Citrus flavonoids against Fusarium verticillioides in post-harvest maize: Minimization of fumonisins and alteration of fungal ultrastructure.

AIMS: To minimize fumonisins (FBs) accumulation by Fusarium verticillioides in post-harvest maize, using flavonoids obtained from citrus residues: naringin (NAR), neohesperidin (NEO), quercetin (QUER), and its mixtures. METHODS AND RESULTS: Response surface methodology with Box-Behnken design was applied in maize at 0.98 and 0.95 aw . The optimal mixture found, composed of 0.40 mmol kg-1  NAR, 0.16 mmol kg-1  NEO and 0.37 mmol kg-1 QUER, reduced the accumulation of FBs B1, B2, and B3 by 88 ± 6%, 90 ± 6% and 85 ± 5%, respectively, when applied to maize at 0.98 aw . The mentioned mixture led to a 54 ± 9% reduction of fumonisin B1 accumulation in maize adjusted to 0.95 aw . These flavonoids applied individually and as a mixture, affected the structure of both the cell wall and the cytoplasm of F. verticillioides. The cell wall lost rigidity and the cells appeared highly deformed, with ruptured plasmalemma and disrupted endomembranes. CONCLUSIONS: It was possible to diminish the accumulation of FBs in maize by a highly toxigenic Fusarium strain, producing severe damage to its ultrastructure. SIGNIFICANCE AND IMPACT OF STUDY: The results indicate the possible use of flavonoids from citrus industry residues as natural and environmentally friendly antifungal agents to restrain the accumulation of FBs in stored maize.

Evaluation of citrus flavonoids against Aspergillus parasiticus in maize: Aflatoxins reduction and ultrastructure alterations.

In this study, the effects of citrus flavonoids naringin (NAR), neohesperidin (NEO) and quercetin (QUER) on aflatoxins accumulation by a selected Aspergillus parasiticus strain in maize at 0.95 aw were studied by response surface methodology using a Box-Behnken design. Multiple response optimization was applied to simultaneously minimize the contamination with aflatoxins (AFs) B1, G1, B2 and G2. The application of the optimal mixture in maize at 0.95 aw (0.39 mM NAR, 0.24 mM NEO and 0.40 mM QUER) reduced from 85% to 100% AFs accumulation. The same mixture at 0.98 aw, led to a reduction in AFs accumulation that ranged from 93% to 98%. Ultrastructure alterations of cellular membranes and walls in A. parasiticus, evidenced by transmission electron microscopy images, were severe and depended on the type of flavonoid and their combination. Flavonoid mixtures may provide an environmentally friendly alternative for decreasing AFs accumulation in stored maize, replacing synthetic compounds.

Aflatoxins distribution in fractions derived from tofu production.

Tofu or bean curd is obtained from soybean seeds being a widespread food product in Asia. The commodity used for its production can be contaminated with aflatoxins, which are secondary metabolites synthetised by species of Aspergillus flavus and A. parasiticus. Intake of contaminated food products causes toxic effects on consumers. The aim of this work was to study aflatoxin distribution in fractions obtained from pilot-scale tofu production with contaminated soybeans. The presence of the aflatoxins B1, B2, G1 and G2 (AFs) in soaking water, okara, whey and tofu was analysed. Aflatoxin analysis was performed by HPLC with fluorescence detection. The distribution of aflatoxins in all the analysed fractions was not a normal distribution. The liquid fractions (soaking water and whey) had less contamination than solid fractions (tofu and okara). The percentage AFB1 remaining in nutritionally important fractions, okara and tofu, was between 6.2% and 67.7% (median = 18.1%) and 0.5% and 13.2% (median = 3.5%), respectively. AFB2, AFG1 and AFG2 had a similar distribution. These results showed that throughout tofu production, AFs can be present in the products intended for human consumption.

