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In the present study, a single Arcanobacterium (A.) pinnipediorum strain isolated from discharge of a jaw swelling of a grey seal pup (Halichoerus grypus) in England, UK, was identified. This strain was further characterized by phenotypical investigations, by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS), by Fourier transform infrared spectroscopy (FT-IR), and genotypically by sequencing the 16S rRNA gene and the genes gap encoding glyceraldehyde 3-phosphate dehydrogenase, tuf encoding elongation factor tu, and rpoB encoding the β subunit of bacterial RNA polymerase. The present study gives a first detailed characterization of the species A. pinnipediorum from a grey seal in the UK. However, the route of infection of the grey seal with the bacterial pathogen remains unclear.

Trueperella (T.) bernardiae is a well-known bacterial pathogen in infections of humans, rarely in animals. In the present study, five T. bernardiae isolates, isolated from five Peking ducks of four different farms, were identified by phenotypic properties, by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) analysis, and genotypically by sequencing the 16S ribosomal RNA (rRNA) gene, the superoxide dismutase A encoding gene sodA, and the glyceraldehyde-3-phosphate dehydrogenase encoding gene gap. In addition, the T. bernardiae isolates could be identified with a newly developed loop-mediated isothermal amplification (LAMP) assay based on the gyrase encoding housekeeping gene gyrA. All these tests clearly identified the T. bernardiae isolates to the species level. However, the detection of the specific gene gyrA with the newly designed LAMP assay appeared with a high sensitivity and specificity, and could help to identify this bacterial species in human and animal infections in future. The importance of the T. bernardiae isolates for the clinical condition of the ducks and for the problems at farm level remains unclear.

Submitted 2020-06-30 | Accepted 2020-07-25 | Available 2020-12-01 https://doi.org/10.15414/afz.2020.23.mi-fpap.67-73 Milk is considered not only a source of nutrient for the offspring but also a font of immunoregulatory compounds capable to predispose the naive intestinal immune system of the new-born to react to the external environment. In the present study we evaluated the composition of milk microbiome from cows classified according to milk total and differential somatic cell counts. A total of 34, 13 and 13 milk samples of healthy, at risk and subclinical or chronic cows, respectively, were collected during the same milking from a local dairy herd. Through Next Generation Sequencing (NGS) of bacterial 16S rRNA gene, the differences of taxa in terms of relative abundances (RA) and alpha and beta biodiversity were analysed. The RA of several genera were statistically significant in the three groups, such as Arcanobacterium (p=0.001), Rhodococccus (p

Summary Introduction Aging and comorbidities such as diabetes and vascular problems contribute to the increasing occurrence of chronic wounds. From the beginning of 2016, a marked increase in Arcanobacterium haemolyticum (ARH) in chronic wound cultures was noted among patients visiting a wound expertise centre in The Netherlands. Aim To report the outbreak investigation of ARH cultured from chronic wounds and describe the implemented infection prevention measures. Methods In total, 50 ARH isolates were sent to a reference laboratory for molecular typing. Samples for bacterial culture and ARH polymerase chain reaction were taken from care workers, the environment and items used for wound care. Infection prevention measures were implemented in a bundled approach, involving education, better aseptic wound care conditions and hygienic precautions. Before and after the implementation of infection prevention measures, two screening rounds of ARH testing were performed among all patients receiving home care. Results ARH isolates from wound care patients were found to be identical by core genome multi-locus sequence typing. No definite outbreak source could be determined by culture. However, three pairs of forceps, used by two nurses on multiple patients, were found to be ARH positive by polymerase chain reaction. In the two screening rounds before and after the implementation of infection prevention measures, the proportion of ARH-positive patients decreased significantly from 20% (20/99) to 3% (3/104). Subsequently, no new cases occurred. Conclusion This first ARH outbreak was likely caused by re-using contaminated instruments. Through the implementation of improved infection prevention measures and re-education of all employees involved, the outbreak was controlled. With the current trend of care transition, infection control must be a major concern.

A polyphasic taxonomic study was performed on an unidentified Arcanobacterium -like Gram-stain-positive bacterium designated strain C605018/01/1T isolated from a milk sample collected from the udder of a cow at post mortem. Comparative 16S rRNA gene sequencing showed that the bacterium belonged to the genus Arcanobacterium and was most closely related to the type strain of Arcanobacterium pluranimalium (99.76 %); sequence similarities to all other Arcanobacterium species were below 97 %. The wet-lab DNA–DNA hybridization values among strain C605018/01/1T and A. pluranimalium DSM 13483ᵀ were low, 16.9 % (reciprocal, 49.8 %). Pertaining to the whole genome sequence with a total length of 2.02 Mb and 1654 protein counts, the novel strain C605018/01/01T displayed a G+C content of 51.6 % mol%. The presence of the major menaquinone MK-9(H4) supported the affiliation of this strain to the genus Arcanobacterium . The polar lipid profile consisted of the major components diphosphatidylglycerol, phosphatidylcholine, phosphatidylinositol, phosphatidylinositol-mannoside and unidentified glycolipid and aminophospholipids. Based on these results it is proposed that strain C605018/01/1T should be classified as representing a novel species, Arcanbacterium bovis sp. nov. The type strain C605018/01/1T (CCUG 45425T=DSM 107286T=BCCM/LMG 30783T)

The first description of Trueperella (T.) abortisuis was presented in Japan in 2009 by Azuma and colleagues. In the current study, eight T. abortisuis strains were identified by a newly developed loop-mediated isothermal amplification (LAMP) assay based on the glyceraldehyde 3-phosphate dehydrogenase (GAPDH) encoding gene gap. Two T. abortisuis strains were obtained from prepuce of a seven-month-old boar and pooled foetal stomach contents in the United Kingdom, while the other six T. abortisuis strains were recovered from aborted foetal material of six pigs from a single farm in Germany. The developed LAMP assay showed an analytical sensitivity of 22 pg μL

