Biological Containment for African Swine Fever (ASF) Laboratories and Animal Facilities: The Italian Challenge in Bridging the Present Regulatory Gap and Enhancing Biosafety and Biosecurity Measures.

The African Swine Fever Virus (ASFV) is a DNA virus of the Asfarviridae family, Asfivirus genus. It is responsible for massive losses in pig populations and drastic direct and indirect economic impacts. The ever-growing handling of ASFV pathological material in laboratories, necessary for either diagnostic or research activities, requires particular attention to avoid accidental virus release from laboratories and its detrimental economic and environmental effects. Recently, the Commission Delegated Regulation (EU) 2020/689 of 17 December 2019 repealed the Commission Decision of 26 May 2003 reporting an ASF diagnostic manual (2003/422/EC) with the minimum and supplementary requirements for ASF laboratories. This decision generated a regulatory gap that has not been addressed yet. This paper aims to describe the Italian National Reference Laboratory (NRL) efforts to develop an effective and reliable biological containment tool for ASF laboratories and animal facilities. The tool consists of comprehensive and harmonized structural and procedural requirements for ASF laboratories and animal facilities that have been developed based on both current and repealed legislation, further entailing a risk assessment and internal audit as indispensable tools to design, adjust, and improve biological containment measures.

False memory in between-language conditions: a brief review on the effect of encoding and retrieving in different languages.

False memories have been extensively investigated over the past few decades using the Deese/Roediger-McDermott (DRM) paradigm. In this paradigm, participants study lists of words associatively related to a non-presented critical lure. During a memory test, these critical lures are falsely recalled or recognized. Most studies have focused on false memories that arise when both encoding and retrieval are conducted in the same language (i.e., within-language conditions), which is typically the participant's native or first language (L1). However, much less is known about false memories when critical lures appear in the memory test in a different language than the studied lists (i.e., between-language conditions), being one of them the participant's second language (L2). The main objective of this exhaustive review was to provide an overview of the current state of research on false recognition using the DRM paradigm in between-language conditions, where languages are switched between encoding and retrieval (i.e., L1L2 versus L2L1). The results revealed a language dominance effect in between-language false memories. In other words, false recognition rates were dependent on the study language, with a trend toward higher false recognition when words were enconded in the L1 (L1L2) compared to when words were encoded in the L2 (L2L1). This review enhances our understanding of how studying words in a first or second language affects false memory in the DRM paradigm, emphasizing the significance of investigating false memory in second language speakers and the necessity for further research in the field.

False memory in a second language: The importance of controlling the knowledge of word meaning.

In the globalized world we live in, it is increasingly common for people to speak more than one language. Although research in psychology has been widely interested in the study of false memories with the Deese/Roediger-McDermott (DRM) paradigm, to date, there is a scarcity of studies comparing false memories in the first and the second language (L1 and L2, respectively). It is noteworthy that one of the most studied variables in the DRM paradigm, the backward associative strength (BAS), has hardly been studied in the L2. Moreover, the only study that recently examined this matter found differences in the knowledge of L2-word meaning between the high-BAS and low-BAS lists, which would hinder the interpretation of the BAS effect in L2 false memories. Taking all this into account, the current work examined false memories in the L1 (Spanish) and the L2 (English) as a function of BAS overcoming the limitations of the previous study. We selected DRM lists using both Spanish and English free association norms and lists were constructed to vary in BAS values while controlling the knowledge of word meaning. Results showed that false recognition was greater in the L1 or dominant language than in the L2 or non-dominant language. Furthermore, BAS modulated the false recognition in both the L1 and the L2. That is, false recognition was higher in high-BAS than low-BAS lists in both languages. Sensitivity index from the signal-detection theory helped us gain further insight into these results. The main findings are discussed in the light of theoretical models from both the false memory and the second language processing literature. Finally, practical implications and future research are provided.

"Choose it, and remember it": The impact of choice on destination memory.

