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Poster EJP Annual Meeting 2022

Il virus dell’epatite E (HEV) è un virus a RNA che causa, nell’uomo, un’epatite acuta generalmente autolimitante. I suini (domestici e selvatici) sono il principale reservoir. L’identifi cazione delle aziende infette è il presupposto principale per implementare dei programmi di sorveglianza per questa zoonosi, ormai considerata emergente, e per ridurre il rischio di introdurre prodotti contaminati nella fi liera di produzione degli alimenti. L’obiettivo del presente lavoro è stato quello di valutare il possibile uso degli emosieri testicolari ottenibili al momento della castrazione dei suinetti (processing fl uids, PF) per la ricerca di anticorpi anti-HEV. Cinquantacinque campioni individuali appaiati di siero e PF sono stati prelevati da altrettanti suinetti appartenenti a 20 diverse nidiate e analizzati con un kit commerciale ELISA per la ricerca di anticorpi anti-HEV. Una correlazione positiva (rho=0,677; p2,5) o alto (S/Co>6) livello anticorpale è miscelato con 5 o 6 campioni individuali di PF negativi. Il rilevamento, nei PF, di anticorpi anti-HEV di origine materna indica una passata esposizione al virus delle scrofe. I PF potrebbero rappresentare uno strumento rapido, non-invasivo ed economico per identifi care le aziende infette da HEV. Hepatitis E virus (HEV) is an RNA virus causing an acute generally self-limited disease in humans. Pigs and wild boars are the main reservoirs. Identifying HEV positive pig farms is important to implement surveillance programmes for this emerging zoonotic agent and to limit the risk of introducing contaminated products into the food chain. The aim of this study was to evaluate the use of serosanguineous fl uids obtained as part of castration practice (processing fl uids, PF) to detect anti-HEV antibodies in newborn piglets. Fiftyfi ve paired serum and PF samples were collected from piglets of 20 different litters and tested with a commercial ELISA kit. A positive correlation (rho=0.677; p2.5) or strong (S/Co>6) antibody level is mixed with 5 or 6 individual negative PF samples. The detection of anti-HEV maternal-derived antibodies in PF confi rms a past exposure of sows to the virus. PF may represent a rapid, non-invasive and economical tool to identify HEV positive farms.

Hepatitis E virus (HEV), is the causative agent of hepatitis E in humans. The disease is an acute hepatitis, generally self-limited which can become chronic in immunosuppressed patients. Among the 8 genotypes of HEV belonging to Orthohepevirus A, genotypes HEV-1 and HEV-2 infect only humans; HEV-3 and HEV-4 are zoonotic for which domestic and feral pigs are the main reservoirs. In most cases, the origin of autochthonous Hepatitis E cases in humans is unknown, but foodborne transmission has been clearly associated to sporadic cases and small clusters of infection linked to the consumption of contaminated pig liver sausages, raw deer meat, or undercooked wild boar meat. In Europe as well as in Italy, HEV-3 strains are widespread in pig farms and the within-herd prevalence varies from 10 to 100% depending on the study. The infection among animals is transmitted through the fecal oral route but it is not clear how long the virus can persist in the farm and the role of environmental as source of infection. The farm investigated was a farrow-to-finish herd consisting of 1000 breeders (site 1, breeding and nursery production stage) with growing pigs located in another premise (site 2, finisher production stage) in Northern Italy, sampled three times across 15 months. A total of 281 pool fecal samples were collected in 2017 (n=99), 2018 (n=142) and 2019 (n=40). Total viral RNA was extracted from fecal suspensions, and the HEV genome was detected by quantitative Real-Time RT-PCR (RT-qPCR).

