Reliability of Morphological and PCR Methods for the Official Diagnosis of Aethina tumida (Coleoptera: Nitidulidae): A European Inter-Laboratory Comparison

The Small Hive Beetle (Aethina tumida Murray, 1867) is an invasive scavenger of honeybees. Originally endemic in sub-Saharan Africa, it is regulated internationally in order to preserve the areas still free from this species. To ensure the reliability of official diagnoses in case of introduction, an inter-laboratory comparison was organised on the identification of A. tumida by morphology and real-time PCR. Twenty-two National Reference Laboratories in Europe participated in the study and analysed 12 samples with adult coleopterans and insect larvae. The performance of the laboratories was evaluated in terms of sensitivity and specificity. Sensitivity was satisfactory for all the participants and both types of methods, thus fully meeting the diagnostic challenge of confirming all truly positive cases as positive. Two participants encountered specificity problems. For one, the anomaly was minor whereas, for the other, the issues concerned a larger number of results, especially real-time PCR, which probably were related to inexperience with this technique. The comparison demonstrated the reliability of official diagnosis, including the entire analytical process of A. tumida identification: from the first step of the analysis to the expression of opinions. The performed diagnostic tools, in parallel with field surveillance, are essential to managing A. tumida introduction.

Updates on the Mobile Divider and Its Use in Calabria Region to Monitor and Control Aethina tumida Infestation

The small hive beetle (SHB), Aethina tumida was first detected in the Calabria and Sicily region (Southern Italy) in 2014. In this regard, a more effective and faster inspection method than the Official one (Ministry of Health) was tested to detect the beetle in the hive. In collaboration with Calabria beekeepers’ association, a “mobile divider”-based method was tested, in order to facilitate the detection of A. tumida and save time during hive inspection. In this short communication, we provide an update on the mobile divider technique and its related inspection procedure, which was first proposed and used in Calabria (Southern Italy) from 2014 to 2016. We report preliminary data concerning the number of detected SHBs and the time spent for their detection, based on the inspection of two apiaries in Calabria region, using both methods (official method and mobile divider method). The preliminary data presented here show that, on average, the mobile divider method seems to be able to recover a slightly higher number of beetles (0.9 adults) per inspected hive, in a shorter time (25 s).

New Matrixes to Diagnose Aethina Tumida Presence at Apiary Level

Aethina tumida (Coleoptera: Nitidulidae - Small Hive Beetle - SHB), is a parasite of honey bee colonies that causes the notifiable disease called aethinosis. In 2014, SHB was detected in Southern Italy, where it is still present (Calabria region). As part of surveillance activities, official diagnosis of the disease is performed by veterinary services via visual inspection of single hives in the apiaries. New outbreaks can be eradicated and the spread of SHBs limited by early detection of new introductions. We report an alternative protocol for the diagnosis of SHB through swabs and hive debris analysis by PCR. This was tested in three apiaries through the evaluation of different SHB infestation levels with a hive inspection method. This approach for sampling, followed by biomolecular methods, was fast and useful in unfavorable conditions (bad weather, aggressiveness, robbing), could be integrated in the official diagnosis procedures and may act as pre-clinic indicator.

Study on Major Parasitic Diseases of Adult Honeybees in Three Districts of Kaffa Zone, Southern Ethiopia

Background and Methods. Since regular and timely control of all factors that endanger honeybee life and threaten their products is needed for effective apiculture, cross-sectional research was performed in the three districts (Bonga, Chena, and Gimbo) of the Kaffa Zone to assess the prevalence of major honeybee diseases. A total of 384 honeybee samples from 15 operating apiaries were chosen from each kebele based on their ability to provide a bee sample and the apiary farm’s accessibility. From each district, 128 bee colonies were selected from different apiaries; of these, there were 209 transitional, 74 traditional, and 101 modern hives. Results. Out of 384 samples, the overall prevalence of small hive beetle, Nosema apis, and amoeba (Malpighamoeba mellificae) diseases was 39%, 45.3%, and 40.3%, respectively. The overall prevalence of these three diseases was ascertained, with a statistically significant ( p < 0.05 ) variation in the overall prevalence of N. apis disease and amoeba (M. mellificae) disease between hive type and agroecologies. Conclusion. The research indicates that N. apis, M. mellificae, and small hive beetle are the major parasites that affect honeybees in the study area. In order to classify and describe honeybee diseases and pests associated with Ethiopia’s local honeybees, several field diagnostic surveys and laboratory research works need to be performed.