27 results on '"Paola De Benedictis"'
Search Results
2. Alternative Methods to Current In Vivo Procedures to Address the 3Rs Tenet in Rabies Proficiency Testing
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Maira Zorzan, Morgane Gourlaouen, Stefania Leopardi, and Paola De Benedictis
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rabies ,proficiency testing ,3Rs tenet ,in vitro/ex vivo technologies ,Microbiology ,QR1-502 - Abstract
Canine rabies is responsible for an estimated 59,000 human deaths every year. In an attempt to reach the ZeroBy30 goal, robust disease surveillance coupled with improved diagnostics play a paramount role in ensuring reliable data and gradually attesting rabies control advancements. In this context, proficiency testing is organized to harmonize rabies diagnostic capacities. In most exercises, rabies-positive samples consist of brains collected from intracerebrally inoculated mice. This procedure causes distress and severe suffering to animals, raising important ethical concerns that can no longer be ignored. In the last decades, the 3Rs tenet (Replace, Reduce, Refine) has been successfully implemented in several scientific areas, and we strongly support its application in the framework of rabies proficiency testing. Here, we discuss cell-based technologies as innovative sustainable in vitro candidate systems to replace in vivo experiments for the production of proficiency testing samples. The application of these alternative methods can allow completely in vitro or ex vivo production of rabies proficiency testing panels, which would represent an important replacement or reduction/refinement for current in vivo procedures.
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- 2022
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3. The Importance of Accurate Host Species Identification in the Framework of Rabies Surveillance, Control and Elimination
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Paola De Benedictis, Stefania Leopardi, Wanda Markotter, and Andres Velasco-Villa
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wildlife reservoir ,host identification ,epidemiology ,rabies ,infectious diseases ,Microbiology ,QR1-502 - Abstract
Accurate host identification is paramount to understand disease epidemiology and to apply appropriate control measures. This is especially important for multi-host pathogens such as the rabies virus, a major and almost invariably fatal zoonosis that has mobilized unanimous engagement at an international level towards the final goal of zero human deaths due to canine rabies. Currently, diagnostic laboratories implement a standardized identification using taxonomic keys. However, this method is challenged by high and undiscovered biodiversity, decomposition of carcasses and subjective misevaluation, as has been attested to by findings from a cohort of 242 archived specimens collected across Sub-Saharan Africa and submitted for rabies diagnosis. We applied two simple and cheap methods targeting the Cytochrome b and Cytochrome c oxidase subunit I to confirm the initial classification. We therefore suggest prioritizing a standardized protocol that includes, as a first step, the implementation of taxonomic keys at a family or subfamily level, followed by the molecular characterization of the host species.
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- 2022
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4. A Three-Year Biocrime Sanitary Surveillance on Illegally Imported Companion Animals
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Monia Cocchi, Patrizia Danesi, Gabrita De Zan, Marta Leati, Laura Gagliazzo, Margherita Ruggeri, Manlio Palei, Alessandro Bremini, Marie-Christin Rossmann, Melanie Lippert-Petscharnig, Michael-Dieter Mansfeld, Silvia Deotto, Sofia Leardini, Federica Gobbo, Paolo Zucca, and Paola De Benedictis
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public health ,companion animals ,illegal trade ,zoonoses ,rabies ,Canine Parvovirus ,Medicine - Abstract
The illegal trade of companion animals in the European Union poses several legal, ethical and health issues to the entire community. In the framework of the Biocrime Interreg project between Italy and Austria, we surveyed puppies and kittens confiscated at the borders to identify the most frequent pathogens associated with (i) the risk of spread within the shelter, (ii) the development of fatal disease and (iii) the zoonotic potential. From January 2018 to December 2020, we examined a total of 613 puppies and 62 kittens coming from 44 requisitions. Feces, skin specimens and blood sera from confiscated animals were tested to verify the presence of major infections and to assess the rabies post-vaccination immunity. Out of the total of individuals under investigation, necropsies and laboratory investigations were also performed on 79 puppies and three kittens that had died during the observation period. Results indicated a high prevalence of Canine Parvovirus (CPV) and Giardia spp. infections, CPV as the most likely cause of fatal gastroenteritis in puppies and Salmonella and Microsporum canis as major zoonotic pathogens. Conversely, both extended spectrum beta lactamases Escherichia coli and methicillin resistant Staphylococcus pseudintermedius strains as rare findings. Results highlighted that illegal animal trade could expose the human population to potential zoonotic risk and naïve animal population to potentially disrupting epidemic waves, both of these issues being largely underestimated when buying companion animals.
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- 2021
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5. Development of broad‐spectrum human monoclonal antibodies for rabies post‐exposure prophylaxis
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Paola De Benedictis, Andrea Minola, Elena Rota Nodari, Roberta Aiello, Barbara Zecchin, Angela Salomoni, Mathilde Foglierini, Gloria Agatic, Fabrizia Vanzetta, Rachel Lavenir, Anthony Lepelletier, Emma Bentley, Robin Weiss, Giovanni Cattoli, Ilaria Capua, Federica Sallusto, Edward Wright, Antonio Lanzavecchia, Hervé Bourhy, and Davide Corti
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human monoclonal antibody ,lyssaviruses ,post‐exposure prophylaxis ,rabies ,Medicine (General) ,R5-920 ,Genetics ,QH426-470 - Abstract
Abstract Currently available rabies post‐exposure prophylaxis (PEP) for use in humans includes equine or human rabies immunoglobulins (RIG). The replacement of RIG with an equally or more potent and safer product is strongly encouraged due to the high costs and limited availability of existing RIG. In this study, we identified two broadly neutralizing human monoclonal antibodies that represent a valid and affordable alternative to RIG in rabies PEP. Memory B cells from four selected vaccinated donors were immortalized and monoclonal antibodies were tested for neutralizing activity and epitope specificity. Two antibodies, identified as RVC20 and RVC58 (binding to antigenic site I and III, respectively), were selected for their potency and broad‐spectrum reactivity. In vitro, RVC20 and RVC58 were able to neutralize all 35 rabies virus (RABV) and 25 non‐RABV lyssaviruses. They showed higher potency and breath compared to antibodies under clinical development (namely CR57, CR4098, and RAB1) and commercially available human RIG. In vivo, the RVC20–RVC58 cocktail protected Syrian hamsters from a lethal RABV challenge and did not affect the endogenous hamster post‐vaccination antibody response.
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- 2016
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6. Further Evidence of Inadequate Quality in Lateral Flow Devices Commercially Offered for the Diagnosis of Rabies
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Antonia Klein, Anna Fahrion, Stefan Finke, Marina Eyngor, Shiri Novak, Boris Yakobson, Ernest Ngoepe, Baby Phahladira, Claude Sabeta, Paola De Benedictis, Morgane Gourlaouen, Lillian A. Orciari, Pamela A. Yager, Crystal M. Gigante, M. Kimberly Knowles, Christine Fehlner-Gardiner, Alexandre Servat, Florence Cliquet, Denise Marston, Lorraine M. McElhinney, Trudy Johnson, Anthony R. Fooks, Thomas Müller, and Conrad M. Freuling
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rabies ,diagnostics ,lateral flow devices ,validation ,Medicine - Abstract
As a neglected zoonotic disease, rabies causes approximately 5.9 × 104 human deaths annually, primarily affecting low- and middle-income countries in Asia and Africa. In those regions, insufficient surveillance is hampering adequate medical intervention and is driving the vicious cycle of neglect. Where resources to provide laboratory disease confirmation are limited, there is a need for user-friendly and low-cost reliable diagnostic tools that do not rely on specialized laboratory facilities. Lateral flow devices (LFD) offer an alternative to conventional diagnostic methods and may strengthen control efforts in low-resource settings. Five different commercially available LFDs were compared in a multi-centered study with respect to their diagnostic sensitivity and their agreement with standard rabies diagnostic techniques. Our evaluation was conducted by several international reference laboratories using a broad panel of samples. The overall sensitivities ranged from 0% up to 62%, depending on the LFD manufacturer, with substantial variation between the different laboratories. Samples with high antigen content and high relative viral load tended to test positive more often in the Anigen/Bionote test, the latter being the one with the best performance. Still, the overall unsatisfactory findings corroborate a previous study and indicate a persistent lack of appropriate test validation and quality control. At present, the tested kits are not suitable for in-field use for rabies diagnosis, especially not for suspect animals where human contact has been identified, as an incorrect negative diagnosis may result in human casualties. This study points out the discrepancy between the enormous need for such a diagnostic tool on the one hand, and on the other hand, a number of already existing tests that are not yet ready for use.
