Note: Descriptions are shown in the official language in which they were submitted.
~ W093/0620~ PCT/~IS92/07~X')
2116~25 -
FATTY ACID MICROENCAPSULATED ENTEROCOCCUS
FOR USE WITH POULTRY
. ,
BACKGROUND OF T~E INVENTION
Growth enhancers in the form of antibiotics have
been used extensively for poultry, namely chickens
and turkey. Growth enhancers su~h as Stafac ~ and ~.
BMD ~ ~bacitracin methylene disalicylate) are known
antibiotics and have been used at sub-therapeutic
levels of for example, 10 grams per ton and 25 grams
per ton as feed additives in order to promote
desirable growth features in poultry. However, the
use of antibiotics for these purposes has recently
come under some criticism. One of the criticisms is
the possibility that the poultry eventually develop
tolerance to the antibiotics and eventually the
antibiotic no longer works well for growth promotion.
Other objections relate to health concerns from non-
natural antibiotic additives and the adulterating
effects they may have. Nevertheless, because of the
advantages of antibiotic uses they are still commonly
use~ in order to improve feed conversion, improve
carcas:s composition, and enhance growth.
It is known that certain bacteria are
potentially beneficial when added t5 animal feeds.
These ~acteria are beneflcial in that they supply a
natural intestinal micro-flora. Some companies offer
for sale probiotics which contain desirable
ba~teria. Probiotics, however, do have some dif-
iculty in maintaining a s~able product. Typically,
the probiotic is used at a fairly low level, added
to feed at perhaps a 0.1% level. However, unused
probiotic containing feed or feed additive product is
often stored by the farmers for long periods of
time. This storage many times is under conditions
where there is some moisture and high temperature.
.,,
W093/06208 PCT/~iS92/07~
2 t 1 b ~ 2 ~
In many instances there is just enough moisture that
the bacteria are activated or start to grow, but yet
there is an insufficient amount of moisture to . .
sustain them. As a result they die. Thus, the
acti~ity of the probiotic is stopped. In other
instahces, ~he addition of antibiotics to the
probiotic containing feed or feed additive adversely
interacts wi~h the bacteria, particularly if there
are small amounts of moisture present and thus again
bacteria are killed. Thus, there is a significant
problem of long term storage stability for
probiotics.
In another environment, where the probiotic is
added to, for example chicken eed, it is common to
pelletize the material with the probiotic added
before pelletizing. Moisture from steam used during
pelletization partially activates the bacteria, but
may, as a result of insufficient moisture to sustain
them, kill them. ~Also heat during pelletization may
kill them. Then, too, there is the problem of the
acid environment of the stomach potentially
i~activating bac~erla before they really reach the
intestine. Thus, ~here is a continuing need for
pr~bio~ics which~will release the organisms only at
the proper time in the intestine, without early
release due to moisture conditions or adverse pH
conditions such as exist in the digestive tract
anterior to the small intesti~e.
Certain features of poul~ry are especially
de~irable ~o achieve if possible. Those include an
increased rate of weight-gain, better feed
conversion, carcass composition, and finally
uniformity of flock weight. Increased rate of weight
gain and better feed conversion are, of course,
desirable for the attendant economics ~hat accompany
. W~93/06208 PCT/~IS92/075~'3
2116S2~
3 --
these desirable results. The composition of carcass
is impor~ant because the most desirable area for
tissue deposit is the breast in order to yield a hi~h
amount of choice meat. Thus, weight gain is not only
important, but where the weight is gained on the
carcass ~s also important. Vniformity of flock
weight ls important because if more birds are normal
in size, less hand labor is required and processors
j can more extensively rely on mach~ne processing. On
the other hand, if the birds vary considerably from
very small birds to very large birds, even though the
; overall flock weight may be the same, the smaller
birds and the larger birds require a great deal more
31 hand labor and ~ecause of their lack of uniformity insize, ¢annot be processed easily by machine. Thus,
~ uniformity of flock weight wi~h a high percentage
`~ distribution within the normal size range so that
t chickens can process by standardized machinery is a
.3 desirable feature.
`: : It is a~primary objectlve of the present
l invention to provide a poultry probiotic which
'3 ' contains no antiblotics and contains only fatty acid
i
~ - microencapsulated:naturally occurring organisms.