The present study reports the isolation of A. hippocoleae from genital swabs of 15 apparently healthy mares (at least one had an abortion one month earlier) and describes the genotypic and phenotypic characterisation of these strains. The mares were of eight different breeds with a thoroughbred dominance and came from 11 breeding farms located in the French region of Brittany. 16S rRNA gene sequencing confirmed the species' identification by comparing it with reference strain A. hippocoleae CIP 106850T. Some degree of natural divergence within A. hippocoleae was observed by 16S rRNA sequencing (two 1,002-pb sequences), MALDI-TOF MS typing (two groups), a CAMP test (three different intensities of haemolysis from CAMP-positive results) and API® Coryne system (five profiles). The strains were all susceptible to the antimicrobials tested. A national prevalence survey would be required to estimate the frequency of A. hippocoleae carriage in mares and stallions and to verify the presence of A. hippocoleae outside the French region of Brittany, which is the only one found to be affected in the current study, probably because the isolates were recovered from a single field laboratory in this region.

Background: Emerging evidence has emphasized the functional of microbiota in the etiopathogenesis of rheumatoid arthritis (RA). Previously we and other groups have demonstrated that the urinary infection was implicated in the pathogenesis of RA. However, the profile of urinary microbiota and its association with RA remains to be investigated. Methods: The study cohort consisted of 39 patients with RA, including 8 treatment-naive patients, and 37 age- and sex-matched healthy controls. The clean midstreams of urine samples were collected from both groups. The microbial profiles and bacterial loads of urinary microbiota were identified and quantified by 16S ribosomal RNA (16S rRNA) gene sequencing and quantitative PCR, respectively. The compositions of urinary microbiota and its associations with clinical and molecular characteristics were analyzed. The urinary metabolites were detected by gas chromatography. Findings: Compared to healthy controls, the urinary microbiota from RA patients displayed an increase in microbial richness with a decrease in microbial dissimilarity. There were 40 altered genera with absolute quantities in patients with RA, including 31 enriched (Proteus, Faecalibacterium, Bacteroides, etc.) and 9 deficient genera (Arcanobacterium, Rothia, Megasphaera, Ureaplasma, etc.). These RA-associated urinary taxa achieved a performance power of 84.6% for discriminating RA patients from healthy controls. Notably, the RA-enriched, but not deficient genera, were correlated with elevated disease duration, anti-CCP antibodies, joint damages, and increase of circulating LPS binding protein (LBP) and soluble CD14. Furthermore, the compositions of the urinary metabolites, including proline, citric acid and oxalic acid, were significantly different in RA patients from healthy controls. These RA-associated urinary metabolites showed close correlations with urinary microbiota. Interpretation: An increased microbial richness and shifted compositional profiles are found in the urinary microbiota of RA patients, which is strongly associated with immunological and metabolic changes in RA. Funding: National Natural Science Foundation of China and Beijing Municipal Science & Technology Commission. Declaration of Interest: None to declare. Ethical Approval: Approval for the study was obtained from the Medical Ethics Committee of Peking University People’s Hospital.

Simple Summary Knowing which bacteria dominate vaginal microbiota and its variation throughout the cycle is important to study how to prevent reproductive diseases. In women, vaginal microbiota is dominated by Lactobacillus but this does not happen in other animals. Little is known about equine vaginal microbiota. The aim of this study was to describe the dynamics of equine vaginal microbiota during the ovarian cycle. Eight healthy adult Arabian mares were used to characterize vaginal microbiota by standard microbiologic and metagenomic procedures. The abundance of Lactobacillus was < 2% by both methods, meaning that equine vaginal microbiota was not dominated by these bacteria. Dominant bacteria included other genera such as Porphyromonas and Campylobacter among others. No changes in vaginal microbiota composition were found, suggesting that equine vaginal microbiota was stable throughout the ovarian cycle. Abstract Lactic acid bacteria (LAB) dominate human vaginal microbiota and inhibit pathogen proliferation. In other mammals, LAB do not dominate vaginal microbiota, however shifts of dominant microorganisms occur during ovarian cycle. The study objectives were to characterize equine vaginal microbiota in mares by culture-dependent and independent methods and to describe its variation in estrus and diestrus. Vaginal swabs from 8 healthy adult Arabian mares were obtained in estrus and diestrus. For culture-dependent processing, bacteria were isolated on Columbia blood agar (BA) and Man Rogosa Sharpe (MRS) agar. LAB comprised only 2% of total bacterial isolates and were not related to ovarian phases. For culture-independent processing, V3/V4 variable regions of the 16S ribosomal RNA gene were amplified and sequenced using Illumina Miseq. The diversity and composition of the vaginal microbiota did not change during the estrous cycle. Core equine vaginal microbiome consisted of Firmicutes, Bacteroidetes, Proteobacteria and Actinobacteria at the phylum level. At the genus level it was defined by Porphyromonas, Campylobacter, Arcanobacterium, Corynebacterium, Streptococcus, Fusobacterium, uncultured Kiritimatiaellae and Akkermansia. Lactobacillus comprised only 0.18% of the taxonomic composition in estrus and 0.37% in diestrus. No differences in the relative abundance of the most abundant phylum or genera were observed between estrus and diestrus samples.