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- 2020
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7. Unexpected rabies variant identified in kinkajou (Potos flavus), Mato Grosso, Brazil
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Gianpiero Zamperin, Andres Velasco-Villa, Risia Lopes Negreiros, Isabella Monne, Eunice da Conceição Souza, P. R. D. Rocha, Ernani Machado de Lima, Angela Salomoni, Alice Fusaro, Stefania Leopardi, Vera Lúcia Zafino, and Paola De Benedictis
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0301 basic medicine ,Veterinary medicine ,medicine.medical_specialty ,biology ,Epidemiology ,Public health ,030106 microbiology ,Immunology ,General Medicine ,Kinkajou ,medicine.disease ,Microbiology ,03 medical and health sciences ,030104 developmental biology ,Infectious Diseases ,Geography ,Virology ,biology.animal ,Drug Discovery ,medicine ,Parasitology ,Rabies - Abstract
A second case of a novel rabies variant described once in a capuchin monkey from Mato Grosso, Brazil, was discovered in a rabid wild kinkajou from the same region, indicating a public health risk f...
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- 2020
8. Complete Genome Sequences of Five Rabies Virus Strains Obtained from Domestic Carnivores in Liberia
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Stephanie Mauti, Garmie Voupawoe, Simon Bonas, Varney Kamara, Hervé Bourhy, Morgane Gourlaouen, Paola De Benedictis, Jakob Zinsstag, Laurent Dacheux, Lyssavirus, épidémiologie et neuropathologie - Lyssavirus Epidemiology and Neuropathology, Institut Pasteur [Paris] (IP)-Université Paris Cité (UPCité), Centre National de Référence de la Rage - National Reference Center Rabies (CNR), Centre collaborateur de l'OMS - Rage / World Health Organization Collaboration Centres - Rabies (CC-OMS / WHO-CC), Institut Pasteur [Paris] (IP)-Organisation Mondiale de la Santé / World Health Organization Office (OMS / WHO)-Université Paris Cité (UPCité), Swiss Tropical and Public Health Institute [Basel], University of Basel (Unibas), Leon Quist Ledlum Central Veterinary Laboratory, Ministry of Agriculture [Monrovia, Liberia], Istituto Zooprofilattico Sperimentale delle Venezie (IZSVe), and This work was funded through the Global Alliance for Vaccines and Immunisation (GAVI) (VISLACRV19122014, work stream 3), the Wolfermann Nägeli Foundation, the Swiss African Research Cooperation (SARECO), research funds from Stay on Track of the University of Basel, the Stiftung für wissenschaftliche Forschung (SWF), the Freiwillige Akademische Gesellschaft (FAG), and the Novartis Foundation for Biomedical Research.
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[SDV]Life Sciences [q-bio] ,Genome Sequences ,complete genome ,rabies ,lyssavirus ,Liberia ,phylogeny ,Immunology and Microbiology (miscellaneous) ,RABV ,Africa ,dog ,[SDV.MP.VIR]Life Sciences [q-bio]/Microbiology and Parasitology/Virology ,Genetics ,Molecular Biology - Abstract
International audience; Similar to other African countries, canine rabies is endemic in Liberia. However, data concerning the genetic diversity of rabies virus isolates circulating in this country remain limited. We report here the complete genome sequences of five rabies viruses obtained from domestic animals. All of them belonged to subgroup H within the Africa 2 clade.
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- 2022
9. Spillover of West Caucasian Bat Lyssavirus (WCBV) in a Domestic Cat and Westward Expansion in the Palearctic Region
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Fabio Parca, Paola De Benedictis, Giuseppe Manna, Pamela Priori, Petra Drzewnioková, Andrea Lombardo, Francesca Festa, Stefania Leopardi, Dino Scaravelli, Ettore Barneschi, Andrea Maroni Ponti, and Barbara Zecchin
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Male ,animal structures ,spillover ,Rabies ,bats ,Zoology ,animal–human encroachment ,Microbiology ,Article ,law.invention ,Russia ,West Caucasian Bat Lyssavirus ,law ,Virology ,Chiroptera ,Rhabdoviridae Infections ,Quarantine ,medicine ,Animals ,Humans ,Lyssavirus ,biology ,Miniopterus schreibersii ,biology.organism_classification ,medicine.disease ,QR1-502 ,Infectious Diseases ,Italy ,Epidemiological surveillance ,Cats ,Mammal ,Female ,Public Health ,Post-Exposure Prophylaxis - Abstract
In June 2020, a cat from Arezzo (Italy) that died from a neurological disease was diagnosed with West Caucasian Bat Lyssavirus (WCBV). The virus retained high identity across the whole-genome with the reference isolate found in 2002 from a Russian bent-winged bat. We applied control measures recommended by national regulations, investigated a possible interface between cats and bats using visual inspections, bioacoustics analyses and camera trapping and performed active and passive surveillance in bats to trace the source of infection. People that were exposed to the cat received full post-exposure prophylaxis while animals underwent six months of quarantine. One year later, they are all healthy. In a tunnel located near the cat’s house, we identified a group of bent-winged bats that showed virus-neutralizing antibodies to WCBV across four sampling occasions, but no virus in salivary swabs. Carcasses from other bat species were all negative. This description of WCBV in a non-flying mammal confirms that this virus can cause clinical rabies in the absence of preventive and therapeutic measures, and highlights the lack of international guidelines against divergent lyssaviruses. We detected bent-winged bats as the most probable source of infection, testifying the encroachment between these bats and pets/human in urban areas and confirming free-ranging cats as potential hazard for public health and conservation.
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- 2021
10. A Three-Year Biocrime Sanitary Surveillance on Illegally Imported Companion Animals
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Manlio Palei, Federica Gobbo, Michael-Dieter Mansfeld, Alessandro Bremini, Margherita Ruggeri, Paolo Zucca, Melanie Lippert-Petscharnig, Sofia Leardini, Patrizia Danesi, Monia Cocchi, Paola De Benedictis, Marta Leati, Gabrita De Zan, Marie-Christin Rossmann, Laura Gagliazzo, and Silvia Deotto
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illegal trade ,Microbiology (medical) ,medicine.medical_specialty ,Salmonella ,Population ,rabies ,Canine Parvovirus ,medicine.disease_cause ,Article ,Environmental health ,companion animals ,medicine ,public health ,zoonoses ,Giardia ,Immunology and Allergy ,media_common.cataloged_instance ,Microsporum canis ,European union ,education ,Molecular Biology ,Feces ,media_common ,education.field_of_study ,General Immunology and Microbiology ,biology ,business.industry ,Public health ,medicine.disease ,biology.organism_classification ,Infectious Diseases ,Medicine ,Rabies ,business - Abstract
The illegal trade of companion animals in the European Union poses several legal, ethical and health issues to the entire community. In the framework of the Biocrime Interreg project between Italy and Austria, we surveyed puppies and kittens confiscated at the borders to identify the most frequent pathogens associated with (i) the risk of spread within the shelter, (ii) the development of fatal disease and (iii) the zoonotic potential. From January 2018 to December 2020, we examined a total of 613 puppies and 62 kittens coming from 44 requisitions. Feces, skin specimens and blood sera from confiscated animals were tested to verify the presence of major infections and to assess the rabies post-vaccination immunity. Out of the total of individuals under investigation, necropsies and laboratory investigations were also performed on 79 puppies and three kittens that had died during the observation period. Results indicated a high prevalence of Canine Parvovirus (CPV) and Giardia spp. infections, CPV as the most likely cause of fatal gastroenteritis in puppies and Salmonella and Microsporum canis as major zoonotic pathogens. Conversely, both extended spectrum beta lactamases Escherichia coli and methicillin resistant Staphylococcus pseudintermedius strains as rare findings. Results highlighted that illegal animal trade could expose the human population to potential zoonotic risk and naïve animal population to potentially disrupting epidemic waves, both of these issues being largely underestimated when buying companion animals.
