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Toxins, Volume 11, Issue 8 (August 2019)

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Cover Story (view full-size image) A massive Ostreopsis cf. ovata bloom was described in Currais Archipelago, a poorly studied region [...] Read more.
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Open AccessArticle
Biotransformation of the Mycotoxin Zearalenone to its Metabolites Hydrolyzed Zearalenone (HZEN) and Decarboxylated Hydrolyzed Zearalenone (DHZEN) Diminishes its Estrogenicity In Vitro and In Vivo
Received: 26 July 2019 / Revised: 9 August 2019 / Accepted: 16 August 2019 / Published: 20 August 2019
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Abstract
Zearalenone (ZEN)-degrading enzymes are a promising strategy to counteract the negative effects of this mycotoxin in livestock. The reaction products of such enzymes need to be thoroughly characterized before technological application as a feed additive can be envisaged. Here, we evaluated the estrogenic [...] Read more.
Zearalenone (ZEN)-degrading enzymes are a promising strategy to counteract the negative effects of this mycotoxin in livestock. The reaction products of such enzymes need to be thoroughly characterized before technological application as a feed additive can be envisaged. Here, we evaluated the estrogenic activity of the metabolites hydrolyzed zearalenone (HZEN) and decarboxylated hydrolyzed zearalenone (DHZEN) formed by hydrolysis of ZEN by the zearalenone-lactonase Zhd101p. ZEN, HZEN, and DHZEN were tested in two in vitro models, the MCF-7 cell proliferation assay (0.01–500 nM) and an estrogen-sensitive yeast bioassay (1–10,000 nM). In addition, we compared the impact of dietary ZEN (4.58 mg/kg) and equimolar dietary concentrations of HZEN and DHZEN on reproductive tract morphology as well as uterine mRNA and microRNA expression in female piglets (n = 6, four weeks exposure). While ZEN increased cell proliferation and reporter gene transcription, neither HZEN nor DHZEN elicited an estrogenic response, suggesting that these metabolites are at least 50–10,000 times less estrogenic than ZEN in vitro. In piglets, HZEN and DHZEN did not increase vulva size or uterus weight. Moreover, RNA transcripts altered upon ZEN treatment (EBAG9, miR-135a-5p, miR-187-3p and miR-204-5p) were unaffected by HZEN and DHZEN. Our study shows that both metabolites exhibit markedly reduced estrogenicity in vitro and in vivo, and thus provides an important basis for further evaluation of ZEN-degrading enzymes. Full article
(This article belongs to the Special Issue Novel Approaches to Minimising Mycotoxin Contamination)
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Open AccessArticle
Immunohistochemical Review of Leydig Cell Lesions in Ochratoxin A-Treated Fischer Rats and Controls
Received: 4 July 2019 / Revised: 22 July 2019 / Accepted: 12 August 2019 / Published: 20 August 2019
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Abstract
Ochratoxin A is best known as a potent renal carcinogen in male rats and mice after necessarily protracted ingestion, although valid extrapolation to any human disease has not been verified. The hypothesis that the toxin is a cause of human testicular cancer was [...] Read more.
Ochratoxin A is best known as a potent renal carcinogen in male rats and mice after necessarily protracted ingestion, although valid extrapolation to any human disease has not been verified. The hypothesis that the toxin is a cause of human testicular cancer was proposed a decade ago and has proliferated since, partly through incomplete study of the scientific literature. Archived tumorous rat testes were available from Fischer F344 rats exposed to continuous dietary exposure for half of or the whole life in London in the 2000s. Renal cancer occurred in some of these cases and testicular tumours were observed frequently, as expected, in both treated and untreated animals. Application of clinical immunohistochemistry has for the first time consistently diagnosed the testicular hypertrophy in toxin-treated rats as Leydig cell tumours. Comparison is made with similar analysis of tumorous testes from control (untreated) rats from U.S. National Toxicology Program studies, both of ochratoxin A (1989) and the more recent one on Ginkgo biloba. All have been found to have identical pathology as being of sex cord-stromal origin. Such are rare in humans, most being of germinal cell origin. The absence of experimental evidence of any specific rat testicular cellular pathology attributable to long-term dietary ochratoxin A exposure discredits any experimental animal evidence of testicular tumorigenicity. Thus, no epidemiological connection between ochratoxin A and the incidence of human testicular cancer can be justified scientifically. Full article
(This article belongs to the Special Issue Ochratoxin in Food Safety and Public Health)
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Open AccessArticle
Perennial Ryegrass Alkaloids Increase Respiration Rate and Decrease Plasma Prolactin in Merino Sheep under Both Thermoneutral and Mild Heat Conditions
Received: 19 July 2019 / Revised: 14 August 2019 / Accepted: 16 August 2019 / Published: 19 August 2019
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Abstract
A study was undertaken to determine the effects of feeding two levels of perennial ryegrass alkaloids (nil vs. moderate) under two climatic conditions. Alkaloids were fed via endophyte-infected perennial ryegrass seed and hay. Twenty-four Merino ewe weaners (six months, initial BW 32 ± [...] Read more.
