JEFO PROTEASE IN AQUACULTURE A UNIQUE SOLUTION

Florence Philippe
PHILIPPE F. Aquaculture product manager, Jefo Europe, 2 rue Claude Chappe, 44481 Carquefou, France

BACKGROUND Study 2:
A 14-week two-treatment growth trial was conducted with 1200 rainbow trout (IBW
Use of protease in aquafeed has been on the rise mainly because their ability to
~60g), distributed into six tanks (200 in each tank). Marine protein and plant protein
reduce the variations in the raw material quality by improving the digestibility of poor
content in control diet were 16% and 25%, and in treatment diet were 9% and 35.8%,
quality protein sources. They provide feed manufacturers much needed room to be
respectively (Fig 3).
competitive without sacrificing the feed quality.
The treatment diet was formulated using Jefo Salmonids Matrix by modifying the
The Jefo protease is a multi-component protease complex with unprecedented
control diet. A slightly higher amount of lysine (1% compared to 0.67%) was added to
versatility on raw material choice.
the treatment diet to balance the amino acid profile.
This paper will discuss two studies conducted in European conditions with Jefo protease
The diets were formulated to be isoproteic (42% CP) and isoenergetic (19 MJ/kg).
(Jefo_PRO).
When analyzed, calcium content was significantly higher (1.8% compared to 1.1%) and
The first, improving digestibility of hydrolyzed feather meal in a a warm-water species, fat and phosphorus in the control diet (24.1% and 1.02%, respectively) were slightly
European sea bass Dicentrarchus labrax . The second trial was assessing growth higher than the treatment diets (22.4% and 0.9%, respectively).
performance of a cold-water species, rainbow trout Oncorhynchus mykis with high
marine and high plant + Jefo protease diets. For the high plant diets, Jefo matrix for Figure 3. Marine proteins (%) in control and test diets
salmonids was used to reformulate the control diet.
18,0
15,0 16.0
Study 1:

% Marine protein
12,0 Control
Feather meals were produced in a commercial rendering plant. Two commercially
available proteases (COM_PRO and Jefo_PRO) were tested against the control. 9,0 Test
9.5
COM_PRO is a commercial protease claiming to contain keratinase. The production 6,0
process was slightly modified to allow enough time for incubation (Fig 1).
3,0
A total of six batches of feather meals (two control and two for each protease) was
produced and subjected to digestibility trial with European sea bass (IBW ~50g) 0,0
following the Guelph method. Fish were acclimated for five days before commencing Marine protein, % feed
the collection of feces. Apparent digestibility of energy and nutrients of the test
ingredients were calculated based on Bureau & Hua (2006).

Figure 1. Schematic diagram of the processing of feather meal between the treatments despite slightly better results in those fed the high marine
protein regular diet (Fig 4).
Slaughter Trommel
(storage) Press
Figure 4.
H2O removal

H2O removal
2,00
Drying:
90 min Bin
Outside feathers
(storage)
1,50
1.60 1.55
Control
1,00
Test
Bin
Dryer Batch Cooker 0.82 0.85
(intermediate storage) 0,50

0,00
Steps:
- Loading + ENZYME (30 min) TGC FCR
- Incubation with enzyme (45 min; 45-50C)-
discount loading time?

Packaging
- Pre-drying (100C; 1 bar; 80 min)
- Pressure/temperature rise (25 min) - less because
CONCLUSION
Storage/Cooling
Big-Bags no need to reach 133C/3bar?
- Sterilization (120-125C; 2 bar; 15-20 min)
- Pressure release (10-15 min)
Hydrolyzed feather meal with Jefo protease can utilize dietary proteins
- Unloading (15 min) significantly better than other products.
Jefo Salmonid Matrix can be a useful tool in salmonids formulation for protein
cost optimization without sacrificing the performance.
Cost savings using Jefo Salmonid matrix in this study was 2% or about 20
USD per metric ton of feed.
feather meal was slightly higher than the other two. However, ADC of crude protein of
JEFO_PRO treated feather meal in European sea bass was significantly higher than Similar savings can be obtained with simple modification of existing formulation
COM_PRO treated feather meal. (i.e., 1-2% change in raw material composition) for any species while not using
matrix.
Figure 2.
treated feather meal in European sea bass References

100% FAO 2015

Bureau and Hua, 2006, Aquaculture, 252:103-105.
90% b
ab a
Control
80%
COM_PRO
70%
JEFO_PRO
60%
50%
Dry Matter Protein Energy Fat
BETTER GROWTH AND INTESTINAL HEALTH OF
RAINBOW TROUT FED DIETS SUPPLEMENTED WITH GRADED
LEVEL OF A PROTEASE
A. Guajarado1, P.C. Villarroel2, R. Gauthier3 and M.A. Kabir Chowdhury3*
1
Universidad de Chile, Chile, 2Alinat Corporation, Chile, 3Jefo Nutrition Inc., Canada

INTRODUCTION Table 1. Growth, production performance and size (m) of intestinal villi of rainbow trout fed a
commercial diets containing graded level (0,175 and 250 ppm) of a protease.

Enzymes allow increasing inclusion of poorly digestible raw materials in the ingredient Thermal-
Initial body Final body Specific
matrix, therefore, reducing the overall formulation cost of a feed. Treatments unit Growth FCR Villi size
weight weight growth rate

In recent years, use of enzymes in aquaculture feed is increasing due to rising cost (g) (g) (SGR) (TGC) (m)
of quality raw materials and volatility in their availability in global market.
Control 390 850a 0.92a 2.52a 1.43b 630a
In this study, graded level (0, 175 and 250 ppm) of a protease enzyme from Jefo Control +
Nutrition Inc., Canada was used in the feed of rainbow trout (Oncorhynchus mykiss). 175 ppm 402 971b 1.05b 2.94b 1.35a 663b
protease
Control +
250 ppm 399 987b 1.07b 3.03b 1.33a 737b
protease
in feed on growth, feed conversion and intestinal health of the test animals.

METHODS Figure 2. Intestinal villi structure and their organization in rainbow trout fed commercial diets with
and without a protease.

> Three commercially extruded rainbow trout diets of 40% CP were formulated CONTROL
to contain 0 (control), 175 and 250 ppm of Jefo protease.

> The experiment was conducted in triplicates in 2-m3 tanks with 40 fish per tank.

> Initial body weights of fish were ranged between 390 and 402 g fish-1, which
were fed the three experimental diets (1.5% BW per day) for 12 months until
they reach the market size of ~1 kg.

> Fishes were sampled every month for growth and adjustment of feeding ration. PROTEASE

> At the end of the experiment, weight gain, growth (SGR & TGC - thermal-unit

> Size and condition of intestinal villi and pyloric caeca of five fish from each tank
were also examined after the experiment.

RESULTS
The growth of fish fed the control diets (without protease) was significantly lower CONCLUSION
than those fed the diets containing protease (Figure 1). Despite slightly better

fish fed the protease diets (Table 1).

of rainbow trout were observed.
The size of intestinal villi (m) also increased linearly with increasing levels of protease
in the diets (Table 1). In addition, the structure, organization and density of villi were
both the protease supplemented diets despite linear increase in villi height suggested
much better in fish fed both the protease diets (irrespective of concentration of the
that most of the dietary proteins were hydrolyzed by protease at the inclusion level
protease) than those fed the control diets (Figure 2).
of 175 ppm.
Figure 1. The incremental growth of rainbow trout fed three commercial diets containing graded
level (0, 175 and 250 ppm) of a protease.
and intestinal health of rainbow trout fed a commercially extruded diet containing
1000
graded levels of the enzyme.
Control 175 ppm 250 ppm
Average fish weight (g)

900
800
700
600
500
400
300
0 1 2 3 4 5 6 7 8 9 10 11
Months Poster 020
EVALUATION OF A PROTECTED B VITAMINS BLEND
VS PROTECTED CHOLINE ALONE ON HEALTH
AND PERFORMANCE OF DAIRY COWS
Leclerc H*, Evans E, Molano R & Lipka A.
hleclerc@jefo.ca

Presented at the XVI Congreso Bienal AMENA, october 2013

INTRODUCTION In this trial, the reduced percentage of tested cows having high blood BHB levels with
The transition period is a critical time period for the dairy cow as she is susceptible to the supplementation of the protected B vitamins blend resulted into improved health,
metabolic events that impact production and reproduction. The period is fraught with milk production and reproductive performance, this is supported by the findings of
many diseases manifested clinically and subclinically. Ospina et al., 2010, showing the negative effects of high blood BHB levels.

