A Potential Nervous Necrosis Virus (NNV) Live Vaccine for Sole Obtained by Genomic Modification.

Viral Encephalopathy and Retinopathy (VER) is a neurological infectious fish disease that causes vacuolization and necrosis in the central nervous system, which lead to swimming abnormalities and, generally, host death in the early stages of development. VER is caused by the Nervous Necrosis Virus (NNV), a non-enveloped virus with a bisegmented and positive-stranded (+) RNA genome. The largest segment (RNA1) codes for viral polymerase while capsid protein is encoded by RNA2. The aim of this study was to explore the potential of a reverse-engineered RGNNV/SJNNV strain that harbors mutations in both 3'NCRs (position 3073 of RNA1 and 1408 and 1412 of RNA2) as an attenuated live vaccine for sole. The attenuation of this strain was confirmed through experimental infections in sole at 22 °C. Vaccination trials were performed by bath, intramuscular, and intraperitoneal injection, at two temperatures (18 and 22 °C). Our results indicate the improved survival of vaccinated fish and delayed and poorer viral replication, as well as an overexpression of immune response genes linked to T cell markers (cd4 and cd8), to an early inflammatory response (tlr7 and tnfα), and to antiviral activity (rtp3 and mx). In conclusion, our study indicates that the attenuated strain is a good vaccine candidate as it favors sole survival upon infection with the wt strain while inducing a significant immune response.

Water-in-oil adjuvant challenges in fish vaccination: An experimental inactivated adjuvanted vaccine against betanodavirus infection in Senegalese sole.

The extensive growth of intensive fish farming has led to a massive spread of infectious diseases. Nervous necrosis virus (NNV) is the causative agent of the viral encephalo- and retinopathy disease which has become a major threat for fish farming all over the globe. The devastating mortality rates recorded in disease outbreaks, especially when infected specimens are at early stages of development, have a high economic impact on the sector. Currently, vaccines are the most cost-effective preventing tool in the fight against viruses. Inactivated vaccines have the advantage of simplicity in their development at the same time as present the antigen in a similar manner than the natural infection in the host. Nevertheless, they usually trigger weaker immune responses needing adjuvants to boost their effectiveness. In this work, we have intraperitoneally vaccinated Senegalese sole juveniles (Solea senegalensis) with a previously designed inactivated vaccine against NNV based on binary ethylenimine (BEI), mixed or not with an oil-adjuvant. Our results demonstrated the potential activation of different immune pathways when the vaccine was administered alone compared to the oil-adjuvanted vaccine, both resulting in an equivalent partial improvement in survival following a NNV challenge. However, whilst the vaccine alone led to a significant increase in specific antibodies, in the adjuvanted version those antibodies were kept basal although with a slight improvement in their neutralization capacity. At transcriptional level, neither vaccine (adjuvanted or not) triggered the immune system activation during the vaccination period. However, after NNV infection, the BEI-inactivated vaccines alone and oil-adjuvanted both elicited the stimulation of antiviral responsive genes (rtp3, herc4), antigen presentation molecules (mhcii) and T-cell markers (cd8a) in the head-kidney. Additionally, the oil-adjuvanted vaccine appears to stimulate mediator cytokines (il6) and B-cell markers (ight and ighm). Surprisingly, when the adjuvant was administered alone, fish showed the highest survival rates concomitantly with a lack of NNV-IgM production, pointing to the possible induction of different immune pathways than the B-cell responses via antibodies by the adjuvant. Since this combined vaccine did not succeed in the full extension of protection against the pathogen, further studies should be performed focusing on unravelling the molecular mechanisms through which adjuvants trigger the immune response, both independently and when added to a vaccine antigen.

Women's fear of childbirth during pregnancy: A concept analysis.

