Lactobacillus

acidophilus

D2/CSL

(CECT 4529)

HISTORY

SAFETY

EFFICACY

1985 to 2018

4 SYNERGETIC COMPANIES

Today, four different companies work in synergy within a single business system, validly maintaining their autonomy, characteristics and production specificity.

AGROZOOTECHNICAL CULTURES

AGRO ZOOTECHNICAL

POULTRY

SILAGE

MATERIALS:

- FOODS

- FEEDS

- EXCRETA...

Isolation of LAB strains

Characterization

SelectionProduction

Strain collection

Pilot plant

HOW CSL WORKS

Lactobacillus

acidophilus

D2/CSL

CECT 4529

HISTORY

1985 – CSL isolates 19 strains of faecal lactobacilli from a «rural» adult chicken

1985 – Choice of L. acidophilus D2/CSL from the 19 LAB strains

The choice of the strain L. acidophilus D2/CSL over the other 18 isolated strains was due to:

• better bile-resistance

• better adhesion to the gut epithelium

• better resistance to the jonophorous antibiotic

• better performance in the production plant (recovery of UFC after the freeze-drying process)

Lactobacillus

acidophilus

D2/CSL

CECT 4529

SAFETY

The species L. acidophilus is considered by EFSA to be suitable for the Qualified Presumption of Safety (QPS) approach to safety assessment (EFSA, 2007; EFSA BIOHAZ Panel, 2013).

This approach requires:

- THE IDENTITY OF THE STRAIN to be conclusively established and

- evidence that it does not show RESISTANCE TO ANTIBIOTICS of human and veterinary importance.

SAFETY OF L. acidophilus (FEEDAP, 2014)

NOT TRASMISSIBLE

2012 – Results of the last A-R test (FEEDAP, 2012)

Lactobacillus

acidophilus

D2/CSL

CECT 4529

EFFICACY

1993 – Efficacy trial - D2/CSL – laying hens

- ISA WARREN HENS – KEPT IN CAGE- TRIAL 1 = 7150 HENS - 3 GROUPS = 2 TR + 1 CO- TRIAL 2 = 672 HENS - 8 REPETITIONS = 4 TR + 4 CO - PROBIOTIC IN DRINKING WATER

1994 – Efficacy trial – D2/CSL – broilers

- SHAVER REDBRO BROILERS – KEPT ON LITTER- TOT. 216 CHICKS - 12 GROUPS = 6 TR + 6 CO- PROBIOTIC IN DRINKING WATER

1995 – Efficacy trial - D2/CSL – broilers

- SHAVER REDBRO BROILERS – KEPT IN CAGE = 3 CHICKS/CAGE- TOT. 54 CHICKS - 18 CAGES = 9 CAGES TR (27 CHICKS ) + 9 CAGES CO (27 CHICKS)- PROBIOTIC IN DRINKING WATER

2007 – No. 3 «in field» efficacy trials - broilers

- ROSS 508 BROILES – KEPT ON LITTER - 90.000 CHICKS – 3 SEASONS – 30.000 CHICKS/SEASON- 6 POULTRY HOUSES – 3 TR + 3 CO (15.000 CHICKS/SHELTER)- PROBIOTIC IN DRINKING WATER

2008 n. 1 – Efficacy trial – laying hens

- HI-LINE BROWN HENS – KEPT IN CAGE- 160 HENS - 8 GROUPS = 4 TR + 4 CO - PROBIOTIC IN FEED

2008 n.2 – Second efficacy trial – laying hens

- HI-LINE W 36 HENS – KEPT IN (SINGLE) CAGE- 120 HENS – 3 GROUPS = CO (D2) ; TR1 (D2 + SMP

3%); TR2 (D2 + SMP 4%) - PROBIOTIC IN FEED

2008 n. 3 – Study of an analysis method for feed

Ital.J.Anim.Sci. vol. 7, 405-409, 2008

2012 – Efficacy trial of «MIX AVI» - laying hens

- HI-LINE BROWN HENS – KEPT ON LITTER- 2 POULTRY FARMS – 480 HENS IN TOTAL- 2 GROUPS PER FARM = CO; TR (D2 + 14D)- 3 REPLICATES PER GROUP- PROBIOTIC VIA THE DRINKING WATER

2014 – Efficacy trial- laying hens

- HI-LINE W-36 HENS – KEPT IN CAGE- 2400 LAYERS- 3 GROUPS = CO (D2) ; TR1 (D2 + SMP 3%); TR2 (D2 + SMP 4%) - PROBIOTIC IN FEED

2015 – Authorization for D2/CSL (CECT 4529) in the EU: laying hens (feed + water)

2017 – Authorization for D2/CSL (CECT 4529) in the EU: chickens for fattening

COMMISSION IMPLEMENTING REGULATION (EU) 2017/2275ANNEX

Identifi-

cation

number of

the additive

Name of the

holder of

authorisation

Additive Composition, chemical formula,

description, analytical method

Species or

category of

animal

Maximum

age

Minimum

content

Maximum

content

Other provisions

End of

period of

authori-

sation

CFU /kg of complete

feedingstuff with a moisture

content of 12%

Category of zootechnical additives. Functional group: gut flora stabilisers

4b1715 Centro

Sperimentale del latte

Lactobacillus

acidophilus CECT 4529

Additive composition:

Preparation of Lactobacillus acidophilus

CECT 4529 containing a minimum of: 5x1010 CFU/g of additive (solid form).

