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Rev Biomed 1999; 10:29-31.

Presence of Listeria monocytogenes in fresh salad vegetables.

Brief Communication

Rafael Monge1, Mara L. Arias-Echandi 2.

Instituto Costarricense de Investigacin y Enseanza en Nutricin y Salud, 2Facultad de Microbiologa, Universidad de Costa Rica. San Jos, Costa Rica.

SUMMARY. Introduction. An increase in the consumption of fresh vegetables for a healthy diet has ocurred during the last few years, but it mustnt be forgotten that they can carry pathogenic microorganisms such as Listeria monocytogenes. Material and methods. The presence of Listeria monocytogenes was evaluated in 50 salad samples according to the methodology described by Vanderzant & Splittstoesser. Results. Listeria spp. was isolated from 32% (16/50) of the samples analyzed, from these, 8% (10/16) were L. monocytogenes, 8% (4/16) a L. welshimeri and 4% (2/16) L. murrai. Discussion The presence of Listeria monocytogenes in fresh salad vegetables represents a potential risk to public health, since its infective dose has not been established yet and its ability to survive and grow in raw vegetables has been demonstrated. Dietary changes should be promoted in groups where this bacteria represents a risk. (Rev Biomed 1999; 10:29-31)

Key words: Listeria spp., Listeria monocytogenes, salad, vegetables.

RESUMEN. Presencia de Listeria monocytogenes en vegetales frescos. Introduccin. En los ltimos aos, se ha dado un incremento en el hecho de consumir vegetales frescos como parte de una dieta sana, pero no se puede descartar el que estos pueden actuar como vehculos de microorganismos patgenos, como Listeria monocytogenes. Material y mtodos. Se evalu la presencia de Listeria monocytogenes en 50 muestras de ensalada, segn la metodologa descrita por Vanderzant y Splittstoesser. Resultados. Se aisl Listeria spp. a partir de 32% (16/50) de las muestras analizadas, de stas, 8% (10/16) correspondi a L. monocytogenes, 8% (4/16) a L. welshimeri y 4% (2/16) a L. murrai.

Corresponding address: Mara L. Arias-Echandi. Facultad de Microbiologa, Universidad de Costa Rica. Ciudad Universitaria "Rodrigo Facio", San Jos, Costa Rica. Email: [email protected] Received April 2, 1998; Accepted June 30, 1998. This paper is also available at http://www.uady.mx/~biomedic/rb991015.html

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R Monge, ML Arias-Echandi. Discusin. La presencia de L. monocytogenes en ensalada de vegetales frescos representa un riesgo potencial para la salud pblica, dado que su dosis infectante no ha sido claramente definida y est demostrada su capacidad de sobrevivir en vegetales crudos. Se debe de promover cambios de dieta en grupos donde esta bacteria representa un riesgo. (Rev Biomed 1999; 10:29-31) Palabras clave: Listeria spp., Listeria monocytogenes, ensalada, vegetales. presence of L. monocytogenes in fresh salad vegetables sold in the metropolitan area of San Jos, Costa Rica.

INTRODUCTION. During recent years there has been an increasing emphasis on the importance of the consumption of fresh vegetables for a healthy diet. Fresh vegetables are an important source of fiber, carothenes, vitamin C and other nutrients, but they can also carry pathogenic microorganisms. Seven species of Listeria spp. have been identified, but L. monocytogenes is the main pathogen for both humans and animals. The symptoms of the disease range from a mild flu-like illness to a severe septicaemia, particularly in children the elderly and in the immunosupressed; infection during pregnancy can result in abortion, stillbirth or birth of a severely affected baby (1). L. monocytogenes is a widely distributed bacteria in nature (2), so plants play an important role in its dissemination (3). This bacteria can reach humans either indirectly like ingesting milk from animals that had consumed contaminated food, directly in the form of contaminated foods, especially vegetables and dairy products (3). Several listeriosis outbreaks have been associated to the consumption of raw vegetables, especially raw celery, tomatoes, lettuce, cabbage and coleslaw (4). Occurrence of L. monocytogenes in fresh salad vegetables has been reported in industrialized countries, however little is known about the incidence of this bacteria in this kind of food in other nations. The aim of this study was to evaluate the Revista Biomdica

