Caterina Vocale

West Nile virus in Europe: emergence, epidemiology, diagnosis, treatment, and prevention

West Nile virus (WNV), a mosquito-borne flavivirus in the Japanese encephalitis antigenic group, has caused sporadic outbreaks in humans, horses and birds throughout many of the warmer regions of Europe for at least 20 years. Occasional cases of West Nile encephalitis have also been associated with infected blood transfusions and organ donations. Currently, WNV appears to be expanding its geographical range in Europe and causing increasing numbers of epidemics/outbreaks associated with human morbidity and mortality. This brief review reports on the current epidemic situation regarding WNV in Europe, highlighting the clinical, diagnostic and preventive measures available for controlling this apparently emerging human pathogen.

Human infection with highly pathogenic A(H7N7) avian influenza virus, Italy, 2013.

During an influenza A(H7N7) virus outbreak among poultry in Italy during August–September 2013, infection with a highly pathogenic A(H7N7) avian influenza virus was diagnosed for 3 poultry workers with conjunctivitis. Genetic analyses revealed that the viruses from the humans were closely related to those from chickens on affected farms.

Diagnosis of West Nile Virus Human Infections: Overview and Proposal of Diagnostic Protocols Considering the Results of External Quality Assessment Studies

West Nile virus, genus Flavivirus, is transmitted between birds and occasionally other animals by ornithophilic mosquitoes. This virus also infects humans causing asymptomatic infections in about 85% of cases and <1% of clinical cases progress to severe neuroinvasive disease. The virus also presents a threat since most infections remain unapparent. However, the virus contained in blood and organs from asymptomatically infected donors can be transmitted to recipients of these infectious tissues. This paper reviews the presently available methods to achieve the laboratory diagnosis of West Nile virus infections in humans, discussing the most prominent advantages and disadvantages of each in light of the results obtained during four different External Quality Assessment studies carried out by the European Network for ‘Imported’ Viral Diseases (ENIVD).

The experience of West Nile virus integrated surveillance system in the Emilia-Romagna region: five years of implementation, Italy, 2009 to 2013

Predicting West Nile virus (WNV) circulation and the risk of WNV epidemics is difficult due to complex interactions of multiple factors involved. Surveillance systems that timely detect virus activity in targeted areas, and allow evidence-based risk assessments may therefore be necessary. Since 2009, a system integrating environmental (mosquitoes and birds) and human surveillance has been implemented and progressively improved in the Emilia-Romagna region, Italy. The objective is to increase knowledge of WNV circulation and to reduce the probability of virus transmission via blood, tissue and organ donation. As of 2013, the system has shown highly satisfactory results in terms of early detection capacity (the environmental surveillance component allowed detection of WNV circulation 3–4 weeks before human cases of West Nile neuroinvasive disease (WNND) occurred), sensitivity (capacity to detect virus circulation even at the enzootic level) and area specificity (capacity to indicate the spatial distribution of the risk for WNND). Strong correlations were observed between the vector index values and the number of human WNND cases registered at the province level. Taking into consideration two scenarios of surveillance, the first with environmental surveillance and the second without, the total costs for the period from 2009 to 2013 were reduced when environmental surveillance was considered (EUR 2.093 million for the first scenario vs EUR 2.560 million for the second). Environmental surveillance helped to reduce costs by enabling a more targeted blood unit testing strategy. The inclusion of environmental surveillance also increased the efficiency of detecting infected blood units and further allowed evidence-based adoption of preventative public health measures.

Toscana virus infections in northern Italy: laboratory and clinical evaluation.

Toscana virus (TOSv) is a neurotropic arthropod-borne virus that causes meningitis in the Mediterranean basin during the summer months. A total of 120 patients suffering from acute aseptic meningitis between July 1 and October 31, 2010 in northern Italy were evaluated. Eighteen of them (15%) were in the acute stage of TOSv disease.

Peri-Implant Response and Microflora in Organ Transplant Patients 1 Year after Prosthetic Loading: A Prospective Controlled Study.

BACKGROUND A recent study conducted in humans demonstrated for the first time that bone loss in the immediate period after implant insertion before loading did not significantly differ in organ transplant recipients with respect to normal subjects. PURPOSE The purpose of this study is to evaluate bone and periodontal response and peri-implant microflora in a group of organ-transplanted patients 1 year after prosthetic loading. MATERIALS AND METHODS The study population included 13 consecutive organ-transplanted (11 hearts, two livers) patients and 13 normal (healthy) control subjects who received 29 and 28 submerged dental implants, respectively. Crestal bone level, peri-implant probing depth (PIPD), and bleeding on probing were evaluated at prosthetic loading and after 1 year. Samples for microbiological testing were taken from the subgingival microbiota of each implant 1 year after loading and analyzed with polymerase chain reaction. All samples were examined for the presence of five microorganisms: Treponema denticola, Porphyromonas gingivalis, Prevotella intermedia, Tannarella forsythensis, and Actinobacillus actinomycetemcomitans. RESULTS A mean bone loss of 0.17 ± 0.10 and 0.20 ± 0.10 mm at 1 year was observed in the group of transplant recipients and in controls, respectively (N.S.). Similar results were obtained considering PIPD changes at 1 year (respectively 0.06 ± 0.71 mm in transplants vs 0.11 ± 0.74 mm in controls). Detection frequencies of pathogens were not statistically different between normal and transplanted patients. CONCLUSIONS The present pilot study seems to indicate that bone and periodontal response and microbiological status around submerged dental implants in immunocompromised organ-transplanted patients do not differ 1 year after loading from those observed in control patients and that this particular population of patients may be successfully rehabilitated with dental implants.

