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 Materials and methods
 Experimental
 Results and discussion
 Summary
 Literature cited














Group Title: Animal husbandry and nutrition mimeograph series - University of Florida Animal Husbandry and Nutrition Dept. ; no. 61-4
Title: The Effect of ante-mortem injection of papain on tenderness of chickens
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Full Citation
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Permanent Link: http://ufdc.ufl.edu/UF00072903/00001
 Material Information
Title: The Effect of ante-mortem injection of papain on tenderness of chickens
Physical Description: 7 leaves : ; 28 cm.
Language: English
Creator: Huffman, D. L., 1931-
Palmer, A. Z.
Carpenter, J. W.
Shirley, R. L.
Publisher: Florida Agricultural Experiment Station
Place of Publication: Gainesville, Fla.
Publication Date: 1961
 Subjects
Subject: Chickens -- Marketing -- Florida   ( lcsh )
Chickens -- Quality -- Florida   ( lcsh )
Papain -- Research -- Florida   ( lcsh )
Genre: bibliography   ( marcgt )
government publication (state, provincial, terriorial, dependent)   ( marcgt )
non-fiction   ( marcgt )
 Notes
Bibliography: Includes bibliographical references (leaf 7).
General Note: Caption title.
General Note: "January, 1961."
Funding: Animal husbandry and nutrition mimeograph series - University of Florida Animal Husbandry and Nutrition Dept. ; no. 61-4
Statement of Responsibility: D.L. Huffman ... et al..
 Record Information
Bibliographic ID: UF00072903
Volume ID: VID00001
Source Institution: University of Florida
Rights Management: All rights reserved by the source institution and holding location.
Resource Identifier: oclc - 77068673

Table of Contents
    Materials and methods
        Page 1
    Experimental
        Page 2
    Results and discussion
        Page 3
        Page 4
        Page 5
    Summary
        Page 6
    Literature cited
        Page 7
Full Text





Animal Husbandry and Nutrition Florida Agricultural
Mimeograph Series No. 61-4 Experiment Station
January, 1961 Gainesvilie, Florida


THE EFFECT OF ANTE-MORTEM INJECTION OF PAPAIN
ON TENDERNESS OF CHICKENS

D. L. Huffman, A. Z. Palmer, J. W. Carpenter and R. L. Shirley /


The effect of the administration of papain by ante-mortem injection
in meat animals has been investigated by Beuk et al. (2) and Palmer et al.
(4). According to Beuk et al., a tenderizing effect resulted when papain was
injected ante-mortem into chickens, sheep, goats and cattle; a shorter ante-
mortem time was required for tenderization when the papain was injected into
the blood stream than when the injection was made into the peritoneal cavity.
Ante-mortem treatments were administered from two minutes to two hours before
slaughter for an intravenous injection and from six to 24 hours for an intra-
peritoneal injection.

Thomas (5) reported that a more purified crystalline papain did not
produce the same histological changes as a crude papain preparation when
injected in rabbits. Thus, the report introduced the possible additive
effect of an enzyme activator present in the crude preparation but not in
the crystalline preparation.

The purpose of this study was to compare the tenderizing effects of
crude and crystalline papain preparations using chickens as the test animal.
A further objective was to determine whether or not an enzyme activator
was present in the crude papain and if present, whether or not the factor
was effective as a tenderizing agent alone or only when injected with the
enzyme.

Materials and Methods

The crystalline papain used in this study was prepared by the method
of Balls and Lineweaver (I). The activity of the crystalline and crude
papain preparations was assayed by the method described by Weiner (6).
The assay indicated that the crystalline preparation was approximately
100 times as active in vitro as the crude papain preparation on a dry
matter basis. The crystalline preparation was in a 1:100 dilution uEN
a O.IM cysteine solution.

The chickens used in each experiment were of uniform sex/ e and oC i
breeding. Since sex, age and breeding differences exist betw exp erjnrts
the data from one trial should not be compared directly with r o r
experiment. The injection technique followed was similar to used \

/ Huffman, Research Asst.; Palmer, Associate Animal Husbandman;'.. '
Carpenter, Assistant Animal Husbandman and Shirley, Animal Husbandman,
Department of Animal Husbandry and Nutrition, University of Florida,
Gainesville, Florida. The assistance of Mrs. Grace B. Howes and
Mrs. Norma Bowles is acknowledged.









by Palmer et al. (4). The chicken was held on it's back and the solution
injected into the peritoneal cavity at a point midway between the posterior
end of the keel bone and the vent. A I1 inch, size 18 needle and an
ordinary glass syringe were used to inject the solution.

