Lactobacilli Symbiosis

This Blog contains information on how to use natural probiotics for honeybee health for those interested in the scientific part about probiotics and for the lay person who is not interested in the science. If you want just the bare facts and how to use them then scroll down to Formula 1 for the Lacto Formula and Formula 2 for the Honey Formula.

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Over the years of doing research on nearly everything I think about, I am always amazed at how the Universe never does anything singularly. Everything works together synergistically or through symbiosis. Nothing works on its own without something else incorporating its effort and support or cleans up. We breathe carbon dioxide and trees inhale it. We put out pollutants and Mother uses her breath, the wind, and moves the toxins through the trees and other green plants to cleanse it. The rain is recycled up from the ground by the sun drawing the moisture up through to make clouds only to fall back down again as rain or snow.

All living elements work through symbiosis in healing – nothing works alone. I.E., as a Master Herbalist, I learned that a good herbal formula contains 3 components: one to do the task, one to move the cleansing out of the body, and one to soothe behind. This is why Marshmallow Root and Slippery Elm Bark Powder are often used as they act as emollients as they move through the body.

In this blog I wish to talk about the symbiotic relationship in a honeybee’s microbiome gut and how it relates to our gut and other animals. Microbiome systems exist in everything. From the internet dictionary the microbiome is defined as: “…The microorganisms in a particular environment (including the body or a part of the body).

“We depend on a vast army of microbes to stay alive: a microbiome that protects us against germs, breaks down food to release energy, and produces vitamins.

  • The combined genetic material of the microorganisms in a particular environment.”

There are about 100 trillion bacteria in the human[i] gut. Both species have these bacteria[ii] that work similarly with good and bad bacteria in constant battle with each other with the good taking care of our immune system and the bad bacteria trying to undermine it. Mother has this duality in everything to keep our bodies and, hopefully, our intellect aware of the fact that we need both to appreciate both. Both are needed for us to learn what will heal and what will harm and what we need as a cure.

I have read that there are as many as 186 different bacteria in a honeybee’s[iii] gut.

Additionally, several studies hypothesize that the foregut (crop), a key interface between the pollination environment and hive food stores, contains a set of 13 lactic acid bacteria (LAB) that inoculate collected pollen and act in synergy to preserve pollen stores.

As a beekeeper I do a lot of research for my bees and have learned the importance of Lactic Acid Bacteria (LAB) in all living things, especially in our food. Lactobacilli is a LAB, as is Biobifidobacterium, and both are part of the microbiome in our guts as well as in the honeybee. Not one bacterium will do the job of healing. We need many bacteria to work together to heal.

Lactobacilli are in every living thing on the earth, including the soil, and can protect nearly all living elements from diseases and maintain good immune systems. Roundup is an herbicide that contains glyphosate and kills all the good bacteria that comes in contact with it. When sprayed this herbicide floats on the air for miles and miles until it rains before sinking into the soil. The good Lactobacilli bacteria in the soil prevents the Clostridium botulinum from coming up into the plant. With the lactobacilli dead in the soil, the Clostridium botulinum comes through our food as salmonella, e-coli, and listeria, just a few of the bad bacteria in the botulinum family.

I live in a GMO farming community where the farmer uses Roundup on a regular basis. I also have a neighbor who sprays the herbicide under the fence that holds his sheep. Whenever anyone uses herbicides, my bees develop European Foulbrood Disease (EFB). This link[iv] is a good website with pictures of EFB. When bees bring a chemical into the hive that is on their feet and bodies, they walk around on the comb, work the honey, etc. When something unnatural has been taken into the hive and things change within the hive, they get stressed and develop EFB, a bacterium called Melissococcus plutonius[v], is a LAB. EFB can be killed immediately with my lacto formula sprayed on the comb, and will protect the honeybee when the formula is put into their feed.

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Below is an excerpt from my August, 2016 Newsletter containing the Lacto Water formula and the formula to create your own lactobacilli from the bees own honey.

