Frontiers in Cellular and Infection Microbiology

IntroductionUrinary tract infections (UTIs) are among the most encountered bacterial infection of humans and affect both male and female of all age groups, resulting in high mortality of infected patients if not properly managed. Hypothesis/Gap statementSeveral studies in different sub-Saharan Africa locations show variations in incidence of UTI as well as its causative organisms. AimThis study aimed to assess the prevalence, etiological agents, and factors associated with urinary tract infections among patients attending selected hospitals in Calabar metropolis, Nigeria. MethodologyThis was a cross-sectional study. Mid-stream urine samples collected from 240 patients with UTI were cultured using Cystine Lactose Electrolyte Deficient Agar (CLED). Data on socio-demographic, clinical symptoms and risk factors were obtained using structured questionnaire. Uropathogens were characterized using microbiological and biochemical tests and confirmed with API(R) 20E and 20NE (Biomerieux) identification system. Pearson Chi-square was employed to check associations between categorical variables. P-value of <.05 was considered statistically significant. ResultsOut of the 240 urine samples collected, 13 were contaminated during collection, and 227 analyzed. Sixty-five (28.6%) patients had significant bacteriuria. Previous history of UTI (P=.000, CI=1.582-5.180), use of drugs without prescription (P=.000, CI=0.040-0.220), pregnancy (P=.001, CI=1.858-12.575) and history of urinary catheter (P=.031, CI=1.024-19.053) showed significant association with the occurrence of UTI. Klebsiella pneumoniae (23.1%) was the most predominant isolate, followed by Coagulase-negative Staphylococci (16.9%) and Escherichia coli (12.3%). ConclusionThe study shows that routine UTI screening is beneficial for pregnant women, patients with difficult or painful urination, patients with previous episodes of UTI, catheterized patients and appropriate drugs administered for positive cases. In addition, self-administration of antibiotics should be avoided.

Lyme disease (LD) is an illness caused by the spirochete Borrelia burgdorferi (B. burgdorferi). Borrelia is known to disseminate through organs, including the skin, joints, spinal cord, bladder, and heart, leading to Lyme arthritis, neuroborreliosis, and Lyme carditis. While previous studies have investigated the impact of LD on pregnancy in both mice and humans and have found the presence of B. burgdorferi in the uterus of mice, we studied the impact of LD on the non-pregnant female reproductive tract. We use a mouse model for LD and find an ongoing and severe infection of the reproductive tract of female mice, which persists up to 15-months post-inoculation. This infection results in uterine glandular cysts and endometrial hyperplasia as well as vaginal epithelial thickening, polymorphonuclear and mononuclear cell epithelial infiltration, and epithelial desquamation into the vaginal lumen. Strikingly, we find that age has an impact on the extent of gynecologic pathology such that aged female mice (1-year old) that are reproductively senescent have more gynecologic pathology with infection compared to young mice (15-weeks old) when infected for the same length of time. Using large-scale electronic healthcare record data, we report that LD additionally results in increased infection-associated risk of menorrhagia (1.5-fold), miscarriage (1.62-fold), uterine fibroids (1.42-fold), and endometriosis (1.93-fold). Underreporting of gynecological outcomes is pervasive throughout many different infectious diseases, and LD-associated gynecological pathologies may have been similarly underappreciated in the field. This work suggests that further study of the female reproductive tract and the effects of B. burgdorferi infection therein will help clarify and expand the knowledge of myriad LD outcomes. Graphical abstract O_FIG O_LINKSMALLFIG WIDTH=149 HEIGHT=200 SRC="FIGDIR/small/25323258v1_ufig1.gif" ALT="Figure 1"> View larger version (59K): org.highwire.dtl.DTLVardef@1eac89corg.highwire.dtl.DTLVardef@1189778org.highwire.dtl.DTLVardef@1807b22org.highwire.dtl.DTLVardef@140e91_HPS_FORMAT_FIGEXP M_FIG C_FIG

