RESUMEN

Antimicrobial resistance (AMR) is global health concern escalating rapidly in both clinical settings and environment. The effluent from pharmaceuticals and hospitals may contain diverse antibiotics, exerting selective pressure to develop AMR. To study the aquatic prevalence of drug-resistant staphylococci, sampling was done from river Yamuna (3 sites) and wastewater (7 sites) near pharmaceutical industries in Delhi-NCR, India. 59.25% (224/378) were considered presumptive staphylococci while, methicillin resistance was noted in 25% (56/224) isolates. Further, 23 methicillin-resistant coagulase negative staphylococci (MR-CoNS) of 8 different species were identified via 16S rRNA gene sequencing. Multidrug resistance (MDR) was noted in 60.87% (14/23) isolates. PCR based detection of antibiotic resistance genes revealed the number of isolates containing mecA (7/23), blaZ (6/23), msrA (10/23), aac(6')aph (2") (2/23), aph(3')-IIIa (2/23), ant(4')-Ia (1/23), dfrG (4/23), dfrA(drfS1) (3/23), tetK (1/23) and tetM (1/23). The current research highlights the concerning prevalence of MDR-CoNS in aquatic environment in Delhi.

Asunto(s)

Antibacterianos , Coagulasa , Farmacorresistencia Bacteriana Múltiple , ARN Ribosómico 16S , Staphylococcus , Aguas Residuales , India/epidemiología , Aguas Residuales/microbiología , Staphylococcus/genética , Staphylococcus/efectos de los fármacos , Staphylococcus/aislamiento & purificación , Staphylococcus/clasificación , Farmacorresistencia Bacteriana Múltiple/genética , Coagulasa/metabolismo , Coagulasa/genética , ARN Ribosómico 16S/genética , Antibacterianos/farmacología , Prevalencia , Pruebas de Sensibilidad Microbiana

RESUMEN

The Staphylococcus intermedius group (SIG) includes coagulase-positive staphylococci commonly found in animals. The taxonomic classification within the SIG has evolved with molecular techniques distinguishing five species. Despite their similarities, these species exhibit varied host affinities, with unclear implications for virulence and host interaction. This study aimed to investigate the presence of coagulase-positive staphylococci in pigeons and to detect genes encoding for selected virulence factors in isolated strains. Another goal was to determine the adhesion capabilities of randomly selected pigeon S. intermedius, S. delphini, and canine S. pseudintermedius strains to canine and pigeon corneocytes and their adhesion and invasion abilities to canine keratinocytes in vitro. In total, 121 coagulase-positive strains were isolated from domestic and feral pigeons. The most prevalent species were S. delphini B and S. intermedius in domestic and feral pigeons, respectively. We proved that pigeon strains carried genes encoding for exfoliative toxin SIET and leukotoxin Luk-I. Moreover, we found that S. intermedius showed higher adherence to pigeon than to canine corneocytes, aligning with its presumed natural host. No difference in adherence abilities of S. pseudintermedius to canine and pigeon corneocytes was observed. In this study, we also observed that S. pseudintermedius could successfully invade the canine keratinocytes, in contrary to S. delphini and S. intermedius. Moreover, only S. intermedius was not able to invade canine keratinocytes at all. These findings highlight the complex interplay between SIG bacteria, and their hosts, underscoring the need for further research to understand the mechanisms of host adaptation and pathogenicity within this group.

Asunto(s)

Adhesión Bacteriana , Columbidae , Especificidad del Huésped , Queratinocitos , Infecciones Estafilocócicas , Staphylococcus intermedius , Staphylococcus , Factores de Virulencia , Animales , Columbidae/microbiología , Perros , Factores de Virulencia/genética , Staphylococcus/genética , Staphylococcus/patogenicidad , Staphylococcus/clasificación , Staphylococcus/aislamiento & purificación , Queratinocitos/microbiología , Virulencia/genética , Infecciones Estafilocócicas/microbiología , Infecciones Estafilocócicas/veterinaria , Staphylococcus intermedius/genética , Staphylococcus intermedius/patogenicidad , Coagulasa/metabolismo , Coagulasa/genética , Exfoliatinas/genética , Exfoliatinas/metabolismo , Proteínas Bacterianas/genética , Proteínas Bacterianas/metabolismo

RESUMEN

BACKGROUND: Several diagnostic environments in Uganda lack real-time, robust and high-throughput technologies for comprehensive typing of microbes, which is a setback to infectious disease surveillance. This study combined various wet laboratory diagnostics to understand the epidemiology of pathogenic staphylococci isolated from animals in Uganda and the implications for global health security priorities. METHODS: A retrospective study was conducted employing records and pathogenic staphylococci (from animals) archived at the Central Diagnostic Laboratory (CDL), Makerere University, Uganda, between January 2012 and December 2019. The bacteria were speciated by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) and tested for virulence factors [beta lactamases, lecithinase, deoxyribonuclease (DNase), haemolysins] and resistance to ten antimicrobials of clinical and veterinary relevance. Tetracycline and methicillin resistance genes were also tested. RESULTS: The prevalent diseases were mastitis in cattle and skin infections in dogs. Of the 111 staphylococci tested by MALDI-TOF MS, 79 (71.2%) were Staphylococcus aureus, 27 (24.3%) were Staphylococcus pseudintermedius and 5 (4.5%) were Staphylococcus schleiferi. All these strains expressed haemolysins. The prevalence of strains with lecithinase, penicillinase, cephalosporinase and DNase was 35.9% (14/39), 89.7% (35/39), 0.0% (0/39) and 87.2% (34/39), respectively. Staphylococci were primarily resistant to early penicillins (over 80%), tetracycline (57.7%), and chloramphenicol (46.2%). Minimal resistance was noted with cloxacillin (0.0%), ciprofloxacin (9.6%), and cefoxitin (3.8%). The prevalence of multidrug resistance (MDR) was 78.8% for general staphylococci, 82.2% for S. aureus, 73.1% for S. pseudintermedius, and 60.0% for S. schleiferi. Multidrug resistant staphylococci were significantly more prevalent in the cattle isolates than in the dog isolates (P < 0.05). The prevalence of methicillin-resistant staphylococci (MRS) tested by resistance to cefoxitin and mecA carriage was 3.8%. These four strains were all isolated from dog skin infections. The tetK gene was the most predominant (35.4%), followed by tetM (25.0%). CONCLUSION: In resource-constrained settings, the approach of integrated diagnostics promises sustainable disease surveillance and the addressing of current capacity gaps. The emergence of MRS (zoonotic bacteria) in companion animals creates a likelihood of reduced treatment options for related human infections, a threat to global health.

