Simulaciones sanitarias para el estudio de enfermedades infecciosas endémicas, emergentes y reemergentes en el ganado porcino

Depto. de Sanidad AnimalFac. de VeterinariaFALSEsubmitte

Aplicación de la termografía en la valoración de la fertilidad en huevos de Cernícalo primilla

Habitualmente la valoración de la fertilidad de los huevos se realiza mediante un ovoscopio. El principal inconveniente es la necesidad de manipulación del huevo, que impide su aplicación a la fauna salvaje. Por ello se estudió su posible sustitución por termografía, analizándose los patrones térmicos de 190 huevos de Cernícalo primilla, Falco naumanni. Los resultados obtenidos muestran que la termografía nos permite diferenciar los huevos en fértiles, infértiles y abortados gracias a sus diferentes patrones térmicos y velocidades de enfriamiento desde tres días después de la puesta. Es posible emplearlo tanto durante incubación artificial como en la naturaleza con diferentes condiciones ambientales, siendo necesario tener en cuenta el lugar de medición, los días transcurridos desde la puesta así como la temperatura de partida.Usually the evaluation of eggs fertility is done with ovoscopy. The principal inconvenient is the need to manipulate the egg, this prevents its application in wildlife. This is why it was studied to substitute this method for termography, analizing the termic patterns of 190 eggs of Falco naumanni. The obtained results show that termography allows us to differentiate the fertile eggs, from unfertile, and aborted ones thanks to its different termic patters and cooling speeds since 3 days after the lay. Its possible to use it both during the artificial incubation and in nature with different environment conditions, being necessary to have in mind the place, the number of days that have passed since the lay as well as the temperature at the beginning

El papel de la fauna silvestre en las enfermedades emergentes

Las enfermedades infecciosas zoonósicas emergentes se han convertido en la actualidad en una de las amenazas más graves para la salud pública. Aproximadamente el 75 % de las enfermedades que han surgido durante las últimas dos décadas tienen su fuente en la fauna salvaje. Las recientes epidemias del síndrome agudo respiratorio severo (SARS), infecciones por virus West Nile o influenza aviar, que serán tratadas en este trabajo, demuestran la importancia de las enfermedades emergentes en todo el mundo y el relevante papel de los servicios veterinarios. Para vigilar estas enfermedades es preciso disponer de un equipo de trabajo multidisciplinar, con profesionales formados en medicina, epidemiología, virología, zoología y veterinaria, por la gran interrelación de factores. Asimismo, siempre hay que tener en cuenta en estas enfermedades tan contagiosas todas las vías de transmisión, no sólo animales, sino los movimientos comerciales tanto legales como ilegales.Zoonotic emergent infectious diseases have become one of the most serious threats for public health. Approximately 75 % of the diseases that have occurred during the last two decades originated in the wild fauna. SARS, West Nile virus or influenza recent epidemics, that will be treated in this article, show the importance of emerging diseases in the whole world and the relevant role of the veterinary services. Monitoring these diseases requires a multidisciplinary work team, with professionals formed in medicine, epidemiology, virology, zoology and veterinary sciences, because of the great interrelationship of factors. Likewise, it is always important to bear all the routes of transmission in mind in these very contagious diseases, not only animals, but the commercial, both legal and illegal, movements

Estudio molecular de una nueva cepa de morbillivirus de cetáceo aislada de un calderon tropical

