First isolation and whole-genome sequencing of Trueperella abortisuis from a goat in Canada.

Objective: The present study reports the first isolation and whole-genome sequencing of a Trueperella abortisuis bacterium from a goat. Animals and sample: The T. abortisuis was isolated from the uterus of a goat following an abortion. Procedure: The T. abortisuis was identified by pure culture phenotype and MALDI-TOF analysis and further characterized by whole-genome sequencing. Results: This isolate was reliably identified as T. abortisuis and showed similar properties to type strain T. abortisuis DSM 19515T, which was recovered from a sow following an abortion. The assembled genome of this isolate was 2 564 866 bp long with a GC content of 63.9%. A total of 30 virulence-related genes were determined, suggesting the pathogenic potential of this organism. Conclusion and clinical relevance: This study details the first isolation of T. abortisuis from goats. The genotypic findings of this isolate will serve as a baseline description for any similar future studies.

Premier isolement et séquençage du génome entier de Trueperella abortisuis provenant d'une chèvre au Canada. Objectif: La présente étude rapporte le premier isolement et séquençage du génome entier d'un isolat de Trueperella abortisuis provenant d'une chèvre. Animaux et échantillon: Le T. abortisuis a été isolé de l'utérus d'une chèvre à la suite d'un avortement. Procédure: Le T. abortisuis a été identifié par un phénotype de culture pure et analyse par MALDI-TOF, puis caractérisé par séquençage du génome entier. Résultats: Cet isolat a été identifié de manière fiable comme étant T. abortisuis et a montré des propriétés similaires à la souche type T. abortisuis DSM 19515T, qui a été récupérée chez une truie après un avortement. Le génome assemblé de cet isolat mesurait 2 564 866 pb avec une teneur en GC de 63,9 %. Au total, 30 gènes liés à la virulence ont été déterminés, suggérant le potentiel pathogène de cet organisme. Conclusion et pertinence clinique: Cette étude détaille le premier isolement de T. abortisuis chez la chèvre. Les résultats génotypiques de cet isolat serviront de description de base pour toute étude future similaire.(Traduit par Dr Serge Messier).

Whole genome-based antimicrobial resistance, virulence, and phylogenetic characteristics of Trueperella pyogenes clinical isolates from humans and animals.

Trueperella pyogenes is an opportunistic zoonotic bacterial pathogen, whose antimicrobial resistance, virulence, and genetic relatedness between strains from animals and humans are barely studied. These characteristics were therefore analyzed for clinical T. pyogenes strains from 31 animals of 11 different species and 8 humans determining their complete circular genome sequence and antimicrobial susceptibility. The MICs of 19 antimicrobials including 3 antiseptics correlated to the resistance genes identified in silico within the genomes revealing a predominance of resistance to streptomycin (aadA9), sulfamethoxazole (sul1), and tetracycline (tet(33), tet(W/N/W)) among strains from humans and cattle. Additional resistance genes (erm(X), erm(56), cmx, drfA1, aadA1, aph(3'')-Ib (strA), aph(6)-Id (strB), aac(3)-IVa, aph(4)-Ia) were found only sporadically. The resistance genes were localized on genetic elements integrated into the chromosome. A cgMLST-based phylogenetic analysis revealed two major clusters each containing genetically diverse strains. The human strains showed the closest relatedness to strains from cattle. Virulence genes coding for fimbriae (fimA, fimC), neuroamidase (nanP, nanH), pyolysin (plo), and collagen binding protein (cbpA) were identified in strains from different hosts, but no correlation was observed between virulence factors and strain origin. The existence of resistance genes typically found in Gram-negative bacteria within the Gram-positive T. pyogenes indicates a wider capacity to adapt to antimicrobial selective pressure. Moreover, the presence of similar antimicrobial resistance profiles found in cattle and human strains as well as their closest relatedness suggests common zoonotic features and cattle as the potential source for human infections.

Antimicrobial efficiency of bromhexine hydrochloride against endometritis-causing Escherichia coli and Trueperella pyogenes in bovines.

