A web-based geographic information system monitoring wildlife diseases in Abruzzo and Molise regions, Southern Italy.

BACKGROUND: Nowadays there is a worldwide consensus on the importance of conducting wildlife disease surveillance. Indeed, 60% of emerging infectious diseases are zoonotic in nature, and the majority of these (71.8%) originate in wildlife. Surveillance of wildlife diseases is crucial to prevent negative effects on human and animal health. Data digitization and sharing are among the main goals for the present and coming years. Geographic Information Systems (GIS) are increasingly used to analyze the geographical distribution of diseases and the relationships between pathogenic factors and their geographic environments. METHODS: Wild animal's samples collected in the Abruzzo and Molise regions and delivered to our laboratory are entered in our Laboratory Information System and processed to be displayed through the Web-GIS mash-up presented in this paper. We built it using both open source and proprietary solutions, to produce data driven interactive maps, charts and tables to help to understand the epidemiology of wild animal diseases, their spread and trend. RESULTS: Since 2013, 9.606 samples collected from wild animals have been analyzed in the laboratories of the IZS-Teramo and have been recorded in the system, facilitating the reporting to the judicial authorities and the identification of highly risky areas to set up control and repression measures. Moreover, thanks to the monitoring health protocol, a canine distemper epidemic in wolves has been detected and monitored in its temporal and spatial evolution, as well as cases of bovine tuberculosis in wild boars. CONCLUSIONS: While it is more evident that the starting point is to choose the right sampling method, it is for sure less obvious that the information system in which data is stored is equally important. In fact, it should give the possibility to consult it in an easy and instructive way. GIS allows immediately grasping the spatial relationships between the data itself and those between the data and the territory; it is an important tool to support veterinary services in managing epidemic and non-epidemic emergencies and performing epidemiological investigations, but also to examine control plans and identify new gaps and challenges.

Brucella abortus Strain RB51 Administered to Prepubescent Water Buffaloes, from Vaccination to Lactation: Kinetics of Antibody Response and Vaccine Safety.

Brucella RB51 is a live modified vaccine. Its use in water buffalo has been proposed using a vaccination protocol different to that used for cattle, but knowledge of the long-term effects of RB51 vaccination in this species remains incomplete. The aim of the study was to evaluate the safety and kinetics of antibody responses in water buffaloes vaccinated according to the protocol described for the bovine species in the WOAH Manual, modified with the use of a triple dose. Water buffaloes were vaccinated with the vaccine RB51. A booster vaccination was administered at 12 months of age. When turning 23-25 months old, female animals were induced to pregnancy. RB51-specific antibodies were detected and quantified using a CFT based on the RB51 antigen. Vaccinated animals showed a positive serological reaction following each vaccine injection, but titers and the duration of the antibody differed among animals. For 36 weeks after booster vaccination, the comparison of CFT values between vaccinated and control groups remained constantly significant. Afterwards, antibody titers decreased. No relevant changes in antibody response were recorded during pregnancy or lactation. In conclusion, results indicated that the vaccination schedule applied is safe and allows for vaccinated and unvaccinated controls to be discriminated between for up to 8 months after booster vaccination.

One health system supporting surveillance during COVID-19 epidemic in Abruzzo region, southern Italy.