Factores extrínsecos e intrínsecos asociados a poblaciones fúngicas micotoxigénicas de granos de maíz (Zea mays L) almacenados en silos bolsa en Argentina / Extrinsic and intrinsic factors associated with mycotoxigenic fungi populations of maize grains (Zea mays L.) stored in silobags in Argentina

Con el objeto de caracterizar las poblaciones fúngicas, en particular las especies potencialmente micotoxigénicas, que pueden contaminar los granos de maíz almacenados en silos bolsa con un contenido de humedad superior al recomendado como seguro, se evaluaron 270 muestras extraídas al inicio, a los 90 días y al final de un período de almacenamiento de 5 meses. En dichas muestras se cuantificó e identificó la biota fúngica y se determinó la contaminación con fumonisinas y aflatoxinas. Asimismo, se evaluó el efecto de factores extrínsecos (ambiente), intrínsecos (granos) y tecnológicos (ubicación de los granos en el perfil del silo bolsa) sobre las poblaciones totales y micotoxigénicas. El pH de los granos y el nivel de O2 se redujeron significativamente a los 5 meses, mientras que la concentración de CO2 se incrementó en igual período. Los recuentos totales de la micobiota fueron significativamente mayores en los granos ubicados en el estrato superior del silo bolsa. Se identificaron especies micotoxigénicas de Fusarium, Aspergillus, Penicillium y Eurotium. La frecuencia de aislamiento de Fusarium verticillioides se redujo al final del almacenamiento y Aspergillus flavus solo se aisló en el inicio del almacenamiento. Los recuentos de Penicillium spp. y Eurotium spp. se incrementaron al final del almacenamiento. El 100 % de las muestras presentaron contaminación con fumonisinas, con niveles máximos de 5,707 mg/kg, mientras que las aflatoxinas contaminaron el 40 % de las muestras con niveles máximos de 0,0008 mg/kg. Las condiciones ambientales y de sustrato generadas durante el almacenamiento produjeron cambios en la composición de las poblaciones fúngicas y limitaron el desarrollo de hongos micotoxigénicos y la producción de micotoxinas.

In order to determine the behavior of mycotoxin-producing fungal populations linked with silobags stored corn grains with a moisture content greater at the recommended as safe, 270 samples taken in three times (beginning, 90 days, final) over a five month period of storage were evaluated. The fungal biota was quantified and identified and the contamination with fumonisin and aflatoxin was determined. Extrinsic factors (environment), intrinsic factors (grains) and technological factors (location of the grains in the profile of silobag) were taken into account to evaluate the presence and quantity of total and mycotoxigenic fungal populations. The pH of grains and O2 levels were significantly reduced after five months, while CO2 concentration increased in the same period. The total counts of mycobiota were significantly higher in grains located in the top layer of silobag. Mycotoxigenic species of Fusarium, Aspergillus, Penicillium and Eurotium were identified. The frequency of isolation of Fusarium verticillioides decreased at the end of storage and Aspergillus flavus was isolated only at the beginning of storage. The counts of the Penicillium spp. and Eurotium spp. were increased at the end of storage. Fumonisin contamination was found in all the samples (100 %) with maximum levels of 5.707 mg/kg whereas aflatoxin contaminated only 40 % with maximum levels of 0.0008 mg/kg. The environmental and substrate conditions generated during the storage limited the development of mycotoxigenic fungi and mycotoxin production.

[Extrinsic and intrinsic factors associated with mycotoxigenic fungi populations of maize grains (Zea mays L.) stored in silobags in Argentina]. / Factores extrínsecos e intrínsecos asociados a poblaciones fúngicas micotoxigénicas de granos de maíz (Zea mays L.) almacenados en silos bolsa en Argentina.

In order to determine the behavior of mycotoxin-producing fungal populations linked with silobags stored corn grains with a moisture content greater at the recommended as safe, 270 samples taken in three times (beginning, 90 days, final) over a five month period of storage were evaluated. The fungal biota was quantified and identified and the contamination with fumonisin and aflatoxin was determined. Extrinsic factors (environment), intrinsic factors (grains) and technological factors (location of the grains in the profile of silobag) were taken into account to evaluate the presence and quantity of total and mycotoxigenic fungal populations. The pH of grains and O2 levels were significantly reduced after five months, while CO2 concentration increased in the same period. The total counts of mycobiota were significantly higher in grains located in the top layer of silobag. Mycotoxigenic species of Fusarium, Aspergillus, Penicillium and Eurotium were identified. The frequency of isolation of Fusarium verticillioides decreased at the end of storage and Aspergillus flavus was isolated only at the beginning of storage. The counts of the Penicillium spp. and Eurotium spp. were increased at the end of storage. Fumonisin contamination was found in all the samples (100%) with maximum levels of 5.707mg/kg whereas aflatoxin contaminated only 40% with maximum levels of 0.0008mg/kg. The environmental and substrate conditions generated during the storage limited the development of mycotoxigenic fungi and mycotoxin production.