In this study, the main agents associated with endometritis in cows in the state of Santa Catarina, Brazil, were identified and the resistance profile and virulence mechanisms of the bacterial isolates were evaluated. Isolates of Escherichia coli and Trueperella pyogenes were tested for their biofilm forming ability and the antimicrobial action of bromhexine hydrochloride in combination with other antimicrobials. A total of 37 uterine lavage samples were collected from cows with endometritis. Of the 55 bacteria isolated, 25.4% were identified as T. pyogenes and 16.3% as E. coli. The bacterial isolates showed greater resistance to sulfamethoxazole + trimethoprim (58.2%) and tetracycline (56.3%). Among the species, E. coli showed the highest resistance rates, with 100% of isolates showing resistance to amoxicillin, streptomycin, and gentamicin. The results of the minimum inhibitory concentration for the T. pyogenes isolates showed that 91.6% of the isolates were resistant to enrofloxacin and tetracycline, and 75% were resistant to ceftiofur and sulfamethoxazole + trimethoprim. All E. coli and T. pyogenes isolates showed biofilm forming ability. The plo, fimA, and nanH genes were identified in 100% of T. pyogenes isolates. In parallel, 100% of E. coli isolates had the fimH gene, and 11.1% had the csgD gene. Bromhexine hydrochloride showed antimicrobial activity against 100% of E. coli isolates and 66.6% of T. pyogenes isolates. Furthermore, when associated with antimicrobials, bromhexine hydrochloride has a synergistic and additive effect, proving to be an option in the treatment of endometritis in cows and an alternative for reducing the use of antimicrobials.

Multiphasic investigations imply transfer of orange-/red-pigmented strains of the bean pathogen Curtobacterium flaccumfaciens pv. flaccumfaciens to a new species as C. aurantiacum sp. nov., elevation of the poinsettia pathogen C. flaccumfaciens pv. poinsettiae to the species level as C. Poinsettiae sp. nov., and synonymy of C. albidum with C. citreum.

Curtobacterium flaccumfaciens (Microbacteriaceae), a plant-pathogenic coryneform species includes five pathovars with valid names and a number of proposed - but unvalidated - new members. In this study, phenotypic features and DNA similarity indexes were investigated among all C. flaccumfaciens members. Results showed that the C. flaccumfaciens pv. poinsettiae strains causing bacterial canker of Euphorbia pulcherrima in the USA as well as the orange-/red-pigmented strains of C. flaccumfaciens pv. flaccumfaciens pathogenic on dry beans in Iran are too distinct from each other and from the type strain of the species to be considered members of C. flaccumfaciens. Hence, the latter two groups were elevated at the species level as C. poinsettiae sp. nov. (ATCC 9682T = CFBP 2403T = ICMP 2566T = LMG 3715T = NCPPB 854T as type strain), and C. aurantiacum sp. nov. (50RT = CFBP 8819T = ICMP 22071T as type strain). Within the emended species C. flaccumfaciens comb. nov., yellow-pigmented strains causing bacterial wilt of dry beans and those causing bacterial canker of Euphorbia pulcherrima in Europe were retained as C. flaccumfaciens pv. flaccumfaciens and C. flaccumfaciens pv. poinsettiae, respectively; while taxonomic position of the sugar beet pathogen C. flaccumfaciens pv. beticola ATCC BAA144PT was confirmed. The newly described onion pathogen C. allii was also reclassified as C. flaccumfaciens pv. allii with the pathotype strain LMG 32517PT. Furthermore, C. flaccumfaciens pv. basellae causing bacterial leaf spot of malabar spinach (Basella rubra) was transferred to C. citreum pv. basellae with ATCC BAA143PT as pathotype.

A rare case of male Fournier's gangrene with mixed Actinomyces turicensis infection.

