First draft prepared by
    Dr Margaret F.A. Wouters and Dr G.J.A. Speijers
    Laboratory for Toxicology, National Institute of Public Health and
    Environmental Protection, Bilthoven, Netherlands

    Biological data
         Biochemical aspects
         Absorption, distribution, and excretion
         Effects on enzymes and other biochemical parameters
    Toxicological studies
         Acute toxicity studies
         Short-term toxicity studies
         Long-term toxicity/carcinogenicity studies
         Reproductive toxicity studies
         Special studies on antibiotic activity
         Special studies on antitumour activity
         Special studies on cytotoxicity
         Special studies on genotoxicity
         Special studies on immunotoxicity
         Special studies on neurotoxicity
         Special studies on teratogenicity and embryotoxicity
         Observations in humans


         Patulin is a mycotoxin produced by certain species of the genera
     Aspergillus and  Penicillium, including  A. clavatus, P. expansum, P.
     patulum, P. aspergillus and  P. byssochlamys. P. expansum is a common
    spoilage microorganism in apples, and the major potential dietary
    sources of patulin are apples and apple juice made from affected fruit.

         Patulin was previously evaluated by the Committee at its
    thirty-fifth meeting (Annex 1, reference 88), when a PTWI of 7 g/kg
    bw was established based on a no-effect level of 0.1 mg/kg bw/day in a
    combined reproductive toxicity/long-term toxicity/carcinogenicity
    study in rats. Additional information has become available since the
    last evaluation.

         Patulin was reviewed by IARC (IARC, 1976; 1985). It was concluded
    at the second of these reviews that there was inadequate evidence for
    carcinogenicity of patulin in experimental animals. No evaluation
    could be made of carcinogenicity of patulin in humans.

         The following toxicological monograph summarizes both the
    information given in the previous toxicological monograph and
    information received since the previous review.


    2.1  Biochemical aspects

    2.1.1  Absorption, distribution, and excretion

         A single oral dose of 3 mg/kg bw of 14C-patulin in citrate
    buffer was given to 17 male and 12 female Sprague-Dawley rats exposed
    for 41-66 weeks after birth to levels of 0 or 1.5 mg/kg bw of patulin
    in 1 mol/litre citrate buffer. All animals were fasted for 24 h before
    the administration of the labelled patulin. Animals were placed in
    metabolic cages and faeces, urine and CO2 were collected. One or 2
    animals/sex/group (untreated or pretreated with patulin) were
    sacrificed at 4, 24, 48, 72 h or 7 days after blood was collected for
    patulin determination. Concentrations of patulin in erythrocytes were
    calculated from the difference between radioactivity of whole blood
    and serum. Within 7 days, 49% and 36% of the administered radio-
    activity was recovered in faeces and in urine, respectively. Most of
    the excretion of label occurred within the first 24 h. Approximately
    1-2% of the label was recovered as 14CO2. At the end of 7 days,
    2-3% of the radioactivity was recovered in soft tissues and blood. The
    major retention sites of patulin were erythrocytes and blood-rich
    organs (spleen, kidney, lung and liver) (Dailey  et al., 1977a).

    2.1.2  Effects on enzymes and other biochemical parameters

     In vivo studies

         Absorption of radiolabelled glycine, alanine and lysine was
    reduced in perfused intestines of rats that had received 100 mg of
    patulin intraperitoneally on alternate days for 1 month (equal to
    1.6 mg/kg bw/day). The authors attributed this effect to reduced total
    ATPase, NaK ATPase, and alkaline phosphatase activities which were
    studied in a satellite group of rats (Devaraj  et al., 1982a).

         A group of albino rats received 0.1 mg of patulin in 0.2 ml of
    propylene glycol, injected i.p. on alternate days. A control group was
    treated similarly with propylene glycol alone. The animals were
    sacrificed after 15 doses. Liver, kidney and intestine were used for
    the assay of various enzymes such as glycogen phosphorylase,
    hexokinase, glucose-6-phosphatase, fructose-1,6-diphosphorylase,
    hexokinase, glucose-6-phosphatase, fructose-l,6-diphosphatase and

         The concentration of aldolase in the liver, kidney and intestinal
    tissue was reduced during patulin toxicosis. In a follow up
    experiment, groups of rats were treated similarly and the rats were
    sacrificed after 20 doses. Aldolase was isolated and purified and
    studies on its kinetic properties were made. These studies did not
    show any significant variations in the properties of liver aldolase of
    normal and patulin-treated rats. The authors concluded that the
    results suggested that patulin toxicosis inhibited the biosynthesis of
    liver aldolase (Sakthisekaran & Shanmugasundaram, 1990).

         Forty-eight hours after i.p. injection of 5.0 or 7.5 mg/kg bw of
    patulin in male ICR mice, NaKATPase and MgATPase of liver, kidney and
    brain preparations were significantly inhibited. Injection of
    2.5 mg/kg bw had no significant effect on enzyme activity. The same
    effects were demonstrated in  in vitro studies with mitochondrial and
    microsomal fractions of liver, kidney and brain of ICR mice (Phillips
    & Hayes, 1977).

         Patulin inhibited acetylcholinesterase and NaKATPase in cerebral
    hemisphere, cerebellum and medulla oblongata in rats treated for 1
    month with i.p. injections of 1.6 mg/kg bw/day patulin. Concomitantly,
    acetylcholine levels were raised in these brain segments (Devaraj
     et al., 1982b).

         A non-competitive and irreversible inhibition of the activity of
    alcohol dehydrogenase derived from yeast was attributed to patulin's
    ability to bind to SH-groups; the K1 was found to be 5.0  10-5 M
    (Ashoor & Chu, 1973a).

         Inhibition of yeast-derived aminoacyl-tRNA synthetase by patulin
    was mainly due to modification of the enzyme's sulfhydryl groups
    (Arafat,  et al., 1985).

         Liver lactate dehydrogenase was increased in 4 pregnant
    Sprague-Dawley rats after exposure by gavage to 3 mg/kg bw/day of
    patulin in tris-acetate buffer, from days 1-19 of gestation
    (Fuks-Holmberg, 1980).

         Malate dehydrogenase in human placental microsome- and
    mitochondria-rich fractions was increased up to 15 times when
    incubated with 0.5 - 3 mg/g placenta of patulin  in vitro
    (Fuks-Holmberg, 1980).

         Placental GPT was depressed in 4 pregnant Sprague-Dawley rats
    after exposure by gavage to 3 mg/kg bw/day of patulin in tris-acetate
    buffer, from days 1-19 of gestation (Fuks-Holmberg, 1980).