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- 2021
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11. Rabies control in Liberia: Joint efforts towards Zero by 30
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Baba Soumaré, Morgane Gourlaouen, Cristian De Battisti, Louis Hendrik Nel, Luis Filipe L de J Loureiro, Roland Varkpeh, Jakob Zinsstag, Garmie Voupawoe, Varney Kamara, Jesse D. Blanton, Andre Coetzer, Sonpon Sieh, Abdallah Traoré, Stefania Leopardi, Paola De Benedictis, Wolde Abebe, Laurent Dacheux, Terence Peter Scott, Monique Léchenne, Gwenaelle Dauphin, Hervé Bourhy, Simon Bonas, Angélique Angot, Stephanie Mauti, Ministry of Agriculture [Liberia], Swiss Tropical and Public Health Institute [Basel], University of Basel (Unibas), National Public Health Institute of Liberia (NPHIL), Laboratoire Central Vétérinaire [Bamako, Mali], Food and Agriculture Organization of the United Nations [Rome, Italie] (FAO), Universidade Lusofona Medicina Veterinaria, Food and Agriculture Organization of the United Nations [Accra, Ghana] (FAO), CEVA Santé Animale [Libourne, France] (Laboratoire Vétérinaire Pharmaceutique), Food and Agriculture Organization of the United Nations [Monrovia, Libéria] (FAO), University of Pretoria [South Africa], Global Alliance for Rabies Control [South Africa], Global Alliance for Rabies Control [Manhattan, Kansas], Centers for Disease Control and Prevention [Atlanta] (CDC), Centers for Disease Control and Prevention, Lyssavirus, épidémiologie et neuropathologie - Lyssavirus Epidemiology and Neuropathology, Institut Pasteur [Paris] (IP), FAO Reference Centre for Rabies, Istituto Zooprofilattico Sperimentale delle Venezie (IZSVe), Environment and Sustainability Institute [Penryn, UK], University of Exeter, This work was supported through the Global Alliance for Vaccines and Immunisation (GAVI) (VISLACRV19122014, Workstream 3), the Wolfermann Nägeli Foundation, the Swiss African Research Cooperation (SARECO) and research funds from Stay on Track of the University of Basel. FAO support was possible thanks to the GHSA project funded by USAID., We thank the dog owners, the County Livestock Officers, the Health Surveillance Officers, the members of the OH platform and the laboratory staff for their great commitment. We also want to acknowledge Lisa Crump for the language editing., and Institut Pasteur [Paris]
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0301 basic medicine ,Male ,Rabies ,Veterinary (miscellaneous) ,030231 tropical medicine ,Rabies lyssavirus ,Mass Vaccination ,Polymerase Chain Reaction ,03 medical and health sciences ,0302 clinical medicine ,Dogs ,[SDV.MHEP.MI]Life Sciences [q-bio]/Human health and pathology/Infectious diseases ,Environmental health ,Medicine ,Animals ,Humans ,National level ,Dog Diseases ,Diagnostic ,Phylogeny ,Disease surveillance ,[SDV.MHEP.ME]Life Sciences [q-bio]/Human health and pathology/Emerging diseases ,[SDV.BA.MVSA]Life Sciences [q-bio]/Animal biology/Veterinary medicine and animal Health ,business.industry ,Diagnostic Tests, Routine ,SARE tool ,030108 mycology & parasitology ,medicine.disease ,Liberia ,3. Good health ,Vaccination ,Phylogenetics ,Infectious Diseases ,One Health ,Molecular Diagnostic Techniques ,Rabies Vaccines ,GDREP tool ,Rabies virus ,Insect Science ,Action plan ,DNA, Viral ,[SDV.MP.VIR]Life Sciences [q-bio]/Microbiology and Parasitology/Virology ,Parasitology ,Female ,Rabies control ,business - Abstract
International audience; Despite declaration as a national priority disease, dog rabies remains endemic in Liberia, with surveillance systems and disease control activities still developing. The objective of these initial efforts was to establish animal rabies diagnostics, foster collaboration between all rabies control stakeholders, and develop a short-term action plan with estimated costs for rabies control and elimination in Liberia. Four rabies diagnostic tests, the direct fluorescent antibody (DFA) test, the direct immunohistochemical test (dRIT), the reverse transcriptase polymerase chain reaction (RT-PCR) assay and the rapid immunochromatographic diagnostic test (RIDT), were implemented at the Central Veterinary Laboratory (CVL) in Monrovia between July 2017 and February 2018. Seven samples (n=7) out of eight suspected animals were confirmed positive for rabies lyssavirus, and molecular analyses revealed that all isolates belonged to the Africa 2 lineage, subgroup H. During a comprehensive in-country One Health rabies stakeholder meeting in 2018, a practical workplan, a short-term action plan and an accurately costed mass dog vaccination strategy were developed. Liberia is currently at stage 1.5/5 of the Stepwise Approach towards Rabies Elimination (SARE) tool, which corresponds with countries that are scaling up local-level interventions (e.g. dog vaccination campaigns) to the national level. Overall an estimated 5.3 - 8 million USD invested over 13 years is needed to eliminate rabies in Liberia by 2030. Liberia still has a long road to become free from dog-rabies. However, the dialogue between all relevant stakeholders took place, and disease surveillance considerably improved through implementing rabies diagnosis at the CVL. The joint efforts of diverse national and international stakeholders laid important foundations to achieve the goal of zero dog-mediated human rabies deaths by 2030.
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- 2020
12. A combination of two human monoclonal antibodies cures symptomatic rabies
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Fabrizia Zatta, Roberta Aiello, Elise Brisebard, Grégory Jouvion, Guilherme D. Melo, Paola De Benedictis, Hervé Bourhy, Florence Larrous, Gabriel Lepousez, Davide Corti, Andrea Minola, Lauriane Kergoat, Carine Moigneu, Angela Salomoni, Camille Mazo, Florian Sonthonnax, Lyssavirus, épidémiologie et neuropathologie - Lyssavirus Epidemiology and Neuropathology, Institut Pasteur [Paris] (IP), Perception et Mémoire / Perception and Memory, Institut Pasteur [Paris] (IP)-Centre National de la Recherche Scientifique (CNRS), Neuropathologie expérimentale / Experimental neuropathology, Institut Pasteur [Paris] (IP)-Université Paris Cité (UPCité), Humabs BioMed SA, Istituto Zooprofilattico Sperimentale delle Venezie (IZSVe), ANR-16-IFEC-0006,ToRRENT,Towards a combined post-exposure prophylaxis and successful treatment of rabies in humans(2016), ANR-11-INBS-0006,FLI,France Life Imaging(2011), Institut Pasteur [Paris], Institut Pasteur [Paris]-Centre National de la Recherche Scientifique (CNRS), and Institut Pasteur [Paris]-Université de Paris (UP)
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0301 basic medicine ,Medicine (General) ,QH426-470 ,medicine.disease_cause ,Antibodies, Viral ,Mice ,0302 clinical medicine ,[SDV.MHEP.MI]Life Sciences [q-bio]/Human health and pathology/Infectious diseases ,Medicine ,Monoclonal antibody therapy ,biology ,Antibodies, Monoclonal ,Articles ,Microbiology, Virology & Host Pathogen Interaction ,neuroinfectious diseases ,3. Good health ,Molecular Medicine ,Antibody ,Post-Exposure Prophylaxis ,Viral load ,medicine.drug_class ,Rabies ,Immunology ,rabies virus Subject Categories Immunology ,Monoclonal antibody ,Microbiology ,Article ,03 medical and health sciences ,Dogs ,R5-920 ,Viral envelope ,neglected diseases ,Genetics ,Animals ,Humans ,rabies virus ,Lyssavirus ,business.industry ,monoclonal antibody therapy ,Rabies virus ,[SDV.IMM.IMM]Life Sciences [q-bio]/Immunology/Immunotherapy ,medicine.disease ,biology.organism_classification ,Virology ,Antibodies, Neutralizing ,Virology & Host Pathogen Interaction ,immunotherapeutics ,030104 developmental biology ,Rabies Vaccines ,biology.protein ,business ,030217 neurology & neurosurgery - Abstract
Rabies is a neglected disease caused by a neurotropic Lyssavirus, transmitted to humans predominantly by the bite of infected dogs. Rabies is preventable with vaccines or proper post‐exposure prophylaxis (PEP), but it still causes about 60,000 deaths every year. No cure exists after the onset of clinical signs, and the case‐fatality rate approaches 100% even with advanced supportive care. Here, we report that a combination of two potent neutralizing human monoclonal antibodies directed against the viral envelope glycoprotein cures symptomatic rabid mice. Treatment efficacy requires the concomitant administration of antibodies in the periphery and in the central nervous system through intracerebroventricular infusion. After such treatment, recovered mice presented good clinical condition, viral loads were undetectable, and the brain inflammatory profile was almost normal. Our findings provide the unprecedented proof of concept of an antibody‐based therapeutic approach for symptomatic rabies., Rabies is an invariably fatal disease after the rabies virus has invaded the central nervous system and clinical signs have been manifested. This study brings the proof‐of‐concept that a cocktail of human monoclonal antibodies (mAbs) can cure symptomatic rabies.