, . .
~3 It is another primary objective of the present
invention to provide a probiotic which contains two
~ organisms, nameIy Enterococcus faecium 301, DSM No.
3 DSM-Nr. 4789, and Enterococcus faecium 202, DSM No.
~, I DSM-Nr. 4788. DSM is a Bacterial Culture collectio~
~ in Germany. DMS stands for Deutsche Sammlung von
,: Mikroorganismen located in Braunschweig, West
Germany. These organisms will be deposited at the
ATCC, with ~11 restrictions lifted upon notice of
, /
.~: allowable claims.
L~ It is a further objective of the presen~
-~ invention to provide a probiotic which, for poultry,
..:
"~
,
-
.,
W0~3/V620~ 2 1 1 6 S 2 S PCT/~iS92107~X'~
provides increased rate of weigh~ gain, which
provides better feed conversion, which provides
higher yield of breast meat, and which provides for~
uniformity of flock weight within the range of normal
slze.
An even further primary objective of the prese~t
invention is to provide probiotics suitable for
poultr~ feed ration addition which contains bacterla
that are in microsphere form using a special rotary
technique using free fatty acid matrix.
Another objective of ~he present invention is to
provide a probiotic which has stability at levels
within the range of from 3 months to 6 months without
any significant organism count reduction.
~ nother objective of the presen~ invention is to
provide a process of rotary formation of spheres of
the dried bacteria which provides having uniform
size.
Another ob~ective of the present invention is to
provide rotary disc spheres of dried bacteria which
are free flowing, and easily processable with poultry
feed rations. -
BRIEF DESCR~PTION OF THE DRAWINGS
Figures 1, 2 and 3 show graphically thestability of the strains using stearic acid ma~rix.
Figure 4 i5 a graph showing breast yield
distribution for a feeding trial of the probiotic
composition of the present invention.
Figure 5 is a graph showing body weight
distri~ution for a feeding trial o~ the probiotic
composition of the pr~sent invention.
Figures 4 and 5 show a control, use of an
antibiotic and use of the pro~iotic of the present
lnvention.
,
... .
.
W~93t0620~ P~T/~'S92/075X')
211~5~
-- 5
~UMMARY OF THE I~VENTION
The invention is a method and composition of
growth promotion for poultry which comprises adding
to the normal poultry feed ration a small but growth
promoting effective amount of a probiotic which
contains dried, atty acid microspheres of
Enterococcu$ aecium 301, DSM No. DSM-Nr. 4789, and
dried fat~y acid microepheres of Enterococcus faecium
202~ DSM No. DSM-Nr. 4788, where preferably the fatty
microspheres are formed by rotary disc drying.
DÉTAILED DESCRIPTION OF THE INVENTION
It has been surprisingly disco~ered that the
growth promotion of poultry can be accomplished by
adding to normal poultr~ feed rations, a certain
amount of fatty acid microspheres of F.n~erococcus
faecium 301, ~SM No. DSM-Nr. 478~, and a certain
amount ~f fatty acid microspheres of ~nterococcus
i fae~ium 202, DSM No. DSM-Nr. 4788. A fatty acid
t employed may be ~ny one of the C12 to C24 free fatty
acids, but is preferably stearic acid. The organisms
are preferably present in about equal amounts but may
vary within the range from about 30% to about 70~ of
one of the organisms with the balance being the
' o~her.
'`3! ' It is not known precisely why these two
-'~ organisms provide the desirable features of the
~ present invention, namely increased rate of weight
:. gain, better feed conversion, increased yield of
breast ~eat, and increased uniformity of flock
weight. The fact is that they do, provided that both
, are used in combination so that ~hey can somehow
'~J in~eract with each other, and providing that they are
-S used within the range herein expressed. It is these
, combinations of features which some how interact and
,.
.;,
~,
W093/0~08 PCT/~'S92/075~)
2116~2!ï
-- 6
co-act to provide the desirable features of the
present invention whlch allow significantly improved
poultry carcass, meat quality and processing.
