Carbapenemase-Producing Enterobacteriaceae Recovered from a Spanish River Ecosystem

The increasing resistance to carbapenems is an alarming threat in the fight against multiresistant bacteria. The dissemination properties of antimicrobial resistance genes are supported by their detection in a diverse population of bacteria, including strains isolated from the environment. The objective of this study was to investigate the presence of carbapenemase-producing Enterobacteriaceae (CPE) collected from a river ecosystem in the Barcelona metropolitan area (Spain). Identification of β-lactamases and other resistance determinants was determined as was the antimicrobial susceptibility profile. Moreover, screening of virulence factors, plasmid addiction systems, plasmid partition systems and replicon typing was performed. The results identified 8 isolates belonging to different species (Escherichia coli, Enterobacter cloacae, Klebsiella pneumoniae, Klebsiella oxytoca, Raoultella ornithinolytica). The most prevalent enzyme was KPC-2 (n = 6), followed by VIM-1 (n = 2) and IMI-2 (n = 1), whereas no OXA-48-type was detected. In addition, one strain was positive for both KPC-2 and VIM-1 enzymes. All the carbapenemase-encoding plasmids carried at least one plasmid addiction or partition system, being vagCD and parAB the most frequently detected, respectively. E. coli and K. pneumoniae isolates carried a low number of virulence-associated factors and none of the detected clones has previously been identified in the clinical setting. These findings support the high dissemination potential of the carbapanemase-encoding genes and reinforce the idea that the environment is another reservoir that may play an important role in the capture, selection and dissemination of carbapenem resistance genes

Report of the Working Group on Acoustic and Egg Surveys for Sardine and Anchovy in ICES Areas 7, 8, and 9

The increasing resistance to carbapenems is an alarming threat in the fight against multiresistant bacteria. The dissemination properties of antimicrobial resistance genes are supported by their detection in a diverse population of bacteria, including strains isolated from the environment. The objective of this study was to investigate the presence of carbapenemase-producing Enterobacteriaceae (CPE) collected from a river ecosystem in the Barcelona metropolitan area (Spain). Identification of β-lactamases and other resistance determinants was determined as was the antimicrobial susceptibility profile. Moreover, screening of virulence factors, plasmid addiction systems, plasmid partition systems and replicon typing was performed. The results identified 8 isolates belonging to different species (Escherichia coli, Enterobacter cloacae, Klebsiella pneumoniae, Klebsiella oxytoca, Raoultella ornithinolytica). The most prevalent enzyme was KPC-2 (n = 6), followed by VIM-1 (n = 2) and IMI-2 (n = 1), whereas no OXA-48-type was detected. In addition, one strain was positive for both KPC-2 and VIM-1 enzymes. All the carbapenemase-encoding plasmids carried at least one plasmid addiction or partition system, being vagCD and parAB the most frequently detected, respectively. E. coli and K. pneumoniae isolates carried a low number of virulence-associated factors and none of the detected clones has previously been identified in the clinical setting. These findings support the high dissemination potential of the carbapanemase-encoding genes and reinforce the idea that the environment is another reservoir that may play an important role in the capture, selection and dissemination of carbapenem resistance genes

Increasing trend of antimicrobial resistance in Shigella associated with MSM transmission in Barcelona, 2020–21: outbreak of XRD Shigella sonnei and dissemination of ESBL-producing Shigella flexneri