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- 2020
13. An inter-laboratory trial as a tool to increase rabies diagnostic capabilities of Sub-Saharan African Veterinary laboratories
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Baba Soumaré, Calogero Terregino, Marzia Mancin, Stefania Leopardi, Paola De Benedictis, Barbara Zecchin, Angélique Angot, Morgane Gourlaouen, Charles Bebay, Cristian De Battisti, and Francesca Ellero
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RNA viruses ,Central Nervous System ,0301 basic medicine ,Viral Diseases ,Veterinary medicine ,Research Facilities ,RC955-962 ,Artificial Gene Amplification and Extension ,Pathology and Laboratory Medicine ,medicine.disease_cause ,Polymerase Chain Reaction ,Nervous System ,Biosafety ,0302 clinical medicine ,Zoonoses ,Arctic medicine. Tropical medicine ,Medicine and Health Sciences ,Public and Occupational Health ,Dog Diseases ,Veterinary Diagnostics ,Vaccination and Immunization ,Test (assessment) ,Infectious Diseases ,Veterinary Diseases ,Medical Microbiology ,Viral Pathogens ,Viruses ,Pathogens ,Anatomy ,Public aspects of medicine ,RA1-1270 ,Research Laboratories ,Research Article ,Neglected Tropical Diseases ,Veterinary Medicine ,Rabies ,Concordance ,Immunology ,030231 tropical medicine ,MEDLINE ,Context (language use) ,Research and Analysis Methods ,Microbiology ,Rabies Virus ,03 medical and health sciences ,Dogs ,medicine ,Animals ,Humans ,Molecular Biology Techniques ,Molecular Biology ,Microbial Pathogens ,Africa South of the Sahara ,business.industry ,Rabies virus ,Organisms ,Public Health, Environmental and Occupational Health ,Biology and Life Sciences ,Reverse Transcriptase-Polymerase Chain Reaction ,Tropical Diseases ,medicine.disease ,030104 developmental biology ,Lyssavirus ,Veterinary Science ,Preventive Medicine ,Laboratories ,business ,Rabies in animals ,Government Laboratories - Abstract
To achieve the goal of eliminating dog-mediated human rabies deaths by 2030, many African countries have agreed to list rabies as a priority zoonotic disease and to undertake both short and long-term control programs. Within this context, reliable local diagnosis is essential for the success of field surveillance systems. However, a harmonized, sustainable and supportive diagnostic offer has yet to be achieved in the continent. We herewith describe the organization and outcome of a proficiency test (PT) for the post-mortem diagnosis of rabies in animals, involving thirteen veterinary laboratories and one public health laboratory in Africa. Participants were invited to assess both the performance of the Direct Fluorescent Antibody (DFA) test and of a conventional RT-PCR. From the submitted results, while thirteen laboratories proved to be able to test the samples through DFA test, eleven performed the RT-PCR method; ten applied both techniques. Of note, the number of laboratories able to apply rabies RT-PCR had increased from four to ten after the exercise. Importantly, results showed a higher proficiency in applying the molecular test compared to the DFA test (concordance, sensitivity and specificity: 98.2%, 96.97% and 100% for RT-PCR; 87.69%, 89.23% and 86.15% for DFA test), indicating the feasibility of molecular methods to diagnose animal pathogens in Africa. Another positive outcome of this approach was that negative and positive controls were made available for further in-house validation of new techniques; in addition, a detailed questionnaire was provided to collect useful and relevant information on the diagnostic procedures and biosafety measures applied at laboratory level., Author summary Although Africa has the highest per capita death rate from rabies, the incidence of canine rabies on the continent remains a matter that needs to be further investigated; in addition, the lack of accurate information impairs the establishment of long-term actions to progress towards a rabies free status. In this scenario, creating efficient diagnostic facilities is of utmost importance for eradicating the disease and implementing effective surveillance programs in endemic areas. With the final objective of eliminating dog-mediated human rabies globally by 2030, sample submission to African veterinary laboratories for rabies diagnostic testing is expected to increase. Furthermore, in compliance with ISO/IEC 17025:2017 standard, participating to proficiency testing (PT) exercises becomes mandatory not only to regularly assess the validity of the in-house protocols but also to harmonize the techniques. Thirteen veterinary laboratories and one public health laboratory in Africa were invited to take part in the exercise in response to an invitation from the Istituto Zooprofilattico Sperimentale delle Venezie (IZSVe), Italy, which hosts the FAO rabies Reference Center. This study presents the results obtained by applying the DFA test and the conventional RT-PCR method to the PT panel, which show that even if laboratories preferably apply the DFA test to detect the presence of rabies in animal samples, the overall concordance of results was higher through molecular testing. Importantly, the organization of the exercise proved to be a good opportunity to update the vaccine cover status of laboratory staff.
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- 2020
14. Further Evidence of Inadequate Quality in Lateral Flow Devices Commercially Offered for the Diagnosis of Rabies
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Pamela A. Yager, M. Kimberly Knowles, Marina Eyngor, Lorraine M. McElhinney, Alexandre Servat, Anna Fahrion, Stefan Finke, Anthony R. Fooks, Shiri Novak, Lillian A. Orciari, Boris Yakobson, Trudy Johnson, Christine Fehlner-Gardiner, Crystal M. Gigante, Baby Phahladira, Morgane Gourlaouen, Claude T. Sabeta, Paola De Benedictis, Ernest Ngoepe, Florence Cliquet, Thomas Müller, Conrad M. Freuling, Antonia Klein, Denise A. Marston, Friedrich-Loeffler-Institut (FLI), Kimron Veterinary Institute (KVI), Onderstepoort Veterinary Institute (ARC - OVI), Istituto Zooprofilattico Sperimentale delle Venezie (IZSVe), Centers for Disease Control and Prevention [Atlanta] (CDC), Centers for Disease Control and Prevention, Canadian Food Inspection Agency, National Centre for Foreign Animal Disease, Laboratoire de la rage et de la faune sauvage de Nancy (LRFSN), Agence nationale de sécurité sanitaire de l'alimentation, de l'environnement et du travail (ANSES), and Animal and Plant Health Agency [Weybridge] (APHA)
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0301 basic medicine ,medicine.medical_specialty ,Diagnostic methods ,media_common.quotation_subject ,030231 tropical medicine ,lcsh:Medicine ,rabies ,Diagnostic tools ,Zoonotic disease ,Neglect ,03 medical and health sciences ,0302 clinical medicine ,[SDV.MHEP.MI]Life Sciences [q-bio]/Human health and pathology/Infectious diseases ,medicine ,diagnostics ,Quality (business) ,Intensive care medicine ,lateral flow devices ,media_common ,validation ,General Immunology and Microbiology ,business.industry ,Communication ,lcsh:R ,Public Health, Environmental and Occupational Health ,medicine.disease ,3. Good health ,Test (assessment) ,030104 developmental biology ,Infectious Diseases ,Rabies ,[SDV.SPEE]Life Sciences [q-bio]/Santé publique et épidémiologie ,business - Abstract
International audience; As a neglected zoonotic disease, rabies causes approximately 5.9 × 104 human deaths annually, primarily affecting low- and middle-income countries in Asia and Africa. In those regions, insufficient surveillance is hampering adequate medical intervention and is driving the vicious cycle of neglect. Where resources to provide laboratory disease confirmation are limited, there is a need for user-friendly and low-cost reliable diagnostic tools that do not rely on specialized laboratory facilities. Lateral flow devices (LFD) offer an alternative to conventional diagnostic methods and may strengthen control efforts in low-resource settings. Five different commercially available LFDs were compared in a multi-centered study with respect to their diagnostic sensitivity and their agreement with standard rabies diagnostic techniques. Our evaluation was conducted by several international reference laboratories using a broad panel of samples. The overall sensitivities ranged from 0% up to 62%, depending on the LFD manufacturer, with substantial variation between the different laboratories. Samples with high antigen content and high relative viral load tended to test positive more often in the Anigen/Bionote test, the latter being the one with the best performance. Still, the overall unsatisfactory findings corroborate a previous study and indicate a persistent lack of appropriate test validation and quality control. At present, the tested kits are not suitable for in-field use for rabies diagnosis, especially not for suspect animals where human contact has been identified, as an incorrect negative diagnosis may result in human casualties. This study points out the discrepancy between the enormous need for such a diagnostic tool on the one hand, and on the other hand, a number of already existing tests that are not yet ready for use.
