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Technical bulletin N°7Technical bulletin

Reminder : definitions and objectives

Barrier flora used in breeding are also called competition flora, competitive exclusion flora or probiotics. Many products are present on the market but choosing the good barrier flora is not easy for the following reasons:

1. The regulatory framework is rather loose depending on the country

2. Product labels are not always very informative

3. The organic label guarantees a good safety and a usage authorization in Organic farms , but gives no guarantee of effectiveness

4. The mode of production and quality control are not subject to specific standards

5. The new generations of barrier flora, compared with the «old» probiotics, base their effectiveness on specific criteria often unknown to users.

The use of barrier flora serves several purposes:

• Occupy colonization sites of pathogenic bacteria to block their implementation, especially in the animal environment : buildings, litter, equipment before the arrival of animals;

• Implement natural digestive bacteria in the intestine of monogastric animals by supporting the functioning of the digestive ecosystem and reducing the growth of pathogenic bacteria through competitive effect;

• Act by preventing harmful bacterial infections and reduce antibiotic use;

• Improve performance through better nutrient absorption;

• Improve the comfort of animals and breeders by reducing anaerobic fermentation which produces high annoyance gases (ammonia , methane , hydrogen sulfide ... ) for the benefit of the favorable aerobic fermentation to maintain good litter ;

• Producing quality compost from litter seeded with composting flora.

The fight against antibiotic-resistant bacteria

This is a major concern for hygienists both in veterinary and human health and has its origin in the expansion of antibiotic usage.All European countries have developed plans to decrease the use of antibiotics; in France, the Eco Antibio 2012 - 2017 plan provides for a 25% decrease in consumption of antibiotics through better application of the treatments, but also by the development of alternatives : barrier floras are included.Resistant bacteria are prevailing as a result of two «survival» mechanisms:1. The selection, after antibiotic treatment, of pre-existing resistant bacteria;

2. The transfer from one bacterium to another, including between different species of resistance plasmids.

If the second mechanism is more difficult to control, however the first one is largely inhibited by reducing the use of antibiotics.Various factors promoting the development of antibiotic-resistant bacteria are added to this phenomenon: the antibiotic use not only as curative but also as preventive mean, the « wear » of older drugs, veterinary treatment restriction to few molecules, and the fact that pharmaceutical research does not discover new molecules. Moreover, even if a new antibiotic would be identified, it would likely remain reserved for human medicine.We are condemned to find alternative solutions

The criteria for choosing a good barrier flora

The safetyAll flora manufacturers typically include bacterial strains referenced as apathogenic by EFSA ( European Food Safety Authority ) and the Institut Pasteur through the CNCM (Collection Nationale de Cultures de Microorganismes) : most often it refers to lactic flora or Bacillus sp. strains. There is therefore no risk of toxicity even at high titers, even less at dosages recommended by the manufacturers; the main limitation is economic.The substrates are culture media under liquid or dried form without toxicity..

The titerThe higher the titer , the stronger the barrier effect; for large farms , it is necessary to provide high concentrations of each strain of bacteria: 1000 bacteria per subject is equivalent to 1 billion per liter of concentrate at the rate of 100 ml to be diluted for 10 000 animals.

Activity spectrumIt is necessary to aim at a broad spectrum of activity to obtain an effective parade in contaminations by numerous species of pathogenic bacteria. Furthermore, several strains of healthy and well chosen bacteria are going to act in synergy between themselves as Bacillus with the lactic flora. Wide spectrum also means specificity of action of a particular flora against a specific pathogenic bacterium. It is the case of the lactic bacteria against Salmonellas.

Target speciesThe selected bacteria are aerobic (need for oxygen for their multiplication). The aerobic fermentation which ensues from it, is a natural process of the digestion of organic matters both in the composting of fertilizers as in the digestive tract, but beware, only in monogastric like poultry , pigs , calves (monogastric yet !). In ruminants (polygastric), digestion involves an anaerobic fermentation which can not be controlled by aerobic barrier flora.

Application programs and their costsWe must compare the programs offered by manufacturers. A continuous application in a livestock- at an effective rate will cost more than spot applications. The cost of a single local nebulization is not comparable to a comprehensive treatment of the environment and of the animals by ingestion. As efficiency depends on the pressure that will be exerted in the farm by the barrier flora, there is no miracle : the more you put, the better the result.

