Investigation of an Autochthonous Outbreak of Bovine Besnoitiosis in Northwestern Sicily

Abstract Bovine besnoitiosis is a parasitic disease caused by the protozoan Besnoitia besnoiti, leading to infertility in bulls and abortions in cows. In Italy, it is considered an emerging disease, recently introduced by the importation of animals from Spain and France. In the last decade, many outbreaks have been reported and confirmed in native cattle, mostly in northern and central Italy. This study reports on an autochthonous outbreak of bovine besnoitiosis in two nearby farms located in northwestern Sicily. A 15-month-old Limousine bull born on the farm showed typical clinical signs of the chronic disease phase, such as edema of the scrotum with subsequent hyperkeratosis associated with eschars and thickening of the skin. A histopathological examination revealed the presence of Besnoitia tissue cysts containing bradyzoites in the eyes, tendons, testicles, dermis, and nictitating membrane. A serological investigation using a commercial ELISA kit revealed a high seroprevalence of the antibody anti-B. besnoiti (79.2% for the farms in this study). Clinical disease showed low prevalence (1.5%) despite the high seroprevalence of specific antibodies in the herd, confirming that bovine besnoitiosis is an emergent endemic pathogen in Sicily, but its clinical behavior still remains sporadic

BOVINE BESNOITIOSIS: A CASE REPORT IN SICILY

IINTRODUCTION Bovine besnoitiosis is a parasitic disease caused by the tissue cyst forming protozoan Besnoitia besnoiti The European Food Safety Authority ( has reported that bovine besnoitiosis is re emerging in Europe 1 It represents a chronic and debilitating disease that is responsible for severe economic losses such as poor body condition, decreased milk production, infertility in bulls and occasional abortion 2 Autochthonous outbreaks of the disease were reported in Central Italy for the first time in 2009 3 In Sicily, although the disease was observed in a heifer in 2014 4 no epidemiological data is known AIM OF THE WORK This study describes an autochthonous outbreak of Bovine besnoitiosis in North Western Sicily a bull that shows typical clinical symptoms related to bovine besnoitiosis and moreover, the serological investigation of the herd, suspected of being infected by B besnoiti MATERIALS AND METHODS In a 16 month old affected bull, clinical status was evaluated both in acute and chronic stage After slaughtering, samples of skin, conjunctiva, tongue, and tendons were collected for histopathological examination Furthermore, i n the bull and in the Limousine herd of 72 animals, serological investigation using ID Screen® Besnoitia Indirect 2 0 ID vet was performed. RESULT The bull showed typical clinical signs of besnoitiosis such as: high fever, diffuse oedema at the joints of limbs and scrotum, hyperemia of muzzle and eyes, orchitis , thickening of the nasal planum and wheezing. About 1 0 12 days later, skin lesions ( hyperkeratosis, alopecia, dandruff and scab) appeared (Figure 1). At the 58th day after the beginning of the first signs, the animal showed diffuse skin thickening, was emaciated and reluctant to move. Histological examination showed multifocal coalescing thick wall cysts surrounding parasitophorous vacuole containing thousands of banana shaped bradyzoites all the collected samples (Figure 2 3). ELISA confirmed the seropositivity of the bull, moreover, 79,17% of the herd resulted positive (Tab 1) CONCLUSION. Finally, since all the animals were born in the farm or were bought from nearby farms this case confirms the presence of autochthonous B. besnoiti also in Sicily and may contribute to a further epidemiological study in this area. REFERENCES [1] EFSA Journal, 2010, EFSA Journal 8: 1499-514 [2]Gutiérrez Expósito et al., 2017 Int J Parasitol . 47: 737 51 [3]A. Gentile et al., 2012 Vet Parasitol . 184: 108 15 [4]Di Marco et al., 2016 LXX SISVET. 430