         When white male albino mice were injected with 10 doses of
    0.1 mg of patulin in propylene glycol on alternate days, glycogen
    phosphorylase in the liver was activated, and blood glucose levels
    increased by 60%. These results were confirmed by  in vitro studies
    (Madiyalakan & Shanmugasundaram, 1978).

         Groups of 10 rats were fed regular diet, diet infected with
     Penicillium patulum, or received i.p. injections of purified patulin
    (1 mg/kg bw on alternate days) for 3 months. Fasting blood glucose
    levels were elevated and a glucose tolerance test revealed an elevated
    glucose curve and reduced insulin production. The authors concluded
    that patulin was diabetogenic (Devaraj  et al., 1986).

         Male F344 rats received a single i.p. injection of 0, 0.5, 5 or
    10 mg/kg bw patulin. Liver mixed function oxidase and cytochrome P-450
    activity were determined 4 days after treatment. Oxidative cleavage of
    phosphonothioate and aryl hydrocarbon hydroxylase were elevated at
    10 mg/kg bw. No effect was observed on p-nitroanisole O-demethylase or
    on cytochrome P-450 (Kangsadalampai  et al., 1981).

         Patulin was reported to induce mixed function oxidase in male ICR
    mice treated with 0.5, 1.0 or 2.0 mg/kg bw of patulin intraperitoneally
    (Siraj & Hayes, 1978).

         Patulin inhibited protein prenylation in mouse FM3A cells.
    An inhibition of 50% and 80% was observed at 7 M and 100 M,
    respectively. Protein synthesis, as measured by the incorporation of
    14C-leucine, was also inhibited by patulin. The inhibition was 50%
    at 3 M and >90% at 30 M. In a cell-free assay, patulin inhibited
    rat brain farnesyl protein transferase, one of the enzymes responsible
    for protein prenylation. The inhibition was 50% at a concentration of
    290/M (Miura,  et al., 1992).

         The concentration of glycogen in liver, kidney and intestinal
    tissues was reduced during patulin toxicosis. The decrease in hepatic
    glycogen indicated glucose intolerance which may be due to insulin
    insufficiency. This may be reflected in decreased concentration of
    insulin-dependent enzymes. Glycogen phosphorylase was markedly
    increased, while glycolytic enzymes such as hexokinase and aldolase
    were significantly lowered. Gluconeogenesis was stimulated as
    evidenced by increased glucose-6-phosphatase and fructose-1,6-
    diphosphatase activity (Sakthisekaran  et al., 1989).

     In vitro studies

         Oxygen uptake stimulated by Krebs-cycle intermediates was
    reported to be inhibited in tissue extracts from mice, rats and golden
    hamsters. Inhibition of oxygen uptake in liver homogenates was
    observed at levels of patulin as low as 0.033 mM. Inhibition of oxygen
    uptake in heart and muscle homogenates was greater than in liver

    homogenates. Patulin competitively inhibited succinate dehydrogenase
    in mouse liver homogenates. The P/O ratio was not affected by
    the toxin. In comparative studies, the golden hamster was more
    susceptible, and the rat less susceptible to patulin inhibition than
    the mouse (Hayes, 1977).

         Kidney explants from male Osborne-Mendel rats, when incubated for
    18 h in media containing 0.5, 0.75, or 1.0 mM patulin  in vitro, lost
    their respiratory ability as measured by conversion of 14C-glucose
    to 14CO2. During measurement of respiration, patulin was not
    present in the reaction mixture. At 1.0 mM patulin, respiration was
    increased. Leakage of protein into the medium at a concentration of
    1.0 mM patulin may indicate increased cell membrane permeability
    (Braunberg  et al., 1982).

         Patulin inhibited the  in vitro activity of NaKATPase in
    microsomes prepared from mouse brain. Activity was partially restored
    by washing. Preincubation of patulin with dithiothreitol or
    glutathione prevented the inhibition (Phillips & Hayes, 1978).

         Non-competitive inhibition was demonstrated when patulin was
    incubated with rabbit-muscle aldolase; the Ki was 1.3  10-5 M.
    The cysteine adduct of patulin was a less effective inhibitor (Ashoor
    & Chu, 1973b).

         Patulin, at a level of 4.35 mol/ml, was reported to inhibit by
    29% and 84% the activity of DNA-dependent RNA polymerase I and II
    prepared from rat liver nuclei (Tashiro  et al., 1979).

         Patulin at a level of 200 g/ml inhibited  in vitro the chain
    initiation stage of RNA synthesis in rat liver nuclei (Moule & Hatey,

         Ribonuclease H, prepared from rat liver nuclei, was inhibited by
    patulin  in vitro by 62% at a concentration of 0.32 mol/mol, and by
    47% at a concentration of 1.07 mol/ml (Tashiro  et al., 1979).

         Acid RNAse in human placental microsome and mitochondria-rich
    fractions was increased up to 1.5 times when incubated with
    0.5-3 mg patulin/g of placenta  in vitro (Fuks-Holmberg, 1980).

         Patulin caused a competitive inhibition of lactate dehydrogenase
    from rabbit muscle (K1 = 7.2  10-5 M). The presence of cysteine
    reversed the inhibitory effect of patulin on lactate dehydrogenase
    (Ashoor & Chu, 1973a).

    2.2  Toxicological studies

    2.2.1  Acute toxicity studies

         The acute toxicity of patulin is summarized in Table 1. Toxic
    signs consistently reported in all studies were agitation, in some
    cases convulsions, dyspnea, pulmonary congestion and edema, and
    ulcerations, hyperemia and distension of the GI tract.

         Acute toxicity of i.p. administered patulin was reported to be
    reduced by simultaneous administration of another mycotoxin,
    rubratoxin B (Kangsadalampai  et al. 1981).

         When a patulin/cysteine adduct was administered to mice
    intraperitoneally, no acute toxicity was observed at levels up to
    150 mg of patulin/mouse (Ciegler  et al., 1976).

    2.2.2  Short-term toxicity studies  Mice

         When patulin was administered by gavage in citrate buffer to
    groups of 10 male Swiss ICR mice at doses of 0, 24 or 36 mg/kg bw,
    daily or on alternate days for 14 days, body weight was depressed and
    mortality was increased in a dose-dependent manner. Histopathological
    lesions were found in the GI tract, which included epithelial
    degeneration, haemorrhage, ulceration of gastric mucosa, and exudation
    and epithelial desquamation in the duodenum (McKinley & Carlton,
    1980a).  Rats

         When patulin was administered by gavage to groups of 10 male
    Sprague-Dawley rats at doses of 28 or 41 mg/kg bw, daily or on
    alternate days for 14 days, initial loss of body weight was observed;
    animals recovered after day 4. Mortality was increased in all treated
    groups, but no dose dependency was observed. Gross lesions were found
    in the stomach and small intestine; the gastric mucosa was reddened
    and the stomach was distended. The duodenum and jejunum were distended
    by fluid. Histopathological lesions were found in the stomach which
    consisted of ulceration of the mucosa, epithelial degeneration,
    haemorrhage, and neutrophil and mononuclear cell infiltration
    (McKinley  et al., 1982).