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- 2020
15. Field postmortem rabies rapid immunochromatographic diagnostic test for resource-limited settings with further molecular applications
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Laurent Dacheux, Jakob Zinsstag, Hervé Bourhy, Paola De Benedictis, Pascal Cozette, Ibrahima Dicko, Enos Madaye, Pati Patient Pyana, Céline Mbilo, Morgane Gourlaouen, Emmanuel Couacy-Hymann, Casimir Kouakou, Vessaly Kallo, Abdallah Traoré, Service Naïssengar, Monique Léchenne, Stephanie Mauti, Lyssavirus, épidémiologie et neuropathologie - Lyssavirus Epidemiology and Neuropathology, Institut Pasteur [Paris] (IP), Environment and Sustainability Institute [Penryn, UK], University of Exeter, Institut de Recherche en Elevage pour le Developpement [N'Djamena, Tchad] (IRED), Laboratoire Central Vétérinaire [Bamako, Mali], Ecole Inter-États des Sciences et Médecine Vétérinaires de Dakar (EISMV), Direction des Services Vétérinaires, Laboratoire National d'Appui au Développement Agricole (LANADA), Laboratoire Central Vétérinaire de Bingerville, FAO Reference Centre for Rabies, Istituto Zooprofilattico Sperimentale delle Venezie (IZSVe), Swiss Tropical and Public Health Institute [Basel], University of Basel (Unibas), Institut National de Recherche Biomédicale [Kinshasa] (INRB), This work was supported through the Global Alliance for Vaccines and Immunisation (GAVI), the Wolfermann Nägeli Foundation, the Swiss African Research Cooperation (SARECO), the SWF Stiftung für wissenschaftliche Forschung, the Freiwillige Akademische Gesellschaft (FAG) Basel, the Bilateral Science and Technology Cooperation Programme of Switzerland with Asia and the Novartis Foundation for biomedical research., and Institut Pasteur [Paris]
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0301 basic medicine ,Rabies ,MESH: Diagnosis ,General Chemical Engineering ,030231 tropical medicine ,medicine.disease_cause ,General Biochemistry, Genetics and Molecular Biology ,03 medical and health sciences ,0302 clinical medicine ,MESH: Rabies ,Diagnosis ,medicine ,Animals ,Sampling (medicine) ,MESH: Animals ,Direct fluorescent antibody ,MESH: Diagnostic Tests, Routine ,Protocol (science) ,Postmortem Diagnosis ,Immunoassay ,medicine.diagnostic_test ,General Immunology and Microbiology ,business.industry ,Diagnostic Tests, Routine ,Brain biopsy ,General Neuroscience ,Rabies virus ,Gold standard (test) ,medicine.disease ,Virology ,3. Good health ,MESH: Rabies virus ,030104 developmental biology ,[SDV.MP.VIR]Life Sciences [q-bio]/Microbiology and Parasitology/Virology ,business ,MESH: Immunoassay - Abstract
International audience; Functional rabies surveillance systems are crucial to provide reliable data and increase the political commitment necessary for disease control. To date, animals suspected as rabies-positive must be submitted to a postmortem confirmation using classical or molecular laboratory methods. However, most endemic areas are in low-and middle-income countries where animal rabies diagnosis is restricted to central veterinary laboratories. Poor availability of surveillance infrastructure leads to serious disease underreporting from remote areas. Several diagnostic protocols requiring low technical expertise have been recently developed, providing opportunity to establish rabies diagnosis in decentralized laboratories. We present here a complete protocol for field postmortem diagnosis of animal rabies using a rapid immunochromatographic diagnostic test (RIDT), from brain biopsy sampling to the final interpretation. We complete the protocol by describing a further use of the device for molecular analysis and viral genotyping. RIDT easily detects rabies virus and other lyssaviruses in brain samples. The principle of such tests is simple: brain material is applied on a test strip where gold conjugated antibodies bind specifically to rabies antigens. The antigen-antibody complexes bind further to fixed antibodies on the test line, resulting in a clearly visible purple line. The virus is inactivated in the test strip, but viral RNA can be subsequently extracted. This allows the test strip, rather than the infectious brain sample, to be safely and easily sent to an equipped laboratory for confirmation and molecular typing. Based on a modification of the manufacturer's protocol, we found increased test sensitivity, reaching 98% compared to the gold standard June 2020 • 160 • e60008 • Page 2 of 29 reference method, the direct immunofluorescence antibody test. The advantages of the test are numerous: rapid, easy-to-use, low cost and no requirement for laboratory infrastructure, such as microscopy or cold-chain compliance. RIDTs represent a useful alternative for areas where reference diagnostic methods are not available.
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- 2020
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16. Dog rabies control in West and Central Africa: A review
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Pati Patient Pyana, Corneille Gnanvi, Morou Mounkaila, Andrée Prisca Ndjoug Ndour, Jakob Zinsstag, Richard Suu-Ire, Issaka Tiembré, Mathilde Tetchi, Céline Mbilo, Richard H. Lokossou, Garmie Voupawoe, Charles Bebay, Angélique Angot, Vessaly Kallo, Louis Hendrik Nel, Serge Alain Sadeuh-Mba, Paola De Benedictis, Abdallah Traoré, Cristóvão Manjuba, Roland Suluku, Mathurin Cyrille Tejiokem, Rakiswendé Constant Roamba, Guy Anicet Rerambyath, Pidemnéwé Pato, Beatrice Mouille, Bernardo Cassamá, Etienne Mokondjimobe, Moina Hasni Ebou, Andre Coetzer, Babasola O Olugasa, and Bassirou Bonfoh
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0301 basic medicine ,Rabies ,Veterinary (miscellaneous) ,030231 tropical medicine ,Population ,Notifiable disease ,03 medical and health sciences ,0302 clinical medicine ,Rabies vaccine ,Dogs ,medicine ,Animals ,Africa, Central ,Dog Diseases ,Socioeconomics ,education ,education.field_of_study ,Zoonosis ,030108 mycology & parasitology ,medicine.disease ,Human development (humanity) ,Infectious Diseases ,One Health ,Geography ,Rabies Vaccines ,Insect Science ,Parasitology ,Rural area ,Post-Exposure Prophylaxis ,medicine.drug - Abstract
Rabies is a neglected but preventable zoonotic disease that predominantly affects the most vulnerable populations living in remote rural areas of resource-limited countries. To date, every country on the African mainland is considered endemic for dog-mediated rabies with an estimated 21'500 human rabies deaths occurring each year. In 2018, the United Against Rabies collaboration launched the Global Strategic Plan to end human deaths from dog-mediated rabies by 2030. The epidemiology of rabies from most Western and Central African countries remains poorly defined, making it difficult to assess the overall rabies situation and progress towards the 2030 goal. In this review, we attempt to provide an overview of the current rabies situation in 22 West and Central African countries based on published scientific literature and information obtained from rabies focal points. To this end, information was collected on i) established surveillance, ii) diagnostic capacity, iii) post-exposure prophylaxis (PEP) availability and coverage, iv) dog population estimates, v) dog vaccination campaigns, vi) animal and human health communication (One Health), vii) molecular studies, viii) Knowledge, Attitude and Practices (KAP), ix) cost estimates and x) national control strategies. Although rabies is a notifiable disease in the majority of the studied countries, national surveillance systems do not adequately capture the disease. A general lack of rabies diagnostic capacity has an additional negative impact on rabies surveillance and attempts to estimate rabies burden. Recurrent shortages of human rabies vaccine are reported by all of the countries, with vaccine availability usually limited to major urban centers but no country has yet adopted the new WHO-recommended 1-week intradermal vaccination regimen. Most countries carry out subsidized mass dog vaccination campaigns on World Rabies Day. Such activities are indispensable to keep rabies in the public consciousness but are not of the scale and intensity that is required to eliminate rabies from the dog population. Countries will need to scale up the intensity of their campaigns, if they are to progress towards the 2030 goal. But more than half of the countries do not yet have reliable figures on their dog populations. Only two countries reached stage 2 on the Stepwise Approach towards Rabies Elimination ladder - indicating that their national governments have truly prioritized rabies elimination and are thus providing the necessary support and political buy-in required to achieve success. In summary, the sub-region of West and Central Africa seems to be divided into countries which have accepted the challenge to eliminate rabies with governments committed to pushing forward rabies elimination, while other countries have achieved some progress, but elimination efforts remain stuck due to lacking government commitment and financial constraints. The possibility to meet the 2030 goal without international solidarity is low, because more than two-thirds of the countries rank in the low human development group (HDI ≤ 152). Leading countries should act as role models, sharing their experiences and capacities so that no country is left behind. Unified and with international support it is possible to reach the common goal of zero human rabies deaths by 2030.
