Hunting for a viral proxy in bioaerosols of swine buildings using molecular detection and metagenomics.

There are limited biosecurity measures directed at preventing airborne transmission of viruses in swine. The effectiveness of dust mitigation strategies such as oil sprinkling, to decrease risk of airborne virus transmission are unknown. Metagenomics and qPCR for common fecal viruses were used to hunt for a ubiquitous virus to serve as a proxy when evaluating the efficiency of mitigation strategies against airborne viral infectious agents. Air particles were collected from swine buildings using high-volume air samplers. Extracted DNA and RNA were used to perform specific RT-qPCR and qPCR and analyzed by high-throughput sequencing. Porcine astroviruses group 2 were common (from 102 to 105 genomic copies per cubic meter of air or gc/m3, 93% positivity) while no norovirus genogroup II was recovered from air samples. Porcine torque teno sus virus were detected by qPCR in low concentrations (from 101 to 102 gc/m3, 47% positivity). Among the identified viral families by metagenomics analysis, Herelleviridae, Microviridae, Myoviridae, Podoviridae, and Siphoviridae were dominant. The phage vB_AviM_AVP of Aerococcus was present in all air samples and a newly designed qPCR revealed between 101 and 105 gc/m3 among the samples taken for the present study (97% positivity) and banked samples from 5- and 15-year old studies (89% positivity). According to the present study, both the porcine astrovirus group 2 and the phage vB_AviM_AVP of Aerococcus could be proxy for airborne viruses of swine buildings.

The occurrence of Aerococcus urinaeequi and non-aureus staphylococci in raw milk negatively correlates with Escherichia coli clinical mastitis.

Escherichia coli is a common environmental pathogen associated with clinical mastitis (CM) in dairy cattle. There is an interest in optimizing the udder microbiome to increase the resistance of dairy cattle to E. coli CM; however, the details of which members of the healthy udder microbiome may play a role in antagonizing E. coli are unknown. In this study, we characterized the bacterial community composition in raw milk collected from quarters of lactating Holstein dairy cows that developed E. coli CM during lactation, including milk from both healthy and diseased quarters (n = 1,172). The milk microbiome from infected quarters was compared before, during, and after CM. A combination of 16S rRNA gene amplicon and metagenomic sequencing was used generate data sets with a high level of both depth and breadth. The microbial diversity present in raw milk significantly decreased in quarters experiencing E. coli CM, indicating that E. coli displaces other members of the microbiome. However, the diversity recovered very rapidly after infection. Two genera, Staphylococcus and Aerococcus, and the family Oscillospiraceae were significantly more abundant in healthy quarters with low inflammation. Species of these genera, Staphylococcus auricularis, Staphylococcus haemolyticus, and Aerocussus urinaeequi, were identified by metagenomics. Thus, these species are of interest for optimizing the microbiome to discourage E. coli colonization without triggering inflammation.IMPORTANCEIn this study, we show that E. coli outcompetes and displaces several members of the udder microbiome during CM, but that microbial diversity recovers post-infection. In milk from quarters which remained healthy, the community composition was often highly dominated by S. auricularis, S. haemolyticus, A. urinaeequi, and S. marcescens without increases in somatic cell count (SCC). Community dominance by these organisms, without inflammation, could indicate that these species might have potential as prophylactic probiotics which could contribute to colonization resistance and prevent future instances of E. coli CM.

Emerging pathogens Aerococcus urinae and Aerococcus sanguinicola from a Canadian tertiary care hospital.

Background: Aerococcus urinae and Aerococcus sanguinicola are emerging pathogens linked with urinary tract infections. We present a case series of A. urinae and A. sanguinicola isolates characterizing the spectrum of clinical presentation, microbiological characteristics and antimicrobial sensitivities. Methods: Retrospective chart review was performed on patients who grew positive cultures for A. urinae and A. sanguinicola identified on MALDI-TOF in Saskatchewan from January to June 2023. Demographic and clinical variables, antimicrobial susceptibility and prescription were documented. Results: This cohort (n = 115) had a median age 82 years. A. urinae and A. sanguinicola infections spanned from urinary tract infection (n = 96) to urosepsis (n = 6). These infections were predominantly monomicrobial (73.9%) and were susceptible to ceftriaxone, penicillin G and vancomycin. Antimicrobials were seldom prescribed within the urinary tract infection cohort (31.2%). Conclusion: Untreated A. urinae and A. sanguinicola infections can precipitate into urosepsis. The reported antimicrobial susceptibility for these Aerococcus isolates should be utilized to provide appropriate antimicrobial coverage.

