic joint infections, especially when predisposing factors are absent. Here, we present the case of a 78-year-old female who developed an acute surgical site infection shortly after undergoing left total knee arthroplasty. Despite initial negative cultures, subsequent analysis identified Granulicatella adiacens in the knee fluid, marking only the second reported instance in the United States and the seventh globally. This singular case highlights the considerable challenges associated with isolating this organism, emphasizing its atypical presentation and diagnostic elusiveness. Typically associated with endocarditis and bacteremia, its involvement in isolated surgical site infections is particularly unusual. Our case underscores the necessity for heightened clinical suspicion and the adoption of specialized diagnostic methodologies to accurately detect Granulicatella adiacens infections. Given its fastidious nature and nutritional requirements, standard culture methods may fail to identify this pathogen, necessitating alternative diagnostic approaches. As such, the utilization of specialized media becomes paramount in clinical diagnostics to facilitate appropriate management and optimize healthcare resource allocation. By enhancing our ability to detect and treat infections caused by Granulicatella adiacens, we can mitigate patient morbidity and reduce the burden of unnecessary interventions, thereby improving patient outcomes and healthcare efficiency.

Occurrence Autosomal Mosaicism in Karan Fries Cattle and Murrah Buffalo with Poor Reproduction

Background: The present study was conducted at the Livestock Genome Analysis Laboratory (LGAL), ICAR-National Dairy Research Institute (NDRI), Karnal, Haryana, India, to explore, chromosomal anomalies in reproductively inefficient animals. Chromosomal abnormalities are deviations in normal genetic architecture and lead to disorders in bearer individuals. These can be both in number and structure of autosomes or sex chromosomes, usually inflict adverse effects on anatomy, survival and reproductive performance in domestic animals. Methods: Present report is on a female Murrah buffalo (Bubalus bubalis) and a Karan Fries cattle which were identified during a routine cytogenetic reproductively inefficient females at cattle yard, National Dairy Research Institute, Karnal. Metaphase chromosome preparations were prepared using standard lymphocyte culture method. Slides were stained with Giemsa and treated for R-banding using fluorochrome-photolysis-Giemsa technique (RB-FPG technique). Karyotypes were prepared and abnormal chromosome was identified. Result: Routine examination using Giemsa-stained slides revealed two cell lines with 60 and 61 chromosomal constitutions in cattle and 3 cell lines in buffalo. Among 182 metaphase plates examined were examined for Karan fries which revealed two types of chromosome configuration, the frequency of cells carrying 60 and 61 chromosomes was 83.5% and 16.5%, respectively. R banding technique distinguished autosome 16 responsible for mosaicism. Cytogenetic evaluation of 407 metaphase spreads of Murrah buffalo revealed three types of chromosome configurations, viz. 49, 50 and 51 all with XX and the overall frequency was 16.7, 76.7 and 6.6 per cent, respectively. The R- bandings confirmed autosome 11 both in monosomy (2n=49) and trisomy (2n=51), respectively. This unusual chromosomal number might have arisen due to non-disjunction during early stage of zygotic development of the buffalo and cattle.

Bacillus and Aspergillus Prevalence: an Investigation into the Indoor Microbial Quality of Built Environment at the University of Port Harcourt: Nigeria