BACKGROUND: Fournier's gangrene (FG), a urological emergency with high mortality, is an infectious necrotizing fasciitis of the perineal and genital regions. The majority of FG is caused by polymicrobial organisms involving mixed aerobes and anaerobes but rarely reveals Actinomyces species. CASE PRESENTATION: We report a healthy 67-year-old Asian male who presented with rapidly progressive painful swelling of the scrotum. Clinically diagnosed with FG, the patient underwent an emergency radical debridement, followed by broad-spectrum antibiotics and negative pressure wound therapy. The identification of the causative microorganisms showed Actinomyces turicensis and the antibiotic treatment was adjusted accordingly. After wound bed preparation, we took split-thickness skin grafts to cover the scrotal wound. Active management to minimize faecal contamination was applied throughout the whole course of treatment and repair. The patient was satisfied with the outcome. This was an extremely rare case of A. turicensis as the main causative pathogen of FG. CONCLUSIONS: FG due to Actinomyces species is rarely reported, but we should still consider this pathogenic microorganism that has long been neglected.

Helcococcus ovis associated with septic arthritis and bursitis in calves - a case report.

BACKGROUND: Septic arthritis often occurs in young calves when the passive transfer of maternal immunoglobulins has failed, which results in hypogammaglobulinaemia in the calf. Another important cause is suboptimal herd health management which often leads to general health impairment and, subsequently, to septic arthritis. CASE PRESENTATION: A dairy farmer consulted the Herd Health Service of the University Clinic reporting general herd health impairment, a high incidence of respiratory diseases, unsatisfactory weight gain and arthritis in calves, as well as mastitis and high milk cell counts. Clinical examinations were performed, and diagnostic measures were taken. A transtracheal lavage (TTL) was performed, and synovial swab samples were taken from the carpal joint and the subcutaneous tarsal bursae of two calves. Microbiological examinations of synovial swabs revealed co-infections of Trueperella pyogenes and Helcococcus ovis in one calf and Helcococcus ovis in pure culture in the other. The TTLs confirmed the presence of Mycoplasma spp. associated with respiratory diseases. CONCLUSIONS: Helcococcus ovis is currently regarded as a co-infective bacterial agent. However, it seems to play a significant role as the primary pathogen in this case.

An Uncommon Case of Trueperella pyogenes Infection in an Adult Backyard Rooster and a Retrospective Study; 2000-20.

Trueperella pyogenes is an opportunistic Gram-positive bacterium that induces purulent lesions and abscesses in cattle, small ruminants, and swine. In birds, T. pyogenes infections have been linked to lameness and osteomyelitis in turkeys (Phasianidae) and hepatic fibriscess in turkeys and pigeons (Columbidae). An 18-mo-old backyard rooster with a history of progressive emaciation was submitted to the California Animal Health and Food Safety (CAHFS) laboratory system. At necropsy, unusual numerous miliary granulomas were identified, primarily in the spleen, but granulomas were also observed in air sacs and lungs. Microscopically, few to moderate numbers of granulomas with giant cells were observed in the spleen, lung, air sacs, and crop composed of necrosis and mixed inflammatory cell inflammation including multinucleated giant cells, fibrin deposition, and fibrosis. Trueperella pyogenes was isolated from the air sacs and trachea. Avibacterium paragallinarum PCR was positive from the tracheal swab. A retrospective analysis of CAHFS data on T. pyogenes between 2000 and 2020 identified 24 cases in avian species: chickens (Gallus gallus domesticus; 16/24), turkeys (5/24), Pekin duck (Anas platyrhynchos domesticus; 1/24), parrot (Psittaciformes; 1/24), and pheasant (Phasianidae; 1/24). Although T. pyogenes infection in birds is rare, the clinical signs and gross lesions might be indistinguishable from avian mycobacteriosis in some cases and should be considered in the differential diagnosis.