OBJECTIVE: Determine if feeding transition cows the protected B

vitamins blend (folic acid, riboflavin and choline) will result in Table 1. Blood BHB results and prevalence of subclinical ketosis.
differences in health status, milk production or reproduction when
compared to protected choline alone. Variable Protected Protected B
Choline1 Vitamins

Number of cows 54 61
MATERIAL AND METHODS
Commercial farm, WA, USA: 1500 dairy cows, free stalls, typical North American total Average blood BHB, mmol/L. 1.524a 0.977b
mixed ration (TMR)
Prevalence of subclinical ketosis, % 31.5a 11.5b
Control: protected choline (Balchem Corporation) 21 days pre to 21 days post
a, b differs P<0.05
calving. 30 % of first calf heifers in both groups
Treatment: protected B vitamins blend (folic acid, riboflavin and choline) (Jefo 1
Pre treatment period only

Nutrition Inc.) from 21 days pre to 21 days post calving.

Design: A three period switchback design study

Figure 1 - Milk yield at the first milk test day.
February April June August 51
50
50.6b
2011 CONTROL 2011 49
48
47
2012 CONTROL 2012 PROTECTED B VITAMINS CONTROL 2012 46
46.4a
45
kg 44
43
42
41
Blood beta hydroxybutyric acid (BHB) measurement: 40
39
38
37
Sub population of multiparous and first calf heifers 36 37.1a
35 35.8a
Every two weeks 34
PROTECTED PROTECTED PROTECTED PROTECTED
Precision Xtra(Abbott Laboratories) meter and blood ketone test strips. CHOLINE B VITAMINS CHOLINE B VITAMINS
Average test day was 7.5 (5-12) days in milk (DIM) First calf heifers Multiparous cows
Blood BHB threshold of 1.4 mmol/L for prevalence of subclinical ketosis. P=0.34 a, b differs P=0.01

No difference between days in milk (ave=24 DIM)

Diseases incidence, milk production at the first milk test day and reproductive
performance were calculated from all the cows calving during each treatment periods
and data were collected from the farm using Dairy Comp 305.

Statistical analysis were conducted using Minitab 16. Differences in disease incidences
and reproductive rates were determined using a Fishers exact test. A two tailed T test
was used to assess treatment effects upon milk production.
RESULTS AND DISCUSSION
Blood BHB levels, the prevalence of subclinical ketosis and milk production are shown
in Table 1 and Figure 1. The cows fed the protected B vitamins blend had less total
clinical illness from 0-30 DIM (30.9% Vs. 24.5% for the control and test group
respectively, P=0.032) with a significant reduction in the incidence of clinical ketosis
(8.3% Vs. 3.0% for the control and test, P=0.001) than the cows supplemented with the
protected choline alone. The incidence of the other diseases (milk fever, displaced
abomasum, mastitis, metritis and retained placenta) was not different between
treatments. A higher percentage of cows bred became pregnant (78.6% Vs. 84.4% for
the control and test group respectively, P=0.054) when supplemented with the
protected B vitamins blend. No significant differences in any of the other reproductive
parameters were observed.
CONCLUSION
DAIRY COWS SUPPLEMENTED WITH THE PROTECTED B VITAMINS
BLEND (FOLIC ACID, RIBOFLAVIN AND CHOLINE) SHOWED A
LOWER BLOOD BHB LEVEL, INDICATION OF A LOWER PREVALENCE
OF SUBCLINICAL KETOSIS, AND A BETTER HEALTH,
REPRODUCTIVE AND MILK PERFORMANCE THAN THE CONTROL
COWS SUPPLEMENTED WITH THE PROTECTED CHOLINE ALONE.
IMPLICATIONS
This trial showed the higher potential of the blend of protected folic acid,riboflavin
and choline compared to protected choline alone, for preventing metabolic
problems, improving health and potentiate productive and reproductive
performance. Dairy producers can consider adding a protected B vitamin blend as a
means of reducing stress during the transition period, this will translate into
economical benefits.
REFERENCES
Ospina PA et al. 2010. J.Dairy Sci. 93:3595-3601
T352 | EFFECT OF RUMEN PROTECTED B
VITAMINS SUPPLEMENTATION DURING THE
RECEIVING PERIOD ON THE PRODUCTIVE
PERFORMANCE OF BEEF CATTLE
H. Leclerc*1, D. A. Espinosa2, E. Evans3, R. Zambrano Gaytan2, J.D. Garza Flores2

1
Jefo Nutrition, St-Hyacinthe, QC, Canada,
2
Rancho El 17, Hermosillo, Sonora, Mexico,
3
Technical Advisory Services, Bowmanville, Ont. Canada
H.LECLERC

INTRODUCTION RESULTS

Because rumen microorganisms are able to synthetize B vitamins, it has long been considered
that it is not necessary to supplement them in the diet of functional ruminants. However,
recent studies have shown that the amounts of B vitamins synthetized in the rumen were not
sufficient to cover the needs of high producing animals (1). Moreover, due to high ruminal
degradation (2), B vitamins need to be protected when supplemented in the diet.
OBJECTIVE
Days to recover shrink were reduced by 1.5 days for the group of cattle fed
the protected B vitamins blend.
No significant difference was observed between the control and treatment group,
for morbidity status (2.16 vs. 1.52%) and mortality rate (0.43 vs.0.87%).
The inclusion of the protected B vitamins blend in the diet significantly improved
total gain by 9% (3.86 kg), average daily gain by 9.5% (0.190 kg) and
feed/gain by 10%; without affecting average daily feed intake (table 2).

To evaluate the performance of finishing beef cattle given a rumen protected B vitamins Table 2: Effect of a dietary supplementation of protected B vitamins blend on performance of beef cattle.
blend during a 21-day feedlot receiving period.
Control Protected B vitamins blend Difference P value
MATERIAL AND METHODS
Number of animals 448 451

Control: Days on feed 21 21

15 No B vitamins supplementation
Treatments:
Initial Weight, Kg
Final Weight, Kg
264.60
305.86
259.26
305.53
5.340
0.330
NS
NS
899 head 2g/head/day of rumen protected B vitamins
(12 loads of cattle) pens/treatment. (folic acid, pyridoxine, pantothenic acid and biotin; Jefo Nutrition, Total Gain, Kg 42.40 46.26 3.860 0.037
St-Hyacinthe, Quebec, Canada).
equally split based on Average Daily Gain, Kg 2.01 2.20 0.190 0.030
weight and shrink.
Feed Intake, Kg 8.53 8.35 0.180 NS
Figure 1: Ingredient composition of diet fed during the trial Table 1: Nutrient composition of the diet
Feed Efficiency 4.33 3.90 0.430 0.015
38% alfalfa hay Nutrient % of DM
Figure 2: Effect of protected B vitamins blend Figure 3: Effect of protected B vitamins blend on daily gain
30% steam flaked corn Dry matter 88.4% on total gain and feed efficiency

10% corn distillers grain Crude Protein 18.5%

%
50
2.50 5.1
ADF 4.3%
2% soybean meal
NDF 9.6% 48 2.40 4.9
11% sugar cane molasses
Fat 3.2% 2.30 4.7
46
3% wheat straw NEg 1.0 mcal/kg 46.3b
2.20 4.5
2.20b
Ca 1.31%
Kg / day

6% minerals, vitamins 44 2.10 4.3

Kg

and soya 4.33a

P 0.38%
2.00 4.1
42 42.4a 2.01 a

1.90 3.9
Common diet for both groups (Figure 1). 40
1.80
3.90b
3.7
Animals weighed individually on day 1 and day 21. 38 1.70 3.5
Daily pen feed intakes measured.
Data analyzed as a randomized complete block design using pens as experimental Total Gain Average Daily Feed Efficiency
units. Gain ( Feed / gain)
Control Treatment
General linear model included treatment as fixed and block as random effects. a,b differs p<0.05