Introduction Group A rotaviruses (RVA) are the leading cause of acute gastroenteritis (AGE) in young children, causing ca 250,000 deaths worldwide, mainly in low-income countries. Two proteins, VP7 (glycoprotein, G genotype) and VP4 (protease-sensitive protein, P genotype), are the basis for the binary RVA nomenclature. Although 36 G types and 51 P types are presently known, most RVA infections in humans worldwide are related to five G/P combinations: G1P[8], G2P[4], G3P[8], G4P[8], G9P[8]. Aim This study aimed to characterise the RVA strains circulating in Italy in the pre-vaccination era, to define the trends of circulation of genotypes in the Italian paediatric population. Methods Between September 2014 and August 2017, after routine screening in hospital by commercial antigen detection kit, 2,202 rotavirus-positive samples were collected in Italy from children hospitalised with AGE; the viruses were genotyped following standard European protocols. Results This 3-year study revealed an overall predominance of the G12P[8] genotype (544 of 2,202 cases; 24.70%), followed by G9P[8] (535/2,202; 24.30%), G1P[8] (459/2,202; 20.84%) and G4P[8] (371/2,202; 16.85%). G2P[4] and G3P[8] genotypes were detected at low rates (3.32% and 3.09%, respectively). Mixed infections accounted for 6.49% of cases (143/2,202), uncommon RVA strains for 0.41% of cases (9/2,202). Conclusions The emergence of G12P[8] rotavirus in Italy, as in other countries, marks this genotype as the sixth most common human genotype. Continuous surveillance of RVA strains and monitoring of circulating genotypes are important for a better understanding of rotavirus evolution and genotype distribution, particularly regarding strains that may emerge from reassortment events.

Over the last 10 years in Europe an increasing number of autochthonous cases linked to the zoonotic hepatitis E virus (HEV-3) have been reported. Pigs and wild boars are the main reservoirs. Human cases have been linked to consumption of raw or undercooked pork liver sausages and wild boar meat. In this study, we investigated 116 wild boar paired liver and muscle samples collected in the hunting season 2017-2018 (Viterbo). HEV RNA was detected in 14 (12.1%) livers of animals, 6 of which were also positive in muscles. Sequence analysis showed the presence of the same strains in muscle and liver. HEV strains sequenced clustered within subtypes HEV-3c, HEV-3f and other clusters not assignable to any subtypes described so far. Results obtained may suggest the risk of consuming raw or undercooked wild boar liver and meat. Furthermore, the presence of several HEV strains in the wild boar population investigated highlights the importance of the wild animals as reservoir of the zoonotic HEV-3.

Hepatitis E virus (HEV) is an RNA virus causing an acute generally self-limited disease in humans. Over the last 10 years in Europe an increasing number of autochthonous cases linked to foodborne transmission of HEV genotype 3 have been reported. Pigs and wild boars are considered the main reservoirs. Human cases have been frequently linked to the consumption of raw or undercooked pork products and wild boar meat. In this study, we sampled 92 and 116 wild boar (Sus scrofa) livers during the hunting seasons 2016-2017 and 2017-2018, respectively. HEV RNA was detected in 62 livers with prevalence ranging from 0.0% to 65.7% depending on the hunting area. Phylogenetic analysis showed that strains clustered within the different subtypes HEV-3c, HEV-3f and other clusters not assignable to any subtypes described so far. The latter could be considered novel subtype. The main result of sequencing was the wide hetherogeneneity of circulating strains, reporting the result to the confined area investigated (≈720 km2). Animals possibly belonging to the same family group hunted by the same team were infected with a unique strain (100% nucleotide identity). Since wild animals are a proven source of HEV transmission to humans, results obtained underlined the risk of consuming raw or undercooked wild boar meat and thus this subject deserves further investigations. Furthermore, the occurrence of such variable HEV strains in the wild boar population investigated highlights the importance of the wild animals as reservoir of the zoonotic HEV genotype.

During the last decade, an increasing number of autochthonous cases linked to the zoonotic genotype HEV-3, have been reported in Europe. The main transmission route is foodborne linked to the consumption of raw or undercooked pork products and wild boar meat. Pigs and wild boars are the main HEV-3 reservoirs. In this study, 92 wild boar (Sus scrofa) livers were sampled during the hunting season 2016-2017 in five municipalities in Central Italy. HEV RNA was detected in 52.2% of livers with prevalence ranging from 0.0% to 65.7% in 4 of 5 areas investigated. By sequence analysis, strains were classified into HEV-3c and HEV-3f subtypes. Several strains were circulating in the same area and strains infecting animals possibly belonging to the same family were identical. Since, wild animals are a proven source of HEV transmission to humans and pigs, the high prevalence observed poses a question on the risk of consuming wild boar meat.