Asunto(s)

Infecciones Estafilocócicas , Staphylococcus , Animales , Uganda/epidemiología , Infecciones Estafilocócicas/microbiología , Infecciones Estafilocócicas/veterinaria , Infecciones Estafilocócicas/epidemiología , Bovinos , Estudios Retrospectivos , Staphylococcus/genética , Staphylococcus/efectos de los fármacos , Staphylococcus/aislamiento & purificación , Staphylococcus/clasificación , Perros , Espectrometría de Masa por Láser de Matriz Asistida de Ionización Desorción/métodos , Antibacterianos/farmacología , Factores de Virulencia/genética , Femenino , Enfermedades de los Perros/microbiología , Enfermedades de los Perros/epidemiología , Enfermedades de los Perros/diagnóstico , Enfermedades de los Bovinos/microbiología , Enfermedades de los Bovinos/epidemiología , Enfermedades de los Bovinos/diagnóstico , Pruebas de Sensibilidad Microbiana

RESUMEN

Staphylococcus argenteus is a novel species within the Staphylococcus aureus complex and can cause serious bloodstream infections (BSIs) in humans, which have been mainly reported in adults, especially the elderly. In this study, we analyzed the molecular characterization of a strain of S. argenteus (22WJ8192) isolated from the peripheral vein blood sample of a seven-month-old female infant in Eastern China. The 22WJ8192 belonged to sequence type (ST)2250 and harbored six antibiotic-resistance genes and 53 virulence genes and was resistant to penicillin. Additionally, we conducted a comparative analysis of the molecular characteristics of S. argenteus sourced from various origins within the dataset, predominantly from the National Center for Biotechnology Information Collection (NCBI) genome database. Antibiotic-resistance genes blaR1, blaI_of_Z, blaZ, fosB-Saur, tet(L), aph(3")-IIIa, mecA, and dfrG were more prevalent among the strains of human origin. Virulence genes lukF-PV, sak, sdrE, scn, sdrC, and sdrD were more prevalent among strains of human origin. The presence of antibiotic-resistance genes blaR1, blaI_of_Z, blaZ, fosB-Saur, and aph(3")-IIIa in strain 22WJ8192 was also more common among strains of human origin in the dataset. Conversely, the antibiotic-resistance genes tet(L), mecA, and dfrG, typically found in strains of human origin, were not detected in 22WJ8192. Additionally, virulence genes lukF-PV, sak, sdrE, scn, sdrC, and sdrD present in 22WJ8192 exhibited a higher prevalence among strains of human origin in the dataset. In conclusion, this study emphasizes the potential of S. argenteus ST2250 to induce severe bloodstream infections in infants, shedding light on the molecular characteristics of this strain.

Asunto(s)

Antibacterianos , Infecciones Estafilocócicas , Staphylococcus , Factores de Virulencia , Humanos , China/epidemiología , Infecciones Estafilocócicas/microbiología , Infecciones Estafilocócicas/epidemiología , Femenino , Staphylococcus/genética , Staphylococcus/efectos de los fármacos , Staphylococcus/aislamiento & purificación , Staphylococcus/clasificación , Staphylococcus/patogenicidad , Lactante , Factores de Virulencia/genética , Virulencia/genética , Antibacterianos/farmacología , Pruebas de Sensibilidad Microbiana , Farmacorresistencia Bacteriana/genética , Hospitales , Bacteriemia/microbiología , Bacteriemia/epidemiología

RESUMEN

BACKGROUND: In 2015, Staphylococcus argenteus was reported for the first time as a novel species of the Staphylococcus aureus complex. While S. argenteus has been found in many countries, its presence in Indonesia has not been reported yet. Our aim is to confirm S. argenteus presence in Indonesia, describe its characteristics and analyze its genomic diversity. METHODS: The S. aureus isolates used in this study were collected from patients with skin and soft tissue infections in Indonesia, between July 2009 to February 2010. Randomly selected isolates were recultured from -80 C° stocks and analyzed using matrix-assisted laser desorption/ionization - time of flight (MALDI-TOF). Isolates identified as S. argenteus, S. roterodami, or S. schweitzeri and S. aureus with a low score in the MALDI-TOF analysis were analyzed by a real-time PCR targeting the nucA gene able to identify true S. argenteus. Isolates identified as S. argenteus were further characterized by whole genome sequencing. Vitek®2 (bioMérieux) was used for antimicrobial susceptibility testing. RESULTS: Fifteen isolates were identified as S. argenteus, with the majority belonging to ST2250. Two pairs of isolates proved to be identical by core genome multilocus sequence typing analysis. Most isolates were susceptible to all antibiotics tested, except for seven isolates (46.7 %) that were resistant to benzylpenicillin, and one isolate was resistant to tetracycline (6.7 %). The presence of resistance genes blaZ and tet(45) correlated with these findings. Notably, the sey enterotoxin gene was prevalent in 80 % of the isolates. Other virulence factor genes were less prevalent. Plasmid replicon types in S. argenteus were also known to S. aureus. CONCLUSION: Our study reveals the occurrence of S. argenteus in Indonesia. The diversity within Indonesian S. argenteus matches the global diversity of S. argenteus. Identical isolates between patients indicate potential transmission events. A lower prevalence of a broad panel of virulence factors suggests that S. argenteus is less virulent than S. aureus.

