Giovanni Anselmi

A New Chance of Preventing Winter Diseases by the Administration of Synbiotic Formulations

Background The efficacy of probiotics is currently well documented with regard to the improvement of gastrointestinal functions, whereas their potential role in the prevention of infectious respiratory diseases has not been sufficiently analyzed. Purpose of the Study and Methods A 3-stage prospective, randomized, double blind, placebo-controlled study was carried out with several synbiotic preparations containing 3 to 5 strains of Lactobacillus plantarum, Lactobacillus rhamnosus, and Bifidobacterium lactis, lactoferrin and prebiotics such as either FOS (short-chain fructooligosaccharides) or GOS (galactooligosaccharides). The study was performed over 3 different winter seasons between 2003 and 2007, and was aimed at assessing the ability of the different preparations to improve intestinal functions and to increase the bodys defences against respiratory infections. In 2003/04 (stage 1; 237 healthy volunteers) an active formulation (A) containing 3 probiotic strains and FOS was used versus placebo; in 2005/06 (stage 2; 234 healthy volunteers) the same formulation versus a similar preparation enriched with lactoferrin (B), and versus placebo; in 2006/07 (stage 3; 250 healthy volunteers), 2 new synbiotic formulations each containing 5 probiotics and FOS (C) or GOS (D), respectively, versus placebo. Results In stage 1, bowel functions improved (P=0.004) in terms of reduced bloating and more regular intestinal motility. The length of acute respiratory infection episodes considered as a whole (−0.97 d; P=0.007) and upper respiratory tract infections (URTIs, −1.96 d; P=0.044) were significantly decreased in the synbiotic group. The severity of episodes recorded a statistically significant drop in both episodes considered as a whole (3.21 average score vs. 3.98 in the placebo group, P<0.001) and in URTI (2.56 vs. 3.82; P=0.004) and flu classes (3.80 vs. 4.67, P=0.001). In stage 2, improvement of bowel functions was statistically significant (P=0.005) in synbiotic preparation A. A statistically significant reduction in the number of respiratory tract infections episodes was noted with both the two active formulations (P=0.002 in group A and P=0.003 in group B). The duration of episodes considered as a whole (−1.12 d in one of the 2 active formulation groups; P=0.005), URTIs (−2.08 d in group A; P=0.036) and influenza-like illness episodes (−1.40 d in group A; P=0.049) was significantly decreased in the synbiotic group. A reduction trend in cold episodes was also recorded. The severity of episodes recorded a statistically significant drop in episodes considered as a whole (−0.73 in group A, P=0.003; −0.65 in group B, P=0.004) and in the case of flu (−1.25 in group A, P<0.001; −1.18 in group B, P<0.001). In stage 3, the improvement of bowel functions was confirmed for both active formulations (P<0.001). A significant decrease in the total length of respiratory episodes (−1.51 d; P<0.001 in the group C and −1.39 d; P<0.001 in the group D) and in the length of cough (−3.08 d; P<0.001 in group C; −2.83 d; P<0.001 in group D), cold (−1.02 d; P=0.019 in group C; −1.32 d; P=0.001 in group D) and flu episodes was reported. The severity of episodes recorded a statistically significant drop in regard to episodes considered as a whole, the cold and flu classes in both groups and for cough too in group C. The number of episodes also dropped considerably in terms of overall episodes, cold (group C) and flu. Conclusions These results demonstrate that a regular, long-term intake of various synbiotics may improve health by reducing the incidence and severity of respiratory diseases during the cold season.

Comparison of RT‐PCR with other diagnostic assays for rapid detection of influenza viruses