CONCLUSION
The addition of the rumen protected B vitamins blend in the diet improved the productivity of finishing beef cattle during the receiving period by improving weight gain, without
changing feed intake. Morbidity and mortality were very low in this facility, and there were no treatment differences in these parameters. Rumen protected B vitamins may be an
economical approach to improving productive performance in the feedlot.
Poster Jefo - 2015 - 30 X 40

REFERENCES
1. GIRARD, C.L. AND J.J. MATTE. 2006. Impact of B-vitamin supply on major metabolic pathways of lactating dairy cows. Can. J. Anim. Sci. 86:213220.
Poster available via Poster in my pocket application

2. SANTSCHI, D. E., R. BERTHIAUME, J. J. MATTE, A. F. MUSTAFA, AND C. L. GIRARD. 2005. Fate of supplementary B-vitamins in the gastrointestinal tract of dairy cows. J. Dairy Sci. 88:20432054 Code 1075e4a
EFFECT OF RUMEN PROTECTED B VITAMINS
ON FOLLICULAR DEVELOPMENT IN DAIRY COWS BY
MEASURING NUTRIGENOMIC PARAMETERS
Franois J. Richard1, Daulat R. Khan1, Christiane L. Girard2, Hlne Leclerc3,
Essi Evans3, 1Laval University, 2Agriculture and Agri-Food Canada 3Jefo Nutrition

From : Effects of a dietary supplementation of rumen protected B vitamins on reproduction of dairy cows by measuring nutrigenomic parameters, J. Anim. Sci Vol. 94, E-Suppl. 5/J. Dairy Sci. Vol. 99, E-Suppl. 1

Figure 2. Venn diagram showing number of differentially expressed transcripts between

OBJECTIVE protected B vitamins treatment and control group compared to 44 hours of coasting period
post FSH genes
To assess whether rumen protected B vitamins given as a dietary supplement can have an
impact on gene expression in granulosa cells as an indicator of follicular development.

MATERIAL AND METHODS 721 320 1259

262 1
30 multiparous cows were evenly divided in 3 treatment groups and housed at the AAFC 37 20
research station. Cows were fed a typical Canadian Total Mixed Ration (TMR). Treatments
are identified in table 1.
Table 1. Description of treatment Protected B vitamins FSH
Treatment - 21 days to calving 0 to 21 DIM 21 to 60 DIM 15 genes were validated through qRT-PCR and 11 genes expressed
Negative control No B vitamins differently for the protected B vitamins treatment compared to control or injection.
Injection1 of Folic acid Effect on oocyte quality (qRT-PCR)
Injection folic acid (320 mg) and B12 (10 mg)
T1 and B12
Diet-protected B vitamins Transition2 B vitamins Transition2 100 g/cow/day B vitamins Lactation3 > RGS2, INHBA, FOLR2 and NCR3C1 gene are well known markers for oocyte quality in
T2 B vitamins 50 g/cow/day B vitamins Lactation3 3 g/cow/day 3 g/cow/day human and bovine.
> Those 4 genes were upregulated in the protected B vitamins treatment and their expression
1
Weekly intramuscular injection was significantly different than control or injection treatment (figure 3).
2
Protected B vitamins for Transition: Folic acid, B12, Choline and Riboflavin 21 to 60 DIM
3
Protected B vitamins for Lactation: Folic acid, B12, Pyridoxine, Pantothenic acid and Biotin
Figure 3. Effect of protected B vitamins or injection treatment on differentially expressed
> Follicles size was measured by ultrasound from 40 DIM until Ovarian Pick Up at 54 DIM. transcripts coding for markers involved in oocyte quality. (validated by qRT-PCR)
> Cows were synchronized with two injections of PGF2alpha 12 days apart with first injection RGS INHBA
at 40 DIM. 6.0 x 10-6 5.0 x 10-4 8.0 x 10-5 5.0 x 10-6
b
b b b
> Granulosa cells were collected from dominant follicle (larger than 12 mm) 53 hours post 4.0 x 10-6
4.0 x 10-4 6.0 x 10-5 4.0 x 10-6
3.0 x 10-6
second injection of PGF2alpha. 3.0 x 10-4
5.0 x 10-4
a 4.0 x 10-5
2.0 x 10-6
a a
> Microarray, Ingenuity Pathway Analysis Software and RT-qPCR were used to identify, ana- 2.0 x 10 a
-6
2.0 x 10-5
1.0 x 10-6
a a 5.0 x 10-4
a a
lyze and validate gene expression. 0 0 0 0

Control Injections Protected B vitamins

RESULTS Results indicate that the dairy cows fed the protected B vitamins treatment showed a
potential for high oocyte quality.
Only cows having similar estradiol and progesterone level in follicular fluid were eligible
for analysis in order to have a same follicle physiological status. Effect on the follicle preparation for ovulation
1) Metabolic parameters: > RGS2, INHBA, OLR1, LHCGR, HSD3B1 and FST gene are well known markers for terminal
>The plasma folic acid was higher in the injection and protected B vitamins treatment differentiation which indicate that the follicle is preparing for ovulation.
compared to the control group. The level in the follicular fluid was also higher in the injection > OLR1 INHBA, RGS2 were upregulated in the protected B vitamins treatment and their
treatment than the control with no difference between protected B vitamins treatment and expression was significantly different than control or injection treatment
injection or control treatment. (figure 4)
>The B12 level in plasma and follicular fluid was higher in the injection treatment compared to
the control or the protected B vitamins treatment. Figure 4. Effect of protected B vitamins or injection treatment on differentially expressed
transcripts coding for receptors or markers involved in early response to LH. (validated by
> Neither the injection or protected B vitamins treatment did show a significant difference qRT-PCR)
from the control on any of the other parameters.
INHBA
The physiological status of the dominant follicle was not different between treatments 2.0 x 10-6 5.0 x 10-4
which confirmed that the nutrigenomic response allowed to measure the effect of b b
treatments. 1.5 x 10 -6 4.0 x 10-4 6.0 x 10-6
b
1.0 x 10-6
3.0 x 10-4 a 4.0 x 10-6
2) Nutrigenomic parameters: 5.0 x 10-7
5.0 x 10-4
a 2.0 x 10-6
a a 5.0 x 10-4
a a
Differentiation between protected B vitamins and injection treatment 0 0 0
> Less than 10% of the granulosa cell genes from the protected B vitamins treatment were Control Injections Protected B vitamins
common with the injection treatment.
The protected B vitamins treatment showed a different effect on the follicular > LHCGR, HSD3B1 and FST were downregulated with the protected B vitamins treatment
development than the injection treatment. and their expression was significantly different than control or injection treatment (figure 5).
Figure 1. Venn diagram showing number of differentially expressed transcripts compared to
control group between protected B vitamins treatment and injection treatment (microarray Figure 5. Effect of protected B vitamins or injection on differentially expressed transcripts
analysis) coding for receptors or markers involved in early response to LH. (validated by qRT-PCR)
LHCGR
1.5 x 10-6 1.5 x 10-4 a 4.0 x 10
7
a 3.0 x 10
8
1.0 x 10 -6
1.0 x 10 -4
a, b
a, b b b 2.0 x 10
948 93 162 5.0 x 10-7 5.0 x 10-5 1.0 x 10

3 0 0

75 Control Injections Protected B vitamins

Protected B vitamins Injections The upregulation and downregulation of cited genes indicate that the dominant follicle
from the cows fed the protected B vitamins was preparing earlier for ovulation, since
Effect on oocyte quality (micro arrays) they are pointing towards terminal differentiation of granulosa cells.
> A method used for high oocyte quality in superovulation condition consists of FSH
treatment followed by a withdrawal of FSH 44 hours before oocyte recovery (also called
coasting) (Nivet et al.,2012, 2013). CONCLUSION
> The granulosa cell genes of protected B vitamins treatment were compared to genes related The dominant follicle of the dairy cows fed the protected B vitamins blend
to 44 hours coasting period post FSH (44-post FSH). develop differently than the control group or the cows injected with folic acid
> 31% (320/1041) of the granulosa cell genes from the protected B vitamins treatment were and B12.The different response consists of an earlier preparation of the
common to the 44-post FSH follicle for ovulation and an improved oocyte quality.
> 93% (299/320) of the common genes were expressing similarly than the 44-post FSH
40 40.6b
38 38.1b
36
35.8a
34 34.0d
32 33.4a
30
29.9c
28

Control Protected B. Vit.