Selection criteria of barrier flora for breeding

the prophylaxis

What to trust to assess the quality of a barrier flora ?Unfortunately, the law is not very precise to give guarantees in particular through product labeling. The user must therefore ask his supplier to form its own opinion.

A quality label ?• The Organic Certification: it is before any, a guarantee of safety and

conformity of the constituents of the products according to the criteria of use in Organic Breeding; on the other hand, this label implies no guarantee in terms of efficiency.

• The French law: it does not require manufacturers to register flora as food additives, especially when they are distributed in drinking water. They are more considered as nutritional supplements over the counter, so no specific standards are imposed

• The European Legislation: applied in the drinking water as well as in the feed, flora are considered as feed additives under the European classification and then submitted to registration the more demanding that the product contains more bacterial strains. That is to say that a cocktail including a large number of bacterial strains, so a priori better, is virtually impossible to register.

• However we can ask whether the manufacturing site meets quality standards

like the Good Manufacturing Practices (GMP), or whether it is approved by the local Authorities.

Production by extraction versus individual strain multiplica-tion

• By extraction : as far as a multi-strains cocktail is concerned, it is common that all strains are multiplied together in the same culture medium. A competition then comes into play which does not guarantee a good title for every bacterium. In addition, a slight variation of the culture conditions affects the composition of the final product, resulting in a potentially variable quality from a production batch to another.

• Individual productions : conversely, this production process where each strain is multiplied individually on its own medium and then mixed with the other strains, guarantees high, homogeneous and controllable individual titers.

Dry or liquid substrateThis choice is important in terms of fragility of the bacteria often susceptible to heat treatments or desiccation. The liquid forms are preferred to achieve and maintain high titers..

Special case of lactic floraThey play a important role in the acidification of the intestine and by secreting prebiotics, with as a consequence, a predominant action anti-Salmonella and against other harmful enteric bacteria.By cons, they are not necessary for composting, Bacillus are able by themselves to sanitize litter or manures in less than 2 months.

Absence of sporulationThis factor leads to fragility, because the vegetative forms are much less resistant than Bacillus spores for example. Lactic acid bacteria are then particularly sensitive to heat and desiccation. It is therefore difficult to obtain high titers in preparations in powder form.

StorageLiquid forms therefore have an advantage, but with the constraint of a refrigeration conservation and a 6 to 18 months shelf life.

We would have a similar result in the presence of E. coli or Campylobacter sp. that do not grow in a medium acidified by lactic flora. The presence of this flora at the right titer is very effective in laboratory tests.

In vitro results of the action of the lactic flora against SalmonellaBacterial competition in the lab

Flora applied in the environmentA good barrier flora is able to colonize inert surfaces and maintain it for three months according to the tests of samples from swabs on surfaces and equipment. It thus establishes a barrier against external contaminants: it is the law of the first occupant.

« The biological disinfection? »Cleaning and meticulous disinfection of a building cannot eliminate 100% of bacteria. Some remaining ones are sufficient to re-contaminate livestock. The barrier flora can occupy «the microbe nests» as in crevices of porous materials such as wood, oxidized metals, insulation plates, chipped equipment, and gaskets… Their presence will temporarily block the intrusion of pathogenic bacteria. Spectacular results were recorded in pigeon flocks in permanent breeding with wooden fittings and nests constituted of cardboard egg trays.In chicken farms in the United States, successive batches are bred on the same litter (Principle of the permanent litter) with very good results: one wonders if maintaining a natural physiological flora from one flock to the next

one, is not favorable to a natural resistance of the livestock facing potential contaminations. Chemical disinfection would therefore intervene when a pathogen would have superseded the natural flora.What to treat in the environment and how?

• What ? All internal surfaces of a building, equipment, litter.• How? by watering or spraying.

How often ?• Once per broiler flock.• Every 2 months for longer production cycles as on a laying farm.• Take advantage of Salmonella mandatory controls to detect the presence of

lactic flora on building walls and adapt the pace of implementation.