        Table 1.  Acute toxicity of patulin

    Species     Sex         Route       LD50            References
                                        (mg/kg bw)

    Mouse       M           oral        29-48           Escoula, et al., 1977
                                                        Lindroth & von Wright, 1978

                F           oral        46.31           McKinley & Carlton, 1980a
                M&F         "           17              Hayes et al., 1979
                ?           "           25              Katzman et al., 1944

                M           i.p.        5.7-8.17        Ciegler et al., 1976
                                                        Escoula et al., 1977
                                                        McKinley & Carlton, 1980a

                F           i.p.        10.85           Escoula et al., 1977
                M&F         "           7.6             Hayes et al., 1979
                ?           "           4-5.7           Katzman et al., 1944
                                                        Ciegler et al., 1976

                M&F         i.v.        8.57            Escoula et al., 1977
                ?           s.c.        8-10            Katzman et al., 1944
                M           s.c.        10              McKinley & Carlton, 1980a

    Rat         M           oral        30.53-55.0      Escoula et al., 1977
                                                        McKinley et al., 1982

                F           oral        27.79           Escoula et al., 1977
                ?           "           32.5            Dailey et al., 1977b
                M&F         "           108-118         Hayes et al., 1979

                M           i.p.        4.59-100        Escoula et al., 1977
                                                        McKinley et al., 1982

    neonatal    F           oral        5.70            Escoula et al., 1977
    rats        M&F         "           6.8             Hayes et al., 1979

    weanling    M&F         i.p.        5.9             Hayes et al., 1979
    rats        M           i.v.        8.57            Escoula et al., 1977
                M           s.c.        11.0            McKinley et al., 1982
                ?           s.c.        25              Katzman et al., 1944

    Hamster     M           oral        31.5            McKinley & Carlton, 1980b
                            i.p.        10              McKinley & Carlton, 1980b
                            s.c.        23              McKinley & Carlton, 1980b
         Drinking-water containing 0, 25, 85, or 295 mg/litre of patulin
    in 1 mM citrate buffer was given to groups of 6 SPF RIVM:Tox
    (Wistar-derived) rats for 4 weeks. Food and liquid intake were
    recorded three times per week. Body weights were determined at the
    start of the experiment and at termination. Urinalysis, including
    urine volume, bilirubin, and urinary protein were determined in the
    last week. Creatinine clearance was calculated from serum and urine
    levels of creatinine. At termination, the animals were examined
    macroscopically, and the liver, spleen, thyroid glands, brain,
    kidneys, heart, mesenteric lymph nodes, adrenal glands, thymus, testes
    and ovaries were weighed. Histopathological examination was carried
    out on all organs and tissues of the high-dose and the control groups.
    Food and liquid intake were reduced in the mid- and high-dose animals.
    Body weights at the high-dose level were decreased. Creatinine
    clearance was lower in the high-dose animals, but no morphological
    glomerular damage was observed. In the high-dose group, fundic ulcers
    in the stomach were observed in combination with enlarged and active
    pancreatico-duodenal lymph nodes, while villous hyperemia of the
    duodenum was observed at the mid- and high-dose levels. The authors
    suggested, based on normal appearance of the adrenal glands, that the
    observed effects in the GI tract were a direct effect of patulin on
    the tissue, which was not mediated through adrenal gland stimulation
    (stress). The NOEL in this study was 25 mg/litre (Speijers  et al.,
    1985, 1988).

         Albino rats were given 0.1 mg patulin in 0.1 ml propylene glycol
    injected intraperitoneally on alternate days. Control rats were
    treated with propylene glycol only. The animals were sacrificed after
    15 doses. Blood was collected and liver, kidney and intestine were
    used for the estimation of DNA and RNA. In plasma, the total protein
    level, albumin concentration and A/G ratio were significantly
    decreased in the dosed animals. The levels of DNA and RNA in liver,
    kidney and intestine were significantly reduced (Gopalakrishnan &
    Sakthisekaran, 1991).

         Twenty rats/sex received by gavage 0 or 0.1 mg/kg bw patulin
    dissolved in water on alternate days for 30 days. At the end of the
    30th day, rats were killed and the intestine was removed and used for
    the estimation of lipids and Na+ -K+ dependent ATPase.

         Total lipids (25.8%), phospholipid (20.6%) and triglycerides
    (59.8%) decreased significantly whereas total cholesterol levels
    showed a slight increase (12.6%) in the experimental rats. A marked
    inhibition of Na+-K+ dependent ATPase (32.4%) was observed in the
    intestines of experimental rats (Devaraj & Devaraj, 1987).

         Groups of Wistar rats (10/sex/group) were given drinking-water
    containing patulin at concentrations of 0, 6, 30 or 150 mg/litre in
    1 mM citrate buffer for 13 weeks. Food and water intake were decreased
    in the mid- and high-dose groups. Body-weight gain was decreased only
    in animals of the high-dose group. Haematological parameters were
    slightly altered in the high-dose group. No effect of patulin on the
    intestinal microflora was observed. A slight impairment of the kidney
    function and a villous hyperaemia in the duodenum in the mid- and
    high-dose groups were observed. The NOAEL in this study was 6 mg/litre
    drinking-water, equivalent to 0.8 mg patulin/kg bw/day (Speijers,
     et al., 1986).  Hamsters

         When patulin was administered by gavage to groups of 10 male
    Syrian golden hamsters at doses of 0, 16 or 24 mg/kg bw, daily or on
    alternate days for 14 days, loss of body weight was observed and
    mortality was increased in all treated groups, but no dose dependency
    was observed. Gross lesions were found in the stomach and duodenum.
    Histopathological lesions were found in the GI tract and included
    epithelial degeneration, haemorrhage and ulceration (McKinley &
    Carlton, 1980b).  Chickens