To compare the effectiveness of reverse transcription‐polymerase chain reaction (RT‐PCR), shell vial culture and cytospin assay as laboratory techniques for rapid diagnosis of influenza infections, a retrospective study was carried out on 270 aliquots of oropharyngeal swabs collected from October 1993 to March 1996 and already characterized by standard isolation procedures, and a prospective study in which 65 clinical samples taken from patients with influenza‐like syndrome between October 1996 and March 1997 were tested. In the retrospective study, using conventional isolation as the gold standard, the sensitivity of RT‐PCR and cytospin assay for virus A was 100% (95% confidence interval (CI), 89.1–100) and for virus B it was 100% (95% CI, 56.1–100) compared with 77.5% (95% CI, 61.1–88.6) and 71.4% (95% CI, 30.3–94.9) for shell vial culture. The specificity of all the three assays was 100% (95% CI, 98.0–100) for virus A and 100% (95% CI, 98.2–100) for virus B. In the prospective study the sensitivity of RT‐PCR was greater than that of the other tests considered, both rapid and standard. It is suggested that RT‐PCR should be employed in combination with conventional culture techniques in routine diagnosis of influenza infections in order to obtain results more rapidly and to improve virus detection even in circumstances in which standard isolation could be problematic. J. Med. Virol. 56: 168–173, 1998.

Response to 2009 pandemic and seasonal influenza vaccines co-administered to HIV-infected and HIV-uninfected former drug users living in a rehabilitation community in Italy.

BACKGROUND 2009 A(H1N1) pandemic influenza vaccination was recommended as a priority to essential workers and high-risk individuals, including HIV-infected patients and people living in communities. METHODS HIV-infected and HIV-uninfected former drug-users (18-60 years old) living in a rehabilitation community (San Patrignano, Italy) received one dose of a MF59-adjuvanted 2009 pandemic influenza vaccine and one dose of a 2009-2010 seasonal trivalent inactivated influenza vaccine (containing A/Brisbane/59/2007(H1N1), A/Brisbane/10/2007(H3N2), B/Brisbane/60/2008) simultaneously. Antibodies against each vaccine antigen were determined at the time of vaccination and one and six months post-vaccination by hemagglutination-inhibition test. RESULTS 49 HIV-infected and 60 HIV-uninfected subjects completed the study. Most (98%) HIV-infected participants were on antiretroviral treatment, the median CD4+ cell count was 350 (IQR 300)cells/μl and viremia was suppressed in 91.8% of cases. One month post-vaccination, no significant changes in immune-virological parameters were observed. One month post-vaccination, the immune responses to both pandemic and seasonal vaccine met the EMA-CPMP criteria for immunogenicity of influenza vaccines in both HIV-infected and HIV-uninfected subjects. No difference in vaccine responses was observed between the two groups. Six months after vaccination, the percentages of vaccinees with antibody titres ≥1:40 and antibody geometric mean titres significantly decreased in both groups. However, they were significantly lower in HIV-infected than in HIV-uninfected vaccinees. In subjects who had been primed to seasonal influenza the year before (through either vaccination or natural infection), levels of antibodies against 2009 A(H1N1) were higher than those measured in unprimed subjects, both one month and six months post-vaccination. CONCLUSIONS The co-administration of a single dose of 2009 pandemic MF59-adjuvanted influenza vaccine with a seasonal vaccine provided a protective immune response in both HIV-infected and HIV-uninfected individuals. Subjects who had been primed to seasonal influenza in the year preceding the pandemic had a more vigorous and long-lasting antibody response to 2009 pandemic vaccine.

Genetic drift influenza A(H3N2) virus hemagglutinin (HA) variants originated during the last pandemic turn out to be predominant in the 2011–2012 season in Northern Italy

Influenza A(H3N2) virus is once again the predominant strain after the 2009 pandemic. Its molecular epidemiology and phylogeny were investigated during the 2011-2012 season in Northern Italy. The epidemiological and virological influenza surveillance was carried out within the framework of the Italian Influenza Surveillance Network. The hemagglutinin (HA) gene of the A(H3N2) viruses detected was analyzed by means of a time-scaled phylogenetic approach. In Northern Italy, the 2011-2012 epidemic wave was sustained almost exclusively by influenza A(H3N2) viruses (87.2% of total influenza virus detections). The consultation rates for influenza-like illness (ILI) in the age group ≥65 years were 1.5 to 6-fold higher than those registered during the previous eight epidemics: A(H3N2) was the only virus identified in this group. The phylogenetic analysis of A(H3N2) viruses showed viruses belonging to the A/Victoria/208/2009 genetic clade, characterized by substitutions in HA antigenic sites with respect to the A/Perth/16/2009-like 2011-2012 vaccine strain. About one-third of analyzed sequences fell into group 6 and two thirds into group 3 (subdivided into 3A, 3B, and 3C). The time scale reconstruction of the phylogeny showed several independent introductions of A(H3N2) groups between summer and winter of 2011. However, the common origin of all the circulating A(H3N2) strains dated back to the 2009 pandemic period (November 2009). The time scale phylogenetic approach is of particular importance for the evaluation of the introduction and circulation of new variants in the area. Therefore, it should be implemented within the framework of influenza virological surveillance.