Aerococcus urinae and Aerococcus sanguinicola are bacteria that can cause urine infections. They are often overlooked and thought to be unable to cause serious blood infections, such as sepsis. We collected data on 87 cases of A. urinae and 28 cases of A. sanguinicola to show that these bacteria can cause urine and blood infections in elderly patients. We also looked at other studies and summarized that patients with serious blood infections from these bacteria often had a previous urine infection from these same bacteria. These bacteria can be resistant to a common antibiotic used to treat urine infections. It is important to test and report if these bacteria are resistant to this common antibiotic and doctors must be aware that they can cause serious blood infections if not treated with the correct antibiotics.

Evaluation of gradient diffusion in vitro susceptibility testing of Aerococcus urinae.

OBJECTIVE: Aerococcus urinae antimicrobial susceptibility testing can be performed via broth microdilution with Mueller-Hinton broth supplemented with lysed horse blood. We sought to compare this with the commonly used gradient diffusion method. METHODS: We compared broth microdilution with Mueller-Hinton broth supplemented with lysed horse blood and gradient diffusion via Mueller-Hinton agar supplemented with sheep blood for 190 A. urinae isolates against 16 antimicrobials. RESULTS: No antimicrobials demonstrated more than 90% essential and categorical agreement, and fewer than 3% demonstrated major and very major error rates. Trimethoprim-sulfamethoxazole demonstrated an 81% major error rate and ceftriaxone demonstrated a 76% very major error rate. Agar dilution with lysed horse blood was performed for trimethoprim-sulfamethoxazole against 94 isolates and showed 100% susceptibility, consistent with previous studies. CONCLUSIONS: Given its limitations in detecting resistant strains, our findings cannot support the routine use of gradient diffusion with Mueller-Hinton agar supplemented with sheep blood for A. urinae in lieu of the Clinical and Laboratory Standards Institute method. Our results suggest that A. urinae is usually susceptible to penicillin, linezolid, tetracycline, and vancomycin. Future studies should evaluate alternative testing methods for clinical microbiology laboratories.

Aerococcus urinae endocarditis - A case report.

Aerococcus urinae is a gram-positive coccus bacterium with a previously underestimated prevalence due to morphological similarities to other gram-positive cocci. Development of newer diagnostic technologies (such as matrix-assisted laser desorption ionization time-of-flight mass spectrometry MALDI-TOF) led to increased recognition of Aerococcus urinae as causative organism mainly for urinary tract infections. Its antibiotic susceptibility poses some challenges, with resistance to some drugs of choice for urinary tract infection. We report a case of a 69-year-old male with infective endocarditis of the mitral valve, who initially presented with fever and shoulder pain to the emergency department. The patient reported an episode of obstructive renal infection two weeks earlier, which was treated with trimethoprim-sulfamethoxazole. The unusual presentation with shoulder pain and a new heart murmur led to suspicion of endocarditis. Urine and blood cultures were positive for Aerococcus urinae, echocardiography revealed vegetations on the mitral valve with severe mitral insufficiency. After two weeks of antibiotic treatment, mitral valve replacement was performed, from which the patient recovered. Reports of Aerococcus urinae endocarditis are still limited in number. On the other side, Aerococcus urinae is an emerging bacterial uropathogen with greater relevance than previously believed. We review the case reports of Aerococcus urinae endocarditis and newest literature about its presentation, course, and clinical management.

Biological characterization of the phage lysin AVPL and its efficiency against Aerococcus viridans-induced mastitis in a murine model.

