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Histamine in foods causing false positive scratch tests
Histamine in foods causing false positive scratch tests H. M. G. Doeglm, M.D., ad J. P. Xater, U.D., Rotterdam md The Hague, The Nethedntlds In. 80 normal test subjects, scratch trsts wcrc performed with a large nlrmber of wnextractcd foodstuffs. Urtiourial rcnctions wc’rc seen with certain cheeses, some brands of suuerkraut, and with yeast c~strrrc*ts. The reactions are cnusrd by the higlb histamine content of these foodstuffs.
F
ood hgpersensitivities have been of recurrent interest in the allergologieal literature. In the past, highly concentrated extracts or pure foodstuffs, which frequently caused irritant-type false positive reactions, were used for scratch tests. More recently, extracts have been used which, in many cases, were dialyzed to exclude small molecule irritant materials. The causes of these irritant false positive reactions usually have remained obscure. WC have not found any studies of the relationship of the possible histamine content of foods and the outcome of scratch tests. The presence of histamine in foods may be caused by different circumstances, such as: (1) histamine in the natural state of some foods. It has been demonstrated in: spinach, concentration 2.25 mg. per 100 Gm.,] tomato, no quantita’tive data,2 and meat, concentration 0.2 to 6.0 mg. per 100 Gm.3 (2) Histamine is formed during the production of foods by the action of bacteria. Histidine in these foods undergoes decarboxylation by the microorganisms. In this case, the ba.cteria take an active part in the production proces’s of these foods: cheese, concentration 0 to 100 mg. per 100 Gm.,4-6 sauerkraut, no qua,ntitative data,’ and wine, concentration 0 to 1 mg. per 100 ml8 No doubt, the influence of time is important, and histamine will occur more readily in foods which have to go through a period of ripening or aging. (3) Histamine may be formed in the food during spoilage. Bacteria with which the food has been contaminated may form From the Department Received for publication
164
of Dermatology, Sint April 17, 1968.
Franciscus
Gnsthuis,
Rotterdam.
histamine from histidine. influence and temperature MATERIAL
AND
In this case again, duration of exposure to bacterial may play a role, for example fish and meat.
METHODS
Scratch tests were performed on 20 healthy subjects. Only pure, whole foodstuffs were used. n-0 attempt was matle at extraction or dilution. Scratches were made with a vaccinostylc, taking cart to avoid bleeding. Solid foods were gently rubbed into the scratch. Hard materials, swh as old cheese, were heated slightly, until t,hey became soft and could be smeared into the scrat,ch. In each set of tests an “empty” scratch, to exclude dermographism, and a scratch with a solut,ion of 0.1 per cent histamine HCl was included as a, cont,rol test. The study was performed by 2 independent im-cstigators in different cities. In the same 20 subjects, scratch tests were done with a number of cheeses of different origin and age. This study was done blind; it was not known beforehand which cheeses might contain histamine. The histamine content of the cheeses was determined by the I)utch Institute for Dairy &search (NIX)), with a procedure developed by I’. J. dc Koning, Ph.D. ‘2 lo It consisted of high voltage elcctrophoresis of a. watery cheese extract on thin layers of silica gel, followed by fluorimetric analysis wit,11 o-phthalaldehytle. RESULTS Various foods
Negative scratch tests were seen in all persons rubbed with pure tomato, spinach, raw meat (beef), fish (sardines, haddock), and wine (Chianti, Gcwurtz-
Fig. 1 Scratch (1) blank sauerkraut (pure), dry matter.