42 days after AL
949 multiparous Holstein cows Ref: Juchem, et al., 2012
 441P EFFECT OF A PROTEASE ON PERFORMANCE AND INTESTINAL HEALTH OF
BROILER CHICKENS FED A STANDARD DIET OR A LOW-DENSITY DIET
Mariana L. de Moraes1*, Ktia M. Cardinal2, Ines Andretta2, Elizabeth Santin3,
Derek Detzler1, Ludovic Lahaye1, Andrea M.L. RIbeiro2
1JefoNutrition Inc., Saint-Hyacinthe, Canada; 2Federal University of Rio Grande do Sul,
Porto Alegre, Brazil. 3Federal University of Paran, Curitiba, Brazil
*mmoraes@jefo.com

Introduction Table 2. Zootechnical performance

Standard Diet Low-Density Diet2

Protease enzymes can improve the dietary protein utilization. Therefore, it is possible to decrease the level of P-
dietary protein to save on feed cost, to improve performance, to reduce nitrogen excretion in the environment, No
Protease1
No
Protease
value
and to minimize the risk of enteric infections. The objective of this study was to evaluate the e ect of a Protease Protease
Protease on performance and intestinal health of broiler chickens fed a standard diet or a low-density diet. 1-7 days No protease Protease1
BW 7d, g 170 169 168 169 0.90
FI, g 166 162 172 165 0.80
BWG, g 120 120 118 120 0.88
FCR 1.37 1.35 1.45 1.38 0.55
Materials and Methods
2900 1.80
7-21 days
BW 21d, g 876a 848ab 802b 842ab 0.05 2,854x 1.76b
2850 1.75
> Location: Laboratrio de Ensino Zootcnico, Federal University of Rio Grande do Sul FI, g 1,014 1,027 1,006 1,030 0.79
BWG, g 706a 679ab 635b 673ab 0.04
> Animals: 392 day-old male broiler chickens (Cobb 500), 1-42 days of age, 28 floor pens (14 birds/pen) FCR 1.44a 1.51b 1.58b 1.53b <0.01 2800 1.70
> 4 treatments with 7 replicates/treatment. 21-35 days 2,769xy

Body Weight (g)

BW 35d, g 2,144 2,146 2,040 2,100 0.23

Adjusted FCR3
2750 1.65
FI, g 2,095 2,094 2,222 2,067 0.20 2,730xy
Table 1. Treatments distribution
BWG, g 1,268 1,298 1,237 1,258 0.68 1.61a
Feed Formulation Supplementation FCR 1.65a 1.61a 1.80b 1.64a <.001 2700
1.60a
1.60

35-42 days 2,664y

STD-P Standard Diet1 - 1.57a
BW 42d, g 2,730xy 2,854x 2,664y 2,769xy 0.10 2650 1.55
STD+P Standard Diet Protease3 (125 g/t) FI, g 1,010 1,194 1,121 1,100 0.27
LDD-P Low-Density Diet2 - WG, g 586y 709x 624xy 669xy 0.07
2600 1.50
FCR 1.72 1.69 1.79 1.64 0.20
LDD+P Low-Density Diet Protease (125 g/t) 1-42 days Standard Diet Low-Density Diet2
1Based on the Brazilian Tables for Poultry and Swine (Rostagno et al., 2011) divided in 4 phases: 1-7, 7-21, 21-35, and 35-42 d. BW 42d, g 2,730xy 2,854x 2,664y 2,769xy 0.10 2550 1.45
26% reduction in crude protein and amino acids. The ratio of the other essential amino acids to lysine was kept the same as in FI, g 4,284 4,477 4,521 4,361 0.26
Figure 1. Body weight (bars) and adjusted
the Standard Diet. BWG, g 2,680xy 2,805x 2,615y 2,719xy 0.10
3Jefo Protease, Jefo Nutrition Inc.
FCR 1.60a 1.60a 1.73b 1.60a
feed conversion ratio (lines) at day 42.
<.001
FRC (adj)3 1.61a 1.57a 1.76b 1.60a <.001
Evaluations:
1JefoProtease, Jefo Nutrition Inc.
> Zootechnical performance was evaluated by feeding periods. 26% reduction in crude protein and amino acids. The ratio of the other essential amino acids to lysine was kept the same as in the Standard Diet.
3Adjusted to 2.754 kg (average BW at 42d). Considers that there is a 0.3 increase in feed conversion for every 1 kg of body weight gain over 2.754 kg.
> Intestinal health: At 28 days, a sample of ileum was collected from two birds per replicate and analyzed by the BW, body weight; FI, feed intake; BWG, body weight gain; FCR, feed conversion ratio. Means followed by di erent letters di er (abcP<0.05, xyP<0.1).
I See Inside Scoring System Methodology (ISI), according to Kraieski et al. (2016). In this methodology, a
lower number of ISI index represents a better intestinal health.
Table 3. Histology analysis of the ileum - I See Inside Scoring A B
System Methodology (ISI)
Statistical analysis: ANOVA and Fischer LSD (GLM, SAS Inst. Inc.) for a completely randomized design.
Standard Diet Low-Density Diet2
P-
No No value
Protease1 Protease A B
Results
protease protease
Lamina propria
0.96b 0.76b 0.82b 0.37a 0.02
thickness
The reduction of 6% in dietary protein adversely a ected body weight (BW) and feed conversion ratio (FCR)
Epithelial thickness 0.07a 0.04a 0.21b 0.07a 0.02
of the chickens on the treatment not supplemented with the Protease (Table 2, Figure 1). . A B
Proliferation of
0.13ab 0.06a 0.23b 0.10a 0.04 C D
enterocytes
The supplementation of Protease on the Low-Density Diet allowed the recovery of performance losses due to Epithelial plasma
poor BWG and FCR, promoting a zootechnical performance similar to the control treatment (Standard Diet & infiltration
0.64 0.66 0.82 0.76 0.16
no Protease supplementation; Table 2, Figure 1).
Mixed inflammatory
infiltration in the 1.91 1.44 1.62 1.83 0.39
The supplementation of Protease on top of the Standard Diet tended to improve BW, treatment in which the lamina propria
E F
birds were 124g heavier and presented the same FCR at 42 days when compared to the ones fed the same diet Increase of goblet
formulation with no Protease supplementation (Table 2, Figure 1). 2.00 1.58 1.74 1.93 0.26 Figure 2. Ileum (Galus galus). A. Non treated
cells
Congestion 0.58 0.42 0.46 0.17 0.15 animal. Enlarged lamina propria in a fused villus.
The birds supplemented with the Protease presented the best ISI gut health index, and this result was mainly Necrosis / apical B. Treated animal. Reduced amount of fused villi
due to the e ect on lamina propria, epithelial thickness and proliferation of enterocytes (Table 3, Figure 2). 0.09 0.17 0.05 0.05 0.60 and more villi with thinner lamina propria. C. Non
karyolysis
treated animal. Observe the proliferated epithelial
Presence of oocysts 0.00 0.00 0.00 0.00 -
folds. D. Treated animal with less proliferated
Total 6.38b 5.13a 5.95ab 5.29a 0.06 epithelial folds. E. Non treated animal. Thicker
1Jefo Protease, Jefo Nutrition Inc. epithelium. F. Treated animal. Thinner epithelium.

Abstract
26% reduction in crude protein and amino acids. The ratio of the other essential amino
Hematoxylin and Eosin + Alcian blue stain; 100
acids to lysine was kept the same as in the Standard Diet.
abMeans followed by di erent letters di er (P<0.06). magnification.