Aerococcus viridans (A. viridans) is an important opportunistic zoonotic pathogen that poses a potential threat to the animal husbandry industry, such as cow mastitis, due to the widespread development of multidrug-resistant strains. Phage lysins have emerged as a promising alternative antibiotic treatment strategy. However, no lysins have been reported to treat A. viridans infections. In this study, the critical active domain and key active sites of the first A. viridans phage lysin AVPL were revealed. AVPL consists of an N-terminal N-acetylmuramoyl-L-alanine amidase catalytic domain and a C-terminal binding domain comprising two conserved LysM. H40, N44, E52, W68, H147, T157, F60, F64, I77, N92, Q97, H159, V160, D161, and S42 were identified as key sites for maintaining the activity of the catalytic domain. The LysM motif plays a crucial role in binding AVPL to bacterial cell wall peptidoglycan. AVPL maintains stable activity in the temperature range of 4-45°C and pH range of 4-10, and its activity is independent of the presence of metal ions. In vitro, the bactericidal effect of AVPL showed efficient bactericidal activity in milk samples, with 2 µg/mL of AVPL reducing A. viridans by approximately 2 Log10 in 1 h. Furthermore, a single dose (25 µg) of lysin AVPL significantly reduces bacterial load (approximately 2 Log10) in the mammary gland of mice, improves mastitis pathology, and reduces the concentration of inflammatory cytokines (TNF-α, IL-1ß, and IL-6) in mammary tissue. Overall, this work provides a novel alternative therapeutic drug for mastitis induced by multidrug-resistant A. viridans. IMPORTANCE: A. viridans is a zoonotic pathogen known to cause various diseases, including mastitis in dairy cows. In recent years, there has been an increase in antibiotic-resistant or multidrug-resistant strains of this pathogen. Phage lysins are an effective approach to treating infections caused by multidrug-resistant strains. This study revealed the biological properties and key active sites of the first A. viridans phage lysin named AVPL. AVPL can effectively kill multidrug-resistant A. viridans in pasteurized whole milk. Importantly, 25 µg AVPL significantly alleviates the symptoms of mouse mastitis induced by A. viridans. Overall, our results demonstrate the potential of lysin AVPL as an antimicrobial agent for the treatment of mastitis caused by A. viridans.

Characteristics of urinary tract infections in children with positive urine culture for Aerococcusurinae.

Urinary tract infections caused by Aerococcus urinae have rarely been reported in children, and the clinical characteristics remain unclear. We reviewed medical records of children whose urine cultures grew A. urinae (≥104 CFU/mL) at a tertiary children's hospital in Tokyo, Japan. We found 17 pediatric patients in a review of 22,769 urine cultures between June 2006 and May 2022. The median age of 17 patients was 10.7 years (IQR 8-13 years), and males represented 76.5 % of the patients. Sixteen patients (94.1 %) had underlying urological conditions (neurogenic bladder, vesicoureteral reflux, urethral stenosis, bladder exstrophy, or urinary catheterization). The chief symptoms were fever (35.3 %), malodorous urine (23.5 %), nausea (11.8 %), and back pain (5.9 %). Ten patients were asymptomatic. Pyelonephritis was diagnosed in five male patients. All of them had underlying abnormal conditions of the bladder, and two had malodorous urine. All patients had favorable outcomes after 10-14 days of ampicillin/amoxicillin-based antimicrobial therapy.

Macrolide Resistance in the Aerococcus urinae Complex: Implications for Integrative and Conjugative Elements.

The recognition of the Aerococcus urinae complex (AUC) as an emerging uropathogen has led to growing concerns due to a limited understanding of its disease spectrum and antibiotic resistance profiles. Here, we investigated the prevalence of macrolide resistance within urinary AUC isolates, shedding light on potential genetic mechanisms. Phenotypic testing revealed a high rate of macrolide resistance: 45%, among a total of 189 urinary AUC isolates. Genomic analysis identified integrative and conjugative elements (ICEs) as carriers of the macrolide resistance gene ermA, suggesting horizontal gene transfer as a mechanism of resistance. Furthermore, comparison with publicly available genomes of related pathogens revealed high ICE sequence homogeneity, highlighting the potential for cross-species dissemination of resistance determinants. Understanding mechanisms of resistance is crucial for developing effective surveillance strategies and improving antibiotic use. Furthermore, the findings underscore the importance of considering the broader ecological context of resistance dissemination, emphasizing the need for community-level surveillance to combat the spread of antibiotic resistance within the urinary microbiome.

Complete genome sequences of Aerococcus loyolae ATCC TSD-300T, Aerococcus mictus ATCC TSD-301T, and Aerococcus tenax ATCC TSD-302T.