Urinary tract infections (UTI) are one of the most common infections, worldwide. To understand mechanisms of UTI pathogenesis and find new treatments, researchers often use animal models, such as mice or rats. However, studying certain phenotypes in animals can be difficult. Additionally, using animals in research comes with significant administrative and ethical challenges. To address these challenges, we developed a simple, reproducible, and cost-effective model to study UTI using donated mouse bladder tissue that would otherwise be discarded. This model allows researchers of all experience levels to study interactions between the host and pathogen in a controlled environment. We tested uropathogenic E. coli colonization and invasion of isolated urothelial sheets from 30 minutes to 24 hours, finding that bacterial burden in our ex vivo model was comparable to in vivo UTI mouse models. To optimize reproducibility, we tested multiple variables, including technical parameters, such as incubator conditions, and biological factors, such as biological sex or prior pregnancy in the donor mouse. This method offers several advantages, including assessment of early host-pathogen interactions, immune cell uptake of bacteria, the impact of age and sex of donor animals in infection, and diverse bacterial strains, mutants, or treatments. In addition, in some countries, sharing material recovered from animals sacrificed for other reasons does not require additional ethical approval by the receiving laboratory, providing a resource for labs without access to animals and reducing administrative burden. Given the breadth of the model with respect to sex, age, mouse and bacterial strain, and the ability to test any parameter that can be included in a 96-well plate, we believe this model will be useful to UTI researchers, with potential application beyond infection or even beyond the bladder to other tissues.

Urinary tract infections (UTIs) are the most common infections in both outpatient and inpatient settings, contributing significantly to morbidity, reduced quality of life, and antimicrobial overuse. Although urine culture remains the diagnostic gold standard, it poses practical limitations in clinical workflows. Rapid diagnostic methods such as urine flow cytometry (UF) offer potential for timely, reliable UTI detection. We conducted a multicenter diagnostic study to evaluate the performance of BACT-Info. and UTI-Info. flags on the UF-5000/4000 system in detecting UTIs, using presumptive Gram staining and urine culture (>10 CFU/mL) as references. A total of 763 patients with suspected UTI were enrolled, and 721 patients--with uropathogenic bacteria and complete data--were included in the final analysis (384 with culture-confirmed UTI and 337 without UTI). The diagnostic value of nitrituria was highly specific, while leukocyte esterase was sensitive. For BACT-count and WBC-count of UF-5000/4000, the AUCs were 0.85 and 0.69, respectively. Using cutoffs of WBC >82.05/{micro}L or bacteria >975.4/{micro}L, the UTI-Info flag demonstrated 89% sensitivity, 54% specificity, 69% positive predictive value, and 82% negative predictive value. Positive and negative likelihood ratios were 1.93 and 0.20, respectively. The agreement between the BACT-Info. flag and Gram typing showed Kappa values of 0.716 for Gram-negative and 0.216 for Gram-positive bacteria when compared to culture, and 0.721 and 0.401, respectively, when compared to presumptive Gram staining. The UF-5000/4000 UTI-Info. and BACT-Info. flags show promise as a rapid UTI screening tool. The combination of these flags with urinalysis parameters, nitrite testing and leukocyte has potential to be developed into a diagnostic algorithm for early and sensitive UTI prediction. Such an approach may reduce unnecessary urine cultures and support timely, appropriate empiric antibiotic therapy. Establishing optimal cutoffs tailored to specific clinical settings is essential to enhance diagnostic accuracy and improve clinical utility.

BackgroundVulvovaginal Candidiasis is a condition commonly caused by Candida albicans. It is the second most common infection of the female genitalia affecting many women worldwide. Studies have identified unhealthy genital care practices associated with the infection among women including expectant mothers. Knowledge of the various signs and symptoms is crucial for early detection, reporting, and treatment. Good knowledge may influence healthy practices limiting the infection and its complications. This study assessed the relationship between knowledge, practices and occurrence of Vulvovaginal candidiasis among pregnant women accessing antenatal care at a teaching hospital in Ghana. MethodsA cross-sectional study was conducted among 336 pregnant women receiving antenatal care at the Ho Teaching Hospital. A structured questionnaire was employed in assessing their knowledge on the infection and some practices regarding vaginal hygiene. Hospital records of these participants were further checked to verify the occurrence of the infection among them. Analysis to identify associations between outcome variables and risk factors as well as significance level was carried out. ResultsOut of the 336 gestational mothers involved in the study, 27% were found to have been diagnosed with candidiasis at the time of the study. Pregnant women who usually use antibiotics had 2.25 increased odds of developing Vulvovaginal Candidiasis (VVC) compared to those who do not [OR:2.25 95CI:1.33-3.79; p-value = 0.003]. Again, a greater percentage of the study participants, 85% had good knowledge whiles 5% had poor knowledge. ConclusionThe occurrence of VVC was elevated in the study jurisdiction. Frequent antibiotic use was found as a significant factor associated with the occurrence of the infection.