PURPOSE: To map and analyze the concept of women's fear of childbirth (FOC) during pregnancy and to develop a new nursing diagnosis (ND). METHODS: Concept analysis based on Walker and Avant's methodology. An integrative literature search was conducted in Scopus, Web of Science, and CINAHL Complete databases, including peer-reviewed articles published in English from January 1981 to October 2022. FINDINGS: Thirty-one studies were included in the review. Women's FOC during pregnancy comprises three central categories: attributes, antecedents, and consequences. Nine main characteristics (attributes) are related to one or more of eight prior conditions (antecedents) that have the potential to exert negative or favorable impacts (consequences), depending on whether coping strategies have been a part of a fearful woman's pregnancy. These elements and relationships among them should be viewed as dynamic. CONCLUSIONS: Women's FOC during pregnancy is a subjective, multidimensional, and dynamic concept that benefits prenatal care responses. A conceptual matrix on prenatal care will consider the factors underlying women's FOC and their potential mechanisms, how fearful pregnant women respond, the aspects that shape FOC, and how it influences outcomes when supporting pregnant women with FOC. Therefore, the study findings can be used to inform ways of identifying fearful women or as a basis for interventions to manage fears and enhance the confidence of pregnant women for childbirth. IMPLICATIONS FOR NURSING PRACTICE: The results of this study help nurses, OB nursing practitioners, nurse-midwives, and midwives to identify the condition of women's FOC during pregnancy and guide nursing therapeutic interventions to decrease the negative effects on women's health and well-being. Therefore, there are valuable reasons to propose using "fear of childbirth" as a ND in the class "coping responses" within domain coping/stress tolerance, contributing to refining the NANDA-I taxonomy.

OBJETIVO: Mapear e analisar o conceito de medo do parto em mulheres durante a gravidez e desenvolver um novo diagnóstico de enfermagem. MÉTODOS: Análise de conceito com base na metodologia de Walker e Avant. Foi realizada uma pesquisa integrativa da literatura nas bases de dados Scopus, Web of Science e CINAHL Complete e incluiu artigos revistos por pares, publicados em inglês, de janeiro de 1981 a outubro de 2022. RESULTADOS: Trinta e um estudos foram incluídos na revisão. O medo do parto em mulheres durante a gravidez é composto por três categorias centrais: atributos, antecedentes e consequências. Nove características principais (atributos) estão relacionadas com uma ou mais das oito condições anteriores (antecedentes) que têm o potencial de exercer um impacto negativo ou favorável (consequências), dependendo se estratégias de coping fizeram parte da gravidez de uma mulher com medo. Estes elementos e as relações entre eles devem ser vistos como dinâmicos. CONCLUSÕES: O medo do parto em mulheres durante a gravidez é um conceito subjetivo, multidimensional e dinâmico que beneficia de respostas pelos cuidados pré-natais. Uma matriz conceptual sobre cuidados pré-natais considerará os fatores subjacentes ao medo do parto nas mulheres e os seus potenciais mecanismos, como as mulheres grávidas com medo respondem, os aspetos que moldam o medo do parto, e como influenciam os resultados quando as mulheres grávidas com medo do parto são apoiadas. Portanto, os resultados do estudo podem ser usados para identificar mulheres com medo ou desenvolver intervenções que permitam gerir os medos e aumentar a confiança das mulheres grávidas para o parto. IMPLICAÇÕES PARA A PRÁTICA DE ENFERMAGEM: Os resultados deste estudo ajudarão os enfermeiros de atenção à saúde materna a identificar a condição do medo do parto nas mulheres durante a gravidez e orientar intervenções terapêuticas de enfermagem, no sentido de diminuir os efeitos negativos do medo na saúde e no bem-estar das mulheres. Portanto, há razões relevantes para propor a utilização do "medo do parto" como um diagnóstico de enfermagem na classe "coping responses" do domínio "coping/stress tolerance", contribuindo para o refinamento da taxonomia NANDA-I.

Designing and Validation of a Droplet Digital PCR Procedure for Diagnosis and Accurate Quantification of Nervous Necrosis Virus in the Mediterranean Area.

