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[Objectives] The aim of this study was to assess the immunogenicity of SARS-CoV-2 available vaccines among people living with HIV (PLWH) after a complete vaccination scheme, and determine predictors of seroconversion., [Methods] This multicentre prospective cohort study included 420 PLWH who had received a standard immunization, either with mRNA or adenoviral-vectored COVID-19 vaccines. Antibody response was evaluated within 1 to 2 months after the last dose of the vaccine with a quantitative determination of antitrimeric spike protein-specific IgG antibodies and IgG neutralizing antibodies., [Results] Overall, 384 of 420 PLWH (91%) showed antibody response to vaccination. Seroconversion was observed in 308 of 326 individuals with cluster of differentiation 4 (CD4) counts ≥350 cells/mm3 (95%), 55 of 61 PLWH with 200 to 349 cells/mm3 (90%), and 21 of 33 PLWH with CD4 counts, [Discussion] HIV-related immunosuppression impairs the antibody response to SARS-CoV-2 vaccines. Specific vaccination schemes should be urgently tailored in this setting, particularly in patients with CD4 cell counts, This work was supported, in part, by the Spanish Network for AIDS Investigation (www.red.es/redes/inicio; RD16/0025/0040) as part of the Nacional I + D + I, ISCIII Subdirección General de Evaluación and the European Fund for Development of Regions. Juan Antonio Pineda received a research extension grant from the Programa de Intensificación de la Actividad de Investigación del Servicio Nacional de Salud Carlos III. Anaïs Corma-Gómez received a Río...

Nowadays, it is of utmost importance to use fully validated assays for molecular-based diagnosis. In the field of sexually transmitted disease (STD), Roche and Hologic provide assays for diagnosing Chlamydia trachomatis (CT), Neisseria gonorrhoeae (NG), Mycoplasma genitalium (MG), and Trichomonas vaginalis (TV). A total of 212 clinical samples were tested. Aptima® Combo 2 (detecting CT and NG), Aptima® M. genitalium and the Aptima® T. vaginalis on the Panther® system were compared to CoBAS® CT/NG and CoBAS® TV/MG running on the CoBAS® 6800 system. To solve the discrepancies, Allplex™ STI Essential assay (Seegene®) and/or Sanger DNA sequencing were used. The diagnostic performance was calculated by mean of the sensitivity and specificity parameters. Aptima® (sensitivity: 98.90%, specificity: 100%), CoBAS® (sensitivity 100%, specificity: 96.67%). The CoBAS® combo (CT/NG) failed detecting NG from an anal/rectum specimen, which is not included into the validated specimens of the assay. Aptima® combo 2 produced two false positives (CT and NG), not detected by the third tests. All the assays showed an optimal diagnostic capacity, meeting the requirements for IVD DNA-based assays. All products work optimally on automatic platforms, minimizing time and risk of contamination during handling.

The objective was to evaluate the accuracy of a single determination of IgM antibodies for hepatitis E virus (HEV) diagnosis in patients with acute hepatitis. A prospective study included patients with suspicion of HEV infection, defined as individuals with acute hepatitis showing negative results for serological and molecular markers of other hepatitis viruses. All patients were evaluated for hepatitis E virus infection, including both IgM antibodies and viral RNA determinations. Hepatitis E virus infection was defined as positivity for any of these markers. A total of 182 patients were included in the study, of whom 68 (37.4%) were diagnosed with HEV infection. Of these, 29 (42.6%) were positive for both IgM and HEV RNA, 25 (36.8%) were positive only for IgM antibodies, and 14 (20.6%) were positive only for HEV RNA. Considering only those individuals who were positive for IgM antibodies, 54 of the 68 total cases (79.4%) could be identified, showing a percentage of false-negative individuals of 20.6%. The diagnostic algorithm of hepatitis E virus infection in patients with acute hepatitis should include the determination of both IgM antibodies and HEV RNA because single sampling for IgM antibody determination led to an important proportion of misdiagnosed cases., This work was supported by the Ministerio de Sanidad (RD12/0017/0012) integrated into the Plan Nacional de I+D+I and cofinanced by the ISCIII-Subdirección General de Evaluación and the Fondo Europeo de Desarrollo Regional (FEDER); the Fundación para la Investigación en Salud (FIS) del Instituto Carlos III (research project grant number PI19/00864); and the Red de Investigación en SIDA de España ISCIII-RETIC (grant number RD16/0025/0034). A.R.-J. is the recipient of a Miguel Servet research contract by the Ministerio de Ciencia, Promoción, y Universidades of Spain (CP18/00111). M.F. is the recipient of a Sara Borrell research contract program by the Ministerio de Ciencia, Promoción, y Universidades of Spain (CD18/00091). A.R. is the beneficiary of Contratos para la Intensificación de la Actividad Investigadora en el Sistema Nacional de Salud by the Ministerio de Ciencia, Promoción, y Universidades of Spain (INT20-00028).