         Fifteen one-day old white leghorn chicks were given by intubation
    0 or 100 g patulin, every 48 h. At the end of the 15th dose, the
    birds were fasted overnight and killed. Kidney and intestine were
    removed and assayed. The experimental chicks showed reduced enzyme
    activity. The reduction in the activity of total ATPase in the kidney
    was 40% and in the intestine 52% while the reduction in the Na+-K+
    dependent ATPase activity in the kidney was 46% and in the intestine
    55% (Devaraj  et al., 1986).  Monkeys

         Groups of 1 male and 1 female pigtail monkeys  (Macaca
     nemestrina) received doses of 0, 0.005, 0.05, or 0.5 mg/kg bw/day of
    patulin for 4 weeks. Monkeys of the highest dose group received
    5 mg/kg bw/day patulin for 2 additional weeks. Weekly determinations
    were made of SGOT, SAP, BUN, cholesterol, sodium and potassium as well
    as haematological parameters. Plasma protein electrophoresis was
    performed and glucose and lipoprotein levels were determined. No signs
    of toxicity were noted, except that the monkeys receiving 5 mg/kg
    bw/day of patulin started to reject their food during the last 3 days
    of the experiment. No statistically significant differences were
    observed in any of the parameters studied (Garza  et al., 1977).

    2.2.3  Long-term toxicity/carcinogenicity studies  Rats

         Subcutaneous injections of 0.2 mg of patulin in 0.5 ml of arachis
    oil were administered biweekly for 61 or 64 weeks to 2 groups of 5
    male Wistar rats, weighing 100 g at the start of the experiment. Local
    (fibro)sarcomas was produced at the injection site in 4/4 and 2/4 rats
    surviving at the time when the first tumour was observed. No
    metastases were observed, and of 3 tumours tested only one was
    transplantable in 3 of 12 recipient rats. Control animals receiving
    arachis oil did not develop local tumours (Dickens & Jones, 1961).

         Patulin in water containing 0.05% lactic acid was administered by
    gavage twice weekly to 50 female SPF Sprague-Dawley rats at a dose of
    1 mg/ kg bw for 4 weeks, and 2.5 mg/kg bw for the following 70 weeks
    (total dose: 358 mg/kg bw of patulin). No effects were observed on
    weight gain or survival. No significant differences were observed in
    tumour incidence. The occurrence of 4 forestomach papillomas and 2
    glandular stomach adenomas, as compared to none in the control
    animals, is noteworthy. The Committee noted a discrepancy between the
    reported duration of the study (64 weeks) and the reported duration of
    administration (74 weeks) (Osswald  et al., 1978).

         Groups of 70 FDRL Wistar rats of each sex were exposed to
    0, 0.1, 0.5, or 1.5 mg/kg bw/day of patulin in citrate buffer by
    gavage 3 times/week for 24 months. The rats were derived from the F1
    generation of a 1-generation reproductive toxicity study. Mortality
    was increased in both sexes at the highest dose: all males had died by
    19 months; 19% of females survived until termination at 24 months.
    Body weights of males were reduced at the mid and high dose, but for
    females body weights were comparable in all groups. No difference in
    tumour incidence was observed. The NOEL in this study was 0.1 mg/kg
    bw, administered 3 times weekly (Becci  et al., 1981).

    2.2.4  Reproductive toxicity studies  Rats

         Groups of Sprague-Dawley rats (30/sex/group) received doses of
    0, 1.5, 7.5, or 15.0 mg/kg bw/day of patulin in citrate buffer by
    gavage 5 times per week for 7 weeks before mating. The pregnant dams
    were gavaged daily at the same levels during gestation. Half the
    dams were sacrificed on day 20 of gestation, and used for teratological
    evaluation. The remaining dams were allowed to produce the F1
    generation. Some of the F0 and F1 males were used for a dominant

    lethal experiment. Twenty three controls and 15 low-dose animals per
    sex were continued to produce an F2 generation. One-half of the
    latter generation were again used for teratological evaluation.
    Haematological and blood chemistry examinations were performed on
    10 males and 10 females of the F2 generation 23 days after
    weaning. The only lesion found at necropsy of parent animals was
    gaseous distension of the GI tract. All treated males of the F0
    generation had a dose-related reduction in weight gain. High mortality
    occurred at 7.5 and 15.0 mg/kg bw/day in both males and dams. Although
    Jitter size at 7.5 mg/kg bw/day was comparable to controls, survival
    of male progeny was severely impaired. At the 1.5 mg/kg bw/day level,
    pup growth of both sexes was reduced, and there was increased
    mortality among the F2 females. No significant alterations were
    found in the haematology and blood chemistry levels in selected
    animals of the F2 generation (Dailey  et al., 1977b).

         Groups of FDRL Wistar rats (50/sex/group) were exposed to
    0, 0.1, 0.5, or 1.5 mg/kg bw/day of patulin in citrate buffer by
    gavage for 4 weeks before mating, and pregnant females were dosed
    through gestation and lactation. The parent generation was sacrificed
    after weaning. Body-weight gain was comparable among groups. Ten
    females died in the high-dose group. Reproductive parameters such as
    mating success, litter size, fertility, gestation, viability, and
    lactation indices, and pup weight at birth, at 4 days and at weaning,
    were not statistically different among experimental groups.
    Histopathological evaluation of grossly abnormal tissues of the F0
    generation did not show any effects of patulin treatment. The F1
    generation was used for a 2-year toxicity/carcinogenicity study (see
    section (Becci  et al., 1981).

    2.2.5  Special studies on antibiotic activity

         Twelve species of bacteria and two species of yeast were tested
    for sensitivity against 11 different mycotoxins, including patulin,
    using a disc diffusion assay.  Bacillus brevis appeared to be the
    most sensitive microorganism. The lowest amount that could be detected
    under optimal conditions was 1 g/disc for patulin (Madhyastha
     et al., 1994).

         Clear synergy was shown with patulin plus rifampin and patulin
    plus bottromycin. Synergy of patulin with efrotomycin was weak and
    there was no synergy of patulin plus kasugamycin (Dulaney & Jacobsen,

    2.2.6  Special studies on antitumour activity

         A comparison was made between the cytotoxicity and antitumour
    activity of patulin and five structural analogs (isopatulin,
    dehydroisopatulin, dimethylisopatulin, trimethylisopatulin and
    isopropylisopatulin).  In vitro assays using L1210 and P 388 cells
    showed that the structure of the pyranic ring as well as the nature of
    the substituents influenced the observed activities. Among the five
    structural analogs of patulin assayed  in vivo against Ehrlich
    carcinoma, L1210 and P388 leukemias, dehydroisopatulin was the only
    one being active on all 3 types of tumours at a dose of 100 mg/kg
    bw/day. The ratios between the LD50 in mice and the active dose was
    5, while with patulin it was 10 (Seigle-Murandi  et al., 1992).