The amoun~ of probiotic added to the feed ration
can vary considerably but generally will be within
the range of from about 0.5 pounds to about 2.0
pounds per ton of feed, generally from about 0.8
pounds to about 1.2 pounds per ton of feedj and
typically at ~bout 1 pound per ton of feed. The
organism count, that is the number of colony forming
units per gram present in the probiotic can also vary
within the range of from about 1 x 10 CFU/gm to
about 2 x 1~9 CFU/gm, but is preferably at about 2 x
CFU/gm.
. When ~he probiotic as previously described is
free choice fed in poultry feed ration, the
" .
i combination of two strains of organisms herein
mentio~ed, behave as a growth promoter. Growth
promoters now used include antibotics such as
i Stafac ~ and BMD. The advantages of sub-therapeutic
~: levels of antibioti~s as growth promo~ing additi~PS
. can be achieved with na~urally occurring organisms of
~t~ the present invention provided that probiotic is ~ade
`. in acco~dance~with ~he present invention and added in
i~ accordance with the me~hod described hereinO In
fact, thare have been some trials that suggest that a
" combination of probiotic and growth promotant
together exceeds the advantages of either alone and
~-~ thus they may be used together if desired. However,
~ in most instances, it is preferred to use the
.; probiotic alone since one of the objectives of the
`~ present in~ention is to avoid use of growth
Z promotants altogether.
:.~ The method of processing of the organisms is not
- critical as long as the organisms can be kept alive
. q
-.
,~
. W093/06208 PCT/~S92/07~
2116~5 -
-- 7
to delivery to the animal, and placed in a foxm so
that it will combine with animal feed well and is of
a generally uniorm size so that dosage may be
controlled.
A preferable means of achieving these
requirements is by providing the organisms in a
microsphere of a fat~y-acid ma~rix. This process if
described in the parent application of the co-
lnventor Rutherford, Pt al. By this process, the
bacteria are combined with a heated atty acid. The
temperature of the fatty acid and time of exposure of
the ~acteria to the fatty acid is controlled to keep
the bacteria alive, yet allow mixing with the fatty
acid. The mixture is placed on a rotating rotary
disk, with the result being a microsphere of bacteria
with a atty acid acting as the matrix. Several
important advan~ages are achieved using this method.
First, the bacteria are kept alive through the
processing; second, the process combined with the
rotary disk technlque~allows for a uniform size of
the microsphere for improved dosing. Third, the
nature of the matrix, a ~atty acid, allows ~he
fcrmatîon of the unique microspheres. The
combination of the f ctors provides for a highly
stable probiotic with maximum effectiveness~
In the process of the parent application it is
important to note mic~ospheres are formed wherein
each sphere consti;tutes a plurality of bacteria in a
free fatty acid matrix rather than an individual
microencapsulator of~each bacteria in a coating or
film like layer of fatty acid. This provides
stability advantages, and more effective dosing with
th bacterial treatment.
The preferred encapsulating agent is a C12 to
C24 free fatty acid. While mixtures of fatty acids
.'
W093/06208 PCT/USg2/075X'3
2:~C~2~ `
- 8 -
may be employed, it is preferred that a single pure
free fatty acid be employed. It is also preferred
that the free fatty acid be an unsaturated fatty
acid, with the most preferred being stearic Acid.
Generally ~peaking, it is important that the
fatty acid have a melting point less than 75~C,
preferably within the range of 40C to 75C. It
must, of course, be solid at room temperature in
order to be an effective matrix. All free fatty
acids falling within the range of chemica1
description heretofore given will meet these
requirements.
In order to enhance the product stability, the
bacteria are typically freeze-dried bacteria as
placed in the product. Thus, they can be revi~ed by
moisture addition.
In the microsphere, made in accordance with the
proce~s discussed below, the microspheres generally
compr~se from about 50% to over 90% by weight of ~he
fatty acid component with the balance being bacterial
culture. The preferred range is from about 60~ to
about 75% fatty acid. If too little fatty acid is
used, the matrix wlll be inadequate for protection.
On the other hand, if too much is used, the matrix
will be too thick and results in inadequate release
in the gut.
.. . .
The process as used in this invention is a
rvtary disc microsphere forma~ion process. Generally
I speaking in the rotary disc technology, a slurry of
tre bacteria and fatty acid components are thoroughly
mixed with the mixture being added at a uniform rate
i onto the center of a rotating stainless steel disc.
