September | October 2010

Feature title: Ecobiol Aqua the effective single strain probiotic

The International magazine for the aquaculture feed industry

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Probiotics are viable cultures of microorganisms which, when added to the animal feed, can reach and colonise

the animal’s digestive system, benefi-cially improving the properties of the indigenous flora and interacting with the pathogenic bacteria.

Probiotics in aquaculture can work on two completely different situations: on one hand, they can control pathogenic bacteria population in animals’ intestines (competitive exclusion, bacteriocines), as well as help the animal digest the feed and maintain good health; they are considered as a good option to antibiotic growth promoters.

On the other hand, the addition of live bacteria to the ponds where the animals are farmed may modify the bacterial com-position of the water and sediment.

In aquaculture, it is easier for the opportunistic pathogenic bacteria to colonise the gastrointestinal tract of the animals because they are supported by the environment where the animals are farmed.

For example, Vibrio bacteria are nor-mally found attached to algae and forms part of the normal flora of fish and shrimps as well as zooplankton.

In aquaculture ponds, where animal and

algal population densities are very high, Vibrio numbers are much higher compared to the normal open water situation. In these cases it a defence against over-exposure to Vibrio is necessary (whose pathogenicity may have been increased by the abusive use of antibiotic growth promoters).

Survive in adverse conditions

Ecobiol Aqua is a probiotic developed by Norel SA that is made up exclusively of spores of Bacillus amyloliquefaciens.

The spore is a struc-ture that is adopted by the bacteria to survive in adverse conditions.

During the produc-tion process in León (Spain), the vegetative bacteria of B. amyloliquefaciens are forced into the spore form. This confers the bac-teria a very stable presentation for storage and feed processing, without affecting its capacity to acquire the vegetative form once the environment is favourable for its development (that is, inside the animals, where there are nutrients, adequate tem-perature and moisture).

Different trials have demonstrated that

the spores of the B. amyloliquefaciens can endure pelleting processes of up to 90ºC (see Table 1). If the feed processing reaches higher temperatures than this, it is recommended to apply the product post pelleting (mixed with fish oil, for example).

The effect of Ecobiol Aqua is due to several reasons:

Ecobiol Aqua improves nutrient digest-ibility of the diet because of the produc-tion of enzymes once inside the animal

The name of the bacteria included in Ecobiol Aqua, B. amy-loliquefaciens, was given after observing the way starch was hydrolysed in the presence of these bacteria. But B. amyloliq-uefaciens produces not only amylases: it also produces proteases and lipases that help the ani-mal digesting the feed.

In a study performed with Ecobiol Aqua in shrimp feed (presented at the ASAIM SE Aquaculture Conference 2010), it was observed that two hours after the administration of feed, the animals that were receiving Ecobiol in the feed - at a ratio of 106 CFU/gram of feed - had higher enzymatic activity than the control animals (no probiotic).

The amylase activity was 315.4 units

Ecobiol Aquathe effective single strain probiotic

Table 1: - B. amyloliquefaciens counts (CFU/g of feed) before and after pelleting (90ºC)

Before pellet After pellet

2.60 x 106 2.25 x 106

1.68 x 106 1.50 x 106

1.27 x 106 1.16 x 106

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F: Probiotic

IAF10-05.indd 18 07/09/2010 14:55

in the probiotic group, 5% higher then the 302 units in the control group. The protease activity was 7.47 units in the probiotic group, almost two times the four units measured in the control group. The differences in the lipase activity were not so high, only 2.5% higher, 40.32 units in the probiotic group and 39.1 in the control group.

From this we deduce that this enzyme production improves not only protein, starch and fat digestibility but also the energy digestibility, as these nutrients are essential for the energy of the diet calculation.

Ecobiol Aqua controls intestinal pH by producing lactic acid

Bacteria ferment carbohydrates to obtain energy following different biochemi-cal pathways; in the case of B. amyloliquefa-ciens, carbohydrates are normally degraded following the lactic acid route.

It has been observed that for every gram of sugar present in the media, B. amylolique-faciens is able to produce 0.9g of lactic acid. If there are enough carbohydrates in the environment, 1012 CFU of B. amylolique-

faciens (nor-mal content of bacteria in one tonne of feed) pro-duce 1.8kg of lactic acid. When Ecobiol pro-duces the lactic acid in the fish’s gut, it controls pathogen ic bacteria and is used as substrate for growth by beneficial bacteria (lactobacilli). Several trials have demon-strated the beneficial effect that lactic acid has on fish growth.

Ecobiol Aqua actively balance intestinal microbiota

Once the spore is transformed into the active form of the bacteria, the B. amyloliquefaciens produces Barnase to avoid competition for the loci. Barnase is a ribonuclease that, without its inhibitor,

Barstar (produced by the same bacteria but not ejected to the media), acts as a natural antibiotic.

Barstar is active at all pHs and is effective inhibiting the transcription of the genetic material into protein in all kinds of bacteria, Gram negative (E. coli, Vibrio) and Gram positive (clostridia).