Destination memory can be defined as the capacity to remember to whom we transmit information. It is measured through the accuracy of retrieving the association between the information we transmit and the person to whom we transmit it. A destination memory procedure aims to emulate human interaction by sharing facts with celebrities (i.e., familiar faces) since we often communicate with people we know. However, the role of the choice about who we intend to transmit the information to has not been evaluated before. This paper investigated whether deciding with whom to share a piece of information benefits destination memory. We designed two experiments with different levels of cognitive load, increasing it from Experiment 1 to Experiment 2. The experiments included two conditions: the choice condition, in which participants chose from two options to whom they desired to share a fact, and the no-choice condition, in which participants simply shared facts with celebrities without the possibility of a choice. Experiment 1 suggested that a choice component did not affect destination memory. However, when in Experiment 2 we raised the cognitive load by increasing the number of stimuli, we found that selecting the recipient during the more challenging task provided an advantage in destination memory. This result is congruent with the explanation that the shift of the participants' attentional resources to the recipient, caused by the choice component, leads to a destination memory improvement. In sum, it seems that a choice component can improve destination memory only under demanding attentional conditions.

Disentangling the Effects of Backward/Forward Associative Strength and Theme Identifiability in False Memory

BACKGROUND: False memory has been extensively studied using the Deese/Roediger-McDermott paradigm. Despite the robustness of the effect, there is wide variability in the results, which is not fully understood. METHOD: Three experiments independently examined the role of backward associative strength (BAS), forward associative strength (FAS), and theme identifiability (ID) on false memories. In Experiment 1, lists varied in BAS while controlling FAS and ID. In Experiment 2, FAS was manipulated while BAS and ID were controlled. Finally, in Experiment 3, lists varied in ID while controlling BAS and FAS. Data was analyzed using both frequentist and Bayesian analyses. RESULTS: We found false memories in all three experiments. Specifically, false recognition was higher in high-BAS than in low-BAS lists in Experiment 1. In Experiment 2, false recognition was higher in high-FAS than in low-FAS lists. In Experiment 3, false recognition was lower in high-ID than in low-ID lists. CONCLUSIONS: These findings suggest that both BAS and FAS─variables that promote error-inflating processes─and ID─which promotes error-editing processes─contribute independently to the production of false memories. Splitting apart the role of these variables helps to understand the variability of false memories and to extrapolate DRM tasks to explore other cognitive domains.

Disentangling the Effects of Backward/Forward Associative Strength and Theme Identifiability in False Memory / Aislando el Papel de la Fuerza Asociativa Directa e Inversa y la Identificabilidad del Tema Sobre los Recuerdos Falsos

Background: False memory has been extensively studied using the Deese/Roediger-McDermott paradigm. Despitethe robustness of the effect, there is wide variability in the results, which is not fully understood. Method: Threeexperiments independently examined the role of backward associative strength (BAS), forward associative strength(FAS), and theme identifiability (ID) on false memories. In Experiment 1, lists varied in BAS while controlling FAS andID. In Experiment 2, FAS was manipulated while BAS and ID were controlled. Finally, in Experiment 3, lists variedin ID while controlling BAS and FAS. Data was analyzed using both frequentist and Bayesian analyses. Results: Wefound false memories in all three experiments. Specifically, false recognition was higher in high-BAS than in low-BASlists in Experiment 1. In Experiment 2, false recognition was higher in high-FAS than in low-FAS lists. In Experiment3, false recognition was lower in high-ID than in low-ID lists. Conclusions: These findings suggest that both BAS andFAS—variables that promote error-inflating processes—and ID—which promotes error-editing processes—contributeindependently to the production of false memories. Splitting apart the role of these variables helps to understand thevariability of false memories and to extrapolate DRM tasks to explore other cognitive domains.(AU)

