Determination of Zearalenone and Trichothecenes, Including Deoxynivalenol and Its Acetylated Derivatives, Nivalenol, T-2 and HT-2 Toxins, in Wheat and Wheat Products by LC-MS/MS: A Collaborative Study.

An analytical method for the simultaneous determination of trichothecenes-namely, nivalenol (NIV), deoxynivalenol (DON) and its acetylated derivatives (3- and 15-acetyl-DON), T-2 and HT-2 toxins-and zearalenone (ZEN) in wheat, wheat flour, and wheat crackers was validated through a collaborative study involving 15 participants from 10 countries. The validation study, performed within the M/520 standardization mandate of the European Commission, was carried out according to the IUPAC (International Union of Pure and Applied Chemistry) International Harmonized Protocol. The method was based on mycotoxin extraction from the homogenized sample material with a mixture of acetonitrile-water followed by purification and concentration on a solid phase extraction column. High-performance liquid chromatography coupled with tandem mass spectrometry was used for mycotoxin detection, using isotopically labelled mycotoxins as internal standards. The tested contamination ranges were from 27.7 to 378 µg/kg for NIV, from 234 to 2420 µg/kg for DON, from 18.5 to 137 µg/kg for 3-acetyl-DON, from 11.4 to 142 µg/kg for 15-acetyl-DON, from 2.1 to 37.6 µg/kg for T-2 toxin, from 6.6 to 134 µg/kg for HT-2 toxin, and from 31.6 to 230 µg/kg for ZEN. Recoveries were in the range 71-97% with the lowest values for NIV, the most polar mycotoxin. The relative standard deviation for repeatability (RSDr) was in the range of 2.2-34%, while the relative standard deviation for reproducibility (RSDR) was between 6.4% and 45%. The HorRat values ranged from 0.4 to 2.0. The results of the collaborative study showed that the candidate method is fit for the purpose of enforcing the legislative limits of the major Fusarium toxins in wheat and wheat-based products.

Subchronic (90-day) toxicity assessment of Somacy-FP100, a lipopolysaccharide-containing fermented wheat flour extract from Pantoea agglomerans.

Pantoea agglomerans is a Gram-negative bacterium that is ubiquitous in the environment, colonizing animals, humans, and numerous plants, including cotton and wheat. A lipopolysaccharide-containing fermented wheat flour extract from P. agglomerans (Somacy-FP100) is proposed for use as a food ingredient for individuals seeking foods for healthy aging. Previously published genotoxicity studies with Somacy-FP100 reported its lack of genotoxicity in vitro, but a subchronic toxicity study has not yet been performed. Therefore, to demonstrate the safety of Somacy-FP100 for use as a food ingredient, a 90-day oral (gavage) toxicity study in rats was conducted. Male and female Han Wistar rats were administered vehicle (control) or Somacy-FP100 at 500, 1500, or 4500 mg/kg body weight/day at a dose volume of 10 mL/kg body weight, for at least 90 days. No test article-related adverse clinical signs or effects on body weight, food consumption, or clinical pathology were observed, and there were no macroscopic or microscopic findings related to the test article. Therefore, 4500 mg/kg body weight/day (the highest dose tested and highest feasible dose) was established as the no-observed-adverse-effect level. This absence of subchronic toxicity, in addition to the previously reported lack of genotoxicity, demonstrates the safety of Somacy-FP100 for use as a food ingredient.

A critical review of the postulated role of the non-essential amino acid, ß-N-methylamino-L-alanine, in neurodegenerative disease in humans.