Reporte de caso­Un caso no común de infección por Trueperella pyogenes en un gallo adulto de traspatio y un estudio retrospectivo; entre los años 2000-20. Trueperella pyogenes es una bacteria grampositiva oportunista que induce lesiones purulentas y abscesos en bovinos, pequeños rumiantes y porcinos. En las aves, las infecciones por T. pyogenes se han relacionado con cojera y osteomielitis en pavos (Phasianidae) y fibrosis hepática en pavos y palomas (Columbidae). Un gallo de traspatio de 18 meses de edad con antecedentes de emaciación progresiva fue enviado al sistema de Laboratorios de Salud Animal y Seguridad Alimentaria de California (CAHFS). En la necropsia, se identificaron numerosos granulomas miliares inusuales, principalmente en el bazo, pero también se observaron granulomas en los sacos aéreos y los pulmones. Microscópicamente, se observaron pocos a moderados granulomas con células gigantes en el bazo, pulmón, sacos aéreos y buche compuesto por necrosis e inflamación celular inflamatoria mixta, incluidas células gigantes multinucleadas, depósito de fibrina y fibrosis. Trueperella pyogenes se aisló de los sacos aéreos y la tráquea. Un método de PCR para Avibacterium paragallinarum fue positivo realizado a partir de hisopos traqueales. Un análisis retrospectivo de los datos de CAHFS sobre T. pyogenes entre los años 2000 y 2020 identificó 24 casos en especies aviares: pollos (Gallus gallus domesticus; 16/24), pavos (5/24), pato Pekín (Anas platyrhynchos domesticus; 1/24), loro (Psittaciformes; 1/24) y faisán (Phasianidae; 1/24). Aunque la infección por T. pyogenes en aves es poco común, los signos clínicos y las lesiones macroscópicas pueden ser indistinguibles de micobacteriosis aviar en algunos casos y debe considerarse como diagnóstico diferencial.

Nanchangia anserum gen. nov., sp. nov., isolated from feces of greater white-fronted geese (Anser albifrons).

Two rod-shaped and Gram-stain-positive bacteria (strains C64T and C62) were isolated in 2020 from faeces of greater white-fronted geese (Anser albifrons) from Poyang Lake, PR China. Their optimal growth conditions were at 37 °C, pH 7.0 and with 0.5 % (w/v) NaCl. The two isolates showed a highest 16S rRNA gene sequence similarity to Bowdeniella nasicola DSM 19116T (92.1 %). Phylogenetic/phylogenomic analyses indicated that strains C64T and C62 clustered independently in the vicinity of the genera Varibaculum, Winkia and Mobiluncus within the family Actinomycetaceae, but could not be classified clearly as members of any of these known genera. The average amino acid identity values between our isolates and available genomes of members of the family Actinomycetaceae were around the genus threshold value (45-65 %). The major cellular fatty acids of the strains were C18 : 1ω9c and C16 : 0. The predominant polar lipids were phosphatidylinositol, phosphatidylglycerol, phosphatidylcholine, diacylglycerol, triacylglycerol and cardiolipin. The amino acid composition of peptidoglycan contained alanine, glutamic acid and glycine. The major respiratory menaquinones were MK-8(H4) and MK-9(H4). The whole cell sugars included galactose, arabinose and glucose. On the basis of the results of the 16S rRNA gene sequences comparison, whole-genome phylogenomic analysis, phenotypic and chemotaxonomic characteristics, we propose that strains C64T and C62 represent a novel species belonging to a novel genus within the family Actinomycetaceae, for which the name Nanchangia anserum gen. nov., sp. nov. is proposed. The type strain is Nanchangia anserum C64T (=CGMCC 1.18410T=GDMCC 1.1969T=KCTC 49511T=KACC 22143T).

Actinomycosis in a gray four-eyed opossum (Philander opossum) caused by a novel species of Schaalia.