“Every living element on the earth – the soil, humans, animals, insects, fish – everything, need Lactobacillus and Biobifidobacterium for optimum health, and this includes honeybees.[vi] Honey is the bee’s prebiotic while the beebread is their probiotic; but both contain these bacteria, along with yeasts, vitamins, minerals, enzymes, etc. A probiotic is a product fermented by lactic acid that actually stimulates an immune response in the honeybee – as well as in humans. Most of these bacteria grow between 5.0 and 7.0 pH – 7.0 being mid-point. Anything lower is acidic. Anything higher is alkaline.[vii] All these bacteria will grow between 94° – 97° F – the same temperature of the human body and honeybee hive.[viii]

The honeybee stomach can contain upwards of 186 different bacterial strains.[ix] All of the strains grow in different pH environments. One bacterium in particular, Lactobacillus plantarum is one of two bacteria that studies show will kill American Foulbrood (AFB).[x],  The other bacterium that kills both AFB and EFB is L. kunkeei, which grows at 4.5 – 6.8 pH and prefers fructose, is contained in wine, flowers, and honey. The honeybee gut carries both the plantarum and the kunkeei bacteria in its stomach. The curious natural phenomenon about the plantarum bacteria is that it acidifies a growth medium to a low 2.0 – 4.0 pH within several hours.[xi]

Physical Requirements[xii]

  1. pH: Organisms can be classified as:

  2. Acidophiles: “Acid loving”.

u Grow at very low pH (0.1 to 5.4)

u Lactobacillus produces lactic acid, tolerates mild acidity.

Other bacteria fermented by the lactic acid also help with the honeybee’s immune system.

“The use of Lactobacillus species as probiotics has been found to enhance the immunity of honeybees, helping them to survive against the effect of pathogens and bring advantageous properties for honeybee health (Evans and Lopez, 2004; Forsgren et al., 2010).”[xiii]

When I first used the 8-strain, 30 ppb, to create my lacto water for my bees when they contracted EFB, they actively picked up. But they became very active when I added the 10-strain that included the L. casei[xiv]. These bacteria will also grow in just a few hours. Below is one of the research studies I used to create my Lacto water formula for EFB:

“Also, in beekeeping management, there are commercial diet supplements which contain probiotics and/or prebiotics. One such supplement recommended for the feeding of honeybees and other animals contains bacteria such as Lactobacillus casei, Lactobacillus plantarum, Rhodopseudomonas palustris, and yeast Saccharomyces cerevisiae. A further example, in addition to lactic acid bacteria (Lactobacillus acidophilus or L. casei) and Bifidobacterium lactis, also comprises prebiotics (Pătruică and Mot 2012; Pătruică and Hutu 2013; Andrearczyk et al. 2014).”[xv]

When you make sugar water solution with your own water, assuming it is at 7.0, and you don’t bring the pH down, mold will grow. Dr. Don Huber told me that Apple Cider Vinegar (ACV) would bring the pH down to 7.0 to equal honey’s p. Honey is capped at about 4.0 – 4.3 pH. Adding 1 Tablespoon of ACV to 1 gallon of sugar feed brought will bring the pH down to about 4.5 – 5.0, which would be the mean pH for good bacterial growth. I have heard of folks using distilled vinegar because it doesn’t attract wasps. This should be an indicator to you. Bees are trusting and will go to the distilled vinegar, just as they go to a swimming pool for the aspartame[xvi]

Since honey starts to ferment as soon as it is combined with water, my thought was to add 1 pint of honey to less than a gallon of sugar water so that some of the lactic acids and Biobifidobacterium already in the honey would grow. I would add 1 Tbsp of ACV to bring the pH down and allow other bacteria to grow – with the hypothesis that the L. plantarum would also grow and lower the pH more as it continues to multiply. The lower pH will grow the L. Kunkeei.

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The 2 Lacto Formulas

When the bees encounter glyphosate, during flight or while on plants, they may develop EFB, a disease brought on by stress. For the past 4 years when the farmers do major sprays (once before planting and again when the plants are about a foot high) I feed my bees my lacto formula. If I know my neighbors are spraying under their sheep fence, I feed my bees the formula. This formula has never failed me and works 100%. If the weather is fair and dry, it will take about 2-3 weeks to totally kill the disease. If it is damp and rainy, it will probably take about 3-5 weeks. If you catch EFB immediately, usually 1 or 2 sprays on the comb and 1 feed will work.