Detección y caracterización molecular del SARS-CoV- 2 en animales

The zoonotic origin of the SARS-CoV-2 has raised the necessity to monitor and control the virus in domestic and wild animals to determine the possible role of the animals as carriers, reservoirs and / or virus amplifiers. Studies to understand the susceptibility of different animals to SARS-CoV-2 and to evaluate the dynamics of the infection, identification of the possible reservoirs and / or transmitters, are fundamental tools for the surveillance of the pandemic. The design of specific protocols for the epidemiological survey and for taking samples of pets belonging to positive COVID-19 owners began the work. The goals of the project involve the diagnosis and epidemiological control of SARS-CoV-2 in animals that live with infected people, the analysis of the potential infectivity of the virus and the role that animals play in the pandemic. These studies are an instrument that will allow us to know the infectivity of the virus in different hosts and its implications in the transmission and perpetuation of the disease. The project began with the design of protocols for animal sampling, defining a suspected case and specifying how a particular veterinarian should proceed when asked by the positive SARS-CoV-2 owners. Said protocol was published by the Veterinary College of the Buenos Aires Province (District II). An informed agreement was created, approved by the Institutional Commission for the Care and Use of Laboratory Animals (CICUAL). The agreement is signed by those responsible for the animals and the project director, approving to take samples. On the other hand, the zoonosis area dependent on the National Ministry of Health adopted the protocols and is working on the implementation of an epidemiological surveillance system. The project contents and objectives was distributed in various media, allowing those SARS-CoV-2 owners diagnosed positive to communicate with members of the project for consultations and coordination for pets sampling. To August 31, rectal and oropharyngeal swabs were collected from 15 canines and 11 felines, all in close contact with people diagnosed positive for SARS-CoV-2. In addition, samples were collected from a chimpanzee from Ecopaque La Plata. Once in the laboratory, the RNA was extracted from the samples using a commercial kit, and the molecular diagnosis by real-time RT-PCR was carried out adapting commercial kits. The samples tested were non-reactive to the specific targets of SARS-CoV-2.El origen zoonótico del SARS-CoV-2 ha planteado la necesidad de realizar el monitoreo y control de esta virosis en animales domésticos y silvestres para determinar el posible rol de los animales como portadores, reservorios y/o amplificadores del virus. Los estudios para comprender mejor la susceptibilidad de diferentes especies animales al SARS-CoV-2 y evaluar la dinámica de la infección en estas especies, así como también la identificación de posibles reservorios y/o transmisores, son una herramienta fundamental para la vigilancia de la pandemia.  El diseño de protocolos específicos para el relevamiento epidemiológico y la toma de muestras de animales pertenecientes a pacientes positivos a COVID-19, dieron inicio al trabajo. Los objetivos del proyecto involucran el diagnóstico y la vigilancia epidemiológica del SARS-CoV-2 en animales que conviven con personas infectadas, para analizar la infectividad potencial del virus y el rol que cumplen los animales en la pandemia. Estos estudios son una herramienta que nos permitirá conocer el comportamiento del virus en diferentes hospedadores y sus implicancias en la transmisión y perpetuación de la enfermedad. El proyecto comenzó con el diseño de protocolos para toma de muestras, definiendo caso sospechoso en animales, e indicando cómo debe proceder el Médico Veterinario particular al ser consultado por los propietarios. Dicho protocolo fue difundido por el Colegio de Veterinarios de la Provincia de Buenos Aires (Distrito II). Conjuntamente, se creó un consentimiento informado, aprobado por la Comisión Institucional para el Cuidado y Uso de Animales de Laboratorio (CICUAL) que es firmado por los responsables de los animales y del proyecto, acordando la toma de muestras. Por otro lado, el área de zoonosis dependiente del Ministerio de Salud de la Nación adoptó los protocolos y trabaja en la implementación de un sistema de vigilancia epidemiológica. Se realizó la difusión del proyecto en diversos medios de comunicación, permitiendo que aquellos pacientes diagnosticados positivos a SARS-CoV-2 se comunicaran con el grupo de trabajo para consultas y coordinación de la toma de muestras a sus mascotas. Hasta el 31 de agosto, se tomaron muestras de hisopados orofaríngeos y rectales de 15 caninos y 11 felinos, todos en estrecho contacto con personas diagnosticados positivos a SARS-CoV-2. Además, se colectaron muestras de un chimpancé del Ecoparque La Plata. Una vez en el laboratorio se realizó la extracción del ARN de las muestras utilizando un kit comercial, y el diagnóstico molecular por real-time RT-PCR adaptando kits comerciales. Las muestras analizadas fueron no reactivas a los targets específicos de SARS-CoV-2