The viral nervous necrosis virus (VNNV) is the causative agent of an important disease affecting fish species cultured worldwide. Early and accurate diagnosis is, at present, the most effective control and prevention tool, and molecular techniques have been strongly introduced and accepted by official organizations. Among those, real-time quantitative polymerase chain reaction (rt-qPCR) is nowadays displacing other molecular techniques. However, another PCR-based technology, droplet digital PCR (ddPCR), is on the increase. It has many advantages over qPCR, such as higher sensitivity and more reliability of the quantification. Therefore, we decided to design and validate a protocol for the diagnosis and quantification of SJ and RG type VNNV using reverse transcription-ddPCR (RT-ddPCR). We obtained an extremely low limit of detection, 10- to 100-fold lower than with RT-qPCR. Quantification by RT-ddPCR, with a dynamic range of 6.8-6.8 × 104 (SJ type) or 1.04 × 101-1.04 × 105 (RG type) cps/rctn, was more reliable than with RT-qPCR. The procedure was tested and validated in field samples, providing high clinical sensitivity and negative predictive values. In conclusion, we propose this method to substitute RT-qPCR protocols because it exceeds the expectations of qPCR in the diagnosis and quantification of VNNV.

Recombinant viral hemorrhagic septicemia virus with rearranged genomes as vaccine vectors to protect against lethal betanodavirus infection.

The outbreaks of viral hemorrhagic septicemia (VHS) and viral encephalopathy and retinopathy (VER) caused by the enveloped novirhabdovirus VHSV, and the non-enveloped betanodavirus nervous necrosis virus (NNV), respectively, represent two of the main viral infectious threats for aquaculture worldwide. Non-segmented negative-strand RNA viruses such as VHSV are subject to a transcription gradient dictated by the order of the genes in their genomes. With the goal of developing a bivalent vaccine against VHSV and NNV infection, the genome of VHSV has been engineered to modify the gene order and to introduce an expression cassette encoding the major protective antigen domain of NNV capsid protein. The NNV Linker-P specific domain was duplicated and fused to the signal peptide (SP) and the transmembrane domain (TM) derived from novirhabdovirus glycoprotein to obtain expression of antigen at the surface of infected cells and its incorporation into viral particles. By reverse genetics, eight recombinant VHSVs (rVHSV), termed NxGyCz according to the respective positions of the genes encoding the nucleoprotein (N) and glycoprotein (G) as well as the expression cassette (C) along the genome, have been successfully recovered. All rVHSVs have been fully characterized in vitro for NNV epitope expression in fish cells and incorporation into VHSV virions. Safety, immunogenicity and protective efficacy of rVHSVs has been tested in vivo in trout (Oncorhynchus mykiss) and sole (Solea senegalensis). Following bath immersion administration of the various rVHSVs to juvenile trout, some of the rVHSVs were attenuated and protective against a lethal VHSV challenge. Results indicate that rVHSV N2G1C4 is safe and protective against VHSV challenge in trout. In parallel, juvenile sole were injected with rVHSVs and challenged with NNV. The rVHSV N2G1C4 is also safe, immunogenic and efficiently protects sole against a lethal NNV challenge, thus presenting a promising starting point for the development of a bivalent live attenuated vaccine candidate for the protection of these two commercially valuable fish species against two major diseases in aquaculture.

Midwives' interventions for reducing fear of childbirth in pregnant women: a scoping review.