The compound BMAA (ß-N-methylamino-L-alanine) has been postulated to play a significant role in four serious neurological human diseases: Amyotrophic Lateral Sclerosis/Parkinsonism Dementia Complex (ALS/PDC) found on Guam, and ALS, Parkinsonism, and dementia that occur globally. ALS/PDC with symptoms of all three diseases first came to the attention of the scientific community during and after World War II. It was initially associated with cycad flour used for food because BMAA is a product of symbiotic cycad root-dwelling cyanobacteria. Human consumption of flying foxes that fed on cycad seeds was later suggested as a source of BMAA on Guam and a cause of ALS/PDC. Subsequently, the hypothesis was expanded to include a causative role for BMAA in other neurodegenerative diseases including Alzheimer's disease (AD) through exposures attributed to proximity to freshwaters and/or consumption of seafood due to its purported production by most species of cyanobacteria. The hypothesis that BMAA is the critical factor in the genesis of these neurodegenerative diseases received considerable attention in the medical, scientific, and public arenas. This review examines the history of ALS/PDC and the BMAA-human disease hypotheses; similarities and differences between ALS/PDC and the other diseases with similar symptomologies; the relationship of ALS/PDC to other similar diseases, studies of BMAA-mediated effects in lab animals, inconsistencies and data gaps in the hypothesis; and other compounds and agents that were suggested as the cause of ALS/PDC on Guam. The review concludes that the hypothesis of a causal BMAA neurodegenerative disease relationship is not supported by existing data.

Occurrence of mycotoxins in spelt and common wheat grain and their products.

Organic farming does not allow the use of conventional mineral fertilizers and crop protection products. As a result, in our experiments we chose to grow different species of cereals and to see how cereal species affect mycotoxin accumulation. This study describes the occurrence of deoxynivalenol (DON), zearalenone (ZEA) and T-2/HT-2 toxin in a survey of spelt and common wheat and their bran as well as flour. The analysis was conducted using an enzyme-linked immunosorbent assay (ELISA) method. The concentrations of DON, ZEA and T-2/HT-2 in Triticum spelta and T. aestivum were influenced by species, cereal type and year interaction. The highest concentrations of these mycotoxins were found in spelt grain with glumes, in spelt glumes and in spring wheat. These results show significantly higher concentrations of Fusarium toxins in glumes than in dehulled grain, which indicates the possible protective effect of spelt wheat glumes. The lowest DON, ZEA and T-2/HT-2 concentrations were determined in spelt grain without glumes. The research shows that it is potentially risky to produce bran from grain in which mycotoxin concentrations are below limits by European Union Regulation No. 1881/2006, since the concentration of mycotoxins in bran can be several times higher than that in grain. As a result, although bran is a dietary product characterised by good digestive properties, it can become a harmful product that can cause unpredictable health damage.

[Occupational asthma related to wheat. Presentation of one case]. / Asma laboral por trigo. Presentación de un caso.

Allergy to wheat proteins is the most common type of occupational asthma caused by cereals. Its incidence based on international reports is about 1-2.4/1000 cases among workers per year. In Mexico there is an underdiagnosis of occupational asthma > 99%. During the period 2009-2012 we recorded 5 cases of occupational asthma, 40% of them caused by cereals. This is the case of a 38 year old male, baker for 24 years exposed to wheat flour. In his workplace he used to wear surgical mask and there are no exhaust fans. He initiated 10 years ago with sneezing, itching, hyaline rhinorrhea and nasal obstruction. The following year he presented mild to moderate dyspnea, chest tightness, cough and wheezing during working hours, which improved during his days off. Pulmonary function tests were confirmatory of asthma, with negative skin prick tests to aeroallergens. Specific bronchial challenge was performed with wheat flour resulting positive with a decrease in VEF1 greater than 20%. Pharmacological treatment was provided and was sent to occupational medicine service for evaluation. The case was qualified as an occupational disease with a permanent partial disability of 25%, receiving a financial compensation. He returned to work in the same company but in a different area without wheat flour exposure. The diagnosis of occupational asthma is mainly clinical, according to Brooks¥ criteria, and a bronchial positive challenge test will confirms the diagnosis.

Metals in wheat flour; comparative study and safety control.

Cereal industry and its derived products have a big economic and social importance worldwide. Therefore, as wheat flour is a commodity for all bread and bakery industry, it is safety is of high nutrition and toxicological interest. In this investigation we intend to study and determine the content of twelve metals in 50 samples of wheat flour coming from a wheat flour industry. Macro elements sodium, potassium magnesium and calcium, micro elements manganese, iron, copper, zinc, chrome and nickel as well as toxic trace elements cadmium and lead have been analysed. The estimated diary intake of each metal and their contribution in percentage terms to the RDI (macro and micro elements) and to the PTWI (toxic elements) has been determined. Contribution of Cd and Pb to the PTWI was very low, a fact that shows safety in this wheat flour concerning toxic metals.