We report the case of a 90-year-old female who was admitted to our hospital due to a three-day history of right abdominal pain and fever of 39 °C (102 °F). The patient's blood pressure was low, with good blood perfusion and no jaundice, and her abdomen was soft and tender in the right hypochondriac and lateral region, with no guarding. Laboratory tests showed: blood glucose level of 201 mg/dl, 362 U/l AST, 237 U/l ALT, 2.5 mg/dl bilirubin, 237 U/l alkaline phosphatase and leukocytosis associated with a left shift. An abdominal ultrasound scan showed a collection of echogenic material and a shadow suggestive of air in hepatic segment 3. Meropenem and metronidazole treatment was started after taking blood cultures, which were negative. A computed tomography (CT) scan confirmed the presence of a liver abscess in segment 3, containing a high-density linear image.

We report the case of a twenty-year-old immunocompetent male patient presentingto the emergency room with pharyngitis and fever. Blood cultures were drawn and Arcanobac-terium haemolyticum (rough biotype) was recovered. The presence of the arcanolysin gene wasinvestigated at the molecular level and the upstream region was amplified and sequenced inorder to correlate it with the smooth or rough biotype. Although the isolate was susceptible topenicillin, vancomycin and gentamicin, empirical treatments first with amoxicillin/clavulanicacid (1 g/12 h) and then with ceftriaxone (1 g/12 h) failed and the infection evolved to sep-sis. Finally, treatment with vancomycin (1 g/12 h) plus piperacillin/tazobactam (4.5 g/8 h) waseffective. Lemierre?s syndrome was ruled out. To the best of our knowledge, this is the firstcase of bacteremia by A. haemolyticum reported in Argentina. Fil: Verona, Julián. Universidad de Buenos Aires; Argentina Fil: Dirialdi, Noelia Cecilia. Universidad de Buenos Aires; Argentina Fil: Mutti, María Virginia. Universidad de Buenos Aires; Argentina Fil: Scioli, Marcelo Carlos. Universidad de Buenos Aires; Argentina Fil: Thougnon Islas, Fernando Andrés. Universidad de Buenos Aires; Argentina Fil: Curi Antún, Germán Emilio. Universidad de Buenos Aires; Argentina Fil: Sgariglia, Lisandro Juan. Universidad de Buenos Aires; Argentina Fil: Homar, María Guillermina. Universidad de Buenos Aires; Argentina Fil: Legato, José Andrés. Universidad de Buenos Aires; Argentina Fil: Costa, Agustina. Universidad de Buenos Aires; Argentina Fil: Maldonado, María Laura. Gobierno de la Ciudad de Buenos Aires. Hospital de Pediatría "Juan P. Garrahan"; Argentina. Universidad de Buenos Aires; Argentina Fil: Di Conza, José Alejandro. Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay; Argentina Fil: Litterio Bürki, Mirta R.. Universidad de Buenos Aires; Argentina

The use of smokeless tobacco products (STPs) can cause many serious health problems. The oral microbiota plays important roles in oral and systemic health, and the disruption in the oral microbial population is linked to periodontal disease and other health problems. To assess the impact of smokeless tobacco on oral microbiota in vivo, high-throughput sequencing was used to examine the oral microbiota present in Syrian Golden hamster cheek pouches. Sixteen hamsters were divided into four groups and treated with the STP Grizzly snuff (0, 2.5, 25, or 250 mg) twice daily for 4 weeks. After 0, 1, 2, 3, and 4 weeks of treatment, bacterial genomic DNA was extracted from oral swabs sampled from the cheek pouches of the hamsters. The oral bacterial communities present in different hamster groups were characterized by sequencing the hypervariable regions V1-V2 and V4 of 16S rRNA using the Illumina MiSeq platform. Fifteen phyla, 27 classes, 59 orders, 123 families, and 250 genera were identified from 4,962,673 sequence reads from the cheek pouch samples. The bacterial diversity and taxonomic abundances for the different treatment groups were compared to the non–treated hamsters. Bacterial diversity was significantly decreased after 4 weeks of exposure to 2.5 mg, and significantly increased by exposure to 250 mg STP. Treatment with 250 mg STP significantly increased Firmicutes, transiently increased Cyanobacteria and TM7, and decreased Bacteroidetes and Fusobacteria compared to the control group. At the genus level, 4 weeks of administration of 250 mg STP significantly increased Granulicatella, Streptococcus, Oribacterium, Anaerococcus, Acidaminococcus, Actinomyces, Eubacterium, Negativicoccus, and Staphylococcus, and decreased Bacteroides, Buleidia, Dialister, and Leptotrichia, and transiently decreased Arcanobacterium compared to the control group. For the first time, an animal model was used for evaluating the effects of STP on oral microbiota by metagenomic sequencing. Our results provide a view of the shift of the oral microbiota in response to STP exposure in Syrian Golden hamster. Our findings indicate that the use of smokeless tobacco significantly disrupts the oral microbiota.

An 81-year-old Japanese man with no history of diabetes mellitus was admitted to our hospital for a fever with a new ulcerative lesion on the left heel. Blood cultures on admission grew Arcanobacterium haemolyticum in aerobic bottles. He was therefore diagnosed with A. haemolyticum bacteremia and osteomyelitis complicated with foot decubitus ulcer. He was successfully treated with intravenous antibiotic therapy and debridement of the left heel. Our case and literature review show that it is important to recognize that A. haemolyticum is a systemic causative pathogen in immunocompetent patients in primary care practice.

