Search

Vanesa García,1 Inácio Mandomando,2,3 Joaquim Ruiz,4 Silvia Herrera-León,5 Pedro L Alonso,3,4 M Rosario Rodicio1 1Departamento de Biología Funcional, Área de Microbiología, Universidad de Oviedo, Oviedo, Spain; 2Centro de Investigação em Saúde de Manhiça, 3Instituto Nacional de Saúde, Ministério da Saúde, Maputo, Mozambique; 4ISGlobal, Barcelona Centre for International Health Research, Hospital Clínic, Universitat de Barcelona, Barcelona, 5Centro Nacional de Microbiología, Instituto de Salud Carlos III, Majadahonda, Madrid, Spain 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. Keywords: invasive nontyphoidal salmonellosis, bloodstream infection, multidrug resistance, virulence-resistance plasmid, ST313, ST1479

Daniel Mekonnen,1,2 Abaineh Munshea,2,3 Endalkachew Nibret,2,3 Bethlehem Adnew,4 Hailu Getachew,5 Amiro Kebede,5 Ananya Gebrewahid,5 Silvia Herrera-Leon,6 Aranzazu Amor Aramendia,7 Agustín Benito,8 Estefanía Abascal,6 Camille Jacqueline,6,9 Abraham Aseffa,4 Laura Herrera-Leon6,10 1Department of Medical Laboratory Sciences, School of Health Science, College of Medicine and Health Sciences, Bahir Dar University, Bahir Dar, Ethiopia; 2Health Biotechnology Division, Institute of Biotechnology, Bahir Dar University, Bahir Dar, Ethiopia; 3Department of Biology, Bahir Dar University, Bahir Dar, Ethiopia; 4Armauer Hansen Research Institute, Addis Ababa, Ethiopia; 5Amhara Public Health Institute, Bahir Dar, Ethiopia; 6National Centre for Microbiology, Instituto de Salud Carlos III, Madrid, Spain; 7Fundación Mundo Sano, Madrid, Spain; 8National Center of Tropical Medicine, Institute of Health Carlos III, Centro de Investigación Biomédica En Red de Enfermedades Infecciosas, Madrid, Spain; 9European Public Health Microbiology Training Programme, European Centre for Disease Prevention and Control, Stockholm, Sweden; 10CIBER Epidemiologia y Salud Publica, Madrid, SpainCorrespondence: Daniel Mekonnen, Email nigusdaniel@gmail.comIntroduction: Drug resistance (DR) in Mycobacterium tuberculosis complex (MTBC) is mainly associated with certain lineages and varies across regions and countries. The Beijing genotype is the leading resistant lineage in Asia and western countries. M. tuberculosis (Mtb) (sub) lineages responsible for most drug resistance in Ethiopia are not well described. Hence, this study aimed to identify the leading drug resistance sub-lineages and characterize first-line anti-tuberculosis drug resistance-associated single nucleotide polymorphisms (SNPs).Methods: A facility-based cross-sectional study was conducted in 2020– 2022 among new and presumptive multidrug resistant-TB (MDR-TB) cases in Northwest Ethiopia. Whole-genome sequencing (WGS) was performed on 161 isolates using Illumina NovaSeq 6000 technology. The SNP mutations associated with drug resistance were identified using MtbSeq and TB profiler Bioinformatics softwares.Results: Of the 146 Mtb isolates that were successfully genotyped, 20 (13.7%) harbored one or more resistance-associated SNPs. L4.2.2.ETH was the leading drug-resistant sub-lineage, accounting for 10/20 (50%) of the resistant Mtb. MDR-TB isolates showed extensive mutations against first-line anti-TB drugs. Ser450Leu/(tcg/tTg) for Rifampicin (RIF), Ser315Thr/(agc/aCc) for Isoniazid (INH), Met306Ile/(atg/atA(C)) for Ethambutol (EMB), and Gly69Asp for Streptomycin (STR) were the leading resistance associated mutations which accounted for 56.5%, 89.5%, 47%, and 29.4%, respectively. The presence of both clustered and non-clustered drug resistance (DR) isolates indicated that the epidemics is driven by both new DR development and acquired resistance.Conclusion: The high prevalence of drug-resistant TB due to geographically restricted sub-lineages (L4.2.2.ETH) indicates the ongoing local micro epidemics. The Mtb drug resistance surveillance system must be improved. Further evolutionary analysis of L4.2.2.ETH strain is highly desirable to understand evolutionary forces that leads L4.2.2.ETH in to high level DR and transmissible sub-lineage.Keywords: Mycobacterium tuberculosis, L.4.2.2, SIT149, drug resistance, Ethiopia