Antecedentes: las memorias falsas se han estudiado ampliamente utilizando el paradigma Deese/Roediger-McDermott. A pesar de la robustez del efecto, existe una amplia variabilidad de resultados que todavía no se comprendecompletamente. Método: tres experimentos examinaron independientemente el papel de la fuerza asociativa inversa(BAS), fuerza asociativa directa (FAS) e identificabilidad del tema (ID) en el reconocimiento falso (RF). Primero, semanipuló el BAS mientras se controló FAS e ID (Experimento 1). Segundo, se manipuló el FAS mientras se controlóBAS e ID (Experimento 2). Finalmente, se manipuló ID mientras se controló BAS y FAS (Experimento 3). Se utilizaronanálisis frecuentistas y bayesianos. Resultados: el RF fue mayor en las listas de alto que bajo BAS (Experimento 1), yalto que bajo FAS (Experimento 2). En cambio, el RF fue menor en las listas de alto ID que bajo ID (Experimento 3).Conclusiones: tanto BAS como FAS, variables que promueven procesos de inflación del error, pero también ID, quienpromueve procesos de edición del error, contribuyen de forma independiente a la producción de memorias falsas. Aislarel papel de estas variables ayuda a comprender la variabilidad de los falsos recuerdos y a extrapolar las tareas DRM paraexplorar otros dominios cognitivos.(AU)

Carnivore protoparvovirus 1 (CPV-2 and FPV) Circulating in Wild Carnivores and in Puppies Illegally Imported into North-Eastern Italy.

The illegal trade of animals poses several health issues to the global community, among which are the underestimated risk for spillover infection and the potential for an epizootic in both wildlife and domestic naïve populations. We herein describe the genetic and antigenic characterization of viruses of the specie Carnivore protoparvovirus 1 detected at high prevalence in puppies illegally introduced in North Eastern Italy and compared them with those circulating in wild carnivores from the same area. We found evidence of a wide diversity of canine parvoviruses (CPV-2) belonging to different antigenic types in illegally imported pups. In wildlife, we found a high circulation of feline parvovirus (FPV) in golden jackals and badgers, whereas CPV-2 was observed in one wolf only. Although supporting a possible spillover event, the low representation of wolf samples in the present study prevented us from inferring the origin, prevalence and viral diversity of the viruses circulating in this species. Therefore, we suggest performing more thorough investigations before excluding endemic CPV-2 circulation in this species.

Disinfectants against African Swine Fever: An Updated Review.

African Swine Fever (ASF), a hemorrhagic disease with a high mortality rate in suids, is transmitted via direct and indirect contact with infectious animals and contaminated fomites, respectively. ASF reached Europe in 2014, affecting 14 of the 27 EU countries including, recently, the Italian peninsula. The fast and unprecedented spread of ASF in the EU has highlighted gaps in knowledge regarding transmission mechanisms. Fomites, such as contaminated clothing and footwear, farming tools, equipment and vehicles have been widely reported in the spread of ASF. The absence of available vaccines renders biosecurity measures, cleaning and disinfection procedures an essential control tool, to a greater degree than the others, for the prevention of primary and secondary introductions of ASF in pig farms. In this review, available data on the virucidal activity of chemical compounds as disinfectants against the ASF virus (ASFV) are summarized together with laboratory methods adopted to assess the virucidal activity.

Molecular Investigation of Recent Canine Parvovirus-2 (CPV-2) in Italy Revealed Distinct Clustering.

Canine parvovirus Type 2 (CPV-2) is a worldwide distributed virus considered the major cause of viral gastroenteritis in dogs. Studies on Italian CPV-2 are restricted to viruses circulating until 2017. Only one study provided more updated information on CPV-2 but was limited to the Sicily region. No information regarding the circulation and genetic characteristics of CPV-2 in Northeast Italy has been made available since 2015. The present study investigated the genetic characteristics of CPV-2 circulating in the dog population of Northeast Italy between 2013 and 2019. The VP2 gene of 67 CPV-2 was sequenced, and phylogenetic analysis was performed to identify patterns of distribution. Phylogenetic and molecular analysis highlighted unique characteristics of Northeast Italian CPV-2 and interestingly depicted typical genetic clustering of the Italian CPV-2 strains, showing the existence of distinct CPV-2 genetic groups. Such analysis provided insights into the origin of some Italian CPV-2 genetic clusters, revealing potential introductions from East European countries and the spread of CPV-2 from South/Central to North Italy. This is the first report that describes the genetic characteristics of recent Italian CPV-2. Tracking the genetic characteristics of CPV-2 nationally and globally may have impact on understanding the evolution and distribution of CPV-2, in particular in light of the current humanitarian emergency involving Ukraine, with the massive and uncontrolled movement of people and pet animals.