MATERIAL AND METHODS. A total of 50 samples of fresh salad vegetables, randomly acquired, was analyzed during August and September, 1997. 50% of the samples represented salads served as part of a popular dish known as "casado", the other 50% were prepackaged salads sold in the supermarkets. Samples were tested for the presence of Listeria spp. according to the methodology recommended by Vanderzant & Splittstoesser (5). Briefly, 25 g of each sample were transferred to 225 ml UVM broth (University of Vermont modified Listeria enrichment broth) and incubated for 22+/- 2 h at 30C. After this, 1 ml was transferred to 10 mL Fraser broth and incubated at 37 C for 24 +/- 2 h. After the enrichment, a loop was streaked on the surface of Oxford agar, incubated at 37 C for 48 h and evaluated for the presence of typical Listeria colonies. These were confirmed by Henry illumination, morphology and Gram staining, motility, hemolysis properties, CAMP (Chriestie, Atkins and Munch-Peterson) behavior with Staphylococcus aureus , xylose and rhamnose utilization.

RESULTS. Listeria was found in 32% (16/50) of the samples analyzed. From these, 20% (10/16) were L. monocytogenes , 8% (4/16) L. welshimeri and 4% (2/16) L. murrai. 100% of the samples positive for L. monocytogenes had cabbage as the base vegetable.

DISCUSSION. The incidence of L. monocytogenes in fresh vegetable salads is quite variable, it can be as low as 1.8% in North London (6), 4.5% in Switzerland

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Listeria monocytogenes in fresh salad vegetables. (7), or as high as 60% in the US (3). In spite of this variable pattern, fresh salad vegetables are becoming a matter of concern as a potential source of L. monocytogenes in human infections (8). Even though in this study the presence of L. monocytogenes was not quantified, it's mere presence in fresh salad vegetables constitutes a potential health risk, especially for immunocompromised individuals, including pregnant women and the elderly, since its infective dose has not yet been established and its ability to survive and grow in raw vegetables destined for human consumption has been widely and adequately demonstrated. L. monocytogenes is known to survive in plant vegetation for 10 to 12 years and it can reach populations of 105 to 10 7 CFU/g when stored at 4-15 C respectively (3). Bracket (9) also reports that this organism has remained viable on Brussel sprouts dipped for 10 s in water containing 200 ug/ml chlorine. Our results suggest the need for promoting changes in dietary habits for pregnant women and other immunosupressed groups, especially when it is considered that the role of L. monocytogenes in raw vegetables is still not clearly understood. We recommend substituting the ingestion of raw vegetables for boiled ones, in which the thermic treatment destroys pathogenic microorganisms and also increases the presence of some nutritive elements as dietary fiber.
ACKNOWLEDGEMENTS. We wish to thank Laura Villalobos for her cooperation. 4.- Farber J, Peterkin P. Listeria monocytogenes, a food borne pathogen. Microbiol Rev 1991; 55: 476-511. 5.- Vanderzant C, Splittstoesser D. Compendium of methods for the microbiological examination of foods. Washington: APHA; 1992. p. 317. 6.- Velani S, Roberts D. Listeria monocytogenes and other Listeria spp. in prepacked mixed salads and individual salad ingredients. PHLS Microbiol Digest 1991; 8:21-2. 7.- Sizmur K, Walker C. Listeria in prepacked salads. Lancet 1988; 1:1167. 8.- Steinbruegge E, Burt R, Liewen M. Fate of Listeria monocytogenes on ready to serve lettuce, J Food Prot 1988; 51:596-9. 9.- Brackett R. Antimicrobial effect of chlorine on Listeria monocytogenes. J Food Prot 1987; 50:999-1003.

REFERENCES. 1.- Todd E. Food-borne illness. Lancet 1990; 336:78893. 2.- Weis J, Seeliger KPR. Incidence of Listeria monocytogenes in nature. J Appl Bacteriol 1975; 50:1-9. 3.- Beuchat L, Berrang H, Brackett R. Presence and public health implications of Listeria monocytogenes on vegetables In: Miller, AJ, Smith L, Somkusti G, (editores). Foodborne listeriosis. New York: Elsevier Science; 1990. p. 175-81.

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