Lactic acid bacteria (LAB) are well recognized beneficial host-associated members of the microbiota of humans and animals. Yet LAB-associations of invertebrates have been poorly characterized and their functions remain obscure. Here we show that honeybees possess an abundant, diverse and ancient LAB microbiota in their honey crop with beneficial effects for bee health, defending them against microbial threats. Our studies of LAB in all extant honeybee species plus related apid bees reveal one of the largest collections of novel species from the genera Lactobacillus and Bifidobacterium ever discovered within a single insect and suggest a long (>80 mya) history of association. Bee associated microbiotas highlight Lactobacillus kunkeei as the dominant LAB member. Those showing potent antimicrobial properties are acquired by callow honey bee workers from nestmates and maintained within the crop in biofilms, though beekeeping management practices can negatively impact this microbiota. Prophylactic practices that enhance LAB, or supplementary feeding of LAB, may serve in integrated approaches to sustainable pollinator service provision. We anticipate this microbiota will become central to studies on honeybee health, including colony collapse disorder, and act as an exemplar case of insect-microbe symbiosis.

A polyphasic taxonomic study of the two subspecies of Paenibacillus larvae, Paenibacillus larvae subsp. larvae and Paenibacillus larvae subsp. pulvifaciens, supported the reclassification of the subspecies into one species, Paenibacillus larvae, without subspecies separation. Our conclusions are based on the analysis of six reference strains of P. larvae subsp. pulvifaciens and three reference strains and 44 field isolates of P. larvae. subsp. larvae. The latter originated from brood or honey of clinically diseased honey bee colonies or from honey of both clinically diseased and asymptomatic colonies from Sweden, Finland and Germany. Colony and spore morphology, as well as the metabolism of mannitol and salicin, did not allow a clear identification of the two subspecies and SDS-PAGE of whole-cell proteins did not support the subspecies differentiation. For genomic fingerprinting, repetitive element-PCR fingerprinting using ERIC primers and PFGE of bacterial DNA were performed. The latter method is a high-resolution DNA fingerprinting method proven to be superior to most other methods for biochemical and molecular typing and has not previously been used to characterize P. larvae. ERIC-PCR identified four different genotypes, while PFGE revealed two main clusters. One cluster included most of the P. larvae subsp. larvae field isolates, as well as all P. larvae subsp. pulvifaciens reference strains. The other cluster comprised the pigmented variants of P. larvae subsp. larvae. 16S rRNA gene sequences were determined for some strains. Finally, exposure bioassays demonstrated that reference strains of P. larvae subsp. pulvifaciens were pathogenic for honey bee larvae, producing symptoms similar to reference strains of P. larvae subsp. larvae. In comparison with the type strain for P. larvae subsp. larvae, ATCC 9545 T , the P. larvae subsp. pulvifaciens strains tested were even more virulent, since they showed a shorter LT 100 . An emended description of the species is given.

We evaluated the antagonistic effects of newly identified lactic acid bacteria (LAB) in the genera Lactobacillus and Bifidobacterium, originating from the honey stomach, on the honey bee pathogen, Paenibacillus larvae. We used inhibition assays on agar plates and honey bee larval bioassays to investigate the effects of honey bee LAB on P. larvae growth in vitro and on AFB infection in vivo. The individual LAB phylotypes showed different inhibition properties against P. larvae growth on agar plates, whereas a combination of all eleven LAB phylotypes resulted in a total inhibition (no visible growth) of P. larvae. Adding the LAB mixture to the larval food significantly reduced the number of AFB infected larvae in exposure bioassays. The results demonstrate that honey bee specific LAB possess beneficial properties for honey bee health. Possible benefits to honey bee health by enhancing growth of LAB or by applying LAB to honey bee colonies should be further investigated.

