scholarly journals Niche partitioning facilitates coexistence of closely related honey bee gut bacteria

eLife ◽  
2021 ◽  
Vol 10 ◽  
Author(s):  
Silvia Brochet ◽  
Andrew Quinn ◽  
Ruben AT Mars ◽  
Nicolas Neuschwander ◽  
Uwe Sauer ◽  
...  
Keyword(s):  
Honey Bee ◽  
Resource Partitioning ◽  
Niche Partitioning ◽  
Gut Bacteria ◽  
Host Association ◽  
Ecological Processes ◽  
Liquid Cultures ◽  
Bacterial Interactions

Ecological processes underlying bacterial coexistence in the gut are not well understood. Here, we disentangled the effect of the host and the diet on the coexistence of four closely related Lactobacillus species colonizing the honey bee gut. We serially passaged the four species through gnotobiotic bees and in liquid cultures in the presence of either pollen (bee diet) or simple sugars. Although the four species engaged in negative interactions, they were able to stably coexist, both in vivo and in vitro. However, coexistence was only possible in the presence of pollen, and not in simple sugars, independent of the environment. Using metatranscriptomics and metabolomics, we found that the four species utilize different pollen-derived carbohydrate substrates indicating resource partitioning as the basis of coexistence. Our results show that despite longstanding host association, gut bacterial interactions can be recapitulated in vitro providing insights about bacterial coexistence when combined with in vivo experiments.

scholarly journals Niche partitioning facilitates coexistence of closely related gut bacteria

2021 ◽  
Author(s):  
Silvia Brochet ◽  
Andrew Quinn ◽  
Ruben A.T. Mars ◽  
Nicolas Neuschwander ◽  
Uwe Sauer ◽  
...  
Keyword(s):  
Honey Bee ◽  
Resource Partitioning ◽  
Niche Partitioning ◽  
Gut Bacteria ◽  
Ecological Processes ◽  
Liquid Cultures ◽  
Symbiotic Associations ◽  
Bacterial Interactions

AbstractEcological processes underlying bacterial coexistence in the gut are not well understood. Here, we disentangled the effect of the host and the diet on the coexistence of four closely related Lactobacillus species colonizing the honey bee gut. We serially passaged the four species through gnotobiotic bees and in liquid cultures in the presence of either pollen (bee diet) or simple sugars. Although the four species engaged in negative interactions, they were able to stably coexist, both in vivo and in vitro. However, coexistence was only possible in the presence of pollen, but not in simple sugars independent of the environment. Using metatranscriptomics and metabolomics, we found that the four species utilize different pollen-derived carbohydrate substrates indicating resource partitioning as the basis of coexistence. Our results show that despite longstanding symbiotic associations, gut bacterial interactions can be recapitulated in vitro providing insights about bacterial coexistence when combined with in vivo experiments.

In vivo validation of in vitro quality tests for cryopreserved honey bee semen

Cryobiology ◽  
2012 ◽  
Vol 65 (2) ◽  
pp. 126-131 ◽  
Author(s):  
Jakob Wegener ◽  
Tanja May ◽  
Ulrich Knollmann ◽  
Günter Kamp ◽  
Karin Müller ◽  
...  
Keyword(s):  
Honey Bee ◽  
In Vivo Validation

In vitro antagonistic potential of gut bacteria isolated from indigenous honey bee race of Saudi Arabia against Paenibacillus larvae

2020 ◽  
Vol 59 (5) ◽  
pp. 825-833 ◽  
Author(s):  
Ahmad Al-Ghamdi ◽  
Amal Abdullah Al-Abbadi ◽  
Khalid Ali Khan ◽  
Hamed Ali Ghramh ◽  
Ashraf M. Ahmed ◽  
...  
Keyword(s):  
Saudi Arabia ◽  
Honey Bee ◽  
Gut Bacteria ◽  
Paenibacillus Larvae ◽  
Antagonistic Potential

Effect of inoculum age and physical parameters on in vitro culture of the entomopathogenic nematode Steinernema feltiae

2016 ◽  
Vol 91 (6) ◽  
pp. 686-695 ◽  
Author(s):  
L.G. Leite ◽  
D.I. Shapiro-Ilan ◽  
S. Hazir ◽  
M.A. Jackson
Keyword(s):  
Liquid Medium ◽  
Solid Medium ◽  
Entomopathogenic Nematode ◽  
Steinernema Feltiae ◽  
Physical Parameters ◽  
Symbiotic Association ◽  
Liquid Cultures ◽  
Inoculum Age