It is there flung outwardly as a result of
centrifugal force and forms a microsphere. It is
then collected in a cooling cham~er maintained at
r: .
. W093/06208 PCT/~'S92/075X'~
2116~25
g
ambient conditions or slightly lower, sized and
readied or packaging.
While ro~ary disc encapsulation is known, it is
not known to ma~e microsphere contained in a matrix
without a surrounding shell, nor is it known to use
the microsphere process or encapsulation with freeze
dried bacteria. Generally speaking, for descriptions
of rotary disc encapsulation, see a paper by Johnson,
et al. of the Southwest Research Institute of San
Antonio, in the Journal of Gas Chromotoqraphv,
October, 1965, pages 345-347. In addition, a rotary
disc encapsulator suitable for use in this invention
is described in detail in ~nited States Letters
Patent, Sparks, 4~675,140, issued June 23, 1987 and
entitled "Method For Coating Particles For Liquid
Droplets" the disclosure of which is incorporated
herein by reference. However, it is the process
dscribed in the parent ~hat is most preferred.
It is imp~rtant to note that rotary microsphere
formation provides a distinctly different product
than either conventlonal tower spray drying or
microencapsulation. In conventional tower spray
drying ther~ is a tendency for particles to cluster,
for the coating to be uneven, and thus for the
.
stability of.~he product to be significantly
effected perhaps from days ~o weeks.
Microencapsulation provides a shell coatin~ around an
obJect, and bacteria have proven to be too small, too
hard to keep alive or provide in a ~niform size to be
of practical usefulness. With microsphere formation,
particularly with agen~s used in ~his invention is
used, the stability of the resulting bacteria, even
when subjected to some moisture and antibiotics,
will be for from three to six months with the
viability of the bacteria maintained in evenly
distributed particles.
.
^,~'
W093/06208 PCT/~'S92/075X~t
2116525 lo~ -
When the ~ree fatty acid microspherss of the
present invention are used within the ranges
hereinbefore expressed, the rotary disk, typically
employing a 41'-6" rotary disc, can be run at the rate
of from 20~0 rpm to 4000 rpm, prefer~bly about 2500
rpm to 3200 rpm with a feed rate of from 50 grams to
200 grams per minute. The preferred conditions pres-
ently known are use of stearic acid, use o two
hereinbe~ore descrlbed organisms, a four inch rotary
disc, 3000 rpm and a feed rate of 100 grams per
minute with a bacteria/stearic acid slurry of 35~
bacteria, 65~ stearic acid. When this is done, a
product having a particle si2e of from 75 microns to
300 microns will be achieved, with a preferred level
of less than 250 microns.
The following examples are offered to further
illustrAte, but not limit, the process of the present
invention~ The examples are described in connection
with Figures 1, 2 and 3. Examples 1 through 4 and
Figures 1, 2, and 3 relate to the invention of my
prior case. Example 5, and tables 2-10, relate to
the process of this present invention for a poultry
probiotic.
Example 1
Example 1 correlates with Figure 1~ It shows
th~ product stability of two different strains of
Enterococcus faecium with temperatures of 4C and
27C. As illustrated ln Flgure 1, it shows a
stability of the encapsulated strains of En~ero-
coccus faecium, with the encapsulation being by the
rotary disc device using stearic acid with a level of
35~ culture weight. Conditions of microsphere
ormation were as previously described herein, namely
a 35/65 bacteria stearic acid slurry at a temperature
of 60C, using a four inch rotary disc, operating at
,
WO93t0~08 PCT/~IS92/075~')
- 21~65~5
11
3000 rpm and a feed rate of 100 grams per minute.
The spheres were formed, placed in heat sealed vapor
barrier pouches and destructively sampled waekly fQr
CFU determination. It can ~e seen that the product
of the invention maintained ~xcellent organism colony
forming unit (CFU) counts out to storage times at
long as 70 days.