Detection and molecular characterization of SARS-CoV-2 in animals

El potencial origen zoonótico del SARS-CoV-2 ha planteado la necesidad de realizar el monitoreo y control de esta virosis en animales domésticos y silvestres para determinar el posible rol de los animales como portadores, reservorios y/o amplificadores del virus. Los estudios para comprender mejor la susceptibilidad de diferentes especies animales al SARS-CoV-2 y evaluar la dinámica de la infección en estas especies, así como también la identificación de posibles reservorios y/o transmisores, son una herramienta fundamental para la vigilancia de la pandemia. El diseño de protocolos específicos para el relevamiento epidemiológico y la toma de muestras de animales pertenecientes a pacientes positivos a COVID-19, dieron inicio al trabajo. Los objetivos del proyecto involucran el diagnóstico y la vigilancia epidemiológica del SARS-CoV-2 en animales que conviven con personas infectadas, para analizar la infectividad potencial del virus y el rol que cumplen los animales en la pandemia. Estos estudios son una herramienta que nos permitirá conocer el comportamiento del virus en diferentes hospedadores y sus implicancias en la transmisión y perpetuación de la infección.The zoonotic potential origin of the SARS-CoV-2 has raised the necessity to monitor and control the virus in domestic and wild animals to determine the possible role of the animals as carriers, reservoirs and / or virus amplifiers. Studies to understand the susceptibility of different animals to SARS-CoV-2 and to evaluate the dynamics of the infection, identification of the possible reservoirs and / or transmitters, are fundamental tools for the surveillance of the pandemic. The design of specific protocols for the epidemiological survey and for taking samples of pets belonging to positive COVID-19 owners began the work. The goals of the project involve the diagnosis and epidemiological control of SARS-CoV-2 in animals that live with infected people, the analysis of the potential infectivity of the virus and the role that animals play in the pandemic. These studies are an instrument that will allow us to know the infectivity of the virus in different hosts and its implications in the transmission and perpetuation of the infection.El proyecto es financiado por la Agencia Nacional de Promoción de la Investigación, el Desarrollo Tecnológico y la Innovación, por la Convocatoria Extraordinaria a Ideas Proyecto IP-COVID-19. IP COVID 19 - 468 - Detección y caracterización molecular del SARS-CoV-2 en animales.Secretaría de Ciencia y Técnic

Primer reporte de SARS-CoV-2 detectado en un gato de Argentina

El síndrome respiratorio agudo severo coronavirus 2 (SARS-CoV-2) es el responsable de la pandemia de COVID-19 (coronavirus disease 2019). Los diferentes análisis realizados hacen suponer que se originó a partir de coronavirus de murciélago; sin embargo, aún se desconoce el animal que podría haber actuado como intermediario. Los coronavirus son una preocupación en salud pública por su potencial zoonótico, capaz de causar nuevos brotes de enfermedades. La diversidad genética, debida a la alta frecuencia de mutación y los eventos de recombinación, probablemente esté relacionada con la variedad de hospedadores. Este hecho plantea la necesidad de realizar el monitoreo de animales para determinar el posible rol que cumplen como hospedadores naturales, vectores o reservorios del virus, permitiendo evaluar la dinámica de la infección y considerándose una herramienta fundamental para la vigilancia epidemiológica. Hasta el momento, se han reportado detecciones de infección en felinos, caninos, hurones y visones, lo que demuestra que varias especies de animales son susceptibles al SARS-CoV-2. Asimismo, se reportó la presencia de anticuerpos específicos en sueros de caninos y felinos. El objetivo de este trabajo fue detectar la presencia de SARS-CoV-2 en mascotas que habían estado en contacto estrecho con sus dueños, previamente confirmados como COVID-19 positivos.Trabajo publicado en Cagliada, Maria del Pilar Lilia y Galosi, Cecilia Mónica (comps.). I Congreso de Microbiología Veterinaria. Libro de resúmenes. La Plata: Facultad de Ciencias Veterinarias, 2021.Facultad de Ciencias Veterinaria

Safety of procuring research tissue during a clinically indicated kidney biopsy from patients with lupus: data from the Accelerating Medicines Partnership RA/SLE Network