Swine Norovirus: Past, Present, and Future.

Norovirus, an ssRNA + virus of the family Caliciviridae, is a leading disease burden in humans worldwide, causing an estimated 600 million cases of acute gastroenteritis every year. Since the discovery of norovirus in the faeces of swine in Japan in the 1990s, swine norovirus has been reported in several countries on several continents. The identification of the human-associated GII.4 genotype in swine has raised questions about this animal species as a reservoir of norovirus with zoonotic potential, even if species-specific P-types are usually detected in swine. This review summarises the available data regarding the geographic distribution of norovirus in swine, the years of detection, the genotype characterisation, and the prevalence in specific production groups. Furthermore, we discuss the major bottlenecks for the detection and characterisation of swine noroviruses.

Clinical and pathological features of Merkel cell carcinoma: A 4-year follow-up observational retrospective study in Spain.

BACKGROUND: Merkel cell carcinoma (MCC) is a malignant skin cancer with a 5-year survival rate of approximately 50%. Knowledge of MCC has increased in recent years mostly due to improved diagnosis techniques. In Spain there is lack of information regarding the incidence and tumour characteristics, and the treatment approaches are not standardised. The objective of this study was to provide information of the clinical and epidemiological characteristics of MCC patients in Spain. METHODS: Retrospective, observational study involving 192 patients from 25 Spanish hospitals. Evaluated variables included overall survival and incidence rate of Merkel cell polyomavirus, in patients diagnosed from 2012 to 2016. RESULTS: The Spanish incidence rate was estimated 0.32/100,000 inhabitants/year, with variations according to geographical regions, being slightly higher in areas with greater sunlight exposure. In total, 61.5% of tumours showed expansive growth (progressive growth of the tumour), 78.6% showed localisation in UV-exposed skin. 97.4% of patients were diagnosed by excisional biopsy. Surgery was the first line treatment in 96.6% of patients, radiotherapy in 24.6%, and chemotherapy in 6.3%. These treatments were not mutually exclusive. Median overall survival was 38.3 months (78.4% at 12 months and 60% at 24 months). MCPyV was present in 33.8% of patients. CONCLUSION: The incidence of MCC in Spain is one of the highest in Europe, with a slight predominance in men. The sample has shown that a biopsy is available for diagnosis in most cases. Moreover, the treatment is surgical when the tumour is localized and is associated with lymphadenectomy, and/or it is radiotherapy if widespread.

Feelings of Contrast at Test Reduce False Memory in the Deese/Roediger-McDermott Paradigm.

False memories in the Deese/Roediger-McDermott (DRM) paradigm are explained in terms of the interplay between error-inflating and error-editing (e.g., monitoring) mechanisms. In this study, we focused on disqualifying monitoring, a decision process that helps to reject false memories through the recollection of collateral information (i.e., recall-to-reject strategies). Participants engage in recall-to-reject strategies using one or two metacognitive processes: (1) applying the logic of mutual exclusivity or (2) experiencing feelings of contrast between studied items and unstudied lures. We aimed to provide, for the first time in the DRM literature, evidence favorable to the existence of a recall-to-reject strategy based on the experience of feelings of contrast. One hundred and forty participants studied six-word DRM lists (e.g., spy, hell, fist, fight, abduction, mortal), simultaneously associated with three critical lures (e.g., WAR, BAD, FEAR). Lists differed in their ease to identify their critical lures (extremely low-BAS lists vs. high-BAS lists). At recognition test, participants saw either one or the three critical lures of the lists. Participants in the three-critical-lure condition were expected to increase their monitoring, as they would experience stronger feelings of contrast than the participants in the one-critical-lure condition. Results supported our hypothesis, showing lower false recognition in the three-critical-lure condition than in the one-critical-lure condition. Critically, in the three-critical-lure condition, participants reduced even more false memory when they could also resort to another monitoring strategy (i.e., identify-to-reject). These findings suggest that, in the DRM context, disqualifying monitoring could be guided by experiencing feelings of contrast between different types of words.