Introduction Arcanobacterium haemolyticum causes pharyngotonsillitis in children and young adults. It is rarely isolated in pharyngeal swabs as testing for it is not routine. Data on complications, management, and antibiotic susceptibility testing is limited. We sought to review the available literature on the presentation and management of A. haemolyticum pharyngotonsillitis in this age group. Methods A systematic review of eligible studies reporting pharyngotonsillitis and related complications in children and young adults caused by A. haemolyticum was conducted. Literature from case reports, case series, and available cohorts was compiled. Data were analyzed using descriptive statistics. Results The initial database search yielded 63 articles, after applying exclusion criteria 17 studies were included. 191 patients were identified with a median age of 16.5 years. The most common presentation was throat pain reported in 93.7% of patients. Tonsillar exudates, fever at presentation and rash were present in more than half of the reviewed cases. The diagnosis was established by a positive culture on a pharyngeal swab in 98.8% of swabs collected. Complications described included peritonsillar abscesses, Lemierre's syndrome, pneumonia, and sepsis. Penicillin was the first line antibiotic in 81% of patients followed by macrolides in 19 patients (18%). Conclusions A. haemolyticum occurs in children and young adults and may result in complications. Our review supports its susceptibility to penicillin. We suggest a diagnostic and management algorithm to guide clinicians in targeted testing and aid with decision making regarding timely and appropriate antibiotic therapy, in an effort to reduce the burden of its complications.

Background Infectious skin disorders are not uncommon in mink. Such disorders are important as they have a negative impact on animal health and welfare as well as on the quality and value of the fur. This study presents the isolation of Arcanobacterium phocae from mink with severe skin lesions and other pathological conditions, and from wild seals and otters. Results In 2015, A. phocae was isolated for the first time in Denmark from outbreaks of dermatitis in mink farms. The outbreaks affected at least 12 farms. Originating from these 12 farms, 23 animals cultured positive for A. phocae. The main clinical findings were necrotizing pododermatitis or dermatitis located to other body sites, such as the lumbar and cervical regions. A. phocae could be isolated from skin lesions and in nine animals also from liver, spleen and lung, indicating a systemic spread. The bacterium was also, for the first time in Denmark, detected in dead seals (n = 9) (lungs, throat or wounds) and otters (n = 2) (throat and foot). Conclusions An infectious skin disorder in mink associated with A. phocae has started to occur in Danish farmed mink. The origin of the infection has not been identified and it is still not clear what the pathogenesis or the port of entry for A. phocae infections are. Electronic supplementary material The online version of this article (doi:10.1186/s13028-017-0342-8) contains supplementary material, which is available to authorized users.

Trueperella pyogenes is an opportunistic pathogen that causes diverse pyogenic infections in livestock. The genes that encode the exotoxin pyolysin (plo) and other putative factors that promote adhesion of pathogen to host cells (fimbriae fimA, fimC, fimE, fimG, neuraminidases nanH, nanP, and collagen-binding protein cbpA) have been associated with virulence, particularly in mastitis and uterus infections of dairy cows. However, the role of these virulence markers in the pathogenicity of the agent in domestic animals infections still is incompletely understood. The genes plo, fimA, fimC, fimE, fimG, nanH, nanP, and cbpA were investigated in 71 T. pyogenes strains recovered from cattle, sheep, goats, dogs, equines, and a pig, recovered from mastitis (n = 35), and non-mastitis (n = 36) cases (abscesses, reproductive tract diseases, pneumonia, lymphadenitis, encephalitis). The most common genes harboured by the isolates were: plo (71/71 = 100·0%), fimA (70/71 = 98·6%), nanP (56/71 = 78·9%), fimE (53/71 = 74·6%), fimC (46/71 = 64·8%) and nanH (45/71 = 63·4%), whereas cbpA (6/71 = 8·4%) and fimG (4/71 = 5·6%) were uncommon. The most frequent genotypes were plo/fimA/fimE/fimC/nanH/nanP (17/71 = 23·9%), plo/fimA/fimE/nanH/nanP (13/71 = 18·3%), and plo/fimA/fimE/fimC/nanP (11/71 = 15·5%). No association was observed between the presence of genes vs clinical signs or host species. To the best of our knowledge, this is the first report on aforementioned virulence factors of pathogen detected in diseased horses and dogs. Significance and impact of the study The role of particular virulence factors of Trueperella pyogenes that determine different pyogenic infections among domestic animals is poorly understood. Eight putative virulence genes and genotype profiles of 71 isolates were investigated among different clinical manifestations in domestic animals. The most common genes were plo (71/71 = 100·0%), fimA (70/71 = 98·6%), nanP (56/71 = 78·9%), fimE (53/71 = 74·6%), fimC (46/71 = 64·8%) and nanH (45/71 = 63·4%), whereas plo/fimA/fimE/fimC/nanH/nanP (17/71 = 23·9%), plo/fimA/fimE/nanH/nanP (13/71 = 18·3%), and plo/fimA/fimE/fimC/nanP (11/71 = 15·5%) were the most frequent genotypes. Studies involving virulence factors are critical in the investigation of molecular epidemiology, pathogenicity, and hypothetical differences in the virulence among T. pyogenes strains from different geographical areas.

A taxonomic study using a polyphasic approach was performed on an unidentified Arcanobacterium -like Gram-stain-positive bacterium isolated from the genital tract of a rhinoceros. Comparative 16S rRNA gene sequencing showed that the bacterium belonged to the genus Arcanobacterium and was most closely related to the type strains of Arcanobacterium canis (98.8 % 16S rRNA gene sequence similarity), Arcanobacterium phocisimile (97.8 %), Arcanobacterium phocae (97.7 %), Arcanobacterium haemolyticum (97.4 %), Arcanobacterium hippocoleae (96.6 %), Arcanobacterium pinnipediorum (96.4 %) and Arcarnobacterium pluranimalium (95.4 %). DNA–DNA hybridization values between strain 647T and Arcanobacterium canis DSM 25104T were very low, 13.4 % (reciprocal 15.9 %). The genomic DNA G+C content of strain 647T was 58.7 mol%. The presence of the major menaquinone MK-9(H4) supported the affiliation of this strain to the genus Arcanobacterium . The polar lipid profile consisted of the major components diphosphatidylglycerol, phosphatidylcholine and an unidentified phosphoglycolipid. The results of physiological and biochemical testing clearly distinguished the unknown bacterium from other species of the genus Arcanobacterium . Based on these tests, it is proposed that the unknown bacterium should be classified as a representative of a novel species of the genus Arcanobacterium named Arcanobacterium wilhelmae sp. nov. The type strain is 647T (=DSM 102162T=LMG 29418T).