A study was undertaken to determine the effects of feeding two levels of perennial ryegrass alkaloids (nil vs. moderate) under two climatic conditions. Alkaloids were fed via endophyte-infected perennial ryegrass seed and hay. Twenty-four Merino ewe weaners (six months, initial BW 32 ± 1.7 kg) were used in a study that lasted for 21 days after 14 days of adaptation. Sheep were fed either a control or alkaloid (Alk, 110 μg/kg LW ergovaline and 75 μg/kg LW lolitrem B) supplemented diet. Sheep were exposed to either constant thermoneutral (TN, 21–22 °C, 49% RH) or mildly heated (HS, 33 °C 1000–1500 h, 28% relative humidity) conditions. Dietary Alk and HS reduced dry matter intake (DMI) (p < 0.001, p = 0.02, respectively) with the combination of both reducing DMI by 42%. Reductions in DMI resulted in a lower daily gain in the Alk treatment (p < 0.001). Feed digestibility was reduced in the combined treatment (p = 0.03). Rectal temperature, respiration rate, and skin temperature increased in the Alk treatment. Plasma prolactin concentrations were decreased by Alk and increased by mild HS. The data indicate that production is compromised in the presence of Alk and mild HS, with this effect being exacerbated by a combination of both. Full article
(This article belongs to the Special Issue Dietary Mycotoxin Exposure to Livestock and Poultry)
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Open AccessArticle
The Degradation of Deoxynivalenol by Using Electrochemical Oxidation with Graphite Electrodes and the Toxicity Assessment of Degradation Products
Received: 2 July 2019 / Revised: 30 July 2019 / Accepted: 9 August 2019 / Published: 19 August 2019
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Abstract
Deoxynivalenol (DON) is a common mycotoxin, which is known to be extremely harmful to human and livestock health. In this study, DON was degraded by electrochemical oxidation (ECO) using a graphite electrode and NaCl as the supporting electrolyte. The graphite electrode is advantageous [...] Read more.
Deoxynivalenol (DON) is a common mycotoxin, which is known to be extremely harmful to human and livestock health. In this study, DON was degraded by electrochemical oxidation (ECO) using a graphite electrode and NaCl as the supporting electrolyte. The graphite electrode is advantageous due to its electrocatalytic activity, reusability, and security. The degradation process can be expressed by first-order kinetics. Approximately 86.4% of DON can be degraded within 30 min at a potential of 0.5 V. The degradation rate reached 93.2% within 30 min, when 0.5 V potential was used for electrocatalyzing a 10 mg/L DON solution. The degradation rate of DON in contaminated wet distiller’s grain with solubles (WDGS) was 86.37% in 60 min. Moreover, results from the cell counting kit-8 (CCK-8) and 4,6-diamidino-2-phenylindole dihydrochloride (DAPI) staining assay indicated that ECO reduced the DON-induced cytotoxicity and apoptotic bodies in a gastric epithelial cell line (GES-1) compared to the DON-treated group. These findings provide new insights into the application of ECO techniques for degrading mycotoxins, preventing food contamination, and assessing DON-related hazards. Full article
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Open AccessArticle
Suppressive Effects of Bee Venom-Derived Phospholipase A2 on Mechanical Allodynia in a Rat Model of Neuropathic Pain
Received: 10 June 2019 / Revised: 14 August 2019 / Accepted: 15 August 2019 / Published: 19 August 2019
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Abstract
Bee venom (BV) has a long history of being used in traditional Korean medicine to relieve pain. Here, we investigated the effect of BV-derived phospholipase A2 (bvPLA2), a major component of BV, on peripheral nerve injury-induced neuropathic pain in rats. Spinal nerve ligation [...] Read more.
