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Martin-Hernandez R.,Centro Apicola Regional | Martin-Hernandez R.,Institute Recursos Humanos para la Ciencia y la Tecnologia INCRECYT | Botias C.,Centro Apicola Regional | Bailon E.G.,Centro Apicola Regional | And 4 more authors.
Environmental Microbiology | Year: 2012

Nosema ceranae has been suggested to be replacing Nosema apis in some populations of Apis mellifera honeybees. However, this replacement from one to the other is not supported when studying the distribution and prevalence of both microsporidia in professional apiaries in Spanish territories (transverse study), their seasonal pattern in experimental hives with co-infection or their prevalence at individual level (either in worker bees or drones). Nevertheless, N. ceranae has shown to present a higher prevalence at all the studied levels that could indicate any advantage for its development over N. apis or that it is more adapted to Spanish conditions. Also, both microsporidia show a different pattern of preference for its development according to the prevalence in the different Spanish bioclimatic belts studied. Finally, the fact that all analyses were carried out using an Internal PCR Control (IPC) newly developed guarantees the confidence of the data extracted from the PCR analyses. This IPC provides a useful tool for laboratory detection of honeybee pathogens. © 2011 Society for Applied Microbiology and Blackwell Publishing Ltd. Source


Cepero A.,Centro Apicola Regional CAR | Martin-Hernandez R.,Centro Apicola Regional CAR | Prieto L.,Instituto Universitario Of Investigacion En Ciencias Policiales Iuicp | Gomez-Moracho T.,Centro Apicola Regional CAR | And 4 more authors.
Parasitology Research | Year: 2014

Acarapisosis is a disease of the adult honey bee Apis mellifera L., caused by the tracheal mite Acarapis woodi (Rennie), that affects the prothoracic tracheas of worker honey bees. Although it is not usually considered a real problem for honey bee colonies in southern Europe (mainly Spain and Greece), where the majority of professional beekeepers are located in Europe, recent works have reported the constant presence of this mite in this area, making it a potential cofactor for colony losses. In this study, we developed a specific PCR diagnostic tool that improves the techniques used so far and allowed us to confirm the presence of this parasite in Spain, urging the need to monitor its prevalence and implications in the health of the colonies. Indeed, in a total of 635 apiaries analysed, the prevalence of A. woodi in 2010 was 8.3 and 4 % in 2011. The mite is present in bee colonies over time and should not be underestimated as a possible cofactor in the collapse of bee colonies. Additionally, some positive samples were cloned so a genetic analysis on the diversity within A. woodi isolates was also approached. This allowed us to identify different genetic variants within an isolate, even when they were present at low frequencies. And this genetic analysis revealed the existence of a different clade of Acarapis sequences that could represent a new species or subspecies, although more research is required to verify the identity of this novel lineage at genetic and morphological level. © 2014, Springer-Verlag Berlin Heidelberg. Source


Garrido-Bailon E.,Centro Apicola Regional CAR | Higes M.,Centro Apicola Regional CAR | Martinez-Salvador A.,Epidemiology Consultant | Antunez K.,Institute Investigaciones Biologicas Clemente Estable | And 5 more authors.
Microbial Biotechnology | Year: 2013

The microorganisms Ascosphaera apis, Paenibacillus larvae and Melissococcus plutonius are the three most important pathogens that affect honeybee brood. The aim of the present study was to evaluate the prevalence of these pathogens in honeybee colonies and to elucidate their role in the honeybee colony losses in Spain. In order to get it, a multiplex polymerase chain reaction (PCR) assay was developed to simultaneously amplify the16S ribosomal ribonucleic acid (rRNA) gene of P. larvae and M. plutonius, and the 5.8S rRNA gene of A. apis. The multiplex PCR assay provides a quick and specific tool that successfully detected the three infectious pathogens (P. larvae, M. plutonius and A. apis) in brood and adult honeybee samples without the need for microbiological culture. This technique was then used to evaluate the prevalence of these pathogens in Spanish honeybee colonies in 2006 and 2007, revealing our results a low prevalence of these pathogens in most of the geographic areas studied. © 2013 The Authors. Microbial Biotechnology published by John Wiley & Sons Ltd and Society for Applied Microbiology. Source


Montesino M.,Instituto Universitario Of Investigacion En Ciencias Policiales Iuicp | Prieto L.,Instituto Universitario Of Investigacion En Ciencias Policiales Iuicp
Methods in Molecular Biology | Year: 2012

Cycle sequencing reaction with Big-Dye terminators provides the methodology to analyze mtDNA Control Region amplicons by means of capillary electrophoresis. DNA sequencing with ddNTPs or terminators was developed by (1). The progressive automation of the method by combining the use of fluorescent-dye terminators with cycle sequencing has made it possible to increase the sensibility and efficiency of the method and hence has allowed its introduction into the forensic field. PCR-generated mitochondrial DNA products are the templates for sequencing reactions. Different set of primers can be used to generate amplicons with different sizes according to the quality and quantity of the DNA extract providing sequence data for different ranges inside the Control Region. © 2012 Springer Science+Business Media, LLC. Source

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