Asunto(s)

Antibacterianos , Pruebas de Sensibilidad Microbiana , Espectrometría de Masa por Láser de Matriz Asistida de Ionización Desorción , Infecciones Estafilocócicas , Staphylococcus , Secuenciación Completa del Genoma , Indonesia/epidemiología , Humanos , Staphylococcus/genética , Staphylococcus/aislamiento & purificación , Staphylococcus/clasificación , Staphylococcus/efectos de los fármacos , Infecciones Estafilocócicas/microbiología , Infecciones Estafilocócicas/epidemiología , Antibacterianos/farmacología , Masculino , Femenino , Adulto , Tipificación de Secuencias Multilocus , Persona de Mediana Edad , Adulto Joven , Infecciones de los Tejidos Blandos/microbiología , Infecciones de los Tejidos Blandos/epidemiología , Genoma Bacteriano/genética , Anciano , Variación Genética , Adolescente , Reacción en Cadena en Tiempo Real de la Polimerasa

RESUMEN

Staphylococcus pseudintermedius, which is part of the skin microbiome of dogs, causes a variety of opportunistic infections. These infections may become more difficult to treat due to the formation of biofilm. The capacity of S. pseudintermedius to form biofilm, as well as the associated genes, has not been elucidated. This study evaluated the production and composition of S. pseudintermedius biofilm. Samples were collected from both infected dogs and asymptomatic dogs. Isolates were identified using mass spectrometry and Multiplex-PCR. Biofilm production and composition were assessed using a quantitative microtiter plate assay. The presence of ica operon genes and sps genes was investigated using conventional PCR. The investigation of Agr type and virulence genes was conducted in silico on 24 sequenced samples. All strains could produce strong biofilms, with most of the isolates presenting a polysaccharide biofilm. 63.6% of the isolates carried the complete ica operon (ADBC). All samples showed the presence of the genes spsK, spsA, and spsL, while the distribution of other genes varied. Agr type III was the most prevalent (52.2%). All sequenced samples carried the cytotoxins hlb, luk-S, luk-F, as well as the exfoliative toxins siet and se_int. No isolate displayed other exfoliative toxins. Only LB1733 presented a set of different enterotoxins (sea, seb, sec_canine, seh, sek, sel, and seq). Our findings suggest that S. pseudintermedius is a strong producer of biofilm and carries virulence genes.

Asunto(s)

Biopelículas , Enfermedades de los Perros , Staphylococcus , Animales , Biopelículas/crecimiento & desarrollo , Perros , Enfermedades de los Perros/microbiología , Virulencia/genética , Staphylococcus/genética , Staphylococcus/aislamiento & purificación , Staphylococcus/patogenicidad , Staphylococcus/clasificación , Staphylococcus/fisiología , Infecciones Estafilocócicas/microbiología , Infecciones Estafilocócicas/veterinaria , Factores de Virulencia/genética , Proteínas Bacterianas/genética , Operón

RESUMEN

During surveillance of Staphylococcus aureus in lesions from patients with atopic dermatitis (AD), we isolated Staphylococcus argenteus, a species registered in 2011 as a new member of the genus Staphylococcus and previously considered a lineage of S. aureus. Genome sequence comparisons between S. argenteus isolates and representative S. aureus clinical isolates from various origins revealed that the S. argenteus genome from AD patients closely resembles that of S. aureus causing skin infections. We previously reported that 17%-22% of S. aureus isolated from skin infections produce staphylococcal enterotoxin Y (SEY), which predominantly induces T-cell proliferation via the T-cell receptor (TCR) Vα pathway. Complete genome sequencing of S. argenteus isolates revealed a gene encoding a protein similar to superantigen SEY, designated as SargEY, on its chromosome. Population structure analysis of S. argenteus revealed that these isolates are ST2250 lineage, which was the only lineage positive for the SEY-like gene among S. argenteus. Recombinant SargEY demonstrated immunological cross-reactivity with anti-SEY serum. SargEY could induce proliferation of human CD4+ and CD8+ T cells, as well as production of TNF-α and IFN-γ. SargEY showed emetic activity in a marmoset monkey model. SargEY and SET (a phylogenetically close but uncharacterized SE) revealed their dependency on TCR Vα in inducing human T-cell proliferation. Additionally, TCR sequencing revealed other previously undescribed Vα repertoires induced by SEH. SargEY and SEY may play roles in exacerbating the respective toxin-producing strains in AD. IMPORTANCE: Staphylococcus aureus is frequently isolated from active lesions of atopic dermatitis (AD) patients. We reported that 17%-22% of S. aureus isolated from AD patients produced a novel superantigen staphylococcal enterotoxin Y (SEY). Unlike many S. aureus superantigens that activate T cells via T-cell receptor (TCR) Vß, SEY activates T cells via TCR Vα and stimulates cytokine secretion. Staphylococcus argenteus was isolated from AD patients during the surveillance for S. aureus. Phylogenetic comparison of the genome indicated that the isolate was very similar to S. aureus causing skin infections. The isolate encoded a SEY-like protein, designated SargEY, which, like SEY, activated T cells via the TCR Vα. ST2250 is the only lineage positive for SargEY gene. ST2250 S. argenteus harboring a superantigen SargEY gene may be a novel staphylococcal clone that infects human skin and is involved in the exacerbation of AD.