The aim of this study was to investigate the resistance mechanisms of quinolones, macrolides and tetracycline in campylobacter isolates from grandparent and parent broiler breeders in Spain. Twenty-six isolates were investigated for quinolone resistance, three isolates for macrolide resistance and 39 for tetracycline resistance. All of the quinolone-resistant isolates possessed the mutation Thr86Ile in the quinolone resistance-determining region ofgyrA and one isolate possessed the mutation Pro104Ser. Only oneCampylobacter colipopulation (defined by restriction fragment length polymorphism–polymerase chain reaction offlaA and pulsed field gel electrophoresis) was resistant to erythromycin, and the mutation A2075G (23S rDNA) was responsible for macrolide resistance. ThetetO gene was found in all of the tetracycline-resistant isolates. Twenty-two out of the 39 isolates investigated by Southern blot possessed chromosomic location oftetO and 17 were located on plasmids. Most of the plasmids withtetO were of around 60 kb and conjugation was demonstrated in a selection of them. In conclusion, we showed that Thr86Ile is highly prevalent in quinolone-resistant isolates as well as mutation A2075G in macrolide-resistant isolates of poultry origin. More variability was found fortetO. The possibility of horizontal transmission oftetO among campylobacter isolates is also an issue of concern in public health. [ABSTRACT FROM AUTHOR]

The aim of this study was to determine the presence of virulence genes and antibiotic resistance profiles in 164Escherichia colistrains isolated from birds (feral pigeons, hybrid ducks, house sparrows and spotless starlings) inhabiting urban and rural environments. A total of eight atypical enteropathogenicE. colistrains were identified: one in a house sparrow, four in feral pigeons and three in spotless starlings. Antibiotic resistance was present in 32.9% (54) ofE. colistrains. The dominant type of resistance was to tetracycline (21.3%), ampicillin (19.5%) and sulfamethoxazole (18.9%). Five isolates had class 1 integrons containing gene cassettes encoding for dihydrofolate reductase A (dfrA) and aminoglycoside adenyltransferase A (aadA), one in a feral pigeon and four in spotless starlings. To our knowledge, the present study constitutes the first detection of virulence genes fromE. coliin spotless starlings and house sparrows, and is also the first identification worldwide of integrons containing antibiotic resistance gene cassettes inE. colistrains from spotless starlings and pigeons. [ABSTRACT FROM AUTHOR]

The epidemiology of subclinical salmonellosis in wild birds in a region of high Salmonella prevalence in pigs was studied. Three hundred and seventy-nine faecal samples from 921 birds trapped in 31 locations nearby pig premises, and 431 samples from 581 birds of 10 natural settings far from pig farms were analysed for the presence of Salmonella spp. Positive samples were serotyped and analysed for antimicrobial resistance (AR). Phage typing and pulsed-field gel electrophoresis (PFGE) on Salmonella Typhimurium isolates were also carried out. The overall proportion of Salmonella-positive samples was 1.85% (95% CI = 0.93, 2.77). Salmonella isolation was positively associated with samples collected from birds in the proximity of a pig operation (OR = 16.5; 95% CI = 5.17, 52.65), and from non-migratory (or short-distance migration) birds (OR = 7.6; 95% CI = 1.20, 48.04) and negatively related to mostly granivorous birds (OR = 0.4; 95% CI = 0.15, 1.13). Salmonella Typhimurium was the most prevalent serotype and four different XbaI PFGE patterns were observed that matched the four phage types identified (U310, U311, DT164 and DT56). Only 20% of the strains showed multi-AR. In three farms, a high degree of homogeneity among isolates from different birds was observed. These findings suggested that pig farms may act as amplifiers of this infection among wild birds, and the degree of bird density may have much to do on this transmission. Some of the Salmonella serotypes isolated from bird faeces were of potential zoonotic transmission and associated with AR. Monitoring salmonellosis in wild bird is advised. [ABSTRACT FROM AUTHOR]