BackgroundUrinary tract infections caused by Extended Spectrum Beta-Lactamase producing Escherichia coli are increasing globally and yet treatment still remains a challenge due to antibiotic resistance of the causative agent. ObjectivesThe aim of the study was to determine the antimicrobial susceptibility pattern and detect the presence of blaCTX-M gene in Escherichia coli isolated from urinary tract infection patients at the University Teaching Hospital, Lusaka, Zambia. MethodologyThis was a cross sectional study that involved collection of urine samples from patients who were diagnosed with urinary tract infections. The samples were cultured on MacConkey agar complemented with cefotaxime and Polymerase Chain Reaction was performed to confirm the Extended Spectrum Beta-Lactamase producers by detecting the CTX-M gene. Antimicrobial susceptibility tests were conducted using standard methods. ResultsA total of 327 urine samples were cultured and 15 (4.6%) of these samples were positive ESBL producers. The isolates showed complete resistance to ampicillin and cotrimoxazole. ConclusionMulti drug resistant Extended Spectrum Beta-Lactamase producing Escherichia coli was detected in 4.6 % of UTI patients at the University Teaching Hospital.

Urinary tract infections (UTIs) affect 405 million people worldwide. Current diagnostics for UTIs rely on cultures, which can take 2 to 4 days. This study evaluated eleven culture independent methods for sample preparation from 77 complicated UTI patient samples, followed by real-time nanopore sequencing and data analysis. The metagenomic results were highly consistent with culture-based clinical routines. The optimized in-house method demonstrated a mean accuracy score of 99% for pathogen identification and 90% for antimicrobial susceptibility profiling. The methods robustness is highlighted by accurately identifying the pathogen with as few as 33 bacterial cells/{micro}L and a low bacterial-to-leukocyte ratio limit of 0.04. Additionally, mNGS identified 13 pathogens that routine diagnostics missed, which were subsequently confirmed by Vivalytic or PCR. This method is up to 30% cheaper than published studies and commercial kits. AUROC analysis indicated that DNA yield and flow cytometry can be used for pre-screening to reduce costs, which is crucial for clinical adoption. This research highlights the rapid diagnosis of UTIs in clinical settings using a cost-effective and scalable method that requires four hours from sample collection to informed decision making. Furthermore, it aims to improve antimicrobial and diagnostic stewardship by reducing empirical treatment and ensuring more judicious antibiotic use.

The human gastrointestinal tract (GIT) is home to an abundance of diverse microorganisms, and the balance of this microbiome plays a vital role in maintaining a healthy GIT. The obstruction of the flow of bile into the duodenum, resulting in obstructive jaundice (OJ), has a major impact on the health of the affected individual. This study sought to identify changes in the duodenal microbiota in South African patients with OJ compared to those without this disorder. Mucosal biopsies were taken from the duodenum of nineteen jaundiced patients undergoing endoscopic retrograde cholangiopancreatography (ERCP) and nineteen control participants (non-jaundiced patients) undergoing gastroscopy. DNA extracted from the samples was subjected to 16S rRNA amplicon sequencing using the Ion S5 TM sequencing platform. Diversity metrics and statistical correlation analyses with the clinical data were performed to compare duodenal microbial communities in both groups. Differences in the mean distribution of the microbial communities in the jaundiced and non-jaundiced samples were observed; however, this difference did not reach statistical significance. Of note, there was a statistically significant difference between the mean distributions of bacteria comparing jaundiced patients with cholangitis to those without. On further subset analysis, a significant difference was observed between patients with benign (Cholelithiasis) and malignant disease, namely head of pancreas (HOP) mass (p-values of 0.01). Beta diversity analyses further revealed a significant difference between patients with stone and non-stone related disease when factoring in the Campylobacter-Like Organisms (CLO) test status (p=0.048). This study demonstrated a shift in the microbiota in jaundiced patients, especially considering some underlying conditions of the upper GI tract. Future studies should aim to verify these findings in a larger cohort.

