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Staphylococcus schleiferi and Staphylococcus coagulans, closely related bacterial species within the Staphylococcus genus, present a challenge in classification and diagnosis due to their close genetic proximity and overlapping phenotypic features. Moreover, our understanding of the virulence mechanisms in staphylococcal species, beyond the extensively studied Staphylococcus aureus, remains limited, underscoring the importance of using comparative data to enhance our insights into virulence within these bacterial species. This study employed a comprehensive approach, utilizing comparative genomics, to identify genomic distinctions between S. schleiferi and S. coagulans, aiming to address the challenges in the accurate classification and diagnosis of these organisms and identify unique features. Whole genome sequencing was performed on six clinical isolates, and their genomes were compared to identify variations in gene content and virulence factors. De novo assembly and annotation revealed two samples as S. coagulans and four samples as S. schleiferi. Analysis of the core genomes revealed conserved regions crucial for defining species identity, while accessory genomic elements contained unique genes, possibly impacting the pathogenicity of the species.
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Enfermedades de los Perros , Infecciones Cutáneas Estafilocócicas , Animales , Perros , Infecciones Cutáneas Estafilocócicas/tratamiento farmacológico , Infecciones Cutáneas Estafilocócicas/microbiología , Antibacterianos/uso terapéutico , Staphylococcus/genética , Genómica , Secuenciación Completa del GenomaRESUMEN
Parelaphostrongylus tenuis causes ungulate morbidity and mortality in eastern and central North America, but no reference genome sequence exists to facilitate research. Here, we present a P. tenuis genome assembly and annotation, generated with PacBio and Illumina technologies. The assembly is 491 Mbp, with 7285 scaffolds and 185 kb N50.
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Staphylococcus schleiferi and Staphylococcus coagulans are opportunistic pathogens of animals and humans. They were previously classified as Staphylococcus schleiferi subs. schleiferi and Staphylococcus schleiferi subs. coagulans, respectively, and recently reclassified as separate species. S. coagulans, is frequently associated with dogs, whereas S. schleiferi is more commonly isolated from humans. Coagulase activity status is a defining characteristic of the otherwise closely related species. However, the use of coagulase tests originally developed to distinguish S. aureus from non-coagulase-producing staphylococci, for this purpose is questionable and the basis for their host preference has not been elucidated. In the current study, a putative coa gene was identified and correlated with coagulase activity measured using a chromogenic assay with human and bovine prothrombin (closely related to canine prothrombin). The results of the tests performed with human prothrombin showed greater reactivity of S. coagulans isolates from humans than isolates obtained from dogs with the same substrate. Our data suggest that unlike S. coagulans isolates from humans, isolates from dogs have more coagulase activity with bovine prothrombin (similar to canine prothrombin) than human prothrombin. Differences in nuc and 16s rRNA genes suggest a divergence in S. coagulans and S. schleiferi. Phenotypic and genotypic variation based on the number of IgG binding domains, and the numbers of tandem repeats in C-terminal fibronectin binding motifs was also found in protein A, and fibronectin-binding protein B respectively. This study identified a coa gene and associated phenotypic activity that differentiates S. coagulans and S. schleiferi and identified key phylogenetic and phenotypic differences between the species.