Mycotoxigenic potential of fungi isolated from freshly harvested Argentinean blueberries.

Alternaria alternata, A. tenuissima, Fusarium graminearum, F. semitectum, F. verticillioides, Aspergillus flavus, and Aspergillus section Nigri strains obtained from blueberries during the 2009 and 2010 harvest season from Entre Ríos, Argentina were analyzed to determine their mycotoxigenic potential. Taxonomy status at the specific level was determined both on morphological and molecular grounds. Alternariol (AOH), alternariol monomethyl ether (AME), aflatoxins (AFs), zearalenone (ZEA), fumonisins (FBs), and ochratoxin A (OTA) were analyzed by HPLC and the trichotecenes deoxynivalenol (DON), nivalenol (NIV), HT-2 toxin (HT-2), T-2 toxin (T-2), fusarenone X (FUS-X), 3-acetyl-deoxynivalenol (3-AcDON), and 15-acetyl-deoxynivalenol (15-AcDON) by GC. Twenty-five out of forty two strains were able to produce some of the mycotoxins analyzed. Fifteen strains of Aspergillus section Nigri were capable of producing Fumonisin B1 (FB1); two of them also produced Fumonisin B2 (FB2) and one Fumonisin B3 (FB3). One of the F. graminearum isolated produced ZEA, HT-2, and T-2 and the other one was capable of producing ZEA and DON. Two A. alternata isolates produced AOH and AME. Four A. tenuissima were capable of producing AOH and three of them produced AME as well. One Aspergillu flavus strain produced aflatoxin B1 (AFB1), aflatoxin B2 (AFB2), and aflatoxin G1 (AFG1). To our knowledge, this is the first report showing mycotoxigenic capacity of fungal species isolated from blueberries that include other fungi than Alternaria spp.

Mycoflora and mycotoxin contamination of Roundup Ready soybean harvested in the Pampean Region, Argentina.

A total of 89 freshly harvested soybean seed samples (Roundup Ready [transgenic] soybean cultivars) from the 2010/2011 crop season were collected from five locations in the Northern Pampean Region II, Argentina. These samples were analyzed for internal mycoflora, toxin production of isolated fungi, and for a range of mycotoxins. Mycotoxin analysis of aflatoxins (AFs), zearalenone (ZEA), fumonisins (FBs) and ochratoxin A (OTA) was done by HPLC-FLD (high performance liquid chromatography with postcolumn fluorescence derivatization), alternariol and alternariol monomethyl ether with HPLC-UV (HPLC with UV detection), trichothecenes (deoxynivalenol, nivalenol, T-2 toxin, HT-2 toxin, fusarenon X, 3-acetyldeoxynivalenol and 15-acetyldeoxynivalenol were analyzed by GC-ECD (gas chromatography with electron capture detector). Fungal colonization was more frequently found for samples from América, Saladillo and Trenque Lauquen than for samples from General Villegas and Trenel; a total of 1,401 fungal isolates were obtained from the soybean seeds. The most commonly identified fungal genera were Alternaria, Sclerotinia, Chaetomium, Cladosporium, Aspergillus, Penicillium, Phomopsis and Fusarium. Alternaria alternata, A.tenuissima, Aspergillus flavus, Penicillium citrinum, Fusarium verticillioides and F.semitectum were the predominant toxigenic fungal species. Mycotoxin production was confirmed for several isolates of toxigenic species, including Aspergillus flavus, A. parasiticus, Alternaria alternata, A.tenuissima, Fusarium graminearum, F semitectum and F. verticillioides. In particular, the percentage of mycotoxigenic Alternaria alternata (100%), A.tenuissima (95%) and aflatoxigenic strains of A. flavus (57%) were remarkably high. Although none of the mycotoxins, AFs, ZEA, FBs, trichothecenes and OTA, were directly detected in samples of soybean seeds, the frequent presence of toxigenic fungal species indicates the risk of multiple mycotoxin contamination.