OBJECTIVE: The objective of this review was to map and analyze midwives' interventions for reducing fear of childbirth in pregnant women. INTRODUCTION: Fear of childbirth is a phenomenon negatively affecting women's health and well-being before and during pregnancy. Over the past few decades, there has been growing research interest in interventions for reducing fear of childbirth in pregnant women. One of the challenges in midwifery care is to provide an appropriate model of care for pregnant women with fear of childbirth. Further research efforts are needed to identify midwives' interventions for reducing fear of childbirth in pregnant women and to examine their characteristics. INCLUSION CRITERIA: This scoping review considered studies that included midwives' interventions for reducing fear of childbirth in pregnant women. Specifically, interventions were led and/or implemented by midwives during the antenatal period, and integrating all possible midwifery practice settings. Quantitative, qualitative, and mixed methods studies were included. This review also considered systematic reviews, text and opinion papers, and conference abstracts. METHODS: The JBI methodology for conducting scoping reviews was used. Published and unpublished literature in English, Portuguese, and Spanish from January 1981 to October 2020 was included. MEDLINE (PubMed), CINAHL Complete, APA PsycINFO, Scopus, Embase, Web of Science, SciELO, MedicLatina, Academic Search Complete, ERIC, Psychology and Behavioral Sciences Collection, and the Cochrane Library databases were searched. Searches for gray literature were also undertaken on the Repositório Científico de Acesso Aberto de Portugal, ProQuest Dissertations and Theses, British Library EThOS, OvidSP Resource Center, Banco de Teses da CAPES, and OpenGrey. A three-step search strategy was followed, and the Preferred Reporting Items for Systematic Reviews and Meta-Analysis extension for Scoping Reviews checklist was used. Two independent reviewers extracted the data using a data extraction tool developed specifically for this scoping review. RESULTS: A total of 3704 articles were identified and screened, of which 34 articles were included. The majority of studies had been published in the past 10 years (88%) in Scandinavian countries or Australia (79%). Several midwives' antenatal interventions were found, such as midwife-led team models of care. Midwives played a facilitator role that varied across the included studies. In 20 studies (59%), midwives led and implemented the interventions alone ( n   =  13; 38%) or with the participation of other health professionals ( n   =  7; 21%). In the remaining 14 studies (41%), midwives were part of a multidisciplinary team that included different health professionals (mainly obstetricians and psychologists) who had been involved in delivering interventions alongside midwives or with minor participation from midwives. Counseling ( n   =  12; 35%) and psychoeducation ( n   =  8; 24%) were the most common midwife interventions for reducing fear of childbirth in pregnant women. CONCLUSIONS: Midwives working across their full scope of practice play a pivotal role in reducing fear of childbirth, which may explain the variety of midwives' antenatal interventions. Reducing fear of childbirth in pregnant women and promoting normal childbirth as a positive experience are key features of midwives' interventions, which should include women's empowerment measures. Evidence-based midwife-led intervention programs for pregnant women with fear of childbirth should be designed and tested to improve clinical practice as well as women's reproductive outcomes and perinatal experiences.

Prevalence and predictive factors for fear of childbirth in pregnant Portuguese women: A cross-sectional study.

OBJECTIVE: To measure the prevalence of the fear of childbirth (FOC) and determine which factors predict severe FOC among pregnant Portuguese women. METHODS: An online cross-sectional study among pregnant Portuguese women aged ≥ 20 years who were recorded using a convenience sampling. Self-administered questionnaires were used for data collection: socio-demographic and obstetric questionnaire and European Portuguese version of Wijma Delivery Expectancy Questionnaire-version A (WDEQ-A). Data of 669 participants were collected successfully from June 9 to October 30, 2019. Predictive factors for severe FOC were investigated using a multivariate logistic regression analysis. Odds ratios (OR) and 95% confidence intervals were calculated. RESULTS: The prevalence of severe FOC (WDEQ-A ≥ 85) among pregnant Portuguese women was 10%. Severe FOC was significantly associated with lower educational level, single/divorced marital status, and negative previous childbirth experience. Multivariate logistic regression analysis indicated that being single or divorced and having a negative previous childbirth experience were predictive variables for severe FOC. CONCLUSION: Pregnant Portuguese women have FOC, although with varying severity. The data suggest that marital status and women's perceptions of previous childbirth experience may be useful variables to predict severe FOC. Further research for extending the predictive factors of FOC should be refined. The results are clinically relevant for midwifery care, as they should be used in the sense of early identification of fearful pregnant women to provide adequate support strategies to reduce FOC.

Nervous Necrosis Virus (NNV) Booster Vaccination Increases Senegalese Sole Survival and Enhances Immunoprotection.