I DO NOT SUGGEST YOU USE THE STORE BOUGHT PROBIOTIC IF YOU USE HONEY. JUST LIKE YOU, TOO MUCH PROBIOTICS AND BEES MAY DEVELOP DIAAHREA.

You may want to get your own test strips. It is more accurate than those you would use to test their saliva. I get the strips off Amazon that measure the pH range of 0-13.[xvii]

Formula 1 – Lacto Water Using Only Probiotics

You can your probiotics from your Health Food Store. When I first made my Lacto Formula, I used the 8-strain until I learned about L casei. I switched to the Ultimate Probiotic 10-strain, 30 PPB, and discovered my bees enjoyed it more. So I stay with it. I use this probiotic for myself and take it to Africa with me as it is a freeze-dried in a blister packet and can take the heat. It hasn’t failed me. It contains Bifidobacterium lactis, B. breve, B. longum, Lactobacillus acidophilus, L. casaei, L. plantarum, L. paracasei, L. salivarius, L. rhamnosus, L. bulgarius

Step 1

In a cup of room temperature (tepid) water empty the contents of 1 probiotic capsule containing the 10 strains of Lactobacillus/Biobifidobacterium,30 ppb; stir and dissolve for a few minutes. Cover and refrigerate. Do not keep beyond 4-5 days.

Step 2

For 1:2 add 6 cups of white sugar to 1 gallon of water. For 1:1 add 11 cups of sugar to a gallon of water. Eyeballing it 1:1 is filling a glass (plastic emits a gas when hot) container about ¾ full of sugar. Put in enough hot water, not quite boiling, to dissolve the sugar (about ½ of a tea kettle). Stir often to dissolve the crystals. I do not use organic sugar as there are particles in the sugar that are not really digestible for the honeybee. When the sugar has dissolved, add enough cold water to fill the container to within 1” from the top. When totally cooled, add 2 Tablespoons of the Lacto water solution. Feed your bees.

EFB – Spraying the comb

In 1 pint of tepid water in a spray bottle, add 1 teaspoon of lacto water. Lightly spritz both sides of all frames, especially those filled with brood and larvae, and the sides of boxes in the hive body. Don’t need to spray bottom board or inner cover, unless they are solid, or the top outer cover. The EFB is affected immediately. I know this because I used an EFB test 10 minutes after I sprayed and came with a negative result, where it was positive before.

Repeat every 4 days for about 2-4 weeks until there is no longer any odor from the hive when you open it. 3 weeks is good. If there is a lot of rain or damp, humid climate it may take longer. You will notice the rotting smell will have lessened after the first use. If you catch the EFB in the early stages, say on 1 frame, you can eradicate the EFB in just 1 or 2 sprays. The larvae affected by EFB will dry up and the bees will carry it out of the cells.

You will need to feed the bees your sugar/lacto water formula and pollen during this time if there is not enough honey or bee bread in the hive. Bees are just like you when they are ill. They don’t want to do much and are not too functional outside the hive.

 

Formula 2 – Growing your own Lactobacillus using Honey

I wouldn’t suggest using nectar as it is not a complete product as honey. Once the honey is in the water, the lactic acid will start fermenting the solution. Honey also adds vitamins and minerals, enzymes, yeasts, etc, required for a healthy honeybee.

Step 1 

For 1:2 add 6 cups of white sugar to 1 gallon of water. For 1:1 add 11 cups of sugar to a gallon of water. Eyeballing it 1:1 is filling a glass (plastic emits a gas when hot) container about ¾ full of sugar. Put in enough hot water, not quite boiling, to dissolve the sugar (about ½ of a tea kettle). Stir often to dissolve the crystals. I do not use organic sugar as there are particles other than sugar that the honeybee cannot digestible. When the solution has cooled to about 96°, if feels slightly cool to your touch, go to step 2.

Step 2

Add ½ – 1 pint of honey, preferably chemical free and no essential oils.

Step 3

Add 1Tbsp of ACV to the full glass gallon container. Stir. Feed your bees.

I feed the honey formula to my bees once in the spring and 1-2 times in August. Roundup is sprayed in the spring to ready the ground for planting and is sprayed again a few weeks later after the corn is up about a foot. It is sprayed again in August, during the dearth. It is sometimes sprayed again, along with neonicotinoids. Neonics are used to ripen crops.