Exam~le 2
Example 2 is to be interpreted in connection
with Figure Z. The figure shows the stability of
individual microsphered strains when mixed in a
typical feed ration in the presence of three poultry
antibiotics. The ration consis~ed of the following:
54% five cracked corn
26~ soybean meal
2~ fish meal
1~5~ dicalcium phosphate
1~ limestone
5 . ~f% soy oil
' 12% moisture content
Three antibiotics were added at the following
inclusion rates by weight: deco~uinoate 6% (454
ppm), salinomycin (50 ppm) and monensin sodium (120
'! ppm)-
~f~ Culture was added to the mixture at a level to
deliver approximately lxlO6 CFU/gm feed. Feed was
packaged in heat sealed bags and incubated at room
-, temperature. Samples were taken weekly for CFU
~ determination. The graph of Figure 2 illustrates the
,
, excellent stabili~y.
,,,
}l :
, f,
`~:
:
WO93/0620B PCT/US92/0758')
Example 3
Example 3 is to be interpre~ed in conjunction
with Figure 3. It shows the stability of the . .
Enterococcus faecium microspheres in eed in the
presence of different antibiotics. The ration
consisted of 60~ fine cracked corn, 38~ soybean meal
and 2% limestone with a moisture content of about
14%. Culture was added to a level of approxima~ely
106 CFU/gm feed and mixed. ~en pound ali~uots were
stored in sealed bags at 20 C and sampled weekly for
16 weeks. The antibiotics were included in the
ration at the following levels:
Bacitracin methylene disalicylate .... 50 gm/ton
Carbadox ............................. 50 gm/ton
Chlortetracycline ................... 200 gm/ton
Lasalocid ............................ 30 gm/ton
Lincomycin .......................... 100 gm/ton
Neomycin ............................ 140 gm/ton
Oxytetracycline ..................... 150 gm/ton
Sulfamethazine ...................... 100 gm/ton
Tylosin ............................. 100 gm/ton
Virginiamycin ............-................ 20 gm/ton
ASP250 .....^............................. 100 g~/ton
Furadox ................................... 10 gm/ton
Table 1 is a list of the minimum times ~or a 1
` log l~ss in colony forming units (CFU).
f
3;
,
t~ .
W093/0620~ PCT/~;~92/075~')
~11 652a
- 13 -
Table 1
Tlme in weeks for loss of 1 log CFU counts
at 20C in 14% moisture mash feed.
Antibiotic Time of Storaqe (da~s)
Control 103
Bacitracin 88
Carbadox 54
Chlortetracycline 60
Lasalocid 57
Lincomycin 75
~eomycin 53
Oxytetracycline 59
Sulfametha~ine 62
Tylosin 52
Virginiamycin 112
: ASP250 : 67
' ~ Furadox 53
., ~
Example 4
. ~
In Example 4 the:s~ability of product after
pelletizi~g for ~se of a chicken feed product was
~: determined. The microsphere formation conditions
were as earlier described. The conditions used in
this study were tha~following: -
.
Crude Protein, not less than ....... ~..... 18.0%
. Crude Fat, not less than ................. . 5.0~
Crude Fiber, not more than .........'...... 6.0%
The pellets with~and without the antibiotic (CTC
50 gm/ton~ were made with the following ingredients
. and conditions.
Corn, SBM, whey, soy oil, dicalcium phosphate,
limestone, tr~Ge mineral premix, vitamin premix,
~ selenium, copper sulfate. Culture was added at
i~ approximately 5x105 CFUlgm feed.
,
.
', :~
W093/062~ PCT/US92/075X'J
,
- 2~ 2~ 14 -
Conditioning temperature was 70C and the
pellets out of the dye were 78C.
Pelle~s were stored in unsealed bags and sample~ .
weekly for CFU determination.
In each instance the pelletized product was not
adversely affected ln stability by the conditions of
pelletizing. In particular, the pelletized product
showed stability egual to the unpellstized product.
Example 5
: Four thousand five hundred sixty, day-old
Peterson x Arbor Acres broiler chicks were randomly
assigned to floor pens (Table 2) with reconditioned
litter and fed for 45 days. All birds dying during
the first 5 days were replaced with a same-sex bird
from the same shipment and same treatment. The
composition of the basal starter, grower, and
I withdrawal rations i5 shown in Table 3. Star~er,
.31 grower f and withdrawal rations were formulated to
j con~ain 1425, 1450, and I475 kcal ME/lb,
respectively, wi~h 90 g/ton monesin. Starter rations
~ were fed from l to 21 days of age, grower from 21 to
`~ : 42 da~s Qf age, and wi~hdrawal from 42 to 49 days of
i ag~. The treatments were negative control, mash
~ (Contrbl, M); a:selected, encapsulated probiotic
`; cultures:~containing EnterocQccus faecium 301, DSM No.
