Of Cured Pork Legs by M
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[ 165 ] INTERNAL BACTERIAL TAINTS ('BONE TAINT' OR 'SOURING') OF CURED PORK LEGS BY M. INGRAM Low Temperature Station for Research in Biochemistry and Biophysics, University of Cambridge, and Department of Scientific and Industrial Research (1) INTRODUCTION The bacteriology of ham souring has been comprehensively reviewed, in relation to American packing-house practice, by Jensen (1944, 1945). Observations accu- mulated over many years at the Low Temperature Station suggest that Jensen's conclusions may not be wholly applicable in Britain and, though covering an amount of material much smaller than that available to the American workers, they suffice to suggest some new points of view. There is, in fact, still very little published information about the bacteria normally found in such products, a situation recently deplored both by Mundt & Kitchen (1951) and by Ulrich & Halvorson (1951). Moreover, in the past few years, British work has revealed influences (reviewed briefly by Ingram, 1948) which Jensen did not consider. Hence it seemed worth while to give a connected account of our observations in this paper, which surveys the species we have isolated. A distinction will be made between 'bacon gammons' and 'hams': the former term being used for legs of pork into which considerable proportions (5 % or more by weight) of brine have been injected during curing (by pumping through a long hollow needle); the latter term denoting similar material which has been injected only lightly, or not at all. It is characteristic of present-day tank-cured bacon that it is always thoroughly injected with brine to hasten the cure; whereas in the traditional British cure of hams, still used in home-curing (Ministry of Agriculture and Fisheries, 1945) and approached in some commercial cures, salts were applied only to the outside and nothing was injected into the interior, and the same procedure is still used in the Southern 'country-style' hams of the U.S.A. (Mundt & Kitchen, 1951). Distinction on this basis is admittedly rather arbitrary, in that some hams are nowadays injected while an inefficiently injected 'gammon' would approach the state of a 'ham', and there are additional minor differences between the curing of hams and that of gammons. Nevertheless, the choice of terms has a reasonable basis, and it will be useful in emphasizing the point of difference which is fundamental to the present theme. (2) METHODS The problems of obtaining deep bacteriological samples from leg joints, without introducing unwanted contamination from surface organisms, were discussed by Haines (1941), and similar procedures were used in this work. A flat surface for searing was prepared, by cutting through the leg with roughly sterile instruments, after the gross bacterial load on the external surfaces had been removed by washing 166 M. INGRAM and disinfection with Lysol. The cut passed perpendicular to the femur, just above the stifle-joint, so as to expose the marrow of the femur and the head of the gastrocnemius muscle, from which 5 g. samples were dissected aseptically, after searing. This muscle was chosen because it has a relatively high pH (Table 1), and it always becomes involved when tainting of the flesh occurs. Table 1. pH of several different muscles in sound hams Ham Gastrocnemius Semitendinosus Sartorius Gracilis 1 6 16 6*01 5-92 5-58 2 6'16 5*95 6418 5.85 3 6-64 6-33 6*50 6*26 4 6*23 6X23 6-20 5X85 5 6-06 6X33 6X02 5X75 6 6.13 6-02 5*99 5.77 Average 6.23 6*15 6-13 5-84 After removal of the bacteriological sample, further portions of the muscle were taken for the following analyses: (i) The salt content was determined by electrometric titration of the chloride in a hot aqueous extract (Ingram & Hawthorne, 1945); but, as the effective factor is the concentration of NaCl in aqueous solution in the meat (Oxh0j, 1943; Hankins, Sulzbacher, Kauffman & Mayo, 1950), it is expressed as the percentage of the water content. (ii) The water content of meat was determined by drying a minced sample for 96 hr. at 1030 C. (values ranged from 65 to 75 % of fresh weight). (iii) The pH was found by snipping about 10 g. of muscle into 20 c.c. water, adding 0 5 ml. of CHC13, allowing to stand more than 2 hr. and measuring with a glass electrode at 150 C.: quantities and times were not critical. We have found (with psoas major muscles) that the gross pH of such samples is practically the same for