Sphingomyelinases D have only been identified in arachnid venoms, Corynebacteria, Arcanobacterium, Photobacterium and in the fungi Aspergillus and Coccidioides. The arachnid and bacterial enzymes share very low sequence identity and do not contain the HKD sequence motif characteristic of the phospholipase D superfamily, however, molecular modeling and circular dichroism of SMases D from Loxosceles intermedia and Corynebacterium pseudotuberculosis indicate similar folds. The phospholipase, hemolytic and necrotic activities and mice vessel permeabilities were compared and both enzymes possess the ability to hydrolyze phospholipids and also promote similar pathological reactions in the host suggesting the existence of a common underlying mechanism in tissue disruption. J. Cell. Biochem. 118:2053-2063, 2017. © 2016 Wiley Periodicals, Inc.

Dry-cured hams, shoulders and loins of Iberian pigs are highly appreciated in national and international markets. Salting, additive addition and dehydration are the main strategies to produce these ready-to-eat products. Although the dry curing process is known to reduce the load of well-known food borne pathogens, studies evaluating the viability of other microorganisms in contaminated pork have not been performed. In this work, the efficacy of the dry curing process to eliminate three swine pathogens associated with pork carcass condemnation, Streptococcus suis, Streptococcus dysgalactiae and Trueperella pyogenes, was evaluated. Results of this study highlight that the dry curing process is a suitable method to obtain safe ready-to-eat products free of these microorganisms. Although salting of dry-cured shoulders had a moderate bactericidal effect, results of this study suggest that drying and ripening were the most important stages to obtain dry-cured products free of these microorganisms.

Culturomics studies the microbial variety of the human microbiome by combining diversified culture conditions, matrix-assisted laser desorption/ionization time-of-flight mass spectrometry and 16S rRNA gene identification. This study identifies three putative new bacterial species: Arcanobacterium ihumii sp. nov. strain Marseille-P5647T, Varibaculum vaginae sp. nov. strain Marseille-P5644T and Tessaracoccus timonensis sp. nov. strain Marseille-P5995T, which we describe according to the concept of taxonogenomics. Keywords: Arcanobacterium ihumii sp. nov., culturomics, Tessaracoccus timonensis sp. nov., vagina, Varibaculum vaginae sp. nov.

A Gram‐staining positive facultative anaerobic, non‐motile strain, sk1b4T, was isolated from canker of symptomatic bark tissue of a Populus × euramericana. 16S rRNA gene sequence analyses showed that strain sk1b4T shared the highest similarity with Arcanobacterium phocisimile (94.1%). Within the phylogenetic tree, the novel isolate formed a distinct branch from Actinobaculum, Arcanobacterium, and Trueperella. The percentage of conserved proteins calculated from genomic sequence indicated a low level of relatedness between the novel strain and its phylogenetic neighbors. Growth of the novel strain occurred at temperatures between 10 and 41°C, and within a pH range of 6.0–9.0; optimal growth occurred at 30°C and at pH 6.0–9.0. Growth also occurred within a NaCl concentration of 1%–5% (w/v). The major fatty acids of the strain were C14:0, C16:0, and C18:1 ω9c , and major polar lipids were glycolipid, phosphatidylinositol mannoside, phospholipid, diphosphatidylglycerol, and phosphatidylglycerol. Respiratory quinone was absent. On the basis of phenotypic and genotypic characteristics, we propose that the novel isolate should be classified as a novel species in a new genus: Ancrocorticia populi gen. nov., sp. nov. The type strain is sk1b4T (=CFCC 14564T= KCTC 39919T).

We investigated the infection of Bovine Leukemia Virus (BLV) related to other pathogens [Neospora caninum, Bovine Herpesvirus-1 (BoHV-1), Bovine Viral Diarrhea Virus (BVDV), and pathogenic bacteria] in 80 bovine aborted fetuses. The materials comprised whole fetuses, fetal organs, and placenta. The BLV was diagnosed by nested-PCR (env gp51 BLV gene), the identification of viral genotypes by sequencing, and the phylogenetic analysis by neighborjoining and maximum composite likelihood methods. The other pathogens and diagnoses were, respectively: Neospora caninum (nested-PCR), BoHV-1 (nested-PCR), BVDV (PCR), Brucella spp. (isolation and identification), Leptospira spp. (PCR), aerobic bacteria [Enterobacteriaceae, Gram positive cocci, Trueperella (Arcanobacterium) pyogenes] and micro-aerophilic (Campylobacter spp., Histophilus somni, and Listeria monocytogenes) by isolation and identification. BLV fetal antibodies were identified by ELISA kit. Thirteen (16.25%) fetuses were positive by BLV nested-PCR. Phylogenetic analysis revealed BLV genotypes 1, 5, and 6, which are frequently found in cattle in Brazil, Argentina, and Uruguay. No fetuses were positive for BLV antibodies by ELISA. A single case of coinfection with BLV was found for each of the pathogens Trueperella (Arcanobacterium) pyogenes, Klebsiella spp., and Streptococcus spp. were isolated as a pure or representing the preponderance of bacteria in a pooled culture. In the 67 BLV-negative fetuses, pathogens identified were single cases of Trueperella (Arcanobacterium) pyogenes, Staphylococcus aureus, and Brucella abortus; 2 of Escherichia coli; 3 of bovine viral diarrhea virus; and 4 of Neospora caninum. No pathogens were found in 55 fetuses. The low number of BLV positive samples infected or no by other pathogens didn´t allow performing statistical analysis for understanding if there were significative differences among not infected and infected BLV fetuses. Because BLV is an immunosuppressive agent and predisposes the cow to other pathogens, its connection with Leukemia or abortions need additional studies with bigger sampling, for elucidating pathogenesis in the pregnant cow and in the fetus. The rates of BLV transplacental transmission show the necessity of prophylactic measures in Brazilian cattle herds, in order to avoid infection in utero.

