Penicillium verrucosum in feed of ochratoxin A positive swine herds

Ochratoxin A contamination of cereal feed grain was monitored during October 1989–September 1990 by analysis of blood samples from slaughter swine in Sweden. The detection of ochratoxin A in swine blood was used as a method to identify swine herds fed ochratoxin A contaminated feed. The contamination level of ochratoxin A in the blood of the positive herds was in the range 2–45 ng/ml with the mean concentration 5.2 ng/ml. Feed samples for mycological analysis were collected from both ochratoxin A positive herds (2 ng/ml blood) and ochratoxin A negative herds (<2 ng/ml blood). From the ochratoxin A positive herds and the ochratoxin A negative herds 22 and 21 feed samples were collected, respectively. No quantitative differences in mould content, as determined by colony forming units, were observed between the two groups. However, there were differences in the mycoflora. The incidence of storage fungi (Penicillium and Aspergillus spp.) was significantly higher (p < 0.05) in feed from ochratoxin A positive herds. Particularly, Penicillium verrucosum was found to be significantly more common (p < 0.001). Altogether 274 isolates were screened for their ability to produce ochratoxin A. Ochratoxin A producers were found only within P. verrucosum; 38% of the 63 isolates produced detectable amounts of ochratoxin A. Ochratoxin A producing isolates of P. verrucosum were found in 60% of the feed samples collected from ochratoxin A positive swine herds and in one sample (5% ) of the feed samples collected from the ochratoxin A negative herds.     

Citrinin,ochratoxin A and iron. Possible implications for their biological function and induction of nephropathy

Experiments with Neisseria meningitidis have shown that Fe³⁺ to some extent can reverse the toxicity of ochratoxin A and citrinin, as measured by inhibition zones around impregnated paper discs. Similar phenomena were observed with the less toxic ochratoxin B. Zearalenone also inhibited growth, but its effect was not counteracted by iron. The mycotoxins aflatoxin B₁ and deoxynivalenol did not inhibit bacterial growth at all. Desferal (deferoxamine) also inhibited growth of meningococci, but iron totally abolished this inhibition. The results indicate that ochratoxin A and citrinin interfere with iron metabolism in this organism but that other additional toxic mechanisms are involved as well since a marked growth inhibition by both toxins was also observed in the presence of iron. One function of ochratoxin A and citrinin in nature could consequently be to affect the iron uptake of other competing microorgansms.Since both toxins interfere with iron and both cause nephropathy, a possible connection between these properties and lipid peroxidation is also briefly discussed.Abbreviations DON deoxynivalenol - OA ochratoxin A - OB ochratoxin B     

Assessment of ochratoxin A and tenuazonic acid in Canadian ice-wines

Twenty-six samples of commercial ice-wine made from late-harvested grapes were assayed for the mycotoxins ochratoxin A and tenuazonic acid. Canadian wines originated in British Columbia (18) and Ontario (8). For comparison two German wines from Hesse were also studied. Four additional samples of research ice-wine originating in were also studied. In all wine samples, assays using immuno-affinity chromatography and fluorescence liquid chromatography indicated ochratoxin A below 0.15 μg/L, the limit of determination of the method. Tenuazonic acid was determined by solidphase micro-extraction and liquid chromatography and was below the limit of determination (70 μg/L) in all samples. The European Union food tolerance limit for ochratoxin A in wine is 2 μg/L. A tolerance for tenuazonic acid has not yet been established.     

Biodegradation of ochratoxin A by Aspergillus section Nigri species isolated from French grapes: a potential means of ochratoxin A decontamination in grape juices and musts

Ochratoxin A (OTA) is a very dangerous mycotoxin, the presence of which is often reported in different foods, as well as in beverages such as grapes, grape juices and wines. Detoxifying these products is therefore of prime importance in protecting consumer health, and biological approaches have been the most promising methods. In this report, 40 isolates representing the black apergilli species Aspergillus carbonarius, A. niger aggregate and A. japonicus, isolated on French grapes, were assessed for OTA degradation capacities in CZAPEK yeast extract broth (CYB) and in a synthetic grape juice medium (SGM) contaminated with OTA at 2 mg L(-1) (5 microM). It was clearly observed that in both media these fungi had the ability to degrade OTA to OTalpha (ochratoxinalpha). However, there were differences between the media used and species tested during OTA degradation. In SGM and CYB, 77% and 45% of the isolates, respectively were able to degrade more than 80% of the OTA. Despite a better growth on SGM, specific OTA degradation was higher on CYB for most of the isolates. Kinetic studies carried out on SGM with three black Aspergillus isolates all showed different OTA degradation rates. After 9 days of incubation, OTalpha had decreased, whereas an unknown compound appeared. A. niger could be the first interesting species for OTA detoxification processes, followed by A. japonicus.     