El morbillivirus de cetáceo (CeMV) es el virus más patógeno de cetáceos. Comprende tres cepas cuyos nombres provienen de la especie de donde se aisló por primera vez: el morbillivirus de delfín (DMV), el morbillivirus de la marsopa (PMV) y el morbillivirus del calderón (PWMV). En este estudio se pretende completar la caracterización molecular de una nueva cepa de CeMV obtenida del encéfalo de un calderón tropical que varó muerto en las Islas Canarias, con lesiones compatibles con enfermedad por morbillivirus. La construcción de árboles filogenéticos con todas las especies de morbillivirus indicó una mayor homología con PWMV por lo que se propone llamar esta nueva cepa “PWMV II”. Además, en base a los árboles filogenéticos y una exhaustiva revisión bibliográfica, se hace la propuesta de renombrar las tres cepas de CeMV en “CeMV-1” para el DMV, “CeMV-2” para el PMV y “CeMV-3” para el PWMV.Cetacean morbillivirus (CeMV) is the most pathogenic virus in cetaceans. It includes three strains whose names refer to the species of odontocete from which it was initially isolated: Dolphin morbillivirus (DMV), Porpoise morbillivirus (PMV) and Pilot Whale morbillivirus (PWMV). The aim of this work was to complete a molecular characterization of a new strain of CeMV obtained from the brain of a short finned pilot whale stranded dead around Canary Islands with lesions compatible with morbilliviral disease. Phylogenetic trees including sequences of all the morbillivirus species have shown a higher homology with PWMV, so this strain is tentatively named “PWMV II”. Moreover, from the phylogenetic trees analysis and an exhaustive bibliographical review we propose new names for the CeMV strains:“CeMV-1” for DMV, “CeMV-2” for PMV and “CeMV-3” for PWMV

Detecciónes de la expresión de citoquinas mediante RT-PCR en tiempo real en ganado ovino

Las citoquinas son moléculas proteicas secretadas por diferentes células, fundamentalmente del sistema inmunitario, como respuesta a una estimulación inmunológica. Existen diferentes técnicas para estudiar la producción y acción de éstas en los individuos. Este estudio consiste en la puesta a punto de una técnica de Retrotranscripción- Reacción en cadena de la polimerasa (RT-PCR) en tiempo real para la detección de la expresión del ARNm que codifica seis citoquinas (TNF-α, IFNγ, IL-4, IL2, IL-10, IL-12). Los resultados preliminares se obtuvieron tras la aplicación de la técnica a linfocitos estimulados in vitro con ConA (Concanavalina A) y LPS (Lipopolisacárido de E.coli) demostrando la detección de ARNm para IL-4, IL-2 e IFNγ y TNF-α, IL-12 e IL-10, respectivamente; posteriormente se realizó el estudio en ovejas inmunizadas frente a Chlamydophyla abortus, permitiendo la detección de la expresión de ARNm de las seis citoquinas. Estos resultados demuestran que es una técnica rápida, sensible y fiable para la detección de citoquinas.Cytokines are protein molecules that cells from the immune system secrete in response to immune stimulation. There are different techniques to study their production and their action on individuals. This study consists of the development of a real time RT-PCR method for the detection of the expression of mRNA coding for six cytokines (TNF-α, IFNγ, IL-4, IL2, IL-10, IL-12). The preliminary results were obtained from the study of lymphocites stimulated with Con A and LPS, showing the detection of ARNm, IL-4, IL-2, IFNγ and TNF-α, IL-12, IL-10, respectively. Its subsequent application in sheep that were vaccinated against Chlamydophila abortus, allowed the detection of mRNA expression for the six cytokines. These results show that it is a swift, sensitive and reliable technique to detect mRNA of cytokines

Characterization of the Immune Response Induced by a Commercially Available Inactivated Bluetongue Virus Serotype 1 Vaccine in Sheep

The protective immune response generated by a commercial monovalent inactivated vaccine against bluetongue virus serotype 1 (BTV1) was studied. Five sheep were vaccinated, boost-vaccinated, and then challenged against BTV1 ALG/2006. RT-PCR did not detect viremia at any time during the experiment. Except a temperature increase observed after the initial and boost vaccinations, no clinical signs or lesions were observed. A specific and protective antibody response checked by ELISA was induced after vaccination and boost vaccination. This specific antibody response was associated with a significant increase in B lymphocytes confirmed by flow cytometry, while significant increases were not observed in T lymphocyte subpopulations (CD4+, CD8+, and WC1+), CD25+ regulatory cells, or CD14+ monocytes. After challenge with BTV1, the antibody response was much higher than during the boost vaccination period, and it was associated with a significant increase in B lymphocytes, CD14+ monocytes, CD25+ regulatory cells, and CD8+ cytotoxic T lymphocytes

African swine fever virus transmission cycles in Central Europe: evaluation of wild boar-soft tick contacts through detection of antibodies against Ornithodoros erraticus saliva antigen.