The Istituti Zooprofilattici Sperimentali (IZSs) are public health institutes dealing with the aetiology and pathogenesis of infectious diseases of domestic and wild animals. During Coronavirus Disease 2019 epidemic, the Italian Ministry of Health appointed the IZSs to carry out diagnostic tests for the detection of SARS-CoV-2 in human samples. In particular, the IZS of Abruzzo and Molise (IZS-Teramo) was involved in the diagnosis of SARS-CoV-2 through testing nasopharyngeal swabs by Real Time RT-PCR. Activities and infrastructures were reorganised to the new priorities, in a "One Health" framework, based on interdisciplinary, laboratory promptness, accreditation of the test for the detection of the RNA of SARS-CoV-2 in human samples, and management of confidentiality of sensitive data. The laboratory information system - SILAB - was implemented with a One Health module for managing data of human origin, with tools for the automatic registration of information improving the quality of the data. Moreover, the "National Reference Centre for Whole Genome Sequencing of microbial pathogens - database and bioinformatics analysis" - GENPAT - formally established at the IZS-Teramo, developed bioinformatics workflows and IT dashboard with ad hoc surveillance tools to support the metagenomics-based SARS-CoV-2 surveillance, providing molecular sequencing analysis to quickly intercept the variants circulating in the area. This manuscript describes the One Health system developed by adapting and integrating both SILAB and GENPAT tools for supporting surveillance during COVID-19 epidemic in the Abruzzo region, southern Italy. The developed dashboard permits the health authorities to observe the SARS-CoV-2 spread in the region, and by combining spatio-temporal information with metagenomics provides early evidence for the identification of emerging space-time clusters of variants at the municipality level. The implementation of the One Health module was designed to be easily modelled and adapted for the management of other diseases and future hypothetical events of pandemic nature.

Estimation of the risk of fipronil ingestion through the consumption of contaminated table eggs for the Italian consumer.

Fipronil is an insecticide which is not approved for use in any food-producing animal species in the European Union (EU). However, the inappropriate use of fipronil in mites' disinfestation products utilized in poultry farms in the Netherlands and other EU countries in 2017, led to the detection of residues of this pesticide in eggs across Europe. In Italy, a national monitoring plan was established to verify the possible misuse of fipronil in Italian laying hens. Out of 577 sampled farms, 23 eggs resulted contaminated (4.0%; 95% CI: 2.7%-5.9%). A higher prevalence of contamination was observed in flocks kept on cage (8.7%; 95% CI: 6.0% -12.4%) than on ground (1.6%; 95% CI: 0.7% -3.7%); Chi-square = 16.1; P <0.001). The results allowed developing a stochastic model for estimating the risk of fipronil ingestion through the consumption of contaminated table eggs for the Italian consumer. The probability that an individual ingests a dose of fipronil greater than the acute reference dose (ARfD, equal to 0.009 mg/kg body weight) was assessed as very low, ranging from values very close to 0 in people with more than 10 years of age and 0.0007 in infants less than 3 years.

A COVID-19 Hotspot Area: Activities and Epidemiological Findings.

By late March 2020, Villa Caldari, a small village of the municipality of Ortona (Abruzzo region), was registering an incidence rate of COVID-19 cases ten times greater than the overall municipality and was declared a hotspot area. Twenty-two days later, epidemiological investigation and sampling were performed, to evaluate SARS-CoV-2 circulation and the presence of SARS-CoV-2 antibodies. Overall, 681 nasopharyngeal swabs and 667 blood samples were collected. Only one resident of the village resulted in being positive for RNA viral shedding, while 73 were positive for SARS-CoV-2 antibodies. The overall seroprevalence was 10.9%. The difference between the seroprevalence of infection in asymptomatic and symptomatic individuals was significant (χ2 = 14.50 p-value = 0.0001). Amongst the residents positive for antibodies, fatigue and/or muscle pain, fever and anosmia were the most experienced symptoms, whose most frequent onset was observed during the first two weeks of March. Familial and habit-related clusters were highlighted. Nevertheless, the investigations showed a low SARS-CoV-2 circulation in the village at the time of the sampling, demonstrating virus transmission could be limited when strict emergency measures are followed. Given the favorable results, the emergency measures were then lifted.

African Swine Fever: Lessons to Learn From Past Eradication Experiences. A Systematic Review.