半胱胺对鹅胰液分泌及胰酶活性的影响

目的 :研究半胱胺对鹅胰液分泌及胰酶活性的影响。方法 :12只装有胰腺～十二指肠长久性瘘管的成年鹅。试验采用自身对照 ,试验期在日粮中一次性添加半胱胺 (10 0mg/kgbw)。连续收集计量胰液并测定胰酶活性。 结果 :①半胱胺使鹅胰液的分泌速率较对照期显著上升 (2 4 0 .16 % ,P <0 .0 1) ,其中白天升高 2 34.4 5 % ,夜间增高了2 5 3.70 %。②试验期单位容积胰蛋白酶的活性较对照期升高了 4 9.0 5 % (P <0 .0 1) ,而胰脂肪酶和胰淀粉酶的活性却较对照期分别降低了 2 5 .4 4 %和 2 1.95 % ,且变化幅度具有昼夜的差别。③试验期胰腺每小时分泌胰蛋白酶、胰脂肪酶和胰淀粉酶的总活性较对照期分别升高 4 0 6 .88% (P <0 .0 1)、15 3.5 8% (P <0 .0 1)和 16 6 .5 9% (P<0 .0 1) ,白天增加的幅度较夜晚大。结论 :半胱胺能促进鹅胰液的分泌 ,增加胰液中蛋白酶、脂肪酶和淀粉酶分泌的总量 ,从而提高鹅对饲料的消化能力 ,适应机体生长对营养的需求     

Occurrence of deoxynivalenol (DON) and ochratoxin A (OTA) in dog foods

The commercially available dog food samples (29 dry foods and 11 wet foods) were analysed for deoxynivalenol (DON) and ochratoxin A (OTA) using ELISA. All (100%) dry foods were contaminated with DON with various amount of the toxin (22-1837 μg/kg). In wet food 3 samples were found to be positive for DON in the range of 95-170 μg/kg. There were a few samples contaminated with OTA: 3 samples in dry foods (7-40 μg/kg) and 2 samples in wet foods (45 and 115 μg/kg).     

Uptake and genotoxic effects of ochratoxin A in cultured porcine urinary bladder epithelial cells

Uptake of radiolabelled ochratoxin A (OTA) into porcine urinary bladder epithelial cells (PUBEC) was measured at neutral (pH 7.5) or acidic (pH 5.0) conditions. Genotoxicity of OTA was evaluated with the Comet assay and cytotoxicity with the neutral red uptake assay. At acidic pH-conditions, the bladder cells were able to take up more OTA than at neutral conditions. Cytotoxic effects were not increased at pH 5.0 compared to pH 7.5, but higher OTA uptake correlated with stronger genotoxic effects in the Comet assay at pH 5.0 compared to pH 7.5. These results demonstrate that uptake of OTA has to be regarded as an important factor for the toxicity of OTA as adverse effects depend on the amount of OTA taken up by the cells. Presented at the 25^th Mykotoxin Workshop in Giessen, Germany, May 19–21, 2003     

Explaining co-occurrence among helminth species of lesser snow geese (Chen caerulescens) during their winter and spring migration

The digestive tracts of 771 lesser snow geese (Chen caerulescens) collected from January to May 1983 from 12 locations (27 samples) were examined for helminth parasites to determine whether parasite species present in wintering geese or in spring migrants occurred independently of each other. Nine helminth species were identified. Seven had mean prevalences >5% and were the focus of this study. Six of those species were waterfowl generalists, one was a goose specialist. Our primary objective was to assess the potential contribution of factors, other than species interactions, in determining patterns of co-occurrence between helminth species. There were few negative relationships between helminth species, regardless of whether presence-absence or abundance data were used. However, some species pairs showed recurrent and significant co-occurrences. There were similar and significant effects of timing of sampling, host gender, and host age, on prevalence and mean abundance of particular species. Co-occurrences were found for those species that showed seasonal declines in prevalence, for those expected to have high colonizing ability based on host age profiles (using abundance data), and for abundant species that may have shared vectors or environmental conditions favorable for transmission. Thus, similarities between parasites in their abundance, transmission biology, and phenology seem sufficient to explain species co-occurrences without invoking other processes such as species interactions. Received: 18 March 1996 / Accepted: 20 April 1999     

