Within-host diversity of coagulase-negative staphylococci resistome from healthy pigs and pig farmers, with the detection of cfr-carrying strains and MDR-S. borealis

The ecology and diversity of resistome in coagulase-negative staphylococci (CoNS) from healthy pigs and pig farmers are rarely available as most studies focused on the livestock-associated methicillin-resistant S. aureus. This study aims to characterize the antimicrobial resistance (AMR) mechanisms, intra-host species diversity (more than one species in a host), and intra-species AMR diversity (same species with more than one AMR profile) in CoNS recovered from the nasal cavities of healthy pigs and pig farmers. One-hundred-and-one CoNS strains previously recovered from 40 pigs and 10 pig farmers from four Spanish pig farms were tested to determine their AMR profiles. Non-repetitive strains were selected (n = 75) and their AMR genes, SCCmec types, and genetic lineages were analyzed by PCR/sequencing. Of the non-repetitive strains, 92% showed a multidrug resistance (MDR) phenotype, and 52% were mecA-positive, which were associated with SCCmec types V (46.2%), IVb (20.5%), and IVc (5.1%). A total of 28% of the pigs and pig farmers had intra-host species diversity, while 26% had intra-species AMR diversity. High repertoires of AMR genes were detected, including unusual ones such as tetO, ermT, erm43, and cfr. Most important was the detection of cfr (in S. saprophyticus and S. epidermidis-ST16) in pigs and pig farmers; whereas MDR-S. borealis strains were identified in pig farmers. Pig-to-pig transmission of CoNS with similar AMR genes and SCCmec types was detected in 42.5% of pigs. The high level of multidrug, within-host, and intra-species resistome diversity in the nasal CoNS highlights their ability to be AMR gene reservoirs in healthy pigs and pig farmers. The detection of MDR-S. borealis and linezolid-resistant strains underscore the need for comprehensive and continuous surveillance of MDR-CoNS at the pig farm level

Simultaneous Nasal Carriage by Methicillin-Resistant and Methicillin Susceptible Staphylococcus aureus of Lineage ST398 in a Live Pig Transporter.

Methicillin-resistant Staphylococcus aureus (MRSA) sequence type (ST)398 is a livestock associated (LA) lineage with zoonotic potential, especially in humans with live pig contact. The objective of this study was to characterize two S. aureus strains of lineage ST398 (one methicillin-resistant (MRSA), one methicillin-susceptible (MSSA)) isolated from the same nasal sample of a patient admitted in the Intensive-Care Unit of a Spanish Hospital, and with previous occupational exposure to live pigs, by whole-genome-sequencing (WGS). The sample was obtained during routine surveillance for MRSA colonization. Purified genomic DNA was sequenced using Illumina HiSeq 2000 and processed using conventional bioinformatics software. The two isolates recovered were both S. aureus t011/ST398 and showed similar resistance-phenotypes, other than methicillin susceptibility. The possession of antibiotic resistance genes was the same, except for the mecA-gene located in SCCmecV in the MRSA isolate. The MSSA isolate harbored remnants of a SCCmec following the deletion of 17342bp from a recombination between two putative primases. Both isolates belonged to the livestock-associated clade as defined by three canonical single-nucleotide-polymorphisms, and neither possessed the human immune evasion cluster genes, chp, scn, or sak. The core genome alignment showed a similarity of 99.6%, and both isolates harbored the same mobile genetic elements. The two nasal ST398 isolates recovered from the patient with previous occupational exposure to pigs appeared to have a livestock origin and could represent different evolutionary steps of animal-human interface lineage. The MSSA strain was formed as a result of the loss of the mecA gene from the livestock-associated-MRSA lineage

Soluble proteins and free amino nitrogen content in must and wine of cv. Viura in La Rioja

The protein fraction of white musts and wines obtained from grapes (Vitis vinifera L. cv. Viura) grown in northern Spain (Rioja) was investigated by SDS-PAGE. Under different fermentation conditions, e.g. aeration, must nitrogen content and yeast strain, SDS-PAGE patterns showed several bands with molecular weights ranging from 16 to 200 kDa. Higher weight proteins were glycosylated, whereas lower weight proteins were not. Under the experimental fermentation conditions none of the proteins showed any chemical modification that would alter the electrophoretical mobility or the covalent binding to their glycosylated moiety

Detección y caracterización de cepas Escherichia coli enteropatógena (EPEC) aisladas de la cavidad nasal de cerdos potencialmente sanos

Introducción: Escherichia coli enteropatógena (EPEC) es un importante patógeno, implicado enprocesos diarreicos asociados a infecciones extraintestinales tanto en clínica humana como enveterinaria. Las cepas EPEC poseen el gen eae (codificante de intimina) y carecen de stx (toxinashiga), y pueden ser EPEC típicas/atípicas en función de la presencia/ausencia del gen bfp. Elobjetivo de este estudio fue analizar la frecuencia de E. coli y EPEC en muestras nasales de cerdospotencialmente sanos, así como la relación genética y los perfiles de sensibilidad a antibióticos delos aislados EPEC. Materiales y métodos: Se analizaron muestras nasales de 40 cerdos de cuatrogranjas de Aragón (10 cerdos/granja) y fueron procesadas para el estudio de la microbiota nasalmediante el cultivo de bacterias Gram positivas y negativas. Se aislaron cepas de E. coli en agarsangre y se determinó a) la sensibilidad a antibióticos (CLSI, 2022); b) presencia del gen eae y bfppor PCR y el genotipo de resistencia a antibióticos de los aislados EPEC. Resultados: Se detectóla presencia de E. coli en 17/40 cerdos analizados (42.5%) y 25 aislados fueron recuperados. Lapresencia de EPEC se reflejó en 7/40 cerdos (17.5%) de 2/4 granjas, con 10 aislados EPEC. Los 10aislados eae-positivos fueron atípicos y presentaron un fenotipo de multi-resistencia (incluyendoresistencia a ampicilina, cloranfenicol, gentamicina, tetraciclina, y trimetoprim/sulfametoxazol) conlos genes blaTEM/tet(B)/floR/cmlA/sul1/sul2/sul3. Diversos linajes genéticos fueron detectados entrelos aislados eae-positivos. Conclusión: En la cavidad nasal de los cerdos se detecta con frecuenciaE. coli portador de genes de resistencia y virulencia de alto impacto clínico, las cuales pueden sertransferidas al hombre por la cadena alimentaria o por contacto con los animales. Aunque E. colies una bacteria comensal del tracto gastrointestinal, el estilo de vida de los animales favorece latransferencia al entono nasal

Detección de Escherichia coli enteropatogénica (EPEC) en el tracto gastrointestinal de pollos de cigüeñas blancas (Ciconia ciconia)

La Organización Mundial de la Salud ha estimado que alrededor de dos millones de niños muerenanualmente de enfermedades diarreicas en países en vías de desarrollo. Entre los agentes etiológicos clásicos queproducen un cuadro diarreico importante se encuentra Escherichia coli enteropatogénica (EPEC). Se conoce que elprincipal reservorio natural es el ser humano, especialmente niños con diarrea y niños o adultos que son portadoresasintomáticos, pero también se han aislado EPEC de diversos serotipos de animales salvajes. Debido a la reconocidaimportancia que tiene EPEC a nivel epidemiológico, en este estudio se utilizan polluelos de cigüeña para evaluar cómoinfluyen diferentes dietas (basadas en vertederos o en zonas naturales) en el papel potencial de las cigüeñas blancas comoreservorios de EPE