We evaluated the e ect of a protease when supplemented on top of a standard diet or with a low-density diet
on the performance and intestinal health of broiler chickens. Male Cobb chicks (392; 1-42d) were reared in floor
pens and allocated in a completely randomized design in a 2x2 factorial with 7 replicates. There were 2 feed
formulations: a standard diet (STD) based on the nutritional recommendations of the Brazilian Tables for
Poultry and Swine (Rostagno et al., 2011) and a low-density diet (LDD), with 6% reduction in crude protein and
main digestible amino acids. The 2 diets were either supplemented (+P) or not (-P) with a protease (Jefo
Protease at 125 g/t). The performance was evaluated by feeding period (1-7, 7-21, 21-35 and 35-42d). At day 28,
samples of ileum of two bird/replicate were analyzed by a morphometric index for histological alterations (I See
Inside Scoring System - ISI). There was no interaction between factors and no di erences between the
treatments were observed in the 1-7d period. In general, for all the other periods, birds fed the LDD-P were
lighter and/or had poorer feed conversion ratio (FCR) when compared to all other treatments (P<0.05).
However, the supplementation with the protease on the LDD was able to a ect positively body weight gain
(BWG) and FCR (P<0.05) and to promote a performance similar to the birds fed the STD-P. At 42d, the birds
on the STD+P were the heaviest (124g di erence to STD-P, P=0. 1) and presented the same FCR of the
STD-P and LDD+P groups while being 13 points lower (P<0.001) than the LDD-P group. Regarding the gut
health analysis, the birds supplemented with the protease presented the best ISI morphological index (P=0.06)
mainly as a result of the lower number of alterations regarding lamina propria and epithelial thickness and
enterocytes proliferation. In conclusion, the protease tested improved performance and intestinal health
indicators of broiler chickens when supplemented on top of a standard diet or with a low-density diet.
KEYWORDS: broiler chickens, intestinal heath, histology, performance, protease.
Conclusions & Implications
Bibliography Kraieski AL; Hayashi RM; Sanches A; Almeida GC; Santin E. Effect of aflatoxin experimental ingestion and Eimeria lanitsetni The Protease studied can be used on top of a broiler standard diet to improve zootechnical performance or
no segnellahc eniccav histopathology and immune cellular dynamic of broilers: applying an Intestinal Health Index. Poultry Science. doi: 10.3382/ with a low-density diet as a strategy to reduce feed cost avoiding loss of performance. In both cases, the
ps/pew397. 2016. Rostagno HS; Albino LFT; Donzele JL; Gomes PC; Oliveira RF; Lopes DC; Ferreira AS; Barreto SLT; Euclides RF. Tabelas brasileiras
para aves e sunos: composio de alimentos e exigncias nutricionais de aves e sunos. 3edio, Viosa, MG: UFV, 252 p., 2011. SAS Institute Inc. 2011.
protease tested positively impacted indicators of intestinal health.
Base SAS 9.3 Procedures Guide. Cary, NC: SAS Institute Inc.
XYLANASE DORIGINE BACTRIENNE SUR DES RGIMES MAS / BL
EFFET SUR LES PERFORMANCES DE POULETS DE CHAIR
Bodin Jean-Christophe1, Boudry Christelle2,
1
Jefo, 2 rue Claude Chappe, Espace Performance La Fleuriaye, 44481 Carquefou Cedex,
2
Beldem, une division de Puratos NV, Rue Bourrie n12 - B-5300 Seilles (Belgique)
Contact : jcbodin@jefo.ca

INTRODUCTION RSULTATS
Lindustrie avicole requiert des optimisations de plus en plus prcises PERFORMANCES DE CROISSANCE
des formules, tout en prenant en compte les disponibilits et variations
de prix des matires premires sur les marchs mondiaux. 2800 2
Cependant, lutilisation nergtique des crales par le poulet peut 1,86x
tre altre par la prsence de hauts niveaux de polysaccharides non 2400 y 1,8
1,82
amylacs (PNA) solubles et insolubles (Dusel et al., 1998).
1,62a
2000 1,6
1,59b

Poids Vifs finaux (g)

OBJECTIF 1600 1,4

IC
1200 1,2
1,19a
bactrienne sur les performances de croissance du poulet de 1,17b

chair, recevant des formulations diversifies mas/bl. 800 1

400 0,8

0 298 302 966 1088 2420x 2525y 0,6

0-10j 10-21j 21-35j

Poids vifs IC Tmoin Essai

a-b (P < 0,05) ; xy (0.1 > P > 0.05)

Tmoin Essai p
MATRIEL ET MTHODES NOTATION
21 jours 3,20 3,60 <0,05 DE FIENTES
35 jours 3,10 3,35 ns
> 80 poussins mles Ross PM3 dun jour
> rpartition en 20 cages de 4 animaux de poids homogne
> identification individuelle Tmoin Essai p
> 2 traitements alatoires : RATIO 0 - 10 jours 2,57 2,49 ns
. tmoin ngatif EAU / ALIMENT
. tmoin ngatif + xylanase bactrienne (E 1606, 10 IU/kg) 10 - 21 jours 1,98a 1,87b <0,05
21 - 35 jours 1,83 1,76 ns
> 10 rptitions par traitement
> traitement des donnes via le logiciel SPSS
test Chi-Deux, test de rang (notation des fientes) CONCLUSION
Dmarrage Croissance Finition L tude dmontre que laddition dune xylanase dorigine
Ingrdient
(1-10j) (10-21j) (21-35j) bactrienne dans des rgimes diversifis sur base bl, mas et
Bl 31,40 34,00 35,00 tourteau de soja amliore la croissance des poulets :
Mas 30,00 34,00 36,00
Tourteau de soja 48 25,90 17,80 15,10 diminution significative du ratio eau / aliment
Tourteau de colza 3,50 5,50 3,60 amlioration significative de la qualit litire 21 jours
Drches de mas 3,00 3,00 4,00
Huile de soja 0,40 0,40 1,6
Prmlange CMV 5,80 5,30 4,70 GMQ*
Matire azote totale 20,00 237,5 205 + 4,4%
Lysine Digestible 1,05 0,91 0,82
Matires Grasses 2,61 2,78 4,36 IC*
Cendres Brutes 5,95 5,39 4,88 - 2,3%
* sur priode 0 - 35 j
Energie mtabolisable (Kcal/kg) 2801 2866 2999

RFRENCES:
Dusel, G., Kluge, H., Jeroch, H., 1998. J. Appl. Poult. Res., (7), 119-131
PosterJRA - 2015
IMPROVEMENT IN GROWTH PERFORMANCE,
INTESTINAL MORPHOLOGY AND GUT IMMUNITY
IN BROILERS SUPPLEMENTED WITH A SELECTED FORMULA
OF PROTECTED ORGANIC ACIDS AND ESSENTIAL OILS

Xin H , Yang X ,Yang C , Detzler D , Sary K , Bodin JC , Lahaye L

1 1 2 2 2 2 2
1
College of Animal Science and Technology, Northwest A&F University, Yangling, China
2
Jefo Nutrition Inc., Saint-Hyacinthe, (Qubec), Canada,

Sary K2

As the poultry industry is moving towards antibiotic-free rearing, many
alternative products offer an appealing compromise as a gut health promoter but
individual validation of their efficiency is required. This experiment was conducted to
compare growth performances and intestinal morphology in broilers supplemented
with a selected formula of protected essential oils and organic acids P(OA+EO),
an antimicrobial growth promoter or a non-medicated diet. A total of 450
chicks (Cobb 500) were randomly allocated to one of the three groups:
Control (non-medicated), Antimicrobial growth promoter (AGP) (enramycin at
ABSTRACT
150g/MT) and Feed additive (protected essential oils and organic acids at levels were significantly i n increased in duodenum in Feed additive group when
300g/MT). Feed to gain ratio compiled from 22 to 42 days from Feed additive group compared with Control and AGP groups (P <0.05). We concluded that the addition
was significantly better when compared with Control group (P <0.05) but did of a selected formula of protected organic acids and essential oils could improve
not demonstrate a significant difference when compared to AGP group (P> 0.05). nutrient utilizationefficiency in broiler for zootechnical performances, in association to
Feed additive group presented lower mortality compared to other groups, although improvement in intestinal morphology and increase in SIgA intestinal secretion,
not statistically significant. Morphometric measurements of jejunum villi from Feed positioning this product as an interesting alternative to antimicrobial growth promoters
additive group were significantly increased when compared with Control group (P in poultry production.
<005) but not statistically different when compared toAGP group (P <0.05). SIgA