Equine leukoencephalomalacia (ELEM) due to fumonisins B1 and B2 in Argentina / Leucoencefalomalacia equina devida a fumonisinas B1 e B2 na Argentina

In August 2007 an outbreak of neurological disease and sudden death in Arabian horses occurred in a farm located in Coronel Rosales County, Buenos Aires Province, Argentina. The animals were on a pasture of native grasses and supplemented ad libitum with corn kernels and wheat bran. Three horses were observed having acute neurologic signs including blindness, four leg ataxia, hyperexcitability, aimless walking and circling, followed by death in two of them. Four other horses were found dead overnight without a history of neurologic signs. The morbidity, mortality and lethality rates were 11.6 percent, 10 percent and 85.7 percent, respectively. Grossly, the brain showed focal areas of hemorrhage, brown-yellow discoloration and softening of the sub-cortical white matter. The microscopic brain lesions consisted of extensive areas of malacia within the white matter of the cerebral hemispheres, brainstem and cerebellum, characterized by rarefaction of the white matter with cavitations filled with proteinaceous edema, multifocal hemorrhages and mild infiltration by neutrophils, and rare eosinophils. Swollen glial cells with abundant eosinophilic cytoplasm, distinct cell borders, intracytoplasmic deeply eosinophilic globules and eccentric, hyperchromatic, occasionally pyknotic nucleus were present throughout the areas of rarefaction hemorrhage, edema and necrosis. The feed supplements contained 12,490µg/kg of fumonisin B1 and 5,251µg/ kg of fumonisin B2. This is the first reported outbreak of ELEM associated with consumption of feed supplements containing high concentrations of fumonisins in Argentina.

Em agosto de 2007, ocorreu um surto de doença neurológica e morte súbita em cavalos árabes em uma propriedade localizada no município de Coronel Rosales, na província de Buenos Aires, Argentina. Os animais estavam em pasto nativo e eram suplementados ad libitum com grãos de milho e farelo de trigo. Três cavalos foram observados com sinais neurológicos agudos, incluindo cegueira, ataxia nas quatro pernas, hiperexcitabilidade, e andar sem rumo e em círculo, seguidos de morte em dois animais. Outros quatro cavalos foram encontrados mortos durante a noite sem histórico de distúrbios neurológicos. A mortalidade, morbidade e letalidade foram de 11,6 por cento, 10 por cento e 85,7 por cento, respectivamente. Macroscopicamente, o cérebro tinha áreas focais de hemorragia, coloração amarelada e amolecimento da substância branca sub-cortical. Microscopicamente, as lesões cerebrais consistiram de extensas áreas de malácia na substância branca dos hemisférios cerebrais, do tronco encefálico e do cerebelo. Estas lesões da substância branca se caracterizaram por rarefação, cavidades contendo fluido proteináceo (edema), hemorragias multifocais e moderado infiltrado por neutrófilos e raros eosinófilos. Células gliais tumefeitas com abundante citoplasma eosinifílico, limites celulares evidentes, globules citoplasmáticos eosinofílicos, e núcleo excéntrico, hipercromático e ocasionalmente picnótico foram observadas nas areas de rarefacção, edema, hemorragias e necrose. Os suplementos alimentares continham 12.490µg/kg de fumonisina B1 e 5.251µg/ kg de fumonisina B2. Este é o primeiro surto reportado na Argentina de leucoencefalomalácia equina associado ao consumo de suplementos alimentares contendo altas concentrações de fumonisinas.

Fungal and fumonisins contamination in Argentine maize (Zea mays L.) silo bags.

Fumonisins in maize (Zea mays L.) grain silo bags in the conditions of three Argentine provinces were analyzed to determine how this kind of storage affects contamination and if differential storage durations or times of the year of silo bag closing and opening are factors that could modify it. Moisture content, water activity (a(w)), molds and yeasts present, and fumonisins were analyzed in 163 maize silo bags, at the moment of closing and later at opening. Storage durations ranged from 120 to 226 days. The analysis was centered on fumonisins since most samples were only contaminated with these toxins. Fumonisins, moisture content, and a(w) increased significantly, whereas mold propagules/g and yeasts CFU/g did not present significant differences between silo opening and closing. The date on which silo bags were closed and later opened, however, did affect the level of fumonisin contamination.

Alternaria alternata prevalence in cereal grains and soybean seeds from Entre Ríos, Argentina.

A mycological survey was carried out at Entre Ríos province, Argentina, on sorghum grain, maize, rice, soybean seeds and on freshly harvested and stored wheat. The isolation frequencies and relative densities of species belonging to genera Alternaria, Aspergillus, Fusarium, Penicillium and other fungi were calculated. Alternaria alternata was the major fungal species isolated from sorghum, rice, soybean seeds and on freshly harvested wheat, and a low incidence of Fusarium species was observed on the same substrates. In maize the major fungal species isolated was Fusarium verticillioides. The high incidence levels of A. alternata observed,suggest that it may be necessary to determine, among other mycotoxins, if Alternaria toxins occur in these commodities.