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- 2019
17. Disinfection protocols for necropsy equipment in rabies laboratories: Safety of personnel and diagnostic outcome
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Paola De Benedictis, Giovanni Cattoli, Roberta Aiello, Silvia Tiozzo Caenazzo, and Barbara Zecchin
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0301 basic medicine ,medicine.medical_specialty ,Rabies ,040301 veterinary sciences ,medicine.disease_cause ,Specimen Handling ,0403 veterinary science ,Toxicology ,03 medical and health sciences ,Virology ,Medical Laboratory Personnel ,medicine ,Humans ,Post-mortem diagnosis ,Intensive care medicine ,Antigens, Viral ,Direct fluorescent antibody ,Protocol (science) ,business.industry ,Rabies virus ,Routine laboratory ,Diagnostic test ,04 agricultural and veterinary sciences ,Gold standard (test) ,medicine.disease ,Molecular methods ,3. Good health ,Disinfection ,030104 developmental biology ,Molecular Diagnostic Techniques ,Equipment Contamination ,Autopsy ,Safety ,business ,Disinfectants - Abstract
In the last decades, molecular techniques have gradually been adopted for the rapid confirmation of results obtained through gold standard methods. However, international organisations discourage their use in routine laboratory investigations for rabies post-mortem diagnosis, as they may lead to false positive results due to cross-contamination. Cleaning and disinfection are essential to prevent cross-contamination of samples in the laboratory environment. The present study evaluated the efficacy of selected disinfectants on rabies-contaminated necropsy equipment under organic challenge using a carrier-based test. The occurrence of detectable Rabies virus (RABV) antigen, viable virus and RNA was assessed through the gold standard Fluorescent Antibody Test, the Rabies Tissue Culture Infection Test and molecular techniques, respectively. None of the tested disinfectants proved to be effective under label conditions. Off label disinfection protocols were found effective for oxidizing agents and phenolic, only. Biguanide and quaternary ammonium compound were both ineffective under all tested conditions. Overall, discordant results were obtained when different diagnostic tests were compared, which means that in the presence of organic contamination common disinfectants may not be effective enough on viable RABV or RNA. Our results indicate that an effective disinfection protocol should be carefully validated to guarantee staff safety and reliability of results.
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- 2016
18. Genetic and spatial characterization of the red fox (Vulpes vulpes) population in the area stretching between the Eastern and Dinaric Alps and its relationship with rabies and canine distemper dynamics
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Marco De Nardi, Luca Bargelloni, Adelaide Milani, Peter Hostnik, Zoltán Bagó, B. Crestanello, Tomislav Bedeković, Massimiliano Babbucci, Giovanni Cattoli, Federica Obber, Paola De Benedictis, C.V. Citterio, Bianca Zecchin, Christl A. Donnelly, Cristiano Vernesi, Monica Lorenzetto, Pierre Nouvellet, National Institute for Health Research, and Medical Research Council (MRC)
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Male ,RNA viruses ,0106 biological sciences ,0301 basic medicine ,Viral Diseases ,European People ,Vulpes ,Slovenia ,Foxes ,Population genetics ,Pathology and Laboratory Medicine ,01 natural sciences ,Geographical locations ,Gene flow ,Settore BIO/05 - ZOOLOGIA ,Zoonoses ,Prevalence ,Medicine and Health Sciences ,Ethnicities ,Mammals ,education.field_of_study ,Multidisciplinary ,Eukaryota ,Italian People ,3. Good health ,Europe ,Infectious Diseases ,Italy ,Medical Microbiology ,Austria ,Viral Pathogens ,Viruses ,Vertebrates ,Genetic structure ,Medicine ,Female ,Pathogens ,geographic locations ,Research Article ,Neglected Tropical Diseases ,Croatia ,Rabies ,General Science & Technology ,Science ,Population ,Zoology ,Biology ,Microbiology ,010603 evolutionary biology ,Rabies Virus ,03 medical and health sciences ,Dogs ,parasitic diseases ,MD Multidisciplinary ,Genetics ,medicine ,Animals ,European Union ,Distemper ,education ,Microbial Pathogens ,Evolutionary Biology ,Biology and life sciences ,Population Biology ,Canine distemper ,Organisms ,Tropical Diseases ,medicine.disease ,biology.organism_classification ,030104 developmental biology ,Infectious disease (medical specialty) ,Amniotes ,Genetics of Disease ,Biological dispersal ,Lyssavirus ,Population Groupings ,People and places ,Population Genetics ,Microsatellite Repeats - Abstract
Information on the population dynamics of a reservoir species have been increasingly adopted to understand and eventually predict the dispersal patterns of infectious diseases throughout an area. Although potentially relevant, to date there are no studies which have investigated the genetic structure of the red fox population in relation to infectious disease dynamics. Therefore, we genetically and spatially characterised the red fox population in the area stretching between the Eastern and Dinaric Alps, which has been affected by both distemper and rabies at different time intervals. Red foxes collected from north-eastern Italy, Austria, Slovenia and Croatia between 2006-2012, were studied using a set of 21 microsatellite markers. We confirmed a weak genetic differentiation within the fox population using Bayesian clustering analyses, and we were able to differentiate the fox population into geographically segregated groups. Our finding might be due to the presence of geographical barriers that have likely influenced the distribution of the fox population, limiting in turn gene flow and spread of infectious diseases. Focusing on the Italian red fox population, we observed interesting variations in the prevalence of both diseases among distinct fox clusters, with the previously identified Italy 1 and Italy 2 rabies as well as distemper viruses preferentially affecting different sub-groups identified in the study. Knowledge of the regional-scale population structure can improve understanding of the epidemiology and spread of diseases. Our study paves the way for an integrated approach for disease control coupling pathogen, host and environmental data to inform targeted control programs in the future.
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- 2019
19. The introduction of fox rabies into Italy (2008-2011) was due to two viral genetic groups with distinct phylogeographic patterns
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Ilaria Capua, Matteo Trolese, Angela Salomoni, Franco Mutinelli, Philippe Lemey, Giovanni Cattoli, Paola De Benedictis, Isabella Monne, Alice Fusaro, Nicola Ferrè, Angélique Angot, and Edward C. Holmes
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Microbiology (medical) ,Genes, Viral ,Rabies ,Molecular Sequence Data ,030231 tropical medicine ,Wildlife ,Foxes ,Biology ,Disease cluster ,Microbiology ,03 medical and health sciences ,0302 clinical medicine ,Intergenic region ,Amino acid mutation ,Phylogenetics ,Genetics ,medicine ,Animals ,Molecular Biology ,Phylogeny ,Ecology, Evolution, Behavior and Systematics ,030304 developmental biology ,0303 health sciences ,Phylogenetic tree ,medicine.disease ,Virology ,3. Good health ,Phylogeography ,Infectious Diseases ,Italy ,Rabies virus ,Evolutionary biology - Abstract
Fox rabies re-emerged in north-eastern Italy at the end of 2008 and circulated until early 2011. As with previous rabies epidemics, the Italian cases were linked to the epidemiological situation in adjacent regions. To obtain a comprehensive picture of the dynamics of the recent Italian epidemic, we performed a detailed evolutionary analysis of RABVs circulating in north-eastern Italy. Sequences were obtained for the hyper-variable region of the nucleoprotein gene, the complete glycoprotein gene, and the intergenic region G-L from 113 selected fox rabies cases. We identified two viral genetic groups, here referred to as Italy-1 and Italy-2. Phylogenetic and phylogeographic analyses revealed that both groups had been circulating in the Western Balkans and Slovenia in previous years and were only later introduced into Italy (into the Friuli Venezia Giulia region-FVG), occupying different areas of the Italian territories. Notably, viruses belonging to the Italy-1 group remained confined to the region of introduction and their spread was minimised by the implementation of oral fox vaccination campaigns. In contrast, Italy-2 viruses spread westward over a territory of 100 km from their first identification in FVG, likely crossing the northern territories where surveillance was inadequate. A genetic sub-group (Italy-2A), characterised by a unique amino acid mutation (D106A) in the N gene, was also observed to occupy a distinct geographic cluster. This molecular epidemiological analysis of the 2008–2011 fox rabies epidemic will contribute to future control programmes both at national and regional levels. In particular, our findings highlight the weaknesses of the national surveillance strategy in the period preceding rabies re-emergence, and of control plans implemented immediately after rabies notification, and underline the need of a coordinated approach at the regional level for both the surveillance and control of wildlife rabies.