INTRODUCTION RESULTS
Antimicrobial growth promoters administration in the poultry industry is progressively phasing out. Supplementing birds with a selected formula of protected organic acids and essential oils
This practice is bending to the increased public health concerns of resistant pathogenic bacteria P(OA+EO) :
transmission to humans and to the gratifying marketing image. Potentiate feed conversion possibly due to improvement in nutrient utilization
efficiency in broilers (Table 1);
Lower mortality;
properties and have been used in poultry nutrition, Yang et al., 2015. However, these compounds Improve intestinal morphology, maximizing absorption of nutrients (Table 2 and Figure 1);
activity maybe impaired in the foregut by the acidic pH and may not exert their full Increase SIgA intestinal secretion for protection of the mucosal surface against
properties in the hind gut. Protecting the selected organic acids and essential oils by a vegetable pathogens (Figure 2).
fat matrix ensure delivering these compounds to the hindgut by progressive digestion with
endogenous lipase and maximizing their benefits in the hindgut. Zootechnical parameters.
Table 1: Average adjusted daily gain (ADG) and Feed to Gain ratio (F:G) for each groups for 22 at 42 days
In addition to developing management skills during the transition towards antibiotic-free rearing, poultry
producers are demanding for products to aid in maintaining performances. Zootechnical parameters
Groups 0 to 21 days 22 to 42 days
Intestinal morphology changes are often investigated in order to monitor the impact of nutrition on gut ADG (g) F:G ADG (g) F:G1
physiology. In fact, increased intestinal villi height has been associated with greater absorption, Caspary Control 31.9 0.52 1.43 0.02 64.7 1.70 1.90 0.27a
1992. AGP 32.2 0.33 1.42 0.01 66.5 0.80 1.79 0.28b
P(OA+EO) 32.3 0.95 1.44 0.01 65.2 1.04 1.80 0.24b
The intestinal epithelium is constantly bombarded with external agressors, such as toxins and enteric Average P value obtained for F:G ratio is 0.006.
pathogens. Secretory immunoglobulins A (SIgA) interfere pathogenic microorganisms, therefore
participating to the mucosal immunity in the intestinal lumen, Mantis et al., 2011. Mortality. Feed additive P(OA+EO) group presented lower mortality (2.67%) at 42 days
and essential oils on SIgA levels are currently unknown. compared to Control (3.34%) and Antimicrobial Growth Promoter, (4.00%) groups,
although not statistically significant.

OBJECTIVES Intestinal morphology.

Table 2: Mean morphometric measurments of the villi and crypt from the jejunum for birds from each groups at 42 days

To evaluate, based on zootechnical performances, intestinal morphology and mucosal immunity, a Mean morphometric measurements of jejunum (m)
selected formula of protected organic acids and essential oils P(OA+EO) as a replacement to an Groups 42 days
antimicrobial growth promoter. villi height crypt depth wall thickness villi:crypt ratio
Control 805 54b 115 14 132 8 7.09 0.33ab
AGP 859 14b 99 3 140 17 8.64 0.33b
MATERIAL AND METHODS P(OA+EO) 955 42a 121 7 143 10 7.90 0.23a
Mean
a, b
P value obtained for villi
Experimental design. A total of 450 Cobb 500 chicks were randomly placed on 30 pens - 15 birds per pen, height is 0.012 and villi to crypt ratio is 0.02.

10 pens per groups.

Mucosal immunity.
Three groups were: Figure 2: Average quantity of secretory immunoglobulin A (SIgA) in each intestinal segment content per groups at 42 days
1. Control: non-medicated diet 13.24
2. Antimicrobial growth promoter (AGP): Control diet + enramycin at 150 g/MT 14
Quantity of secretory immunoglobulin (SIgA)

3. Feed additive P(OA+EO): Control diet + Protected organic acids and essential oils P(OA+EO) 12
at 300 g/MT 10 7.96
8.60
7.57 7.80
7.80
Birds were supplemented throughout the rearing period from 0 to 42 days.
(mg/100mg protein)

8.01a
8
6.48b
5.29c
Zootechnical parameters. Feed intake, body weight and bird mortality were recorded weekly per pen. 6
Control

Average daily gain (ADG) and Feed to Gain (F:G) were compiled at 21 and 42 days of age. 4
AGP

P(OA+EO)
2
Intestinal morphology. Medial portion of approximately equal portions of ascending and descending
0
jejunum samples were collected from 1 bird/pen (10 reps). Standard staining protocol with hematoxylin and Duodenum Jejunum Ileum

eosin was used. The mean villus height, crypt depth, wall thickness and villi to crypt ratio were measured by a-c
Average P value obtained for duodenum is 0.04.
3-trained personnel blinded of groups. Awad et al. 2008.
Mucosal immunity. The secretory immunoglobulin A (SIgA) levels in duodenum, jejunum and ileum were CONCLUSION
determined using a commercial RIA kit (SIgA RIA kit, China Institute of Atomic Energy, Beijing, China).
This blend of protected organic acids and essential oils P(OA+EO) offers a
Statistical Analysis. The data were analyzed by ANOVA test using the GLM procedure of SSPS as a promising solution for the replacement of antimicrobial growth promoters in
order to maitain birds performance and serve as a gut health modulator in the
further analyzed by Duncans multiple range test. P value was set at 0.05. current context of the poultry industry.

REFERENCES
AWAD W, GHAREEB K and BHM J. 2008. Intestinal Structure and Function of Broiler Chickens on Diets Supplemented with a
Synbiotic Containing Enterococcus faecium and Oligosaccharides. International Journal of Molecular Sciences. 9(11):2205-2216.
CASPARY, W. F. 1992. Physiology and pathophysiology of intestinal absorption. American Journal Clininal Nutrition. 55 (1 Suppl): 299S-308S.
MANTIS, J. ROL, N. and CORTHESY, B. 2011. Secretory IgAs complex roles in immunity and mucosal homeostasis in the gut. Mucosal Immunol. 4(6): 603-611. Jefo 2015
YANG, C. CHOWDHURY, M.A. HUO, Y. and GONG, J. 2015. Phytogenic compounds as alternatives to in-feed antibiotics: potentials and challenges in application. Pathogens. (4)1: 137-156. Abstract #: 425P
Session: Feed Additives
Location: Ballroom AB
 #446P Performance and intestinal microbial profile of broiler chickens
supplemented with a blend of protected organic acids and essential oils
Glenmer B. Tactacan*, Kathleen Sary , Wayne Bradshaw, and Derek Detzler
Jefo Nutrition, Saint-Hyacinthe, Quebec, Canada J2S 7B6

ABSTRACT INTRODUCTION
The increasing concern about the use of antibiotics in poultry production has changed the ways in which producers manage the birds The growth-promoting effects of antibiotics are strongly
overall health. Currently, additives with anti-microbial and growth promoting effects are added in poultry feeds to prevent and control related to their ability to modulate the gut microflora.
GI-tract infections that adversely affect performance. A study was conducted to determine the effects of a blend of protected Gut microflora significantly affects nutrition, health status, and
organic acids (OAs) and essential oils (EOs) in performance and intestinal microbial profile of broiler chickens. A total of 612 Ross animal performance by interacting with GI-tract development
308 day old chicks were randomly assigned to receive 1 of 3 treatments for 28 d: 1) basal diet with no antibiotic + 100 ppm lasalocid and nutrient utilization.
(T1) (n=204), 2) T1 + 300 ppm of protected OAs and EOs (T2) (n=204), and 3) T1 + 1500 ppm of protected OAs and EOs (T3) Therefore, the focus of alternative strategies to antibiotics has
(n=204). A completely randomized design with 3 treatments, 12 replicates, and 17 birds in each replicate was used. On d 14 and 28, 1 been centered on modulation of the gut microflora, in particular
bird from each pen was sacrificed to collect ileal and cecal samples for microflora analysis using high-throughput sequencing based the prevention of proliferation of pathogenic bacteria in the gut.
on 16S rRNA genes. The BW of birds in T2 and T3 at d 21 was significantly increased relative to T1 (P<0.02, 4.6%), as was the BW of