Antimicrobial resistance; Shigella sonnei; Sexually transmitted infectionResistencia antimicrobiana; Shigella sonnei; Infección de transmisión sexualResistència antimicrobiana; Shigella sonnei; Infecció de transmissió sexualBackground Several countries have recently reported the detection of ESBL-producing Shigella sonnei associated with transmission among MSM. In a previous study by our group, 2.8% of Shigella spp. obtained from MSM in Barcelona between 2015 and 2019 were ESBL producers. Objectives To describe and characterize the emerging ESBL-producing Shigella spp. associated with sexual transmission among MSM detected from 2020 to 2021 in Barcelona, elucidating their connectivity with contemporaneous ESBL-producing Shigella spp. from other countries. Results From 2020 to 2021, we identified that among MSM, 68% of S. sonnei were XDR harbouring blaCTX-M-27 and 14% of Shigella flexneri were MDR harbouring blaCTX-M-27. WGS analysis showed that the ESBL-producing S. sonnei were part of a monophyletic cluster, which included isolates responsible for the prolonged outbreak occurring in the UK. Our data also reveal the first emergence and clonal dissemination of ESBL-producing and fluoroquinolone-resistant S. flexneri 2a among MSM. Conclusions We report an increasing trend of antimicrobial resistance in Shigella spp. among MSM in Barcelona since 2021, mainly as a consequence of the dissemination of XDR ESBL-producing S. sonnei, previously reported in the UK. These results highlight the importance of international collaborative surveillance of MDR/XDR S. sonnei and S. flexneri for rapid identification of their emergence and the prevention of the transmission of these pathogens.This work was partially supported by the ‘Ministerio de Economía y Competitividad’, ‘Instituto de Salud Carlos III’, and co-financed by the European Regional Development Fund (ERDF) ‘A Way to Achieve Europe’ (Spanish Network for Research in Infectious Diseases, grant number RD16/0016/0003) and by the Centro de Investigación Biomédica en Red (CIBER de Enfermedades Infecciosas, grant no. CB21/13/00054). A.M.M. is supported by a grant from the ‘Fondo de Investigación Sanitaria’ (Contratos Predoctorales de Formación en Investigación, grant number FI19/00315)

Increasing trend of antimicrobial resistance in Shigella associated with MSM transmission in Barcelona, 2020-21 : outbreak of XRD Shigella sonnei and dissemination of ESBL-producing Shigella flexneri

Several countries have recently reported the detection of ESBL-producing Shigella sonnei associated with transmission among MSM. In a previous study by our group, 2.8% of Shigella spp. obtained from MSM in Barcelona between 2015 and 2019 were ESBL producers. To describe and characterize the emerging ESBL-producing Shigella spp. associated with sexual transmission among MSM detected from 2020 to 2021 in Barcelona, elucidating their connectivity with contemporaneous ESBL-producing Shigella spp. from other countries. From 2020 to 2021, we identified that among MSM, 68% of S. sonnei were XDR harbouring bla and 14% of Shigella flexneri were MDR harbouring bla . WGS analysis showed that the ESBL-producing S. sonnei were part of a monophyletic cluster, which included isolates responsible for the prolonged outbreak occurring in the UK. Our data also reveal the first emergence and clonal dissemination of ESBL-producing and fluoroquinolone-resistant S. flexneri 2a among MSM. We report an increasing trend of antimicrobial resistance in Shigella spp. among MSM in Barcelona since 2021, mainly as a consequence of the dissemination of XDR ESBL-producing S. sonnei, previously reported in the UK. These results highlight the importance of international collaborative surveillance of MDR/XDR S. sonnei and S. flexneri for rapid identification of their emergence and the prevention of the transmission of these pathogens

Pertactin-Deficient Bordetella pertussis with Unusual Mechanism of Pertactin Disruption, Spain, 1986-2018

Bordetella pertussis not expressing pertactin has increased in countries using acellular pertussis vaccines (ACV). The deficiency is mostly caused by pertactin gene disruption by IS481. To assess the effect of the transition from whole-cell vaccine to ACV on the emergence of B. pertussis not expressing pertactin in Spain, we studied 342 isolates collected during 1986-2018. We identified 93 pertactin-deficient isolates. All were detected after introduction of ACV and represented 38% of isolates collected during the ACV period; 58.1% belonged to a genetic cluster of isolates carrying the unusual prn::del(-292, 1340) mutation. Pertactin inactivation by IS481 insertion was identified in 23.7% of pertactin-deficient isolates, arising independently multiple times and in different phylogenetic branches. Our findings support the emergence and dissemination of a cluster of B. pertussis with an infrequent mechanism of pertactin disruption in Spain, probably resulting from introduction of ACV.This work was supported by the Ministerio de Economía y Competitividad, Instituto de Salud Carlos III, and cofinanced by the European Regional Development Fund “A Way to Achieve Europe” (Spanish Network for Research in Infectious Diseases, grant no. FIS PI18/00703) and by the Centro de Investigación Biomédica en Red (CIBER de Enfermedades Infecciosas), the Red Española de Investigación en Patología Infecciosa (grant no. CB21/13/00054). A.M.C. is supported by the Agència de Gestió d’Ajuts Universitaris i de Recerca de la Generalitat de Catalunya at Vall d’Hebron Institut de Recerca (Ajuts per a la Contractació de Personal Investigador FI, grant no. 2020FI_B2_00145) and by the Spanish Network for Research in Infectious Diseases (grant no. RD16/ 0016/0003). A.M.M. is supported by a grant from the Fondo de Investigación Sanitaria at Vall d’Hebron Institut de Recerca (Contratos Predoctorales de Formación en Investigación, grant no. FI19/00315).S