    2.2.7  Special studies on cytotoxicity

         The ID50 (inhibitory dose) of patulin tested on the protozoan
     Tetrahymena pyriformis was 0.32 g/ml (Nishie,  et al., 1989).

         Patulin at a concentration of 3.2 g/ml inhibited by 50% the
    growth rate of the ciliate  Tetrahymena (Brger  et al., 1988).

         To evaluate its inhibitory effect on cells, hepatoma tissue
    culture cells in suspension were incubated in the presence of 30 M of
    patulin for 7 h and investigated by transmission and scanning electron
    microscopy. The most significant difference observed between treated
    and control cells was the disorganization of the cytoplasmic
    microfilaments in the treated cells (Rihn,  et al., 1986).

         In an immortalized rat granulosa cell line, effects of patulin on
    GSH levels and alterations in the partitioning of rhodamine 123 were
    detected at 0.1 M within 1 h. Alterations in Ca2+ homeostasis,
    intracellular pH and gap junction mediated intercellular communication
    were detected between 1 and 2 h with 1.0/M patulin (Burghardt  et al.,

         A study in cultural renal cells on the effect of patulin on ion
    influx, and the influence of dithiothreitol and glutathione on patulin
    effects was performed. It was hypothesized that patulin altered
    intracellular ion content via Na+-K+ ATPase and non-Na+-K+ ATPase
    mechanism (Hinton  et al., 1989).

         In cultured renal cells LLC-PK1, concentrations of
    patulin above 10/M caused a transient increase in intracellular
    electronegativity (< 1 h) followed by a sustained depolarization
    (> 1 h), which was correlated with complete Na+ influx, K+
    efflux, total LDH release, and bleb formation. However, patulin
    concentrations of 5-10 M caused a sustained increased intracellular
    electronegativity (4-8 h) which was associated with partial Na+
    influx and K+ efflux, no significant LDH release, and relatively few
    blebs. The hyperpolarizing effect may be a result of increased
    intracellular electronegativity. The toxic effects of patulin are
    irreversible in LLC-PK1 cells, even after short pretreatment with
    patulin (Riley  et al., 1990).

         In LLC-PK1, cells exposed to 50 M patulin lipid peroxidation,
    abrupt calcium influx, extensive blebbing and total LDH release
    appeared to be serially connected events with each representing a step
    in the loss of structural integrity of the plasma membrane. Patulin
    also caused depletion of nonprotein sulfhydryls, increased 86Rb+
    efflux, dome collapse and eventually the loss of cell viability (Riley
    & Showker, 1991).

    2.2.8  Special studies on genotoxicity

         The results of  in vitro and  in vivo genotoxicity studies with
    patulin are summarized in Tables 2 and 3, respectively.

         Patulin was negative in mutagenicity tests with  S. typhimurium
    but was clearly positive in the initiator tRNA acceptance assay for
    carcinogens (Hradec & Vesely, 1989).

         The mutagenic effect of patulin was studied with a mutant of
    bacteriophage M13am6H1. A 50% decrease in liberation of M13 phage per
    cell (ED50) was observed at a concentration of 0.85 g patulin/ml,
    and a 50% decrease in growth rate of  E. coli host cells was observed
    at a much higher concentration of the mycotoxin (6.3 g/ml). The
    reversion frequency of M13am6H1 to the wild-type phenotype in the
    presence of patulin compared to the spontaneous reversion increased by
    a factor of 7-19.5 depending on the addition of patulin to bacteria or
    phages only or simultaneously.

         The same authors studied the effects of patulin on protein and
    DNA synthesis. At a concentration of 3.2 g/ml, the protein synthesis
    of the ciliate  Tetrahymena was inhibited by 85% and the RNA
    synthesis by 86% compared with the control. Four hours after addition
    of patulin, DNA synthesis was reduced to 20%; it rose to the value of
    the control after an additional 2 h. An  in vitro system could be
    developed consisting of permeabilized cells of  Tetrahymena. This
    system allowed the separation of regulatory and secondary effects
    induced by patulin. Patulin reduced DNA synthesis by 50%, whereas RNA
    and protein synthesis were less inhibited than in the  in vivo system
    (Burger  et al., 1988).

        Table 2.  Results of in vitro genotoxicity assays on Patulin

    Test system           Test object            Concentration          Result        Reference

    Ames test             E. coli                1 g/ml (to phage)     Positive      Burger et al., 1988
                          M 13am6H1              &/or 5 g/ml
                                                 (to bacteria)

    Ames test(3,4)        S. typhimurium         0.01, 0.1, 1,          Negative      Ueno et al., 1978
                          TA98, TA100            10, 100, & 500

    Ames test(3)          S typhimurium          0.25 23, 25 &          Negative      Wehner et al., 1978
                          TA98, TA100,           250 g/plate
                          TA1535, TA1537

    Ames test(3)          S, typhimurium         0.1, 1, 10 & 100       Negative      Kuczek et al,, 1978
                          TA1535, TA1537,        g/plate

    Ames test(5)          S. typhimurium         5, 10, 20 & 30         Negative      Von Wright & Lindroth,
                          TA98, TA100            g/plate                             1978

    Ames test(3,5)        S. typhimurium         <0.0065                Negative      Bartsch et al., 1980
                          TA100, TA1538          moles/plate

    Ames test(5)          S. typhimurium         12-960 g/plate        Negative      Wurgler et al., 1991
                          TA102                  Conc. >35
                                                 were toxic;

    Table 2 (cont'd)

    Test system           Test object            Concentration          Result        Reference

    Chromotest(3)         E. coli                0.01, 0.02 & 0.05      Positive      Auffray &
                          K12 PQ37               /ml                   (No S-9)      Boutibonnes, 1987
                                                                        (with S-9)

    SOS Chromotest        E. coli K12            0.01, 0.02 & 0.05      Weakly        Auffray &
                                                 g/l                   positive      Boutibonnes, 1988

    Chromotest(2)         E. coli PQ37           0.001-30 g/ml         Negative      Krivobok et al., 1987

    Recombinogenesis      B. subtilis            20 & 100 g/disc       Positive      Ueno & Kubota, 1976

    Prophage induction    E. coli                5, 10, 25 & 50         Positive      Lee & Roschenthaler,
                          X8011 (lambda)         g/ml                                1986

    Spot test             E. coli K12            1-10 g/assay(2)       Positive      Auffray &
                                                                                      Boutibonnes 1986