ObjectivesIn recent years, resistance in Mycoplasma genitalium (MG) to first-line (azithromycin) and second-line (moxifloxacin) treatment has been increasingly reported worldwide, however, no data regarding the south of Spain are available.MethodsTo determine resistance rates, MG-positive samples collected from June 2018 to June 2019 were analysed by sequencing the 23S rRNA and parC genes.ResultsA total of 77 patients (24 men having sex with men (MSM), 30 heterosexual men and 23 women) were included. Resistance-associated mutations against macrolide and fluoroquinolones were found in 36.4% (95% CI 25.7% to 48.1%) and 9.1% (95% CI 3.7% to 17.8%) of the patients, respectively. Being MSM and having had another STI in the last year were significantly associated with macrolide-resistant MG infection, while no associations were found with resistance to fluoroquinolones.ConclusionsTesting for resistance to first-line and second-line drugs against MG should be recommended for the general population and mandatory for the MSM population. We suggest that empiric azithromycin use for STI management should be avoided.

Pathogens causing sexually transmitted diseases (STDs) include viruses, bacteria, and parasites. The ability to rapidly and efficiently detect these pathogens in a single reaction still remains a health challenge. The aim of this study was to evaluate the clinical reliability and accuracy of the STD Direct Flow Chip Kit (Vitro, IVD-EC approved), which can simultaneously detect up to 9 different species of STD pathogens at once. This kit enables direct analysis-direct-PCR-of clinical specimens (urine, semen, endocervical, urethral, nasopharyngeal, and perianal swabs) without DNA purification for the following pathogens: Chlamydia trachomatis (serovars A-K and L1-L3), Haemophilus ducreyi, Herpes Simplex Virus (Types I and II), Mycoplasma genitalium, Mycoplasma hominis, Neisseria gonorrhoeae, Treponema pallidum, Trichomonas vaginalis, and Ureaplasma. The Anyplex™ II STI-7 Detection Kit (Seegene, IVD-EC) was used as the reference's method. Existing discordances were resolved using either a third molecular assay or DNA sequencing. Clinical performance was evaluated at two different stages: (i) from purified DNA of three hundred and fifty-eight clinical specimens with a diagnostic sensitivity (SE) and specificity (SP) of 99.4% and 100%, respectively, and an agreement of 99% (kappa index, κ = 0.97) with the reference's method and; (ii) by direct-PCR from six hundred and thirty-three specimens rendering SE, SP, and agreement values of 98.4%, 99.9%, and 98.0% (κ = 0.95), respectively. The STD Direct Flow Chip Kit constitutes a promising alternative to routine procedures in diagnostic, allowing direct analysis of specimens and enabling the detection of a broad panel of pathogens.