Dinàmica poblacional i deriva antigènica de Bordetella pertussis i el paper d'altres espècies del gènere Bordetella en l'emergència de la tosferina al segle XXI

La tosferina és una malaltia infecciosa aguda causada generalment per Bordetella pertussis, un cocbacil gramnegatiu, de reservori exclusivament humà i amb una elevada transmissibilitat per via aèria. Es caracteritza per presentar un quadre respiratori de tos intensa que, sovint, pot perllongar-se durant setmanes o mesos. Entre les possibles causes implicades en el ressorgiment de la tosferina, destaca l'adaptació de B. pertussis a la immunitat conferida per les vacunes acel·lulars (DTPa), les quals es van introduir en el nostre entorn a finals dels anys noranta, així com l'emergència de noves espècies del gènere Bordetella contra les quals les vacunes utilitzades no confereixen protecció. Per una banda, el procés d'adaptació de B. pertussis a la immunitat induïda per les vacunes DTPa ha evidenciat com, coincidint amb la introducció d'aquestes, han aparegut noves poblacions de B. pertussis que han anat reemplaçant de forma progressiva les variants antigèniques presents a les composicions vacunals utilitzades. En aquest sentit, s'ha observat un viratge en l'estructura de les poblacions de B. pertussis circulants, reflectida pel recanvi dels perfils electroforètics que han circulat entre els períodes d'ús de les vacunes de cèl·lules completes (DTPw) i les vacunes DTPa, així com de l'emergència i posterior predomini del genotip MT27, el qual s'ha observat en el 79,3% dels aïllats de B. pertussis circulants després de la introducció de les vacunes DTPa. Addicionalment, en aquestes poblacions, s'han observat polimorfismes en els gens que codifiquen la toxina pertussis (ptxA) i en el seu promotor (ptxP), la pertactina (prn) i la fimbria de tipus 3 (fim3). Així, la combinació al·lèlica ptxA1/prn2/fim3-2, la qual presenta formes antigèniques diferents a les que es troben contingudes en les vacunes DTPa, s'ha observat com a majoritària (52,7%) entre els aïllats que han circulat en el nostre medi després de la introducció d'aquest tipus de vacuna. Addicionalment, la presència majoritària (97,7%) del promotor de la toxina pertussis de tipus 3 (ptxP3) condiciona un increment de la producció d'aquest determinant de virulència en aquests aïllats de B. pertussis que el posseeixen. D'altra banda, s'ha posat de manifest la circulació d'aïllats de B. pertussis que deixen d'expressar l'antigen vacunal de la pertactina (PRN), mantenint la seva virulència i capacitat per produir malaltia. Aquests han suposat 27,2% del total d'aïllats, identificant-se únicament en el període d'ús de les vacunes DTPa. En aquest sentit, l'aparició i disseminació exitosa, d'un clúster d'aïllats que han presentat una deleció en el promotor i part del gen que codifica aquest determinant de virulència (58,1%, prn::del(-292, 1340)), ha evidenciat l'expansió dels aïllats de B. pertussis que no expressen PRN en el nostre medi. D'aquesta manera, la disseminació d'aïllats de B. pertussis que expressen antígens vacunals que posseeixen epítops no reconeguts pels anticossos generats mitjançant l'ús de les vacunes DTPa, així com la pèrdua de l'expressió d'aquests antígens com a importants mecanismes d'adaptació i evasió de la immunitat conferida per les vacunes antipertússiques utilitzades al llarg dels últims anys, sembla ser un mecanisme que pot comprometre la seva eficàcia. Finalment, la descripció de l'emergència de B. holmesii com a agent causal de la tosferina, observant-se una prevalença d'aquest microorganisme del 3,9% l'any 2015 i del 8,8% l'any 2016, demostra que la seva circulació ha pogut contribuir en el ressorgiment de la tosferina en el nostre medi. En relació amb aquests, no s'han observat diferències en les característiques clíniques-epidemiològiques dels casos de B. holmesii en comparació amb aquells causats per B. pertussis, ni s'han evidenciat complicacions o recidives després del seu tractament amb azitromicina. Així, el conjunt d'observacions reforça la necessitat d'adaptar les eines diagnòstiques actuals per la identificació correcta de les etiologies de la tosferina.La tosferina es una enfermedad infecciosa aguda causada generalmente por Bordetella pertussis, un cocobacilo gramnegativo, de reservorio exclusivamente humano y con una elevada transmisibilidad por vía aérea. Se caracteriza por presentar un cuadro respiratorio de tos intensa que, a menudo, puede prolongarse durante semanas o meses. Entre las posibles causas implicadas en el resurgimiento de la tosferina, destaca la adaptación de B. pertussis a la inmunidad conferida por las vacunas acelulares (DTPa), las cuales se introdujeron a finales de los años noventa, así como la emergencia de nuevas especies del género Bordetella contra las que las vacunas utilizadas no confieren protección. Por un lado, el proceso de adaptación de B. pertussis a la inmunidad inducida por las DTPa ha evidenciado como, coincidiendo con la introducción de éstas, han aparecido nuevas poblaciones de B. pertussis que han ido reemplazando progresivamente las variantes antigénicas presentes en las composiciones vacunales utilizadas. En este sentido, se ha observado un viraje en la estructura de las poblaciones de B. pertussis circulantes, reflejada por el recambio de los perfiles electroforéticos que han circulado entre los períodos de uso de las vacunas de células completas (DTPw) y las DTPa, así como de la emergencia y posterior predominio del genotipo MT27, el que se ha observado en el 79,3% de los aislados de B. pertussis circulantes tras la introducción de las DTPa. Adicionalmente, en estas poblaciones, se han observado polimorfismos en los genes que codifican la toxina pertusis (ptxA) y en su promotor (ptxP), la pertactina (prn) y la fimbria de tipo 3 (fim3). Así, la combinación alélica ptxA1/prn2/fim3-2, la cual presenta formas antigénicas diferentes a las contenidas en las DTPa, se ha observado como mayoritaria (52,7%) entre los aislados que han circulado en nuestro medio después de la introducción de estas vacunas. Adicionalmente, la presencia mayoritaria (97,7%) del promotor de la toxina pertusis de tipo 3 (ptxP3) condiciona un incremento de la producción de este determinante de virulencia entre los aislados de B. pertussis que lo poseen. Por otra parte, se ha puesto de manifiesto la circulación de aislados de B. pertussis que dejan de expresar el antígeno vacunal de la pertactina (PRN), manteniendo su virulencia y capacidad para producir enfermedad. Éstos han supuesto 27,2% del total, identificándose únicamente en el período de uso de las DTPa. En este sentido, la aparición y diseminación exitosa, de un clúster de aislados que han presentado una deleción en el promotor y parte del gen que codifica este determinante de virulencia (58,1%, prn::del(-292, 1340)), ha evidenciado la expansión de los aislados de B. pertussis que no expresan PRN. De este modo, la diseminación de aislados de B. pertussis que expresan antígenos vacunales que poseen epítopos no reconocidos por los anticuerpos generados mediante el uso de las DTPa, así como la pérdida de la expresión de estos antígenos como importantes mecanismos de adaptación y evasión de la inmunidad conferida por las vacunas utilizadas, parece ser un mecanismo que puede comprometer su eficacia. Finalmente, la descripción de la emergencia de B. holmesii como agente causal de la tosferina, observándose una prevalencia de este microorganismo del 3,9% en 2015 y del 8,8% en 2016, demuestra que su circulación ha podido contribuir en el resurgimiento de la tosferina. En relación con éstos, no se han observado diferencias en las características clínicas-epidemiológicas de los casos de B. holmesii en comparación con aquellos causados por B. pertussis, ni se han evidenciado complicaciones o recidivas después de su tratamiento con azitromicina. Así, el conjunto de observaciones refuerza la necesidad de adaptar las herramientas diagnósticas actuales para la identificación correcta de las etiologías de la tosferina.Whooping cough is an acute infectious disease usually caused by Bordetella pertussis, a gram-negative coccobacillus, with an exclusively human reservoir and high airborne transmissibility. It is characterized by presenting a respiratory picture of intense cough that can often last for weeks or months. Among the possible causes that are involved in the pertussis resurgence, the adaptation of B. pertussis to the immunity conferred by acellular vaccines (DTPa), which were introduced in the late 1990s, as well as the emergence of new species of the genus Bordetella, against which the vaccines used do not confer protection, are some of the most important ones that have be considered. On the one hand, the process of adaptation of B. pertussis to the immunity induced by DTPa has been evidenced by the new populations of B. pertussis that have appeared and have gradually replaced the antigenic variants present in the vaccine, concurrently with its introduction. In this sense, a change in the structure of circulating populations has been observed, reflected by the replacement of the electrophoretic profiles that have circulated within the different vaccine periods. Is remarkable the emergence and subsequent predominance of the MT27 genotype, which has been observed in 79.3% of circulating B. pertussis isolates after the introduction of DTPa. Additionally, in these populations, polymorphisms in the genes encoding the pertussis toxin (ptxA) and its promoter (ptxP), pertactin (prn) and fimbria type 3 (fim3). Thus, the allelic combination ptxA1/prn2/fim3-2, which include different antigenic forms than those contained in DTPa, has been observed as the most prevalent (52.7%) among the isolates that have circulated in our environment after the introduction of this type of vaccine. Additionally, a high prevalence (97.7%) of the type 3 pertussis toxin promoter (ptxP3), which induces an increase in the production of this virulence determinant in these B. pertussis isolates, is found in isolates obtained after the DTPa introduction. Other factor that has evidenced the B. pertussis adaptation is the emergence of pertactin (PRN) negative isolates maintaining its virulence and ability to produce disease. In our study, PRN-negative isolates have accounted for the 27.2% of the total studied isolates. They have been identified only during the period of use of DTPa. In this sense, the emergence and successful dissemination of a cluster of isolates that possess a deletion in the promoter and part of the gene encoding this determinant of virulence (58.1%, prn::del(-292 , 1340)), has evidenced the expansion of most of the B. pertussis isolates that do not express PRN. Thus, the spread of B. pertussis isolates expressing vaccine antigens possessing epitopes not recognized by antibodies generated by the use of DTPa, as well as the loss of expression of these antigens as an important mechanism of adaptation and evasion of immunity conferred by pertussis vaccines, seems to be a mechanism contributing to compromise the vaccine effectiveness. Finally, the description of the emergence of B. holmesii as the causative agent of pertussis, observing a prevalence of this microorganism of 3.9% in 2015 and 8.8% in 2016, shows that its circulation has been able to contribute to the pertussis resurgence. In relation to these, no differences were observed in the clinical-epidemiological characteristics of the cases by B. holmesii and those caused by B. pertussis, nor were there any complications or relapses after its treatment with azithromycin. Accurate diagnosis of the causative agent is crucial to determine the incidence and prevalence of the microbial species involved, to assess its contribution to the epidemiology of whooping cough, to evaluate whether specific antimicrobial drug treatments should be implemented and, in terms of public health, to assess the efficacy of the pertussis vaccine