---------------------------------

Characterisation of the active

substance:

Viable cells of Lactobacillus acidophilus

CECT 4529.

-----------------------------------

Analytical method1

Enumeration: spread plate method using

MRS agar (EN 15787).

Identification: Pulsed Field Gel

Electrophoresis (PFGE).

Chickens

for fattening

- 1 x 109 - 1. In the directions for

use of the additive

and premixture, the

storage conditions

and stability to heat

treatment shall be indicated.

2. For users of the

additive and

premixtures, feed

business operators

shall establish

operational

procedures and

organisational

measures to address

potential risks

resulting from their

use. Where those

risks cannot be

eliminated or

reduced to a

minimum by such

procedures and

measures, the

additive and

premixtures shall be

used with personal

protective

equipment,

[10 years

from the

date of entry

into force of

this Regulation

To be

completed

by the

Service

responsible

for the

publication]

1 Details of the analytical methods are available at the following address of the European Union Reference Laboratory for Feed Additives: https://ec.europa.eu/jrc/en/eurl/feed-additives/evaluation-reports

2015-2018 The 5 more recent experimental trials

1. UniPg-IZSUM & UniTe: D2/CSL in organic layers

2. UniPg & IZSUM: D2/CSL in organic layers

3. UniBo: D2/CSL on broiler performance

4. UniBo: D2/CSL on intestinal chicken microbioma

5. UniPg & IZSUM: D2/CSL in rural broilers

2015 – Efficacy trial UniPg-IZSUM-UniTe – organic hens

2016 – Efficacy trial UniPg-IZSUM – organic hens

2016 – Efficacy trial UniBo – intestinal microbioma and metaboloma of broilers

2017 – Efficacy trial UniBo – D2/CSL effects on performance and microbioma of broilers

2018 – Efficacy trial IZSUM-UNIPG – D2/CSL effects on rurally reared broilers

1985-2018: summary of the efficacy resultsLaying hens

The better “GUT HEALTH” (or EUBIOSIS) due to L. acidophilusD2/CSL has led to significant improvements in terms of:

1) Intestinal microbiota composition

2) FCR (kg feed/kg eggs)

3) Egg production

4) Egg quality (ESG, shell thickness, Haugh Units)

5) Saleable eggs (less wastage: dirty eggs, cracked eggs, etc.

6) Environment: reduction of volatile N compounds and faecalmoisture within the poultry house.

1985-2018: summary of the efficacy resultsBroilers

The better “GUT HEALTH” (or EUBIOSIS) due to L. acidophilusD2/CSL has led to significant improvements in terms of:

1) Gut microbiota modifications

2) Gut morphology (villus height)

3) Morbidity and pasty vents

4) Weight increment

5) FCR (Kg feed/kg weight increment)

6) Environment: reduction of volatile N compounds and faecalmoisture within the poultry house.

SUMMARY OF THE ACTIVITIES OF L. ACIDOPHILUS D2/CSL

L. ACIDOPHILUS D2/CSL

GASTRO-INTESTINAL

MICROBIOTA

PRODUCES BY FERMENTATION

REBALANCES THROUGH

ACIDIFICATION

REBALANCES BY

COMPETITION WITH

PATHOGENS

MORE GUT HEALTH AND ANIMAL PRODUCTIONS

LACTIC

ACID

GALT

DEVELOPS

AND STIMULATES

Commercial chickens are subject to INTESTINAL DYSBIOSIS.

The outcome can be a TRUE PATHOLOGY or, more frequently, a SUB-CLINICAL “MALAISE” that worsens the flock PERFORMANCE.

The INTESTINAL DYSBIOSIS:

produces INFLAMMATION and alters the DIGESTIVE FUNCTIONS;

damages the EGG PRODUCTION in quanti-qualitative terms (dirtyand thin egg shells, liquid albumen, etc.);

damages the BROILER PERFORMANCES and increases BROILERMORBIDITY;

reduces the FEED DIGESTIBILITY and the FAECAL CONSISTENCY

harms the ENVIRONMENT in the shelter (more pathogens, humidity,ammonia, H2S, etc.).

CONCLUDING REMARKS

Each flock is composed by a mixture of “HEALTHY“, “SUB-CLINICALLY ILL“, and “CLINICALLY ILL” birds.

The primary aim of feeding birds with L. acidophilus D2/CSLis to prevent or resolve the gastro-intestinal dysbiosis,namely to protect the “HEALTHY” birds (PROPHYLAXIS)and/or "SUB-CLINICALLY ILL“ birds (METAPHYLAXIS).

That is why L. acidophilus D2/CSL is the “NATURAL WAY” toimprove the productive performance of the so called“HEALTHY” flocks.

CONCLUDING REMARKS

Probiotics and pathogens: the DOSE make the difference!

THE “DOSE” OF D2CSL MUST BE HIGH ENOUGH TO EFFECTIVELY “INFECT” THE HOST.

On the contrary a “cause-effect” relationship is not evident probably because you havegenerated a ASYMPTOMATIC CARRIER!

ASYMPTOMATIC CARRIER(NO EFFECT)

PATHOGENIC STRAIN

LOW (NON INFECTING) DOSE HIGH (INFECTING) DOSE

PROBIOTIC STRAIN

PATHOLOGY

HEALTHINESS

takes care of the gut health

since 1948

THANK YOU FOR YOUR ATTENTION!