BackgroundWhether people living with HIV (PLWH) are at greater risk of acquiring SARS-CoV-2 infection is currently unknown. Prospective serologic studies may allow seroincidence analyses, where all infections are accurately identified. Because of this, we evaluated the incidence of and associated factors with SARS-CoV-2 infection in PLWH in Southern Spain.MethodsThis was a prospective cohort study including PLWH from a University Hospital in Southern Spain. Patients were enrolled if 1) they had attended as outpatients our Unit from August 1st, 2019 to February 8th, 2020; 2) had two subsequent evaluations from February 9th, 2020 to February 15th, 2021. Serum antibodies against SARS-CoV-2 were determined in baseline and intra-pandemic samples.Results710 PLWH were included in the study. Of them, 46 [6.5%, 95% confidence interval (95% CI): 4.8%-8.5%] patients developed SARS-CoV-2 infection. Between May 18th and November 29th, 2020, the rate of seroconversion was 5.3% (95% CI: 3.1%-9%) for the general population in the area of Seville and 2.3% (95% CI: 1.3%-3.6%) for PLWH in this study (p=0.001). After multivariate analysis, adjusted by age and sex, active tobacco smoking was the only factor independently associated with lower risk of SARS-Cov-2 infection (Incidence rate ratio 0.35, 95% CI: 0.18-0.68, p=0.002).ConclusionsThe incidence of SARS-CoV-2 infection among PLWH in Southern Spain during the ongoing pandemic was lower than that reported for the general population in the same area. Tobacco smoking was the only factor independently associated with a lower risk of incident SARS-CoV-2 infection.SummaryThe incidence of SARS-CoV-2 infection among people living with HIV is lower than that of general population in Southern Spain. Active tobacco smoking could be associated with a lower risk of developing COVID-19.

Whether people living with HIV (PLWH) are at greater risk of acquiring SARS-CoV-2 infection is currently unknown. Prospective serologic studies may allow seroincidence analyses, where all infections are accurately identified. Because of this, we evaluated the incidence of associated factors with and the clinical outcome of SARS-CoV-2 infection in PLWH in Southern Spain. This prospective cohort study included PLWH from a Tertiary University Hospital in Southern Spain. Patients were enrolled in the study if (1) they had attended as outpatients our Unit from 1 August 2019 to 8 February 2020 and (2) had two subsequent evaluations from 9 February 2020 to 4 March 2021. SARS-CoV-2 infections were diagnosed by PCR, antigen detection or serology. Seven hundred and nine PLWH were included in the study. Of them, 55 [7.8%, 95% confidence interval (95% CI) 5.9%-9.9%] patients developed SARS-CoV-2 infection. Between 18 May and 29 November 2020, the rate of seroconversion was 5.3% (95% CI: 3.1%-9.0%) for the general population in the area of Seville and 2.3% (95% CI: 1.3%-2.6%) for PLWH in this study (p = .001). After multivariable analysis, adjusted by age, sex, and risk factors for HIV infection, active tobacco use and CDC stage, active tobacco smoking was the only factor independently associated with lower risk of SARS-Cov-2 infection [Incidence rate ratio: 0.29 (95% CI 0.16-0.55) p < .001]. In conclusion, the incidence of SARS-CoV-2 infection among PLWH in Southern Spain during the ongoing pandemic was lower than that reported for the general population in the same area. This work was supported in part by the Instituto de Salud CarlosIII (Project ‘PI16/01443’), integrated in the national I+D+i 2013–2016andco-fundedbytheEuropeanUnion(ERDF/ESF,‘Investinginyour future’), by the Spanish Network for AIDS investigation (RIS)(www.red.es/redes/inicio)(RD16/0025/0010,RD16/0025/0040),asapartoftheNacionalI+D+I,ISCIIISubdirecciónGeneraldeEvaluaciónand the European Fund for Development of Regions (FEDER). JuanA.PinedareceivedaresearchextensiongrantfromtheProgramadeIntensificacióndelaActividaddeInvestigacióndelServicioNacionaldeSaludCarlosIII(I3SNS).FedericoGarciareceivedaresearchextensiongrantfromtheProgramadeIntensificacióndelaActividaddeInves-tigacióndelServicioAndaluzdeSalud.AnaïsCorma-GomezreceivedaRíoHortegagrantfromtheInstitutodeSaludCarlosIII(grantnum-ber CM19/00251). Funding for open access charge: Universidad deMálaga/CBUA