BackgroundCatheter-associated urinary tract infections (CAUTIs) are a prevalent healthcare-associated infection, accounting for significant morbidity, mortality, and increased healthcare costs. MethodThis is a cross-sectional study of patients diagnosed with UTI associated with catheter use. The sample was collected from November 2023 to June 2024, consisting of 200 patients admitted to the surgical, medical, and trauma wards of tertiary hospitals in Karachi, namely Jinnah Postgraduate Medical Centre Karachi and Dr. Ruth K. M. Pfau Civil Hospital Karachi. Data is analyzed using SPSS Version 22 and P-value of 0.05 considered significant. ResultThe majority of respondents (59.5%) had their catheters changed since insertion, predominantly by trained nurses (93.0%). There were notable associations with underlying conditions such as hypertension (56.5%) and diabetes (44.5%). Gender differences were significant, with females leading in medical cases and males in surgical and trauma cases (p-value 0.017). Age-related trends showed the 55+ age group dominated medical cases, while surgical and trauma cases varied by age group. There was a significant relationship between bleeding during catheterization and UTI (p-value: 0.000). ConclusionThe study revealed a minimal incidence of CAUTI in Karachis tertiary care hospitals, indicating effective practices. However, further research is needed to explore the potential risk factors identified, such as female gender and comorbidities, to develop targeted interventions for reducing CAUTI incidence and improving patient outcomes.

Antimicrobial resistance (AMR) is one of the most dangerous global threats since antimicrobial discovery. The world health organization(WHO) has implemented a program called GLASS to mitigate resistance across the globe. Urinary tract infection(UTI) are the second most common infections and are the most common reason for prescription of antimicrobials, the rise in AMR has caused concerns of UTI Overuse and misuse of prescriptions and decrease of treatment options hence many researches conducted across the globe are on uropathogens resistance rate and trend. This retrospective study was conducted in duhok province of KRI to measure antimicrobial resistance percentages and identify the most common uropathogens. 309 urine samples were collected in a time span of 12 months. Urine samples were collected by clean catch midstream and inoculated on blood and MacConkey agars, Antibiotic sensitivity test (AST) was performed to identify Gram negative uropathogen and its sensitivity pattern. We found out most common Gram negative uropathogen in females were E.coli and Klebsiella pneumonia while in males it was E.coli and Pseudomonas aeruginosa and common Klebsiella pneumonia. E.coli was most resistance to amoxicillin/amp(64.2%) and it was least resistant to carbapenems(6.1%). Klebsiella pneumonia had similar resistant pattern to E.coli. pseudomonas aeruginosa was highly resistant to all antimicrobials, third gen cephalosporins were the highest 95.7%. AMR has risen to concerning levels in duhok and if not controlled would result in simple infections causing death in future we recommend guidelines for control of Overuse, misuse and ease of availability of antimicrobials as a measure to decrease AMR. Continues monitoring should be performed on AMR development in the future.

Chagas disease is caused by infection with the protozoan parasite Trypanosoma cruzi. Despite triggering a strong immune response, infections are typically life-long and can result in severe cardiac and/or digestive tract pathology. Current drugs have limited efficacy, and treatment failure is a common outcome. Eliminating a non-replicating T. cruzi sub-population that can persist after therapy has been a key challenge for the drug-development community. Here, we describe the transcriptome and proteome profiles of quiescent epimastigote forms of the parasite isolated from exponentially growing cultures on the basis of reduced turnover of transiently-induced red fluorescent protein. This quiescent sub-population was characterised by down-regulation of genes/proteins involved in translation, metabolism, mitochondrial function and DNA replication, and by up-regulation of proteins that promote exit from the cell-cycle in other organisms. These data represent a resource that can be exploited to dissect the mechanistic basis of quiescence and to refine the drug-development screening cascade.