Crude papain solutions were mixed so each ml. of solution contained
10 mg. of papain or the equivalent amount of crude papain ash. The crude
papain was ashed at 500 C. for 12 hours in a muffle furnace. The ash was
then washed into a flask, distilled water was added to make up the desired
volume and the mixture agitated until a uniform suspension was observed.
The more concentrated solutions of ash were prepared similarly except that
each ml. of distilled water contained ash from 20 and 40 mgs. of crude
papain, respectively.

The crystalline and ash solutions were prepared by using the solution
of ash and distilled water and adding the crystalline papain at the rate of
0.1 mg. per ml. of water. The crude papain was denatured by first diluting
to the desired concentration with distilled water and heating in a water
bath at 970 C. for 30 min.

The chickens were weighed to the nearest gram and dosages calculated
on the basis of body weight. The chickens were not fed or watered during
the holding time of six and 12 hours. At the end of the holding time rectal
temperatures were taken and the chickens slaughtered, dressed and drawn.
The carcasses were weighed, placed immediately into a slush ice bath for 24
hours, reweighed and either cooked immediately for taste panel evaluation or
frozen for later evaluations. The chickens were placed breast side up on
roasting pans in electric ovens set at 3500 F. and roasted at the rate of
one minute per 12 grams of dressed weight. Small birds were roasted a
minimum of one hour to obtain a degree of doneness comparable to larger
birds. A trained taste panel consisting of four members evaluated each
sample on a tenderness scale of one to nine with one being too tough to
be edible, five being average, and nine being over tender or mushy.
Samples were taken for taste panel evaluation from the breast muscle
(Pectoral maior).

Experimental
in Trial I, thirty white Leghorn roosters, approximately eight months
old, were randomly divided into six groups of 5 birds each for treatment.
Treatment I birds were designated as the injected control and the birds
received 3.5 mi. distilled water per kg. body weight. Treatment 2 was an
injected control and the birds received 5.5 ml. distilled water per kg. body
weight. Treatments 3 and 4 received 0.35 and 0.55 ppm crystalline papain,
respectively. Treatments 5 and 6 received 35 and 55 ppm crude papain.
Injections, based on live weight, were administered six hours ante-mortem.

In Trial 2, twenty-four white Leghorn roosters approximately eight
months old, were randomly divided into six treatments of 4 birds each.
Treatments I and 2 were the injected control groups and received 5.0 and
10.0 mi. distilled water per kg. body.weight, respectively. Treatments
3 and 4 received 0.5 and 1,0 ppm crystalline papain, respectively.
Treatments 5 and 6 received 50 and 100 ppm crude papain, respectively.
Injections, based on live weights, were administered 12 hours ante-mortem.


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In Trial 3, sixty where Leghorn hens, approximately two years old,
were randomly divided into six groups of 10 birds each. Treatment I
was the non-injected control group. Treatment 2 was I ppm crystalline
papain. Treatment 3 birds received 100 ppm crude papain. Treatment 4
was 100 ppm denatured crude papain. Treatment 5 birds received ash from
crude papain, in the amount ashed from 100 ppm crude papain in distilled
water. Treatment 6 was I ppm crystalline papain plus ash in the amount
ashed from 100 ppm crude papain. Injections, based on live weight, were
administered six hours ante-mortem.

In Trial 4, thirty-five white Leghorn hens, approximately 2 years old,
were randomly allotted to seven treatments. Treatment I was the non-
injected control group. Treatment 2 received 10.0 ml, distilled water per
kg. body weight containing ash in the amount ashed from 200 ppm crude papain.
Treatment 3 received 10.0 ml. distilled water per kg. body weight containing
ash in the amount ashed from 400 ppm crude papain. Treatments 4, 5 and 6
received ash in the amount ashed from 100, 200 and 400 ppm crude papain
respectively along with I ppm crystalline papain. Treatment 7 received 100
ppm crude papain. Injections, based on live weight, were administered six
hours ante-mortem.