Occurrence of trichothecenes, zearalenone and ochratoxin A in cereals and mixed feed from central Lithuania

A total of 92 samples — 23 winter wheat, 12 summer barley, 5 oats and 52 mixed feed — were collected from a state factory in Kaunas, Lithuania and were analysed for the presence of trichothecenes, zearalenone (ZEN) and ochratoxin A (OA) using gas chromatography with electron capture detection and immunoaffinity column/high performance liquid chromatography with fluorescence and UV detections. Deoxynivalenol (DON), nivalenol (NIV), T-2 toxin and HT-2 toxin were detected at concentrations above 10 μg/kg in 68%, 48%, 38% and 8% of cereal samples, respectively, and in 98%, 88%, 12% and 8% of samples of mixed feed for swine and poultry. More than 10 μg/kg of zearalenone and ochratoxin A were found in 58% and 92% of the mixed feed samples, respectively. The highest concentrations of all analysed trichothecenes in Lithuanian mixed feed and cereal grains, with an exception of T-2 toxin in one oat lot and one sample of mixed feed and OA in two mixed feed samples, were lower than those reported as Lithuanian advisory or tolerance limits.     

Penicillium verrucosum in wheat and barley indicates presence of ochratoxin A

AIMS: The aims of this study were to isolate and identify ochratoxin A (OTA) producing fungi in cereals containing OTA and to determine the best selective and indicative medium for recovery of OTA producing fungi. METHODS AND RESULTS: Seventy-six wheat, barley and rye samples from Europe containing OTA and 17 samples without OTA were investigated using three different media, dichloran yeast sucrose agar (DYSG), dichloran rose bengal yeast extract sucrose agar (DRYES) and dichloran 18% glycerol agar (DG18). Hundred kernels were plated on each medium and the kind and number of fungal OTA producers were recorded as percentage of infestation. Penicillium verrucosum was the sole OTA producer found in cereals. The average percentage of infestation of P. verrucosum counts was recorded as 28.3% on DYSG, 10.3% on DRYES and 9.9% on DG18 on the OTA containing samples and 0.8% on DYSG, 0.4% on DRYES and 0.6% on DG18 for the samples without OTA. CONCLUSIONS: Penicillium verrucosum was the sole OTA producer in European cereals. Determination of P. verrucosum infestation and infection was best detected on DYSG after 7 days at 20 degrees C. The percentage of infestation of P. verrucosum found on DYSG and OTA content in cereals were correlated. More than 7% infestation of P. verrucosum indicated OTA contamination. SIGNIFICANCE AND IMPACT OF THE STUDY: The developed method could be used as a cereal quality control.     

Transformation of the mycotoxin ochratoxin A in plants. 2. Time course and rates of degradation and metabolite production in cell-suspension cultures of different crop plants

Ochratoxin A, one of the most toxic mycotoxins, can be metabolized nearly completely by suspension cultures of various plant cells. The transformation products identified in this study were almost the same in the cell-suspension cultures of maize, carrot, tomato, potato, soybean, wheat and barley, but the quantitative distribution differed strongly depending on incubation time and species of plant-cell culture. The compounds were extracted with ethyl acetate and detected by reversed-phase HPLC with gradient elution. From the result it is supposed that besides ochratoxin A also ochratoxin derivatives may occur in food and feedstuff of plant origin.     

Decline of Canada geese (Branta canadensis) and common goldeneye (Bucephala clangula) associated with a collapse of eelgrass (Zostera marina) in a Nova Scotia estuary

Mean numbers of migrant Canada geese (Branta canadensis) in Antigonish Harbour in the southern Gulf of St. Lawrence (Canada) during October to December were similar (approx. 450–500 birds) for the period 1998–2000. Similarly, during this period, geese used two foraging sites. However, in 2001, the average number of birds decreased by half and the primary foraging sites were used only rarely. This coincided with a decline of about 95% in the biomass of roots and rhizomes of eelgrass (Zostera marina) that occurred between October 2000 and 2001. Eelgrass is the principal food of geese in this estuary. In addition, there was a reduction of around 50% in the numbers of common goldeneye (Bucephala clangula), which feed on invertebrates associated with eelgrass. Lower than usual weekly abundances of geese and goldeneye are probably the result of an unusually short residence time in the estuary, rather than a decline in the total number of visiting migrants. We attribute these changes in the distribution and abundance of geese and goldeneyes to the dramatic decline in eelgrass. Electronic Publication     