BACKGROUND African swine fever (ASF) is one of the most complex viral diseases affecting both domestic and wild pigs. It is caused by ASF virus (ASFV), the only DNA virus which can be efficiently transmitted by an arthropod vector, soft ticks of the genus Ornithodoros. These ticks can be part of ASFV-transmission cycles, and in Europe, O. erraticus was shown to be responsible for long-term maintenance of ASFV in Spain and Portugal. In 2014, the disease has been reintroduced into the European Union, affecting domestic pigs and, importantly, also the Eurasian wild boar population. In a first attempt to assess the risk of a tick-wild boar transmission cycle in Central Europe that would further complicate eradication of the disease, over 700 pre-existing serum samples from wild boar hunted in four representative German Federal States were investigated for the presence of antibodies directed against salivary antigen of Ornithodoros erraticus ticks using an indirect ELISA format. RESULTS Out of these samples, 16 reacted with moderate to high optical densities that could be indicative of tick bites in sampled wild boar. However, these samples did not show a spatial clustering (they were collected from distant geographical regions) and were of bad quality (hemolysis/impurities). Furthermore, all positive samples came from areas with suboptimal climate for soft ticks. For this reason, false positive reactions are likely. CONCLUSION In conclusion, the study did not provide stringent evidence for soft tick-wild boar contact in the investigated German Federal States and thus, a relevant involvement in the epidemiology of ASF in German wild boar is unlikely. This fact would facilitate the eradication of ASF in the area, although other complex relations (wild boar biology and interactions with domestic pigs) need to be considered

African swine fever vaccine: Turning a dream into reality

African swine fever (ASF) is currently threatening the swine industry at a global level. The disease originated in Africa has spread to Europe, Asia and Oceania, since 2007, reaching a pandemic dimension. Currently, the spread of ASF is unstoppable and that the development of a safe and effective vaccine is urgently required. The objective of this paper is to review the vaccine candidates tested during the 20th and 21st centuries, to identify the strengths and weaknesses of these studies and to highlight what we should learn. Several strategies have been explored to date, some of which have shown positive and negative results. Inactivated preparations and subunit vaccines are not a viable option. The most promising strategy would appear to be live attenuated vaccines, because these vaccine candidates are able to induce variable percentages of protection against certain homologous and heterologous virus isolates. The number of studies on live attenuated vaccine candidates has steadily increased in the 21st century thanks to advances in molecular biology and an in-depth knowledge of ASF virus, which have allowed the development of vaccines based on deletion mutants. The deletion of virulence-related genes has proved to be a useful tool for attenuation, although attenuation does not always mean protection and even less, cross protection. Therefore, ASF vaccine development has proved to be one of the top priorities in ASF research. Efforts are still being made to fill the gaps in the knowledge regarding immune response, safety and cross protection, and these efforts will hopefully help to find a safe and effective vaccine that could be commercialised soon, thus making it possible to turn a dream into reality

First molecular determination of herpesvirus from two mysticete species stranded in the Mediterranean Sea

BACKGROUND Herpesvirus can infect a wide range of animal species: mammals, birds, reptiles, fish, amphibians and bivalves. In marine mammals, several alpha- and gammaherpesvirus have been identified in some cetaceans and pinnipeds species. To date, however, this virus has not been detected in any member of the Balaenoptera genus. CASE PRESENTATION Herpesvirus was determined by molecular methods in tissue samples from a male fin whale juvenile (Balaenoptera physalus) and a female common minke whale calf (Balaenoptera acutorostrata) stranded on the Mediterranean coast of the Region of Valencia (Spain). Samples of skin and penile mucosa from the fin whale and samples of skin, muscle and central nervous system tissue from the common minke whale tested positive for herpesvirus based on sequences of the DNA polymerase gene. Sequences from fin whale were identical and belonged to the Alphaherpesvirinae subfamily. Only members of the Gammaherpesvirinae subfamily were amplified from the common minke whale, and sequences from the muscle and central nervous system were identical. Sequences in GenBank most closely related to these novel sequences were viruses isolated from other cetacean species, consistent with previous observations that herpesviruses show similar phylogenetic branching as their hosts. CONCLUSIONS To our knowledge, this is the first molecular determination of herpesvirus in the Balaenoptera genus. It shows that herpesvirus should be included in virological evaluation of these animals