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-The US National Honey Bee Disease Survey sampled colony pests and diseases from 2009 to 2014. We verified the absence of Tropilaelaps spp., the Asian honey bee (Apis cerana ), and slow bee paralysis virus. Endemic health threats were quantified, including Varroa destructor , Nosema spp., and eight honey bee viruses. Varroa loads varied across years, with annual fall peaks; Nosema peaked January to April. Migratory beekeepers had significantly lower Varroa prevalence (84.9 vs. 97.0 %) and loads (3.65 ± 0.28 vs. 5.99 ± 0.22) than stationary operations, while Nosema was more prevalent (59.9 vs. 46.7 %) in migratory colonies. Since 2010, chronic bee paralysis virus prevalence doubled annually. We detected strong positive relationships between V. destructor and Varroa -transmitted viruses, between Nosema and Lake Sinai virus 2, and a positive relationship across several viral pathogens of bees. The results provide a disease baseline to help identify drivers of poor bee health.Apis mellifera / disease survey / pests / parasite / USA

SummaryA variety of methods are used in honey bee research and differ depending on the level at which the research is conducted. On an individual level, the handling of individual honey bees, including the queen, larvae and pupae are required. There are different methods for the immobilising, killing and storing as well as determining individual weight of bees. The precise timing of developmental stages is also an important aspect of sampling individuals for experiments. In order to investigate and manipulate functional processes in honey bees, e.g. memory formation and retrieval and gene expression, microinjection is often used. A method that is used by both researchers and beekeepers is the marking of queens that serves not only to help to locate her during her life, but also enables the dating of queens. Creating multiple queen colonies allows the beekeeper to maintain spare queens, increase brood production or ask questions related to reproduction. On colony level, very useful techniques are the measurement of intra hive mortality using dead bee traps, weighing of full hives, collecting pollen and nectar, and digital monitoring of brood development via location recognition. At the population level, estimation of population density is essential to evaluate the health status and using beelines help to locate wild colonies. These methods, described in this paper, are especially valuable when investigating the effects of pesticide applications, environmental pollution and diseases on colony survival. Métodos estándar diversos para la investigación en Apis mellifera ResumenEn la investigación de la abeja de la miel, se han usado una variedad de métodos que se diferencian en función del nivel en el que se realiza la investigación. Al nivel individual, el manejo de las abejas individuales es necesario, incluyendo a la reina, las larvas y las pupas. Existen diferentes métodos para la inmovilización, mortandad y almacenamiento, así como para la determinación del peso individual de las abejas. La precisión en la sincronización de las etapas de desarrollo es también un aspecto importante de los experimentos con muestreos individuales.La microinyección se utiliza a menudo con el fin de investigar y manipular los procesos funcionales de las abejas melíferas, como por ejemplo, la formación y recuperación de la memoria y la expresión génica. Un método utilizado tanto por investigadores como apicultores es el marcado de las reinas, que sirve no sólo para ayudar a localizarlas durante su vida, sino que también permite su datación. La creación de varias colmenas a partir de reinas permite al apicultor mantener reinas de repuesto, aumentar la producción de cría o hacer preguntas relacionadas con la reproducción. Al nivel de colmena, la medición de la mortalidad intra colmena utilizando trampas de abejas muertas, el pesaje de las colmenas completas, la recolección de polen y néctar, y el seguimiento digital del desarrollo de la cría a través del reconocimiento de su ubicación, son algunas de las técnicas más útiles. Al nivel p...

The honey bee ectoparasitic mite, Varroa destructor, has a world-wide distribution and inflicts more damage than all other known apicultural diseases. However, Varroa-induced colony mortality is more accurately a result of secondary virus infections vectored by the mite. This means that honey bee resistance to Varroa may include resistance or tolerance to virus infections. The aim of this study was to see if this is the case for a unique population of mite-resistant (MR) European honey bees on the island of Gotland, Sweden. This population has survived uncontrolled mite infestation for over a decade, developing specific mite-related resistance traits to do so. Using RT-qPCR techniques, we monitored late season virus infections, Varroa mite infestation and honey bee colony population dynamics in the Gotland MR population and compared this to mite-susceptible (MS) colonies in a close by apiary. From summer to autumn the deformed wing virus (DWV) titres increased similarly between the MR and MS populations, while the black queen cell virus (BQCV) and sacbrood virus (SBV) titres decreased substantially in the MR population compared to the MS population by several orders of magnitude. The MR colonies all survived the following winter with high mite infestation, high DWV infection, small colony size and low proportions of autumn brood, while the MS colonies all perished. Possible explanations for these changes in virus titres and their relevance to Varroa resistance and colony winter survival are discussed.