Pathogenic Leptospira species can survive and thrive in a wide range of environments. Distinct environments expose the bacteria to different temperatures, osmolarities, and amounts and sources of nutrition. However, leptospires are mostly cultured, in a laboratory setting under in vitro conditions that do not reflect natural environments. This constraint on laboratory cultures limits the applicability of in vitro studies to the understanding of even simple pathogenic processes. Here we report, investigate, and identify a medium and conditions that mimic the host environment during leptospirosis infection, expanding the available in vitro tools to evaluate leptospiral pathogenesis. We quantified genome-wide gene expression of pathogenic Leptospira interrogans cultured in different in vitro media compositions (EMJH, DMEM, EMEM, and HAN). Using EMJH as standard, we compared gene expression in these compositions to genome-wide gene expression gathered in a host environment: whole blood (WB) of hamsters after infection with pathogenic leptospires. Leptospires cultured in DMEM and EMEM media shared 40% and 47% of all differentially expressed genes (DEGs) of leptospires present within WB (FDR<0.01), while leptospires cultured in HAN media only shared 20% of DEGs with those from WB. Furthermore, gene and pathway expression of leptospires cultured on DMEM and EMEM media exhibited a better correlation with leptospires grown in WB, including promoting expression of a similar leptospiral lipid A profile to the one identified directly in host tissues. Taken together, these results indicate that commercial cell-culture media EMEM or DMEM are better surrogates for in vivo pathogenic studies than EMJH or HAN media in Leptospira. These alternative culture conditions, using media that are a standard supply worldwide, provide a reproducible and cost-effective approach that can accelerate research investigation and reduce the number of animal infections necessary for basic research of leptospirosis.

BackgroundThere is an increase in the prevalence of Helicobacter pylori infection in Sudan, accompanied by a high incidence of upper gastrointestinal malignancy. The cytotoxin-associated gene cagA gene is a marker of a pathogenicity island (PAI) in H. pylori and plays a crucial role in determining the clinical outcome of Helicobacter infections. ObjectiveThis study aimed to determine the frequency and heterogeneity of the cagA gene of H. pylori and correlate the presence of cagA gene with clinical outcomes. Materials and methodsFifty endoscopy biopsies were collected from Fedail and Soba hospitals in Khartoum state. DNA was extracted using the Guanidine chloride method followed by PCR to amplify 16S rRNA and cagA gene of H. pylori using specific primers. DNA amplicons of cagA gene were purified and sequenced. Bioinformatics and statistical analysis were done to characterize and to test the association between cagA gene and gastric complications. ResultsCagA gene was detected in 20/37(54%) of the samples that were found positive for H. pylori. There was no association between endoscopy finding and the presence of the cagA gene (p = 0.225). Specific amino acid variations were found at seven loci related to strains from a patient with duodenitis, gastric ulcer, and gastric atrophy (R448H, T457K, S460L, IT463-464VA, D470E, A482Q, KNV490-491-492TKT) while mutations in cancerous strain were A439P, T457P, and H500Y. ConclusionDisease-specific variations of cagA of H. pylori strains, in the region of amino acid residues 428-510, were evident among Sudanese patients with different gastroduodenal diseases. A novel mutation (K458N) was detected in a patient with duodenitis, which affects the positive electrostatic surface of cagA. Phylogenetic analysis showed a high level of diversity of cagA from Sudanese H. pylori strains.

BackgroundSerendipitously, it was observed that fecal transplantation made for Clostridium difficile may cure chronic urinary tract infections. This led us to evaluate the passage in the urine of probiotics contained in yoghurt, which have been claimed to prevent urinary infections. ResultsA commercial yogurt that contained 3 probiotics (Bifidobacterium animalis, Lactobacillus delbrueckiii and Lactobacillus reuteri) was consumed by 28 healthy subjects. We performed by culturomics, urine analysis before and after feeding these yogurts. Genome sequencing of the bacterial strains absent before yogurt consumption and present after consumption was performed. Testing more than 40,000 colonies by MALDI-TOF, we observed in two men and one woman (11% of subjects included), the urine colonization of the same Lactobacillus reuteri present in yogurts, with the same genome with 50 genes never identified in other lactobacilli. ConclusionThis confirms that, as ingested, Lactobacillus salivarius passes into the milk of lactating women, some bacteria, particularly Lactobacillus can colonize body fluids previously considered sterile after ingestion via the digestive route. Although the consequences of this passage remain unknown, we prove for the first time that there is a digestive passage in the urine after consumption of probiotics, including fermented products sold commercially.