A re-immunization programme has been tested to improve the protective response elicited in sole by a previously developed BEI-inactivated betanodavirus vaccine. The vaccine was prepared using a reassortant RGNNV/SJNNV strain which is highly pathogenic for sole, and vaccination assays were performed by intraperitoneal injection. Experimental design included a prime- and a booster-vaccination group, which consisted of individuals that received a second vaccine injection at 30 days post vaccination), and their respective controls. A month after prime/booster vaccination, fish were challenged by intramuscular injection with the homologous NNV strain. Samples were collected at different times post vaccination and post challenge to assess the immune response and viral replication. Booster dose enhanced the protection against NNV infection because a significant increase in survival was recorded when compared with prime-vaccinated individuals (relative percent survival 77 vs. 55). In addition, a clear decrease in viral replication in the brain of challenged sole was observed. During the immune induction period, no differences in IgM production were observed between prime- and booster-vaccinated fish, and the expression of the antigen presenting cells (APC)-related molecule MHC class II antigen was the only differential stimulation recorded in the re-immunized individuals. However, a significant upregulation of mhcII and the lymphocytes T helper (Th) marker cd4 was observed after the challenge in the booster-vaccinated group, suggesting these cells play a role in the protection conferred by the booster injection. In addition, after viral infection, re-immunized fish showed specific and neutralizing antibody production and overexpression of other immune-related genes putatively involved in the control of NNV replication.

Differential Nervous Necrosis Virus (NNV) Replication in Five Putative Susceptible Cell Lines.

Viral encephalopathy and retinopathy caused by nervous necrosis virus (NNV), is one of the most threatening viral diseases affecting marine fish worldwide. In vitro propagation of NNV strains is essential for the design of effective control measures. In the present study we analysed both the susceptibility and the permissiveness of five fish cell lines (E-11, GF-1, SAF-1, DLB-1, and SaB-1) to three NNV strains (one RGNNV, one SJNNV, and one reassortant RGNNV/SJNNV). E-11 and DLB-1 were demonstrated to be highly susceptible to NNV strains, with average adsorption efficiency (AE) values higher than 90%. SAF-1 also showed high susceptibility (AE 88%), whereas GF-1 can be regarded as moderately susceptible (AE around 50%). On the contrary, SaB-1 can be considered a poorly susceptible cell line (AE values below 20%). E-11 and GF-1 cell lines provided the highest production rates for RGNNV and RG/SJ (around 103) and both cell lines can be regarded as fully permissive for these viral types. However, the SJNNV production rate in GF-1 was only 17.8 and therefore this cell line should be considered semi-permissive for this genotype. In SAF-1 cells, moderate viral replication was recorded but differences in intracellular and extracellular production suggest that viral progeny was not efficiently released. In DLB-1 and SaB-1 the final viral titres obtained in E-11 were lower than those of the inoculum. However, RNA1 synthesis values seem to indicate that RGNNV replication in DLB-1 and SAF-1 could have been underestimated, probably due to a poor adaptation of the virus grown in these cell lines to E-11. Based on all these results, E-11 seems to be the most appropriate cell for in vitro culture of RGNNV, SJNNV, and reassortant strains.

Immune Response of Senegalese Sole against Betanodavirus Mutants with Modified Virulence.

Nervous necrosis virus (NNV), genus Betanodavirus, the etiological agent of the viral encephalopathy and retinopathy (VER), presents a genome with two positive-sense single-stranded RNA segments. Striped jack nervous necrosis virus (SJNNV) and red-spotted grouper nervous necrosis virus (RGNNV), together with reassortants RGNNV/SJNNV, are the betanodaviruses predominantly isolated in Southern Europe. An RGNNV/SJNNV reassortant isolated from Senegalese sole (wt160) causes high mortalities in this fish species. This virus presents differences in the sequence of the 3' non-coding region (NCR) of both segments compared to RGNNV and SJNNV reference strains. Previously, it has been reported that the reversion of two of these differences (nucleotides 1408 and 1412) in the RNA2 3'NCR to the SJNNV-type (recombinant r1408-1412) resulted in a decrease in sole mortality. In the present study, we have applied an OpenArray® to analyse the involvement of sole immune response in the virulence of several recombinants: the r1408-1412 and two recombinants, developed in the present study, harbouring mutations at positions 3073 and 3093 of RNA1 3'NCR to revert them to RGNNV-type. According to the correlation values and to the number of expressed genes, the infection with the RNA2-mutant provoked the most different immune response compared to the immune response triggered after the infection with the rest of the viruses, and the exclusive and high upregulation of genes related to the complement system. The infection with the RNA1-mutants also provoked a decrease in mortality and their replication was delayed at least 24 h compared to the wt160 replication, which could provoke the lag observed in the immune response. Furthermore, the infection with the RNA1-mutants provoked the exclusive expression of pkr and the downregulation of il17rc.