AbstractEntomopathogenic nematodes (EPNs) of the families Steinernematidae and Heterorhabditidae have a symbiotic association with bacteria which makes them virulent against insects. EPNs have been mass produced using in vivo and in vitro methods, including both solid and liquid fermentation. This study assessed the effect of nematode inoculum age on the production of Steinernema feltiae in liquid, solid and biphasic processes. Several physical parameters were also assessed: the effect of medium viscosity, flask size and aeration speed on the recovery and yield of infective juveniles (IJs). Inoculum age treatments included inoculum liquid cultures that were 7, 14, 21 and 28 days old. Nematodes from the same inoculum were added to one liquid medium (liquid culture), one solid medium with bacteria previously grown in sponge (solid culture) and a variation of the solid medium (a biphasic culture), in which the bacteria were first grown in liquid and, then, soaked into the sponges, with the purpose of providing a more homogeneous bacterial culture before nematode inoculation. Experiments were conducted in Erlenmeyer flasks. Eight treatments were established involving combinations of three variables: two media (with and without 0.2% agar), two flask sizes (250 and 150 ml) and two agitation speeds (180 and 280 rpm). The study showed increases in nematode yield for liquid cultures, but not for solid or biphasic cultures, with the advance of the inoculum age up to 28 days of growth. Furthermore, the addition of 0.2% agar to the liquid medium and increasing the aeration rate by using larger flasks with higher agitation speed may increase nematode recovery and final yield. The experiments were conducted using shake flasks but the results may also be applicable for bioreactors.

scholarly journals Revealing the in vivo growth and division patterns of mouse gut bacteria

2020 ◽  
Vol 6 (36) ◽  
pp. eabb2531
Author(s):  
Liyuan Lin ◽  
Qiuyue Wu ◽  
Jia Song ◽  
Yahui Du ◽  
Juan Gao ◽  
...  
Keyword(s):  
Amino Acid ◽  
Gut Microbiota ◽  
Growth Dynamics ◽  
Gut Bacteria ◽  
Metabolic Labeling ◽  
Taxonomic Identification ◽  
In Vivo Growth

Current techniques for studying gut microbiota are unable to answer some important microbiology questions, like how different bacteria grow and divide in the gut. We propose a method that integrates the use of sequential d-amino acid–based in vivo metabolic labeling with fluorescence in situ hybridization (FISH), for characterizing the growth and division patterns of gut bacteria. After sequentially administering two d-amino acid–based probes containing different fluorophores to mice by gavage, the resulting dual-labeled peptidoglycans provide temporal information on cell wall synthesis of gut bacteria. Following taxonomic identification with FISH probes, the growth and division patterns of the corresponding bacterial taxa, including species that cannot be cultured separately in vitro, are revealed. Our method offers a facile yet powerful tool for investigating the in vivo growth dynamics of the bacterial gut microbiota, which will advance our understanding of bacterial cytology and facilitate elucidation of the basic microbiology of this gut “dark matter.”

scholarly journals In vivo and in vitro infection dynamics of honey bee viruses

2016 ◽  
Vol 6 (1) ◽  
Author(s):  
Jimena Carrillo-Tripp ◽  
Adam G. Dolezal ◽  
Michael J. Goblirsch ◽  
W. Allen Miller ◽  
Amy L. Toth ◽  
...  
Keyword(s):  
Honey Bee ◽  
Infection Dynamics ◽  
Bee Viruses

scholarly journals Antagonistic Effect of Gut Bacteria in the Hybrid Carniolan Honey Bee, Apis Mellifera Carnica, Against Ascosphaera Apis, the Causal Organism of Chalkbrood Disease

2014 ◽  
Vol 58 (1) ◽  
pp. 17-27 ◽  
Author(s):  
Mohamed O. M. Omar ◽  
Adhm M. Moustafa ◽  
Mohammad J. Ansari ◽  
Abdelsalam M. Anwar ◽  
Bassam F. Fahmy ◽  
...  
Keyword(s):  
Biological Control ◽  
Apis Mellifera ◽  
Honey Bee ◽  
Antagonistic Activity ◽  
Bacterial Strains ◽  
Ascosphaera Apis ◽  
Causal Organism ◽  
Apis Mellifera Carnica

Abstract The objective of this study was to isolate and characterize bacterial strains associated with the gut of the hybrid Carniolan honey bee, Apis mellifera carnica, and to determine their in vitro and in vivo potential against Ascosphaera apis, the causal organism of chalkbrood disease, with the purpose of exploring feasible biological control. Six bacterial strains were isolated from healthy worker honey bees by culture-dependent methods. Six fungal strains (A3, A4, A7, A8, A9, and A15) of A. apis were isolated from larvae suffering from chalkbrood disease on Yeast-Glucose-Starch agar (YGPSA) medium. All bacteria were identified by a combination of morphology, Gram stain, and 16S rRNA sequence analysis, and fungal strains were identified by morphology and 5.8S rRNA. In vitro and in vivo inhibition assays were carried out to determine the ability of bacterial isolates to inhibit A. apis, the causal agent of chalkbrood disease. The analysis of 16S rRNA sequences revealed that four bacterial strains (B2, B4, B10, and B100) belong to Bacillus subtilis species, and two strains (P1 and P5) belong to Pseudomonas fluorescence. Significant differences in antagonistic activity of all bacterial strains were observed. B. subtilis isolate B2 showed the highest antagonistic activity, as measured by the inhibition zone against A. apis, followed by the P1 strain of P. fluorescence. SEM analysis also supports the antagonistic activity of these bacteria against A. apis. This study provides a theoretical basis for biological control of honey bee chalkbrood disease.