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- 2013
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20. Evaluation of Six Commercially Available Rapid Immunochromatographic Tests for the Diagnosis of Rabies in Brain Material
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Ernest Ngoepe, Elisa Eggerbauer, Kore Schlottau, Thomas C. Mettenleiter, Bernd Hoffmann, Thomas Müller, Conrad M. Freuling, Claude T. Sabeta, and Paola De Benedictis
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RNA viruses ,0301 basic medicine ,Viral Diseases ,Veterinary medicine ,Artificial Gene Amplification and Extension ,Pathology and Laboratory Medicine ,medicine.disease_cause ,Biochemistry ,Polymerase Chain Reaction ,Chromatography, Affinity ,Zoonotic disease ,Fats ,0302 clinical medicine ,Zoonoses ,Medicine and Health Sciences ,Mammals ,biology ,lcsh:Public aspects of medicine ,Brain ,Genomics ,Viral Load ,Lipids ,Infectious Diseases ,Medical Microbiology ,Viral Pathogens ,Viruses ,Vertebrates ,RNA, Viral ,Pathogens ,Research Article ,Neglected Tropical Diseases ,lcsh:Arctic medicine. Tropical medicine ,Disease detection ,Rabies ,lcsh:RC955-962 ,Sample (material) ,030231 tropical medicine ,Research and Analysis Methods ,Microbiology ,Sensitivity and Specificity ,Rabies Virus ,03 medical and health sciences ,Dogs ,Extraction techniques ,Genetics ,medicine ,Animals ,Humans ,Immunochromatographic Assays ,Molecular Biology Techniques ,Microbial Pathogens ,Molecular Biology ,Lyssavirus ,Rabies virus ,Organisms ,Public Health, Environmental and Occupational Health ,Biology and Life Sciences ,lcsh:RA1-1270 ,Reverse Transcriptase-Polymerase Chain Reaction ,Tropical Diseases ,medicine.disease ,biology.organism_classification ,Virology ,RNA extraction ,030104 developmental biology ,Amniotes ,Reagent Kits, Diagnostic - Abstract
Rabies is a neglected zoonotic disease that causes an estimated 60,000 human deaths annually. The main burden lies on developing countries in Asia and Africa, where surveillance and disease detection is hampered by absence of adequate laboratory facilities and/or the difficulties of submitting samples from remote areas to laboratories. Under these conditions, easy-to-use tests such as immunochromatographic assays, i.e. lateral flow devices (LFD), may increase surveillance and improve control efforts. Several LFDs for rabies diagnosis are available but, except for one, there are no data regarding their performance. Therefore, we compared six commercially available LFDs for diagnostic and analytical sensitivity, as well as their specificity and their diagnostic agreement with standard rabies diagnostic techniques using different sample sets, including experimentally infected animals and several sets of field samples. Using field samples the sensitivities ranged between 0% up to 100% depending on the LFD and the samples, while for experimentally infected animals the maximum sensitivity was 32%. Positive results in LFD could be further validated using RT-qPCR and sequencing. In summary, in our study none of the tests investigated proved to be satisfactory, although the results somewhat contradict previous studies, indicating batch to batch variation. The high number of false negative results reiterates the necessity to perform a proper test validation before being marketed and used in the field. In this respect, marketing authorization and batch release control could secure a sufficient quality for these alternative tests, which could then fulfil their potential., Author Summary Despite being preventable with adequate biologicals, rabies still causes an estimated 60,000 human deaths annually. The main burden lies on developing countries in Asia and Africa, where dog rabies surveillance is hampered by laboratory confirmation of disease due to a number of reasons, including laboratory infrastructure and logistics. Lateral flow devices (LFD) may increase surveillance and improve control efforts. Several LFDs for rabies diagnosis are available but, except for one, there are no data available regarding their performance. Therefore, we compared six commercially available LFDs for diagnostic and analytical sensitivity. With sensitivities ranging from 0% up to 100% depending on the LFD and the samples, none of the tests investigated proved to be satisfactory, and the results somewhat contradict previous studies, indicating batch to batch variation. The high number of false negative results reiterates the necessity to perform a proper test validation before being marketed and used in the field. Only when sufficient quality is assured for these alternative tests, then they can fulfil their potential. In this respect, we demonstrated that positive results in LFD can be further validated and characterized using RT-qPCR and sequencing.
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- 2016
21. Serological methods used for rabies post vaccination surveys: An analysis
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Giovanni Cattoli, Calogero Terrregino, Ilaria Capua, Marzia Mancin, and Paola De Benedictis
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European level ,Foxes ,Enzyme-Linked Immunosorbent Assay ,Antibodies, Viral ,Diagnostic tools ,Rabies vaccination ,Serology ,Neutralization Tests ,Post vaccination ,medicine ,Animals ,Serologic Tests ,General Veterinary ,General Immunology and Microbiology ,business.industry ,Public Health, Environmental and Occupational Health ,Gold standard (test) ,medicine.disease ,Virology ,Vaccination ,Infectious Diseases ,Italy ,Rabies Vaccines ,Molecular Medicine ,Rabies ,business - Abstract
The assessment of fox immunity following oral rabies vaccination (ORV) is commonly applied to assess the efficacy of an ORV campaign in the field. Several ELISA kits have been developed and validated for their use for rabies serology in wildlife as an alternative to neutralizing techniques (NT), such as the fluorescent antibody virus neutralization test (FAVN) and the rapid fluorescent foci inhibition test (RFFIT). At a European level, NT and ELISA tests are used interchangeably and on different types of samples collected for vaccination follow-up. This has resulted in a difficulty in comparing the results generated with different diagnostic tools. We have evaluated (a) the effect of two different matrices commonly used for serology in red foxes on the results of the FAVN and (b) the performance of two commercially available ELISAs in comparison with the FAVN, as a gold standard, using a panel of over 700 field fox samples. Moderate agreement was observed when comparing results from different matrices. We found a very low level of agreement and low values of relative sensitivity and specificity of the ELISAs tested in comparison with the FAVN. Our findings confirm, using a vast collection of field samples obtained during post-vaccination surveillance campaigns in Italy, the need for improved reliability of certain serological tests.
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- 2012
22. Evolutionary history and dynamics of dog rabies virus in western and central Africa
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Ousmane Faye, William G. Dundon, Chiraz Talbi, Oumou Sangare, Amadou A. Sall, Emmanuel Nakouné, Edward C. Holmes, Djibo Gamatié, Paola De Benedictis, Edgard Valery Adjogoua, Ilaria Capua, Adama Sow, Abass Diarra, Bezeid Ould Elmamy, and Hervé Bourhy
- Subjects
Time Factors ,Rabies ,Lineage (evolution) ,Molecular Sequence Data ,Population ,medicine.disease_cause ,Coalescent theory ,Evolution, Molecular ,Dogs ,Species Specificity ,Viral Envelope Proteins ,Phylogenetics ,Virology ,medicine ,Animals ,Africa, Central ,Dog Diseases ,education ,Rabies transmission ,Antigens, Viral ,Lyssavirus ,Phylogeny ,Glycoproteins ,education.field_of_study ,biology ,Rabies virus ,Genetic Variation ,Sequence Analysis, DNA ,Nucleocapsid Proteins ,medicine.disease ,biology.organism_classification ,Africa, Western - Abstract
The burden of rabies in Africa is estimated at 24 000 human deaths year−1, almost all of which result from infection with dog rabies viruses (RABV). To investigate the evolutionary dynamics of RABV in western and central Africa, 92 isolates sampled from 27 African countries over 29 years were collected and sequenced. This revealed that RABV currently circulating in dogs in this region fell into a single lineage designated ‘Africa 2’. A detailed analysis of the phylogeographical structure of this Africa 2 lineage revealed strong population subdivision at the country level, with only limited movement of virus among localities, including a possible east-to-west spread across Africa. In addition, Bayesian coalescent analysis suggested that the Africa 2 lineage was introduced into this region of Africa only recently (probably
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- 2009
23. Pyrosequencing of the rabies virus glycoprotein gene to demonstrate absence of vaccine-associated rabies cases following oral vaccination
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Franco Mutinelli, Sabrina Marciano, Giovanni Cattoli, Paola De Benedictis, Cristian De Battisti, and Ilaria Capua
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Rabies ,Administration, Oral ,Foxes ,Biology ,Global Health ,medicine.disease_cause ,Viral Proteins ,Vaccine strain ,parasitic diseases ,medicine ,Animals ,Gene ,Glycoproteins ,Duck embryo vaccine ,chemistry.chemical_classification ,General Veterinary ,Vaccination ,Rabies virus ,medicine.disease ,Virology ,Peptide Fragments ,Rabies Vaccines ,chemistry ,Immunology ,Pyrosequencing ,Animal Science and Zoology ,Glycoprotein - Abstract
Replication competent vaccines have been used successfully for the control of terrestrial rabies, mainly in wildlife; however, these vaccine strains occasionally may induce rabies. In this study, a pyrosequencing protocol for the rapid identification of vaccine-associated rabies viruses was applied to the 2008-2011 Italian epidemic. There was no evidence of vaccine-associated rabies cases following oral vaccination of foxes with the SAG2 and SADB19 vaccine strains.