    Reverse               S. cerevisiae          50 /No S-9) &          Positive      Kuczuk et al., 1978
    mutagenesis(3)        D-3                    100 (with S-9)

    Forward               S. cerevisiae          10, 25, 50 & 75        Positive      Mayer & Legator
    mutagenesis           (haploid)              g/ml                                1969

    Table 2 (cont'd)

    Test system           Test object            Concentration          Result        Reference

    Forward               FM3A mouse             0.032, 0.1, &          Positive      Umeda et al., 1977
    mutagenesis           mammary                0.32 g/ml
    (8-azoquanine         carcinoma cells

    SOS microplate        E. coli PQ37           2, 8, 24 g/ml         Negative      Sakai et al.. 1992

    Somatic mutations     Drosophila             3.210-2,              Weakly        Belitsky et al., 1985
                          melanogaster           3.210-3, M            positive

    Chromosome            FM3A mouse             0.032, 0.1 & 0.32      Positive      Mori et al., 1984
    aberration            mammary                g/ml
    induction             carcinoma

    Chromosome            Chinese hamster        1, 2.5. 5 & 10 M      Positive      Thust et al., 1982
    aberration            V79-E cells                                   (No S-9)
    induction(3)                                                        Negative

    Chromosome            Human                  3.5 M                 Positive      Withers, 1966
    aberration            leucocytes

    Cell cycle            Prim. Chinese          0.5, 1 & 2 g/ml       Positive      Kubiak &
    retardation           hamster cells                                               Kosz-Vnenchak, 1983

    Table 2 (cont'd)

    Test system           Test object            Concentration          Result        Reference

    Cell cycle            Human                  0.075 & 0.30           Positive      Cooray et al., 1982
    retardation           peripheral blood       mg/ml

    Cell cycle            Human                  0.075 & 0.30           Positive      Cooray et al., 1982
    retardation           peripheral blood       g/ml

    Sister chromatid      Chinese hamster        1, 2.5, 5 & 10 M      Negative      Thust et al., 1982
    exchange              V79-E cells

    Sister chromatid      Prim. Chinese          0.5, 1 & 2 mg/ml       Positive      Kubiak & Vnencha, 1983
    exchange induction    hamster cells

    Sister chromatid      Human                  0.075, 0.10, 0.20      Weakly        Cooray et al., 1982
    exchange induction    peripheral blood       & 0.30 mg/ml           positive

    DNA synthesis         T. pyriformis          3.2 g/ml              Positive      Burger et al., 1988

    DNA synthesis         AWRF cells             1, 2, 4 & 8 g/ml      Positive      Stetina & Votova, 1986
    retardation           CHO cells              0.24, 0.5, 1, 2, 4

    Table 2 (cont'd)

    Test system           Test object            Concentration          Result        Reference

    DNA breakage          ColE1 plasmid          0.25, 0.5, 1.0 &       Negative      Lee & Roschenthaler,
                                                 5.0 mM(1)                            1986
                          Lambda DNA             0.5. 1, 5, 10 &
                                                 14 mM

    Unscheduled DNA       Primary ACI rat        60 & 600 M            Negative      Mori et al., 1984
    synthesis induction   hepatocytes

                          Primary C3H            65 & 650 M            Negative      Mori et al., 1984

    DNA breakage          E. coli                10, 20, 25 & 50        Positive      Lee & Roschenthaler,
                          D110 polA              g/ml                                1986

    DNA-repair            human or rat           1.610-3 -             Negative      Belitsky et al., 1985
                          liver cells            1.610-3

    DNA breakage          FM3A mouse             1.0, 3.2, 10 g/ml     Positive      Umeda et al., 1977
                          carcinoma cells

    DNA breakage          AWRF cells             2 & 10 g/ml           Positive      Stetina & Votava, 1986
                          CHO cells              2, 8 & 10 g/ml

    DNA synthesis         human                  1.0105                Negative      Yanagisawa et al., 1987
    inhibition test       fibroblasts

    Table 3.  Results of in vivo genotoxicity assays on Patulin

    Test system        Test object           Concentration          Result       Reference

    Chromosome         Chinese hamster       oral 2  10 and 20     Positive     Roll et al., 1990
    aberrations        bone marrow           mg/kg bw

    Chromosome         Chinese hamster       2  20 mg/kg bw        Positive     Korted et al., 1979
    aberration         bone marrow cells     by gavage(6)

    Chromosome         Chinese hamster       2  1, 10 & 20         Positive     Korte, 1980
    aberration         bone marrow cells     mg/kg bw by
    induction                                gavage

    Chromosome         Chinese hamster       2  10 & 20 mg/kg      Positive     Korte & Ruckert, 1980
    aberration         bone marrow cells     bw

    Host mediated      S. typhimurium        i.m. 3  <500 g       Negative     Gabridge & Legator, 1969
    assay in Swiss     G46
    albino mice

    Host mediated      S. typhimurium        10  20 mg/kg bw       Negative     Von Wright & Lindroth,
    assay in male      TA1950, TA1951        gavage                              1978
    NMRI mice

    Dominant lethal    ICR/Ha Swiss          0.1 & 0.3 mg/kg        Negative     Epstein et al., 1972

    Dominant lethal    Sprague-Dawley        1.5 mg/kg bw           Negative     Dailey et al., 1977b
    assay              rats                  5x/wk  10-11 wk,
                                             by gavage

    Table 3.  (cont'd)

    Test system        Test object           Concentration          Result       Reference

    Dominant lethal    Texas ICR x           3.0 mg/kg bw i.p.      Negative     Reddy et al., 1978
    assay              Sprague-Dawley

    Sister chromatid   Chinese hamster       2  1, 10 & 20         Negative     Korte, 1980
    exchange           bone marrow cells     mg/kg bw by
    induction                                gavage

    Dominant lethal    NMRI mice (m)         i.p 2.5 and 5          Negative     Roll et al., 1990
    assay                                    mg/kg bw

    (1)    Positive when CuCl2 & NADPH were added
    (2)    Both with and without S-9 fraction (source not specified)
    (3)    Both with and without rat liver S-9 fraction
    (4)    Both with regular plate and preincubation methods
    (5)    Both with and without mouse liver S-9 fraction
    (6)    Effect negative if animals first given ethanol as only liquid for 9 wk prior to exposure
    2.2.9  Special studies on immunotoxicity  In vitro studies

         Peritoneal exudate cells of mice (C57BL/6J) collected by washing
    the peritoneal cavity, were preincubated for 2 h with 0.01-2 g
    patulin/ml. Phagocytosis and phagosome-lysosome fusion were diminished
    above 0.1 g/ ml, and lysosomal enzymes and microbiological activity
    above 0.5 g/ml, whereas O2 production was inhibited only above
    2 g/ml (Bourdiol  et al., 1990).