BackgroundWhile monosodium urate (MSU) crystal deposition within coronary plaques links hyperuricemia/gout to cardiovascular risk, the existence and functional consequences of MSU deposits within the myocardial interstitium or cardiomyocytes remain unknown. MethodsForensic autopsy myocardial and renal specimens from gout patients (n=4) and controls (n=4) were analyzed for MSU crystals using compensated polarized microscopy, laser capture microdissection coupled with HPLC-MS, and cryo-electron microscopy (cryo-EM) with electron diffraction. Pharmacological (adenine-diet) and genetic (Uox-/-) hyperuricemia murine models were assessed by crystal detection, echocardiography, and correlative histopathology. ResultsMyocardial MSU crystals were identified in all human gout cases (absent in controls), localized perivascularly, interstitially, and intracellularly within cardiomyocytes. Definitive verification was provided by compositional analysis (HPLC-MS m/z 167) and crystal structure confirmation (cryo-EM diffraction matching MSU). Hyperuricemic mice exhibited myocardial MSU deposition, progressive diastolic dysfunction (r =0.7014 vs. crystal burden), and reduced survival associated with higher crystal load. Importantly, myocardial crystal burden correlated more strongly with cardiac impairment than serum uric acid levels. ConclusionThis study provides conclusive morphological, chemical, and crystallographic evidence of intracardiac MSU deposition in gout. It directly links myocardial crystal accumulation to diastolic dysfunction, proposing "uric acid cardiomyopathy" as a novel disease entity. Collectively, these findings achieve a paradigm shift in the perception of gout, redefining it from a peripheral arthropathy to a systemic disorder capable of causing direct myocardial injury.

M. brumae is a rapid-growing, non-pathogenic Mycobacterium species, originally isolated from environmental and human samples in Barcelona, Spain. M. brumae is not pathogenic and its in vitro phenotype and immunogenic properties have been well characterized. However, the knowledge of its underlying genetic composition is still incomplete. In this study, we first describe the 4 Mb genome of the M. brumae type strain ATCC 51384T assembling PacBio reads, and second, we assess the low intraspecies variability by comparing the type strain with Illumina reads from three additional strains. M. brumae genome is composed of a circular chromosome with a high GC content of 69.2 % and containing 3,791 CDSs, 97 pseudogenes, one prophage and no CRISPR loci. M. brumae has shown no pathogenic potential in in vivo experiments, and our genomic analysis confirms its phylogenetic position with other non-pathogenic and rapid growing mycobacteria. Accordingly, we determined the absence of virulence related genes, such as ESX-1 locus and most PE/PPE genes, among others. Although immunogenic potential of M. brumae was proved to be as high as Mycobacterium bovis BCG, the only mycobacteria licensed to treat cancer, the genomic content of M. tuberculosis T cell and B cell antigens in M. brumae is considerably lower than those of M. bovis BCG. Overall, this work provides relevant genomic data on one of the species of the mycobacterial genus with high therapeutic potential.