scholarly journals In Vivo Expression of the Mannose-Resistant Fimbriae of Photorhabdus temperata K122 during Insect Infection

2004 ◽  
Vol 186 (3) ◽  
pp. 611-622 ◽  
Author(s):  
L. M. Meslet-Cladiere ◽  
A. Pimenta ◽  
E. Duchaud ◽  
I. B. Holland ◽  
M. A. Blight
Keyword(s):  
Pathogenic Bacteria ◽  
Galleria Mellonella ◽  
Liquid Cultures ◽  
Reverse Transcription Pcr ◽  
Greater Wax Moth ◽  
The Family ◽  
Complex Regulation ◽  
Photorhabdus Temperata

ABSTRACT Photorhabdus temperata K122 is an entomopathogenic bacterium symbiotically associated with nematodes of the family Heterorhabditidae. Surface fimbriae are important for the colonization of many pathogenic bacteria, and here we report the nucleotide sequence and analysis of the expression of a 12-kbp fragment encoding the mannose-resistant fimbriae of P. temperata (mrf). The mrf gene cluster contains 11 genes with an organization similar to that of the mrp locus from Proteus mirabilis. mrfI (encoding a putative recombinase) and mrfA (encoding pilin), the first gene in an apparent operon of nine other genes, are expressed from divergent promoters. The mrfI-mrfA intergenic region contains inverted repeats flanking the mrfA promoter. This region was shown to be capable of inversion, consistent with an ON/OFF regulation of the operon. In in vitro liquid cultures, both orientations were detected. Nevertheless, when we analyzed the expression of all of the genes in the mrf locus by semiquantitative reverse transcription-PCR during infection of Galleria mellonella (greater wax moth) larvae, expression of mrfA was not detected until 25 h postinfection, preceding the death of the larvae at 32 h. In contrast, mrfJ (a putative inhibitor of flagellar synthesis) was expressed throughout infection. Expression of mrfI was also detected only late in infection (25 to 30 h), indicating a possible increase in inversion frequency at this stage. In both in vitro liquid cultures and in vivo larval infections, the distal genes of the operon were expressed at substantially lower levels than mrfA. These results indicate the complex regulation of the mrf cluster during infection.

scholarly journals A bacterial symbiont protects honey bees from fungal disease

2020 ◽  
Author(s):  
Delaney L. Miller ◽  
Eric A. Smith ◽  
Irene L. G. Newton
Keyword(s):  
Honey Bee ◽  
Fungal Pathogens ◽  
Polyketide Synthase ◽  
Opportunistic Infections ◽  
Gene Clusters ◽  
Type I ◽  
Plant Host ◽  
Colony Losses

Fungi are the leading cause of insect disease, contributing to the decline of wild and managed populations1,2. For ecologically and economically critical species, such as the European honey bee (Apis mellifera), the presence and prevalence of fungal pathogens can have far reaching consequences, endangering other species and threatening food security3,4,5. Our ability to address fungal epidemics and opportunistic infections is currently hampered by the limited number of antifungal therapies6,7. Novel antifungal treatments are frequently of bacterial origin and produced by defensive symbionts (bacteria that associate with an animal/plant host and protect against natural enemies 89. Here we examined the capacity of a honey bee-associated bacterium, Bombella apis, to suppress the growth of fungal pathogens and ultimately protect bee brood (larvae and pupae) from infection. Our results showed that strains of B. apis inhibit the growth of two insect fungal pathogens, Beauveria bassiana and Aspergillus flavus, in vitro. This phenotype was recapitulated in vivo; bee brood supplemented with B. apis were significantly less likely to be infected by A. flavus. Additionally, the presence of B. apis reduced sporulation of A. flavus in the few bees that were infected. Analyses of biosynthetic gene clusters across B. apis strains suggest antifungal production via a Type I polyketide synthase. Secreted metabolites from B. apis alone were sufficient to suppress fungal growth, supporting this hypothesis. Together, these data suggest that B. apis protects bee brood from fungal infection by the secretion of an antifungal metabolite. On the basis of this discovery, new antifungal treatments could be developed to mitigate honey bee colony losses, and, in the future, could address fungal epidemics in other species.

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