The inhibition of growth of those bac-teria has been demonstrated in different studies evaluating the bactericide capacity

Table 2:- Effect of the addition of a B. amyloliquefaciens probiotic on mortality

Treatment Ponds Surface (has) Initial animals Mortality (%)

Control 3 24.5 1960000 39.34

Ecobiol Aqua 3 24.5 1960000 33.50

Table 3:- Effect of the addition of a B. amyloliquefaciens probiotic on production

Treatment Ponds Surface (ha)

Initial animals

Final Biomass

(Ton)

Final Body Weight (g)

Control 3 24.5 1960000 19.6 16.90

Ecobiol Aqua 3 24.5 1960000 23.6 18.16

18 | InternatIonal AquAFeed | September-october 2010 September-october 2010 | InternatIonal AquAFeed | 19

F: Probiotic

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MARKET TRENDSCurrent and Future Market Trends in Aquaculture

FEED INGREDIENTSIngredient Trends and the Effects on the Extrusion Process; Utilization of Algae in

Aquafeed and Nutrition; Raw Material Bottlenecks in Formulation

PRACTICAL CONSIDERATIONS IN FEED FORMULATIONFeed Formulation and Nutrition; Artemia, Enrichments and Other Feed Additives for

Hatcheries; Larval Nutrition and Starter Feeds; Shrimp Feed: Formulation and Stability; Rapid Analytical Methods for Ingredient Testing

FEED MANUFACTURING TECHNOLOGYGrinding for Aquatic Feed; Feed Preparation Technology; Feed Coating, Absorption

Coating by Vacuum and Micro Encapsulation; Feed Pelletization; Proper Dosifi cation & Mixing of Macro-Minor Ingredients and Liquids for Aquafeed,

AQUATIC FEED EXTRUSIONFundamentals of Introduction to Extrusion Technology; Critical Issues in Extruder

Process Control; Aquatic Feed Extrusion: Applications of Twin Screw Extruders; New Extrusion Technology with Online Control of Product Density and Specifi c Mechanical

Energy Input; High Fat Extrusion for Aquafeed; Micro Aquatic Feeds; Drying of Aquafeed

PLANT DESIGNCritical Parameters and Plant Design for Aquafeed; Environmental Issues: Noise and

Odor Control; Post Processing and Handling of Quality Aquafeed

QUALITY ASSURANCEAquafeed Quality: Lipid Oxidation and Palatability; Chemical and Microbiological

Aspects; Mycotoxins Management in Aquafeed; Verifying End Product Quality

CONTACTS

Dr. Ignace DebruyneID&A Ignace Debruyne & AssociatesTelephone: +32 (0)51 31 12 74Fax: +32 (0)51 31 56 75Emai: aquafeed@scarlet.be

Dr. Sefa KoseogluFiltration and Membrane World LLCTelephone: +1 979 764 8360Fax: +1 979 694 7031sefa.koseoglu@membraneworld.com

10TH PRACTICAL SHORT COURSE:Aquafeed Platform - Aquaculture Feed Extrusion,

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4 | InternatIonal AquAFeed | September-october 2010 September-october 2010 | InternatIonal AquAFeed | 5

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so the quality of the water quality becomes increas-ingly important for animal survival.

Mortality is also reduced as the probiotic improves animal intestinal health and hence, general health status; it being more difficult for pathogens (even virus) to successfully infect an individual.

In a field trial run with a total of six ponds, 49ha and 3,920,000 shrimp, the mortality was reduced more than 5%.

Ecobiol Aqua improves feed conversion ratio and growth (animal size)

Due to all the afore-mentioned benefits: enzyme production, lactic acid excretion to the lumen, intestinal micro-biota balance, better ani-mal health, etc, the animals are able to perform better from a zootechnical point of view.

Ecobiol Aqua is effec-tive in both young and

adult, productive animals.The effect on young animals has been

demonstrated in different trials (see Figure 2). It helps the earlier uptake of feed and avoids health problems.

With adult animals, when the density in the ponds increases and the water quality worsens, the effect of the probiotic is even more effective (see Table 3).

ConclusionThe usual probiotic products used in

aquaculture are composed of different strains of different bacteria that do not have well defined effects. Ecobiol Aqua is a single strain probiotic effective at many different levels that ensures safe and economically profitable production.

of the supernate of B. amyloliquefaciens. The supernate of the fermentation of B. amyloliquefaciens is placed in six mm wells, and the halo around that well where the dif-ferent shed bacteria are not able to grow is measured: the bigger the halo, the stronger the inhibition.

The combination of the Barnase produc-tion, that inhibits the growth of pathogenic bacteria populations, with the lactic acid production, that improves the lactic flora population, ends up in a well-balanced, steady and healthy intestinal microbiota in the fish, which can be observed as better health and more resistance to illnesses in the animals.

Ecobiol Aqua reduces mortality in the ponds

As mentioned before, the use of the probiotic Ecobiol Aqua improves nutrient digestibility. This fact has a direct effect on the water quality, there is less organic matter in the water, and whatever organic matter is in there, can be degraded by the bacteria included in the probiotic. In aquaculture the animal density is much higher than in natural open water situations

Figure 1: Inhibition halos of Vibrio parahaemolyticus CECT 511 around the wells of Ecobiol Aqua supernate

Figure 2: Performance parameters in Litopaneus vannamei juveniles fed a control diet, or a diet including B. amyloliquefaciens probiotic

More inforMation:Juan José Mallo, Ingeniero AgrónomoTechnical Director Norel S.A.P.I. Can Coll, c/ Indstria, 1(08185) Lli de Vall, Barcelona SpainWebsite: www.norelynature.com

"Ecobiol Aqua is a single

strain probiotic effective at many

different levels that ensures safe and economically

profitable production"

20 | InternatIonal AquAFeed | September-october 2010

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VOLUME 13 I S SUE 5 2 010

Influence of natural and artificial binders

- in feeds for Litopenaeus vannamei on digestibility & growth

Ecobiol Aqua - the effective single strain probiotic

A general overview aquaculture in the EU

Abalone feed development in South Africa

IAF10-05.indd 1 07/09/2010 14:54

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