半胱胺对成年鹅血浆中生长抑素和某些代谢激素的影响

目的:研究半胱胺(CS)对成年鹅生长抑素(SS)和某些代谢激素的影响.方法:14只装有翅静脉瘘管的成年杂交鹅(川白×太湖),经瘘管采取对照期和处理(一次性添喂100 mg/kg bw的半胱胺)后第1、3、5、7 d的血样,用RIA双抗法测定其中激素的含量.结果:实验期第1、3、5和7 d的生长抑素的含量均显著低于对照期(1.89±0.10 μg/L).促甲状腺激素较对照期(2.45±0.31 mIU/L)在第1 d显著下降21.63 %(P<0.05),第3、5 d均降低18.37%(P>0.05),第7 d基本回复正常(P>0.05). 实验期1、3、5和7 d的T4 较对照期(5.41±0.98 μg/L)显著升高(P<0.01);实验期第3 d的T3水平较对照期(1.05±0.06 μg /L)高36.19 %(P<0.01);同样,胰岛素在实验期的第5 d显著高于对照期(P<0.05).结论:CS能够降低成年鹅血液中SS含量,使T4、T3和胰岛素水平升高,提高了机体的代谢水平,同化作用加强,从而有利于生长.     

Combinatory effects of citrinin and ochratoxin A in immortalized human proximal tubule cells

Ochratoxin A (OTA) and citrinin (CIT) are two mycotoxins often occurring together in grains and cereals. Although both are nephrotoxic and can induce apoptosis, combination effects have not been examined up to now. Therefore, the aim of this study was to take a close look at the interactions of citrinin and OTA in cultured human proximal tubule-derived cells (IHKE cells). The cytotoxicity of both mycotoxins was studied, measuring the metabolic activity and the cell number. Furthermore, caspase 3-activation as a marker for apoptosis was examined for both mycotoxin alone and in combination. The results show that citrinin had an antagonistic effect on ochratoxin A induced caspase 3-activation in concentrations of 2.5 and 5 μmol/l. Higher concentrations (7.5 and 15 μmol/l) lead to additive effects, lower citrinin concentrations (0.25 and 1 μmol/l) did not show any effect at all. The observed decrease in caspase 3-activity was specific for the combination with OTA, since the combination of citrinin with cisplatin did not show any effect. Citrinin did not influence of the OTA-induced apoptosis when added two hours after applying ochratoxin A. Also the combination of both toxins decreased the uptake of OTA into the cells which might be an explanation for the antagonistic effect of citrinin in certain concentrations. However, the transport into cells can not be the only explanation. so further examinations are necessary. Presented at the 27^th Mykotoxin-Workshop. Dortmund, Germany, June 13–15, 2005.     

Heart rate modulation in bystanding geese watching social and non-social events

Simply observing other individuals interacting has been shown to affect subsequent behaviour and also hormones in 'bystander' individuals. However, immediate physiological responses of an observer have been hardly investigated. Here we present results on individuals' heart rate (HR) responses during various situations, which occur regularly in a flock of greylag geese (Anser anser, e.g. agonistic encounters, vehicles passing by). We recorded simultaneously HR and behaviour of 21 semi-tame free-roaming geese, equipped with fully implanted transmitters. We considered 304 social and 81 non-social events during which the focal individuals did not respond behaviourally. Independent of the spatial distance to the event, these HR responses were significantly greater in social contexts (e.g. departing or landing geese, agonistic interactions) than in non-social situations (e.g. vehicles passing by, thunder). Focal individuals showed a significantly higher maximum HR as well as a greater HR increase in response to agonistic interactions, in which the pair partner or a family member was involved, as compared with a non-affiliated goose. Also, HR was significantly higher when the bystander watched non-affiliated geese interacting, which were higher ranking than the focal. We conclude that these differences are due to different relevance of the recorded events for the focal individual, depending on the individuals involved in the observed interaction.     