Background: Inadequate indoor air quality poses significant challenges in school environments, stemming from factors such as overcrowded classrooms, insufficient ventilation, and substandard construction and maintenance practices. Bacterial and fungal contaminants in indoor air present serious health risks. Methods: This institutional-based cross-sectional study focused on selected learning halls to assess bacterial and fungal loads using the settle plate method. Analysis was accomplished through standard cultural methods. Nutrient agar and Potato Dextrose agar plates were exposed for 15 minutes to capture bacteria and fungi, respectively. Results: The study revealed a total bacterial load of 8300 CFU/m3 in the morning and 10400 CFU/m3 in the evening. The highest bacterial load occurred at MBS 25 (3200 CFU/m3 in the morning and 3700 CFU/m3 in the evening), while the lowest was recorded at the Arena (310 CFU/m3 in the morning and 990 CFU/m3 in the evening). Similarly, fungal loads varied, with the Arena recording the highest (620 CFU/m3 in the morning and 690 CFU/m3 in the evening) and the Faculty of Science Auditorium the lowest (310 CFU/m3 in the morning and 620 CFU/m3 in the evening). Bacillus spp. dominated among bacterial genera (25.9%), while Aspergillus spp. was the most prevalent fungal genus (41.7%). Other bacterial genera included Klebsiella, Staphylococcus, Enterococcus, E. coli, and Pseudomonas, while Penicillium and Candida were among the fungal genera. Conclusion: Mitigation strategies should target factors such as temperature, occupancy, and humidity to curb bacterial and fungal proliferation in indoor lecture halls, thus safeguarding the health of students and teachers in the University of Port Harcourt and similar educational institutions.

Raw meat-based diet for pets: a neglected source of human exposure to Salmonella and pathogenic Escherichia coli clones carrying mcr, Portugal, September 2019 to January 2020.

BackgroundThe pet industry is expanding worldwide, particularly raw meat-based diets (RMBDs). There are concerns regarding the safety of RMBDs, especially their potential to spread clinically relevant antibiotic-resistant bacteria or zoonotic pathogens.AimWe aimed to investigate whether dog food, including RMBD, commercially available in Portugal can be a source of Salmonella and/or other Enterobacteriaceae strains resistant to last-line antibiotics such as colistin.MethodsFifty-five samples from 25 brands (21 international ones) of various dog food types from 12 suppliers were screened by standard cultural methods between September 2019 and January 2020. Isolates were characterised by phenotypic and genotypic methods, including whole genome sequencing and comparative genomics.ResultsOnly RMBD batches were contaminated, with 10 of 14 containing polyclonal multidrug-resistant (MDR) Escherichia coli and one MDR Salmonella. One turkey-based sample contained MDR Salmonella serotype 1,4,[5],12:i:- ST34/cgST142761 with similarity to human clinical isolates occurring worldwide. This Salmonella exhibited typical antibiotic resistance (bla TEM + strA-strB + sul2 + tet(B)) and metal tolerance profiles (pco + sil + ars) associated with the European epidemic clone. Two samples (turkey/veal) carried globally dispersed MDR E. coli (ST3997-complexST10/cgST95899 and ST297/cgST138377) with colistin resistance (minimum inhibitory concentration: 4 mg/L) and mcr-1 gene on IncX4 plasmids, which were identical to other IncX4 circulating worldwide.ConclusionSome RMBDs from European brands available in Portugal can be a vehicle for clinically relevant MDR Salmonella and pathogenic E. coli clones carrying genes encoding resistance to the last-line antibiotic colistin. Proactive actions within the One Health context, spanning regulatory, pet-food industry and consumer levels, are needed to mitigate these public health risks.

Prevalence and antimicrobial susceptibility of Salmonella enteritidis and Salmonella typhimurium isolated from hen eggs and quail eggs in Karaj, Iran.