Prevention, early detection, prompt reaction, and communication play a crucial role in African swine fever (ASF) control. Appropriate surveillance capable of early detection of the disease in both domestic and wild animals, and the implementation of consolidated contingency plans, are currently considered the best means of controlling this disease. The purpose of this study was to understand the lessons to be learned through the global disease eradication history. To establish which strategies were successful for prevention, control, and eradication of ASF, and which errors should not be repeated, we conducted a systematic review. A query was defined to search for surveillance and control strategies applied by countries worldwide for ASF eradication in the past. Inclusion and exclusion criteria were defined. Decisions on study eligibility and data extraction were performed by two independent reviewers and the differences were resolved by consensus or by a third reviewer. From 1,980 papers, 23 were selected and included in the qualitative analysis. Reports from Belgium, Brazil, Cuba, the Dominican Republic and Haiti, France, mainland Italy, Malta, Portugal, and Spain were included. Despite the economic resources allocated and the efforts made, eradication was possible in only eight countries, between the 50s and 90s in the twentieth century, in different epidemiological and cultural contexts, in some instances within <1 year, and in others in about 40 years. Classical surveillance strategies, such as active and passive surveillance, both at farm and slaughterhouse levels, targeted surveillance, together with conventional biosafety and sanitary measures, led to eradication even in countries in which the tick's epidemiological role was demonstrated. Historical surveillance data analysis indicated that eradication was possible even when technological tools either were not available or were used less than they are currently. This emphasizes that data on surveillance and on animal population are crucial for planning effective surveillance, and targeting proper control and intervention strategies. This paper demonstrates that some strategies applied in the past were effective; these could be implemented and improved to confront the current epidemiological wave. This offers encouragement for the efforts made particularly in Europe during the recent epidemics.

Campylobacter jejuni and Campylobacter coli: prevalence, contamination levels, genetic diversity and antibiotic resistance in Italy.

A research was carried out in Italy with the aim of assessing Campylobacter contamination in broilers from breeding to slaughter, of defining the genetic diversity of isolates and their antibiotic resistance. Sampling was carried out in a slaughterhouse, and in farms representative of the most common broiler production in Italy. At farm, the 78.8% (95% C.I.: 74.5%­82.5%) of cloacal samples tested positive for Campylobacter spp. C. jejuni showed higher prevalence in winter than in spring and summer (p < 0.00001, χ2 = 32.9), while C. coli showed an opposite trend (p < 0.00001, χ2= 41.1). At slaughterhouse, the 32.3% (95% C.I.: 30.2%­35.2%) and the 23.9% (95% C.I.: 21.7%­26.3%) of skin samples tested positive for C. jejuni for C. coli, respectively. C. coli showed higher prevalence than C. jejuni at washing (p < 0.05, χ2 = 11.11) and at chilling (p < 0.05, χ2 = 9.26). PFGE revealed high heterogeneity among isolates. Some clones were identified within the same farm in more than one season, suggesting environmental conditions able to support their persistence; other clones resulted to be spatially distant, suggestive of cross­contamination. Both Campylobacter species showed high resistance to nalidixic acid and ciprofloxacin, while resistance to erythromycin was more frequent in C. coli than C. jejuni (p < 0.05; χ2 test).

Multi-locus sequence typing and virulence profile in Bacillus cereus sensu lato strains isolated from dairy products.

Members of Bacillus cereus group are important food contaminants and they are of relevant interest in food safety and public heath due to their ability to cause two distinct forms of food poisoning, emetic and diarrhoeal syndrome. In the present study, 90 strains of B. cereus isolated from dairy products, have been typed using Multilocus Sequence Typing (MLST) analysis and investigated for the occurrence of 10 enterotoxigenic genes (hblA, hblC, hblD, nheA, nheB, nheC, cytK, entFM, entS and bceT) and one emetogenic gene (ces), to determine their genetic diversity. A total of 58 sequence types were identified and among these 17 were signalled as new profiles. Among the virulence genes, the majority of our strains carried the entS (92%), entFM (86%), nhe (82%) and cytK (72%) genes. All remaining genes were identified in at least one strain with different prevalence, stressing the genetic diversity, how even the different grade of pathogenicity of B. cereus isolated from dairy products.

Prevalence estimation of Italian ovine cystic echinococcosis in slaughterhouses: A retrospective Bayesian data analysis, 2010-2015.