D5M-Nr. 4789 and Enterococcus faecium 202, DSM No.
DSM-Nr. 4788 each rotary disc fatty acid encapsulated
l as described in Example 1 and each present as 50% of
'. the probivtic applied at 1 x 10 CFU/g of feed, mash
(prob~otic, M); negative control, pelleted (Control,
~jJ~, P); probiotic applied at 1 x 106 CFU/g mash, pelleted
` (probiotic, P) and a positive control applied at 10
`: g/ton virginiamycin, pelleted (Stafac ~ 10). The
. starter ration was crumbled for the treatments that
i.~ were pelle~ed. Twelve replicated pens of 35 males
.,
,, ,
. ~
~093/~6~0X PCT/U~92/07~X"
2116525
- 15 -
and 35 females were used with each experimental
ration.
Body weights, feed consumption, and mortality.
after the first 5 days were recorded by pen. Feed
conversion, adjusted feed conversion, and body-weight
adjusted feed conversion were calculat~d ~or each
pen .
All data were sub;ected to analysis variance and
differences were d~termined using Fisher LSD.
Prior to the study, probiotic culture
concentrate was extended with calcium carbonate. The
theoretical counts for probiotic, M and probiotic, P
were 1 x 108 and 2 x 109 CFU/g of product,
respectively. An 11 g sample of each product was
assayed in duplicate to determine actual product
counts. Each sample was plated using the Pioneer
standard plating technique~for encapsulated lactic
acid bacteria.
A mixer test was conducted for each production
phase, The test was designed to ensure that the
probiotic was uniformly distributed at appropriate
levels in the feed and~hat it sur~ived pelleting.
Each ~atch was sampled at the time of bagging with 4
equally spaced samples for the mash treatments and 10
equally spaced samples for the pelleted treatments
e. bags 1, 3, 5,..., 35, 37, and 39).
Alternate 1Oor pens within a treatment had non-
contaminated feed sampled during weeks 1 and 4; with
the remaining pens sampled on weeks 2 and 6 during
the feeding study.
An equaI number of birds from each sex was
sacrificed for the determination of individual
~ ~ breast, body and small intestinal weights, and small
!i intestinal length. Breast yield and intestinal
weight and length ratios were calculated for each
bird.
;,
~:
;'i
W093/0620X PCTi~lSg~/07~"
2116525
- 16 -
All data were subjected to a split-plot analysis
o~ variance and differences were determined using
contrast and estimate statements for the desired . .
e~fects.
Sixty birds per treatment were transported to a
university for a sensory taste panel evaluation.
Probiotic, regardless of processing, improved
(P<.05) feed conversion over the respective Control
while increasin~ (P~.05) weight gain over the Control
only in the mash feed (Table 4). The probiotic, P
improved tP~.05) feed conversion over Stafac ~ 10
which was similar (P>.05) to Control, P.
The product was at its desired level and strair;
composition (Table 5).
Probiotic was uniformly distributed within the
feed. Probiotic, M was at its desired level while
probiotic, P was 1 to 1-1/2 log higher than desired
for the starter and grower rations (Table 6). The
high counts for probiotic, P were a result of
overengineering of the product to ensure sufficient
recovery of the organisms af~er pelleting.
The floor pen samples for the probiotic, P
corresponded closely with the counts from the mixer
tests ~Table:7). However, probiotic, M dropped 2
logs in weeks 4 and 6 in the grower and withdrawal
mixes.
Probiotic, M increased (P<.05) both breast
weight and yield over the Control, M (Ta~le 8) while
probiotic, P showed an improvement (P>.05) over
Control, P. The improvement in the mash feed agrees
with the results found in an earlier trial. The
probiotic, P did not show a similar magnitude in
improvement in breast yield to that observed in
probiotic, M. This failure may be due to improved
ener~y utilization by pelleting resulting in less
room for improvement.
;,
-i
WV93/06208 PCT/~iS92/~75~')
2116~2~
- 17 -
Pelleting increased the average bird weight by
~6 g over mash. Pro~iotic increased the uniformity
of bird weights (Figure 5) with the greatest
improvement is mash feed.