Importance: Information about risk factors for SARS-CoV-2 infection and COVID-19 outcome among people living with human immunodeficiency virus (PLWH) is limited. Furthermore, the impact of smoking on the risk of developing COVID-19 is unclear and has been a matter of controversy.Objective: To evaluate the relationship between current tobacco smoking and the probability of SARS-CoV-2 infection acquisition among PLWH.Design: Retrospective nested case-control study performed within a cohort of 702 PLWH.Settings: University Hospital in Spain (February-October 2020).Participants: Cases were the 18 patients from this cohort with a confirmed diagnosis of COVID-19. Control group included the remaining 684 PLWH without a diagnosis of COVID-19.Exposure: Active tobacco smoking vs. non-smoking, using self-reported electronic health record data.Outcome and Measure: Development of COVID-19. Diagnosis of COVID-19 was established by the detection of SARS-CoV-2 RNA or antigen by PCR or EIA, respectively, or by the presence of plasma SARS-CoV-2 antibodies by ECLIA.Results: In 11 (61.1%) of the 18 patients, COVID-19 diagnosis was based on PCR, 4 (22.2%) on antigen and 3 (16.7%) on serology determinations. In the control group, 380 (54.1%) individuals were also tested. Regarding clinical spectrum, 2 (11.1%) patients were asymptomatic, 12 (66.6%) had upper-respiratory tract infections and 2 (11.1%) had viral pneumonia and one (5.6%) individual showed extra respiratory symptoms. One (5.6%) patient developed severe pneumonia and died. Three hundred and sixty-nine (52.6%) individuals were active smokers, 2 (11.1%) at the COVID-19 diagnosis and 367 (54.1%) in control group (p=0.001). In the bivariate analysis, only tobacco smoking was associated with a greater risk of COVID-19 [unadjusted OR=9.41 95% Confidence Interval (95% CI, 2.15-41.24); p=0.003]. Multivariate analysis, adjusted by sex, tobacco smoking and Charlson index, showed that tobacco smoking was independently related to a higher probability of SARS-CoV2 infection [aOR=9.42 (95% CI 2.15-41.33), p = 0.003].Conclusion and Relevance: Active tobacco smoking is associated with a lower incidence of SARS-CoV-2 infection in PLWH. The underlying mechanisms by which tobacco smoking may protect against SARS-CoV-2 infection should be further investigated. In the meanwhile, public-facing messages about a protective effect of tobacco smoking on the risk of COVID-19 must be avoided.

Objective and Design: The incidence of SARS-CoV-2 infection among people living with HIV (PLWH) has been estimated on the basis of reported symptomatic clinical cases. However, as asymptomatic cases are common and there have been limitations of health care systems for COVID-19 microbiological diagnosis, these estimations may be misleading. The availability of reliable serology for the diagnosis of COVID-19 may overcome this drawback. This study was carried out in order to reveal the actual incidence of SARS-CoV-2 infection in PLWH in Southern Spain.Methods: This is a prospective cohort study including HIV infected patients from the Unit of Infectious Diseases of a university hospital in Seville, Southern Spain. Patients were enrolled in the study if 1) they had attended the outpatient clinic from September 1st to December 31st, 2019 (baseline), and 2) had a subsequent evaluation from March 1st to June 30th, 2020 (intra-pandemic). Serum antibodies against SARS-CoV-2 were determined in baseline and intra-pandemic samples.Results: 326 patients were included in the study. Of them, 4 (1.25% [95% CI: 0.3%-3.1%]) developed COVID-19. One patient developed bilateral pneumonia and died. The remaining three showed mild respiratory symptoms suggesting COVID-19. No asymptomatic SARS-CoV-2 infection was observed in this study. No patient with COVID-19 was tobacco smoker. The incidence of COVID-19 among non-smokers was 2.5% (95% CI [0.6%-6%], p=0.057 versus smokers).Conclusions: The incidence of COVID-19 among PLWH in our area was low and similar to that observed in the general population. The frequency of asymptomatic cases might be lower than in patients without HIV infection. Tobacco smoking could be associated to a lower incidence of COVID-19.

