The micro‐neutralization assay is a fundamental test in virology, immunology, vaccine assessment, and epidemiology studies. Since the SARS‐CoV‐2 outbreak at the end of December 2019 in China, it has become extremely important to have well‐established and validated diagnostic and serological assays for this new emerging virus. Here, we present a micro‐neutralization assay with the use of SARS‐CoV‐2 wild type virus with two different methods of read‐out. We evaluated the performance of this assay using human serum samples taken from an Italian seroepidemiological study being performed at the University of Siena, along with the human monoclonal antibody CR3022 and some iper‐immune animal serum samples against Influenza and Adenovirus strains. The same panel of human samples have been previously tested in enzyme‐linked immunosorbent assay (ELISA) as a pre‐screening. Positive, borderline, and negative ELISA samples were evaluated in neutralization assay using two different methods of read‐out: subjective (by means of an inverted optical microscope) and objective (by means of a spectrophotometer). Our findings suggest that at least 50% of positive ELISA samples are positive in neutralization as well, and that method is able to quantify different antibody concentrations in a specific manner. Taken together, our results confirm that the colorimetric cytopathic effect‐based microneutralization assay could be used as a valid clinical test method for epidemiological and vaccine studies.

Influenza is a vaccine preventable disease and vaccination remains the most effective method of controlling the morbidity and mortality of seasonal influenza, especially with respect to risk groups. To date, three types of influenza vaccines have been licensed: inactivated, live-attenuated, and recombinant haemagglutinin vaccines. Effectiveness studies allow an assessment of the positive effects of influenza vaccines in the field. The effectiveness of current influenza is suboptimal, being estimated as 40% to 60% when the vaccines strains are antigenically well-matched with the circulating viruses. This review focuses on influenza viruses and vaccines and the role of vaccine effectiveness studies for evaluating the benefits of influenza vaccines. Overall, influenza vaccines are effective against morbidity and mortality in all age and risk groups, especially in young children and older adults. However, the effectiveness is dependent on several factors such as the age of vaccinees, the match between the strain included in the vaccine composition and the circulating virus, egg-adaptations occurring during the production process, and the subject’s history of previous vaccination.

Influenza D virus is a novel influenza virus, which was first isolated from an ailing swine in 2011 and later detected in cattle, suggesting that these animals may be a primary natural reservoir. To date, few studies have been performed on human samples and there is no conclusive evidence on the ability of the virus to infect humans. The aim of this serological study was to assess the prevalence of antibodies against influenza D virus in human serum samples collected in Italy from 2005 to 2017. Serum samples were analysed by haemagglutination inhibition and virus neutralization assays. The results showed that the prevalence of antibodies against the virus increased in the human population in Italy from 2005 to 2017, with a trend characterized by a sharp increase in some years, followed by a decline in subsequent years. The virus showed the ability to infect and elicit an immune response in humans. However, prevalence peaks in humans appear to follow epidemics in animals and not to persist in the human population.

Toxoplasmosis is a worldwide health problem. Infection in pregnant women can result in severe fetal morbidity or in subclinical neonatal infection; most subclinical cases develop ocular and neurological sequelae. The purpose of this serological study was to assess the prevalence of Toxoplasma gondii in two populations of women of childbearing age in Siena (Tuscany, Central Italy) and Bari (Apulia, Southern Italy) between 2013 and 2017 and in a group of pregnant women in Bari in 2016–2017. Serum samples were tested for the presence of specific anti-Toxoplasma gondii IgG antibodies by a commercially available ELISA test. The percentage of seropositive subjects in Bari was significantly higher than in Siena (22.4% vs. 12.4%) and an age-related trend was observed. A low prevalence of T. gondii infection (13.8%) was observed among the pregnant women tested. In addition to showing a significant difference between Central and Southern Italy, this study provides updated data on T. gondii seroprevalence in women during childbearing age and pregnancy. The results confirm a trend toward a decrease, especially in younger people and pregnant women.

Introduction: Current influenza vaccines are mainly produced from viruses propagated in eggs, an established process that has been developed over 70 years. However, this technology presents some drawbacks and other platforms are under development or have been already developed in order to replace or to be used alongside the old one. Area covered: The present review provides an overview of influenza vaccine production, starting from egg-based technology, to cell-derived vaccines, until the novel platforms and technologies for the production of influenza vaccines such as DNA-based vaccines, virus-like particles and plant-based technology. Expert opinion: The ideal method of production should have certain characteristics such as great flexibility and scalability, the viruses should be representative of the circulating strains, process should be standardized and controlled, and it should be possible to start production as soon as the sequence of the new influenza strain is available. However, it is important not to underestimate the fact that some parts of the vaccine production process have been established for egg-based vaccines. Thus, changes in vaccine production methods are not merely “technical changes”; rather, they involve various aspects that slow down the authorization of new influenza vaccines and make the complete replacement of egg-based production unlikely in the near future.

Newly emerging or re-emerging infections continue to pose significant global public health threats. This chapter provides an overview of the combinations of factors that led to the emergence of arthropod-borne viruses as human and veterinary health threats, in order to understand the risk associated and how this can be mitigated. Considering the history of emergence of some arboviruses, these epidemics have occurred globally as a result of climate and socioeconomic changes that have allowed the spread to new geographical areas of viruses previously confined to specific ecological niches such as West Nile and Chikungunya, or viruses considered under control such as Dengue, Japanese encephalitis, and Yellow fever. Moreover, the greatest risk for humans derives from the ability of these viruses to adopt transmission cycles involving highly anthropophilic mosquito species. Finally, many other arboviruses are largely ignored despite their potential to emerge globally. The recent epidemic spread of Zika virus throughout the Americas is the evidence that arboviruses are likely to continually emerge and re-emerge and that improved scientific technologies and knowledge is essential to deal with future vector-borne epidemics. Research priorities should therefore focus on surveillance systems and vector control tools, as well as on the development of antiviral molecules or candidate vaccine.