Pelleting increased the average breast weight by
15 g over mash. Probiotic increased the average
breast weight and uniformity (Figure 4) over the
Control with the greatest improvement f ound in mash .
Stafac ~ 10 showed the greatest improvement in
uniformity for the pelle~ed feeds.
Pelleting increased breast yeild by .53
percentage units over mash. Probiotic, M showed a
.84 percentage unit incxease over Control, M which
was similar in magnitude to the pelleting respoonse.
The probiotic treatments produced a shorter
( P> . 05 ) small intestinal length than either of the
Controls and Stafac ~ when expressed as actual
length, a ratio of either body weight, or breast
weight (Table 9). Probiotic, M had a lighter (P>.05)
small intestinal weight than Con~rol, M when
~expressed as either actual weight or percentage of
A~ either body or breast weights. The reduction in
intestinal weight and length for probiotic treatments
suggests less energy required for maintenance and
more energy available for growth as indicated by
improved feed conversion and breast yield (Table
7-8).
The probiotic, P treated birds produced no off-
flavor when compared ~o Stafac ~ 10 (Table 10). In
~, ~ the second trial, probiotic, P was perceived to have
enh~nced the flavor of the thigh/leg when compared to
Control, P. However, this enhancement of flavor was
not observed in the first trial.
' ?~
C
.?, ' , , ' ' ~ , ,' i~ ' i
''"i
!
-1 8 ~ 2 !~
TABLE 2
PEN ASSIGMMENTS
Treatments Pen numbers
-
Control, P 2,6,15,17,22,26,104,10g,113,117,122,1Z6
Probiotic, P 4,8,12,16,21,28,105,106,112,118,125,130
Stafac0 10 5,7,11,18,23,27,101,107,111,116,123,129
Control, M 3,9,13,20,24,30,102,10~,114,11g,121,127
Probioti~, M 1 , 10 , 1 4 , 19 , 25 , 29 , 103 , 110 , 115 , 120 , 124 , 128
Pen size 4.2~ x 15.5~, one tube feeder, one hanglng wate~er, pine
shavings on dirt/ power and evaporative eooling system and well
insulated, forced hot-air heat, curtain sidewall building.
TABLE 3
COMPOSITION OF BASAL R~TIONS
i
Production Phases
3 Ingredients Starter Grower ~ Withidrawal
l~ ~ Ground corn ~ 65.37 67.89 74.29
i So~bean meal 25.58 23.53 17.83
~ ~ MFett and bone ~ieal ~ 3 6 3 32 3 00
~ Deflourinated p~ospha~e ~95 .79 .73
Calciu~ carbonate ~ 615 631 32
Trace m~neral .05 .05 .05
Methionine 19 06 18
Vitamin premix .05 .05
Y
..
.,: .
. i .
'I : . ~;
~,;j ..
, ,
-1~9-
21~6S25
TABL~ 4
FLOOR PN PRODUCTION DA~
PolletStafac~ lnv ~
Control P 10 Control M
_
~eight, lb. 4.79~ 4.81~4 79~ 4,54b 4.6~-
~eed conv. l.B71b 1.827' l.B55~ b 1 . 917c 1,~56
Adj. feed conv.l 1.832b 1.789~07~ 1.8~7C 1.512
We ght, adj 2 l.B01b 1.755' 1.775 1.897C 1.798b
Mortality, ~ 4.40 4.64 5.95 3.33 5.60
Adj feed conversion ~ Total feed/~live + dead weight).
Weiaht adj. feed conversion Y Adj. feed conversion~ weight-4.60)/6).
~b~ p< 05
i' * Inv = Invencion
TABLE 5
PRO W CT QC AND QA
Treatments ~ count ~A2count Strain ratio
1:
l Probiotic, P S.75~x 10~ 1.01 x 10~ 50:50
Probiotic,~M , : :~.54 x 107 9.60 x~10 57:43
, :
1. Qualit~ control
. Quality assurance.
~'s
~i: :
;~ :
:
,t ~ :
:~ :
. ', .