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Enfermedades de los Perros , Infecciones Estafilocócicas , Animales , Humanos , Perros , Bovinos , Staphylococcus aureus/genética , Staphylococcus aureus/metabolismo , Coagulasa/genética , Coagulasa/metabolismo , ARN Ribosómico 16S/genética , Fibronectinas/genética , Filogenia , Protrombina , Staphylococcus/metabolismo , Infecciones Estafilocócicas/veterinariaRESUMEN
Staphylococcus pseudintermedius is an important cause of clinical infections in small-animal-veterinary medicine. Evolutionary changes of strains using multilocus sequence typing (MLST) have been observed among S. pseudintermedius in European countries and the United States. However, there are limited or no studies on the detection of methicillin resistant Staphylococcus pseudintermedius (MRSP) and predominating MLST strains in South Africa. Therefore, this study aimed to determine the molecular epidemiology of S. pseudintermedius in South Africa. Twenty-six, non-duplicate, clinical isolates from dogs were obtained as convenience samples from four provinces in South Africa. The Kirby Bauer disk diffusion test was used to determine antimicrobial susceptibility. We used Resfinder and the Comprehensive Antibiotic Resistance Database (CARD) to detect antimicrobial resistance genes. Virulence genes were identified using the virulence factor database and Basic Local Alignment Search Tool (BLASTN) on Geneious prime. geoBURST analysis was used to study relationships between MLST. Finally, the maximum likelihood phylogeny was determined using Randomized Axelerated Maximum Likelihood (RAxML). Twenty-three isolates were confirmed as S. pseudintermedius of which 14 were MRSP. In addition to ß-lactam antimicrobials, MRSP isolates were resistant to tetracycline (85.7%), doxycycline (92.8%), kanamycin (92.8%), and gentamicin (85.7%). The isolates harbored antimicrobial resistance genes (tetM, ermB, drfG, cat, aac(6')-Ie-aph(2")-Ia, ant(6)-Ia, and aph(3')-III) and virulence genes (AdsA, geh, icaA, and lip). MLST analysis showed that ST2228, ST2229, ST2230, ST2231, ST2232, ST2318, ST2326 and ST2327 are unique sequence types in South Africa. Whereas, previously reported major STs including ST45, ST71, ST181, ST551 and ST496 were also detected. The geoBURST and phylogenetic analysis suggests that the isolates in South Africa are likely genetically related to isolates identified in other countries. Highly resistant MRSP strains (ST496, ST71, and ST45) were reported that could present challenges in the treatment of canine infections in South Africa. Hence, we have gained a better understanding of the epidemiology of MRSP in the African continent, the genes involved in resistance and virulence factors associated with these organisms.
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Antibacterianos , Staphylococcus aureus Resistente a Meticilina , Perros , Animales , Epidemiología Molecular , Antibacterianos/farmacología , Sudáfrica/epidemiología , Tipificación de Secuencias Multilocus , Filogenia , Farmacorresistencia Bacteriana/genéticaRESUMEN
The aim of this study was to perform a phenotypic and molecular epidemiological survey to determine temporal changes in the antimicrobial resistance and population structure of methicillin-resistant Staphylococcus pseudintermedius (MRSP) in the United States. Samples from 200 S. pseudintermedius isolates were obtained from veterinary diagnostic facilities located in geographic regions sampled approximately ten years ago and compared to samples obtained in 2021. Kirby-Bauer disk diffusion was used to determine antimicrobial susceptibility. geoBURST analysis and MrBayes were used to infer relationships of isolates using MLST data. Almost all MRSP isolates (98%) in 2021 were multidrug-resistant with 21% of these isolates resistant to more than 16 antimicrobials. In 2010, 190 S. pseudintermedius isolates were collected and 141 of them were MRSP. From 2010-2021 there was a significant increase in resistance observed to all antibiotics tested except cephalothin and sulfonamides. Whereas ten years previously multilocus sequence types (ST) ST68 (35.7%), ST71 (10%), and ST84 (17.4%) predominated, these strains have been supplanted by other STs, notably ST45 (n = 14), ST155 (n = 9), ST181 (n = 13), ST496 (n = 9) and ST551 (n = 9). The newly prevalent STs are distantly related to ST68, ST71 and ST84 and most likely do not share any recent common ancestors. The population structure of MRSP is far more elastic than expected with new, highly resistant strains replacing the ones that predominated in the United States a decade ago. Antibiotic use may play a role in selection; however, the strains that were replaced were also multidrug-resistant and other factors are likely involved.