BACKGROUND: Infective lesions of the jaws and adjacent tissues (lumpy jaw disease, LJD) have been recognized as one major cause of death of captive macropods. Fusobacterium necrophorum and Actinomyces species serve as the main source of LJD in kangaroos and wallabies. Currently, little is reported about LJD or similar diseases in opossums. CASE PRESENTATION: Here we report a case of actinomycosis resembling the entity lumpy jaw disease in a gray four-eyed opossum, caused by a novel species of Schaalia. A 2.8 year old male Philander opossum was presented with unilateral swelling of the right mandible. After an initial treatment with marbofloxacin, the opossum was found dead the following day and the carcass was submitted for necropsy. Postmortem examination revealed severe mandibular skin and underlying soft tissue infection with subsequent septicemia as the cause of death. Histological examination demonstrated Splendore-Hoeppli phenomenon, typically seen in classical cases of actinomycosis. Bacteriology of liver and mandibular mass yielded a previously undescribed species of Schaalia, whose 16 S rRNA gene sequence was 97.0 % identical to Schaalia canis. Whole genome sequencing of the opossum isolate and calculation of average nucleotide identity confirmed a novel species of Schaalia, for which no whole genome sequence is yet available. CONCLUSIONS: The herewith reported Schaalia infection in the gray four-eyed opossum resembling classical actinomycosis gives a novel insight into new exotic animal bacterial diseases. Schaalia species may belong to the normal oral microbiome, as in macropods, and may serve as a contributor to opportunistic infections. Due to the lack of current literature, more insights and improved knowledge about Schaalia spp. and their pathogenicity will be useful to choose appropriate therapy regimens and improve the treatment success rate and outcome in exotic and endangered species.

Septic hip abscess due to Fusobacterium nucleatum and Actinomyces turicensis in an immunocompetent SARS-CoV-2 positive patient.

A 42-year-old man was referred to the Department of Orthopedic Surgery with pain over his right greater trochanter and signs of systemic infection. CT showed an enhanced mass in his gluteus maximus as well as gas in the biceps femoris over the underlying hip joint. Tissue biopsy yielded Fusobacterium nucleatum and Actinomyces turicensis. The patient was successfully treated for 6 weeks with amoxicillin/clavulanic acid 875mg/125mg and metronidazole 500mg.

Trueperella pecoris sp. nov. isolated from bovine and porcine specimens.

A novel Gram-stain-positive bacterium was isolated from a purulent bovine milk sample, the bovine placenta from an abortion, the udder secretion of a heifer and the lung of a pig that had succumbed from suppurative bronchopneumonia in Switzerland from 2015 to 2019. The strains grew best under aerobic conditions with 5 % CO2 and colonies were non-haemolytic and greyish-white. They were non-motile and negative for catalase and oxidase. The genomes of the four strains 19M2397T, 15A0121, 15IMD0307 and 19OD0592 were obtained by sequencing. The results of phylogenetic analyses based on the 16S rRNA gene grouped them within the genus Trueperella in the family Arcanobacteriaceae. The genomes had DNA G+C contents of 61.2-62.2 mol% and showed digital DNA-DNA hybridization (dDDH) values of 21.4-22.8 % and average nucleotide identity (ANI) values of approximately 77 % to their closest relatives Trueperella pyogenes and Trueperella bernardiae. With respect to the presence in different livestock species we propose the name Trueperella pecoris sp. nov. The type strain is 19M2397T (=CCOS 1952T=DSM 111392T), isolated from the udder secretion of a heifer diagnosed with summer mastitis in 2019.

Trueperella pyogenes pharyngitis in an immunocompetent 40-year-old man.

A 40-year-old man presented to his primary care physician with a constellation of systemic symptoms and new biofilm forming along his upper airway. He had brought home a deer 10 days prior from a day of hunting, and discovered green purulent material oozing from the entrance/exit wounds. The patient smokes cigarettes and did not use any protective equipment or wash his hands between dressing the deer and smoking. Several days following exposure, he became increasingly short of breath, fatigued, constipated and developed a cough productive of orange sputum. Speaking with state wildlife biologists led to the diagnosis of a zoonotic Trueperella pyogenes infection. Initial treatment with broad spectrum antibiotics was ineffective in resolving the infection. An infectious disease appointment was made, but the patient's infection resolved with the use of a veterinarian antibiotic taken under physician's supervision.

Nitrite-producing oral microbiome in adults and children.

