Prognostic value of using bee and hive debris samples for the detection of American foulbrood disease in honey bee colonies

Forsgren, Eva; Laugen, Ane T.

doi:10.1007/s13592-013-0225-6

Cited by 35 publications

(37 citation statements)

References 23 publications

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“…9. Each set of PCR assays included serial dilutions of DNA extracted from known concentrations of P. larvae spores (total microscopic count) as external standards, for relating the qPCR data to spore counts (Forsgren, Stevanovic, & Fries, 2008). 10.…”

Section: Isopropanolmentioning

confidence: 99%

Standard methods forApis melliferabeeswax research

Svečnjak

Chesson

Gallina

et al. 2019

Journal of Apicultural Research

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Due to its multifunctional and complex role in the honey bee colony functioning and health (construction material allowing food storage, brood rearing, thermoregulation, mediation in chemical and mechanical communication, substrate for pathogens, toxins and waste), Apis mellifera beeswax has been widely studied over the last five decades. This is supported by a comprehensive set of scientific reports covering different aspects of beeswax research. In this article, we present an overview of the methods for studying chemical, biological, constructional, and quality aspects of beeswax. We provide a detailed description of the methods for investigating wax scales, comb construction and growth pattern, cell properties, chemical composition of beeswax using different analytical tools, as well as the analytical procedures for provenancing beeswax and beeswax-derived compounds based on the hydrogen isotope ratio (IRMS). Along with classical physico-chemical and sensory analysis, we describe more precise and accurate methods for detection of adulterants in beeswax (GC-MS and FTIR-ATR). Moreover, we present methods for studying the influence of beeswax (comb foundation) adulteration on comb construction. Analytical protocols for determining the pesticide residues using different chromatographic and spectroscopic techniques are also described. As beeswax is an agent of high risk for the transmission of bee diseases, we present methods for detection of pathogens in beeswax. To ensure the reproducibility of experiments and results, we present best practice approaches and detailed protocols for all methods described, as well as their advantages and disadvantages.

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Section: Isopropanolmentioning

confidence: 99%

Standard methods forApis melliferabeeswax research

Svečnjak

Chesson

Gallina

et al. 2019

Journal of Apicultural Research

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“…Worker honeybee samples were homogenized and cultivated on MYPGP-agar plates as described by Forsgren and Laugen [38]. Bacterial colonies were confirmed by PCR as described previously [39].…”

Section: Cultivation Of P Larvaementioning

confidence: 99%

Feeding Honeybee Colonies with Honeybee-Specific Lactic Acid Bacteria (Hbs-LAB) Does Not Affect Colony-Level Hbs-LAB Composition or Paenibacillus larvae Spore Levels, Although American Foulbrood Affected Colonies Harbor a More Diverse Hbs-LAB Community

et al. 2019

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The main current methods for controlling American Foulbrood (AFB) in honeybees, caused by the bacterial pathogen Paenibacillus larvae, are enforced incineration or prophylactic antibiotic treatment, neither of which is fully satisfactory. This has led to an increased interest in the natural relationships between the pathogenic and mutualistic microorganisms of the honeybee microbiome, in particular, the antagonistic effects of Honeybee-Specific Lactic Acid Bacteria (hbs-LAB) against P. larvae. We investigated whether supplemental administration of these bacteria affected P. larvae infection at colony level over an entire flowering season. Over the season, the supplements affected neither colony-level hbs-LAB composition nor naturally subclinical or clinical P. larvae spore levels. The composition of hbs-LAB in colonies was, however, more diverse in apiaries with a history of clinical AFB, although this was also unrelated to P. larvae spore levels. During the experiments, we also showed that qPCR could detect a wider range of hbs-LAB, with higher specificity and sensitivity than mass spectrometry. Honeybee colonies are complex super-organisms where social immune defenses, natural homeostatic mechanisms, and microbiome diversity and function play a major role in disease resistance. This means that observations made at the individual bee level cannot be simply extrapolated to infer similar effects at colony level. Although individual laboratory larval assays have clearly demonstrated the antagonistic effects of hbs-LAB on P. larvae infection, the results from the experiments presented here indicate that direct conversion of such practice to colony-level administration of live hbs-LAB is not effective.

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“…Samples of diseased cells were tested in the laboratory to confirm the diagnoses. The spore levels (hereafter: Spores) were determined from samples of 100 adult worker bees, as described previously [32,38]. The raw data consisted of P. larvae colony forming units (CFU) and the data are presented as CFU per bee (see also Additional file 3).…”

Section: Colony Assessments Afb Scoring and Spore Countingmentioning

confidence: 99%

“…One way to address the risk of pre-clinical infectious colonies for epidemic spread at multiple scales (local, regional, national) is to determine this risk directly from P. larvae spore levels in material sampled from the colony, thereby unambiguously identifying all infectious colonies rather than just those presenting detectable symptoms. It has been shown that adult bees provide most reliable samples for relating P. larvae spore levels to AFB symptoms, superior to either colony debris or honey samples [28,[31][32][33]. The spore load of individual bees is positively correlated to the likelihood of clinical symptoms [31,34].…”

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confidence: 99%

American foulbrood in a honeybee colony: spore-symptom relationship and feedbacks between disease and colony development

2020

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Background: The most severe bacterial disease of honeybees is American foulbrood (AFB). The epidemiology of AFB is driven by the extreme spore resilience, the difficulty of bees to remove these spores, and the considerable incidence of undetected spore-producing colonies. The honeybee collective defence mechanisms and their feedback on colony development, which involves a division of labour at multiple levels of colony organization, are difficult to model. To better predict disease outbreaks we need to understand the feedback between colony development and disease progression within the colony. We therefore developed Bayesian models with data from forty AFB-diseased colonies monitored over an entire foraging season to (i) investigate the relationship between spore production and symptoms, (ii) disentangle the feedback loops between AFB epidemiology and natural colony development, and (iii) discuss whether larger insect societies promote or limit within-colony disease transmission. Results: Rather than identifying a fixed spore count threshold for clinical symptoms, we estimated the probabilities around the relationship between spore counts and symptoms, taking into account modulators such as brood amount/number of bees and time post infection. We identified a decrease over time in the bees-to-brood ratio related to disease development, which should ultimately induce colony collapse. Lastly, two contrasting theories predict that larger colonies could promote either higher (classical epidemiological SIR-model) or lower (increasing spatial nest segregation and more effective pathogen removal) disease prevalence. Conclusions: AFB followed the predictions of the SIR-model, partly because disease prevalence and brood removal are decoupled, with worker bees acting more as disease vectors, infecting new brood, than as agents of social immunity, by removing infected brood. We therefore established a direct link between disease prevalence and social group size for a eusocial insect. We furthermore provide a probabilistic description of the relationship between AFB spore counts and symptoms, and how disease development and colony strength over a season modulate this relationship. These results help to better understand disease development within honeybee colonies, provide important estimates for further epidemiological modelling, and gained important insights into the optimal sampling strategy for practical beekeeping and honeybee research.

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Prognostic value of using bee and hive debris samples for the detection of American foulbrood disease in honey bee colonies

Cited by 35 publications

References 23 publications

Standard methods forApis melliferabeeswax research

Standard methods forApis melliferabeeswax research

Feeding Honeybee Colonies with Honeybee-Specific Lactic Acid Bacteria (Hbs-LAB) Does Not Affect Colony-Level Hbs-LAB Composition or Paenibacillus larvae Spore Levels, Although American Foulbrood Affected Colonies Harbor a More Diverse Hbs-LAB Community

American foulbrood in a honeybee colony: spore-symptom relationship and feedbacks between disease and colony development

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