The aims of this study were to ascertain the population structure and antimicrobial susceptibility of Salmonella enterica serovars isolated in 2002 from food in 16 Spanish regions. Serovars were characterized by serotyping, phage typing, antimicrobial susceptibility, and pulsed-field gel electrophoresis (PFGE) typing, and 264 nonrelated strains were selected for further analysis. The main sources were eggs and their derivatives (21.6% of strains), poultry and related products (16.6%), and seafood (16.3%). High serotype diversity was detected (51 serotypes); the most common were Enteritidis (n = 96, 36.3%) and Typhimurium (n = 53, 20.1%), followed by a miscellaneous group of 49 different serotypes (n = 115, 43.5%). A 15% increase in Salmonella Enteritidis isolation was observed. Common phage types for Salmonella Enteritidis were PTI (41.6% of isolates), PT4 (9.4%), PT6 (9.4%), and PT6a (9.4%), and common types for Salmonella Typhimurium were DTU302 (18.8%), DT104 (15.1%), and DT104B (13.2%). Salmonella Enteritidis strains were categorized into eight PFGE types with a similarity of 81 to 96%, and 73.9% of the strains were grouped into just one cluster. Salmonella Typhimurium isolates were divided into 13 PFGE types with a similarity of 64 to 86%, and one predominant clone contained 41.5% of the strains. Resistance rates for Salmonella Enteritidis, Salmonella Typhimurium, and the miscellaneous group were, respectively, 8.3, 69.8, and 13.9% for ampicillin, 3.1, 52.8, and 59% for streptomycin, 40.6, 22.6, and 10.4% for nalidixic acid, 15.6, 71.7, and 31.1% for tetracycline, 7.3, 18.8, and 9.5% for trimethoprim-sulfamethoxazole, 0, 50.9, and 4.3% for chloramphenicol, and 6.2, 71.7, and 17.4% for multiple (at least four) antimicrobials. Ail the strains remained susceptible to other β-lactams and fluoroquinolones. Surveillance of S. enterica isolated from food is strongly recommended to reduce community exposure to antimicrobial resistant strains. [ABSTRACT FROM AUTHOR]

Along with the increasing demand for lithium-ion batteries (LIB), the need for recycling major components such as graphite and different critical materials contained in LIB is also reaching a peak in the research community. Several authors review the different LIB recycling methodologies, including pyro- and hydrometallurgy processes. However, the characteristics, main stages, and achievements of LIB emerging recycling are still missing. This study reviews the diverse emerging approaches for recycling critical materials from spent LIB in the last five years. A classification for emerging recycling technologies is provided, including terms like development stage and eco-friendly status. The main stages of recycling LIB are opening, phase separation, and materials recovery. Among the emerging proposals with the highest industrialization potential are direct recycling techniques due to low costs and simple procedures. Concerning phase separation, froth flotation and ultrasound-assisted methods are discussed. The former divides black mass into pure anodic and cathodic materials, while ultrasonication is employed to physically detach active materials from foils or enhance binder degradation. As to materials recovery, several recent approaches show high recovery efficiency for different elements, mainly in leaching. The use of new organic acids, deep eutectic acids, and some salts are worth noting as leaching agents due to their low environmental impact. In addition, leaching methods assisted by ultrasound and microwave irradiation increase valuable metal recovery, reducing time consumption and the number of leaching reactants. As a part of the hydrometallurgy process, metallic ion purification is performed by solvent extraction and ion exchange, while selective precipitation can be achieved by specific chemical agents or electrochemical processes., Competing Interests: Declaration of competing interest The authors declare no known competing financial interests or personal relationships that may influence the work reported in this paper., (Copyright © 2023. Published by Elsevier B.V.)