Recently, it was suggested that the nitrite (NO2-) produced from NO3- by oral bacteria might contribute to oral and general health. Therefore, we aimed to clarify the detailed information about the bacterial NO2-production in the oral biofilm. Dental plaque and tongue-coating samples were collected, then the NO2-producing activity was measured. Furthermore, the composition of the NO2--producing bacterial population were identified using the Griess reagent-containing agar overlay method and molecular biological method. NO2--producing activity per mg wet weight varied among individuals but was higher in dental plaque. Additionally, anaerobic bacteria exhibited higher numbers of NO2--producing bacteria, except in the adults' dental plaque. The proportion of NO2--producing bacteria also varied among individuals, but a positive correlation was found between NO2--producing activity and the number of NO2--producing bacteria, especially in dental plaque. Overall, the major NO2--producing bacteria were identified as Actinomyces, Schaalia, Veillonella and Neisseria. Furthermore, Rothia was specifically detected in the tongue coatings of children. These results suggest that dental plaque has higher NO2--producing activity and that this activity depends not on the presence of specific bacteria or the bacterial compositions, but on the number of NO2--producing bacteria, although interindividual differences were detected.

Leukocyte populations in peripheral blood of dromedary camels with clinical endometritis.

Endometritis represents the main cause of reproductive failure in dromedary camels. In dromedary camels, associations between endometritis-causing pathogen-species, disease severity, and systemic changes in the immune system have not been evaluated. In the current study, there was use of flow cytometry and immunofluorescence of membrane proteins for the evaluation of leukocyte subsets and the cellular phenotype in blood of camels with clinical endometritis and evaluations of associations with disease severity and endometritis-causing pathogens. Animals with endometritis had markedly larger numbers of total leukocytes and neutrophils. Although total lymphocyte and monocyte counts did not differ between camels with and without clinical endometritis, there were lesser numbers of total and effector CD4-positive T cells in camels with endometritis. Among monocytes, number of camel inflammatory monocytes (Mo-II) was markedly greater, whereas Mo-III numbers were less in the blood of camels with clinical endometritis. Number of inflammatory monocytes was also indicative of endometritis severity grade. Among camels with clinical endometritis, E. coli- and S. aureus-infected animals had similar endometritis grades and comparable phenotype and composition patterns of leukocytes. Neutrophils and monocytes of camels with clinical endometritis had fewer cell adhesion molecules (i.e., CD11a and CD18). Collectively, the results from the current study allowed for identification of associations between endometritis severity grade and larger numbers of inflammatory monocytes. The results also indicate there is no association between endometritis pathogen-species and changes in phenotype or composition of blood leukocytes.

Flaviflexus ciconiae sp. nov., isolated from the faeces of the oriental stork, Ciconia boyciana.

A novel Gram-stain-positive, non-motile, non-spore-forming, coccobacillus-shaped, strictly aerobic bacterium, designated strain H23T48T, was isolated from the faecal sample of an oriental stork collected from the Seoul Grand Park Zoo in Seoul, Republic of Korea. Optimal growth of strain H23T48T was observed at 30-37 °C, pH 8 and with 3 % (w/v) NaCl. 16S rRNA gene sequence-based phylogenetic analysis revealed that strain H23T48T was closely related to the genus Flaviflexus, with 97.0 and 96.7 % sequence similarities to Flaviflexus salsibiostraticola EBR4-1-2T and Flaviflexus huanghaiensis H5T, respectively. Strain H23T48T possessed MK-9(H4) as the major menaquinone and C16 : 0 (42.4 %), C18 : 1 ω9c (31.3 %) and C14 : 0 (17.7 %) as the major cellular fatty acids. The polar lipids included phosphatidylglycerol, two unidentified lipids, six unidentified phospholipids and two unidentified glycophospholipids. The amino acid composition of the cell-wall peptidoglycan was l-alanine, l-lysine, d-glutamic acid, l-aspartic acid and glycine. The genomic G+C content of strain H23T48T is 59.5 mol% and the average nucleotide identity value between H23T48T and F. salsibiostraticola KCT C33148T (=EBR4-1-2T) is 75.5 %. Based on the obtained data, strain H23T48T represents a novel species of the genus Flaviflexus, for which the name Flaviflexus ciconiae sp. nov. is proposed. The type strain is H23T48T (=KCTC 49253T=JCM 33282T).