International audience; We sought datasets with granular age distributions of rotavirus-positive disease presentations among children

In October 2017, two outbreaks of gastroenteritis (GE) occurred among patrons of a cafeteria in Italy in one week. Virological and bacteria investigations on stool samples, environment and food were conducted to identify the infectious agents and the possible source of infection. Forty-five cases occurred in the two outbreaks, including 13 laboratory-confirmed cases of norovirus GI. Nine staff members were interviewed, six were confirmed positive for NoV GI and 3 experienced GE symptoms. Bacteria faecal indicators and other bacteria pathogens were not detected in either environmental swab samples or food. A low level of NoV GII was detected in two environmental swab samples. The same GI.6 strain was identified in cases related to both outbreaks, suggesting a common source of infection. Since the two outbreaks occurred in one week, the NoV contamination could have persisted in the cafeteria. Furthermore, virological investigation revealed confirmed cases among food handlers who had worked at the cafeteria between and during the two outbreaks. Several studies highlighted the importance of excluding symptomatic food handlers to prevent contamination of foods and environment.

Group A rotaviruses (RVA) are one of the major cause of acute gastroenteritis (AGE) in young children, being responsible for up to 250.000 deaths worldwide, mostly in developing countries. The two outer capsid proteins VP7 (glycoprotein, G-genotype) and VP4 (protease-sensitive protein, P-genotype) are the basis for the binary RVA nomenclature. Although at least 36 G-types and 51 P-types of rotavirus are presently known, most RVA infections in humans, worldwide as well as in Italy, are related to six major G/P combinations: G1P[8], G2P[4], G3P[8], G4P[8], G9P[8] and G12P[8]. In November 2016, in the framework of the Italian 2016/17 rotavirus surveillance season, a total of 22 rotavirus-positive samples from hospitalized children presenting AGE symptoms were collected in a small area of Central Italy (Ancona, Marche). After genotyping, 3 samples presented the G9P[4] genotype. In order to better understand the origin of these uncommon RVA strains causing dehydrating diarrhea in three children, the strains RVA/Human-wt/ITA/AN18/2016/G9P[4], RVA/Human-wt/ITA/AN19/2016/G9P[4] and RVA/Human-wt/ITA/AN22/2016/G9P[4] were subjected to nucleotide sequencing of all the 11 gene segments to define their genomic constellation. Nucleotide sequencing revealed that the genomic constellation of the three strains was G9-P[4]-I2-R2-C2-M2-A2-N2-T2-E2-H2, highlighting human origin for all the gene segments investigated. The molecular characterization of RVAs and the continue monitoring of their circulation is needed to better define the epidemiology of these pathogen and to detect the emergence of viral variants presenting a high spreading potential in humans in the post-vaccination era.