         The effects of patulin were investigated on immunological
    responses of Balb/c mice.  In vitro, patulin had a stimulatory effect
    on splenocytes at lower concentration (1 nM to 10 nM) and strongly
    inhibited lymphocyte proliferation at higher concentrations (ID50
    from 0.02 to 0.24/uM depending on mitogens) (Pancod  et al., 1990).

         At concentrations from 0.25-1 g/ml, patulin decreased the
    chemotactic index of dog neutrophilic granulocytes stimulated by
    opsonized zymosan. At the same concentrations patulin favoured the
    migration of the cells. At 1 g/ml it inhibited the liberation of
    superoxide ions by neutrophils, but did not modify their ability to
    phagocyte  Saccharomyces cerevisiae even at concentrations up to 10
    g/ml. The immunosuppressive actions may be explained by a fixation of
    patulin on sulfhydric groups present on the neutrophil membrane
    (Dubech,  et al., 1993).

         In alveolar macrophage harvested from male Long-Evans hooded
    rats, patulin caused a significant increase in mean cell volume after
    2 h exposure at 10-3 M. Chromium release from alveolar macrophage
    following exposure to patulin was both time- and concentration-
    dependent. Treatment with > 1.5  10-4 M caused significant
    chromium release within 30 minutes. ATP concentrations in alveolar
    macrophage monolayer cultures were markedly inhibited within 1 h at
    concentrations > 5  10-5 M patulin. Incorporation of [3H]-precursors
    into protein and RNA was also strongly inhibited by patulin. Inhibition
    was both time- and concentration-dependent for both classes of molecules
    but protein synthesis was sensitive to 10- to 100- fold lower
    concentrations of patulin than RNA synthesis at the same time interval.
    The dose producing 50% inhibition at 1 h (ED50) was estimated at
    ca. 1.6  10-6 M and 1.0  10-5 M for [3H]-leucine and [3H]-uridine
    incorporation, i.e. protein and RNA synthesis, respectively. Patulin
    strongly inhibited phagocytosis of [51Cr]-sheep erythrocytes and
    there was significant inhibition of phagocytosis at >5  10-7 M
    patulin (Sorenson  et al., 1986).  In vivo studies

         The effects of patulin were investigated on immunological
    responses of Balb/c mice. In mice, patulin at dose levels of 2 and
    4 mg/kg bw significantly reduced delayed type hypersensitivity to
     Bordetella pertussis antigen and did not reduced anti-KLH antibody
    production (Paucod  et al., 1990).

         Mice (Swiss female IFA CREDO) receiving 10 mg/kg bw/day patulin
    for 4 days showed enhanced resistance to i.p. challenge with 108
    viable  Candida albicans at day 2. Immunoglobulin levels (IgA, IgM
    and IgG) were markedly depressed (10-75%) (Escoula  et al., 1988a).

         Mice (Swiss female IFA CREDO) were given by gavage 10 mg/kg
    bw patulin daily from day 0 to day 4, and rabbits received
    intraperitoneally 2.5 mg/kg bw. The mice were lymphopenic on days
    5 and 10, but not on day 20.

         There was no effect on neutrophil count on day 5. A significant
    suppression of the chemilumminescene response of peritoneal leucocytes
    was observed in both species. Mitogenic responses of mice splenic
    lymphocytes and rabbit peripheral cells were slightly suppressed
    (ConA) by treatment with 0.05 g/ml and markedly inhibited with
    0.5 g/ml. The inhibition was more pronounced on B-cell mitogen
    compared with T-cell mitogen. In mice and rabbits IgG, IgA and IgM
    levels obtained on day 5 were lower when treated with patulin (Escoula,
     et al., 1988b).

         Patulin inhibited DNA synthesis in peripheral lymphocytes. These
    effects were mitigated by cysteine which suggested that sulfhydryl
    binding was involved in patulin induced toxicity. In mice, an
    increased resistance to  Candida albicans was observed and decreased
    concentrations of circulating immunoglobulin. In rabbits decreased
    serum immunoglobulin, reduced blasto-genesis of lymphocytes and
    reduced chemiluminescence of peritoneal leucocytes were observed. No
    details about concentrations were given (Sharma, 1993).

    2.2.10  Special studies on embryotoxicity

         Groups of 25 albino rats (sex not specified) weighing 25-30 g
    received 0 or 100 mg of patulin in propylene glycol intraperitoneally
    on alternate days (dose approximately 1.6 mg/kg bw/day) for 1 month.
    The patulin-treated animals showed convulsions, tremors, impaired
    locomotion, stiffness of hindlimbs, and wagging of the head. Patulin
    inhibited acetylcholinesterase and NaKATPase in the cerebral
    hemisphere, cerebellum and medulla oblongata. Concomitantly,
    acetylcholine levels were raised in these brain segments (Devaraj
     et al., 1982a).

    2.2.11  Special studies on teratogenicity and embryotoxicity  Mice

         Twelve pregnant Swiss mice received 0 or 2 mg/kg bw/day of
    patulin in water containing 0.05% lactic acid twice daily by gavage
    for 6 days starting 14 days after mating. The control mice received
    0.05% lactic acid by gavage.

         Mean survival time was significantly reduced in the patulin
    treated dams, while 2/12 control animals and 5/12 experimental animals
    developed tumours. Of the offspring, 8/43 male and 11/52 female
    suckling mice died in the first 6 days of life, with hyperemia and
    bleeding in the brain, lungs and skin. When these early deaths were
    excluded from the calculations, patulin did not affect survival time
    in the animals exposed  in utero. No evidence of carcinogenicity was
    observed in the offspring exposed only to patulin  in utero (Osswald
     et al., 1978).

         Groups of 22-31 mice (NMRI) received orally during days 12
    and 13 of gestation 0 or 3.8 mg/kg bw/day, or i.p. 0, 1.3, 2.5 or
    3.8 mg/kg bw/day. Higher dose levels were maternally toxic. Oral
    administration caused no effects on the number of implantation,
    delivered fetuses, number of resorption, dead fetuses, fetal weight,
    or malformations of the skeleton and organs. Intraperitoneal
    administration showed at 3.8 mg/kg bw/day a slight increase in early
    resorption, compared to controls. An increase in cleft palate was seen
    (10.6% compared to 1.5% in controls), and an increase in malformations
    of the kidney (2.8% compared to none in the control group) were also
    seen at this dose level (Roll  et al., 1990).  Rats

         In a 2-generation reproductive toxicity study (see section, offspring of 15 Sprague-Dawley dams of the F1 and F2
    generation exposed by gavage to 0 or 1.5 mg/kg bw/day of patulin in
    citrate buffer were evaluated for teratological abnormalities. Patulin
    caused an increase in resorption in the F1 litters, but this effect
    was not observed in the F2 generation. The average weight of male
    fetuses of the F2 generation was significantly less than controls.
    No increase in skeletal or soft tissue abnormalities was observed
    (Dailey  et al., 1977b).