ObjectiveIn recent years, the incidence of gallstones and their complications has increased, bringing a heavy burden to patients, emphasizing the need to explore the pathogenesis of gallstones. Evidences suggest that the formation of gallstones is closely related to the biliary tract and the gut flora. This study aims to reveal the diversity and abundance of intestinal flora in patients with biliary stones, investigate the relationship between the structure of gallstone formation and its flora, and preliminarily research gene function annotation and metabolic pathways. MethodsThe subjects were 21 eligible gallstone patients undergoing surgery and 20 eligible gallstone-free patients admitted to Beijing Tiantan Hospital, Capital Medical University, from November 2019 to November 2020. Gallstones (GSS group), bile (GSZ group), gallbladder mucosa (GSN group), feces (GSF group) samples were collected from the gallstone group, as well as feces from the control group (HF group). High-throughput sequencing of the V3-V4 regions of the 16S rRNA gene was performed by the Illumina HiSeq platform, bioinformatics analysis was performed on the sequencing results. Results1. The age, body mass index (BMI) and indirect bilirubin (IBil) of gallstone patients were higher than gallstone-free patients (P < 0.05). 2. A total of 23 427 Operational Taxonomic Units (OTUs) were identified in this study, with a mean {+/-} standard deviation of 340{+/-}93, including 4 095 from gallstones (GSS group), 3 065 from bile (GSZ group), 4 687 from gallbladder mucosa (GSN group), and 5 203 from feces (GSF group). 6 377 OTUs were identified from the feces of the gallstone-free control group (HF group). 3. There was no significant difference in the diversity and phylum composition of intestinal flora between gallstone patients and the control group (P > 0.05); however, at the genus level, Achromobacter (P=0.010), Faecalibacterium (P=0.042), Lachnospira (P=0.011) were significantly reduced, while Enterococcus (P=0.001) was significantly increased. 4. The diversity and composition of biliary flora (stone, bile, mucosa) among patients with gallstones have no statistical differences (P > 0.05). The diversity and composition between the biliary and intestinal microflora in gallstones patients have statistical differences: (1) The diversity of biliary flora was significantly higher than the intestinal flora (Simpson index, P < 0.05). (2) At the phylum level, the abundance of Proteobacteria in the bile duct (stone, bile and mucosa) was significantly higher, while Firmicutes and Bacteroidetes were significantly lower than in the intestinal tract (P < 0.05). (3) At the genus level, the abundance of Acinetobacter in the biliary tract was significantly higher, while Bacteroides, Faecalibacterium, Lachnoclostridium and Subdoligranulumbacteria were significantly lower than in the intestinal tract (P < 0.05). 5. The patients stone, bile and gallbladder mucosa shared more than 90% of OTUs. The shared OTUs of intestinal flora between gallstones patients and the control group was greater than 85%, while the five groups of samples shared more than 60% of OTUs. 6. LefSe showed that LDA > 4 in the biliary tract was Gammaproteobacteria, Pseudomonadales, Moraxellaceae, Acinetobacter, Betaproteobacteria, Burkholderiales and Prevotella that all belong to Proteobacteria. ConclusionThe intestinal flora of patients with gallstones and without gallstones exhibited significant bacterial heterogeneity at the genus level. Compared with the intestinal flora of patients with gallstones, the biliary flora exhibited higher diversity. There were significant differences in the bacterial community structure at the phylum and genus levels. The biliary tract (stone, bile, mucosa) and intestinal flora of patients with gallstones have overlaps and differences, which provides the foothold for future studies on the biliary tract flora.

Helicobacter cinaedi infects the human gut and causes invasive infections such as bacteremia and cellulitis through bacterial translocation. However, the mechanism by which H. cinaedi attaches to host cells and establishes infection remains unclear. This study aimed to investigate the relationship between a novel putative autotransporter protein, H. cinaedi autotransporter protein A (HcaA), and its role in pathogenicity. The cytotoxicity of H. cinaedi infection in colon epithelial cell lines (Caco-2 and HT-29) was assessed using a lactate dehydrogenase assay, and it was found that cytotoxicity significantly decreased upon HcaA knockout. Adhesion assays further revealed that the HcaA-knockout strain showed significantly reduced attachment to the human epithelial colorectal adenocarcinoma cell line (Caco-2) compared to that of the wild-type strain. Moreover, the recombinant HcaA protein demonstrated strong adhesion properties to the human monocytic cell line (U937). The adhesive activity was diminished when the RGD motif in HcaA was replaced with RAD, indicating that the RGD motif in HcaA is crucial for host cell adhesion. To determine the role of HcaA in H. cinaedi infection in vivo, C57BL/6 mice were orally infected with wild-type and HcaA-knockout H. cinaedi strains. Bacterial colonization was assessed 7, 14, and 28 days post-infection. At 7 days post-infection, colonization was significantly lower in mice infected with the HcaA-knockout strain compared to those infected with the wild-type strain. In conclusion, our findings suggest that HcaA, a novel putative autotransporter protein in H. cinaedi, plays a significant role as an adhesin in establishing colonization. IMPORTANCEHelicobacter species are classified as gastric or enterohepatic according to their habitat. Among enterohepatic Helicobacter species, which inhabit the intestine, colon and liver, H. cinaedi has been most frequently isolated from humans. H. cinaedi often causes bacteremia and cellulitis in immunocompromised hosts. Here, we focused on the H. cinaedi autotransporter protein A (HcaA), a novel virulence factor in H. cinaedi. We discovered that HcaA contributes to cell adhesion via its RGD motif. Furthermore, in animal experiments, bacterial colonization was reduced in mice infected with HcaA-knockout strains, supporting the hypothesis that HcaA contributes to H. cinaedi adhesion to host cells. Our study provides a novel mechanism for the establishment of H. cinaedi infections and provides new insights into the role of autotransporter proteins in the establishment of Helicobacter infection.