Ten years (2004-2014) of influenza surveillance in Northern Italy

As the regional influenza reference centre operating within the Italian network InfluNet, here we report data on virological and epidemiological surveillance of influenza, as well as on the vaccination coverage rates achieved in Lombardy (Northern Italy) over 10 consecutive winter seasons (2004–2014). Over the past 10 years, influenza vaccine coverage declined both in the general population (from 15.7% in 2004–2005 to 11.7% in 2013–2014) and in the vaccine-target population of individuals ≥65-y-of-age (from 65.3% in 2004–2005 to 48.6% in 2013–2014) and is far below the minimum planned threshold level (75%). The highest influenza-like illness (ILI) rates were recorded during the 2004–2005 and 2009–2010 epidemics (peak incidence: 12.04‰ and 13.28‰, respectively). Both seasons were characterised by the introduction of novel viral strains: A/Fujian/411/2002(H3N2) (a drifted hemagglutinin variant) and A/California/7/2009(H1N1) pandemic virus (a swine origin quadruple reassortant), respectively. Because the antigenic match between vaccine and circulating strains was good in both of these seasons, a relevant proportion of cases may have been prevented by vaccination. A different situation was observed during the 2011–2012 season, when ILI morbidity rates in individuals ≥65-y-of-age were 1.5–6-fold higher than those registered during the other epidemics under review. The higher morbidity resulted from the circulation during the 2011–2012 season of an A/Victoria/361/2011(H3N2)-like variant that presented a reduced genetic match with the A(H3N2) strain included in the 2011–2012 vaccine composition. The continuous surveillance of the characteristics of circulating viruses is an essential tool for monitoring their matching with seasonal vaccine strains. Strategies to increase coverage rates are warranted.

Molecular characterization of influenza viruses circulating in Northern Italy during two seasons (2005/2006 and 2006/2007) of low influenza activity.

The influenza activity and circulation of influenza viruses in Lombardy (the most populous Italian region) were observed during two consecutive seasons (2005/2006 and 2006/2007) characterized by low influenza activity by the Italian Influenza Surveillance Network. The molecular characteristics of circulating viruses were analyzed to evaluate the introduction of new variants and emergence of vaccine‐escape viruses. In both seasons, the epidemic in Lombardy was sustained almost exclusively by influenza A viruses, accounting for 80.5% and 93.6% of total detections, respectively, and the co‐circulation of A/H3 viruses belonging to distinct phylogenetic groups was observed. The A/H1N1 viruses isolated during the 2005/2006 season were closely related to A/New Caledonia/20/99, while the hemagglutinin (HA) sequences of the A/H1N1 viruses from the 2006/2007 season exhibited a greater diversity. These viruses were A/Solomon Islands/3/2006‐like and showed several variants. All B isolates were similar to B/Malaysia/2506/2004 belonging to the B/Victoria/2/87‐lineage. Influenza B virus was the dominant virus in Europe in the 2005/2006 season and accounted for the 20% of total detections in Lombardy. Overall, the viruses studied presented heterogeneity in their HA sequences suggesting the circulation of a miscellaneous set of variants during the two seasons notwithstanding the medium‐low activity of influenza. The importance of virological surveillance of influenza viruses is recognized widely and the molecular characterization of the viruses, especially in vaccinated subjects, is of particular importance to evaluate the introduction and circulation of new variants. J. Med. Virol. 80:1984–1991, 2008.

Long-term patterns of humoral and cellular response after vaccination against influenza A (H1N1) in patients with hematologic malignancies

The efficacy of a novel vaccine against influenza virus A (H1N1) in patients with hematologic malignancies is largely unknown.