Asunto(s)

Dermatitis Atópica , Enterotoxinas , Genoma Bacteriano , Staphylococcus , Superantígenos , Humanos , Dermatitis Atópica/microbiología , Dermatitis Atópica/inmunología , Superantígenos/genética , Superantígenos/inmunología , Staphylococcus/genética , Staphylococcus/inmunología , Staphylococcus/clasificación , Enterotoxinas/genética , Enterotoxinas/inmunología , Animales , Filogenia , Genómica , Secuenciación Completa del Genoma , Infecciones Estafilocócicas/microbiología , Infecciones Estafilocócicas/inmunología

RESUMEN

Non-aureus staphylococci and mammaliicocci (NASM) are the microorganisms most frequently isolated from milk. Given their numerosity and complexity, MALDI-TOF MS is one of the preferred species identification approaches. Nevertheless, reference mass spectra for the novel species Staphylococcus borealis were included only recently in the Bruker Biotyper System (MBT) library, and other species of veterinary interest such as S. rostri are still absent. This work provides an updated picture of the NASM species found in milk, gained by retrospectively analyzing the data relating to 21,864 milk samples, of which 6,278 from clinical mastitis (CM), 4,039 from subclinical mastitis (SCM), and 11,547 from herd survey (HS), with a spectrum library including both species. As a result, S. borealis was the second most frequently isolated NASM (17.07%) after S. chromogenes (39.38%) in all sample types, with a slightly higher percentage in CM (21.84%), followed by SCM (17.65%), and HS (14.38%). S. rostri was also present in all sample types (3.34%), reaching 8.43% of all NASM in SCM and showing a significant association (p < 0.01) with this condition. Based on our findings, the presence of S. borealis and S. rostri in milk and their potential association with mastitis might have been overlooked, possibly due to the difficulties in differentiating these species from other closely related NASM. Our results indicate that S. borealis could be a more frequent contributor to bovine udder infections than previously thought and that S. rostri should also not be underestimated considering its significant association with SCM.

Asunto(s)

Mastitis Bovina , Leche , Espectrometría de Masa por Láser de Matriz Asistida de Ionización Desorción , Staphylococcus , Animales , Leche/microbiología , Bovinos , Espectrometría de Masa por Láser de Matriz Asistida de Ionización Desorción/veterinaria , Espectrometría de Masa por Láser de Matriz Asistida de Ionización Desorción/métodos , Staphylococcus/aislamiento & purificación , Staphylococcus/clasificación , Femenino , Mastitis Bovina/microbiología , Estudios Retrospectivos , Infecciones Estafilocócicas/veterinaria , Infecciones Estafilocócicas/microbiología

RESUMEN

BACKGROUND: Recently, linezolid-resistant staphylococci have become an emerging problem worldwide. Understanding the mechanisms of resistance, molecular epidemiology and transmission of linezolid-resistant CoNS in hospitals is very important. METHODS: The antimicrobial susceptibilities of all isolates were determined by the microdilution method. The resistance mechanisms and molecular characteristics of the strains were determined using whole-genome sequencing and PCR. RESULTS: All the strains were resistant to oxacillin and carried the mecA gene; 13 patients (36.1%) had prior linezolid exposure. Most S. epidermidis and S. hominis isolates were ST22 and ST1, respectively. MLST typing and evolutionary analysis indicated most linezolid-resistant CoNS strains were genetically related. In this study, we revealed that distinct CoNS strains have different mechanisms of linezolid resistance. Among ST22-type S. epidermidis, acquisition of the T2504A and C2534T mutations in the V domain of the 23 S rRNA gene, as well as mutations in the ribosomal proteins L3 (L101V, G152D, and D159Y) and L4 (N158S), were linked to the development of linezolid resistance. In S. cohnii isolates, cfr, S158Y and D159Y mutations in the ribosomal protein L3 were detected. Additionally, emergence of the G2576T mutation and the cfr gene were major causes of linezolid resistance in S. hominis isolates. The cfr gene, G2576T and C2104T mutations, M156T change in L3 protein, and I188S change in L4 protein were found in S. capitis isolates. CONCLUSION: The emergence of linezolid-resistant CoNS in the environment is concerning because it involves clonal dissemination and frequently coexists with various drug resistance mechanisms.

Asunto(s)

Antibacterianos , Linezolid , Pruebas de Sensibilidad Microbiana , Infecciones Estafilocócicas , Centros de Atención Terciaria , Linezolid/farmacología , Humanos , China/epidemiología , Infecciones Estafilocócicas/microbiología , Infecciones Estafilocócicas/epidemiología , Antibacterianos/farmacología , Femenino , Masculino , Persona de Mediana Edad , Tipificación de Secuencias Multilocus , Anciano , Secuenciación Completa del Genoma , Staphylococcus/efectos de los fármacos , Staphylococcus/genética , Staphylococcus/clasificación , Staphylococcus/enzimología , Coagulasa/metabolismo , Coagulasa/genética , ARN Ribosómico 23S/genética , Adulto , Resistencia a la Meticilina/genética , Mutación , Proteínas Bacterianas/genética

RESUMEN

Cutaneous ulcers are common in yaws-endemic areas. Although often attributed to 'Treponema pallidum subsp. pertenue' and Haemophilus ducreyi, quantitative PCR has highlighted a significant proportion of these ulcers are negative for both pathogens and are considered idiopathic. This is a retrospective analysis utilising existing 16S rRNA sequencing data from two independent yaws studies that took place in Ghana and the Solomon Islands. We characterized bacterial diversity in 38 samples to identify potential causative agents for idiopathic cutaneous ulcers. We identified a diverse bacterial profile, including Arcanobacterium haemolyticum, Campylobacter concisus, Corynebacterium diphtheriae, Staphylococcus spp. and Streptococcus pyogenes, consistent with findings from previous cutaneous ulcer microbiome studies. No single bacterial species was universally present across all samples. The most prevalent bacterium, Campylobacter ureolyticus, appeared in 42% of samples, suggesting a multifactorial aetiology for cutaneous ulcers in yaws-endemic areas. This study emphasizes the need for a nuanced understanding of potential causative agents. The findings prompt further exploration into the intricate microbial interactions contributing to idiopathic yaw-like ulcers, guiding future research toward comprehensive diagnostic and therapeutic strategies.