We investigated the effects of early intervention with maternal fecal microbiota and antibiotics on gut microbiota and the metabolites. Five litters of healthy neonatal piglets (Duroc &times, Landrace &times, Yorkshire, nine piglets in each litter) were used. Piglets in each litter were orally treated with saline (CO), amoxicillin treatment (AM), or maternal fecal microbiota transplantation (MFMT) on days 1&ndash, 6, with three piglets in each treatment. Results were compared to the CO group. MFMT decreased the relative abundances of Clostridium sensu stricto and Parabacteroides in the colon on day 7, whereas the abundance of Blautia increased, and the abundance of Corynebacterium in the stomach reduced on day 21. AM reduced the abundance of Arcanobacterium in the stomach on day 7 and reduced the abundances of Streptococcus and Lachnoclostridium in the ileum and colon on day 21, respectively. The metabolite profile indicated that MFMT markedly influenced carbohydrate metabolism and amino acid (AA) metabolism on day 7. On day 21, carbohydrate metabolism and AA metabolism were affected by AM. The results suggest that MFMT and AM discriminatively modulate gastrointestinal microflora and alter the colonic metabolic profiles of piglets and show different effects in the long-term. MFMT showed a location-specific influence on the gastrointestinal microbiota.

Trueperella bernardiae is a nonspore‐forming, nonmotile, facultative anaerobic, gram‐positive coccobacilli; it is catalase and oxidase negative and has variable hemolytic activity. Colonies are circular, smooth, and slightly convex with a glassy appearance and diameters range from 0.2 to 0.5 mm 1. This bacterium was first described as a likely opportunistic pathogen based on its morphological, biochemical, and chemical properties as part of Coryneform Group 2 2. This group was characterized using biochemical and chemotactic methods (16S rRNA sequence analysis) from bacteria isolated from blood samples and human abscesses and the name Actinomyces bernardiae sp. nov. was proposed 3. It was classified as part of the genus Arcanobacterium 1 and was later reclassified as part of the genus Trueperella 4. There are some documented cases of infection caused by Trueperella bernardiae. These bacteria usually come from the skin as it can be part of the regular skin flora 5. Trueperella bernardiae was previously identified from blood culture, abscesses, and eye infections 3. There are case reports of severe urinary tract infections complicated by septicemia 6, septic arthritis 7, coinfection with Staphylococcus aureus from chronic osteitis 8, prosthetic joint infection 9, necrotizing fasciitis 10, bacteremia secondary to soft tissue infection 11, bacteremia due to chronic ulcers of the lower limbs 5, diabetic foot infection, and sepsis 12. As far as we know, this report describes the first case of wound infection caused by Trueperella bernardiae after laparoscopic cholecystectomy.

Background An adult male galago (Otolemur garnettii) presented for fight wounds following pairing for breeding. Treatment was symptomatic with recovery. Following resolution, the animal re-presented and died, despite additional treatment. Methods Necropsy, histopathology, bacterial cultures, and 16S RNA sequencing. Results A large intrathoracic/intra-abdominal abscess due to Trueperella pyogenes was found at necropsy. Conclusions T. pyogenes should be considered in abscesses/wounds of galagos.

The objectives of this study were to report the prevalence of Escherichia coli and Trueperella pyogenes in the uterus of postpartum dairy cows before the onset of postpartum metritis (PPM) and to quantify their association with subsequent occurrence of PPM, to quantify the association between the presence of genes encoding E. coli virulence factors (VF) and PPM, and to determine the accuracy of using early postpartum uterine bacteriology results (bacteria and VF) to identify cows at risk of PPM. A prospective cohort study was conducted on 3 commercial dairy farms. Uterine swabs were collected from 371 Holstein dairy cows (3 commercial herds) at 1 to 7d in milk and submitted to the laboratory for identification of E. coli, T. pyogenes, and E. coli VF. A total of 40 VF were tested using the radioactive probe hybridization method. Postpartum metritis was defined as the presence of a fetid watery red-brown uterine discharge, associated with fever (rectal temperature >39.5°C), and systemic signs of illness (dullness, reduced appetite, and milk production). Surveillance of PPM was done by trained farmers blinded to laboratory results and cows were followed until 21d in milk. Statistical analyses were conducted using 2×2 tables and mixed logistical regression models. Prevalences of E. coli, T. pyogenes, and PPM were 42, 34, and 15%, respectively. A total of 32 VF were found in E. coli isolates. Most prevalent VF were extraintestinal pathogenic genes such as fimH (89%), hlyE (87%), and iss (70%). Cows positive for intrauterine E. coli were 3.2 times more likely to have subsequent PPM compared with bacteriologically negative cows. Cows with VF hra1 in their uterus were 2.7 times more likely to have PPM than cows positive for E. coli and negative for hra1 and 5.9 times more likely than bacteriologically negative cows. Cows with VF kpsMTII in their uterus were 3.2 times more likely to have PPM than cows positive for E. coli and negative for kpsMTII and 6.2 times more likely than bacteriologically negative cows. Using E. coli, hra1, and kpsMTII as predictors for subsequent PPM, positive predictive values were 23, 31, and 42%, respectively, whereas the negative predictive values were 91, 80, and 78%, respectively. Overall, these results showed that E. coli and some VF were associated with PPM.