Previously identified under the single designation of Aerococcus urinae, three distinct taxonomic species have been distinguished as Aerococcus loyolae, Aerococcus mictus, and Aerococcus tenax. Here, we present the complete genome sequences of the type strains of these species assembled via a combination of short-read and long-read sequencing techniques.Registered at ClinicalTrials.gov (NCT01166438).

Aerococcus Urinae Infection Causing Malodorous Urine in a Child: A Case Report.

Aerococcus urinae (A. urinae) infection, primarily observed in elderly patients, is a rare yet emerging occurrence in the pediatric population. Advances in laboratory techniques have facilitated the increased identification of these bacteria in human infections. There have been only a few recent cases reported among children. The scarcity of literature on the clinical presentation and management of such infections in children presents a challenge for pediatricians. Here, we present the case of a 15-month-old male child with Down syndrome who presented with malodorous urine but lacked other typical symptoms of urinary tract infection. Upon investigation, urine analysis revealed pyuria, and urine culture confirmed A. urinae infection. The patient also exhibited underlying bilateral mild to moderate hydronephrosis. Successful treatment was achieved with a three-day course of amoxicillin, leading to symptom resolution. This case underscores the significance of promptly identifying A. urinae infection in pediatric patients presenting with malodorous urine, as a timely intervention with a short course of treatment may avert more severe and invasive infections.

Aerococcus kribbianus sp. nov., a facultatively anaerobic bacterium isolated from pig faeces.

Two cocci-shaped, facultatively anaerobic, Gram-positive bacteria isolated from the faeces of a pig were designated as strains YH-aer221T and YH-aer222. Analysis of the 16S rRNA gene sequences revealed that the isolates were most closely related to Aerococcus suis JCM 18035T with 96.6 % similarity. The multi-locus sequence tree revealed that the isolates formed a sub-cluster adjacent to A. suis JCM 18035T. The average nucleotide identity values for the isolates and their most closely related strains were 71.8 and 71.7 %, respectively; and the digital DNA-DNA hybridization values for the isolates and their most closely related strains were 25.6 and 25.5 %, respectively. The main fatty acids were C18 : 1ω9c, C16 : 0 and C18 : 0. The cell wall contained the meso-diaminopimelic acid-based peptidoglycan. The two isolates shared the same metabolic pathways. Isolates YH-aer221T and YH-aer222 harboured the same CRISPR array with 33 and 46 spacers, respectively. Single-genome vs. metagenome analysis showed that the genomes of the isolates were not found in the available metagenome database. Given their chemotaxonomic, phenotypic and phylogenetic properties, YH-aer221T (= KCTC 25571T=JCM 35699T) and YH-aer222 (=KCTC 25573=JCM 35700) represent a novel taxon. The name Aerococcus kribbianus sp. nov. is proposed.

A case report of melanosis of the bladder within a bladder diverticulum and associated Aerococcus urinary infection.

Background: Melanosis vesicae is a rare condition characterized by the deposition of melanin within the bladder urothelium. Case presentation: We present a case of a 72-year-old male with a history of recurrent urinary retention, bladder diverticula, and concurrent Aerococcus urinary tract infection who presented with left-sided abdominal pain. Cystoscopy revealed diffuse black splotch lesions throughout the bladder and two diverticula. Histopathological examination confirmed the diagnosis of melanosis vesicae. The patient ultimately underwent an open bladder diverticulectomy. Conclusion: The potential associations between melanosis vesicae, urinary tract malignancies and concurrent conditions such as bladder diverticula and urinary infections warrant further investigation.

Aerococcus viridans Phage Lysin AVPL Had Lytic Activity against Streptococcus suis in a Mouse Bacteremia Model.