Asunto(s)

Microbiota , ARN Ribosómico 16S , Úlcera Cutánea , Humanos , ARN Ribosómico 16S/genética , Úlcera Cutánea/microbiología , Ghana , Masculino , Buba/microbiología , Buba/diagnóstico , Estudios Retrospectivos , Femenino , Adulto , Bacterias/genética , Bacterias/clasificación , Bacterias/aislamiento & purificación , Melanesia , Persona de Mediana Edad , Staphylococcus/genética , Staphylococcus/aislamiento & purificación , Staphylococcus/clasificación , Streptococcus pyogenes/genética , Streptococcus pyogenes/aislamiento & purificación , Streptococcus pyogenes/clasificación , Arcanobacterium/genética , Arcanobacterium/aislamiento & purificación , Campylobacter/genética , Campylobacter/aislamiento & purificación , Campylobacter/clasificación

RESUMEN

The evolutionary lineage and taxonomy of the Australian dingo is controversial, however recent genomic and gut metagenomic research has suggested that dingoes are evolutionarily distinct from modern dogs. Staphylococcus species are known commensal organisms of dogs and other mammals. In this study we took the opportunity to determine the carriage rate and antimicrobial resistance profiles of Staphylococcus species from 15 captive Australian dingoes. S. pseudintermedius was the only coagulase-positive species recovered, isolated from 6/15 (40%) and 9/13 (69%) of the animals during the 2020 (winter) and 2021 (summer) sampling times, respectively. Twenty-three coagulase-negative isolates were characterised, with S. equorum being the most frequently (20/23, 87%) recovered species. Two isolates of S. equorum had their genomes sequenced to learn more about this species. Antimicrobial resistance amongst both coagulase-positive and -negative isolates was low; with resistance to only 3 of 12 antimicrobials observed: penicillin, erythromycin, and trimethoprim. We have shown that the Australian dingo is a host organism for S. pseudintermedius much like it is in dogs, however the carriage rate was lower than has previously been reported from dogs in Australia.

Asunto(s)

Antibacterianos , Portador Sano , Infecciones Estafilocócicas , Staphylococcus , Animales , Staphylococcus/efectos de los fármacos , Staphylococcus/clasificación , Staphylococcus/genética , Staphylococcus/aislamiento & purificación , Antibacterianos/farmacología , Portador Sano/microbiología , Portador Sano/veterinaria , Infecciones Estafilocócicas/veterinaria , Infecciones Estafilocócicas/microbiología , Infecciones Estafilocócicas/epidemiología , Victoria/epidemiología , Pruebas de Sensibilidad Microbiana , Farmacorresistencia Bacteriana , Perros/microbiología , Canidae/microbiología , Masculino , Femenino

RESUMEN

BACKGROUND: Staphylococcus pseudintermedius is a common opportunistic pathogen of companion dogs and an occasional human pathogen. Treatment is hampered by antimicrobial resistance including methicillin resistance encoded by mecA within the mobile genetic element SCCmec. OBJECTIVES: SCCmec elements are diverse, especially in non-Staphyloccocus aureus staphylococci, and novel variants are likely to be present in S. pseudintermedius. The aim was to characterize the SCCmec elements found in four canine clinical isolates of S. pseudintermedius. MATERIAL AND METHODS: Isolates were whole-genome sequenced and SCCmec elements were assembled, annotated and compared to known SCCmec types. RESULTS AND DISCUSSION: Two novel SSCmec are present in these isolates. SCCmec7017-61515 is characterized by a novel combination of a Class A mec gene complex and a type 5 ccr previously only described in composite SCCmec elements. The other three isolates share a novel composite SCCmec with features of SCCmec types IV and VI. CONCLUSIONS: S. pseudintermedius is a reservoir of novel SSCmec elements that has implications for understanding antimicrobial resistant in veterinary and human medicine.

Asunto(s)

Cromosomas Bacterianos , Enfermedades de los Perros , Resistencia a la Meticilina , Infecciones Estafilocócicas , Staphylococcus , Secuenciación Completa del Genoma , Resistencia a la Meticilina/genética , Staphylococcus/genética , Staphylococcus/efectos de los fármacos , Staphylococcus/clasificación , Staphylococcus/aislamiento & purificación , Animales , Perros , Infecciones Estafilocócicas/microbiología , Infecciones Estafilocócicas/veterinaria , Enfermedades de los Perros/microbiología , Cromosomas Bacterianos/genética , Proteínas Bacterianas/genética , Antibacterianos/farmacología , Pruebas de Sensibilidad Microbiana , Genoma Bacteriano , Variación Genética , Secuencias Repetitivas Esparcidas/genética

