Plasmid-Mediated Quinolone Resistance in Different Diarrheagenic Escherichia coli Pathotypes Responsible for Complicated, Noncomplicated, and Traveler's Diarrhea Cases

This work was supported by grants MPY-1042/14 and PI14CIII/00051 from the Fondo de Investigaciones Sanitarias from the Spanish Ministry of Economy and Competitiveness. Sergio Sánchez acknowledges the Miguel Servet Programme of the Fondo de Investigaciones Sanitarias, Spanish Ministry of Economy and Competitiveness, for his research contract (CP13/00237).S

Salmonella enterica serovars Typhimurium and Enteritidis causing mixed infections in febrile children in Mozambique

Background and purpose: Invasive nontyphoidal salmonellosis, mostly caused by serovars Typhimurium and Enteritidis of Salmonella enterica, has emerged as a major public health problem in sub-Saharan Africa. The aim of this study was the clinical and microbiological characterization of nontyphoidal salmonellosis episodes affecting febrile children in Mozambique. Patients and methods: The clinical records of the patients were evaluated, and S. enterica isolates were characterized with regard to serovar, phage type, antimicrobial resistance (phenotype/responsible genes), plasmid content, pulsed-field gel electrophoresis, and multilocus sequence typing. Results: Fifteen S. Typhimurium and 21 S. Enteritidis isolates were recovered from blood samples of 25 children, the majority with underlying risk factors. With regard to phage typing, most isolates were either untypeable or reacted but did not conform, revealing that a number of previously unrecognized patterns are circulating in Mozambique. Most isolates were multidrug-resistant, with nearly all of the responsible genes located on derivatives of serovar-specific virulence plasmids. ST313 and ST11 were the predominant sequence types associated with S. Typhimurium and S. Enteritidis, respectively, and the uncommon ST1479 was also detected in S. Enteritidis. A distinct XbaI fragment of ~350 kb was associated with pulsed-field gel electrophoresis patterns of multidrug-resistant isolates of S. Enteritidis. Nearly half of the children were coinfected with both serovars, a fact expected to aggravate the disease and hamper the treatment. However, particularly poor outcomes were not observed for the coinfected patients. Conclusion: Mixed Salmonella infections could frequently occur in febrile children in Mozambique. Additional studies are required to determine their actual impact and consequences, not only in this country, but also in other African countries

Nationwide outbreak of Salmonella enterica serotype Kedougou infection in infants linked to infant formula milk, Spain, 2008

On 5 August 2008, the National Reference Laboratory of Salmonella (NRLS) noted an increase in the number of isolates of Salmonella enterica serotype Kedougou. As of 22 August, 29 isolates have been reported during 2008, which is ten times more than the average number of isolates identified by the NRLS during 2002-2007. All isolates have a typical, indistinguishable Pulse Field pattern (SALKEDXB-1, Spanish code) and are fully sensitive to the standard suite of antimicrobials.S

Epidemiological tracing of Salmonella enterica serotype Abortusovis from Spanish ovine flocks by PFGE fingerprinting

Salmonella enterica serotype Abortusovis, an ovine host-specific serotype, rare in most countries, is responsible for epidemic abortion episodes in Spain. With the aim of surveillance and detection of the spread of specific clones, 55 Abortusovis isolates collected during 1996-2001 from flocks in 11 provinces, were studied using XbaI-PFGE. Despite the fact that the strains were geographically and spatially related, PFGE demonstrated an epidemiologically acceptable discriminating power, identifying 20 clones (similarity, 52-96%). Clones Sabv6, 1, 5 were disseminated in seven, five and two areas respectively, while another 17 clones appeared in single places. Clones from nearby geographic regions showed a high relatedness (one band of difference in the PFGE profile) Sabv1-2-3, Sabv5-6, Sabv7-8, and Sabv13-14, suggesting a common ancestor. Co-isolation in the same flock (Sabv5-6, Sabv1-3, Sabv1-6) was detected. PFGE surveillance detected the predominance and widespread distribution of clone Sabv6 in 21 out of the 55 Abortusovis serotype episodes studied in Spain.We thank all the Spanish laboratories which sent Abortusovis strains to LNRSSE. This work was supported by a grant from the Instituto de Salud Carlos III (02/0008) and Junta de Andalucía (Investigation Group AGR-149).S

Cluster investigation of mixed O76:H19 Shiga toxin-producing Escherichia coli and atypical enteropathogenic E. coli infection in a Spanish household