Results and Discussion

In Trial I, neither the crystalline papain nor the crude papain
significantly influenced the tenderness of breast muscle. Lack of sig-
nificance might be attributed to dosage levels in view of later trials of
this study. Trial I data are presented in Table I.

TABLE I. THE EFFECT OF CRYSTALLINE AND CRUDE PAPAIN
ON THE TENDERNESS OF BREAST MUSCLE (PECTORAL MAJOR). -- TRIAL I.


Treatment ml. of Dist. Papain No. Av. Tenderness-/
Water Per kg. ppm of Body Wt. Birds Breast Muscle
Body Weight.

I. Injected control 3.5 -5 5.55

2. Injected control 5.5 5 6.65

3. Crystalline papain 3.5 0.35 5 5.75

4. Crystalline papain 5.5 0.55 5 6.20

5. Crude papain 3.5 35 5 6.30

6. Crude papain 5.5 55 5 6.65

I/ Differences were not significant.


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In Trial 2 the levels of crystalline and crude papain injected were
increased over the levels used in Trial I. Even though the number of birds
per treatment was only 4 in the trial, the injection of 100 ppm crude
papain increased (P(O.05) the tenderness of breast muscle compared to
all other treatments. Other treatments had no significant tenderizing
affect as compared to the injected control groups. Trial 2 data are presented
in Table 2.

TABLE 2. THE EFFECT OF CRYSTALLINE AND CRUDE PAPAIN
ON THE TENDERNESS OF BREAST MUSCLE (PECTORAL MAJOR). -- TRIAL 2


Treatment ml. of Dist. Papain No. Av. Tenderness
Water Per kg. ppm of Body Wt. Birds Breast Muscle
Body Weight

1. Injected control 5 -4 6.81

2. Injected control 10 -4 6.75

3. Crystalline papain 5 0.5 4 6.25

4. Crystalline papain 10 I 4 6.44

5. Crude papain 5 50 4 6.69

6. Crude papain 10 100 4 8.25 -/

1/ Differed significantly (P<0.05) from all other treatments.

Trial 3 data are presented in Table 3. It is interesting to note
that all injected treatments were more tender than the non-injected
control birds according to average tenderness values. However, only
treatments 3 and 6 were significantly more tender than the controls.
Treatment 3, the 100 ppm crude papain injection, was more tender than
Treatments I, 2, 4, 5 and 6 with differences being highly significant
(P<0.01). The average tenderness value for Treatment 3 birds indicates
over tenderization sufficient to cause a lack of palatability; the meat lacked
in body, texture and chewiness. Treatment 6, I ppm crystalline papain plus
ash of crude papain, was more tender than Treatment I (P<0.01) and Treatment
5 (PO0.05). It should be noted that Treatment 2, I ppm crystalline papain,
and Treatment 5, ash of crude papain, were more tender than Treatment I,
the control group, according to average values. Treatments 2 and 5 were
combined into single Treatment 6, I ppm crystalline papain plus ash of crude
papain, and the increased tenderness resulting, indicates a combining,
synergistic and activating effect.


-4-









TABLE 3.


THE EFFECT OF CRYSTALLINE PAPAIN, CRUDE PAPAIN AND
ASH OF CRUDE PAPAIN ON THE TENDERNESS OF BREAST
MUSCLE (PECTORAL MAJOR). -- TRIAL 3.


Treatment ml. of Dist No. Av. Tenderness
Water/Kg Birds Breast Muscle
Body Wt.


I. Non-injected control 10 6.08

2. Crystalline papain I ppm of body wt. 10 10 6.63

3. Crude papain 100 ppm of body wt. 10 10 8.25-

4. Denatured crude papain 100 ppm body wt. 10 10 6.50

5. Ash of crude papain (equiv. to ash
in 100 ppm crude papain of body wt.) 10 10 6.35

6. Crystalline papain, I ppm of body wt.
plus ash of crude papain (equiv.
to ash in 100 ppm crude papain
of body wt.) 10 10 7.03 2/

/ More tender than I, 2, 4, 5 and 6 (P<0.01)
2/ More tender than I (P<0.01) and 5 (P(0.05)