Cystic echinococcosis (CE) is a complex zoonosis with domestic and sylvatic life-cycles, involving different intermediate and definitive host species. Many previous studies have highlighted the lack of a surveillance system for CE, its persistence in Italy, and endemicity in several Italian regions. Because of the absence of a uniform surveillance program for both humans and animals, disease occurrence is widely underestimated. This study aimed to estimate the prevalence of ovine CE in Italy. Survey data on the prevalence of Echinococcus granulosus complex infections in Italian sheep farms from 2010 to 2015 were obtained in collaboration with Regional Veterinary Epidemiology Observatories (OEVRs). Bayesian analysis was performed to estimate the true CE farm prevalence. The prior true CE prevalence was estimated using data from Sardinia. Second, Bayesian modelling of the observed prevalence in different regions and the true prevalence estimation from the first step were used to ultimately estimate the prevalence of ovine CE in Italy. We obtained survey data from 10 OEVRs, covering 14 Italian regions. We observed that the risk of CE infection decreased over the years, and it was strictly correlated with the density of susceptible species. Using Sardinia as prior distribution, where the disease farm prevalence was approximately 19% (95% CI, 18.82-20.02), we estimated that the highest endemic CE farm prevalence was in Basilicata with a value of 12% (95% BCI: 7.49-18.9%) and in Piemonte 7.64%(95% BCI: 4.12-13.04%). Our results provide spatially relevant data crucial for guiding CE control in Italy. Precise information on disease occurrence location would aid in the identification of priority areas for disease control implementation by the authorities. The current underestimation of CE occurrence should urge the Italian and European governments to become aware of the public health importance of CE and implement targeted interventions for high-risk areas.

Estimating the prevalence of Brucellosis in cattle in Zimbabwe from samples submitted to the Central Veterinary Laboratory between 2010 and 2014.

Brucellosis is a disease caused by bacteria belonging to the genus Brucella, which is endemic in sub-Saharan African countries, including Zimbabwe. Zimbabwe has a widespread dairy industry with peri-urban dairy establishments built in order to improve milk availability to rural communities. This study has been the first attempt to estimate the prevalence of brucellosis in Zimbabwe as a whole, using samples submitted to the Central veterinary laboratory between 2010 and 2014. A total of 156 farms were tested with Rose Bengal Test (RBT), Complement Fixation Test (CFT) and Milk Ring Test (MRT). Parallel testing was used to determine whether or not a farm was to be considered positive: 30.1% (95% C.I.: 23.5% - 37.8%) of the farms tested were found positive (47/156). Harare district had the highest number of sample submissions with 6 out of 17 farms testing positive (P = 37.5%; 95% C.I.: 18.4% - 61.7%). Awareness of milk-borne zoonoses is reportedly generally low in farmers (41.5%). This is even more the case in small-holder farmers who have higher likelihoods of selling or ingesting raw milk compared with dairy farmers. The results show the need to carry out surveillance of brucellosis in cattle in Zimbabwe to understand the spatial distribution of the disease in the country. This is particularly relevant given the zoonotic and economic implications of this disease.

Outbreak of unusual Salmonella enterica serovar Typhimurium monophasic variant 1,4 [5],12:i:-, Italy, June 2013 to September 2014.

Monophasic variant of Salmonella enterica subspecies enterica serovar Typhimurium (monophasic S. Typhimurium), with antigenic structure 1,4,[5],12:i:-, appears to be of increasing importance in Europe. In Italy, monophasic S. Typhimurium represented the third most frequent Salmonella serovar isolated from human cases between 2004 and 2008. From June 2013 to October 2014, a total of 206 human cases of salmonellosis were identified in Abruzzo region (Central Italy). Obtained clinical isolates characterised showed S. Typhimurium 1,4,[5],12:i:- with sole resistance to nalidixic acid, which had never been observed in Italy in monophasic S. Typhimurium, neither in humans nor in animals or foods. Epidemiological, microbiological and environmental investigations were conducted to try to identify the outbreak source. Cases were interviewed using a standardised questionnaire and microbiological tests were performed on human as well as environmental samples, including samples from fruit and vegetables, pigs, and surface water. Investigation results did not identify the final vehicle of human infection, although a link between the human cases and the contamination of irrigation water channels was suggested.