The extensive use of essential oil components in an increasing number of applications can substantially enhance exposure to these compounds, which leads to potential health and environmental hazards. This work aimed to evaluate the toxicity of four widely used essential oil components (carvacrol, eugenol, thymol, vanillin) using the in vivo model Caenorhabditis elegans. For this purpose, the LC

The vaginal microbiota has importance in preserving vaginal health and defending the host against disease. The advent of new molecular techniques and computer science has allowed researchers to discover microbial composition in depth and associate the structure of vaginal microbial communities. There is a consensus that vaginal flora is grouped into a restricted number of communities, although the structure of the community is constantly changing. Certain Community-State Types (CSTs) are more associated with poor reproductive outcomes and sexually transmitted diseases (STDs) meanwhile, CSTs dominated by Lactobacillus species—particularly Lactobacillus crispatus—are more related to vaginal health. In this work, we have reviewed how modifiable and non-modifiable factors may affect normal vaginal microbiota homeostasis—including sexual behavior, race or ethnicity, and hygiene. Special interest has been given to how the use of probiotics, diet intake, and use of hormone replacement therapies (HRTs) can potentially impact vaginal microbiota composition.

In recent years, resistance inTo determine resistance rates, MG-positive samples collected from June 2018 to June 2019 were analysed by sequencing the 23S rRNA and parC genes.A total of 77 patients (24 men having sex with men (MSM), 30 heterosexual men and 23 women) were included. Resistance-associated mutations against macrolide and fluoroquinolones were found in 36.4% (95% CI 25.7% to 48.1%) and 9.1% (95% CI 3.7% to 17.8%) of the patients, respectively. Being MSM and having had another STI in the last year were significantly associated with macrolide-resistant MG infection, while no associations were found with resistance to fluoroquinolones.Testing for resistance to first-line and second-line drugs against MG should be recommended for the general population and mandatory for the MSM population. We suggest that empiric azithromycin use for STI management should be avoided.

Sexually transmitted infections (STIs) remain a worldwide problem and a severe threat to public health. The purpose of this study was to compare Aptima® Assays (Hologic®) and the Allplex™ STI Essential Assay (Seegene®) for the simultaneous detection of Chlamydia trachomatis, Neisseria gonorrhoeae, Trichomonas vaginalis and Mycoplasma genitalium in clinical practice. The Aptima® assays (Hologic®) are based on a transcription-mediated amplification (TMA) method. The Allplex™ STI Essential assay (Seegene®) is based on a multiplex Real-Time PCR (RT-PCR) method. A total of 622 clinical samples from different anatomical sites were tested using both methods. A total of 88 (14.1%) and 66 (10.6%) positive samples were found for any of the TMA assays used and for the RT-PCR assay, respectively. Aptima® assays showed a slightly higher rate of positive results for all pathogens except for T. vaginalis, the results of which were similar to those obtained with Allplex™. The most commonly detected pathogen was C. trachomatis (37 samples; 5.9% using TMA assays) and the anatomical site with the highest prevalence of microorganisms was a non-urogenital site, the pharynx (27 positive samples; 4.3%). Using the Aptima® assays as reference method, the comparison showed that the average specificity of multiplex RT-PCR was 100.0% for the four pathogens. However an average sensitivity of 74.5% was observed, showing 95.2% (CI95%; 93.6–96.9) of overall concordance (κ = 0.80). In conclusion, the Aptima® assays show a higher sensitivity on a wide range of sample types compared to the Allplex™ assay.