A Spanish household was identified through a Public Health follow up on a Shiga toxin-producing Escherichia coli (STEC)-positive 14-month-old girl reporting bloody diarrhoea, with the four household members experiencing either symptomatic or asymptomatic STEC and/or atypical enteropathogenic E. coli (aEPEC) shedding. In total, two different O76:H19 STEC strains and six aEPEC strains belonging to multiple serotypes were isolated and characterized in the household during a 5-month period. Prolonged asymptomatic shedding of O76:H19 STEC and O51:H49 aEPEC was detected in two family members. Although there was no conclusive evidence, consumption of vegetables fertilized with sheep manure was the suspected source of infection. This study highlights the risk of cross-infections posed by prolonged asymptomatic carriage and close household contact between family members, and illustrates the importance of molecular epidemiology in understanding disease clusters.We thank José Manuel Luquin and Gemma Poignonfor facilitating the follow-up sampling of the house-hold members and relatives. We thank DanielEibach for critically reviewing the manuscript. Wealso thank Flemming Scheutz for conventional O:Hserotyping the strains. Sergio Sánchez acknowledgesthe Juan de la Cierva programme from theMinisterio de Economía y Competitividad for hisresearch contract. This study was supported by theMadrid Regional Government (P2009/AGR-1489)

Comparative whole-genome sequence analysis of Mycobacterium tuberculosis isolated from pulmonary tuberculosis and tuberculous lymphadenitis patients in Northwest Ethiopia

Background: Tuberculosis (TB), caused by the Mycobacterium tuberculosis complex (MTBC), is a chronic infectious disease with both pulmonary and extrapulmonary forms. This study set out to investigate and compare the genomic diversity and transmission dynamics of Mycobacterium tuberculosis (Mtb) isolates obtained from tuberculous lymphadenitis (TBLN) and pulmonary TB (PTB) cases in Northwest Ethiopia. Methods: A facility-based cross-sectional study was conducted using two groups of samples collected between February 2021 and June 2022 (Group 1) and between June 2020 and June 2022 (Group 2) in Northwest Ethiopia. Deoxyribonucleic acid (DNA) was extracted from 200 heat-inactivated Mtb isolates. Whole-genome sequencing (WGS) was performed from 161 isolates having ≥1 ng DNA/μl using Illumina NovaSeq 6000 technology. Results: From the total 161 isolates sequenced, 146 Mtb isolates were successfully genotyped into three lineages (L) and 18 sub-lineages. The Euro-American (EA, L4) lineage was the prevailing (n = 100; 68.5%) followed by Central Asian (CAS, L3, n = 43; 25.3%) and then L7 (n = 3; 2.05%). The L4.2.2.ETH sub-lineage accounted for 19.9%, while Haarlem estimated at 13.7%. The phylogenetic tree revealed distinct Mtb clusters between PTB and TBLN isolates even though there was no difference at lineages and sub-lineages levels. The clustering rate (CR) and recent transmission index (RTI) for PTB were 30 and 15%, respectively. Similarly, the CR and RTI for TBLN were 31.1 and 18 %, respectively. Conclusion and recommendations: PTB and TBLN isolates showed no Mtb lineages and sub-lineages difference. However, at the threshold of five allelic distances, Mtb isolates obtained from PTB and TBLN form distinct complexes in the phylogenetic tree, which indicates the presence of Mtb genomic variation among the two clinical forms. The high rate of clustering and RTI among TBLN implied that TBLN was likely the result of recent transmission and/or reactivation from short latency. Hence, the high incidence rate of TBLN in the Amhara region could be the result of Mtb genomic diversity and rapid clinical progression from primary infection and/or short latency. To validate this conclusion, a similar community-based study with a large sample size and better sampling technique is highly desirable. Additionally, analysis of genomic variants other than phylogenetic informative regions could give insightful information. Combined analysis of the host and the pathogen genome (GXG) together with environmental (GxGxE) factors could give comprehensive co-evolutionary information.The sample collection was funded by the Institute of Biotechnology, Bahir Dar University through the EN mega project. The Mtb culture and identification-related lab supply were supported by Amhara Public Health Institute, Bahir Dar Ethiopia. The whole-genome sequencing (WGS) and publication fee was covered by the National Center of Microbiology, Institute of Carlos III, Madrid, Spain. International Federation for Clinical Chemistry (IFCC) gave financial support to DM through the IFCC Professional Scientific Exchange Programme (PSEP) for 3-month WGS laboratory work.S

Un anticuerpo monoclonal específico frente al antígeno H:1,2 de Salmonella enterica serotipo Typhimurium que presenta reactividad cruzada con Salmonella enterica serotipo 4,5,12:i:-, línea celular productora, y su uso.