La industria de los cereales y sus derivados tiene una gran importancia económica y social en todo el mundo. Por ello, la seguridad de las harinas como materias primas de todas las industrias de panadería, bollería y repostería es de sumo interés nutricional y toxicológico. En este trabajo nos hemos propuesto estudiar y determinar el contenido de doce metales en 50 muestras de harina de trigo procedentes de una industria harinera. Se analizaron los macroelementos sodio, potasio, magnesio y calcio, los microelementos manganeso, hierro, cobre, zinc, cromo y níquel y los elementos traza tóxicos cadmio y plomo. Se determinó la ingesta diaria estimada de cada metal y su contribución porcentual a las IDRs (macro y microelementos) y a las PTWIs (elementos tóxicos). La contribución de las PTWIs para el Cd y Pb fue muy baja, lo que demuestra la seguridad de estas harinas en relación con estos metales tóxicos.

Assessing the impact of national level interventions on workplace respiratory disease in the UK: part 2--regulatory activity by the Health and Safety Executive.

OBJECTIVE: To investigate whether interventions implemented by the UK Health and Safety Executive addressing exposure to isocyanate-based spray paints in motor vehicle repair (MVR), flour dust in craft bakeries, rosin-based solder flux fume (RBSFF) in the electronics industry, metalworking fluids and wood dust coincided with a decline in incidence of work-related short latency respiratory disease (SLRD) or asthma in the target groups. METHOD: Changes in the incidence of SLRD reported to a UK-based surveillance scheme were compared using a longitudinal, negative binomial regression model with ß distributed random effects. An interrupted time series design was used and comparisons according to inclusion or exclusion in the target group were made by including a statistical interactions expressed as a ratio of incidence rate ratios (RIRRs) in the model. RESULTS: The incidence of SLRD attributed to flour dust significantly increased relative to all other agents (RIRR: 1.10; 95% CI 1.06 to 1.16) whereas SLRD attributed to RBSFF significantly declined relative to all other agents (0.94; 0.90 to 0.99). No significant changes in the incidence of SLRD attributed to wood dust (1.03; 0.91 to 1.16) or spray paints (1.03; 0.95 to 1.11) relative to all other agents were observed. A higher proportion of reports originated from the industries targeted by the intervention for RBSFF (65/107; 61%) than spray painting (27/93; 27%) or wood dust (16/42; 38%). CONCLUSIONS: These data support a beneficial effect of interventions to reduce exposure to RBSFF but an increase in SLRD attributed to flour dust may indicate increased exposure or increased awareness of the problem.

Monocrotophos poisoning through contaminated millet flour.

Several episodes of mass poisoning by organophosphates (OPs) have been reported from the developing countries. The diagnosis of OP-poisoning is mainly based on the characteristic clinical features and history of exposure to a known OP compound. Estimation of serum and red blood cell (RBC) cholinesterase activities are helpful in confirming the diagnosis. However, there is controversy regarding a definite relationship between serum cholinesterase activity and the severity of clinical manifestations and prognosis. This report describes an episode of mass monocrotophos poisoning that occurred due to accidental ingestion of monocrotophos-contaminated millet (so-called bavta) flour involving eight severely poisoned persons. Clinical presentation included severe abdominal pain, diarrhoea, vomiting, pupil narrowing, and difficulty breathing. On hospital admission, plasma cholinesterase (PChE) and especially RBC acetylcholinesterase (AChE) activities correlated well with clinical symptoms presented by the patients. This case study highlights the need for clinicians to be aware of OP-pesticide poisoning from food sources and the need to look for depressed PChE and AChE activities that may point to OP exposure, so that OP-poisoning can be identified immediately and patients can receive specific treatment, rather than general treatment for food poisoning.

Occupational rhinitis caused by concurrent sensitization to two different allergens.