Different Salmonella serotypes are considered one of the most important food pathogens in the world. Poultry meat and eggs are the primary carriers of Salmonella in human populations. This study aimed to estimate the Salmonella enteritidis and Salmonella typhimurium contamination rates of retail hen and quail eggs in Karaj, Iran. Moreover, the antimicrobial resistance patterns of the strains were evaluated, and the efficiency of the standard culture method and multiplex polymerase chain reaction (m-PCR) were compared. In this descriptive cross-sectional study over 1 year (Jan-Dec 2022), 150 commercial and 150 backyard hen eggs and 300 commercial quail eggs, without cracks and fractures, were collected randomly from best selling groceries in Karaj city. All samples were examined for Salmonella contamination independently by standard culture and m-PCR approaches. A standard disc diffusion method was employed to assess the antimicrobial susceptibility of the strains against 18 antimicrobial agents. Out of 300 examined eggs, 2 S. enteritidis strains were isolated from the shell of backyard hen eggs. The same serotype was also detected in the contents of one of these two eggs. One S. typhimurium was isolated from the shell of a commercial hen egg. Overall, the Salmonella contamination of the shell and contents was 1% and 0.3%, respectively. Salmonella was not isolated from the eggshells or the contents of the quail eggs. There was complete agreement between the results of m-PCR and the standard culture methods. Among the 18 tested antibiotics, the highest resistance was recorded for colistin (100%), followed by nalidixic acid (75%). As most Salmonella spp. are associated with human food poisoning, continuous surveillance is required to effectively reduce the risk posed by contaminated poultry eggs. Furthermore, mandatory monitoring of antimicrobial use on Iranian poultry farms is recommended.

Association between blood procollagen III N-terminal propeptide, granulocyte-macrophage colony-stimulating factor and triple therapy in single inhaler efficacy for chronic obstructive pulmonary disease re-exacerbation prevention

Triple therapy with inhaled corticosteroid (ISC) / long-acting β2 agonist (LABA) / long-acting muscarinic antagonist (LAMA) in single inhaler expanded the possibilities for prevention of chronic obstructive pulmonary disease (COPD) exacerbations. Heterogeneity of COPD determines the needs in search of target population and efficacy markers for each existing therapy. Disease phenotype depends on a complex of factors, with respiratory viral infection among the most significant. Aim of the study was to assess the efficacy of triple therapy with ICS/LABA/LAMA in single inhaler for subsequent COPD exacerbations prevention and to search molecular markers of the efficacy depending the etiology of index exacerbation. Material and methods. It was a prospective observational study of three COPD patients’ strata: after COPD exacerbation required hospitalization with viral (n = 60), bacterial (n = 60) and viral-bacterial (n = 60) infection. Triple therapy in single inhaler (n = 104) or in free combinations (n = 76) were prescribed in real clinical practice. COPD was diagnosed according to spirography criteria. To establish the COPD exacerbation etiology the real time PCR of sputum or bronchoalveolar lavage fluid, standard cultural method, blood procalcitonin, as well as marker blood proteins, hyaluronic acid by ELISA measurement were done. Associations were revealed using Cox regression. Results. Triple therapy in single inhaler in comparison with free combinations decreased time to first re-exacerbation, hazard ratio (HR) in viral-associated index exacerbation strata was 0.38 (95% confidence interval (95% CI) 1.15–0.40), in bacterial – 0.47 (0.39–0.72), in viral-bacterial – 0.39 (0.14–0.39). In strata of COPD patients after viral and viral-bacterial exacerbations, in subgroups treated with triple therapy in single inhaler blood procollagen III N-terminal propeptide (PIIINP) (HR for group after viral index exacerbations was 1.03, 95 % CI 1.02–1.28, HR for group after viral-bacterial exacerbations was 1.04, 95 % CI 1.02–1.28), granulocyte-macrophage colony-stimulating factor (GM-CSF) (HR 1.03, 95 % CI 1.02–1.32, 1.01, 95 % CI 1.00–1.35, respectively) content was associated with time of re-exacerbations. Conclusions. Blood PIIINP and GM-CSF during COPD exacerbation are perspective markers of subsequent exacerbations within 1 year in patients after virus-associated or viral-bacterial index exacerbation. In these groups of patients triple therapy in single inhaler is more effective than free combination for subsequent exacerbations prevention.