Nell’uomo il virus dell’epatite E (HEV) causa un’epatite acuta in genere asintomatica, che può diventare cronica in pazienti immunosoppressi. Due dei quattro genotipi di HEV, i genotipi 3 e 4, specie Orthohepevirus A, sono considerati zoonotici, infettano l’uomo e diverse specie animali tra cui il suino, che viene considerato il serbatoio principale. I ceppi di HEV che infettano il ratto, appartengono ad un’altra specie, gli Orthohepevirus C (genotipo HEV-­‐C1), ma sono stati identificati anche ceppi virali vicini ai ceppi zoonotici di genotipo 3. I rotavirus di gruppo A (RVA) sono responsabili della maggior parte delle gastroenteriti acute stagionali in pazienti di età compresa tra 0 e 5 anni. Oltre gli umani, RVA infetta un’ampia varietà di mammiferi, inclusi i ratti. Il genoma di RVA è composto di 11 segmenti di dsRNA. Al fine di ottenere un’informazione dettagliata dei virus circolanti, dal 2008 è stato adottato un sistema di genotipizzazione che prevede l’assegnazione del genotipo a tutti i segmenti genomici di RVA. La presenza di HEV in specie diverse di ratti è stata descritta in tutto il mondo con prevalenze comprese tra 0,3 e 18%, per quanto riguarda la circolazione di RVA, invece, sono ancora pochi gli studi disponibili. Nel presente studio contenuti intestinali e campioni di fegato, provenienti da 40 ratti (Rattus rattus) catturati in prossimità di allevamenti suini in Nord Italia, sono stati analizzati per la ricerca dell’RNA virale di HEV e RVA. Il contenuto intestinale di un ratto (1/40, 2.5%) è risultato positivo per la presenza di RVA e uno per HEV, mentre tutti i campioni di fegato sono risultati negativi. Il sequenziamento e l’analisi filogenetica hanno confermato la presenza del genotipo HEV-­‐C1 di ratto, correlato ad altri ceppi di ratto identificati in America ma lontano dai ceppi di genotipo 3. Lo stesso tipo di indagine effettuata per RVA ha permesso di stabilire il genotipo G3-­‐P[3]-­‐I1-­‐R11-­‐C11-­‐M10-­‐ A22-­‐N18-­‐T14-­‐E18-­‐H13, il quale rappresenta una combinazione genomica caratterizzata dalla presenza di un gene di derivazione umana associato ad un genoma tipicamente animale. In conclusione, la presenza di HEV e RVA in ratti circolanti in prossimità degli allevamenti costituisce un rischio per l’uomo, questi infatti possono servire da o ospiti o serbatoi di virus potenzialmente patogeni.

Zoonotic viruses are thought to be the highest risk of epidemic diseases for next years. Among others, these also include rotavirus, agent of gastroenteritis in man and animals and hepatitis E virus (HEV), which causes acute hepatitis in humans and infect domestic and wild animals. Group A rotaviruses (RVA) infection is preferentially species-specific, heterologous rotavirus infections may also occur, including animal-tohuman virus transmission. Recently, a human RVA of rabbit origin was described in a child with gastroenteritis, and a bovine-like RVA was isolated from a laboratory rabbit . Hepatitis E is an emerging disease now recognized to be a zoonosis. Swine, wild boar, deer and rabbit are considered reservoirs of the zoonotic HEV genotypes 3 and 4. Recently HEV was detected in both farmed and wild rabbits in several areas of China, USA and France. The prevalence of serum antibodies against HEV ranges between 7% and 57% in young animals. Several evidences support a risk of zoonotic transmission of HEV from rabbits to humans, including experimental infection of non-human primates and swine with rabbit HEV and detection of HEV closely related to rabbit HEV in a man. HEV and RVA infections were studied in 21 farmed and 121 pet rabbits in Italy. Virus infection was investigated by serological analysis in sera and by detection of RNA in fecal samples. During 2013, a total of 121 rabbits attending veterinary examination were enrolled in this study. Sera and feces were collected from the same animals (except 7 fecal samples). Seventy-seven pets were asymptomatic, most others were affected by different pathologies. Between October 2013 and February 2014, 21 samples were collected from stud-mare rabbits affected by reproductive failure. Western blotting (WB). The capsid protein of an Italian gt3 swine HEV strain expressed in Sf9 cells from a recombinant baculovirus (BacΔ111ORF2HEV), the purified SA11 RV and crude extract of mock Sf9 (negative control) were subjected to WB. Sera were diluted 1:100. Immunocytochemistry. Sf9 cells infected with BacΔ111ORF2HEV and mock cells were fixed and incubated with rabbit sera (1:500). Replication foci were visualized by reaction with amino-ethylcarbazole. Reverse-transcription–polymerase chain reaction (RT–PCR) and DNA sequencing. RNA was extracted using Qiagen RNA-Easy-Mini kit. For both viruses, RT-PCRs were performed using the OneStep RT-PCR kit (Qiagen). For Rotavirus the NSP5 segment was amplified. For HEV an RT-PCR and a nested-PCR were conducted, amplifying a 300 bp ORF2 fragment. None of the 135 fecal samples, (114 pets and 21 farmed rabbits), showed presence of HEV genome. IgG against HEV were detected in 4 of 121 pet rabbits tested. Two seropositive animals were asymptomatic, one was affected by rhinitis. Rabbits showed no clinical signs of RVA infection. Twelve out of 52 sera recognized specific RVA proteins. Twelve samples resulted positive in RT-PCR, one was confirmed through sequence analyses. One animal was positive for both RVA and HEV IgG. None of the feces (farmed or pet rabbits) were positive for HEV genome. The seroprevalence in pet rabbits was 3.3% in this study, markedly lower than observed in farmed animals (36% in China and 55% in USA). The lower seroprevalence in our study might be explained with the different animal population; pets living in households have no contact with other animals, which reduces the possible HEV transmission through the fecal-oral route. The study of naturally occurring heterologous rotaviruses may help understand how rotavirus cross the host-species barrier and investigate the molecular determinants that control rotavirus host-species specificity and pathogenicity. Contact between pet rabbits and their owners, e.g. changing the litter, might favor animal-to-human transmission; similar risks occur in persons with occupational exposure to farmed rabbits. Possible foodborne transmission of rabbit HEV should also be evaluated.