Streptococcus suis (S. suis) is a swine pathogen that can cause sepsis, meningitis, endocarditis, and other infectious diseases; it is also a zoonotic pathogen that has caused a global surge in fatal human infections. The widespread prevalence of multidrug-resistant S. suis strains and the decline in novel antibiotic candidates have necessitated the development of alternative antimicrobial agents. In this study, AVPL, the Aerococcus viridans (A. viridans) phage lysin, was found to exhibit efficient bactericidal activity and broad lytic activity against multiple serotypes of S. suis. A final concentration of 300 µg/mL AVPL reduced S. suis counts by 4-4.5 log10 within 1 h in vitro. Importantly, AVPL effectively inhibited 48 h S. suis biofilm formation and disrupted preformed biofilms. In a mouse model, 300 µg/mouse AVPL protected 100% of mice from infection following the administration of lethal doses of multidrug-resistant S. suis type 2 (SS2) strain SC19, reduced the bacterial load in different organs, and effectively alleviated inflammation and histopathological damage in infected mice. These data suggest that AVPL is a valuable candidate antimicrobial agent for treating S. suis infections.

Aerococcus agrisoli sp. nov., isolated from paddy soil.

A non-spore-forming, Gram-stain-positive, short rod-shaped strain, designated SJQ22T, was isolated from a paddy soil sample collected in Shanghai, PR China. A comparative analysis of 16S rRNA gene sequences showed that strain SJQ22T fell within the genus Aerococcus, forming a clear cluster with the type strains of Aerococcus viridans (98.6 % sequence similarity) and Aerococcus urinaeequi (98.5 % sequence similarity). Strain SJQ22T grew at 30-45 °C (optimum, 30 °C), pH 6.0-8.0 (optimum, pH 7.0) and with a NaCl concentration of 0-4 % (optimum, 1 %). Cells were negative for oxidase and catalase activity. Chemotaxonomic analysis showed that strain SJQ22T possessed C16:0 and C18:1 ω9c as the predominant fatty acids. The DNA G + C content was 39.0 mol%. Strain SJQ22T exhibited DNA-DNA relatedness levels of 13±2 % with A. viridans ATCC 11563T and 9±2 % with A. urinaeequi IFO 12173T. Based on the data obtained, strain SJQ22T represents a novel species of the genus Aerococcus, for which the name Aerococcus agrisoli sp. nov. is proposed. The type strain is SJQ22T (=JCM 33111T=CCTCC AB 2018283T).

Aerococcus urinae infection of the pubic symphysis: A rare case report.

Aerococcus urinae is a Gram-positive, catalase-negative coccus, rarely responsible for urinary tract infections and seldom described for musculoskeletal infections like spondylodiscitis. An 86-year-old man presented to our hospital for groin pain without fever. Pelvic CT-guided biopsy revealed an A. urinae pubic symphysis osteomyelitis. He received a treatment by amoxicillin per os for six weeks, and did not need any surgery. An eight -month- follow-up showed a favorable evolution. Pubic symphysis infection can be induced by a wide variety of pathogens, and may have very different clinical presentations. Some authors recommend systematic surgery, but in case of susceptible pathogen associated with a low level of joint destruction, medical treatment alone should be sufficient to cure and make surgery unnecessary.

Taxonomic considerations on Aerococcus urinae with proposal of subdivision into Aerococcus urinae, Aerococcus tenax sp. nov., Aerococcus mictus sp. nov., and Aerococcus loyolae sp. nov.

Average nucleotide identity analysis, based on whole genome sequences of 115 strains previously identified as Aerococcus urinae, an emerging uropathogen, discriminates at least six unique genomic taxa. The whole genome analysis affords clearer species boundaries over 16S rRNA gene sequencing and traditional phenotypic approaches for the identification and phylogenetic organization of Aerococcus species. The newly described species can be differentiated by matrix-assisted laser desorption ionization time-of-flight analysis of protein signatures. We propose the emendation of the description of A. urinae (type strain ATCC 51268T = CCUG 34223T=NCFB 2893) and the names of Aerococcus tenax sp. nov. (ATCC TSD-302T = DSM 115700T = CCUG 76531T=NR-58630T), Aerococcus mictus sp. nov. (ATCC TSD-301T = DSM 115699T = CCUG 76532T=NR-58629T), and Aerococcus loyolae sp. nov. (ATCC TSD-300T = DSM 115698T = CCUG 76533T=NR-58628T) for three of the newly identified genomic taxa.

Bacteriophage Therapy to Control Bovine Mastitis: A Review.