control, (2) histamine HCI 0.1 per and (5) cheese (pure), containing
cent in mater, (3) 85 mg. histamine
tomato HCl
(pure), per 100
166
Doe&s
J. Allergy September, 1968
and Nater
traminer, Riesling-Sylvaner) . Positive urticarial reactions of diffcrcnt strengths were seen with the following foods (Fig. 1) : 1. Marmite, Yeastrel, and the salt-free variant,, Marrnex, are yeast extracts prepared by autolysis of yeast cells in a. salt solution. Thy contain up to 200 rng. of histamine ac,id phosphate per 100 (jm.ll 2. Histamine in sa.uerkraut, has been detcrmincd only qualitatively so far,’ in some, but not all, brands of sauerkraut. 3. Sardines, directly from a, freshly opened can, gave negat,ive reactions. However, after they had been kept, in an open plastic container at room tcmperature (18’ C.) for some weeks, the reactions became strongly positive. Ceigerl’ has established that the concentra.tion of histarninc, which is present only in traces in fresh sardines, incrcascs rapidly during spoilage up to values of 120 mg. per 100 (:m. of fresh meat within 24 hours. 4. The presence of histamine ha,s been demonstrated in yuantit,ies of 5 to 100 mg. per 100 Cm. of sonic types of cheeses fmrii may countrirs..L-” Cheese
In connection with a case of histamine intoxication after cheese,‘“, I4 we had the opportunity to study, in cooperation with the NIZO, the relationship between the histamine content of cheese and the results of scratch tests. From the appearance of this cheese, it was suspected that it had been contaminated with salt-resistant lactobacilli. Prolonged bacterial activity would lead to additional amino acid breakdown. Therefore, 2 series of cheeses were prepared, one with contarninatcd rennet and one with rennet which hat1 been purified by filtration or ccntrifugation (Table II ) There was a strong correlation between the prcscncc in chccsc ot’ amines, and histamine in particular, and a positive scratch test. The histamine concentration in the cheeses varied from 50 to 100 mg. per 100 (:rn. dry matt,er. One of t,he highest concentrations was found in the cheese which had caused the clinical
Table
I. Comprison
of
2 series
of
chreses
with
made
cw~ltanainnttd
ren?lds,
017~
series ljurificd Ilcsult Cheese
Observer
++/+++ ++ ++/+++ ++/+++ ++/+++ ++/+++
1. A” Bt 2. A B 3. A B 4. A B 5. A B *A tB
= =
contaminated. purified.
1
of swatch
tests Observw
+++ + ++ + +++ +++ +++ ++
8
Presence
of
histamine
+++++ +++++ +++++ +++ +++++ +++++ ++++
Volume Number
42 3
Histamiwe
cnusing fnlse pcwitice scratch tests
167
reaction (85 mg. per 100 cm.). Individuals varied in their reaction to histamine, but the reactions to cheese and to histamine were paralled in each subject. It is, therefore, likely that histamine present in the cheese was the cause of the local urticarial reactions. There was a cleat relationship between the presence of amincs and the bacteriological condition of the cheese. It appeared that histamine was present, especially in those cheeses prepared with rennet contaminated with salt-resistant lactobacilli. These lactobacilli withstood the increasing salt concentration rcsulting from dehydration during aging. Prolonged bacterial activity led to additional amino acid breakdown. It seerns justified to conclude that these lactobacilli were responsible for the excessive formation of histamine. COMMENTS
In some foods, histamine rnay be folnrcd by bacterial decarboxylases from histidine. Scratch tests with these pure, nonextracted foods may be influenced ha their histamine content and may lead to an unjustified diagnosis of food allergy. Although large quantities of histamine in food will be detected by scratch tests, such tests arc not scnsit,ivc enough to detect lower concentrations. Other causes of u-healing, such as the presence of histamine releasers, rnust be considered as well. Our results confirm the necessit,y of using dialyzed estract,s in skin testing, especially with foods possibly containing histamine. The likelihood that further histamine is forrned by the effect of bacterial decarboxylases on histidine after dialysis scerns slight, but warrants further study. The use of rennet c.ontaminated by salt-resistant lactobacilli in Dutch cheese is now prevented by bacttriological control by t,he I)utch Institute for* I)airy R.escarch. The authors wish to express their gratitude to the following workers for Dairy Research, Ede, The Netherlands: P. J. de Koning, Ph.D., for nations, Th. E. Galesloot, Ph.D., for the bacteriological cheese analysis, Ph.D., for literature data.