RESUMEN

Human cutaneous squamous cell carcinomas (SCCs) and actinic keratoses (AK) display microbial dysbiosis with an enrichment of staphylococcal species, which have been implicated in AK and SCC progression. SCCs are common in both felines and canines and are often diagnosed at late stages leading to high disease morbidity and mortality rates. Although recent studies support the involvement of the skin microbiome in AK and SCC progression in humans, there is no knowledge of this in companion animals. Here, we provide microbiome data for SCC in cats and dogs using culture-independent molecular profiling and show a significant decrease in microbial alpha diversity on SCC lesions compared to normal skin (P ≤ 0.05). Similar to human skin cancer, SCC samples had an elevated abundance of staphylococci relative to normal skin-50% (6/12) had >50% staphylococci, as did 16% (4/25) of perilesional samples. Analysis of Staphylococcus at the species level revealed an enrichment of the pathogenic species Staphylococcus felis in cat SCC samples, a higher prevalence of Staphylococcus pseudintermedius in dogs, and a higher abundance of Staphylococcus aureus compared to normal skin in both companion animals. Additionally, a comparison of previously published human SCC and perilesional samples against the present pet samples revealed that Staphylococcus was the most prevalent genera across human and companion animals for both sample types. Similarities between the microbial profile of human and cat/dog SCC lesions should facilitate future skin cancer research. IMPORTANCE: The progression of precancerous actinic keratosis lesions (AK) to cutaneous squamous cell carcinoma (SCC) is poorly understood in humans and companion animals, despite causing a significant burden of disease. Recent studies have revealed that the microbiota may play a significant role in disease progression. Staphylococcus aureus has been found in high abundance on AK and SCC lesions, where it secretes DNA-damaging toxins, which could potentiate tumorigenesis. Currently, a suitable animal model to investigate this relationship is lacking. Thus, we examined the microbiome of cutaneous SCC in pets, revealing similarities to humans, with increased staphylococci and reduced commensals on SCC lesions and peri-lesional skin compared to normal skin. Two genera that were in abundance in SCC samples have also been found in human oral SCC lesions. These findings suggest the potential suitability of pets as a model for studying microbiome-related skin cancer progression.

Asunto(s)

Carcinoma de Células Escamosas , Enfermedades de los Gatos , Enfermedades de los Perros , Microbiota , Neoplasias Cutáneas , Piel , Staphylococcus , Gatos , Perros , Animales , Carcinoma de Células Escamosas/microbiología , Carcinoma de Células Escamosas/veterinaria , Neoplasias Cutáneas/microbiología , Neoplasias Cutáneas/veterinaria , Neoplasias Cutáneas/patología , Piel/microbiología , Piel/patología , Enfermedades de los Gatos/microbiología , Staphylococcus/aislamiento & purificación , Staphylococcus/genética , Staphylococcus/clasificación , Staphylococcus/patogenicidad , Enfermedades de los Perros/microbiología , Queratosis Actínica/microbiología , Queratosis Actínica/veterinaria , Queratosis Actínica/patología

RESUMEN

The virulence factors, antibiotic resistance patterns, and the associated genetic elements have been investigated in Staphylococcus species. A total of 100 strains has been isolated from clinical samples in the Microbiology Laboratory of Hesperia Hospital, Modena, Italy, and identified as Staphylococcus aureus (65), Staphylococcus epidermidis (24), Staphylococcus hominis (3), Staphylococcus saprophyticus (3), and Staphylococcus warneri (5). All the strains were analyzed to determine phenotypic and genotypic characters, notably the virulence factors, the antibiotics susceptibility, and the genetic determinants. The highest percentage of resistance in Staphylococcus spp. was found for erythromycin and benzylpenicillin (87% and 85%, respectively). All S. aureus, two S. epidermidis (8.3%), and one S. saprophyticus (33.3%) strains were resistant to oxacillin. The methicillin resistance gene (mecA) was detected by polymerase chain reaction (PCR) amplification in 65 S. aureus strains and in 3 coagulase-negative staphylococci (CoNS) (8.6%). With regard to the virulence characteristics, all the S. aureus were positive to all virulence tests, except for slime test. Among the CoNS isolates, 19 (79.1%) S. epidermidis and one (33.3%) S. saprophyticus strains resulted positive for the slime test only. The results obtained are useful for a more in-depth understanding of the function and contribution of S. aureus and CoNS antibiotic resistance and virulence factors to staphylococcal infections. In particular, the production of slime is very important for CoNS, a virulence factor frequently found in infections caused by these strains. Further investigations on the genetic relatedness among strains of different sources will be useful for epidemiological and monitoring purposes and will enable us to develop new strategies to counteract the diffusion of methicillin-resistant S. aureus (MRSA) and CoNS strains not only in clinical field, but also in other related environments.

Asunto(s)

Antibacterianos , Farmacorresistencia Bacteriana , Pruebas de Sensibilidad Microbiana , Infecciones Estafilocócicas , Staphylococcus , Factores de Virulencia , Factores de Virulencia/genética , Infecciones Estafilocócicas/microbiología , Staphylococcus/genética , Staphylococcus/efectos de los fármacos , Staphylococcus/aislamiento & purificación , Staphylococcus/patogenicidad , Staphylococcus/clasificación , Humanos , Antibacterianos/farmacología , Italia , Proteínas Bacterianas/genética

RESUMEN

The phylogenetic tree of the Staphylococcus aureus complex consists of several distinct clades and the majority of human and veterinary S. aureus isolates form one large clade. In addition, two divergent clades have recently been described as separate species. One was named Staphylococcus argenteus, due to the lack of the "golden" pigment staphyloxanthin. The second one is S. schweitzeri, found in humans and animals from Central and West Africa. In late 2021, two additional species, S. roterodami and S. singaporensis, have been described from clinical samples from Southeast Asia. In the present study, isolates and their genome sequences from wild Straw-coloured fruit bats (Eidolon helvum) and a Diamond firetail (Stagonopleura guttata, an estrildid finch) kept in a German aviary are described. The isolates possessed staphyloxanthin genes and were closer related to S. argenteus and S. schweitzeri than to S. aureus. Phylogenetic analysis revealed that they were nearly identical to both, S. roterodami and S. singaporensis. We propose considering the study isolates, the recently described S. roterodami and S. singaporensis as well as some Chinese strains with MLST profiles stored in the PubMLST database as different clonal complexes within one new species. According to the principle of priority we propose it should be named S. roterodami. This species is more widespread than previously believed, being observed in West Africa, Southeast Asia and Southern China. It has a zoonotic connection to bats and has been shown to be capable of causing skin and soft tissue infections in humans. It is positive for staphyloxanthin, and it could be mis-identified as S. aureus (or S. argenteus) using routine procedures. However, it can be identified based on distinct MLST alleles, and "S. aureus" sequence types ST2470, ST3135, ST3952, ST3960, ST3961, ST3963, ST3965, ST3980, ST4014, ST4075, ST4076, ST4185, ST4326, ST4569, ST6105, ST6106, ST6107, ST6108, ST6109, ST6999 and ST7342 belong to this species.