different points in the muscle. (3) OBSERVATIONS (a) The flora of hams Sound hams are apparently nearly sterile internally. Examination of ten examples failed to yield any viable bacteria either in the bone marrow or the muscle; after which it seemed unnecessary to continue. In a tainted ham, in marked contrast, the spoiled regions contain enormous numbers of bacteria. Examination has shown that the typical flora consists of Clostridia and/or faecal Streptococci. Frequently this may be indicated simply by making a direct smear of the spoiled flesh and Gram staining, which usually reveals large Gram-positive rods and/or large ovoid Gram-positive diplococci. Gram- negative rods may be present, but they rarely predominate. Typical results of such direct examinations are given in Table 2. (Compare also Jensen, 1951.) The types of bacteria isolated from tainted hams when examined, on various Internal bacterial taints of cured pork legs 167 occasions and from different sources, are listed in Table 3.* As a group, these bacteria possessed the following characters: (a) They were predominantly mesophilic and did not grow well at low tempera- tures. More than 80 % of them had an optimum temperature of 370 C. (b) They were highly susceptible to salt. Less than 10 % of the isolates with- stood 10 % NaCl, and nearly 50 % of them (though growing well at lower concen- trations) failed to tolerate even 5 % NaCl in peptone water. (c) They were unable to reduce nitrate. Several of the species indeed, can be inhibited readily by nitrate (cf. Jensen, 1945, p. 14 et seq.). (d) They were unfavourably affected by moderately acid conditions (pH 5-5). This topic has been discussed elsewhere (Ingram, 1948). (e) There was a high prQportion of probably faecal types. Nearly half the isolations were plainly in this category and, of the remainder, a considerable number might have been so. Table 2. Approximate numbers of bacteria.per g. in tainted ham, estimated from direct microscopic examination Gram-positive cocci Gram-positive rods ,A __ Example A , Ovoid Strepto- Micro- Gram-nega- Miscel- no. Spores No spores diplococci cocci cocci tive rods laneous 1 108 - 107 - - 105 2 - - 107 108 - - _ 3 - - 106 10 - - - 4 - - 106 - - - 5 - - - 101 - 106 6 C 108 . 108 - _ + _ -. 106 107 7 - - 106 - 107 - Yeast 107 8 - 108 108 _ - _ 9 { - 107 108 + - + - X- lo's 10 - -1107 L lo6 106 Accordingly, one would expect tainting by the above bacteria to develop only where there is a relatively low concentration of curing salts, and where the muscles are less acid. The data included in Table 3 appear to confirm this in several cases. Unfortunately, however, the exact significance of such data is open to doubt. The salt concentrations are difficult to interpret because in some cases the ham was not available for some weeks after curing, by which time there had been ample opportunity for equalization of the salt concentrations by diffusion from the more readily penetrable portions of the leg. The values quoted in Table 3 may thus be considerably higher than those obtaining during the critical initial stages of the cure, as indeed seems very probable in a case like ham B 2, where three organisms * In this paper, the names are mostly those given in the 5th edition of Bergey's Manual of Determinative Bacteriology for species with the characters of the organism in question. In the later (6th) edition some of the groups, e.g. the micrococci, have been over-simplified for the purposes of this discussion. 168 . M. INGRAM 0 0 I I I I I I I I I I I I I + 0 4) 410 ce ++++ +I I +++ I I + I I I ++ I + w 0 4 ° 0 O2 g 14 V CO CO rO eq CO rO CO CO Cr eq rO O eq rO CO CO CO CO CO CO 0 X c cecq X cqsc oc sa ec oa ac ocrC :c C) Oq w4 Co oce 4 0* - 0 0 0 m Pai 0w . C.) 0 -9 .0 C~~~~~~~~~~~~~~~~I ab .5 P bo - m3 I(1)OD ~ ~ Co to P- co 0bo i--b oo * o X o C) . - eq O0 - CO CO cq 0 '0 1 CO? eD co cor Q C- EH 0 z _ eq co II - e) 0 ¢ 0 CO Internal bacterial taints of cured pork legs 169 +II+II+I++++++I+ I +++I+II+++I++I t- - r- r- 10 r 14 r- - r- - t- to t- - r- - r- - t- - r- - t- - r- - r- - r- t- ceXe o q e q c X ccqa csc ee ec c mc eXC o oc oc sc C) C). Zs~~~~~~~~~C *~ Z._4 * * .**4 . .¢ - 0.r -+'0) 4-.C. *C *XXtAga Satstl t e C) 49 00t *: t- *: . .£ . .- - t . ° z e . *s -&~ ** %-- IQ "' Zo b0), , ~ I "Z .Z t2 & ab CBo co 1b c CB c- Cl ct es4 cq m 1.0 e- q _ -I c9 _- aq co V C 4 PA -r J. Hygiene 12 170 M. INGRAM unable to resist even 5 % NaCl had grown in a muscle which contained 7-5 % NaCl when observed.