Trueperella (T.) pyogenes is an opportunistic pathogen that causes suppurative diseases in domestic animals. In this work, the properties, pathogenesis and phenotypic diversity of T. pyogenes isolates from bovine mastitis were studied. Both pyolysin (plo) and collagen-binding protein (cbp) virulence factor genes were detected by PCR in all T. pyogenes isolates (n = 50). Using the tissue culture plate method, 90% of T. pyogenes isolates were able to form biofilms. The minimum inhibitory concentrations (MICs) of 13 antimicrobials against T. pyogenes isolates were determined. High susceptibility was observed to rifampin (96%), ampicillin (94%), ciprofloxacin (94%), and penicillin (92%), while low susceptibility was found to trimethoprim-sulphamethoxazole (10%) and bacitracin (2%). The intracellular assay revealed that T. pyogenes isolates had different cytopathogenic effects on cells. The high percentage (28.6%) of T. pyogenes isolates suggests that this bacterium is an important contributor to mastitis. Moreover, the high occurrence of multidrug resistance, biofilm production, intracellular survival, and the temporal dynamics of T. pyogenes interactions are key factors for a better understanding of how immunity acts on infections with these bacteria and how they evade immune surveillance, thus highlighting the need for the prudent use of antimicrobial agents in veterinary medicine.

A polyphasic taxonomic study was performed on an unidentified Arcanobacterium-like, Gram-stain-positive bacterium, strain 2710T, isolated from a harbour seal. Comparative 16S rRNA gene sequence analysis showed that this bacterial strain belonged to the genus Arcanobacterium and was related most closely to the type strains of Arcanobacterium phocae (98.4 % similarity) and Arcanobacterium phocisimile (97.5 %). 16S rRNA gene sequence similarities to the type strains of other Arcanobacterium species were between 95.3 and 96.9 %. DNA–DNA hybridization values between strain 2710T and A. phocae DSM 10002T and A. phocisimile LMG 27073T were 4.7 % (reciprocal 56 %) and 23 % (reciprocal 7.7 %), respectively. The presence of the major menaquinone MK-9(H4) and a polar lipid profile with the major compounds diphosphatidylglycerol, phosphatidylinositol and phosphatidylinositol mannoside supported the affiliation of strain 2710T to the genus Arcanobacterium. The major fatty acids were C16:0, C18:1ω9c, C18:0 and C18:2ω6,9c/anteiso-C18:0. The peptidoglycan structure was of cross-linkage type A5α (l-Lys–l-Lys–d-Glu). Physiological and biochemical tests clearly distinguished the isolate from other members of the genus Arcanobacterium. Based on these tests, it is proposed that this unknown bacterium should be classified as a novel species of the genus Arcanobacterium, with the name Arcanobacterium pinnipediorum sp. nov. The type strain is 2710T ( = DSM 28752T = LMG 28298T).

The present study was designed to identify nine Arcanobacterium phocae strains isolated from cases of mink dermatitis of a single farm in Finland and characterize the strains for epidemiological relationships. All nine strains and previously described A. phocae used for comparative purposes were identified and further characterized phenotypically, by matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS), by Fourier Transform Infrared Spectroscopy (FT-IR) and genotypically by detection of phocaelysin encoding gene phl with a previously developed loop-mediated isothermal amplification (LAMP) assay and by sequencing 16S rRNA gene and gene phl, the elongation factor tu encoding gene tuf and the β subunit of bacterial RNA polymerase encoding gene rpoB. Genetic relatedness among isolates was determined using whole-genome single nucleotide polymorphism (wgSNP) analysis. The wgSNP results, partly the MALDI-TOF MS and FT-IR analyses and sequencing of the genes, revealed that the nine A. phocae strains recovered from a single farm showed close sequence similarities among each other and differed from previously investigated A. phocae strains isolated from other farms and animals in Finland and from the A. phocae type strain. This indicated a close epidemiological relationship of the A. phocae strains isolated from a single farm and that the nine A. phocae strains of the present study might have developed from a common ancestor.

Systemic Arcanobacterium pyogenes is a rare bacterial infection in humans.1The diagnosis of thrombotic thrombocytopenic purpura (TTP)-like syndrome and infective endocarditis (IE) is often elusive. We report a case of TTP-like syndrome associated with A. pyogenes endocarditis in a post-allogenic transplant patient.

Preventing postpartum uterine disease depends on the ability of endometrial cells to tolerate the presence of the bacteria that invade the uterus after parturition. Postpartum uterine disease and endometrial pathology in cattle are most associated with the pathogen Trueperella pyogenes. Trueperella pyogenes secretes a cholesterol-dependent cytolysin, pyolysin, which causes cytolysis by forming pores in the plasma membrane of endometrial stromal cells. The aim of the present study was to identify cell-intrinsic pathways that increase bovine endometrial stromal cell tolerance to pyolysin. Pyolysin caused dose-dependent cytolysis of bovine endometrial stromal cells and leakage of lactate dehydrogenase into supernatants. Cell tolerance to pyolysin was increased by inhibitors that target the mevalonate and cholesterol synthesis pathway, but not the mitogen-activated protein kinase, cell cycle, or metabolic pathways. Cellular cholesterol was reduced and cell tolerance to pyolysin was increased by supplying the mevalonate-derived isoprenoid farnesyl pyrophosphate, or by inhibiting farnesyl-diphosphate farnesyltransferase 1 or geranylgeranyl diphosphate synthase 1 to increase the abundance of farnesyl pyrophosphate. Supplying the mevalonate-derived isoprenoid geranylgeranyl pyrophosphate also increased cell tolerance to pyolysin, but independent of changes in cellular cholesterol. However, geranylgeranyl pyrophosphate inhibits nuclear receptor subfamily 1 group H receptors (NR1H, also known as liver X receptors), and reducing the expression of the genes encoding NR1H3 or NR1H2 increased stromal cell tolerance to pyolysin. In conclusion, mevalonate-derived isoprenoids increased bovine endometrial stromal cell tolerance to pyolysin, which was associated with reducing cellular cholesterol and inhibiting NR1H receptors.