Alternaria alternata prevalence in cereal grains and soybean seeds from Entre Ríos, Argentina

En la provincia de Entre Ríos, Argentina se llevo a cabo un estudio micológicoen granos de sorgo, maíz, arroz y semillas de soja y en muestras de trigorecién cosechado y con un año de almacenamiento. Se calcularon lasfrecuencias de aislamiento y las densidades relativas específicas de especiespertenecientes a los géneros Alternaria, Aspergillus, Fusarium, Penicilliumy otros. Alternaria alternata, fue la especie más aislada en sorgo, arroz,soja y en el trigo recién cosechado. En los mismos sustratos se observó unaincidencia menor de especies de Fusarium. En maíz, la especie fúngicapredominante fue Fusarium verticillioides. Los altos niveles de incidencia deA. alternata observados, indicarían la necesidad de determinar, entre otrasmicotoxinas, la presencia natural de toxinas de Alternaria en estos sustratos(AU)

A mycological survey was carried out at Entre Ríos province, Argentina,on sorghum grain, maize, rice, soybean seeds and on freshly harvested andstored wheat. The isolation frequencies and relative densities of speciesbelonging to genera Alternaria, Aspergillus, Fusarium, Penicillium and otherfungi were calculated. Alternaria alternata was the major fungal speciesisolated from sorghum, rice, soybean seeds and on freshly harvested wheat,and a low incidence of Fusarium species was observed on the samesubstrates. In maize the major fungal species isolated wasFusarium verticillioides. The high incidence levels of A. alternata observed,suggest that it may be necessary to determine, among other mycotoxins,if Alternaria toxins occur in these commodities(AU)

Distribution of deoxynivalenol in wheat, wheat flour, bran, and gluten, and variability associated with the test procedure.

Analytical data obtained on deoxynivalenol (DON) concentration in naturally contaminated wheat during processing in an industrial mill were statistically analyzed, and the distribution functions of DON concentration in lots of wheat, bran, wheat flour, and gluten were estimated. The analytical method had acceptable precision (HORRAT 0.25-0.32) for each test sample. The total variance combined sampling, sample preparation, and analytical variances were 0.188, 0.033, 0.42, and 0.0014 ppm2 for wheat, 1.93; flour, 0.99; bran, 4.68; and gluten, 0.29, respectively. The distribution function of DON contamination presented an asymmetric tail for high values of concentration in wheat grains and wheat flour; in bran it seemed to be bimodal with 2 separated peaks of different concentrations; in gluten the normal distribution function gave a reasonably good fit to empirical data. The function eta(c) = -In(-Inp), where p (c) is the cumulative distribution function was linear with c in the so-called extreme-value type I distribution and could be fitted by a cubic polynomial in c in the distributions determined for all the products. This variability and distributional information contributes to the design of better sampling plans in order to reduce the total variability and to estimate errors in the evaluation of DON concentration in lots of wheat and wheat products.

Consumo de alimentos e ingesta de algunos nutrientes en la población de la Universidad Nacional de Luján, Argentina / Food comsumption and intake of several nutrients of Luján University's population, Argentina