Bovine mastitis is a polymicrobial disease characterised by inflammation of the udders of dairy and beef cattle. The infection has huge implications to health and welfare of animals, impacting milk and beef production and costing up to EUR 32 billion annually to the dairy industry, globally. Bacterial communities associated with the disease include representative species from Staphylococcus, Streptococcus, Enterococcus, Actinomyces, Aerococcus, Escherichia, Klebsiella and Proteus. Conventional treatment relies on antibiotics, but antimicrobial resistance, declining antibiotic innovations and biofilm production negatively impact therapeutic efficacy. Bacteriophages (phages) are viruses which effectively target and lyse bacteria with extreme specificity and can be a valuable supplement or replacement to antibiotics for bovine mastitis. In this review, we provide an overview of the etiology of bovine mastitis, the advantages of phage therapy over chemical antibiotics for the strains and research work conducted in the area in various model systems to support phage deployment in the dairy industry. We emphasise work on phage isolation procedures from samples obtained from mastitic and non-mastitic sources, characterisation and efficacy testing of single and multiple phages as standalone treatments or adjuncts to probiotics in various in vitro, ex vivo and in vivo bovine mastitis infection models. Furthermore, we highlight the areas where improvements can be made with focus on phage cocktail optimisation, formulation, and genetic engineering to improve delivery, stability, efficacy, and safety in cattle. Phage therapy is becoming more attractive in clinical medicine and agriculture and thus, could mitigate the impending catastrophe of antimicrobial resistance in the dairy sector.

Epidemiology and urological pathogenic potential of Aerococcus species in greater Glasgow and Clyde (Descriptive study of Aerococcus urinae in blood culture and urinary samples: clinical importance and potential marker of urinary tract pathology).

Introduction. Aerococcus species in particular A. urinae are increasingly reported as causative agents of bacteraemia, urinary tract infection, sepsis, and endocarditis. We sought to establish the epidemiology of A. urinae in Glasgow hospitals and whether the presence of the organism in clinical isolates could be an indicator of undiagnosed urinary tract pathology.Hypothesis/Gap statement. The knowledge gap among clinical staffs on Aerococcus species as emerging pathogens can be filled by understanding its epidemiology and clinical importance.Aim. Describe the epidemiology and clinical importance of Aerococcus urinae.Methodology. We reviewed positive blood cultures with Aerococcus species (2017-2021) and urinary isolates (2021) in Glasgow hospitals. Data were collected from clinical and laboratory database systems.Results. All 22 positive blood cultures were A. urinae and sensitive to amoxicillin, vancomycin, and ciprofloxacin. The median age was 80.5; the majority was male (18). In total, 15/22 (68 %) were diagnosed with urinary tract infection. Thirteen were treated with amoxicillin. No cases of infective endocarditis were noted. One patient was subsequently diagnosed with bladder carcinoma. All 83 positive urinary isolates in 72 patients were A. urinae. One was resistant to amoxicillin; two to ciprofloxacin; all sensitive to nitrofurantoin and vancomycin. The majority was female (43/83), the median age was 80. The commonest risk factors were underlying malignancy including bladder cancer (5/18), chronic kidney disease (17) and diabetes (16). Clinical data was unavailable in 24 episodes. Of these, 41/59 (69.5 %) were diagnosed with urinary tract infection. One patient was subsequently diagnosed with metastatic renal cancer while bladder wall lesions were identified in three patients, two of whom were waiting for an urology review at the time of study. Thirteen patients (18 %) had 1 year recurrent bacteriuria and three were not treated on initial episode.Conclusion. A. urinae are emerging pathogens and are likely to become more common due to advances in laboratory technologies and an ageing population. Clinical teams should be aware of their urological pathogenic potential and not dismiss them as contaminants. Whether Aerococcus infection is a potential indicator for undiagnosed urinary tract malignancy warrants further studies.

Aero to the heart: A case of aortic valve endocarditis caused by Aerococcus Urinae in an elderly woman.

Aerococcus urinae (A. urinae) is an infrequent cause of infective endocarditis (IE) and few cases have been reported especially in older women. As of this publication, there are 31 reported cases of IE caused by aerococcus urinae, and of these, 4 are of women, 3 of which are aged > 75 years. Here, we describe a case of A. urinae endocarditis in an 80-year-old woman presenting with worsening fatigue. A diagnosis of native aortic valve endocarditis was established based on characteristic findings of aortic valvular vegetation on transesophageal echocardiogram along with isolation of A. urinae on blood cultures.