Antiviral Activity of Carrageenans and Processing Implications.

Carrageenan and carrageenan oligosaccharides are red seaweed sulfated carbohydrates with well-known antiviral properties, mainly through the blocking of the viral attachment stage. They also exhibit other interesting biological properties and can be used to prepare different drug delivery systems for controlled administration. The most active forms are λ-, ι-, and κ-carrageenans, the degree and sulfation position being determined in their properties. They can be obtained from sustainable worldwide available resources and the influence of manufacturing on composition, structure, and antiviral properties should be considered. This review presents a survey of the antiviral properties of carrageenan in relation to the processing conditions, particularly those assisted by intensification technologies during the extraction stage, and discusses the possibility of further chemical modifications.

Psychometric properties of the European Portuguese version of the Wijma Delivery Expectancy/Experience Questionnaire in pregnant women.

The researchers' aim is to examine the psychometric properties of the European Portuguese version of the Wijma Delivery Expectancy/Experience Questionnaire version A (WDEQ-A). Data from 669 pregnant Portuguese women was collected between June and October of 2019. From the initial 33-item Portuguese version of the WDEQ-A, a 27-item instrument was developed. Five-factor solution explained 63.8% of the total variance. The factors defined by the researchers were: fear/lack of self-efficacy, loneliness, negative appraisal, lack of positive anticipation and concern for the child. Cronbach's alpha coefficient was 0.934. The European Portuguese version of the WDEQ-A is a reliable and valid tool to measure fear of childbirth on both nulliparous and multiparous women and can be a useful tool for cross-cultural research. Moreover, researchers support the use of this tool in clinical practice to recognize and address fearful women during prenatal period.

Development and Validation of a SYBR Green Real Time PCR Protocol for Detection and Quantification of Nervous Necrosis Virus (NNV) Using Different Standards.

The nervous necrosis virus (NNV) is a threat to fish aquaculture worldwide, especially in Mediterranean countries. Fast and accurate diagnosis is essential to control it, and viral quantification is required to predict the level of risk of new viral detections in field samples. For both, reverse transcription real-time quantitative polymerase chain reaction (RT-qPCR) is used by diagnostic laboratories. In the present study, we developed an RT-qPCR procedure for the diagnosis and simultaneous quantification of NNV isolates from any of the four genotypes. The method proved to be highly sensitive in terms of crude virus titer: 5.56-9.88 TCID50/mL (tissue culture infectious dose per mL), depending on the viral strain, and averaging 8.8 TCID50/mL or 0.08 TCID50/reaction. Other standards also yielded very low detection limits: 16.3 genome copies (cps) of purified virus per mL, 2.36 plasmid cps/mL, 7.86 in vitro synthetized RNA cps/mL, and 3.16 TCID50/mL of virus from infected tissues. The diagnostic parameters evaluated in fish samples were much higher in comparison to cell culture isolation and nested PCR. In addition, the high repeatability and reproducibility of the procedure, as well as the high coefficient of determination (R2) of all the calibration curves with any type of standard tested, ensure the high reliability of the quantification of NNV using this RT-qPCR procedure, regardless of the viral type detected and from the type of standard chosen.

Fear of childbirth in time of the new coronavirus pandemic.

OBJECTIVE: Reflect on how the new coronavirus pandemic triggered or accentuated the fear of childbirth in pregnant women and affected childbirth care practices. METHODS: Reflective analysis of women's pregnancy and childbirth experiences during the current pandemic, supported by the latest scientific evidence and recommendations on the topic. RESULTS: Pregnancy and childbirth are life-changing events for women, but during the new coronavirus pandemic, fear and uncertainty have taken on an unprecedented dimension in the negative way that many pregnant women have anticipated and experienced childbirth. FINAL CONSIDERATIONS: The current period has accentuated a chronic problem: a paternalistic system of health institutions in the approach to childbirth, dense with additional levels of fear in pregnant women. In this context, addressing the fear of childbirth means not giving up the promotion of safe and positive birth experiences for women.