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Antiinfecciosos , Enfermedades de los Perros , Staphylococcus aureus Resistente a Meticilina , Infecciones Estafilocócicas , Perros , Estados Unidos/epidemiología , Animales , Resistencia a la Meticilina , Infecciones Estafilocócicas/epidemiología , Infecciones Estafilocócicas/veterinaria , Tipificación de Secuencias Multilocus/veterinaria , Enfermedades de los Perros/epidemiología , Antibacterianos/farmacología , Pruebas de Sensibilidad Microbiana/veterinariaRESUMEN
Foods that contain antibiotic residues have potential adverse health effects on consumers and provide selective pressure for the threat of antimicrobial resistance (AMR). This study's objective was to measure tetracycline, sulfonamide, and erythromycin residues in beef, eggs, and honey sold as "antibiotic-free" at farmers' markets in East Tennessee (East TN) in the United States (U.S.). Between July and September 2020, 36 "antibiotic-free" food products (9 beef, 18 egg, and 9 honey products) were purchased from East TN farmers' markets and tested for tetracycline, sulfonamide, and erythromycin residues using competitive enzyme-linked immunosorbent assays (cELISA). All beef, egg, and honey products had tetracycline residue; the median concentrations were 51.75, 30.25, and 77.86 µg/kg, respectively. Sulfonamide residue was present in every sample of beef. Of 18 eggs, 11 eggs had detectable sulfonamide residue; the median concentrations were 3.50 and 1.22 µg/kg in beef and eggs, respectively. Each sample of beef and honey contained erythromycin residue; the median concentrations were 3.67 and 0.68 µg/kg, respectively. Overall, the median concentrations of tetracycline, sulfonamide, and erythromycin residues were below the maximum residue levels (MRLs) set in the U.S. for beef and eggs. Thus, the beef and eggs sold as "antibiotic-free" in East TN farmers' markets can be considered safe for consumption. Safety determination for honey could not be made because MRLs have not been set for honey in the U.S. Because these residues should not be expected in "antibiotic-free" food products, it is important to further investigate the potential sources of these residues in these products.
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We report here the genome sequences of 11 canine Staphylococcus pseudintermedius isolates from New York, New Hampshire, California, Pennsylvania, and Kansas. The sequencing information will enable spatial phylogenetic comparisons of staphylococcal species and other related species and will help in better understanding their virulence potential.
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Respiratory syncytial virus (RSV) is the leading cause of respiratory illness in ruminants and infants. The G glycoprotein of RSV serves as the viral attachment ligand. Despite currently available vaccines, RSV immunity is insufficient, and re-infections occur. Vaccine studies employing the G-protein's 174-187 amino acids, representing the immunodominant domain, have protected mice and calves against infections. To investigate the causes of vaccination failure, we designed four synthetic peptides for the ruminant RSV isolates (391-2, Maryland-BRSV, European-BRSV, and ORSV) using the immune-dominant sequence and vaccinated mice groups with them. The produced antibodies targeting each peptide were evaluated using ELISA and flow cytometry to determine their reactivity against the linear antigen and the native form of the G protein, respectively. Antibodies responded to homologous and heterologous peptides as determined by ELISA. Using flow cytometry-analysis targeting the natively folded protein, most generated antibodies reacted only with their homologous strain. However, antibodies raised to 391-2 peptide reacted with homologous and heterologous Maryland-BRSV viral epitopes. Accordingly, inadequate immunity and recurring RSV infections might be attributed to variations of antibodies targeting the immunodominant region of the G-protein.
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Infecciones por Virus Sincitial Respiratorio , Virus Sincitial Respiratorio Humano , Bovinos , Animales , Ratones , Epítopos Inmunodominantes , Ratones Endogámicos BALB C , Aminoácidos , Formación de Anticuerpos , Anticuerpos Antivirales , Proteínas de Unión al GTPRESUMEN
Background: Understanding drivers of multidrug resistance (MDR) and methicillin resistance, which have increased among canine staphylococcal isolates, is essential for guiding antimicrobial use practices. Therefore, the objective of this study was to identify predictors of MDR and methicillin resistance among Staphylococcus spp. commonly isolated from canine clinical specimens. Methods: This retrospective study used records of canine specimens submitted to the University of Tennessee College of Veterinary Medicine Clinical Bacteriology Laboratory for bacterial culture and antimicrobial susceptibility testing between 2006 and 2017. Records from 7,805 specimens positive for the following Staphylococcus species were included for analysis: Staphylococcus pseudintermedius, Staphylococcus aureus, Staphylococcus coagulans (formerly Staphylococcus schleiferi subspecies coagulans), and Staphylococcus schleiferi (formerly S. schleiferi subsp. schleiferi). Generalized linear regression models were fit using generalized estimating equations (GEE) to identify predictors of MDR (defined as resistance to three or more antimicrobial classes) and methicillin resistance among these isolates. Results: Multidrug resistance (42.1%) and methicillin resistance (31.8%) were relatively common. Isolates from skeletal (joint and bone) specimens had the highest levels of MDR (51.3%) and methicillin resistance (43.6%), followed by cutaneous specimens (45.8% multidrug-resistant, 37.1% methicillin resistant). Staphylococcus species, specimen site, and clinical setting were significant (p < 0.01) predictors of both outcomes. Compared to S. pseudintermedius, S. schleiferi had higher odds of methicillin resistance, while S. coagulans and S. schleiferi had lower odds of MDR. The odds of both MDR and methicillin resistance for isolates from hospital patient specimens were significantly higher than those from referral patients for urine/bladder and otic specimens. Odds of MDR among isolates from skeletal specimens of hospital patients were also higher than those of referral patients. Conclusions: Staphylococcus isolates in this study had substantial levels of MDR and methicillin resistance. Differences in the odds of these outcomes between referral and hospital patient isolates did not persist for all specimen sites, which may reflect differences in diagnostic testing and antimicrobial use practices with respect to body site or system. Judicious antimicrobial use, informed by culture and susceptibility testing, is important to limit treatment failures and curb selection pressure.