Asunto(s)

Quirópteros , Filogenia , Staphylococcus , Animales , Quirópteros/microbiología , Tipificación de Secuencias Multilocus , Staphylococcus/clasificación , Staphylococcus/aislamiento & purificación

RESUMEN

Coagulase-negative staphylococci (CoNS) are the main pathogens in health care-associated ventriculitis and meningitis (HCAVM). This study aimed to assess antimicrobial susceptibility. Moreover, the treatment and clinical outcome were described. All neurosurgical adults admitted to one of the largest neurosurgical centers in China with clinically significant CoNS isolated from cerebrospinal fluid cultures in 2012 to 2020 were recruited. One episode was defined as one patient with one bacterial strain. Interpretive categories were applied according to the MICs. The clinical outcomes were dichotomized into poor (Glasgow Outcome Scale 1 to 3) and acceptable (Glasgow Outcome Scale 4 to 5). In total, 534 episodes involving 519 patients and 16 bacteria were analyzed. Over the 9 years, eight antimicrobial agents were used in antimicrobial susceptibility tests, including six in over 80% of CoNS. The range of resistance rates was 0.8% to 84.6%. The vancomycin resistance rate was the lowest, whereas the penicillin resistance rate was the highest. The linezolid (a vancomycin replacement) resistance rate was 3.1%. The rate of oxacillin resistance, representing methicillin-resistant staphylococci, was 70.2%. There were no significant trends of antimicrobial susceptibility over the 9 years for any agents analyzed. However, there were some apparent changes. Notably, vancomycin-resistant CoNS appeared in recent years, while linezolid-resistant CoNS appeared early and disappeared in recent years. Vancomycin (or norvancomycin), the most common treatment agent, was used in 528 (98.9%) episodes. Finally, 527 (98.7%) episodes had acceptable outcomes. It will be safe to use vancomycin to treat CoNS-related HCAVM in the immediate future, although continuous monitoring will be needed. IMPORTANCE Coagulase-negative staphylococci are the main pathogens in health care-associated ventriculitis and meningitis. There are three conclusions from the results of this study. First, according to antimicrobial susceptibility, the rates of resistance to primary antimicrobial agents are high and those to high-level agents, including vancomycin, are low. Second, the trends of resistance rates are acceptable, especially for high-level agents, although long-term and continuous monitoring is necessary. Finally, the clinical outcomes of neurosurgical adults with coagulase-negative staphylococci-related health care-associated ventriculitis and meningitis are acceptable after treatment with vancomycin. Therefore, according to the antimicrobial susceptibility and clinical practice, vancomycin will be safe to treat coagulase-negative staphylococci-related health care-associated ventriculitis and meningitis.

Asunto(s)

Ventriculitis Cerebral/microbiología , Líquido Cefalorraquídeo/microbiología , Infección Hospitalaria/microbiología , Meningitis Bacterianas/microbiología , Infecciones Estafilocócicas/microbiología , Staphylococcus/efectos de los fármacos , Adolescente , Adulto , Anciano , Anciano de 80 o más Años , Antibacterianos/farmacología , Proteínas Bacterianas/genética , Proteínas Bacterianas/metabolismo , Coagulasa/genética , Coagulasa/metabolismo , Farmacorresistencia Bacteriana , Femenino , Humanos , Linezolid/farmacología , Masculino , Meningitis Bacterianas/líquido cefalorraquídeo , Pruebas de Sensibilidad Microbiana , Persona de Mediana Edad , Infecciones Estafilocócicas/líquido cefalorraquídeo , Staphylococcus/clasificación , Staphylococcus/genética , Staphylococcus/aislamiento & purificación , Vancomicina/farmacología , Adulto Joven

RESUMEN

A novel coagulase-negative Staphylococcus strain (NTUH-S172T) was isolated from human blood culture in Taiwan with preliminary identification of Staphylococcus saprophyticus. 16S rRNA gene analysis and multilocus sequence analysis (MLSA) showed that NTUH-S172T was most closely related to Staphylococcus haemolyticus. The average nucleotide identity and digital DNA-DNA hybridization values with the whole genome sequence were <95 % and<70 % when compared to the related species. Strain NTUH-S172T could be distinguished from S. haemolyticus by urease production and from Staphylococcus borealis by nitrate reduction. In addition, the matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS) spectrum of NTHU-S172T was significantly different from that of S. haemolyticus, which could be used in clinical identification. In conclusion, it is proposed that this isolate represents a novel species, named Staphylococcus taiwanensis sp. nov., with type strain NTUH-S172T (=BCRC 81315T=JCM 34726T).

Asunto(s)

Sangre/microbiología , Ácidos Grasos , Filogenia , Staphylococcus , Técnicas de Tipificación Bacteriana , Composición de Base , ADN Bacteriano/genética , Ácidos Grasos/química , Humanos , Hibridación de Ácido Nucleico , ARN Ribosómico 16S/genética , Análisis de Secuencia de ADN , Staphylococcus/clasificación , Staphylococcus/aislamiento & purificación , Taiwán

RESUMEN

The host range of bacteriophages defines their impact on bacterial communities and genome diversity. Here, we characterize 94 novel staphylococcal phages from wastewater and establish their host range on a diversified panel of 117 staphylococci from 29 species. Using this high-resolution phage-bacteria interaction matrix, we unveil a multi-species host range as a dominant trait of the isolated staphylococcal phages. Phage genome sequencing shows this pattern to prevail irrespective of taxonomy. Network analysis between phage-infected bacteria reveals that hosts from multiple species, ecosystems, and drug-resistance phenotypes share numerous phages. Lastly, we show that phages throughout this network can package foreign genetic material enclosing an antibiotic resistance marker at various frequencies. Our findings indicate a weak host specialism of the tested phages, and therefore their potential to promote horizontal gene transfer in this environment.

