ORIGINAL PAPER Is Acarapis woodi a single species? A new PCR protocol to evaluate its prevalence A. Cepero & R. Martín-Hernández & L. Prieto & T. Gómez-Moracho & A. Martínez-Salvador & C. Bartolomé & X. Maside & A. Meana & M. Higes Received: 29 October 2014 /Accepted: 6 November 2014 /Published online: 18 November 2014 # Springer-Verlag Berlin Heidelberg 2014 Abstract 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 se- quences 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. Keywords Parasites . Acarapis woodi . Diagnosis . Genetic variability . Honey bee colony losses Introduction Acarapisosis is a disease of the adult honey bee (Apis mellifera) caused by the microscopic tracheal mite, Acarapis woodi (Rennie), which primarily lives and reproduces in the large prothoracic trachea (OIE 2008). The pathogenic effect of this mite on individual bees depends on the number of para- sites within the tracheae. Indeed, they can be attributed to both mechanical injuries and to physiological disorders resulting from the obstruction of the air ducts, to lesions in the tracheal A. Cepero : R. Martín-Hernández : T. Gómez-Moracho : M. Higes (*) Laboratorio de Patología Apícola, Consejería de Agricultura, Junta de Comunidades de Castilla-La Mancha, Centro Apícola Regional (CAR), San Martín s/n. 19180 Marchamalo, Guadalajara, Spain e-mail: mhiges@jccm.es R. Martín-Hernández Fundación Parque Científico y Tecnológico de Albacete (INCRECYT- FEDER), Madrid, Spain A. Martínez-Salvador C/ Puente la Reina, 28050 Madrid, Spain C. Bartolomé : X. Maside Departamento de Anatomía Patolóxica e Ciencias Forenses, Universidade de Santiago de Compostela, 15782, Santiago de Compostela, Galicia, Spain C. Bartolomé : X. Maside Grupo de Medicina Xenómica, CIMUS, Universidade de Santiago de Compostela, 15782 Santiago de Compostela, Galicia, Spain C. Bartolomé : X. Maside Grupo de Xenómica Comparada de Parásitos Humanos, IDIS, Santiago de Compostela, Galicia, Spain A. Meana Animal Health Department, Facultad de Veterinaria, Universidad Complutense, 28040 Madrid, Spain L. Prieto Comisaría General de la Policía Científica, DNA Laboratory, Instituto Universitario de Investigación en ciencias policiales (IUICP), Madrid, Spain Parasitol Res (2015) 114:651–658 DOI 10.1007/s00436-014-4229-6