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- 2013
24. Genetic Characterization via Pyrosequencing
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Cristian De Battisti and Paola De Benedictis
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Sanger sequencing ,Biology ,medicine.disease ,biology.organism_classification ,Virology ,Hypervariable region ,Nucleoprotein ,symbols.namesake ,symbols ,medicine ,Pyrosequencing ,Rabies ,Typing ,Gene ,Lyssavirus - Abstract
Pyrosequencing is an alternative sequencing technique of small DNA fragments based on the sequencing-by-synthesis principle. Compared to the Sanger method, pyrosequencing is less costly and time consuming, although the applications of this technique do not completely overlap with those of the conventional method. In the last decade, pyrosequencing technology has been applied successfully to the typing of a number of pathogens, including lyssaviruses. An efficient protocol to lyssavirus detection and typing, by targeting the hypervariable region of the rabies nucleoprotein (N) gene, has been fully described in Chapter 20. A further protocol for the rapid identification of vaccine-associated rabies cases from field brain samples, targeting the viral glycoprotein (G) gene, is presented herein. Such a protocol could be used in association with the lyssavirus-typing protocol previously mentioned, and is a valid and sensitive alternative to monoclonal antibody (MAb) typing within the framework of a post-vaccination survey.
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- 2014
25. Lyssavirus detection and typing using pyrosequencing
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Hervé Bourhy, Silvia Tiozzo Caenazzo, Paola De Benedictis, Laurent Dacheux, Giovanni Cattoli, Silvia Ormelli, Ilaria Capua, Sabrina Marciano, Cristian De Battisti, Anthony Lepelletier, Angela Salomoni, National Collaborating Centre for Diseases at the Animal-Human Interface, Istituto Zooprofilattico Sperimentale delle Venezie (IZSVe), Centre Collaborateur de l'OMS pour la Rage - Dynamique des lyssavirus et adaptation à l'hôte (CC-OMS), Institut Pasteur [Paris], Centre National de Référence de la Rage-Dynamique des Lyssavirus et adaptation à l'hôte (CNR), This study was partially funded by the Italian Ministry of Health(RC 29/2007) and by the European Union through the FP6/RABMEDCONTROL program, and Institut Pasteur [Paris] (IP)
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Microbiology (medical) ,Rabies ,medicine.disease_cause ,Sensitivity and Specificity ,03 medical and health sciences ,symbols.namesake ,Viral Proteins ,Virology ,medicine ,Animals ,Humans ,Typing ,Viral ,Lyssavirus ,030304 developmental biology ,Sanger sequencing ,0303 health sciences ,biology ,030306 microbiology ,Reverse Transcriptase Polymerase Chain Reaction ,Rabies virus ,RNA virus ,DNA ,Sequence Analysis, DNA ,Rhabdoviridae ,medicine.disease ,biology.organism_classification ,3. Good health ,Nucleoproteins ,[SDV.MP.VIR]Life Sciences [q-bio]/Microbiology and Parasitology/Virology ,symbols ,Pyrosequencing ,RNA ,RNA, Viral ,Sequence Analysis - Abstract
Rabies is a fatal zoonosis caused by a nonsegmented negative-strand RNA virus, namely, rabies virus (RABV). Apart from RABV, at least 10 additional species are known as rabies-related lyssaviruses (RRVs), and some of them are responsible for occasional spillovers into humans. More lyssaviruses have also been detected recently in different bat ecosystems, thanks to the application of molecular diagnostic methods. Due to the variety of the members of the genus Lyssavirus , there is the necessity to develop a reliable molecular assay for rabies diagnosis able to detect and differentiate among the existing rabies and rabies-related viruses. In the present study, a pyrosequencing protocol targeting the 3′ terminus of the nucleoprotein (N) gene was applied for the rapid characterization of lyssaviruses. Correct identification of species was achieved for each sample tested. Results from the pyrosequencing assay were also confirmed by those obtained using the Sanger sequencing method. A pan-lyssavirus one-step reverse transcription (RT)-PCR was developed within the framework of the pyrosequencing procedure. The sensitivity (Se) of the one-step RT-PCR assay was determined by using in vitro -transcribed RNA and serial dilutions of titrated viruses. The assay demonstrated high analytical and relative specificity (Sp) (98.94%) and sensitivity (99.71%). To date, this is the first case in which pyrosequencing has been applied for lyssavirus identification using a cheaper diagnostic approach than the one for all the other protocols for rapid typing that we are acquainted with. Results from this study indicate that this procedure is suitable for lyssavirus detection in samples of both human and animal origin.
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- 2011
26. Rabies and Canine Distemper Virus Epidemics in the Red Fox Population of Northern Italy (2006-2010)
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Monica Lorenzetto, Christopher J. Rhodes, Christl A. Donnelly, Giovanni Cattoli, Manuela Dalla Pozza, Federica Obber, Marco De Nardi, Pierre Nouvellet, C. Citterio, Simon Cauchemez, Paola De Benedictis, and Medical Research Council (MRC)
- Subjects
Veterinary medicine ,Vulpes ,Epidemiology ,animal diseases ,lcsh:Medicine ,Foxes ,Population Modeling ,SUSCEPTIBILITY ,Wildlife ,medicine.disease_cause ,EMERGENCE ,0403 veterinary science ,lcsh:Science ,Distemper Virus, Canine ,0303 health sciences ,education.field_of_study ,Multidisciplinary ,biology ,Transmission (medicine) ,Zoonotic Diseases ,04 agricultural and veterinary sciences ,PREVALENCE ,3. Good health ,Vaccination ,VULPES-VULPES ,Italy ,Veterinary Diseases ,Epidemiological Monitoring ,Science & Technology - Other Topics ,VACCINATION ,WILD CARNIVORES ,Research Article ,EUROPE ,General Science & Technology ,040301 veterinary sciences ,Rabies ,Animal Types ,Population ,Infectious Disease Epidemiology ,Veterinary Epidemiology ,03 medical and health sciences ,SURVEILLANCE ,MD Multidisciplinary ,medicine ,Animals ,Distemper ,Rabies transmission ,education ,Epidemics ,Biology ,030304 developmental biology ,EMERGING INFECTIOUS-DISEASES ,Science & Technology ,Models, Statistical ,Population Biology ,MULTIDISCIPLINARY SCIENCES ,Canine distemper ,lcsh:R ,Rabies virus ,Computational Biology ,medicine.disease ,biology.organism_classification ,ANTIBODIES ,lcsh:Q ,Veterinary Science ,Infectious Disease Modeling - Abstract
Since 2006 the red fox (Vulpes vulpes) population in north-eastern Italy has experienced an epidemic of canine distemper virus (CDV). Additionally, in 2008, after a thirteen-year absence from Italy, fox rabies was re-introduced in the Udine province at the national border with Slovenia. Disease intervention strategies are being developed and implemented to control rabies in this area and minimise risk to human health. Here we present empirical data and the epidemiological picture relating to these epidemics in the period 2006-2010. Of important significance for epidemiological studies of wild animals, basic mathematical models are developed to exploit information collected from the surveillance program on dead and/or living animals in order to assess the incidence of infection. These models are also used to estimate the rate of transmission of both diseases and the rate of vaccination, while correcting for a bias in early collection of CDV samples. We found that the rate of rabies transmission was roughly twice that of CDV, with an estimated effective contact between infected and susceptible fox leading to a new infection occurring once every 3 days for rabies, and once a week for CDV. We also inferred that during the early stage of the CDV epidemic, a bias in the monitoring protocol resulted in a positive sample being almost 10 times more likely to be collected than a negative sample. We estimated the rate of intake of oral vaccine at 0.006 per day, allowing us to estimate that roughly 68% of the foxes would be immunised. This was confirmed by field observations. Finally we discuss the implications for the eco-epidemiological dynamics of both epidemics in relation to control measures.
- Published
- 2013
- Full Text
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27. Rabies in a vaccinated dog in Italy
- Author
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C. Veggiato, Gaetana Ferri, Franco Mutinelli, Paola De Benedictis, Glauco Squecco, Ilaria Capua, and Renato Coassin
- Subjects
medicine.medical_specialty ,Veterinary medicine ,General Veterinary ,business.industry ,Dachshund ,Public health ,Prevalence ,MEDLINE ,General Medicine ,medicine.disease ,Northern italy ,Vaccination ,Family medicine ,Epidemiology ,medicine ,Rabies ,business - Abstract
A seven-year-old dachshund died suddenly at a veterinary clinic in northern Italy on June 18, 2009, after showing unnatural aggressive behaviour. The epidemiological investigation reported that the dog had not bitten people or animals in the previous month, or shown clinical signs apart from the
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