Se llevó a cabo en la población de la Universidad Nacional de Luján una encuesta dietética de recordatorio de 24 hs. Para evaluar: a) el consumo de alimentos y el aporte energético efectuado por el grupo de cereales, y b) la adecuación de la ingesta de proteínas, calcio, hierro, vitaminas A, B1, B2, C y niacina. La muestra (16 por ciento de la población y 4 por ciento en cada una de las estaciones del año) estuvo compuesta por: 189 varones (G1V) y 209 mujeres (G1M) de 18 a 24 años; y 189 varones (G2V) y 240 mujeres (G2M), entre 25 y 50 años. El aporte energético de los cereales (90 por ciento trigo) fue 32 por ciento en las mujeres jóvenes, entre el 40 por ciento y 48 por ciento en los otros tres grupos. El consumo promedio diario de carne vacuna osciló entre 90,5 y 128,7 g en las mujeres y fue superior a 140,0 g en los varones. El consumo de productos lácteos, así como el de frutas y verduras fue muy bajo en todo los grupos. La ingesta promedio de proteínas fue superior a 1,25 g/kg/día en el 50 por ciento de la población total. La ingesta de calcio fue inferior a la ingestas recomendadas en el 71 por ciento de G1M. 62 por ciento de G2M, 64 por ciento de G1V y 65 por ciento de G2V. Las ingestas promedio de hierro oscilaron entre 16,4 y 20,8 mg/día en las mujeres y 17,5 y 19,2 mg/día en los varones, siendo aportado por las carnes: G1M 16 por ciento, G2M 21 por ciento, G1V 34 por ciento y G2V 26 por ciento (lo que implica una biodisponibilidad elevada), y por las infusiones, fundamentalmente el mate (56 por ciento en G1M, 12 por ciento en G1V, 25 por ciento G2V y 29 por ciento G2M). La ingesta de vitamina A superó las ingestas recomendadas entre el 26 por ciento y el 41 por ciento en los diferentes grupos. La ingesta promedio de vitamina B1 representó el 89 por ciento de las ingestas recomendadas en los varones jóvenes y las superó en los demás grupos. La ingesta promedio de vitamina B2 superó las ingestas recomendadas en todos los grupos, con una gran disposición de valores. Este comportamiento se repitió en el caso de la vitamina C y la niacina. Los resultados expresados muestran un elevado consumo de proteínas, bajo consumo de láteos, lo que trae aparejado una insuficiente ingesta de calcio y vitamina A, y bajo consumo de frutas, verduras y hortalizas. Además, desde el punto de vista toxicológico, el predominio del consumo de alimentos derivados del trigo alerta...

La funcion hepatica a traves del espectro clinico de la hanseniasis / The hepatic function through the clinical phantom of the leprosy

Se estudio la funcion hepatica em 100 pacientes hansenianos agrupados seguin la classificacion especial y la presencia o no de episodios reacionales del seguiente modo: pacientes quiescentes: 20 LL, 13 BL, 7 BB, 9 BT, 12 TT. Pacientes reaccionales: 12 LL, 10 BL, 7 BB, 5 TT. Como grupo testigo se estudiaron 10 individuos sanos no contactantes com pacientes hansenianos y sin antecedentes de patologia hepatica. Los resultados obtenidos muestran una hiperproteinemia a expensas de las globulinas sin disminucion de albumina en todas las formas clinicas de la hanseniase reaccionales y quiescentes, y un aumento de la actividad enzimatica serica de GPT, GOT y fosfatasa alcalina en los pacientes reaccionales. Estos incrementos de la actividade enzimatica caracterizan un duno hepatocelular leve, en que la hilirruhinemia y la prueba de turbidez del timol no presentan alteraciones. Se discuten los mecanismos fisiopatologicos causantes de estas alteraciones, postulindos su vinculacion com fenomenos de hipersensibilidad a tipo III y IV.

Perfil de las isoenzimas de la dehidrogenasa lacta en el suero de pacientes hansenianos / Profile of isoenzymes of dehidrogenasa lactica in the serum of leprosy pacients

Se midieron los niveles sericos de las isoenzimas de la dehidrogensas lactica (LDH) en siete pacientes de hanseniasis de la Republica Argentina. El fraccionamiento se hizo por electroforesis en gel de poliacrilamida. En todos los casos la LDH-2 resulto disminuida y la LDH-4 aumentada; la fraccion LDH-5, que fue detectada en solo tres de los casos, resulto muy elevada com respecto a los valores normales.

Estudios enzimaticos en pacientes hansenianos. I. Actividade de las transminasas glutamico piruvica y glutamico oxacetica / Enzymatic studies in leprosy patients. Activity of transaminasas glutámico pirúvica and oxalacética glutámico

Se estúdio la actividad enzimática de Got y GPT em 130 pacientes hansenianos divididos em cuatro grupos: hanseniasis virchowiana quiescente, tuberculoide quiescente, virchowiana reaccional y tuberculoide reaccional. Se observaron diferencias significativas entre lãs formas quiescentes y lãs reaccionales; no se observo diferencia entre lãs formas quiescentes entre si, em tanto que la actividad enzimática em los pacientes virchowianos reaccionales estava elevada com respecto a los tuberculoides reaccionales em forma poço significativa. Se discute que la causa de estas alteraciones se deba a fenômenos de hipersensibilidad mediados por inmunocomplejos o por células segun los casos