Carbapenemase-Producing Enterobacteriaceae Recovered from a Spanish River Ecosystem

The increasing resistance to carbapenems is an alarming threat in the fight against multiresistant bacteria. The dissemination properties of antimicrobial resistance genes are supported by their detection in a diverse population of bacteria, including strains isolated from the environment. The objective of this study was to investigate the presence of carbapenemase-producing Enterobacteriaceae (CPE) collected from a river ecosystem in the Barcelona metropolitan area (Spain). Identification of β-lactamases and other resistance determinants was determined as was the antimicrobial susceptibility profile. Moreover, screening of virulence factors, plasmid addiction systems, plasmid partition systems and replicon typing was performed. The results identified 8 isolates belonging to different species (Escherichia coli, Enterobacter cloacae, Klebsiella pneumoniae, Klebsiella oxytoca, Raoultella ornithinolytica). The most prevalent enzyme was KPC-2 (n = 6), followed by VIM-1 (n = 2) and IMI-2 (n = 1), whereas no OXA-48-type was detected. In addition, one strain was positive for both KPC-2 and VIM-1 enzymes. All the carbapenemase-encoding plasmids carried at least one plasmid addiction or partition system, being vagCD and parAB the most frequently detected, respectively. E. coli and K. pneumoniae isolates carried a low number of virulence-associated factors and none of the detected clones has previously been identified in the clinical setting. These findings support the high dissemination potential of the carbapanemase-encoding genes and reinforce the idea that the environment is another reservoir that may play an important role in the capture, selection and dissemination of carbapenem resistance genes