Draft Genome Sequences of 19 Salmonella enterica Serovar Typhimurium [4,5:i:-] Strains Resistant to Nalidixic Acid from a Long-Term Outbreak in Italy.

Here, we present the draft genome sequences of 19 Salmonella enterica serovar Typhimurium monophasic variant [4,5:i:-] strains involved in a long-term salmonellosis outbreak that occurred in central Italy in 2013 to 2014.

The 2011 West Nile disease outbreak in Sardinia region, Italy.

In 2011, strains of West Nile Virus (WNV) belonging to lineage 1 spread for the first time in Sardinia region (Italy). In contrast to previous WNV Italian incursion, the strains were found in Culex modestus and, more surprisingly, they were able to cause severe clinical signs in the affected birds. Based on the partial sequence of the NS3 encoding gene, the Sardinian WNV strains demonstrated a high similarity with the other WNV strains recently detected in the Mediterranean Basin. Nonetheless, the 2011 Sardinian sequences were grouped in a distinct sub-cluster. Both the NS3-249P and NS3-249T genotypes were detected in the Sardinian outbreaks confirming that the co-circulation of different genotypes in the affected population might be common for WNV as for many RNA viruses. No association, however, was observed between virulence and viral genotype.

Vector borne infections in Italy: results of the integrated surveillance system for West Nile disease in 2013.

The epidemiology of West Nile disease (WND) is influenced by multiple ecological factors and, therefore, integrated surveillance systems are needed for early detecting the infection and activating consequent control actions. As different animal species have different importance in the maintenance and in the spread of the infection, a multispecies surveillance approach is required. An integrated and comprehensive surveillance system is in place in Italy aiming at early detecting the virus introduction, monitoring the possible infection spread, and implementing preventive measures for human health. This paper describes the integrated surveillance system for WND in Italy, which incorporates data from veterinary and human side in order to evaluate the burden of infection in animals and humans and provide the public health authorities at regional and national levels with the information needed for a fine tune response.

Epidemiology of West Nile disease in Europe and in the Mediterranean Basin from 2009 to 2013.

West Nile virus (WNV) transmission has been confirmed in the last four years in Europe and in the Mediterranean Basin. An increasing concern towards West Nile disease (WND) has been observed due to the high number of human and animal cases reported in these areas confirming the importance of this zoonosis. A new epidemiological scenario is currently emerging: although new introductions of the virus from abroad are always possible, confirming the epidemiological role played by migratory birds, the infection endemisation in some European territories today is a reality supported by the constant reoccurrence of the same strains across years in the same geographical areas. Despite the WND reoccurrence in the Old World, the overwintering mechanisms are not well known, and the role of local resident birds or mosquitoes in this context is poorly understood. A recent new epidemiological scenario is the spread of lineage 2 strain across European and Mediterranean countries in regions where lineage 1 strain is still circulating creating favourable conditions for genetic reassortments and emergence of new strains. This paper summarizes the main epidemiological findings on WNV occurrence in Europe and in the Mediterranean Basin from 2009 to 2013, considering potential future spread patterns.

Systems for prevention and control of epidemic emergencies.

The development of early warning systems is fundamental for preventing the spread of infectious diseases. Data collection, however, is a costly activity and it is not possible to implement early warning systems everywhere and for all possible events. Hence, tools helping to improve the focus of surveillance efforts are of paramount importance. Risk assessment methods and other provisional modelling techniques may permit to estimate the probability of introduction and spread of infectious diseases in different geographical areas. Similarly, efficient information systems must be in place to assist the veterinary services in case of epidemic emergencies in order to support the prompt application of control measures for the containment of the infection and the reduction of the magnitude of negative consequences. This review describes two recent approaches to the estimation of the probability of introduction and spread of infectious diseases based on the use of risk maps/ spatial modelling and Social Network Analysis (SNA) techniques. The review also describes a web application developed in Italy to help official veterinary services to trace animals in case of outbreaks of infectious diseases.