BACKGROUND: Exposure to wheat flour and guar gum is a well-known cause of occupational respiratory allergies among workers in the food processing industry. To date, there have been no reports of occupational rhinitis (OR) caused concurrently by two different allergens present in the workplace. AIMS: To report a case of OR likely to be induced concurrently by exposure to wheat flour and guar gum in a mid-40s male employed in the food processing industry. METHODS: Allergy tests and nasal challenge tests were performed to investigate and confirm the diagnosis of OR. We discuss potential mechanisms involved in the observed dual sensitization. RESULTS: The patient showed positive responses to wheat and guar gum extracts on skin prick testing. The total IgE was 1680 kU/l (0-100 kU/l). The diagnosis of OR was confirmed by nasal challenge tests with wheat flour and guar gum on different days. In contrast to the control day, the challenge with flour and guar gum induced an immediate clinical reaction associated with a decrease in nasal volume measured by acoustic rhinometry. The patient was advised to avoid exposure to wheat and guar gum, which resulted in a gradual resolution of nasal symptoms. CONCLUSIONS: Co-sensitization and cross-reactivity are possible mechanisms involved in cases of concurrent sensitization to related and unrelated allergens in patients complaining of work-related rhinitis symptoms.

Exposure to flour dust and sensitization among bakery employees.

BACKGROUND: The National Institute for Occupational Safety and Health conducted a study to determine prevalences of sensitization to bakery-associated antigens (BAAs) and work-related respiratory symptoms at a large commercial bakery. METHODS: The following measurements were carried out: personal breathing zone (PBZ) and general area (GA) monitoring for inhalable flour dust, α-amylase and wheat, a questionnaire, and blood tests for IgE specific to flour dust, wheat, α-amylase, and common aeroallergens. RESULTS: Of 186 bakery employees present during our site visit, 161 completed the questionnaire and 96 allowed their blood to be drawn. The geometric mean PBZ and GA inhalable flour dust concentrations for the lower-exposure group was 0.235 mg/m(3), and for the higher-exposure group was 3.01 mg/m(3). Employees in the higher-exposure group had significantly higher prevalences of work-related wheezing, runny nose, stuffy nose, and frequent sneezing than the lower-exposure group. The prevalence of IgE specific to wheat was significantly higher among employees who ever had a job in the higher-exposure group or in production at another bakery at both the ≥ 0.10 kU/L and the ≥ 0.35 kU/L cutoffs, and to flour dust and α-amylase at the ≥ 0.10 kU/L cutoff, compared to the lower-exposure group. CONCLUSIONS: Despite knowledge of the risks of exposure to flour being available for centuries, U.S. employees are still at risk of sensitization and respiratory symptoms from exposure to high levels of BAA.

Environmental neurotoxin-induced progressive model of parkinsonism in rats.

OBJECTIVE: Exposure to a number of drugs, chemicals, or environmental factors can cause parkinsonism. Epidemiologic evidence supports a causal link between the consumption of flour made from the washed seeds of the plant Cycas micronesica by the Chamorro population of Guam and the development of amyotrophic lateral sclerosis/parkinsonism dementia complex. METHODS: We now report that consumption of washed cycad flour pellets by Sprague-Dawley male rats induces progressive parkinsonism. RESULTS: Cycad-fed rats displayed motor abnormalities after 2 to 3 months of feeding such as spontaneous unilateral rotation, shuffling gait, and stereotypy. Histological and biochemical examination of brains from cycad-fed rats revealed an initial decrease in the levels of dopamine and its metabolites in the striatum (STR), followed by neurodegeneration of dopaminergic (DAergic) cell bodies in the substantia nigra (SN) pars compacta (SNc). alpha-Synuclein (alpha-syn; proteinase K-resistant) and ubiquitin aggregates were found in the DAergic neurons of the SNc and neurites in the STR. In addition, we identified alpha-syn aggregates in neurons of the locus coeruleus and cingulate cortex. No loss of motor neurons in the spinal cord was found after chronic consumption of cycad flour. In an organotypic slice culture of the rat SN and the striatum, an organic extract of cycad causes a selective loss of dopamine neurons and alpha-syn aggregates in the SN. INTERPRETATION: Cycad-fed rats exhibit progressive behavioral, biochemical, and histological hallmarks of parkinsonism, coupled with a lack of fatality.

Previous studies underestimate BMAA concentrations in cycad flour.

