Effects of Nosema ceranae (Dissociodihaplophasida: Nosematidae) and Flupyradifurone on Olfactory Learning in Honey Bees, Apis mellifera (Hymenoptera: Apidae)

Abstract The health of insect pollinators, particularly the honey bee, Apis mellifera (Linnaeus, 1758), is a major concern for agriculture and ecosystem health. In response to mounting evidence supporting the detrimental effects of neonicotinoid pesticides on pollinators, a novel ‘bee safe’ butenolide compound, flupyradifurone (FPF) has been registered for use in agricultural use. Although FPF is not a neonicotinoid, like neonicotinoids, it is an excitotoxic nicotinic acetylcholine receptor agonist. In addition, A. mellifera faces threats from pathogens, such as the microsporidian endoparasite, Nosema ceranae (Fries et al. 1996). We therefore sought 1) to increase our understanding of the potential effects of FPF on honey bees by focusing on a crucial behavior, the ability to learn and remember an odor associated with a food reward, and 2) to test for a potential synergistic effect on such learning by exposure to FPF and infection with N. ceranae. We found little evidence that FPF significantly alters learning and memory at short-term field-realistic doses. However, at high doses and at chronic, field-realistic exposure, FPF did reduce learning and memory in an olfactory conditioning task. Infection with N. ceranae also reduced learning, but there was no synergy (no significant interaction) between N. ceranae and exposure to FPF. These results suggest the importance of continued studies on the chronic effects of FPF.

Download Full-text

Nectar non-protein amino acids (NPAAs) do not change nectar palatability but enhance learning and memory in honey bees

Scientific Reports ◽

10.1038/s41598-021-90895-z ◽

2021 ◽

Vol 11 (1) ◽

Author(s):

Daniele Carlesso ◽

Stefania Smargiassi ◽

Elisa Pasquini ◽

Giacomo Bertelli ◽

David Baracchi

Keyword(s):

Amino Acids ◽

Learning And Memory ◽

Honey Bees ◽

Feeding Preference ◽

Memory Retention ◽

Floral Nectar ◽

Olfactory Conditioning ◽

Specific Memory ◽

Protein Amino Acids ◽

Valuable Compounds

AbstractFloral nectar is a pivotal element of the intimate relationship between plants and pollinators. Nectars are composed of a plethora of nutritionally valuable compounds but also hundreds of secondary metabolites (SMs) whose function remains elusive. Here we performed a set of behavioural experiments to study whether five ubiquitous nectar non-protein amino acids (NPAAs: β-alanine, GABA, citrulline, ornithine and taurine) interact with gustation, feeding preference, and learning and memory in Apis mellifera. We showed that foragers were unable to discriminate NPAAs from water when only accessing antennal chemo-tactile information and that freely moving bees did not exhibit innate feeding preferences for NPAAs. Also, NPAAs did not alter food consumption or longevity in caged bees over 10 days. Taken together our data suggest that natural concentrations of NPAAs did not alter nectar palatability to bees. Olfactory conditioning assays showed that honey bees were more likely to learn a scent when it signalled a sucrose reward containing either β-alanine or GABA, and that GABA enhanced specific memory retention. Conversely, when ingested two hours prior to conditioning, GABA, β-alanine, and taurine weakened bees’ acquisition performances but not specific memory retention, which was enhanced in the case of β-alanine and taurine. Neither citrulline nor ornithine affected learning and memory. NPAAs in nectars may represent a cooperative strategy adopted by plants to attract beneficial pollinators.