A further study of the tenderizing effects of crude papain ash,
crystalline papain plus varying levels of crude papain ash, and crude
papain is summarized in Table 4. In Trial 4, as in Trial 3, all in-
jected treatments were more tender than the non-injected control group
according to average-tenderness values. Treatment 2 was more tender
than control Treatment I (PO0.05). Treatment 4, I ppm crystalline papain
plus ash of crude papain, was more tender than Treatment I (P<0.01), and
Treatment 3 (P Treatment I (P(0.01). Treatment 7, 100 ppm crude papain, was appreciably
more tender than all other treatments (P'0.01). Treatment 7 birds were
over-tenderized to the point of lacking acceptability. Findings of
Trial 4 confirmed the findings reported for Trial 3 notably that ash of crude
papain exerted a notable tenderizing effect, that 100 ppm crude papain
significantly tenderized and that I ppm crystalline papain and ash of crude
papain when combined gave tenderization indicating a complimentary effect.


-5-









TABLE 4. THE EFFECT OF CRUDE PAPAIN ASH, CRYSTALLINE PAPAIN
PLUS CRUDE PAPAIN ASH AND CRUDE PAPAIN ON THE
TENDERNESS OF BREAST MUSCLE (PECTORAL MAJOR). -- TRIAL 4


Treatment ml. of Dist No. Av. Tenderness
Water/kg. Birds Breast Muscle
Body Wt.


I. Non-injected control 5 4.60

2. Ash of crude papain (equiv. to ash
in 200 ppm crude papain of body wt.) 10 5 5.75

3. Ash of crude papain (equiv.to ash
in 400 ppm crude papain of body wt.) 10 5 5.40

4. Crystalline papain, I ppm of body wt.
plus ash of crude papain (equiv.to
ash in 100 ppm crude papain of cr
body wt.) 10 5 6.20 /

5. Crystalline papain, I ppm of body wt.,
plus ash of crude papain (equiv. to
ash in 200 ppm crude papain of body 3/
wt.) 10 5 6.05

6. Crystalline papain, I ppm of body wt.,
plus ash of crude papain (equiv. to ash
in 400 ppm crude papain of body wt. 10 5 6.05

7. Crude papain 100 ppm of body wt. 10 5 8.40 -

I/ More tender than I (PO0.05)
/ More tender than I (P 3/ More tender than 3 (P<0.05)
4/ More tender than I (P I/ More tender than all other treatments (P<0.O1)


Four trials, involving 149 white Leghorn young roosters and mature
hens, were conducted to compare the tenderizing effects of crude and
crystalline papain preparations injected into the peritoneal cavity and
to determine the presence and activity of an enzyme activator or synergist
in crude papain.


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Ante-mortem injection of 0.35 and 0.55 ppm crystalline papain
had no significant effect on the tenderness of breast muscle.

Ante-mortem injection of 35 and 55 ppm crude papain had no signifi-
cant effect on breast muscle.

In three trials, 100 ppm crude papain injected ante-mortem tenderized
breast muscle either at significant or highly significant levels of probability.
Over tenderization, however, rendered the breast muscle lacking in sufficient
body or texture to be fully acceptable.

Findings of Trial 4 confirmed Trial 3 data in that ash of crude
papain exerted a tenderizing effect and that crystalline papain and ash
of crude papain when combined gave tenderization indicating a compli-
mentary effect.


Literature Cited

Balls, A. K. and H. Lineweaver. 1939. Isolation and properties of
crystalline papain. Journal of Biological Chemistry, 130:669.

Beuk, Jack F., Alfred L. Savich and Paul A. Goeser. 1959. Method of
tendering meat. United States Patent 2,903,362. Patent issued
September 8, 1959.

Duncan, D. B. 1955. Multiple Range and Multiple F Tests. Biometrics,
11:1-42.

Palmer, A. Z., R. H. Alsmeyer and J. W. Carpenter. 1957. Effect of
ante-mortem injection of papain on the tenderness of mature
chickens and beef. Annual Report, Florida Agricultural Experi-
ment Station.

Snedecor, G. W. 1956. Statistical Methods. 5th Edition. Iowa State
College Press, Ames, Iowa.

Thomas, Lewis. 1956. Reversible collapse of rabbit ears after intra-
venous papain and prevention of recovery by cortisone. Journal
of Experimental Medicine, 104:245.

Weiner, S. 1956. Papain, a review of the literature. Paul Lewis Labora-
tory publication.







Mimeo Series 7 -
DLH:rw
1/18/61




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