New Determinants of Aminoglycoside Resistance and Their Association with the Class 1 Integron Gene Cassettes in Trueperella pyogenes.

Trueperella pyogenes is an important opportunistic animal pathogen. Different antimicrobials, including aminoglycosides, are used to treat T. pyogenes infections. The aim of the present study was to evaluate aminoglycoside susceptibility and to detect aminoglycoside resistance determinants in 86 T. pyogenes isolates of different origin. Minimum inhibitory concentration of gentamicin, streptomycin, and kanamycin was determined using a standard broth microdilution method. Genetic elements associated with aminoglycoside resistance were investigated by PCR and DNA sequencing. All studied isolates were susceptible to gentamicin, but 32.6% and 11.6% of them were classified as resistant to streptomycin and kanamycin, respectively. A total of 30 (34.9%) isolates contained class 1 integrons. Class 1 integron gene cassettes carrying aminoglycoside resistance genes, aadA11 and aadA9, were found in seven and two isolates, respectively. Additionally, the aadA9 gene found in six isolates was not associated with mobile genetic elements. Moreover, other, not carried by gene cassettes, aminoglycoside resistance genes, strA-strB and aph(3')-IIIa, were also detected. Most importantly, this is the first description of all reported genes in T. pyogenes. Nevertheless, the relevance of the resistance phenotype to genotype was not perfectly matched in 14 isolates. Therefore, further investigations are needed to fully explain aminoglycoside resistance mechanisms in T. pyogenes.

Bladder bacterial diversity differs in continent and incontinent women: a cross-sectional study.

BACKGROUND: Since the discovery of the bladder microbiome (urobiome), interest has grown in learning whether urobiome characteristics have a role in clinical phenotyping and provide opportunities for novel therapeutic approaches for women with common forms of urinary incontinence. OBJECTIVE: This study aimed to test the hypothesis that the bladder urobiome differs among women in the control cohort and women affected by urinary incontinence by assessing associations between urinary incontinence status and the cultured urobiome. STUDY DESIGN: With institutional review board oversight, urine specimens from 309 adult women were collected through transurethral catheterization. These women were categorized into 3 cohorts (continent control, stress urinary incontinence [SUI], and urgency urinary incontinence [UUI]) based on their responses to the validated Pelvic Floor Distress Inventory (PFDI) questionnaire. Among 309 women, 150 were in the continent control cohort, 50 were in the SUI cohort, and 109 were in the UUI cohort. Symptom severity was assessed by subscale scoring with the Urinary Distress Inventory (UDI), subscale of the Pelvic Floor Distress Inventory. Microbes were assessed by expanded quantitative urine culture protocol, which detects the most common bladder microbes (bacteria and yeast). Microbes were identified to the species level by matrix-assisted laser desorption and ionization time-of-flight mass spectrometry. Alpha diversity indices were calculated for culture-positive samples and compared across the 3 cohorts. The correlations of UDI scores, alpha diversity indices, and species abundance were estimated. RESULTS: Participants had a mean age of 53 years (range 22-90); most were whites (65%). Women with urinary incontinence were slightly older (control, 47; SUI, 54; UUI, 61). By design, UDI symptom scores differed (control, 8.43 [10.1]; SUI, 97.95 [55.36]; UUI, 93.71 [49.12]; P<.001). Among 309 participants, 216 (70%) had expanded quantitative urine culture-detected bacteria; furthermore, the urinary incontinence cohorts had a higher detection frequency than the control cohort (control, 57%; SUI, 86%; UUI, 81%; P<.001). In addition, the most frequently detected species among the cohorts were as follows: continent control, Lactobacillus iners (12.7%), Streptococcus anginosus (12.7%), L crispatus (10.7%), and L gasseri (10%); SUI, S anginosus (26%), L iners (18%), Staphylococcus epidermidis (18%), and L jensenii (16%); and UUI, S anginosus (30.3%), L gasseri (22%), Aerococcus urinae (18.3%), and Gardnerella vaginalis (17.4%). However, only Actinotignum schaalii (formerly Actinobaculum schaalii), A urinae, A sanguinicola, and Corynebacterium lipophile group were found at significantly higher mean abundances in 1 of the urinary incontinence cohorts when compared with the control cohort (Wilcoxon rank sum test; P<.02), and no individual genus differed significantly between the 2 urinary incontinence cohorts. Both urinary incontinence cohorts had increased alpha diversity similar to continent control cohort with indices of species richness, but not evenness, strongly associated with urinary incontinence. CONCLUSION: In adult women, the composition of the culturable bladder urobiome is associated with urinary incontinence, regardless of common incontinence subtype. Detection of more unique living microbes was associated with worsening incontinence symptom severity. Culturable species richness was significantly greater in the urinary incontinence cohorts than in the continent control cohort.