Esta invención se refiere a un anticuerpo monoclonal específico frente al antígeno H:1,2 presente en Salmonella entérica serotipo Typhimurium con reactividad cruzada frente al serotipo 4,5,12:i:-, una línea celular murina productora de dicho anticuerpo monoclonal y la aplicación de dicho anticuerpo monoclonal mediante técnicas inmunológicas para reconocer antígenos asociados a flagelo específicos de Salmonella enterica serotipo Typhimurium y Salmonella enterica serotipo 4,5,12:i:-.REIVINDICACIONES: 1. Un procedimiento para la obtención de un anticuerpo monoclonal, fragmento o variante del mismo caracterizado porque comprende las siguientes etapas: a) inmunizar un animal con el antígeno flagelar H:1, 2 aislado; b) inmortalizar los linfocitos B procedentes de los ratones inmunizados en el paso anterior, mediante su fusión con células de mieloma para dar lugar a un hibridoma; c) aislamiento de aquellos clones de hibridomas que secretan anticuerpos monoclonales reactivos frente al antígeno flagelar H:1, 2; d) cultivo de dichos clones celulares y posterior obtención del anticuerpo monoclonal de interés, fragmento o variante del mismo, caracterizado porque dicho anticuerpo monoclonal, fragmento o variante del mismo reacciona específicamente frente al antígeno flagelar H:1, 2 presente en Salmonella enterica serotipo Typhimurium y de forma cruzada con el serotipo 4, 5, 12:i:-. 2. Un anticuerpo monoclonal, fragmento o variante del mismo obtenido según el procedimiento de la reivindicación 1 caracterizado porque reacciona específicamente frente al antígeno flagelar H:1, 2 presente en Salmonella enterica serotipo Typhimurium y de forma cruzada con el serotipo 4, 5, 12:i:-. 3. Un anticuerpo monoclonal, fragmento o variante del mismo según la reivindicación 2, caracterizado porque el animal inmunizado para su obtención está seleccionado entre: conejo, cabra y ratón. 4. Un anticuerpo monoclonal, fragmento o variante del mismo según la reivindicación 2, caracterizado porque dicho anticuerpo posee un isotipo seleccionado entre: IgG, e IgM. 5. Un anticuerpo monoclonal, fragmento o variante del mismo según una de las reivindicaciones 2 a 4, caracterizado porque el hibridoma aislado en el paso c) de su procedimiento de obtención es la línea celular 23D4, ECACC no. 05122301. 6. Un anticuerpo monoclonal, fragmento o variante del mismo según la reivindicación 5, caracterizado porque dicho anticuerpo es murino. 7. Un anticuerpo monoclonal, fragmento o variante del mismo según una de las reivindicaciones 5 o 6, caracterizado porque dicho anticuerpo es del isotipo IgM. 8. Un anticuerpo monoclonal, fragmento o variante del mismo según una de las reivindicaciones 2 a 7, caracterizado porque dicho anticuerpo monoclonal se encuentra en sobrenadante, ascites o purificado. 9. Un fragmento de un anticuerpo monoclonal según una cualquiera de las reivindicaciones 2 a 8 caracterizado porque está seleccionado entre: un fragmento Fab, un fragmento F (ab') 2 y un fragmento Fv. 10. Una línea celular capaz de producir un anticuerpo monoclonal que reacciona específicamente frente al antígeno H:1, 2 presente en el serotipo Typhimurium y de forma cruzada con el serotipo 4, 5, 12:i:- de Salmonella enterica definido en las reivindicaciones 2 a 8 caracterizada porque dicha línea celular es la línea 23D4, ECACC no. 05122301. 11. Uso de un anticuerpo, fragmento o variante del mismo según una de las reivindicaciones 2 a 9, para detectar la presencia de los serotipos Typhimurium o 4, 5, 12:i:- de Salmonella enterica en una muestra. 12. Uso de un anticuerpo, fragmento o variante del mismo según la reivindicación 11, caracterizado porque dicha muestra está seleccionada entre una muestra alimentaria y una muestra clínica. 13. Uso de un anticuerpo, fragmento o variante del mismo según una de las reivindicaciones 11 u 12, caracterizado porque dicha muestra clínica está seleccionada entre sangre, suero, plasma y orina. 14. Uso de un anticuerpo monoclonal, un fragmento o variante del mismo según una de las reivindicaciones 11 a 13, caracterizado porque dicha detección se realiza mediante un ensayo seleccionado entre: ELISA, Western-blot y Dot-blot.Cuando una patente se hace internacional, se puede encontrar en el idioma de cada país en que se ha solicitado. En Espacenet se tiene acceso a los documentos en cada idioma.Universidad del País Vasco; Instituto de Salud Carlos IIISolicitud de patent

Zoonotic Transmission of Diphtheria from Domestic Animal Reservoir, Spain

Toxigenic Corynebacterium ulcerans is as an emerging zoonotic agent of diphtheria. We describe the zoonotic transmission of diphtheria caused by toxigenic C. ulcerans from domestic animals in Spain, confirmed by core-genome multilocus sequence typing. Alongside an increasing number of recent publications, our findings highlight the public health threat posed by diphtheria reemergence.This work was partially funded by Área de Ganadería de la Dirección General de Agricultura, Ganadería y Alimentación de la Comunidad de Madrid.S