Questioning the Role of Forward Associative Strength in False Memories: Evidence From Deese-Roediger-McDermott Lists With Three Critical Lures.

We report an experiment examining the factors that produce false recognition in the Deese-Roediger-McDermott (DRM) paradigm. We selectively manipulated the probability that critical lures produce study items in free association, known as forward associative strength (FAS), while controlling the probability that study items produce critical lures in free association, known as backward associative strength (BAS). Results showed that false recognition of critical lures failed to differ between strong and weak FAS conditions. Follow-up correlational analyses further supported this outcome, showing that FAS was not correlated with false recognition, despite substantial variability in both variables across our stimulus sets. However, these correlational analyses did produce a significant and strong relationship between BAS and false recognition. These results support views that propose false memory is produced by activation spreading from study items to critical lures during encoding, which leads critical lures to be confused with episodically-experienced events.

The Role of Language Proficiency in False Memory: A Mini Review.

Memory errors and, specifically, false memories in the Deese/Roediger-McDermott paradigm have been extensively studied in the past decades. Most studies have investigated false memory in monolinguals' native or first language (L1), but interest has also grown in examining false memories in participants' second language (L2) with different proficiency levels. The main purpose of this manuscript is to review the current state of knowledge on the role of language proficiency on false memories when participants encode and retrieve information in the same language. To do so, a systematic literature search was conducted, and the available studies were reviewed. These studies differed in, for example, age, language proficiency, or material characteristics, including both high and low associative strength lists, and they reported different results. In this review, we attempted to make sense of the apparently contradictory results by carefully identifying participants' language dominance and L2 proficiency. Specifically, the results indicated that, first, people are more prone to produce false memories in their dominant than in their non-dominant language. This result generalizes to lists with high and low associative strength, as well as to participants of different ages. Second, false memories do not differ between two languages when speakers are equally proficient in both languages. Finally, highly proficient L2 speakers produce more false memories in their L2 than speakers with lower L2 proficiency. The results of this review will be considered in the light of the theoretical frameworks of false memories and bilingual language processing.

The effect of language proficiency and associative strength on false memory.

In two experiments we examined the role of language proficiency and associative strength in the production of false memory. We constructed Deese-Roediger-McDermott lists using both Spanish and English free association norms. Lists were constructed to vary in backward associative strength (BAS). Experiment 1 participants were native Spanish speakers with some proficiency in English while Experiment 2 participants were native Spanish participants that had either high, intermediate, or low English proficiency. Results showed that, in both Experiment 1 and Experiment 2, false recognition was greater in participants' dominant language (L1 or Spanish) than in their non-dominant language (L2 or English), and false recognition in L2 increased with L2 proficiency when low-BAS lists were studied (Experiment 2). Further, false recognition was higher in high-BAS lists than in low-BAS lists in both L1 and L2. Finally, we collected a measure of participants' knowledge of our stimulus words in L2. These data showed that participants had far from perfect knowledge of all L2 stimuli. Analyses that factored out the effects of L2 word knowledge failed to alter the effects of L1 vs. L2, L2 proficiency and BAS on false recognition.

Genetic Variability among Swine Influenza Viruses in Italy: Data Analysis of the Period 2017-2020.

Swine play an important role in the ecology of influenza A viruses (IAVs), acting as mixing vessels. Swine (sw) IAVs of H1N1 (including H1N1pdm09), H3N2, and H1N2 subtypes are enzootic in pigs globally, with different geographic distributions. This study investigated the genetic diversity of swIAVs detected during passive surveillance of pig farms in Northern Italy between 2017 and 2020. A total of 672 samples, IAV-positive according to RT-PCR, were subtyped by multiplex RT-PCR. A selection of strains was fully sequenced. High genotypic diversity was detected among the H1N1 and H1N2 strains, while the H3N2 strains showed a stable genetic pattern. The hemagglutinin of the H1Nx swIAVs belonged to HA-1A, HA-1B, and HA-1C lineages. Increasing variability was found in HA-1C strains with the circulation of HA-1C.2, HA-1C.2.1 and HA-1C.2.2 sublineages. Amino acid deletions in the HA-1C receptor binding site were observed and antigenic drift was confirmed. HA-1B strains were mostly represented by the Δ146-147 Italian lineage HA-1B.1.2.2, in combination with the 1990s human-derived NA gene. One antigenic variant cluster in HA-1A strains was identified in 2020. SwIAV circulation in pigs must be monitored continuously since the IAVs' evolution could generate strains with zoonotic potential.