Download Full-text

Effects of Prebiotics and Probiotics on Honey Bees (Apis mellifera) Infected with the Microsporidian Parasite Nosema ceranae

Microorganisms ◽

10.3390/microorganisms9030481 ◽

2021 ◽

Vol 9 (3) ◽

pp. 481

Author(s):

Daniel Borges ◽

Ernesto Guzman-Novoa ◽

Paul H. Goodwin

Keyword(s):

Apis Mellifera ◽

Honey Bees ◽

Bifidobacterium Bifidum ◽

Nosema Ceranae ◽

Food Supplementation ◽

Microsporidian Parasite ◽

Single Strain ◽

Acacia Gum ◽

Bee Health ◽

Commercial Probiotics

Nosema ceranae is a microsporidian fungus that parasitizes the midgut epithelial cells of honey bees, Apis mellifera. Due to the role that midgut microorganisms play in bee health and immunity, food supplementation with prebiotics and probiotics may assist in the control of N. ceranae. The dietary fiber prebiotics acacia gum, inulin, and fructooligosaccharides, as well as the commercial probiotics Vetafarm Probotic, Protexin Concentrate single-strain (Enterococcus faecium), and Protexin Concentrate multi-strain (Lactobacillus acidophilus, L. plantarum, L. rhamnosus, L. delbrueckii, Bifidobacterium bifidum, Streptococcus salivarius, and E. faecium) were tested for their effect on N. ceranae spore loads and honey bee survivorship. Bees kept in cages were inoculated with N. ceranae spores and single-dose treatments were administered in sugar syrup. Acacia gum caused the greatest reduction in N. ceranae spore numbers (67%) but also significantly increased bee mortality (62.2%). However, Protexin Concentrate single-strain gave similarly reduced spore numbers (59%) without affecting the mortality. In a second experiment, multiple doses of the probiotics revealed significantly reduced spore numbers with 2.50 mg/mL Vetafarm Probotic, and 0.25, 1.25, and 2.50 mg/mL Protexin Concentrate single-strain. Mortality was also significantly reduced with 1.25 mg/mL Protexin Concentrate single-strain. N. ceranae-inoculated bees fed 3.75 mg/mL Vetafarm Probotic had higher survival than N. ceranae-inoculated bees, which was similar to that of non-inoculated bees, while N. ceranae-inoculated bees fed 2.50 mg/mL Protexin Concentrate single-strain, had significantly higher survival than both N. ceranae-inoculated and non-inoculated bees. Protexin Concentrate single-strain is promising as it can reduce N. ceranae proliferation and increase bee survivorship of infected bees, even compared to healthy, non-infected bees.

Download Full-text

Nosema ceranae has been infecting honey bees Apis mellifera in Italy since at least 1993

Journal of Apicultural Research ◽

10.3896/ibra.1.52.2.11 ◽

2013 ◽

Vol 52 (2) ◽

pp. 60-61 ◽

Cited By ~ 13

Author(s):

Ezio Ferroglio ◽

Stefania Zanet ◽

Nancy Peraldo ◽

Elisa Tachis ◽

Anna Trisciuoglio ◽

...

Keyword(s):

Apis Mellifera ◽

Honey Bees ◽

Nosema Ceranae

Download Full-text

Permanent prevalence of Nosema ceranae in honey bees (Apis mellifera) in Hungary

Acta Veterinaria Hungarica ◽

10.1556/004.2015.034 ◽

2015 ◽

Vol 63 (3) ◽

pp. 358-369 ◽

Cited By ~ 1

Author(s):

Tamás Csáki ◽

Miklós Heltai ◽

Ferenc Markolt ◽

Balázs Kovács ◽

László Békési ◽

...

Keyword(s):

Apis Mellifera ◽

Honey Bees ◽

Mediterranean Climate ◽

Laboratory Method ◽

Seasonal Distribution ◽

Nosema Ceranae ◽

Infection Rates ◽

Infection Level ◽

Spore Count ◽

The Mean

Nosema ceranae is present in honey bee (Apis mellifera L.) colonies worldwide. Studies on the comparative virulence of N. ceranae and N. apis showed significant differences in individual mortality, and the prevalence of N. ceranae seems to be predominant in both the continental and the Mediterranean climate regions. This study attempted to monitor the geographical and seasonal distribution of these two Nosema species in Hungary, using a simple laboratory method. The distribution of N. ceranae and N. apis infection rates along all seasons was homogeneous (P = 0.57). In co-infected samples, the intensity of N. ceranae infection was always significantly higher than that of N. apis infection (P < 0.001). The infection rate of infected bees in exterior samples was higher than in interior samples in each season; however, the differences were not statistically significant. The species N. ceranae had been present in Hungary already in 2004. Statistical analysis of data shows that the infection level is best represented by sampling exterior bees to establish the proportion of infected bees rather than by determining the mean spore count.