Midwifery interventions to reduce fear of childbirth in pregnant women: a scoping review protocol.

OBJECTIVE: The objective of the review is to map and analyze midwifery interventions to reduce fear of childbirth in pregnant women. INTRODUCTION: Fear of childbirth is a phenomenon negatively affecting women's health and well-being before and during pregnancy, as well as after childbirth. During the previous few decades, there has been a growing interest in research into interventions to reduce the fear of childbirth in childbearing women. Currently, providing an appropriate model of care for pregnant women with fear of childbirth is a challenge in midwifery care. Therefore, further efforts are needed to identify and examine the characteristics of different midwifery interventions to reduce fear of childbirth in pregnant women. INCLUSION CRITERIA: This scoping review will consider studies that include interventions to reduce fear of childbirth in pregnant women, led and implemented by midwives, during the antenatal period, in all possible birth scenarios. Quantitative, qualitative, and mixed methods studies will be included. METHODS: The JBI methodology for conducting scoping reviews will be employed. Published and unpublished literature in English, Portuguese, and Spanish, from 1981 to the present, will be included. MEDLINE, CINAHL Complete, Scopus, Web of Science, Embase, and Cochrane Library databases will be searched. Searches for gray literature will be performed. Data will be extracted using a tool developed specifically for the scoping review objectives.

Differential immunogene expression profile of European sea bass (Dicentrarchus labrax, L.) in response to highly and low virulent NNV.

European sea bass is highly susceptible to the nervous necrosis virus, RGNNV genotype, whereas natural outbreaks caused by the SJNNV genotype have not been recorded. The onset and severity of an infectious disease depend on pathogen virulence factors and the host immune response. The importance of RGNNV capsid protein amino acids 247 and 270 as virulence factors has been previously demonstrated in European sea bass; however, sea bass immune response against nodaviruses with different levels of virulence has been poorly characterized. Knowing the differences between the immune response against both kinds of isolates may be key to get more insight into the host mechanisms responsible for NNV virulence. For this reason, this study analyses the transcription of immunogenes differentially expressed in European sea bass inoculated with nodaviruses with different virulence: a RGNNV virus obtained by reverse genetics (rDl956), highly virulent to sea bass, and a mutated virus (Mut247+270Dl956, RGNNV virus displaying SJNNV-type amino acids at positions 247 and 270 of the capsid protein), presenting lower virulence. This study has been performed in brain and head kidney, and the main differences between the immunogene responses triggered by both viruses have been observed in brain. The immunogene response in this organ is stronger after inoculation with the most virulent virus, and the main differences involved genes related with IFN I system, inflammatory response, cell-mediated response, and apoptosis. The lower virulence of Mut247+270Dl956 to European sea bass can be associated with a delayed IFN I response, as well as an early and transitory inflammation and cell-mediated responses, suggesting that those can be pivotal elements in controlling the viral infection, and therefore, their functional activity could be analysed in future studies. In addition, this study supports the role of capsid amino acids at positions 247 and 270 as important determinants of RGNNV virulence to European sea bass.

Betanodavirus and VER Disease: A 30-year Research Review.

The outbreaks of viral encephalopathy and retinopathy (VER), caused by nervous necrosis virus (NNV), represent one of the main infectious threats for marine aquaculture worldwide. Since the first description of the disease at the end of the 1980s, a considerable amount of research has gone into understanding the mechanisms involved in fish infection, developing reliable diagnostic methods, and control measures, and several comprehensive reviews have been published to date. This review focuses on host-virus interaction and epidemiological aspects, comprising viral distribution and transmission as well as the continuously increasing host range (177 susceptible marine species and epizootic outbreaks reported in 62 of them), with special emphasis on genotypes and the effect of global warming on NNV infection, but also including the latest findings in the NNV life cycle and virulence as well as diagnostic methods and VER disease control.