Brief in vitro study on Botrytis cinerea and Aspergillus carbonarius regarding growth and ochratoxin A

Aims: To evaluate the effect of Botrytis cinerea growth on ochratoxin A (OTA) production by Aspergillus carbonarius and degradation. Methods and Results: OTA‐producing A. carbonarius and B. cinerea were grown on grape‐like medium at 20°C for 7 days. Radii of colonies were daily recorded and OTA was analysed. In addition, each B. cinerea isolate was inoculated on grape‐like synthetic nutrient medium (SNM) paired with each A. carbonarius isolate at a distance of 45 mm. Botrytis cinerea isolates were also grown in OTA‐spiked SNM. Growth rates of B. cinerea and A. carbonarius were 20 and 7·5 mm day^?1, respectively. The growth of the colonies of each species stopped when they contacted each other in paired cultures. OTA production by A. carbonarius in the contact area was affected by B. cinerea, but no clear trend was observed. All B. cinerea isolates showed to degrade between 24·2% and 26·7% of OTA from spiked SNM. Conclusions: The ecological advantage of B. cinerea, in terms of growth rate, vs. OTA‐producing Aspergillus in some wine‐growing regions and its ability to degrade OTA may explain the low levels of this toxin in noble wines. Significance and Impact of the Study: At determinate conditions, the presence of B. cinerea in grapes with A. carbonarius may help in reducing OTA accumulation.     

Repair of ochratoxin A-induced DNA damage and modulation of OTA-related genotoxicity by co-incubation with bile acids and methotrexatein vitro

The mycotoxin ochratoxin A (OTA) induced DNA strand breaks in porcine urinary bladder epithelial cells (PUBEC) and in Madin Darby canine kidney (MDCK) cells. A co-incubation with bile acids or methotrexate reduced or even prevented this adverse effect of OTAin vitro. The protective effect is possibly attributable to a decreased OTA uptake in cells, since bile acids and methotrexate are known to share common transport systems such as organic anion transporters (OAT) and/or organic anion transporting polypeptides (OATP) with the mycotoxin. OTA uptake in cells and its modulation can be one factor which determines the extent of adverse effects in different cell types. Another aspect of interest in this regard relates to repair of DNA damage: PUBEC cells are sensitive to OTA-induced damage which is more pronounced when DNA repair is blocked (by cytosine β-D-arabino-furanosid/hydroxyurea). On the other hand, when cells are kept in fresh (toxin-free) medium for 3 h, OTA-induced DNA damage decreased to control levels. Presented at the 26^th Mykotoxin-Workshop in Herrsching, Germany, May 17–19, 2004     

Estimation of genetic parameters for growth,carcass and overfeeding traits in a white geese strain

In an experimental strain of white plumage geese created in 1989, two experiments were carried out from 1993 to 1995 in order to estimate genetic parameters for growth, and carcass composition traits in non-overfed animals, and genetic parameters for growth and fatty liver formation in overfed animals. Four hundred and thirty-one non-overfed animals were bred and slaughtered at 11 weeks of age; they were measured for forearm length, keel bone length, chest circumference and breast depth before and after slaughtering. The carcasses were partly dissected in order weigh breast, breast muscle and skin + fat, and abdominal fat. Four hundred and seventy-seven overfed animals were slaughtered at 20 weeks of age; they were measured for "paletot" (breast meat, bone and meat from wings, bone and meat from thigh and legs) weight and liver weight. In these two experiments, the weights had moderate to high heritability values. Breast depth measured on live animals showed a low heritability value. In overfed animals, liver weight showed a high heritability value. Liver weight could be increased by selection without a great effect on "paletot" weight. Thus, obtaining a white plumage geese strain for fatty liver production by selection would be difficult because only 20% of overfed animals had fatty liver. The results did not allow to conclude on the influence of selection on liver weight on carcass traits such as muscle or fatty tissue weight.     

The dynamics of landscape change and snow geese in mid-continent North America

The Mid‐Continent Population of the lesser snow goose, which breeds in the eastern and central Canadian Arctic and sub‐Arctic, and winters in the southern United States and northern Mexico has increased 5–7% annually from the late 1960s to the mid‐1990s, largely because of increased survival in response to an agricultural food subsidy. The rise in numbers complements the increased use of nitrogen fertilizers and a corresponding rise in yields of rice, corn, and wheat along the flyways and on the wintering grounds. In sub‐Arctic migration areas and at Arctic breeding colonies, foraging by high numbers of birds has led to loss of coastal vegetation, adverse changes in soil properties and the establishment of an alternative stable state of exposed sediment, which can be detected with LANDSAT imagery. At a local scale, gosling growth, size and survival decreased in affected areas and other taxa have been adversely affected. The food subsidy on wintering and migration areas appears insufficient to meet reproductive demands as foraging in spring continues to occur on southern Hudson Bay staging and nesting areas. The recent introduction of liberal hunting regulations may reduce population size in the near term, but the revegetation of these coastal ecosystems will take decades to achieve. The present pattern of vegetation loss in these Arctic coastal systems is likely to continue in the forseeable future.