Download Full-text

Age and Method of Inoculation Influence the Infection of Worker Honey Bees (Apis mellifera) by Nosema ceranae

Insects ◽

10.3390/insects10120417 ◽

2019 ◽

Vol 10 (12) ◽

pp. 417 ◽

Cited By ~ 2

Author(s):

Almudena Urbieta-Magro ◽

Mariano Higes ◽

Aránzazu Meana ◽

Laura Barrios ◽

Raquel Martín-Hernández

Keyword(s):

Apis Mellifera ◽

Honey Bees ◽

Parasite Load ◽

Nosema Ceranae ◽

Microsporidian Parasite ◽

Age Cohort ◽

Group A ◽

Time Of Infection ◽

Group B ◽

The Individual

The microsporidian parasite Nosema ceranae is a highly prevalent, global honey bee pathogen. Apis mellifera is considered to be a relatively recent host for this microsporidia, which raises questions as to how it affects its host’s physiology, behavior and longevity, both at the individual and colony level. As such, honey bees were inoculated with fresh purified spores of this pathogen, both individually (Group A) or collectively (Group B) and they were studied from 0 to 15 days post-emergence (p.e.) to evaluate the effect of bee age and the method of inoculation at 7 days post-infection. The level of infection was analyzed individually by qPCR by measuring the relative amount of the N. ceranae polar tubule protein 3 (PTP3) gene. The results show that the bee’s age and the method of infection directly influence parasite load, and thus, early disease development. Significant differences were found regarding bee age at the time of infection, whereby the youngest bees (new-born and 1 day p.e.) developed the highest parasite load, with this load decreasing dramatically in bees infected at 2 days p.e. before increasing again in bees infected at 3–4 days p.e. The parasite load in bees infected when older than 4 days p.e. diminished as they aged. When the age cohort data was pooled and grouped according to the method of infection, a significantly higher mean concentration and lower variation in N. ceranae infection was evident in Group A, indicating greater variation in experimental infection when spores were administered collectively to bees through their food. In summary, these data indicate that both biological and experimental factors should be taken into consideration when comparing data published in the literature.

Download Full-text

Changes with age in olfactory conditioning performance of worker honey bees (Apis mellifera)

Apidologie ◽

10.1051/apido:2001125 ◽

2001 ◽

Vol 32 (3) ◽

pp. 231-242 ◽

Cited By ~ 20

Author(s):

David Laloi ◽

Marie Gallois ◽

Bernard Roger ◽

Minh-H� Pham-Del�gue

Keyword(s):

Apis Mellifera ◽

Honey Bees ◽

Olfactory Conditioning ◽

Conditioning Performance

Download Full-text

Brain transcriptomes of honey bees ( Apis mellifera ) experimentally infected by two pathogens: Black queen cell virus and Nosema ceranae

Genomics Data ◽

10.1016/j.gdata.2016.09.010 ◽

2016 ◽

Vol 10 ◽

pp. 79-82 ◽

Cited By ~ 9

Author(s):

Vincent Doublet ◽

Robert J. Paxton ◽

Cynthia M. McDonnell ◽

Emeric Dubois ◽

Sabine Nidelet ◽

...

Keyword(s):

Apis Mellifera ◽

Honey Bees ◽

Nosema Ceranae ◽

Black Queen Cell Virus ◽

Queen Cell

Download Full-text

Does fumagillin control the recently detected invasive parasite Nosema ceranae in western honey bees (Apis mellifera)?