International audience; The number of facilities that offer tumor treatment with particle therapy has been increasing substantially over the past decades. Ions are known for their enhanced effectiveness in killing tumor cells when compared with protontherapy. Such a characteristic is quantified through the RBE (relative biological effectiveness), which is a complex function of multiple parameters like cell line, cell cycle stage, radiation quality and irradiation conditions. Therefore, determining the value of RBE for every scenario is a challenging task that requires modeling to comply with the demands of a clinical environment.We developed a new model: nanoxTM (NAdosimetry and Oxidative stress). The nanoxTM model takes as input dosimetry quantities at multi-scale, starting from nanoscale, but also the production of radicals induced by water radiolysis. The cell survival predicted by NanoxTM for V79 cell line was compared to experimental results for photons, protons and carbon ions, and even others like argon ions. A good agreement was found in all cases. In particular, the model is able to describe the effectiveness of ions, including the overkill effect at higher LET values. Moreover, NanoxTM can reproduce the typical shoulder in cell survival curves. This was possible due to the introduction of the “non-local” events, through the chemical dose, which varies with LET. It is worthwhile to note that such results were obtained through the adjustment of a reduced number of free parameters.

International audience

Bivalve mollusks as filter-feeders concentrate in their digestive tissue microorganisms likely present in the harvesting water, thus becoming risky food especially if consumed raw or poorly cooked. To eliminate bacteria and viruses eventually accumulated, they must undergo a depuration process which efficacy on viruses is on debate. To better clarify the worth of the depuration process on virus elimination from mussels, in this study we investigated rotavirus kinetics of accumulation and depuration in Mytilus galloprovincialis experimentally contaminated. Depuration process was monitored for 9 days and virus residual presence and infectivity were evaluated by real time quantitative polymerase chain reaction, cell culture and electron microscopy at days 1, 2, 3, 5, 7, 9 of depuration. Variables like presence of ozone and of microalgae feeding were also analyzed as possible depuration enhancers. Results showed a two-phase virus removal kinetic with a high decrease in the first 24 h of depuration and 5 days necessary to completely remove rotavirus.