Asunto(s)

Genoma Viral , Especificidad del Huésped , Fagos de Staphylococcus/genética , Staphylococcus/genética , Aguas Residuales , Antibacterianos/farmacología , Farmacorresistencia Bacteriana Múltiple/genética , Ecosistema , Transferencia de Gen Horizontal , Consorcios Microbianos/genética , Pruebas de Sensibilidad Microbiana , Filogenia , Staphylococcus/clasificación , Staphylococcus/efectos de los fármacos , Staphylococcus/virología , Fagos de Staphylococcus/clasificación , Fagos de Staphylococcus/aislamiento & purificación , Aguas Residuales/microbiología , Aguas Residuales/virología , Microbiología del Agua

RESUMEN

Staphylococcus argenteus and Staphylococcus schweitzeri are the newest members of the Staphylococcus aureus complex. The number of clinical reports attributed to these new S. aureus complex members is limited. In a retrospective clinical laboratory study conducted over a 4-month period investigating the prevalence of S. argenteus and S. schweitzeri, a total of 43 isolates were selected. Phylogeny based on core-gene multilocus sequence typing (MLST) analysis confirmed that 37 were S. argenteus but a genetically distinct clade of six isolates was identified. Digital DNA-DNA hybridization (dDDH) and average nucleotide identity (ANI) analyses further supported the classification of these six isolates as a separate species. When compared to S. aureus complex reference genomes, the ANI values were ≤94 % and the dDDH values were <53 %. Based on the seven-gene S. aureus MLST scheme, the six isolates belong to five novel allelic profiles (ST6105, ST6106, ST6107, ST6108 and ST109). Their clinical infection features were similar to S. aureus. Skin and soft tissue infections presented in four out of the six cases. Routine clinical diagnostic identification using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry and biochemical profiling does not differentiate these new members from the rest of the complex. Genotypic analysis suggests that the six isolates belong to a novel species, Staphylococcus singaporensis sp. nov. with isolate SS21T (=DSM 111408T=NCTC14419T) designated as the type strain.

Asunto(s)

Filogenia , Staphylococcus/clasificación , Técnicas de Tipificación Bacteriana , Composición de Base , ADN Bacteriano/genética , Humanos , Tipificación de Secuencias Multilocus , Hibridación de Ácido Nucleico , ARN Ribosómico 16S/genética , Estudios Retrospectivos , Análisis de Secuencia de ADN , Staphylococcus/aislamiento & purificación , Staphylococcus aureus/genética

RESUMEN

The bacterial genus Staphylococcus comprises a large group of pathogenic and nonpathogenic species associated with an array of host species. Staphylococci are differentiated into coagulase-positive or coagulase-negative groups based on the capacity to promote clotting of plasma, a phenotype historically associated with the ability to cause disease. However, the genetic basis of this important diagnostic and pathogenic trait across the genus has not been examined to date. Here, we selected 54 representative staphylococcal species and subspecies to examine coagulation of plasma derived from six representative host species. In total, 13 staphylococcal species mediated coagulation of plasma from at least one host species including one previously identified as coagulase negative (Staphylococcus condimenti). Comparative genomic analysis revealed that coagulase activity correlated with the presence of a gene (vwb) encoding the von Willebrand binding protein (vWbp) whereas only the Staphylococcus aureus complex contained a gene encoding staphylocoagulase (Coa), the classical mediator of coagulation. Importantly, S. aureus retained vwb-dependent coagulase activity in an S. aureus strain deleted for coa whereas deletion of vwb in Staphylococcus pseudintermedius resulted in loss of coagulase activity. Whole-genome-based phylogenetic reconstruction of the Staphylococcus genus revealed that the vwb gene has been acquired on at least four different occasions during the evolution of the Staphylococcus genus followed by allelic diversification via mutation and recombination. Allelic variants of vWbp from selected coagulase-positive staphylococci mediated coagulation in a host-dependent manner indicative of host-adaptive evolution. Taken together, we have determined the genetic and evolutionary basis of staphylococcal coagulation, revealing vWbp to be its archetypal determinant. IMPORTANCE The ability of some species of staphylococci to promote coagulation of plasma is a key pathogenic and diagnostic trait. Here, we provide a comprehensive analysis of the coagulase positivity of the staphylococci and its evolutionary genetic basis. We demonstrate that the von Willebrand binding protein rather than staphylocoagulase is the archetypal coagulation factor of the staphylococci and that the vwb gene has been acquired several times independently during the evolution of the staphylococci. Subsequently, vwb has undergone adaptive diversification to facilitate host-specific functionality. Our findings provide important insights into the evolution of pathogenicity among the staphylococci and the genetic basis for a defining diagnostic phenotype.

Asunto(s)

Proteínas Bacterianas/genética , Coagulasa/genética , Coagulasa/metabolismo , Evolución Molecular , Staphylococcus/enzimología , Staphylococcus/genética , Animales , Aves , Coagulación Sanguínea , Genoma Bacteriano , Genómica/métodos , Caballos , Humanos , Filogenia , Conejos , Infecciones Estafilocócicas/microbiología , Infecciones Estafilocócicas/veterinaria , Staphylococcus/clasificación , Staphylococcus/metabolismo , Porcinos , Factores de Virulencia/genética