of the Dutch Institute the histamine. drtermiand C. W. Raadsveld,
REFERENCES
1. Appel, W., and Werlc1, E.: Sxchweiss van Histamin, S-dimethyl histamin, N-acstylhistamin und Actylcholin in Spin&a oleracina, Arzneimittel Forsch. 9: 22, 1959. 2. Holtz, P., and Jar&h, H.: Uher das Vorkommen van Histamin in Pflanzen, Arch. Esper. Path. Pharmacol. 187: 336, 1937. 3. Anrep, G. V., Ayadi, M. S., Barsoum, G. S., Smith, J. R., and Talaat, M. M.: The Excretion of Histamine in Urine, J. Phvsiol. 103: 155, 1944. 4. Block, R.. J.: Some A’mino &ids, Peptides and Aminrs in Milk, Concrntratetl Milks and Cheese, J. Dairy SC. 34: 1, 1951. 5. Swiatek, A., and Kisza, X.: Beitrag zur Kenntnis drr freien Diazopositivrn Aminen und Aminosauwn in Eimgen Kasesortcn, London, Proo. XV Internat. Dairy Cong. 3: 1459, 1959. G. Uusp%i, V. J., and To&i, P.: The Histamine Content of Finnish Cheese, Ann. med. exper. vt biol. fenniae 29: 58, 1951. i. lid, \v., and Kritter, H.: Zur (%cmic und Pharmakologir wrgownrr Nahrungsmittrl, I, Arch. Exper. Pharmacol. 175: 736, 1934. 8. Schullw, P. L., Ockhuizen, Th., Werringloer, J., and Marquardt, P.: Aflatoxin I& untl Histamin in Wein, Arzneimittel Forseh. 17: 888, 19867. 9. Koning, P. J. de: A New Method for thr Determination of Histamine in Cheese. To be puhlishrd. 10. Rrtkmznw, L. T., and Wilson, T. H.: A Procedure for the Determination of Histamine, Biochem. et hiophys. acta 50: 364, 1961.
Sqhnbrr, 11. Blackwell, B., ZQarley, A., and Oxydnse Inhibition, Lancct 1: 12. Geiger, IS. : Kolc of Histamine 13. Doeglas, II. J. G., Huisman, Lawet 2: 1361, 1967. 1-l. Doeglas, H. M. G., Huisman, tloor het Etcw van Kaaa, Nederl.
of Yeast Mabbitt, 1,. A. : Effects 940, 1965. in Poisoning With Spoilt,d Fish, J., and Nater, J. I’.: Histamine
Extract
After
Seic~nre 121: Intoxication
J., and Nater, .T. P.: Een Gcval v:ln tijdschr. genccwk. 111: 1526, 1967.
Histamine
J. Allergv 196.8
Monoamine865, 1955. After (%WSV, lntoxicatie
F
ood hgpersensitivities have been of recurrent interest in the allergologieal literature. In the past, highly concentrated extracts or pure foodstuffs, which frequently caused irritant-type false positive reactions, were used for scratch tests. More recently, extracts have been used which, in many cases, were dialyzed to exclude small molecule irritant materials. The causes of these irritant false positive reactions usually have remained obscure. WC have not found any studies of the relationship of the possible histamine content of foods and the outcome of scratch tests. The presence of histamine in foods may be caused by different circumstances, such as: (1) histamine in the natural state of some foods. It has been demonstrated in: spinach, concentration 2.25 mg. per 100 Gm.,] tomato, no quantita’tive data,2 and meat, concentration 0.2 to 6.0 mg. per 100 Gm.3 (2) Histamine is formed during the production of foods by the action of bacteria. Histidine in these foods undergoes decarboxylation by the microorganisms. In this case, the ba.cteria take an active part in the production proces’s of these foods: cheese, concentration 0 to 100 mg. per 100 Gm.,4-6 sauerkraut, no qua,ntitative data,’ and wine, concentration 0 to 1 mg. per 100 ml8 No doubt, the influence of time is important, and histamine will occur more readily in foods which have to go through a period of ripening or aging. (3) Histamine may be formed in the food during spoilage. Bacteria with which the food has been contaminated may form From the Department Received for publication
164
of Dermatology, Sint April 17, 1968.
Franciscus
Gnsthuis,
Rotterdam.
histamine from histidine. influence and temperature MATERIAL
AND
In this case again, duration of exposure to bacterial may play a role, for example fish and meat.