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Antibacterianos , Resistencia a la Meticilina , Animales , Perros , Antibacterianos/farmacología , Estudios Retrospectivos , Tennessee/epidemiología , Staphylococcus , Resistencia a Múltiples MedicamentosAsunto(s)
Dermatitis , Enfermedades de los Caballos , Animales , Dermatitis/tratamiento farmacológico , Dermatitis/epidemiología , Dermatitis/prevención & control , Dermatitis/veterinaria , Ácidos Grasos , Enfermedades de los Caballos/tratamiento farmacológico , Enfermedades de los Caballos/epidemiología , Enfermedades de los Caballos/prevención & control , Caballos , Esfingosina/análogos & derivadosRESUMEN
In late 2019, a novel coronavirus began circulating within humans in central China. It was designated SARS-CoV-2 because of its genetic similarities to the 2003 SARS coronavirus (SARS-CoV). Now that SARS-CoV-2 has spread worldwide, there is a risk of it establishing new animal reservoirs and recombination with native circulating coronaviruses. To screen local animal populations in the United States for exposure to SARS-like coronaviruses, we developed a serological assay using the receptor binding domain (RBD) from SARS-CoV-2. SARS-CoV-2's RBD is antigenically distinct from common human and animal coronaviruses, allowing us to identify animals previously infected with SARS-CoV or SARS-CoV-2. Using an indirect enzyme-linked immunosorbent assay (ELISA) for SARS-CoV-2's RBD, we screened serum from wild and domestic animals for the presence of antibodies against SARS-CoV-2's RBD. Surprisingly prepandemic feline serum samples submitted to the University of Tennessee Veterinary Hospital were â¼50% positive for anti-SARS RBD antibodies. Some of these samples were serologically negative for feline coronavirus (FCoV), raising the question of the etiological agent generating anti-SARS-CoV-2 RBD cross-reactivity. We also identified several white-tailed deer from South Carolina with anti-SARS-CoV-2 antibodies. These results are intriguing, as cross-reactive antibodies toward SARS-CoV-2 RBD have not been reported to date. The etiological agent responsible for seropositivity was not readily apparent, but finding seropositive cats prior to the current SARS-CoV-2 pandemic highlights our lack of information about circulating coronaviruses in other species. IMPORTANCE We report cross-reactive antibodies from prepandemic cats and postpandemic South Carolina white-tailed deer that are specific for that SARS-CoV RBD. There are several potential explanations for this cross-reactivity, each with important implications to coronavirus disease surveillance. Perhaps the most intriguing possibility is the existence and transmission of an etiological agent (such as another coronavirus) with similarity to SARS-CoV-2's RBD region. However, we lack conclusive evidence of prepandemic transmission of a SARS-like virus. Our findings provide impetus for the adoption of a One Health Initiative focusing on infectious disease surveillance of multiple animal species to predict the next zoonotic transmission to humans and future pandemics.