Interface between Bats and Pigs in Heavy Pig Production.

Bats are often claimed to be a major source for future viral epidemics, as they are associated with several viruses with zoonotic potential. Here we describe the presence and biodiversity of bats associated with intensive pig farms devoted to the production of heavy pigs in northern Italy. Since chiropters or signs of their presence were not found within animal shelters in our study area, we suggest that fecal viruses with high environmental resistance have the highest likelihood for spillover through indirect transmission. In turn, we investigated the circulation of mammalian orthoreoviruses (MRVs), coronaviruses (CoVs) and astroviruses (AstVs) in pigs and bats sharing the same environment. Results of our preliminary study did not show any bat virus in pigs suggesting that spillover from these animals is rare. However, several AstVs, CoVs and MRVs circulated undetected in pigs. Among those, one MRV was a reassortant strain carrying viral genes likely acquired from bats. On the other hand, we found a swine AstV and a MRV strain carrying swine genes in bat guano, indicating that viral exchange at the bat-pig interface might occur more frequently from pigs to bats rather than the other way around. Considering the indoor farming system as the most common system in the European Union (EU), preventive measures should focus on biosecurity rather than displacement of bats, which are protected throughout the EU and provide critical ecosystem services for rural settings.

Divergent minute virus of canines strains identified in illegally imported puppies in Italy.

Minute virus of canines (MVC) belongs to the family Parvoviridae, genus Bocaparvovirus, and has been mainly described during enteritis episodes in young dogs. This study reports the characterization of four divergent MVC strains detected between 2012 and 2018, three of which were from dogs illegally imported into Italy, most probably from Eastern Europe, that cluster together phylogenetically but share low genetic similarity with the fourth MVC from an autochthonous dog and other available MVC sequences. Our data indicate that the introduction of genetically distinct MVC strains occurred through the illegal movement of dogs from a geographic area where a distinct MVC lineage was most likely circulating. Enforced surveillance of MVC in the dog population of Eastern Europe and its neighboring countries may shed light on, and eventually trace back to, illegal animal movements.

Evidence of ongoing complement activation on adipose tissue from an 11-year-old girl with Barraquer-Simons syndrome.

Barraquer-Simons syndrome (BSS), a form of acquired partial lipodystrophy, is a rare condition characterized by gradual loss of adipose tissue from the upper body, keeping intact the white adipose tissue of the lower extremities. The etiology of BSS is not well understood, and clinical follow-up studies have not been assessed in these patients. Moreover, no histological studies have been conducted during the active phase of the disease, and complement system activation products have not been sought in the affected areas. The objective of this work was to analyze the clinical, immunological and histological events in an 11-year-old girl with BSS over a 5-year follow-up period. Clinical data were collected during six regular visits for a time period of 5 years. The circulating levels of C3, C3adesArg (a product released upon C3 activation), C4 and immunoglobulins (Ig) were quantified in serum while fat tissue from lipoatrophic areas was examined by immunohistochemical and immunofluorescence approaches. In her regular visits, no clinical or laboratory abnormalities had been observed in the patient, except for the progression of lipoatrophy linked to the C3 hypocomplementemia and the occurrence of C3 nephritic factor. Adipose tissue from the patient showed atrophied and dead adipocytes, an abnormal production of extracellular matrix, and a remarkable accumulation of infiltrating CD68 macrophages and adipocyte precursors (marked by c-Kit positiveness). Simultaneous detection of IgG, C3, C5a and C5b-9 proved the ongoing complement activity and complement-directed injury within the adipose tissue. Our results showed the first evidence that the complement system hyperactivation occurs within the adipose tissue and is linked with fat loss in patients with BSS.