BackgroundHeart failure is a complex clinical syndrome resulting from structural and functional impairment of ventricular filling or ejection of blood. Cor pulmonale is one type of this clinical syndrome. There are only a few published studies on cor pulmonale from Ethiopia. The objective of this study was to determine the prevalence among patients with heart failure and the clinical and laboratory profiles of patients with cor pulmonale who had follow up at one of the tertiary hospitals in Ethiopia MethodsA hospital-based cross-sectional study was conducted at Felege Hiwot Referral Hospital from December 2018 to April 2019. A single population proportion formula was used to determine the number of heart failure patients that had to be included in the study to determine the prevalence of cor pulmonale. The medical records of cor pulmonale patients among the sample heart failure patients were then retrieved and data was extracted using a structured checklist. Data was entered into, cleaned, and analyzed using IBM.SPSS version 23.0. Descriptive statistics were used to report the findings. ResultsEight percent (35) of patients with heart failure had cor pulmonale. Fifty-four point three percent (19) of the patients with cor pulmonale were males and 45.7 %(16) were females. The median age of patients with cor pulmonale was 55 years. The commonest clinical features were cough and dyspnea (91.4 % and 97.1 % respectively). All patients had right ventricular dilation on echocardiography. Pulmonary Complications post-treatment for tuberculosis were the leading causes followed by interstitial lung disease. There was no identified respiratory disease in 40% of patients with cor pulmonale ConclusionCor pulmonale accounted for less than 10 % of heart failure cases. Complications post-pulmonary tuberculosis were found to be the leading respiratory conditions underlying the cor pulmonale. Programs on prevention, early detection, and treatment of pulmonary tuberculosis must be strengthened.

Mycobacterium abscessus (Mab) colonies adopt smooth (S) or rough (R) morphotypes, which are linked to the presence or absence of glycopeptidolipids (GPL), respectively. Though clinically relevant, the association between GPL levels, morphotype and pathogenesis are poorly understood. To investigate the degree of correlation between Mab morphology, GPL levels, and infectivity, we generated isolates from Mab-positive sputum samples from cystic fibrosis patients. Isolated strains were categorised based on their morphology, GPL profile, and replication rate in macrophages. Our findings revealed that around 50% of isolates displayed mixed morphologies and GPL analysis confirmed a consistent relationship between GPL content and morphotype was only found in smooth isolates. Across morphotype groups, no differences were observed in vitro, yet using a high-content THP-1 cell ex vivo infection model, clinical R strains were observed to replicate at higher levels. Moreover, the proportion of infected macrophages was notably higher among clinical R strains compared to their S counterparts at 72 hours post-infection. Clinical variants also infected at significantly higher rates compared to laboratory strains, highlighting the limited translatability of lab strain infection data to clinical contexts. Our study confirmed the general correlation between morphotype and GPL levels in smooth strains yet unveiled more variability within morphotype groups than previously recognised, particularly during intracellular infection. As the rough morphotype is of highest clinical concern, these findings contribute to the expanding knowledge base surrounding Mab infections, offering insights that can steer diagnostic methodologies, and treatment approaches.