Graham J Etherington should be included in the author byline. He should be listed as the sixth author and his contribution is: Genome sequencing and SNPs identification in the mutant plant. His affiliations are 1: The Sainsbury Laboratory, Norwich Research Park, Norwich, United, Kingdom, and 2: The Genome Analysis Centre, Norwich Research Park, Norwich, United Kingdom. The correct citation is: Ramirez V, Gonzalez B, Lopez A, Castello MJ, Gil MJ, Etherington GL, et al. (2015) Loss of a Conserved tRNA Anticodon Modification Perturbs Plant Immunity. PLoS Genet 11(10): e1005586. doi: 10.1371/journal.pgen.1005586

[EN] The microbiological and chemical processes are the main responsible for deterioration of fresh fish. Therefore, it is essential to avoid these processes by applying good manufacturing practices during fish handling, distribution and storage. Objective: The aim of this paper was to evaluate the physico-chemical and microbial changes in commercial tilapia (Oreochromis niloticus) during cold storage in order to establish the shelf life since its arrival at the supermarket. Methods: 27 aquacultured tilapia specimens were analyzed at 0, 2, 4, and 7 days of storage at 4ºC. Measurements of texture, color, water holding capacity, total volatile basic nitrogen (TVB-N), thiobarbituric acid index, ATP-related compounds, as well as microbial analyses were carried out. Results: The TVB-N content was high at the beginning of the study, remaining stable during the storage. Lipid oxidation of samples was minimum, so this process did not contribute to the fish spoilage. It was observed the breakdown of inosine-5’- monophosphate (IMP) into Ino (inosine), and Ino into Hx (hypoxantine). The texture parameters and colour underwent changes as a consequence of the fish spoilage. Low microbial counts were observed at day 0, but Enterobacteriaceae and mesophilic counts gradually increased throughout storage. Conclusions: The K1-value showed the progressive spoilage of the fish during the cold storage. The decrease of hardness and firmness confirmed the loss of quality throughout the time of study. The low microbial counts at the beginning of the study demonstrated the good quality of the tilapia; however, the increase of the mesophilic counts at the end of the studied period showed that tilapia was not fit for human consumption at day 7., [ES] Antecedentes: Los procesos microbiológicos y químicos son los principales responsables del deterioro del pescado fresco. Por tanto, es esencial evitar estos procesos aplicando buenas prácticas de fabricación durante la manipulación, distribución y almacenamiento del pescado. Objetivo: El objetivo de este trabajo fue evaluar los cambios físico-químicos y microbiológicos en tilapia comercial (Oreochromis niloticus) durante el almacenamiento refrigerado con el fin de establecer la vida útil desde su llegada al supermercado. Métodos: 27 especímenes de tilapia fueron analizados a día 0, 2, 4 y 7 de almacenamiento a 4 ºC. Se llevaron a cabo medidas de textura, color, capacidad de retención de agua, nitrógeno básico volátil total (N-BVT), índice del ácido tiobarbitúrico, compuestos relacionados con el ATP, así como análisis microbianos. Resultados: El contenido en N-BVT fue alto al principio del estudio, siendo estable durante el almacenamiento. La oxidación lipídica de las muestras fue mínima, por lo que este proceso no contribuyó al deterioro del pescado. Se observó la degradación de inosina-5’- monofosfato (IMP) a Ino (inosina), y de Ino a Hx (hipoxantina). Los parámetros de textura y color sufrieron cambios como consecuencia del deterioro del pescado. Se observaron bajos recuentos microbianos a día 0, pero los recuentos de Enterobacteriaceae y de mesófilos aumentaron gradualmente durante el almacenamiento. Conclusiones: El valor K1 mostró el deterioro progresivo del pescado durante el almacenamiento en refrigeración. La disminución de dureza y firmeza confirmó la pérdida de calidad a lo largo del tiempo de estudio. Los bajos recuentos microbianos al principio del estudio demostraron la buena calidad de la tilapia; sin embargo, el aumento de los recuentos de mesófilos al final del periodo estudiado mostraron que la tilapia no era adecuada para el consumo humado a día 7.

The authors would like to correct Fig 2. The figure contains two errors. In Fig 2A, the sbt3.4–2 representative picture was duplicated from Figure 5A (NPR1-H). These errors occurred during assembly of the final figure. Additionally, in Fig 2C, the middle Ponceau panel (ssbt3.3–1 experiment) was duplicated in the bottom Ponceau panel (sbt3.3–2 experiment). The authors have provided a corrected Fig 2 here. The authors confirm that these changes do not alter their findings. The authors have provided raw, uncropped blots as Supporting Information.