Enteric viruses represent a public health risk since they are involved in foodborne and waterborne outbreaks. The aim of this study was to investigate the circulation of select human enteric viruses — hepatitis A virus (HAV), noroviruses (NoVGI and NoVGII), adenovirus (AdV), rotavirus (RV), astrovirus (AsV), sapovirus (SaV) and aichivirus (AiV) – in wastewater and in environmental samples (groundwater, surface water and sediments) of Campania Region, southern Italy, with the goal of evaluating their circulation in the environment and to characterize the circulating strains by phylogenetic analysis. A total of 199 water and sediment samples were collected from rivers, dams, wells and wastewater treatment plants (WWTPs) in all provinces of the Campania Region and analysed by real-time qPCR for the detection of viruses, after a concentration step with organic flocculation. This study provides information on the qualitative and quantitative spread of human enteric viruses in the environment and their potential health risk. In our samples all the viral families were detected, with different prevalence. Among the investigated viruses, RV was the most prevalent and concentrated virus in surface water and sediments, with AsV being prevalent and concentrated in wastewater. The prevalence of viral pathogens was not influenced by season for almost all enteric viruses, except for NoVGII in WWTPs. Within the WWTPs, a significant reduction in the average concentration levels was observed for all viral targets from the influent to the effluent except for the HAV and AiV. The results of our study confirmed the importance of environmental surveillance that can represent a key tool for pollution control and human risk exposure.

Background Group A rotaviruses (RVA) are the leading cause of acute gastroenteritis (AGE) in young (aged

Gastrointestinal disease is frequent in pigs, and among the different etiological agents involved, viruses are considered the leading cause of infection in this animal species. Furthermore, about half of the newly identified swine pathogens are viruses, many of which may be transmitted to humans by direct contact or by indirect transmission pathways. In this study, the prevalence of astrovirus (AstV), group A rotavirus (RVA), norovirus (NoV) and hepatitis E virus (HEV) infections in pigs was investigated. During 2012-2014, 242 fecal samples were collected from pigs at different production stages (5 to 220 days old) on eight swine farms located in northern, central and southern Italy. Seven out of eight farms analyzed were positive for AstV, which was detected in 163 out of 242 (67.4%) samples and was the most prevalent virus; 61 of the 163 AstV-positive animals (37.4%) had diarrhea. HEV was detected on six farms and in 45 (18.6%) of the 242 samples analyzed. Twenty-three HEV-infected pigs had diarrhea (51.1%). A lower prevalence was observed for RVA, which was found in 10 of the 242 samples (4.1%) from three positive farms, and diarrhea was present only in six infected pigs (60.0%). No swine samples were found to be positive for NoV. Genetic diversity and phylogenetic relationships of some strains representative of the different viruses detected were investigated, confirming a wide heterogeneity of viral strains circulating among pigs.

Rotavirus (RV) infection causes watery diarrhea via multiple mechanisms, primarily chloride secretion in intestinal epithelial cell. The chloride secretion largely depends on non-structural protein 4 (NSP4) enterotoxic activity in human enterocytes.through mechanisms that have not been defined. Redox imbalance is a common event in cells infected by viruses, but the role of oxidative stress in RV infection is unknown. RV SA11 induced chloride secretion in association with an increase in reactive oxygen species (ROS) in Caco-2 cells. The ratio between reduced (GSH) and oxidized (GSSG) glutathione was decreased by RV. The same effects were observed when purified NSP4 was added to Caco-2 cells. N-acetylcysteine (NAC), a potent antioxidant, strongly inhibited the increase in ROS and GSH imbalance. These results suggest a link between oxidative stress and RV-induced diarrhea. Because Saccharomyces boulardii (Sb) has been effectively used to treat RV diarrhea, we tested its effects on RV-infected cells. Sb supernatant prevented RV-induced oxidative stress and strongly inhibited chloride secretion in Caco-2 cells. These results were confirmed in an organ culture model using human intestinal biopsies, demonstrating that chloride secretion induced by RV-NSP4 is oxidative stress-dependent and is inhibited by Sb, which produces soluble metabolites that prevent oxidative stress. The results of this study provide novel insights into RV-induced diarrhea and the efficacy of probiotics.