-20- 2116~5
TABLE 6
FEEDMILL MIXER IEST AND RECOVERY
Produc~tion Phases
and TreatmentsMash Pellet . Recover~
~ ~ cfu/g of ~eed - % ma~;~ ~~~
Probiotic. P 2.02 x ~o6 1 67 x 10~ 98.69
Stafac~ 10 NA 6.46 x 10
Control, M Z.51 x 103
Probiotic, M 1.34 x 10
Grower
Control, P NA 4.86 x 102
Probiotic, P 3. as x 106 1 . 09 X 106 91 . 62
Stafac~ 10 5.25 x 10~ 6.42 x 103
Control, M 1.00 x 102
Probiotic, M 1.48 ~ 10
Probiotic, p B 50~x 10~ 1 11 x 103 117.40
Stafac~ 10 8.80 x }03 1.79 x 10
Control, M 8.92 x 10
Probiotic, M : 1.33 x 10
Mean
Control, P : 8 50 x 102 8.28 x 102
Probiotic, P 8 2~ x 1~5 9.64 x lQ6 llB.09
Stafac~ 10 2.15 x 104 9.05 x 10 -
Control, M B.72 x 102
~: Probio~ic, M ~ 1.38 x 105
Recovery calculated on logiD transformed data.
2 NA means not available.
.
::;
-21- 2116525
TABLE 7
EIOOR PEN Q~
.
Weeks
Treatment:s 1 2 4 6 Mean
- cfu/g of leea
Control 3.78 x 102 3.83 x 1028.60 x 102 2.21 x 102 4.08 x 102
Probiotic, P9.23 x 1059.37 x 1058~77 x 105 8.4~ x 105 8.96 x 105
Stafac~ 10 8.73 x 102 1.29 a~ 1026.46 x 102 8.63 x 102 8.89 x 10
Control, M 3 46 x 102 1.26 x 1022.79 x 103 2.00 x 102 5.08 x 10
Probiotic, M 1 43 x 1051.25 x 1051.75 x 103 1.00 x 103 2.32 x 104
.
TABLE 8
BRE:AST YIELD EVALW~TION
PIel * -- stafac~ Inv *
ontrol P 10 Control M
Body weight, g ~ ~ ~ 2240 7 2230 1 2195 9 2143 8 2149 9
~ ~ Br~ yield, % of ; ~ 10 . 68A 10 . 58~ 9, 93b 10 . 67A
?i ~ a b P< 05
* Inv = Invention
! :
,,;
:~
,
^.';1
i';
.:~:
/
2116~
TABLE ~
IN~STINAL t~EIGH~ AND LENGTH
Pelle~t Stafac~
Control P 10 Control
_ . _
Body weight, g 2240.7 2230.1 2195.9 2143.~2149.9
Breast weight, g 234.4~ 239.6~ 232.0~ 213.3b 229.6'
SI weight~ g 92.6 3.3 93.4 91.4 .87.4
SI, g/in 71 31 715 23 71 22 71 107~ 16
SI we ght~ g/100 g4.17 4.18 4.27 4.29 4.08
SI length, in/loo g
body weight 3.47 3.40 3.53 3.61 3.53
SIbreagt weight 40.19 39.70 40.97 43.96 38.69
SI length, ig~lt 33.41~ 32.27~ 33.72~ 36.89b 33.41-
`!
J, ab P~ . 05
3; * Inv = Invention
~ ~ SX ~ Small Inte~ine
i~' - TABLE lO
~ ~ TASTE PAN~ EV~IL~TION
s
- Group ~ - umbIr of correct identifications
Tissue Co~parison : Trla Tria 2 C ne
gh,/leg Stafac~:10 YS. Co~trol, P 6 3 9
Stafac0 lC vs. XINOC, P 3 4 7
- Probiotic, P vs. Control, P 2 8* 10
: Breast: Stafac~D lO vs. Controlr P 2 6 8
- ~ Staac0 lO v~. XINOC, P 1 3 4
P~obiotic, P vs~. Control, P 5 4 9
The evaluators were able to detect the odd sample a statistically
signlficant (P<.05) n~ber of times.
he number of correct identifications o~ the Oda siæmple required for
significance at the 5~ level was 7 for n=10 and 11 for n=20.
~q ~
.. ~
l,,j
,. ~ .
. ,, , .. . , . . ~ .. .. . . . .. ~