Comparative performance of biofire pneumonia panel and standard culture-based methods for diagnosing pneumonia in critically ill patients: Impact on antibiotic stewardship

IntroductionLower respiratory tract infections (LRTIs) are a common cause of morbidity and mortality worldwide. Accurate identification of the pathogens causing LRTIs is crucial for ensuring of diagnostic and antibiotic stewardship. The Biofire Pneumonia Panel (BFPP) is a molecular diagnostic test that allows rapid detection of various bacterial and viral pathogens. In this study, we compared the performance of BFPP with standard culture methods for the detection of pathogens. Materials and methodsRespiratory samples from 70 patient with suspected LRTIs were tested using both BFPP and standard culture methods. The distribution of isolated bacterial pathogens was analyzed, and the sensitivity and specificity of BFPP were calculated. Additionally, the performance of BFPP in detecting antimicrobial resistance genes was evaluated. The results were compared with those obtained from VITEK-2 antimicrobial susceptibility testing and culture-based methods. ResultsAmong the suspected LRTI cases, BFPP identified a single pathogen in 32.8% of cases and multiple pathogens in 40% of cases. The standard culture method detected a single pathogen in 47.1% of cases. BFPP showed a sensitivity of 93.9% and a specificity of 45.9% for the total sample. The performance of BFPP in detecting antimicrobial resistance genes varied for different pathogens with overall sensitivity of 40.1% and specificity of 95.9%. ConclusionThe Biofire Pneumonia Panel (BFPP) demonstrated high sensitivity for several bacterial pathogens, indicating its potential as a rapid diagnostic tool. However, its performance varied for different microorganisms, and it had limitations in detecting certain pathogens and antimicrobial resistance genes for which still required more further studies to explore different resistance gene mechanism that can be incorporated in this panel in future. The BFPP can complement standard culture methods as a rapid tool in the diagnosis of LRTIs.

Molecular Characterization of Plasmid-mediated Quinolone Resistant Salmonella typhi From Patients Attending Federal Medical Center, Jabi, Abuja, Nigeria

Aims: This study investigates and reports the detection of qnr genes (plasmid-mediated quinolone resistance PMQR) in S.typhi isolated from stool of patients with suspected typhoid fever, in Federal Medical Center, Jabi, Abuja, Nigeria. Study Design: Cross sectional study. Place and Duration of Study: Department of Microbiology, Nasarawa State University, Keffi, between October 2022 and November 2023. Methodology: Salmonella typhi was isolated from stool of patients with suspected typhoid fever using standard culture and microbiological methods. Antibiotic susceptibility testing were performed using the disc diffusion method to investigate the ability of S. typhi to resist some antibiotics. Qnr genes (qnrC,qnrD,qnrS) were detected by PCR and amplification. Results: 17 out of 150 (11.3%) of the samples collected had S.typhi. Antibiotic resistance in the isolates in decreasing order were as follows: cefotaxime (100.0%), amoxicillin/ clavulanic acid (94.1%), nalidixic acid (94.1%), cefuroxime (94.1%), imipenems (88.2%), ceftriaxone/sulbactam (82.4%), ciprofloxacin (82.4%), gentamicin (58.8%), levofloxacin (47.1%) and ofloxacin (29.4%).The commonest antibiotic resistant phenotype was AUG- CTX-IMP-OFX-CN-NA-CXM-CRO-CIP-LBC at 29.4%. Multiple antibiotic resistance (MAR) was observed in 100% (17/17) of the isolates with the common MAR indices being 1.0 (29.4%), 0.7 (23.5%), 0.8 (17.6%) and 0.5 (17.6%). 33.3% of the isolates in FMCJ were positive for qnrC,qnrD,qnrS,qnrA+qnrC and qnrS+ aac(6′)-Ib-cr as all of qnrC qnrD,and qnrS genes were all expressed in the isolates. Conclusion: The S. typhi isolates showed lower resistances to ofloxacin, levofloxacin, and gentamicin, and all isolates were MAR, with resistance to 10 antibiotics being the most predominant. In addition, qnrC,qnrD, and qnrS resistance genes were all expressed in the isolates.