Bovine postpartum diseases remain one of the most significant and highly prevalent illnesses with negative effects on the productivity, survival, and welfare of dairy cows. Antibiotics are generally considered beneficial in the treatment of endometritis; however, frequent usage of each antibiotic drug is reason for the emergence of multidrug resistance (MDR) of the pathogenic microorganisms, representing a major impediment for the successful diagnosis and management of infectious diseases in both humans and animals. We synthesized silver nanoparticles (AgNPs) with an average size of 10 nm using the novel biomolecule apigenin as a reducing and stabilizing agent, and evaluated the efficacy of the AgNPs on the MDR pathogenic bacteria Prevotella melaninogenica and Arcanobacterium pyogenes isolated from uterine secretion samples. AgNPs inhibited cell viability and biofilm formation in a dose- and time-dependent manner. Moreover, the metabolic toxicity of the AgNPs was assessed through various cellular assays. The major toxic effect of cell death was caused by an increase in oxidative stress, as evidenced by the increased generation of reactive oxygen species (ROS), malondialdehyde, protein carbonyl content, and nitric oxide. The formation of ROS is considered to be the primary mechanism of bacterial death. Therefore, the biomolecule-mediated synthesis of AgNPs shows potential as an alternative antimicrobial therapy for bovine metritis and endometritis.

Trueperella pyogenes is an opportunistic pathogen that causes suppurative infections in animals including humans. Data on phenotypic and genotypic properties of T. pyogenes isolated from ruminants, particularly goats and sheep, are lacking. We characterized, by phenotypic and genotypic means, T. pyogenes of caprine and ovine origin, and established their phylogenetic relationship with isolates from other ruminants. T. pyogenes isolates ( n = 50) from diagnostic specimens of bovine ( n = 25), caprine ( n = 19), and ovine ( n = 6) origin were analyzed. Overall, variable biochemical activities were observed among the T. pyogenes isolates. The fimbriae-encoding gene, fimE, and neuraminidase-encoding gene, nanH, were, respectively, more frequently detected in the large ( p = 0.0006) and small ( p = 0.0001) ruminant isolates. Moreover, genotype V ( plo/ nanH/ nanP/ fimA/ fimC) was only detected in the caprine and ovine isolates, whereas genotype IX ( plo/ nanP/ fimA/ fimC/ fimE) was solely present in the isolates of bovine origin ( p = 0.0223). The 16S rRNA gene sequences of all T. pyogenes isolates were clustered with the reference T. pyogenes strain ATCC 19411 and displayed a high degree of identity to each other. Our results highlight phenotypic and genotypic diversity among ruminant isolates of T. pyogenes and reinforce the importance of characterization of more clinical isolates to better understand the pathogenesis of this bacterium in different animal species.

The newly described type strains Arcanobacterium pinnipediorum DSM 28752

There is currently only limited information on the antimicrobial susceptibility and resistance of

There is currently only limited information on the antimicrobial susceptibility and resistance of Corynebacterium spp., Arcanobacterium spp., and Trueperella pyogenes from animals. The comparability of the data is hampered by the use of different antimicrobial susceptibility testing methods and interpretive criteria. To date, standard broth microdilution methods and clinical breakpoints that are approved by the Clinical and Laboratory Standards Institute and are applicable to Corynebacterium spp., Arcanobacterium spp., and T. pyogenes are available. The lack of species-specific clinical breakpoints for the different animal species reduces the explanatory power of the data. Among the isolates of the three genera, elevated MICs for different classes of antimicrobial agents (e.g., β-lactams, macrolides, lincosamides, tetracyclines, aminoglycosides, phenicols, sulfonamides/diaminopyrimidines, and fluoroquinolones) have been described. The most comprehensive data set is available for T. pyogenes , which also includes information about genes and mutations involved in antimicrobial resistance. In T. pyogenes isolates, the macrolide-lincosamide-streptogramin B resistance genes erm (B) and erm (X) were identified. Tetracycline resistance in T. pyogenes was based on the resistance genes tet (W), tet (Z), and tet (33), whereas the aminoglycoside resistance genes aacC , aadA1 , aadA2 , aadA5 , aadA24 , and aadB have been described in T. pyogenes . So far, only single genes conferring either phenicol resistance ( cmlA6 ), trimethoprim resistance ( dfrB2a ), or β-lactam resistance ( blaP1 ) are known to occur in T. pyogenes isolates. Various 23S rRNA mutations, including A2058T, A2058G, and G2137C, were identified in macrolide/lincosamide-resistant T. pyogenes .

Bacteriological examinations were conducted for seven Arcanobacterium haemolyticum strains isolated from elderly patients with skin and soft-tissue infections, such as cellulitis and skin ulcers. Streptococcus dysgalactiae or Gram-positive cocci were isolated together with A. haemolyticum from all patients. The strains were identified as A. haemolyticum based on their being catalase negative, reverse Christie, Atkins and Munch-Petersen (CAMP) positive and phospholipase D gene positive in respective tests. Moreover, API Coryne and matrix-assisted laser desorption/ionization time-of-flight mass spectrometry confirmed the identification of A. haemolyticum. All strains showed good susceptibility to minocycline, vancomycin and β-lactam antibiotics, but several strains were resistant to gentamicin and levofloxacin.