Objective: In the bacterial etiology of severe acute infectious diarrhea, except that caused by Clostridioides difficile, most of them have an invasive character and antibiotic treatment will be necessary in specific situations. Shigella is a classic pathogen, in which it is crucial to know the sensitivity to different classic and alternative antimicrobials. The objective of this work was to analyze the presence of shigellosis and the rate of antibiotic resistance., Methods: A descriptive-retrospective study of the reports of shigellosis of stool cultures issued between January 2016 and April 2022 was conducted., Results: A total of 34 episodes (16 -47.1%- by Shigella sonnei) were observed, as of 2018. There were only 2 pediatric cases. The overall resistance rate to azithromycin, trimethoprim-sulfamethoxazole and ciprofloxacin was 52.9%, 64.7% and 44.1%, respectively. 26.5% were resistant to the 3 groups of antibiotics. There was a higher rate of resistance for S. sonnei. The emergence of resistance to cephalosporins in recent years stands out. Episodes of multidrug-resistant shigellosis were detected between 2020 (1 by S. flexneri) and 2022 (4 by S. sonnei)., Conclusions: The episodes of shigellosis are emerging in our environment with a higher rate of multi-resistance. In this context, current empirical treatments for acute enteroinvasive enteritis are at risk of failure, if necessary., (©The Author 2023. Published by Sociedad Española de Quimioterapia. This article is distributed under the terms of the Creative Commons Attribution-NonCommercial 4.0 International (CC BY-NC 4.0)(https://creativecommons.org/licenses/by-nc/4.0/).)

Background: Salmonella enterica serovar Typhimurium (S. Typhimurium) has become an important intestinal pathogen worldwide and is responsible for lethal invasive infections in populations at risk. There is at present an unmet need for preventive vaccines., Methods: IRTA GN-3728 genome was sequenced by Illumina and d-glutamate and d-glutamate/d-alanine knockout-auxotrophs were constructed. They were characterized using electron microscopy, growth/viability curves, reversion analysis, and motility/agglutination assays. Their potential as vaccine candidates were explored using two BALB/c mouse models for Salmonella infections: a systemic and an intestinal inflammation. Clinical signs/body weight and survival were monitored, mucosal lactoferrin and specific/cross-reactive IgA/IgG were quantified by enzyme-linked-immunosorbent assays and bacterial shedding/burden in fecal/tissues were evaluated., Results: The d-glutamate auxotroph, IRTA ΔmurI, is highly attenuated, immunogenic and fully protective against systemic infection. The IRTA ΔmurI Δalr ΔdadX double auxotroph, constructed to reinforce vaccine safety, showed a higher level of attenuation and was 100% effective against systemic disease. In the intestinal model, it proved to be safe, yielding a low-degree of mucosal inflammation, short-term shedding and undetectable invasiveness in the long-term, while eliciting cross-reactive fecal IgA/serum IgG against clinically relevant multidrug-resistant (MDR) S. Typhimurium strains. It also conferred protection against homologous oral challenge, and protected mice from local and extra-intestinal dissemination caused by one MDR strain responsible for an international outbreak of highly severe human infections. Additionally, oral vaccination promoted extended survival after lethal heterologous infection., Conclusion: This study yielded a very safe S. Typhimurium vaccine candidate that could be further refined for mucosal application against disease in humans., Competing Interests: Declaration of competing interest The authors declare that they have no competing interests., (Copyright © 2022. Published by Elsevier B.V.)

The aim of our study was to delineate an outbreak of gastroenteritis caused by Shigella flexneri and affecting sixteen persons between May and June 2014 in Bilbao, Spain. All patients exhibited symptoms after consuming kebab in the same kebab shop.The outbreak is described through the clinical cases, the microbiological and molecular genetic diagnosis, and the epidemiologic investigation. Minimum inhibitory concentrations for ampicillin, amoxicillin plus clavulanic acid, third and fourth generation cephalosporins, carbapenems, monobactams, aminoglycosides, fluoroquinolones, co-trimoxazole, colistin and tigecycline were measured. The S. flexneri strains were screened by PCR for TEM, SHV, CTX-M beta-lactamases and plasmidic AmpCs and aac(6')-Ib gene. Serotyping, pulsed field gel-electrophoresis, conjugation assay, plasmid sizing by S1 enzyme digestion and Southern blot hybridization were accomplished.All the S. flexneri isolates proved to be serotype 2 and produced extended-spectrum beta-lactamase (ESBL). Carbapenems, fluoroquinolones, tigecycline, colistin, and co-trimoxazole remained active antibiotics. All the strains harboured blaCTX-M-15 and blaOXA-1 genes. The strains hosted two high-molecular weight plasmids of 100 and 230 kb, respectively. According to the hybridization assay blaCTX-M-15 was located on the plasmid of 230 kb. The identical pulsotype verified the presence of outbreak.Remarkable, that one of the food handlers has travelled recently to Pakistan, where ESBL-producing Shigella strains had been reported previously. To the best of our knowledge, this is the first outbreak caused by CTX-M-15-expressing S. flexneri in Spain and as well as in Europe.