Human and entomological surveillance of Toscana virus in the Emilia-Romagna region, Italy, 2010 to 2012

Toscana virus (TOSV), transmitted by phlebotomine sandflies, is recognised as one of the most important causes of viral meningitis in summer in Mediterranean countries. A surveillance plan based on both human and entomological surveys was started in 2010 in the Emilia-Romagna region, Italy. Clinical samples from patients with neurological manifestations were collected during 2010 to 2012. The surveillance protocol was improved during these years, allowing the detection of 65 human infections. Most of these infections were recorded in hilly areas, where sandflies reach the highest density. Entomological sampling around the homes of the patients resulted in a low number of captured sandflies, while later sampling in a hilly area with high number of human cases (n=21) resulted in a larger number of captured sandflies. Using this approach, 25,653 sandflies were sampled, of which there were 21,157 females, which were sorted into 287 pools. TOSV RNA was detected by real-time PCR in 33 of the pools. The results highlighted the role of Phlebotomus perfiliewi as the main vector of TOSV and a potential link between vector density and virus circulation. This integrated system shows that an interdisciplinary approach improves the sensitiveness and effectiveness of health surveillance.

Comparison of Zika virus (ZIKV) RNA detection in plasma, whole blood and urine – Case series of travel-associated ZIKV infection imported to Italy, 2016

OBJECTIVES The capability to detect ZIKV RNA is of crucial importance for cases confirmation. However, due to the short-lived viremia, the detection of ZIKV RNA in plasma/serum is challenging for samples collected more than one week after onset of clinical illness. We compared the window time and detection rate of ZIKV RNA in different specimen types (plasma, whole blood and urine) collected simultaneously at several times post-symptom onset. METHODS We examined the presence of ZIKV RNA in matched specimens of whole blood, plasma and urine collected in the same date (3-28 days after symptom onset) from 10 ZIKV infected patients. RESULTS ZIKV RNA was found in plasma as late as 10 days after symptoms onset and tested positive in all 5 (100%) and in 2 of 6 (33,3%) plasma samples collected 1-5 and 6-10 days after symptoms onset, respectively. ZIKV RNA was positive in urine through the 21st day after symptom onset; the detection rate of ZIKV RNA in urine samples was 100% (11/11) for samples collected 1-10 days from symptoms onset, decreasing at later times of sampling. The detection rate of ZIKV RNA in whole blood was comparable to that in urine samples but extended the window of detection of ZIKV RNA up to 26 days after symptom onset. CONCLUSIONS Our results highlight the usefulness of simultaneously testing multiple specimen types in order to extend the rate and the time frame of ZIKV RNA detection, increasing the possibility of cases confirmation through direct diagnosis in convalescence-phase of infection, supplementing serological data which are often difficult to interpret.

Treponema denticola alters cell vitality and induces HO-1 and Hsp70 expression in porcine aortic endothelial cells

Treponema denticola is an oral spirochete that is associated with periodontal disease and detected occasionally in extraoral lesions associated with systemic disorders such as cardiovascular diseases. The effect of specific bacterial products from oral treponemes on endothelium is poorly investigated. This study analyzed the ability of components of the outer membrane of T. denticola (OMT) to induce apoptosis and heat shock proteins (HO-1 and Hsp70) in porcine aortic endothelial cells (pAECs), compared with results obtained with classical pro-inflammatory lipopolysaccharide (LPS) treatment. Cellular apoptosis was detected when pAECs were treated with either OMT or LPS, suggesting that OMT can damage endothelium integrity by reducing endothelial cell vitality. Stimulation with OMT, similarly to LPS response, increased HO-1 and Hsp-70 protein expression in a time-dependent manner, correlating with a rise in HO-1 and Hsp-70 mRNA. Collectively, these results support the hypothesis that T. denticola alters endothelial cell function. Moreover, our in vitro experiments represent a preliminary investigation to further in vivo study using a pig model to elucidate how T. denticola leaves the initial endodontic site and participates in the development of several systemic diseases.

Serological and molecular tools to diagnose visceral leishmaniasis: 2-years’ experience of a single center in Northern Italy

The diagnosis of visceral leishmaniasis (VL) remains challenging, due to the limited sensitivity of microscopy, the poor performance of serological methods in immunocompromised patients and the lack of standardization of molecular tests. The aim of this study was to implement a combined diagnostic workflow by integrating serological and molecular tests with standardized clinical criteria. Between July 2013 and June 2015, the proposed workflow was applied to specimens obtained from 94 in-patients with clinical suspicion of VL in the Emilia-Romagna region, Northern Italy. Serological tests and molecular techniques were employed. Twenty-one adult patients (22%) had a confirmed diagnosis of VL by clinical criteria, serology and/or real-time polymerase chain reaction; 4 of these patients were HIV-positive. Molecular tests exhibited higher sensitivity than serological tests for the diagnosis of VL. In our experience, the rK39 immunochromatographic test was insufficiently sensitive for use as a screening test for the diagnosis of VL caused by L. infantum in Italy. However, as molecular tests are yet not standardized, further studies are required to identify an optimal screening test for Mediterranean VL.