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Anticuerpos Antivirales , Gatos , Ciervos , Glicoproteína de la Espiga del Coronavirus , Animales , Anticuerpos Neutralizantes/inmunología , Anticuerpos Antivirales/inmunología , COVID-19/diagnóstico , COVID-19/veterinaria , Gatos/virología , Reacciones Cruzadas/inmunología , Ciervos/virología , Coronavirus Relacionado al Síndrome Respiratorio Agudo Severo/inmunología , SARS-CoV-2 , Glicoproteína de la Espiga del Coronavirus/metabolismo , Zoonosis Virales/diagnóstico , Zoonosis Virales/virologíaRESUMEN
BACKGROUND: Multidrug- and methicillin-resistant staphylococci are both veterinary and public health concerns due to their zoonotic potential. Therefore, the objective of this study was to investigate patterns of antimicrobial, multidrug, and methicillin resistance among four Staphylococcus spp. commonly isolated from canine clinical specimens submitted to the Clinical Bacteriology Laboratory at the University of Tennessee College of Veterinary Medicine (UTCVM). METHODS: Results of antimicrobial susceptibility testing and mecA polymerase chain reaction (PCR) for isolates of four common Staphylococcus spp. isolates were obtained from the Bacteriology Laboratory at the UTCVM between 01/01/2006 and 12/31/2017. Cochran-Armitage trend test was used to assess temporal trends of antimicrobial resistance (AMR), multidrug resistance (MDR), and methicillin resistance. Kappa test of agreement was used to assess agreement between the results of PCR and disk diffusion tests. RESULTS: Most of the 7805 isolates were S. pseudintermedius (6453 isolates), followed by S. coagulans (860), S. aureus (330), and S. schleiferi (162). Among S. pseudintermedius isolates, 45.5% were MDR, and 30.8% were methicillin-resistant (MRSP). There was a significant temporal increase in MRSP (p = 0.017). Chloramphenicol resistance increased among both MRSP and methicillin-susceptible (MSSP) isolates (p < 0.0001). Among S. aureus isolates, 40.9% were MDR, 37.4% were methicillin-resistant (MRSA), and the proportion of MRSA isolates increased significantly (p = 0.0480) over time. There was an increasing temporal trend in the proportion of MDR isolates among MSSP (p = 0.0022), but a decrease among MRSP (p < 0.0001) and MRSA (p = 0.0298). S. schleiferi had the highest percentage (56.9%) of methicillin-resistant isolates. Oxacillin disk diffusion was superior to cefoxitin for the detection of mecA-mediated resistance and had almost perfect agreement with mecA PCR assay for S. pseudintermedius (95.4% agreement, kappa (κ) = 0.904; p < 0.0001), S. coagulans (95.6%, κ = 0.913; p < 0.0001) and S. schleiferi (97.7%, κ = 0.945; p < 0.0001). However, cefoxitin disk diffusion was superior to oxacillin disk diffusion and had almost perfect agreement with mecA PCR assay for S. aureus (95.3%, κ = 0.834; p < 0.0001). CONCLUSIONS: The levels of resistance and increasing temporal trends are concerning. These findings have implications for treatment decisions and public health due to the zoonotic potential of staphylococci. Continued surveillance and use of antibiograms to guide clinical decisions will be critical.
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Antiinfecciosos , Enfermedades de los Perros , Infecciones Estafilocócicas , Animales , Antibacterianos/farmacología , Enfermedades de los Perros/tratamiento farmacológico , Enfermedades de los Perros/epidemiología , Enfermedades de los Perros/microbiología , Perros , Humanos , Resistencia a la Meticilina , Pruebas de Sensibilidad Microbiana/veterinaria , Infecciones Estafilocócicas/tratamiento farmacológico , Infecciones Estafilocócicas/epidemiología , Infecciones Estafilocócicas/veterinaria , Staphylococcus , Staphylococcus aureus , Tennessee/epidemiologíaRESUMEN
BACKGROUND: Mycobacteria are found in many environmental conditions and infect a variety of species, including rodents and rabbits. Guinea pigs are used experimentally as a model for Mycobacterium tuberculosis, but natural mycobacteriosis in guinea pigs has not been reported. CASE PRESENTATION: A 1.5-year-old female guinea pig was found acutely deceased with no premonitory illness. On gross post-mortem examination, multifocal to coalescing, raised, firm, pale tan nodules with discrete, irregular margins were noted over the surfaces of all lung lobes. Histopathology revealed nodules composed of clustered foamy macrophages and multinucleated giant cells containing numerous bacterial rods. Similar bacteria-laden macrophages were noted within sections of the liver, heart, palpebral conjunctiva, duodenum, and cecum. Polymerase chain reaction was performed on tissues collected during post-mortem examination. The 16S rRNA gene product was sequenced and was identical to the Mycobacterium genavense type strain. CONCLUSIONS: To the best of the author's knowledge, this report details the first documented case of Mycobacterium genvaense infection in a guinea pig and a follow up investigation of close-contact animals. Given their experimental susceptibility and this clinical case report, mycobacteriosis should be considered as a differential in guinea pigs exhibiting weight loss in the absence of other clinical signs. With the potential for zoonotic transmission in immunosuppressed individuals, precautions should be taken to safeguard human health in cases of guinea pigs with suspected M. genavense infection.