Journal of Invertebrate Pathology ◽

10.1016/j.jip.2008.04.005 ◽

2008 ◽

Vol 99 (3) ◽

pp. 342-344 ◽

Cited By ~ 77

Author(s):

Geoffrey R. Williams ◽

Michelle A. Sampson ◽

Dave Shutler ◽

Richard E.L. Rogers

Keyword(s):

Apis Mellifera ◽

Honey Bees ◽

Nosema Ceranae

Download Full-text

Nosemosis control in European honey bees, Apis mellifera, by silencing the gene encoding Nosema ceranae polar tube protein 3

Journal of Experimental Biology ◽

10.1242/jeb.184606 ◽

2018 ◽

Vol 221 (19) ◽

pp. jeb184606 ◽

Cited By ~ 6

Author(s):

Cristina Rodríguez-García ◽

Jay D. Evans ◽

Wenfeng Li ◽

Belén Branchiccela ◽

Jiang Hong Li ◽

...

Keyword(s):

Apis Mellifera ◽

Honey Bees ◽

Nosema Ceranae ◽

Polar Tube ◽

Gene Encoding

Download Full-text

Silencing the Honey Bee (Apis mellifera) Naked Cuticle Gene (nkd) Improves Host Immune Function and Reduces Nosema ceranae Infections

Applied and Environmental Microbiology ◽

10.1128/aem.02105-16 ◽

2016 ◽

Vol 82 (22) ◽

pp. 6779-6787 ◽

Cited By ~ 22

Author(s):

Wenfeng Li ◽

Jay D. Evans ◽

Qiang Huang ◽

Cristina Rodríguez-García ◽

Jie Liu ◽

...

Keyword(s):

Apis Mellifera ◽

Immune Function ◽

Honey Bee ◽

Honey Bees ◽

Nosema Ceranae ◽

Negative Regulator ◽

Mrna Levels ◽

Content Type ◽

Bee Disease

ABSTRACTNosema ceranaeis a new and emerging microsporidian parasite of European honey bees,Apis mellifera, that has been implicated in colony losses worldwide. RNA interference (RNAi), a posttranscriptional gene silencing mechanism, has emerged as a potent and specific strategy for controlling infections of parasites and pathogens in honey bees. While previous studies have focused on the silencing of parasite/pathogen virulence factors, we explore here the possibility of silencing a host factor as a mechanism for reducing parasite load. Specifically, we used an RNAi strategy to reduce the expression of a honey bee gene,naked cuticle(nkd), which is a negative regulator of host immune function. Our studies found thatnkdmRNA levels in adult bees were upregulated byN. ceranaeinfection (and thus, the parasite may use this mechanism to suppress host immune function) and that ingestion of double-stranded RNA (dsRNA) specific tonkdefficiently silenced its expression. Furthermore, we found that RNAi-mediated knockdown ofnkdtranscripts inNosema-infected bees resulted in upregulation of the expression of several immune genes (Abaecin,Apidaecin,Defensin-1, andPGRP-S2), reduction ofNosemaspore loads, and extension of honey bee life span. The results of our studies clearly indicate that silencing the hostnkdgene can activate honey bee immune responses, suppress the reproduction ofN. ceranae, and improve the overall health of honey bees. This study represents a novel host-derived therapeutic for honey bee disease treatment that merits further exploration.IMPORTANCEGiven the critical role of honey bees in the pollination of agricultural crops, it is urgent to develop strategies to prevent the colony decline induced by the infection of parasites/pathogens. Targeting parasites and pathogens directly by RNAi has been proven to be useful for controlling infections in honey bees, but little is known about the disease impacts of RNAi silencing of host factors. Here, we demonstrate that knocking down the honey bee immune repressor-encodingnkdgene can suppress the reproduction ofN. ceranaeand improve the overall health of honey bees, which highlights the potential role of host-derived and RNAi-based therapeutics in controlling the infections in honey bees. The information obtained from this study will have positive implications for honey bee disease management practices.

Download Full-text