During the last decades, antimicrobial resistance (AMR) has become a global public health concern. Nowadays multi-drug resistance is commonly observed in strains of Vibrio cholerae , the etiological agent of cholera. In order to limit the spread of pathogenic drug-resistant bacteria and to maintain treatment options the analysis of clinical samples and their AMR profiles are essential. Particularly, in low-resource settings a timely analysis of AMR profiles is often impaired due to lengthy culturing procedures for antibiotic susceptibility testing or lack of laboratory capacity. In this study, we explore the applicability of whole genome sequencing for the prediction of AMR profiles of V. cholerae . We developed the pipeline CholerAegon for the in silico prediction of AMR profiles of 82 V. cholerae genomes assembled from long and short sequencing reads. By correlating the predicted profiles with results from phenotypic antibiotic susceptibility testing we show that the prediction can replace in vitro susceptibility testing for five of seven antibiotics. Because of the relatively low costs, possibility for real-time data analyses, and portability, the Oxford Nanopore Technologies MinION sequencing platform-especially in light of an upcoming less error-prone technology for the platform-appears to be well suited for pathogen genomic analyses such as the one described here. Together with CholerAegon, it can leverage pathogen genomics to improve disease surveillance and to control further spread of antimicrobial resistance., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2022 Fuesslin, Krautwurst, Srivastava, Winter, Liedigk, Thye, Herrera-León, Wohl, May, Fobil, Eibach, Marz and Schuldt.)

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.

Many regions around the world are suffering from water stress, and desalinated water and recycled water are seen as alternatives for meeting the water demand. However, high energy consumption and associated greenhouse gas emissions are some of the main environmental impacts. This is notable for many arid and semi-arid countries where desalination and water recycling are considered options for ensuring water resources availability. This research presents the incorporation of the quantification of greenhouse gas emissions generated during the operation of desalination and wastewater treatment plants in the assessment of water stress levels using the water stress indicator adopted by the 2030 Agenda for Sustainable Development. Chile was chosen as a case study, as it is a country where there is a considerable difference between the availability of conventional water sources and the water demand, and the electrical grid is fed mainly by fossil fuels. The methodology proposed allows calculating the indirect greenhouse gas emissions due to electrical consumption for the operation of desalination and wastewater treatment plants, and the direct greenhouse gas emissions coming from biological processes used in wastewater treatment plants. The results showed that Chilean arid climate zones will not experience water stress in the future at the regional level, mainly because of the installation of several desalination plants by 2030. Meanwhile, recycled water from the urban sector will slightly contribute to the reduction in the level of water stress in almost all Chilean regions by 2030. Moreover, desalination and wastewater treatment plant will contribute only between 0.34% and 0.75% of total greenhouse gas emitted in Chile by 2030. Therefore, the operation of these industrial systems for facing water scarcity problems in northern and central zones of Chile is a suitable alternative because it does not generate large environmental problems., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2021 Elsevier B.V. All rights reserved.)

An extensive multi-country outbreak of multidrug-resistant monophasic Salmonella Typhimurium infection in 10 countries with 150 reported cases, predominantly affecting young children, has been linked to chocolate products produced by a large multinational company. Extensive withdrawals and recalls of multiple product lines have been undertaken. With Easter approaching, widespread product distribution and the vulnerability of the affected population, early and effective real-time sharing of microbiological and epidemiological information has been of critical importance in effectively managing this serious food-borne incident.

Shortly after the declaration of the COVID-19 pandemic, governments around the world were urged to leave no population behind. Following a COVID-19 risk evaluation in a refugee and asylum seekers reception center, in September 2020, we considered the priorities of managing COVID-19 in these settings. We encourage actions on the following four fronts to reduce the COVID-19-associated burden among these vulnerable populations based on our interviews, observations, and recommendations: (i) decongestion, (ii) facilitated testing, (iii) screening for symptoms, and (iv) targeted public health and risk communication., Competing Interests: Declaration of Competing Interest The authors declared no conflicts of interest., (Copyright © 2022. Published by Elsevier Ltd.)