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Infecciones por Mycobacterium no Tuberculosas , Mycobacterium , Animales , Femenino , Cobayas , Infecciones por Mycobacterium no Tuberculosas/veterinaria , Reacción en Cadena de la Polimerasa/veterinaria , ARN Ribosómico 16S/genética , ConejosRESUMEN
Staphylococcus pseudintermedius is a major opportunistic bacterial pathogen and the leading cause of pyoderma in dogs. In canines it is also often associated with infections of the urinary system and wounds and occasionally infects people. Widespread antimicrobial resistance has made the development of alternative treatments a high priority. The development of a staphylococcal vaccine, however, has proven challenging. Identification of virulence factors that inhibit phagocytosis and avoid innate immunity may play a significant role in preventing or treating infection with S. pseudintermedius. In this study, we identified a putative 5'-nucleotidase provisionally named SpAdsA, a S. pseudintermedius cell- wall protein encoded by SpAdsA. SpAdsA shares approximately 52% identity with the orthologous protein of Staphylococcus aureus and 14.8% identity with that of Streptococcus suis type2. It catalyzes the dephosphorylation of adenosine triphosphate and attenuation of this enzyme with critical amino acid substitutions nearly eliminated its hydrolytic activity. Exogenous adenosine inhibited phagocytosis of S. pseudintermedius by canine neutrophils and monocytes. Conversely, the addition of SpAdsA inhibitor or A2A adenosine receptor antagonist impaired the capacity of S. pseudintermedius to escape from killing by phagocytic cells. The neutralizing ability of canine antibody produced against SpAdsA-M was determined. Taken together, these results suggest that SpAdsA likely plays an important role in S. pseudintermedius virulence and that attenuated SpAdsA may be a good candidate for inclusion in a vaccine against S. pseudintermedius.
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5'-Nucleotidasa/inmunología , Fagocitos/microbiología , Fagocitosis , Staphylococcus/enzimología , 5'-Nucleotidasa/genética , Adenosina/farmacología , Adenosina Trifosfato/metabolismo , Animales , Proteínas Bacterianas/genética , Proteínas Bacterianas/inmunología , Pared Celular/química , Perros , Fagocitos/inmunología , Fosforilación , Staphylococcus/genética , Factores de VirulenciaRESUMEN
Six Staphylococcus strains were isolated from healthy black bears (Ursus americanus) in the Great Smoky Mountains National Park, Tennessee, USA. Phylogenetic analysis based on complete genome, 16S rRNA, dnaJ, hsp60, rpoB and sodA genes, and MALDI-TOF-MS main spectral profiles revealed that the strains belonged to one species and showed the closest relatedness to members of the 'Staphylococcus intermedius group' (SIG), which include Staphylococcus intermedius, Staphylococcus pseudintermedius, Staphylococcus delphini and Staphyloccoccus cornubiensis. The strains were positive in SIG-specific and negative in individual species-specific PCR assays for the nuc gene. The strains can be differentiated from the other SIG species by the absence of sucrose fermentation, from S. intermedius DSM 20373T, S. pseudintermedius CCUG 49543T and S. cornubiensis DSM 105366T by the absence of methyl ß-d-glucopyranoside fermentation and from S. delphini DSM 20771T by fermentation of trehalose. DNA relatedness of the type strain MI 10-1553T with the type strains of S. delphini, S. pseudintermedius, S. intermedius and S. cornubiensis was ≤48.2â% by digital DNA-DNA hybridization and ≤92.3â% by average nucleotide identity calculations. Iso-C15:0, anteiso-C15â:â0 and anteiso-C17â:â0 were the most common fatty acids. Polar lipids consisted of phosphadidylglycerols, phospholipids, glycolipid, diphosphatidylglycerol and aminophospholipid. Cell-wall peptidoglycan was of type A3α l-Lys-Gly3 (Ser; similar to A11.2 and A11.3). The respiratory quinone belonged to menaquinone 7 (MK-7). The G+C content of MI 10-1553T was 39.3 mol%. The isolated strains represent a novel species of the genus Staphylococcus, for which we propose the name Staphylococcus ursi sp. nov. The type strain is MI 10-1553T (=ATCC TSD-55T=CCOS 1900T).