Estimation of the sensitivity of the surveillance system for avian influenza in the western region of Cuba.

Although avian influenza (AI) virus of H5 and H7 subtypes has the potential to mutate to a highly pathogenic form and cause very high mortalities in some poultry species, most AI infections in poultry are due to low pathogenic AI (LPAI). Hence serological surveys, coupled with passive surveillance activities, are essential to detect sub-clinical infections by LPAI viruses, H5 and H7 subtypes. However the proper planning of an active surveillance system should be based on a careful estimation of its performance. Therefore, the sensitivity of the active surveillance system for AI in the western region of Cuba was assessed by a stochastic model quantifying the probability of revealing at least one animal infected by H5 or H7 subtype. The diagnostic sensitivity of the haemagglutination inhibition assay and different levels of within-flock prevalence (5%, 12% and 30%) were considered. The sensitivity of the surveillance system was then assessed under five different samples size scenarios: testing 20, 30, 40, 50 or 60 animals in each flock. Poultry flock sites in the western region of Cuba with a size ranging from 10,000 to 335,000 birds were included in the study.

Risk factors for the persistence of bovine brucellosis in Sicily from 2008 to 2010.

Bovine brucellosis is a worldwide zoonotic disease, responsible for economic and reproductive losses in farmed animals. Despite the application of control and eradication measures, after several years the brucellosis situation still remains problematic in Sicily, one of the regions with the highest prevalence of infection in Italy. The objective of this paper is to evaluate the possible risk factors influencing the presence of bovine brucellosis in Sicily. Data on existing cattle herds in Sicily and on their structure in 2008, 2009 and 2010 were extracted from the national database (NDb) of holdings and animals. Several possible brucellosis risk factors were considered, in particular herd-related factors and the performances of the control activities carried out by the veterinary services and recorded into national animal health information system (SANAN). Statistical analysis (χ(2) and Mann-Whitney tests) and spatial cluster analysis using a simple spatial Bernoulli model were performed. The multivariable logistic regression was also performed using SPSS for Windows. The spatial distribution of infected herds shows two main clusters of infection, in Messina and in Siracusa provinces. The results of the statistical analysis performed show that the infection is associated to the largest herds (P<0.0001) and to beef and mixed herds (P<0.0001). A significant association can be observed between the infected status in 2010 and presence of positive animals in 2008 or in 2009 (P<0.0001). This article shows the importance of detailed and reliable epidemiological data through information systems, allowing a continuous monitoring of the health status of animal populations and also the evaluation and re-planning of veterinary activities.

The national information system for the notification of animal diseases in Italy.

This paper describes the national system for the notification and management of outbreaks of animal diseases in Italy (Sistema Informativo Nazionale Malattie Animali: SIMAN). The main objective of the system is to provide a tool for the management of epidemic emergencies and to fulfil the information obligations towards international organisations, such as the European Commission and World Organisation for Animal Health (Office International des Épizooties: OIE). SIMAN was thus designed to collect all relevant information on outbreaks of animal diseases and to provide a useful tool for the management of activities to be implemented in emergencies. SIMAN is able to collect and report information concerning suspected or confirmed animal outbreaks in a consistent way and allows veterinary services to enter data electronically, instead of using paperwork, within the framework of the process of e-government and dematerialisation of the administrative acts. Data are immediately accessible for local and national authorities. The system provides the relevant national authorities with information relative to the planning of control measures in case epidemic emergencies. SIMAN is part of the e-government process that involves all public administrations of the European Union (EU) and refers to the use of information and communication technologies for the digital processing of documents so as to simplify the system and to make administrative procedures on the Internet much easier to follow.