Per la Malattia di Newcastle (ND) e la Rinotracheite del tacchino (TRT), vaccini vivi attenuati vengono comunemente somministrati nei primi giorni di vita ad almeno una settimana di distanza l’uno dall’altro per il timore che si verifichino interferenze tra virus vaccinali. La possibilità di associare in incubatoio tali interventi avrebbe numerosi vantaggi sia di ordine pratico, che economico e sanitario. In quest’ottica diventa fondamentale assicurarsi che non ci siano interferenze negative tali da compromettere l'efficacia delle vaccinazioni o addirittura causare effetti patologici indesiderati. L’obiettivo del presente lavoro è stato valutare, in condizioni sperimentali, l'interferenza fra due diversi ceppi vaccinali di NDV e uno di Metapneumovirus aviare (AMPV), agente della TRT, somministrati nel tacchino in associazione ad un giorno di vita, secondo un possibile schema vaccinale praticabile in incubatoio. La prova è stata eseguita in condizioni di isolamento biologico in isolatori per pollame. Ottanta tacchinotti commerciali sono stati suddivisi in gruppi e vaccinati al primo giorno di vita per AMPV col ceppo VCO3, e per NDV con i ceppi B1 o VG/GA, somministrati singolarmente o in associazione (VCO3 + B1 o VCO3+VG/GA). Nei 30 giorni successivi, ad intervalli stabiliti, è stata valutata l’eliminazione respiratoria dei virus vaccinali mediante real time RT-PCR, e la risposta anticorpale umorale per NDV ed AMPV, mediante inibizione dell’emoagglutinazione ed ELISA, rispettivamente. Infine, parte dei soggetti vaccinati per AMPV, singolarmente o in associazione, sono stati infettati sperimentalmente con AMPV allo scopo di valutare la protezione immunitaria mediante misurazione della sintomatologia clinica. I risultati ottenuti permettono di concludere che la somministrazione al tacchino di un giorno di vita di vaccino vivo attenuato AMPV ceppo VC03, in associazione con i vaccini vivi NDV sia ceppo B1 sia ceppo VG/GA, conferisce protezione al challenge con AMPV sovrapponibile a quella evocata dal vaccino somministrato singolarmente, e non determina interferenze negative sulla replicazione dei virus vaccinali a livello respiratorio e sulla risposta immunitaria umorale. Resta da verificare se la protezione nei riguardi di NDV a seguito di vaccinazione combinata sia sovrapponibile a quella ottenibile con la vaccinazione singola.

Nelle prime due settimane di gennaio 2011, circa 3.000 carcasse di tortore dal collare (Streptopelia decaocto) a vita libera sono state rinvenute nei pressi di un sito industriale, in provincia di Ravenna, dedito alla lavorazione di partite di semi o loro residui. L’esame autoptico di 322 soggetti evidenziava prevalentemente un grave danno renale, epatomegalia e ipoplasia splenica. Gli esami istologici a livello del rene mostravano nefrosi associata a necrosi tubulari e una nefrite interstiziale linfoplasmacellulare. Inoltre, si osservavano deplezione linfocitaria nella milza unitamente ad emosiderosi splenica ed epatica. La stima dell’età su 46 soggetti identificava il 78% di adulti e il 22% di giovani (entro il primo anno di vita). Le analisi chimiche sulle ingesta e su pool di fegati non evidenziavano tossicità da pesticidi clorurati e fosforati, carbammati, triazine, piretroidi, stricnina, neonicotinoidi, Pb, Cd, Cr, Hg, Zn, Cu, micotossine, perossidi. Indagini molecolari (RT-PCR Real-Time) escludevano la presenza di virus influenzali, mentre veniva evidenziato RNA di Paramyxovirus aviario tipo 1 (APMV-1) tramite RTPCR in 176 su 193 animali analizzati. L’isolamento di 27 ceppi di APMV-1 è stato ottenuto inoculando pool di organi su uova embrionate di pollo SPF. Tali ceppi risultavano patogeni in base alla sequenza aminoacidica del sito di clivaggio della proteina di fusione. Il sequenziamento parziale del gene F ha permesso di identificare 2 differenti genotipi cocircolanti: il lineaggio 4b (APMV-1 ceppo piccione) e un distinto cluster genetico del lineaggio 4.