The traditional diet of the Chamorro people of Guam has high concentrations of the neurotoxin BMAA, beta-methyl-amino-L-alanine, in cycad tortillas and from animals that feed on cycad seeds. We measured BMAA concentration in washed cycad flour and compared different extraction methods used by previous researchers in order to determine how much BMAA may have been unaccounted for in prior research. Samples were analyzed with AQC precolumn derivatization using HPLC-FD detection and verified with UPLC-UV, UPLC-MS, and triple quadrupole LC/MS/MS. Although previous workers had studied only the free amino acid component of BMAA in washed cycad flour, we detected significant levels of protein-associated BMAA in washed cycad flour. These data support a link between ALS/PDC and exposure to BMAA.

A dynamic population-based model for the development of work-related respiratory health effects among bakery workers.

OBJECTIVES: This paper presents a dynamic population-based model for the development of sensitisation and respiratory symptoms in bakery workers. The model simulates a population of individual workers longitudinally and tracks the development of work-related sensitisation and respiratory symptoms in each worker. METHODS: The model has three components: a multi-stage disease model describing the development of sensitisation and respiratory symptoms in each worker over time; an exposure model describing occupational exposure to flour dust and allergens; and a basic population model describing the length of a worker's career in the bakery sector and the influx of new workers. Each worker's disease state is modelled independently using a discrete time Markov Chain, updated yearly using each individual's simulated exposure. A Bayesian analysis of data from a recent epidemiological study provided estimates of the yearly transition probabilities between disease states. RESULTS: For non-atopic/non-sensitised workers the estimated probabilities of developing moderate (upper respiratory) symptoms and progression to severe (lower respiratory) symptoms are 0.4% (95% CI 0.3 to 0.5%) and 1.1% (95% CI 0.6 to 1.9%) per mg/m(3)/year of flour dust, respectively, and approximately twice these for atopic workers. The model predicts that 36% (95% CI 26 to 46%) of workers with severe symptoms are sensitised to wheat and 22% (95% CI 12 to 37%) to alpha-amylase. The predicted mean latency period for respiratory symptoms was 10.3 years (95% CI 8.3 to 12.3). CONCLUSIONS: While the model provides a valuable population-level representation of the mechanisms contributing to respiratory diseases in bakers, it was primarily developed for use in quantitative health impact assessment. Future research will use the model to evaluate a range of workplace interventions, including achievable reductions in exposure and health surveillance. The general methodology is applicable to other diseases such as chronic obstructive pulmonary disease, silicosis and musculoskeletal disorders and could be particularly valuable for forecasting changes in long latency diseases.

Rye flour allergens: an emerging role in baker's asthma.

BACKGROUND: Exposure to wheat flour is usually considered the most important cause of baker's asthma. However, other flours frequently used in bakeries may play an emerging role as relevant allergens causing occupational asthma. AIMS OF STUDY: We report on two cases of baker's asthma mainly caused by exposure to rye flour. The profile of allergen sensitization to cereal flour was investigated. METHODS: Two bakery workers suffering from rhinoconjunctivitis and asthma symptoms at work underwent an in vivo study (skin prick tests and bronchial allergen challenge) and in vitro study (total serum IgE, specific serum IgE and immunoblotting). RESULTS: Specific inhalation challenge with wheat flour did not elicit an asthmatic reaction, however both patients showed an early asthmatic reaction with the rye flour challenge. Rye flour-immunoblotting showed IgE-binding bands around 12-15 kDa, that correspond to rye flour enzymatic inhibitors which were not present in the wheat flour immunoblot. CONCLUSIONS: Both bakers had developed occupational asthma to rye flour (confirmed by specific inhalation challenge test). Rye flour allergens (enzymatic inhibitors) are important allergens that should be considered in the diagnosis of baker's asthma.

Occupational eosinophilic bronchitis in a foundry worker exposed to isocyanate and a baker exposed to flour.

Eosinophilic bronchitis without asthma may occur as a consequence of occupational exposure. The cases of a foundry worker and a baker who developed symptoms, respectively, due to exposure to isocyanate and flour, are reported. Cough was not associated with variable airflow obstruction or with airway hyper-responsiveness and was responsive to inhaled corticosteroids. The eosinophilia detectable in their sputum was causally related to the occupational exposure in the workplace. The examination of induced sputum should be used in addition to the objective monitoring of lung function for workers who have asthma-like symptoms in an occupational setting.