METHODS
Scratch tests were performed on 20 healthy subjects. Only pure, whole foodstuffs were used. n-0 attempt was matle at extraction or dilution. Scratches were made with a vaccinostylc, taking cart to avoid bleeding. Solid foods were gently rubbed into the scratch. Hard materials, swh as old cheese, were heated slightly, until t,hey became soft and could be smeared into the scrat,ch. In each set of tests an “empty” scratch, to exclude dermographism, and a scratch with a solut,ion of 0.1 per cent histamine HCl was included as a, cont,rol test. The study was performed by 2 independent im-cstigators in different cities. In the same 20 subjects, scratch tests were done with a number of cheeses of different origin and age. This study was done blind; it was not known beforehand which cheeses might contain histamine. The histamine content of the cheeses was determined by the I)utch Institute for Dairy &search (NIX)), with a procedure developed by I’. J. dc Koning, Ph.D. ‘2 lo It consisted of high voltage elcctrophoresis of a. watery cheese extract on thin layers of silica gel, followed by fluorimetric analysis wit,11 o-phthalaldehytle. RESULTS Various foods
Negative scratch tests were seen in all persons rubbed with pure tomato, spinach, raw meat (beef), fish (sardines, haddock), and wine (Chianti, Gcwurtz-
Fig. 1 Scratch (1) blank sauerkraut (pure), dry matter.
control, (2) histamine HCI 0.1 per and (5) cheese (pure), containing
cent in mater, (3) 85 mg. histamine
tomato HCl
(pure), per 100
166
Doe&s
J. Allergy September, 1968
and Nater
traminer, Riesling-Sylvaner) . Positive urticarial reactions of diffcrcnt strengths were seen with the following foods (Fig. 1) : 1. Marmite, Yeastrel, and the salt-free variant,, Marrnex, are yeast extracts prepared by autolysis of yeast cells in a. salt solution. Thy contain up to 200 rng. of histamine ac,id phosphate per 100 (jm.ll 2. Histamine in sa.uerkraut, has been detcrmincd only qualitatively so far,’ in some, but not all, brands of sauerkraut. 3. Sardines, directly from a, freshly opened can, gave negat,ive reactions. However, after they had been kept, in an open plastic container at room tcmperature (18’ C.) for some weeks, the reactions became strongly positive. Ceigerl’ has established that the concentra.tion of histarninc, which is present only in traces in fresh sardines, incrcascs rapidly during spoilage up to values of 120 mg. per 100 (:m. of fresh meat within 24 hours. 4. The presence of histamine ha,s been demonstrated in yuantit,ies of 5 to 100 mg. per 100 Cm. of sonic types of cheeses fmrii may countrirs..L-” Cheese
In connection with a case of histamine intoxication after cheese,‘“, I4 we had the opportunity to study, in cooperation with the NIZO, the relationship between the histamine content of cheese and the results of scratch tests. From the appearance of this cheese, it was suspected that it had been contaminated with salt-resistant lactobacilli. Prolonged bacterial activity would lead to additional amino acid breakdown. Therefore, 2 series of cheeses were prepared, one with contarninatcd rennet and one with rennet which hat1 been purified by filtration or ccntrifugation (Table II ) There was a strong correlation between the prcscncc in chccsc ot’ amines, and histamine in particular, and a positive scratch test. The histamine concentration in the cheeses varied from 50 to 100 mg. per 100 (:rn. dry matt,er. One of t,he highest concentrations was found in the cheese which had caused the clinical
Table
I. Comprison
of
2 series
of
chreses
with
made
cw~ltanainnttd
ren?lds,
017~
series ljurificd Ilcsult Cheese
Observer
++/+++ ++ ++/+++ ++/+++ ++/+++ ++/+++
1. A” Bt 2. A B 3. A B 4. A B 5. A B *A tB
= =
contaminated. purified.