OBJECTIVES
birds in T2 at d 28 (P<0.05, 2%). The FCR was not different between treatments; however, there was a trend towards improved FCR
at d 21 in T2 (P<0.09, 5.5%) and T3 (P<0.06, 5.6%), as well as at d 28 in T2 (P<0.06, 5.7%). Sequencing data at d 14 and 28 revealed
retained complexity and overall structure of the ileal and cecal microbiota across treatments. However, the intestinal microbial profile
of treatments changed in between these time points. Compared to T1, significant changes in the abundance of some Lactobacillus The study aimed to demonstrate the impact of supplementing
species within the cecum of birds in T2 and T3 were found at d 28. Overall, the supplementation of a blend of protected OAs and protected organic acid and essential oil blend in terms of:
EOs had no adverse effect on the microbial diversity of the intestine and appears to offer benefits with respect to gut health and Gut microbial profile and,
productivity in broiler chickens. Indices of growth performance in broilers

MATERIALS AND METHODS Lactobacillus in Cecum @ D28

612 Ross 308 day old chicks in 36 pens were fed 3 di erent Microflora profiles were assessed and compared using OTU Identity
Homology
%
% in T1 % in T2 % in T3

rations: QIIME software. 76727 Lactobacillus salivarius 100 0.062 0.076 0.22
57808 Lactobacillus salivarius 100 1.13 1.35 3.57
Basal diet + 100 ppm lasalocid (T1) Operational Taxonomic Units (OTUs) represents a cluster 35274 Lactobacillus salivarius 97 0.018 0.019 0.096

T1 + 300 ppm of protected OAs and EOs (T2) of related DNA sequences at 97% sequence identity 217021 Lactobacillus salivarius 97 0.076 0.14 0.26

accepted as equivalent to species 196333 Lactobacillus salivarius 97 0.078 0.076 0.18

T1 + 1500 ppm of protected OAs and EOs (T3) 157835 Lactobacillus salivarius 98 0.076 0.087 0.17
STATISTICAL ANALYSIS 5797 Lactobacillus sp. crispatus/acidophilous/gallinarum 95 0.18 0.26 0.87
Feed intake and BW were measured weekly until d 28. 52716 Lactobacillus reuteri 96 0.063 0.061 0.21
Growth performance data were subjected to ANOVA.
On d 14 and d 28, 1 bird per pen was removed and samples of 35461 Lactobacillus kitasatonis 97 0.010 0.044 0.13
37371 Lactobacillus satsumensis 97 0.050 0.042 0.14
ileum and caecum were analysed for microbial profile analysis 2-tailed students t-test was used when ANOVA indicated 96403 Lactobacillus salivarius 95 0.093 0.15 0.45
using high throughput sequencing of 16S rRNA. significant di erences between groups Average Lactobacillus N/A 0.167 0.210 0.572

DIVERSITY MEASURES & DIFFERENTIALLY ABUNDANT OTUs

NO DIFFERENCES IN THE DIVERSITY OF CECAL MICROBIOTA INCREASED LEVEL OF LACTOBACILLUS SPECIES IN THE CECUM @ D28
5
OTU P=0.585 (CECUM @ D14) OTU P=0.413 (CECUM @ D28) 1.5
CECUM @ D14 (P < 0.05) CECUM @ D28 (P < 0.05)
Relative Abundance

4 *
Relative Abundance

*
Evenness Index

*
Evenness Index

1
3

* *
* 2
0.5 * *
** * * * * * ** * *
** ** * * * * *** * * ** * * * * * ** *
* * * *
* * * 1
T1 Control T2 300 ppm T3 1500 ppm T1 Control T2 300 ppm T3 1500 ppm * * *
OA + EO OA + EO OA + EO OA + EO
0 *
0
OTU P=0.321 (CECUM @ D14) OTU P=0.428 (CECUM @ D28)
Richness Index

Richness Index

T1 - Control T2 - 300 ppm OA + EO T3 - 1500 ppm OA + EO

T1 Control T2 300 ppm T3 1500 ppm T1 Control T2 300 ppm T3 1500 ppm
OA + EO OA + EO OA + EO OA + EO

GROWTH PERFORMANCE @ D21 & D28 CONCLUSION

BODYWEIGHT AND
BODYWEIGHT ANDFEED
FEED CONVERSION RATIO
CONVERSION RATIO @ D21@D21 BODYWEIGHT AND
BODYWEIGHT ANDFEED CONVERSION
FEED CONVERSION RATIORATIO
@ D28 @D28 > The use of microflora analysis is very helpful in confirming the
0.996b
impact of protected organic acids and essential oil blends in
1.330
1.02 0.984b 1.360 1.74
1.707b
1.8 poultry
1.719
Feed Conversion Ratio

1 1.72
> The supplementation of protected organic acids and essential
Feed Conversion Ratio

1.320 1.75
1.683ab
Bodyweight (kg)

0.98 0.944a 1.7 oil blends:

Bodyweight (kg)

1.261 1.259 1.656 1.7

0.96 1.280 1.657a 1.627
1.68 Had no adverse effect on the gut flora diversity
0.94 1.65
1.66
1.240 Increased the proportion of Lactobacillus spp in the gut
0.92 1.64 1.6
0.9 1.200 Appears to offer benefits with respect to gut health and
T1- CONTROL
T1 CONTROL T2
T2- 300 PPMPPM
300 OA + T3
T3- 1500 PPMPPM
1500 OA +
1.62 1.55 productivity in broiler chickens
T1 T1
-CONTROL
C ONTROL T2
T2- 300
300 OA +
PPMPPM T3
T3- 1500 PPM OA
1500 PPM+
OA EO+ EO OAEO+ EO
OAEO + EO OAEO+ EO > These results are supported by studies being reported
Bodyweight
Bodyweight Feed
FeedConversion
Conversion Bodyweight
Bodyweight Feed
FeedConversion
Conversion internationally
FREQUENCY OF AVIAN PATHOGENIC
E. COLI (APEC) IN BIRDS SUPPLEMENTED
WITH ANTIBIOTIC ALTERNATIVES
Dr. Kathleen Sary Jefo Technical Services
Stphane Benaben Jefo Innovation and Development

BACKGROUND RESULTS
While reducing antimicrobial usage, producers can Presence of APEC is lower in broiler feces receiving
experience higher mortality rates during the brooding P(OA+EO) than in any other feed additive or
and growing period. One of the causative agents for combination of.
this mortality is avian pathogenic Escherichia coli
(APEC), which can be identified by the virulence Table 1: Frequency of colonies with E. coli virulence
genes it contains in order to colonize extra-intestinal genes detected in feces of broilers.
sites.
No. of Frequency of APEC virulence genes 1
Groups colonies
OBJECTIVE tested iucD 2 tsh papC None 3

To compare the virulence of APEC found in broiler Control 30 24 11 4 3

chicken feces at 14 days when birds receive a Jefos
P(OA+EO)4 30 16 6 0 14
Protected Organic Acids + Essential Oils
P(OA+EO) or protected feed additives carvacrol, P(OA+EO)/C5 27 18 5 1 9
and benzoic acid.
Carvacrol 27 25 12 2 2

Benzoic acid 30 21 6 1 8

1
No tested colonies possessed the cnf virulence gene.
2
Virulence gene iucD presence gives 90 to 100% confidence
that it is an APEC.
3
Absence of the four virulence genes tested
MATERIALS AND METHODS (iucD, tsh, cnf, papC)
A total of 1440 Ross 308 chicks were randomly 4
Thymol based P(OA+EO) 5 Carvacrol based P(OA+EO)
allocated to pens corresponding to one of the four
treatments (10 replicates). Diet formulation follows CONCLUSION
standard Canadian broiler diets.
Jefos Protected Organic Acids + Essential Oils offers
T1 - Control T2 - P(OA+EO) T3 - P(OA+EO) T4 - Protected T5 - Protected
(not treated) 300g/t + carvacrol 300g/t carvacrol 4.8g/t benzoic acid 200g/t a promising solution to diminish avian pathogen
E. coli (APEC) challenge during the rearing period.
Fecal samples were collected at day 14 and presence
of the four most common APEC virulence genes
(iucD, tsh, papC, cnf) were analyzed by PCR at the
OIE reference Laboratory for Escherichia coli
(Quebec, Canada).
PERFORMANCES ET MICROBIOTE
INTESTINAL DU POULET DE CHAIR
EFFET DE LUTILISATION DUN MLANGE
DACIDES ORGANIQUES ET DHUILES
ESSENTIELLES MICROENCAPSULS
Sgolne Roche (1), Kathleen Sary, Peter Scott, Tim Wilson
1
JEFO Europe, 2 rue Claude Chappe, 44481 Carquefou