The premise of BEe Perspective Beekeeping is not putting through the roof what bees do not take through the entrance. I do not consider using the probiotics as “treating” the bees. I consider it as replacing the vital bacteria people kill when using herbicides. My 2 formulas and sometimes extra pollen are the only things I put in the hive.

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[i] http://articles.mercola.com/sites/articles/archive/2003/10/18/bacteria-gut.aspx

[ii] https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4579437/

[iii] https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3989306/

[iv] http://beeaware.org.au/archive-pest/european-foulbrood/#ad-image-0

[v] https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4511666/

ivihttp://journals.plos.org/plosone/article?id=10.1371%2Fjournal.pone.0072106 Microbial Gut Diversity of Africanized and European Honey Bee Larval Instars, Svjetlana Vojvodic , Sandra M. Rehan, Kirk E. Anderson ,  Published: August 21, 2013.

[vii] http://digitalcommons.unl.edu/cgi/viewcontent.cgi?article=1028&context=foodsciefacpub , pH Homeostasis in Lactic Acid Bacteria, 9-12-1993, Robert W. Hutkins University of Nebraska, rhutkins1@unl.edu

[viii]http://www.coloss.org/beebook/I/gut-symbionts/4. Culture conditions for the dominant members of the bee gut microbiota

[ix] http://journals.plos.org/plosone/article?id=info:doi/10.1371/journal.pone.0072106  Microbial Gut Diversity of Africanized and European Honey Bee Larval Instars, Svjetlana Vojvodic, Sandra M. Rehan, Kirk E. Anderson, August 21, 2013.

[x] Lactobacillus sp. as a potential probiotic for the prevention of Paenibacillus larvae infection in honey bees, Dagmar Mudroňová, Juraj Toporčák, Radomíra Nemcová, Soňa Gancarčíková, Vanda Hajdučková, and Katarína Rumanovská. University of Veterinary Medicine, Komenskeho 73, 041 81, Košice, Slovak Republic. 27 September 2011.

[xi] http://aem.asm.org/content/74/24/7750.full.pdf+html APPLIED AND ENVIRONMENTAL MICROBIOLOGY, Dec. 2008, p. 7750–7758 Vol. 74, No. 240099-2240/08/$08.000 doi:10.1128/AEM.00982-08, 2008, American Society for Microbiology. Population Heterogeneity of Lactobacillus plantarum WCFS1 Microcolonies in Response to and Recovery from Acid Stress, Colin J. Ingham, Marke Beerthuyzen, and Johan van Hylckama Vlieg

[xii] http://www.lamission.edu/lifesciences/lecturenote/mic20/Chap06Growth.pdf.  Physical requirements for organism growth.

[xiii]https://www.researchgate.net/publication/8431169_Bacterial_Probiotics_Induce_an_Immune_Response_in_the_Honey_Bee_Hymenoptera_Apidae – Bacterial Probiotics Induce an Immune Response in the Honey Bee (Hymenoptera: Apidae) Jay D. Evans and Dawn L. Lopez

[xiv] http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4700093/. Are commercial probiotics and prebiotics effective in the treatment and prevention of honeybee nosemosis C? Aneta A. Ptaszyńska, Grzegorz Borsuk, Agnieszka Zdybicka-Barabas, Małgorzata Cytryńska, and Wanda Małek, 2015 Oct 6. doi:  10.1007/s00436-015-4761-z, PMCID: PMC4700093

[xv] http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4700093/. Are commercial probiotics and prebiotics effective in the treatment and prevention of honeybee nosemosis C? Aneta A. Ptaszyńska, Grzegorz Borsuk, Agnieszka Zdybicka-Barabas, Małgorzata Cytryńska, and Wanda Małek, 2015 Oct 6. doi:  10.1007/s00436-015-4761-z, PMCID: PMC4700093

[xvi] http://www.janethull.com/newsletter/0704/chlorine-in-your-pool-and-diet-cola.php

[xvii] Hydrion Ph paper (93) with Dispenser and Color Chart – Full range Insta Chek ph- 0-13