Objectives In lupus nephritis the pathological diagnosis from tissue retrieved during kidney biopsy drives treatment and management. Despite recent approval of new drugs, complete remission rates remain well under aspirational levels, necessitating identification of new therapeutic targets by greater dissection of the pathways to tissue inflammation and injury. This study assessed the safety of kidney biopsies in patients with SLE enrolled in the Accelerating Medicines Partnership, a consortium formed to molecularly deconstruct nephritis.Methods 475 patients with SLE across 15 clinical sites in the USA consented to obtain tissue for research purposes during a clinically indicated kidney biopsy. Adverse events (AEs) were documented for 30 days following the procedure and were determined to be related or unrelated by all site investigators. Serious AEs were defined according to the National Institutes of Health reporting guidelines.Results 34 patients (7.2%) experienced a procedure-related AE: 30 with haematoma, 2 with jets, 1 with pain and 1 with an arteriovenous fistula. Eighteen (3.8%) experienced a serious AE requiring hospitalisation; four patients (0.8%) required a blood transfusion related to the kidney biopsy. At one site where the number of cores retrieved during the biopsy was recorded, the mean was 3.4 for those who experienced a related AE (n=9) and 3.07 for those who did not experience any AE (n=140). All related AEs resolved.Conclusions Procurement of research tissue should be considered feasible, accompanied by a complication risk likely no greater than that incurred for standard clinical purposes. In the quest for targeted treatments personalised based on molecular findings, enhanced diagnostics beyond histology will likely be required

Seroprevalence of Coxiella burnetii in cattle and in operators at risk of infection in Sicily

The aim of this survey was to access: the presence of abortigens agents in a dairy cattle farm and the Coxiella burnetii seroprevalence in operators at risk in Sicil

Indagine sulla prevalenza periodale della BVD-MD e incidenza di questa sul benessere della mandria

At quarterly intervals, 5 blood samples were carried out in all bovine animals present in a cattle farm of about 100 animals. This farm had already had some positivity to the Pestivirus that causes BVD-MD. This was to check if there was a correlation between climate change and viral circulation

Prevalence and Genotypes of Anaplasma Species and Habitat Suitability for Ticks in a Mediterranean Ecosystem▿

Anaplasma species are tick-transmitted pathogens that impact veterinary and human health. Sicily is one of the locations where these pathogens are endemic. Sicily represents a typical Mediterranean ecosystem to study Anaplasma infection and tick habitat suitability. The aims of this study were (i) to characterize by 16S rRNA and species-specific msp4 gene PCR the prevalence and genotypes of A. marginale, A. phagocytophilum, and A. ovis in the most abundant host species in Sicilian provinces and (ii) to correlate differences between hosts and between western and eastern Sicily with the habitat suitability for ticks in these regions. Differences were found in the prevalence of Anaplasma spp. between different hosts and between western and eastern provinces. The differences in Anaplasma prevalence between different hosts may be explained by pathogen host tropism. The differences between western and eastern provinces correlated with the tick habitat suitability in these regions. The analysis of Anaplasma genotypes suggested a higher host and regional specificity for A. phagocytophilum than for A. marginale and A. ovis strains, a finding probably associated with the broader host range of A. phagocytophilum. The presence of identical A. marginale genotypes in the two regions may reflect cattle movement. The results for A. ovis suggested the possibility of some genotypes being host specific. These results provide information potentially useful for the management of tick-borne diseases caused by Anaplasma spp. in Sicily and other Mediterranean regions and may contribute to the development of models to predict the risks for these tick-borne pathogens

Prevalence of Anaplasma species and habitat suitability for ticks in Sicily

The genus Anaplasma (Rickettsiales: Anaplasmataceae) includes the tick-transmitted pathogens Anaplasma marginale, Anaplasma phagocytophilum and Anaplasma ovis that have an impact on veterinary and human health. Anaplasma spp. are endemic in Sicily, where they constitute a veterinary and human health problem. Sicily represents a typical Mediterranean ecosystem and offers a good scenario to study the prevalence of Anaplasma spp. in relation to the habitat suitability for ticks. The aim of this study was to characterise the prevalence of Anaplasma spp. in most abundant host species in Sicily and to correlate these data with the habitat suitability (HS) for ticks at the province level.This work was supported by project IZS SI 05 ⁄ 04 of the Ministry of Health, Italy. Partial support was also provided by the Ministerio de Ciencia e Innovacion, Spain (project BFU2008-01244 ⁄ BMC).Peer reviewe