Surveillance of influenza viruses in the post-pandemic era (2010-2012) in Northern Italy.

The activity and circulation of influenza viruses in Lombardy — Northern Italy — (a region with nearly 10 out of the 60 million inhabitants of Italy) were investigated during two consecutive seasons (2010–2011 and 2011–2012), as part of the Italian Influenza Surveillance Network. The molecular characteristics of the hemagglutinin (HA) sequence of circulating viruses were analyzed to investigate the emergence of influenza viral variants. In the surveyed area, the influenza activity of these two post-pandemic seasons was similar in terms of both time frame and impact. The timing of the influenza epidemics was similar to the timing seen prior to the emergence of the pandemic A(H1N1) virus in 2009. A(H1N1)pdm09 was the predominant virus circulating during the 2010–2011 post-pandemic season and then—unexpectedly—almost disappeared. The HA sequences of these A(H1N1)pdm09 viruses segregated in a different genetic group with respect to those identified during the 2009 pandemic, although they were still closely related to the vaccine viral strain A/California/07/2009. Influenza A(H3N2) viruses were the predominant viruses circulating during the 2011–2012 season, accounting for nearly 88% of influenza viruses identified. All HA sequences of the A(H3N2) viruses isolated in the 2011–2012 season fell into the A/Victoria/208/2009 genetic clade (although the A/Perth/16/2009 virus was the reference vaccine strain). B viruses presented with a mixed circulation of viral variants during these two seasons: viruses belonging to both B/Victoria and B/Yamagata lineages co-circulated in different proportions, with a notable rise in the proportion of B/Yamagata viruses (B/Wisconsin/1/2010-like) during the 2011–2012 epidemic. In conclusion, the continuous monitoring of the characteristics of circulating viruses is an essential tool for understanding the epidemiological and virological features of influenza viruses, for monitoring their matching with seasonal vaccine strains, and for tuning vaccination strategies.

Italian influenza surveillance network: Results of the first year of activity

The spread of influenza is of major public health concern. Community based clinical morbidity data constitute an excellent predictor of when the peak of influenza virus activity will be reached. Surveillance programmes exist in many countries [1, 2]. A voluntary system for the surveillance of acute respiratory infection (ARI) and influenza was established in Italy in 1996. The surveillance is based on a sentinel network of general practitioners (GPs) and pediatricians (Ps) for the collection of clinical data and a network of virological laboratories. The physicians report to the Istituto di Virologia on a weekly basis.

Epidemiology and molecular characterization of influenza viruses, human parechoviruses and enteroviruses in children up to 5 years with influenza-like illness in Northern Italy during seven consecutive winter seasons (2010–2017)

Besides the influenza virus (IV), several other viruses are responsible for influenza-like illness (ILI). Although human parechoviruses (HPeVs) and enteroviruses (EVs) may impact on ILI, limited data on their epidemiological characteristics are available. During seven consecutive winter seasons (from 2010-2011 to 2016-2017), within the framework of an influenza surveillance system (InfluNet), 593 respiratory swabs were collected from children ≤5 years of age with ILIs. Molecular detection showed that 58.3 % of swabs were positive for at least one of the viruses under study: 46 % for IV, 13 % for EV and 5.4 % for HPeV. A single virus was identified in 51.3 % of samples while more than one virus was detected in 7 % of the samples. The risk of contracting IV was higher than the risk associated with EV, which in turn was higher than the risk of contracting HPeV. The risk of developing an IV infection was twofold greater in children >3 years than in those ≤3 years, who had higher risk of EV/HPeV infection. The frequency of EV/HPeV-positive swabs increased significantly during the 2016-2017 winter season compared to the previous six seasons. Sixteen EV genotypes were identified belonging to species A and B. HPeV-1 was the most frequently detected genotype, followed by -6 and -3. In this study, IV was mainly responsible for ILI, however EV and HPeV were also involved and particularly affected children ≤3 years of age. Influenza surveillance samples could provide us with valuable insight into the epidemiological features of viruses involved in ILI.