         However, when patulin was administered i.p. to groups of 10-17
    pregnant Charles River CDI rats at doses of 1.5 or 2.0 mg/kg bw/day, a
    significant decrease in average fetal body weight was observed at the
    lower dose, and at 2.0 mg/kg bw/day all implanted embryos were
    resorbed (Reddy,  et al., 1978).  Chickens

         Patulin was injected into the air cell of chick eggs. It was
    reported to be embryotoxic at levels of 2.4-69 g/egg depending on the
    age of the embryo, and teratogenic at levels of 1-2 g/egg. Patulin/
    cysteine adducts exhibited the same toxic effects, but at much higher
    doses: 15-150 g of patulin equivalents (Ciegler  et al., 1976).  In vitro studies

         Whole rat embryo culture was used to determine the teratogenic
    potential of patulin  in vitro. Embryos were exposed to untreated or
    patulin-treated (0 - 62 M) rat serum for 45 h. The embryos exposed to
    62 M patulin were not evaluated because they did not survive the 40 h
    incubation time. The results indicated that patulin induced a
    concentration-dependent reduction in protein and DNA content, yolk sac
    diameter, crown rump length, and somite number count. Patulin
    treatment also resulted in an increase in the frequency of defective
    embryos. Anomalies included growth retardation, hypoplasia of the
    mesencephalon and telencephalon, hyperplasia and/or blisters of the
    mandibular precess (Small  et al., 1992, summary only).

    2.3  Observations in humans

         Patulin was tested as an antibiotic for treatment of the common
    cold in humans. Application was through the nasal route (1:10 000 or
    1:20 000 solutions, every 4 h). Most of the information is anecdotal
    (Gye, 1943).

         A report on a controlled trial failed to identify the number of
    patients tested, and was unclear as to which clinical tests were
    performed to support the authors assertion that no ill effects were
    observed (Hopkins, 1943).


         In rats, most of the administered dose was eliminated within 48 h
    in faeces and urine, less than 2% being expired as carbon dioxide. No
    other metabolites have been identified. About 2% of the administered
    dose was still present after 7 days, located mainly in erythrocytes.

         Patulin has a strong affinity for sulfhydryl groups, which
    explains why it inhibits the activity of many enzymes. Patulin adducts
    formed with cysteine were less toxic than the unmodified compound in
    acute toxicity, teratogenicity, and mutagenicity studies.

         In acute and short-term studies, patulin caused gastrointestinal
    hyperaemia, distension, haemorrhage and ulceration. Pigtail monkeys
     (Macaca nemestrina) tolerated patulin consumption of up to 0.5 mg/kg
    bw/day for 4 weeks without adverse effects.

         The NOEL in a 13-week toxicity study performed in rats was
    0.8 mg/kg bw/day, based on a slight impairment of kidney function and
    a villous hyperaemia in the duodenum in the mid- and high-dose groups.

         Two reproductive toxicity studies in rats and teratogenicity
    studies in mice and rats were available. No reproductive or
    teratogenic effects were noted in mice or rats at dose levels of up
    to 1.5 mg/kg bw/day. However, maternal toxicity and an increase in
    the frequency of fetal resorptions were observed at higher levels,
    which indicated that patulin was embryotoxic.

         Both  in vitro and  in vivo experiments indicated that patulin
    had immuno-suppressive properties. However, the dose levels at which
    these effects occurred were higher than the NOEL in both the
    short-term toxicity study and a combined reproductive toxicity/
    long-term toxicity/carcinogenicity study.

         Although the data on genotoxicity were variable, most assays
    carried out with mammalian cells were positive while assays with
    bacteria were mainly negative. In addition, some studies indicated
    that patulin impaired DNA synthesis. These genotoxic effects might be
    related to its ability to react with sulfhydryl groups and thereby
    inhibit enzymes involved in the replication of genetic material.
    Nevertheless, it was concluded from the available data that patulin is

         The mortality seen in short-term toxicity, reproductive toxicity
    and long-term toxicity studies with conventional rats, due to
    dilatation of the gut and/or pneumonia, was most probably secondary to
    the fact that patulin acts like an antibiotic on Gram-positive
    bacteria, thereby giving a selective advantage to pathogenic
    Gram-negative bacteria. This conclusion was supported by the fact
    that, in 13-week studies at similar dose levels with specific
    pathogen-free (SPF) rats, no such mortality was seen.

         In combined reproductive toxicity, long-term toxicity/
    carcinogenicity study in rats, a dose level of 0.1 mg/kg bw/day of
    patulin produced no effect in terms of decreased weight gain in males.
    However, as patulin was administered only three times per week during
    24 months, the NOEL derived from this study was 43 g/kg bw/day.

         An additional long-term carcinogenicity study in a rodent species
    other than the rat, which was recommended at the previous meeting for
    the further evaluation of the toxicity of patulin, was not available.


         Since in the most sensitive experiment, patulin was administered
    only three times per week, the existing PTWI was changed. As it does
    not accumulate in the body and in the light of the consumption
    pattern, the PTWI was changed to a provisional maximum tolerable daily
    intake (PMTDI). Based on a NOEL of 43 g/kg bw/day and a safety factor
    of 100, a PMTDI of 0.4 g/kg bw was established.

         Submission of the results of a long-term toxicity/carcinogenicity
    study in a rodent species other than the rat is desirable.

         Patulin levels in apple juice are generally below 50 g/litre and
    maximum intakes have been estimated to be 0.2 g/kg bw/day for
    children and 0.1 g/kg bw/day for adults, i.e. well below the
    tolerable intake established by the Committee. However, apple juice
    can occasionally be heavily contaminated and continuing efforts are
    therefore needed to minimize exposure to this mycotoxin by avoiding
    the use of rotten or mouldy fruit.


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    See Also:
       Toxicological Abbreviations
       Patulin (WHO Food Additives Series 26)
       PATULIN (JECFA Evaluation)
       Patulin (IARC Summary & Evaluation, Volume 10, 1976)
       Patulin (IARC Summary & Evaluation, Volume 40, 1986)