1
of swatch
tests Observw
+++ + ++ + +++ +++ +++ ++
8
Presence
of
histamine
+++++ +++++ +++++ +++ +++++ +++++ ++++
Volume Number
42 3
Histamiwe
cnusing fnlse pcwitice scratch tests
167
reaction (85 mg. per 100 cm.). Individuals varied in their reaction to histamine, but the reactions to cheese and to histamine were paralled in each subject. It is, therefore, likely that histamine present in the cheese was the cause of the local urticarial reactions. There was a cleat relationship between the presence of amincs and the bacteriological condition of the cheese. It appeared that histamine was present, especially in those cheeses prepared with rennet contaminated with salt-resistant lactobacilli. These lactobacilli withstood the increasing salt concentration rcsulting from dehydration during aging. Prolonged bacterial activity led to additional amino acid breakdown. It seerns justified to conclude that these lactobacilli were responsible for the excessive formation of histamine. COMMENTS
In some foods, histamine rnay be folnrcd by bacterial decarboxylases from histidine. Scratch tests with these pure, nonextracted foods may be influenced ha their histamine content and may lead to an unjustified diagnosis of food allergy. Although large quantities of histamine in food will be detected by scratch tests, such tests arc not scnsit,ivc enough to detect lower concentrations. Other causes of u-healing, such as the presence of histamine releasers, rnust be considered as well. Our results confirm the necessit,y of using dialyzed estract,s in skin testing, especially with foods possibly containing histamine. The likelihood that further histamine is forrned by the effect of bacterial decarboxylases on histidine after dialysis scerns slight, but warrants further study. The use of rennet c.ontaminated by salt-resistant lactobacilli in Dutch cheese is now prevented by bacttriological control by t,he I)utch Institute for* I)airy R.escarch. The authors wish to express their gratitude to the following workers for Dairy Research, Ede, The Netherlands: P. J. de Koning, Ph.D., for nations, Th. E. Galesloot, Ph.D., for the bacteriological cheese analysis, Ph.D., for literature data.
of the Dutch Institute the histamine. drtermiand C. W. Raadsveld,
REFERENCES
1. Appel, W., and Werlc1, E.: Sxchweiss van Histamin, S-dimethyl histamin, N-acstylhistamin und Actylcholin in Spin&a oleracina, Arzneimittel Forsch. 9: 22, 1959. 2. Holtz, P., and Jar&h, H.: Uher das Vorkommen van Histamin in Pflanzen, Arch. Esper. Path. Pharmacol. 187: 336, 1937. 3. Anrep, G. V., Ayadi, M. S., Barsoum, G. S., Smith, J. R., and Talaat, M. M.: The Excretion of Histamine in Urine, J. Phvsiol. 103: 155, 1944. 4. Block, R.. J.: Some A’mino &ids, Peptides and Aminrs in Milk, Concrntratetl Milks and Cheese, J. Dairy SC. 34: 1, 1951. 5. Swiatek, A., and Kisza, X.: Beitrag zur Kenntnis drr freien Diazopositivrn Aminen und Aminosauwn in Eimgen Kasesortcn, London, Proo. XV Internat. Dairy Cong. 3: 1459, 1959. G. Uusp%i, V. J., and To&i, P.: The Histamine Content of Finnish Cheese, Ann. med. exper. vt biol. fenniae 29: 58, 1951. i. lid, \v., and Kritter, H.: Zur (%cmic und Pharmakologir wrgownrr Nahrungsmittrl, I, Arch. Exper. Pharmacol. 175: 736, 1934. 8. Schullw, P. L., Ockhuizen, Th., Werringloer, J., and Marquardt, P.: Aflatoxin I& untl Histamin in Wein, Arzneimittel Forseh. 17: 888, 19867. 9. Koning, P. J. de: A New Method for thr Determination of Histamine in Cheese. To be puhlishrd. 10. Rrtkmznw, L. T., and Wilson, T. H.: A Procedure for the Determination of Histamine, Biochem. et hiophys. acta 50: 364, 1961.
Sqhnbrr, 11. Blackwell, B., ZQarley, A., and Oxydnse Inhibition, Lancct 1: 12. Geiger, IS. : Kolc of Histamine 13. Doeglas, II. J. G., Huisman, Lawet 2: 1361, 1967. 1-l. Doeglas, H. M. G., Huisman, tloor het Etcw van Kaaa, Nederl.
of Yeast Mabbitt, 1,. A. : Effects 940, 1965. in Poisoning With Spoilt,d Fish, J., and Nater, J. I’.: Histamine
Extract
After
Seic~nre 121: Intoxication
J., and Nater, .T. P.: Een Gcval v:ln tijdschr. genccwk. 111: 1526, 1967.
Histamine
J. Allergv 196.8
Monoamine865, 1955. After (%WSV, lntoxicatie