Super DFM HoneyBee - Direct Fed Microbials

Super DFM HoneyBee – Blind Study – 2014
Goal: investigate impact of direct-fed microbial product, Super DFM Honeybee, on honeybee gut
microflora.
Summary: Supplementation with direct-fed microbial product, Super DFM Honeybee, significantly
increases Lactobacillus and Bacillus populations in honeybee gut. Terramycin treatment decreases
Lactobacillus populations in honeybee gut.
Objective 1. Lactobacillus are a natural constituent of commensal microflora of honeybees, Apis
mellifera. However, pathological and drug-induced changes in microflora lead to a decrease in natural
prevalence of the Lactobacillus. Supplementation with direct-fed microbial product, Super DFM
Honeybee, is expected to result in significant increase in Lactobacillus population in honeybee gut. Gut
Lactobacillus and other commensal bacterial play a role in inhibiting honeybee pathogens and therefore
play a critical role in honeybee health.
Result. We are able to demonstrate a significant increase in Lactobacillus population in the guts of
honeybees fed Super DFM Honeybee. Compared to untreated honeybee control group, honeybees fed
SuperDFM showed 9-fold increase in Lactobacillus population in the gut (p-value = 0.03).
2.00E+06
LAB concentration, CFU/g
1.80E+06
1.60E+06
1.40E+06
1.20E+06
1.00E+06
8.00E+05
6.00E+05
4.00E+05
2.00E+05
0.00E+00
negative control
SDFM
Objective 2. Antibiotic treatment (terramycin) negatively impacts commensal microflora of honeybees.
Lactobacillus population in honeybee gut is expected to decrease after terramycin treatment.
Result. We were able to quantitate 10 fold decrease in Lactobacillus population after terramycin
treatment.
6.00E+06
LAB concentration, CFU/g
5.00E+06
4.00E+06
3.00E+06
2.00E+06
1.00E+06
0.00E+00
negative control
SDFM - before
SDFM - after terramycin
terramycin treatment
treatment
Objective 3. Optimal concentration of probiotic supplementation was tested.
Result. Our data indicate that increasing Lactobacillus concentration in probiotic supplementation 100
fold did not proportionately increase concentration of Lactobacillus in honeybee gut, suggesting that
Lactobacillus in honeybee gut can reach a finite capacity. No adverse honeybee health effects were
observed when probiotic supplementation was increased 100 fold.
8.00E+06
LAB concentration, CFU/g
7.00E+06
6.00E+06
5.00E+06
4.00E+06
3.00E+06
2.00E+06
1.00E+06
0.00E+00
SDFM control
SDFM high dose
Objective 4. Analyze colonization of honeybee gut with beneficial Bacillus species.
Result. Besides Lactobacillus, we detected proprietary Bacillus species included in formulation of Super
DFM honeybee in the guts of all tested honeybee populations that were supplemented with SuperDFM.
% samples positive for Bacillus
100
90
80
70
60
50
40
30
20
10
0
negative control
SDFM treated
Methods.
Honeybee maintenance.
Honeybee microbial analysis. Live honeybees were shipped to the laboratory (Figure 1) and euthanized
by placing them into -20 degrees Celsius freezer. 1 gram of honeybees (approximately 3 honeybees)
were homogenized in 1 milliliter of 1% peptone water, and 10 fold dilutions of gut contents in peptone
were made (Figure 2). Dilutions were plated on MRS media for Lactobacillus quantification and TSA
medium for Bacillus quantification.
Figure 1.
Figure 2.

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