Mycetoma in Uganda: A neglected tropical disease.

Mycetoma is considered a neglected tropical disease globally. However, data on its burden and the associated complications in Uganda are limited. Hence we aimed to estimate its burden in Uganda. Firstly, a systematic PubMed search for all studies of any design on mycetoma in Uganda without restriction to the year of publication was conducted. A retrospective review of all the biopsy reports at the Pathology Reference Laboratory, Department of Pathology, Makerere University, Kampala, Uganda from January 1950 to September 2019 was conducted to identify any reports on mycetoma histological diagnosis. During the 70-years study period, 30 cases were identified by the literature review, with 249 additional cases identified by review of biopsy reports (total of 279 cases). The average incidence was estimated at 0.32/100,000 persons and prevalence of 8.32/100,000 persons per decade. However, there was a general decline in the number of cases detected recently. Males and the age group of 21-30 years were the most affected by mycetoma in Uganda, and only 7% of the cases were children. The highest number of cases was recorded from Kampala (n = 30) and Jinja (n = 19) districts. The majority of the cases (68%) were referred from surgical units. The foot was the most affected part of the body (72%). Ten per cent of the cases had bone involvement of which 58% required amputation. Fungi were the most common causative agents (89%) followed by Nocardia species (5%) and Actinomycetes (4%). The index of clinical suspicion of mycetoma was low (45%) with a very large differential diagnosis. Mycetoma is a relatively rare disease in Uganda, mostly caused by fungi, and there is a big gap in data and epidemiological studies. More systematic studies are warranted to define the true burden of mycetoma in Uganda.

Antimicrobial susceptibility of Trueperella pyogenes isolated from food-producing ruminants.

A total of 96 Trueperella pyogenes isolates, an opportunistic pathogen of food-producing ruminants, obtained from cattle (n = 34), sheep (n = 35) and goats (n = 27), and identified by Real Time PCR (qPCR), were analysed to determine the susceptibility to 12 antimicrobials commonly used in livestock, using a broth microdilution. The Minimal Inhibitory Concentration (MIC) distribution was unimodal for half of the antimicrobials tested with the exception of apramycin, gentamicin, streptomycin, oxytetracycline, tylosin, and erythromycin all of which showed bimodal MIC distributions. Low MIC90 values for penicillin, amoxicillin, ceftiofur, enrofloxacin, and gentamicin (<1 µg/ml) were obtained, suggesting that these antimicrobials would be the most effective first line empiric treatment for T. pyogenes infections in livestock. Furthermore, according to the specific T. pyogenes breakpoints for penicillin, sulfamethoxazole/trimethoprim and erythromycin, 93.7 % of isolates were susceptible to penicillin and 77.2 % to erythromycin, whereas 92.7 % were non-susceptible to sulfamethoxazole/trimethoprim. Significant differences were observed in the MIC distribution of almost all antimicrobials, except enrofloxacin, tylosin and erythromycin against cattle, sheep or goat isolates, although all antimicrobials showed similar MIC90 values, except apramycin and oxytetracycline that showed higher values when tested against cattle isolates. These data provide interesting information on the antimicrobials of choice for the treatment of infections caused by T. pyogenes in ruminants.