Fear of childbirth in time of the new coronavirus pandemic / El temor al parto en tiempos de pandemia del nuevo coronavirus / O medo do parto em tempo de pandemia do novo coronavírus

ABSTRACT Objective: Reflect on how the new coronavirus pandemic triggered or accentuated the fear of childbirth in pregnant women and affected childbirth care practices. Methods: Reflective analysis of women's pregnancy and childbirth experiences during the current pandemic, supported by the latest scientific evidence and recommendations on the topic. Results: Pregnancy and childbirth are life-changing events for women, but during the new coronavirus pandemic, fear and uncertainty have taken on an unprecedented dimension in the negative way that many pregnant women have anticipated and experienced childbirth. Final considerations: The current period has accentuated a chronic problem: a paternalistic system of health institutions in the approach to childbirth, dense with additional levels of fear in pregnant women. In this context, addressing the fear of childbirth means not giving up the promotion of safe and positive birth experiences for women.

RESUMEN Objetivo: Reflejar acerca de cómo la pandemia de COVID-19 desencadenó o acentuó el temor al parto en las embarazadas y afectó las prácticas de asistencia al parto. Métodos: Análisis reflexivo sobre las experiencias de embarazo y parto de mujeres, durante la actual pandemia, bajo el soporte de las más recientes evidencias científicas y recomendaciones sobre el tema. Resultados: El embarazo y el parto son acontecimientos transformadores en la vida de las mujeres, pero, en el curso del nuevo coronavirus, el temor y la incertidumbre ganaron una dimensión sin precedentes negativamente como muchas embarazadas han anticipado y experimentado el parto. Consideraciones finales: Actualmente acentuó un problema crónico: un sistema paternalista de las instituciones de salud en el abordaje al parto, adensado por niveles adicionales de temor en las embarazadas. Así, abordar el temor al parto significa no abdicar de la promoción de experiencias seguras y positivas del parto para las mujeres.

RESUMO Objetivo: Refletir acerca da forma como a pandemia do novo coronavírus desencadeou ou acentuou o medo do parto nas gestantes e afetou as práticas de assistência ao parto. Métodos: Análise reflexiva sobre as experiências de gravidez e parto de mulheres, durante a atual pandemia, sob o suporte das mais recentes evidências científicas e recomendações sobre o tema. Resultados: A gravidez e o parto são acontecimentos transformadores na vida das mulheres, mas, no decorrer da pandemia do novo coronavírus, o medo e a incerteza ganharam uma dimensão sem precedentes na forma negativa como muitas gestantes têm antecipado e experienciado o parto. Considerações finais: O período atual acentuou um problema crónico: um sistema paternalista das instituições de saúde na abordagem ao parto, adensado por níveis adicionais de medo nas gestantes. Nesse contexto, abordar o medo do parto significa não abdicar da promoção de experiências seguras e positivas de parto para as mulheres.

Capsid amino acids at positions 247 and 270 are involved in the virulence of betanodaviruses to European sea bass.

European sea bass (Dicentrarchus labrax) is severely affected by nervous necrosis disease, caused by nervous necrosis virus (NNV). Two out of the four genotypes of this virus (red-spotted grouper nervous necrosis virus, RGNNV; and striped jack nervous necrosis virus, SJNNV) have been detected in sea bass, although showing different levels of virulence to this fish species. Thus, sea bass is highly susceptible to RGNNV, whereas outbreaks caused by SJNNV have not been reported in this fish species. The role of the capsid protein (Cp) amino acids 247 and 270 in the virulence of a RGNNV isolate to sea bass has been evaluated by the generation of recombinant RGNNV viruses harbouring SJNNV-type amino acids in the above mentioned positions (Mut247Dl965, Mut270Dl965 and Mut247 + 270Dl965). Viral in vitro and in vivo replication, virus virulence and fish immune response triggered by these viruses have been analysed. Mutated viruses replicated on E-11 cells, although showing some differences compared to the wild type virus, suggesting that the mutations can affect the viral cell recognition and entry. In vivo, fish mortality caused by mutated viruses was 75% lower, and viral replication in sea bass brain was altered compared to non-mutated virus. Regarding sea bass immune response, mutated viruses triggered a lower induction of IFN I system and inflammatory response-related genes. Furthermore, mutations caused changes in viral serological properties (especially the mutation in amino acid 270), inducing higher seroconversion and changing antigen recognition.