Contact : sroche@jefo.ca

OBJECTIF RSULTATS
Lobjectif de ce travail est d tudier limpact dune solution base
Graphique 2. Analyse de labondance relative des bactries caecales 28 jours
dacides organiques et dhuiles essentielles microencapsuls,
(P<0,05)
spcifiquement conue pour la volaille (AOm+HEm), sur les 5
Tmoin
performances de croissance et le microbiote intestinal de poulets AOm+HEm 300 ppm
de chair, selon deux niveaux de doses. 4 AOm+HEm 1500ppm

Abondance relative (%)

MATRIEL ET MTHODES 3
abc
612 poussins dun jour Ross 308 ont t rpartis en 3 traitements 2
de 12 rptitions :
1. Tmoin baa aab
aab
1
2. AOm+HEm 300 ppm : Tmoin avec AOm+HEm une aab abc abc aac abc
aab
aab aab aab
dose de 300 ppm b a ab a a
baa baa
0
3. AOm+HEm 1500ppm : Tmoin avec AOm+HEm une Lactobacillus salivarius
dose de 1500ppm

sa e
tri es
los es

les

ct um

ula
riu acea
los ut
C dial
dia

ec
tri
C ric

Bi ata ac
tri

ac eri
ne

Le suivi de la consommation daliment et des poids a t ralis

m
C alib
Te

a
ec

b
te
hebdomadairement jusqu 28 jours.

Fa

ob
fid
14 et 28 jours, un oiseau de chaque parquet a t prlev pour
ncropsie et chantillonnage du contenu caecal et de la Augmentation de labondance des lactobacilles et diminution
muqueuse de lilon. Les chantillons ont t analyss pour les des clostridiales dans les caeca 28 jours avec AOm+HEm
profils du microbiote par squenage de lARNr 16S. 300 ppm et 1500 ppm. Labondance des lactobacilles est plus
Les profils du microbiote ont t tablis et compars avec le logiciel importante avec AOm+HEm 1500 ppm.
QIIME.
Les OTU (Operational Taxonomic Unit) reprsentent un
regroupement de squences dADN lies. Un seuil de 97%
dhomologie permet destimer comme quivalent un OTU une
espce bactrienne Lanalyse du microbiote est un bon outil pour confirmer
ANALYSES STATISTIQUES : ANOVA, test de Tukey, variance limpact de la solution compose dacides organiques et
dhuiles essentielles microencapsuls, spcifiquement conue
pour la volaille.
RSULTATS
La supplmentation de AOm+HEm sur les poulets de chair :
Graphique 1. Poids et indice de consommation 28 jours (P<0,05) >
> augmente la proportion de lactobacilles et diminue
1,75 les clostridiales dans les caeca 28 jours, en parallle
1,719
1,70 dune amlioration des paramtres de productivit (avec
AOm+HEm 300 ppm).
1,65 1,627 1,656
1,60 Comme dj dmontr dans de nombreuses tudes,
AOm+HEm participe lamlioration de la productivit de la
1,55 1,707 b
1,683ab filire volaille.
1,657 a
1,50
Tmoin AOm+HEm AOm+HEm
300 ppm 1500ppm

Poids moyen (Kg) IC

Poster JRA - 2017

Amlioration des performances de croissance 28 jours avec

AOm+HEm 300 ppm.
 PERFORMANCE RESPONSE TO DIETARY SUPPLEMENTATION
OF PROTECTED BUTYRIC ACID AND A COCKTAIL OF PROTECTED
ORGANIC ACIDS AND ESSENTIAL OILS IN LAYING HENS
G. B. Tactacan*, S. Benaben, K. Sary, L. Lahaye, J. C. Bodin, and D. Detzler
Jefo Nutrition Inc., Saint-Hyacinthe, Quebec, Canada.

ABSTRACT RESULTS
The use of antibiotics in egg-producing hens may transfer drug resistant traits to Figure 1.
pathogenic bacteria as a result of drug residues left deposited in fully developed
eggs. Presently, organic acids and essential oils are two of the most commonly Rate of egg production of laying hens fed protected butyric acid and a cocktail
use additives in lieu of antibiotics in animal production. To this end, a study was of protected organic acids and essential oils: main effects of diet
Egg Production (P < 0.09)
alone or in combination on laying hen performance during the late stage of their 90,0
production. A total of 252 80-week old Lohmann Classic were randomly assigned 85.9
88,0
to receive 1 of 3 dietary treatments for 8 weeks: 1) basal diet with no antibiotic
86,0
(T1) (n=84), 2) T1 + 1000 g/ton protected butyric acid (T2) (n=84), 3) T1 + 81.5
84,0 80.9
300 g/ton protected organic acids and essential oils (T3) (n=84). A completely

(%)
+6%
randomized design consisting of 3 treatments, 21 replicates, and 4 hens in each 82,0
improvement
replicate was used. Performance parameters including percentage hen-day egg 80,0
production, feed consumption, egg weight, egg mass, and feed conversion were 78,0
measured. Data were analyzed using the mixed model procedure with compound 76,0
T1Control
- Control T2P(Butyric
- P (Butyric acid) P(OA
acid) T3 - P(OA+EO)
+ EO)

Figure 2.
dietary treatments. However, a trend towards improved egg production (P<0.09,
5.8%) and egg mass (P<0.09, 5.1%) were observed in T3 birds relative to T1 and Egg mass of laying hens fed protected butyric acid and a cocktail of protected
organic acids and essential oils: main effects of diet
day production (P<0.03), feed consumption (P<0.001), egg weight (P<0.001),
egg mass (P<0.02), and feed conversion (P<0.001). However, no significant Egg Mass (P < 0.09)
59,0 56.9
58,0
acids and essential oils tended to improve laying hen performance relative to 57,0
56,0 53.9 54.1
55,0 +5%
(g)

laying performance caused by the changes in laying hens age. improvement

54,0
Keywords: Organic acids, essential oils, butyric acid, performance, laying hens 53,0
52,0
51,0
INTRODUCTION T1Control
- Control T2 - P (Butyric
P(Butyric acid) P(OA
acid) T3 - P(OA+EO)
+ EO)

Relatively few antibiotics are labelled for egg-producing hen due to concern Table 1. Performance summary of laying hens fed protected butyric acid and a
of drug residues in chicken eggs
Following treatment or exposure to antibiotics, many weeks may be required
before eggs are free of drug residues due to the prolonged nature of egg Egg Feed Egg Weight Egg Mass
Week FCR
Production (%) Consumption (g/d) (g) (g)
development
1 85.7a 112.1ab 66.3abcde 56.9a 1.99cd
Therefore, natural based alternative to antibiotics are being used to prevent
2 84.5ab 105.7c 66.0de 55.8ab 1.90de
3 81.9bc 110.3b 66.3abcde 54.3bc 2.08abc
4 83.1abc 115.5a 66.8ab 55.5ab 2.12a
OBJECTIVES 5 83.0abc 103.1c 67.0a 55.6ab 1.87e
6 82.1bc 111.5b 66.7abc 54.7abc 2.07abc
organic acids and essential oils on metrics of laying hen performance 7 80.3c 102.9c 66.3bcde 53.2c 1.96de
8 77.0d 98.7d 65.9de 50.7d 2.00bcd
MATERIALS AND METHODS SEM 1.3 1.5 0.3 0.9 0.04
P-value 0.03 0.001 0.001 0.02 0.001
252 80-week old Lohmann Classic were randomly assigned to receive 1 of
3 dietary treatment for 8 weeks
CONCLUSION
> Treatment 1 = Basal diet with no antibiotic (T1), (n = 84)
> Treatment 2 = T1 + 1000 g/ton protected butyric acid (T2), (n = 84) The dietary supplementation of protected organic acids and essential
> Treatment 3 = T1 + 300 g/ton protected organic acids and essential oils
(T3), (n = 84) performance relative to protected butyric acid
Feed consumption, egg production, egg weight, egg mass, and feed
conversion ratio were measured weekly
Statistical Analysis
Completely Randomized Design (3 treatments, 21 replicates, 4 hens replicate)
Mixed model procedure with compound symmetry variance structure for
repeated measurements
PosterJefo - 2016

P < 0.05 was considered significant