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Filogenia , Staphylococcus/clasificación , Ursidae/microbiología , Animales , Técnicas de Tipificación Bacteriana , Composición de Base , ADN Bacteriano/genética , Ácidos Grasos/química , Genes Bacterianos , Hibridación de Ácido Nucleico , Peptidoglicano/química , Fosfolípidos/química , ARN Ribosómico 16S/genética , Análisis de Secuencia de ADN , Staphylococcus/aislamiento & purificación , Staphylococcus intermedius/genética , Tennessee , Vitamina K 2/análogos & derivados , Vitamina K 2/químicaRESUMEN
The success of staphylococci as pathogens has been attributed, in part, to their ability to evade their hosts' immune systems. Although the proteins involved in evasion have been extensively studied in staphylococci affecting humans little characterization has been done with Staphylococcus pseudintermedius, an important cause of pyoderma in dogs. Staphylococcus aureus binder of immunoglobulin (Sbi) interferes with innate immune recognition by interacting with multiple host proteins. In this study, a S. pseudintermedius gene that shares 38% similarity to S. aureus Sbi was cloned from S. pseudintermedius strains representative of major clonal lineages bearing two paralogs of the protein. Binding of immunoglobulins and Fab and Fc fragments as well as interaction with complement was measured. S. pseudintermedius Sbi protein bound IgG from multiple species and canine complement C3, neutralized complement activity and bound to canine IgM and B cells. Evidence from this work suggests Sbi may play an important role in S. pseudintermedius immune evasion.
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Proteínas Bacterianas/genética , Proteínas Portadoras/genética , Proteínas del Sistema Complemento/inmunología , Fragmentos Fab de Inmunoglobulinas/inmunología , Fragmentos Fc de Inmunoglobulinas/inmunología , Inmunoglobulina M/inmunología , Staphylococcus/inmunología , Animales , Proteínas Bacterianas/inmunología , Proteínas Portadoras/inmunología , Perros , Homología de Secuencia de Aminoácido , Staphylococcus/genéticaRESUMEN
Staphylococci have evolved numerous strategies to evade their hosts' immune systems. Some staphylococcal toxins target essential components of host innate immunity, one of the two main branches of the immune system. Analysis of the Staphylococcus pseudintermedius secretome using liquid chromatography mass spectrometry guided by genomic data, was used to identify an S. pseudintermedius exotoxin provisionally named SpEX. This exoprotein has low overall amino acid identity with the Staphylococcus aureus group of proteins named staphylococcal superantigen like proteins (SSLs) and staphylococcal enterotoxin- like toxin X (SEIX), but predictive modeling showed that it shares similar folds and domain architecture to these important virulence factors. In this study, we found SpEX binds to complement component C5, prevents complement mediated lysis of sensitized bovine red blood cells, kills polymorphonuclear leukocytes and monocytes and inhibits neutrophil migration at sub-lethal concentrations. A mutant version of SpEX, produced through amino acid substitution at selected positions, had diminished cytotoxicity. Anti-SpEX produced in dogs reduced the inhibitory effect of native SpEX on canine neutrophil migration and protected immune cells from the toxic effects of the native recombinant protein. These results suggest that SpEX likely plays an important role in S. pseudintermedius virulence and that attenuated SpEX may be an important candidate for inclusion in a vaccine against S. pseudintermedius infections.
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Clonación Molecular/métodos , Exotoxinas/genética , Exotoxinas/metabolismo , Staphylococcus/patogenicidad , Sustitución de Aminoácidos , Animales , Toxinas Bacterianas/genética , Toxinas Bacterianas/metabolismo , Bovinos , Cromatografía Liquida , Complemento C5/metabolismo , Perros , Exotoxinas/química , Exotoxinas/toxicidad , Espectrometría de Masas , Modelos Moleculares , Dominios Proteicos , Staphylococcus/genética , Staphylococcus/metabolismoRESUMEN
The Staphylococcus intermedius group is a collection of coagulase-positive staphylococci composed of 5 members, including Staphylococcus pseudintermedius, a zoonotic pathogen often associated with exposure to dogs, and Staphylococcus delphini, which has not previously been recovered from humans. Here, we describe the first human case of S. delphini infection.