Salmonella control Competitor flora + Salmonella

Bacillus flora + Salmonella Lactic flora + Salmonella

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Competitor flora + Salmonella

Lactic flora + Salmonella

Flora distributed to the animals

You have to first choose between a distribution in the food or in the drinking water and according to objective, consider the following:

• «more is better» ; the limitation is purely economic• Carefully choose the product and the program - Follow the manufacturer’s and the veterinary’s instructions - Be conscious that we are in a framework of prevention, not of a curative treatment - Depending on the potential threat or history of livestock, focus on intensive or regular programs or a mix of both• Cautions - Stop any antibiotic treatment - Stop disinfectants et acidifiers• Expected effects of the distribution of a barrier flora: - Seeding or strengthening of good intestinal flora - Acidification of the gut by lactic flora - Bacillus - Lactic Bacteria Synergy - Fight against enteric pathogenic bacteria (even

Composting

Indirectly, composting at the farm, by the use of Micro-Organisms (MO) has a favorable effect on the environment, livestock performance and profitability of the operation by a valuation of a sub product : the litter.

Effect on the environmentThe temperature rise to 65 ° C for 15 days progressively decreasing in 42 days allows sanitizing the manure avoiding to contaminate the environment during transport and application on crops. Antibiotic-resistant bacteria have been found in vegetables grown on land fertilized by non-composted manure.

Composting method by MO• This method is officially recognized by official bodies in France. Check

with your local Authorities.• This is by far, the most economical and environmentally friendly method

compared to those of the turning the piles and forced aeration.• It involves aerobic fermentation (low odor, low product gas), as distinct

from the anaerobic fermentation (biogas).• The only constraints are related to the composition of the incoming

manure (<60% humidity, C / N = Carbon / Nitrogen, about 12).

Selection criteria of the product and compliance with the method

• The presence of bacteria (Bacillus sp) is necessary for good activity; Warning! Some compost activators have only bacteria nutrients without the presence of bacteria themselves.

• The manufacturer must ensure high concentration of bacteria.• The compost enhancer is diluted and distributed by spraying the manure;

homogenizing by passing through a mixing machine is very favorable.• To ensure good oxygenation of windrows, limit their height up to 3

meters; digestion of the organic material provides then naturally the flow of air in the windrow, which explains why it does not require rollover.

It’s necessary to compost its litter !

• There is no operational size requirement;• It sanitizes your farm, but also secures the transport and the application

on crops in terms of health protection;• Apply a healthy fertilizer is reaping healthy vegetables;• Just 42 days to get a sanitized compost, 84 days for a mature compost;• Investment is low and the compost is a real added value.

Finally, use a new generation of barrier flora is:

• Being disengaged from the systematic application of antibiotics and its consequences

• Getting into an organic and / or rational approach to breeding in small or large scale

• Anticipate a voluntary approach which will be imposed at a more or less long term by legislation and consumer pressure

The criteria for product selection and application programs are not complicated but we must require some guarantees of composition and concentration on the part of the supplier, to implement the right program and achieve the best results.

Range Filaflor Products

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NAMES PACKAGINGS DOSAGE METHOD OF USE

FILACTIS LIQUID

500 ml graduated bottles

Poultry : Dose for 20 000 chickens (or 10 000 turkeys, 10 000 laying hens, 10 000 ducks or 10 000 chicks). · In the starting up or in the beginning of production: dose of 100 ml distri-buted on the first 5 days. · Then 100 ml, once a week up to the slaughter or the end of production. (Possibility of making cures of 5 days, renewable according to needs, during risk period). Pigs: Dose: 10 ml / ton of live weight every day during 5 days.

Drinking water

100 ml graduated bottles

FILAFILM LIQUID

1 liter graduated bottles

Spraying of surfaces : 1 dose of 100 ml of FILAFILM LI-QUID for 1000 m2 of total surface or 4 000 birds in cagesSpraying ot the eggs: 1 ml de solution diluée pour 500 oeufs

Spraying500 ml graduated bottles

100 ml graduated bottles

HYGIENE PRODUCTS

DESIGNATIONS PRESENTA-TIONS

DOSAGE APPLICATION

FILAFLOR COMPOST

Tin of 10 liters 1 liter for 10 metric tons of manure with a minimum efficient dose of 5 liters

Incorporation Tin of 5 liters

FILAGREEN Tin of 1 liter 100 ml for 10 liters of material to be composted Incorporation

ENVIRONMENT PRODUCTS

Products approved for use in organic agriculture in compliance with the (CE) N°834/2007 and NOP regulation.