Bee venom (BV) has a long history of being used in traditional Korean medicine to relieve pain. Here, we investigated the effect of BV-derived phospholipase A2 (bvPLA2), a major component of BV, on peripheral nerve injury-induced neuropathic pain in rats. Spinal nerve ligation (SNL) was performed in Sprague Dawley rats to induce neuropathic pain, and paw withdrawal thresholds were measured using von Frey test. Mechanical allodynia, the representative symptom of neuropathic pain, was manifested following SNL and persisted for several weeks. The repetitive bvPLA2 treatment (0.2 mg/kg/day, i.p.) for two days significantly relieved the SNL-induced mechanical allodynia. The antiallodynic effect of bvPLA2 was blocked by spinal pretreatment with α1-adrenergic antagonist prazosin (30 μg, i.t.) but not with α2-adrenergic antagonist idazoxan (50 μg, i.t.). Also, the spinal application of α1-adrenergic agonist phenylephrine (50 μg, i.t.) reduced mechanical allodynia. These results indicate that bvPLA2 could relieve nerve injury-induced neuropathic mechanical allodynia through the activation of spinal α1-adrenergic receptors. Full article
(This article belongs to the Special Issue From Toxins to Drugs)
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Open AccessArticle
Impact of Naturally Contaminated Substrates on Alphitobius diaperinus and Hermetia illucens: Uptake and Excretion of Mycotoxins
Received: 23 July 2019 / Revised: 6 August 2019 / Accepted: 16 August 2019 / Published: 18 August 2019
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Abstract
Insects are considered a suitable alternative feed for livestock production and their use is nowadays regulated in the European Union by the European Commission Regulation No. 893/2017. Insects have the ability to grow on a different spectrum of substrates, which could be naturally [...] Read more.
Insects are considered a suitable alternative feed for livestock production and their use is nowadays regulated in the European Union by the European Commission Regulation No. 893/2017. Insects have the ability to grow on a different spectrum of substrates, which could be naturally contaminated by mycotoxins. In the present work, the mycotoxin uptake and/or excretion in two different insect species, Alphitobius diaperinus (Lesser Mealworm, LM) and Hermetia illucens (Black Soldier Fly, BSF), grown on naturally contaminated substrates, was evaluated. Among all the substrates of growth tested, the Fusarium toxins deoxynivalenol (DON), fumonisin 1 and 2 (FB1 and FB2) and zearalenone (ZEN) were found in those based on wheat and/or corn. No mycotoxins were detected in BSF larvae, while quantifiable amount of DON and FB1 were found in LM larvae, although in lower concentration than those detected in the growing substrates and in the residual fractions. Mass balance calculations indicated that BSF and LM metabolized mycotoxins in forms not yet known, accumulating them in their body or excreting in the faeces. Further studies are required in this direction due to the future employment of insects as feedstuff. Full article
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Open AccessArticle
Bacteriocin Occurrence and Activity in Escherichia coli Isolated from Bovines and Wastewater
Received: 24 July 2019 / Revised: 7 August 2019 / Accepted: 12 August 2019 / Published: 15 August 2019
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Abstract
The increasing prevalence of antimicrobial resistant (AMR) E. coli and related Enterobacteriaceae is a serious problem necessitating new mitigation strategies and antimicrobial agents. Bacteriocins, functionally diverse toxins produced by most microbes, have long been studied for their antimicrobial potential. Bacteriocins have once again [...] Read more.
The increasing prevalence of antimicrobial resistant (AMR) E. coli and related Enterobacteriaceae is a serious problem necessitating new mitigation strategies and antimicrobial agents. Bacteriocins, functionally diverse toxins produced by most microbes, have long been studied for their antimicrobial potential. Bacteriocins have once again received attention for their role as probiotic traits that could mitigate pathogen burden and AMR bacteria in livestock. Here, bacteriocins were identified by activity screening and whole-genome sequencing of bacteriocin-producers capable of inhibiting bovine and wastewater E. coli isolates enriched for resistance to cephalosporins. Producers were tested for activity against shiga toxin-producing E. coli (STEC), AMR E. coli, and related enteric pathogens. Multiple bacteriocins were found in 14 out of 90 E. coli isolates tested. Based on alignment within BACTIBASE, colicins M, B, R, Ia, Ib, S4, E1, E2, and microcins V, J25, and H47, encoded by identical, variant, or truncated genes were identified. Although some bacteriocin-producers exhibited activity against AMR and STEC E. coli in agar-based assays, most did not. Despite this idiosyncrasy, liquid co-cultures of all bacteriocinogenic isolates with luciferase-expressing generic (K12) or STEC E. coli (EDL933) resulted in inhibited growth or reduced viability. These abundant toxins may have real potential as next-generation control strategies in livestock production systems but separating the bacteriocin from its immunity gene may be necessary for such a strategy to be effective. Full article
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Open AccessArticle
Structure Elucidation and Toxicity Analysis of the Degradation Products of Deoxynivalenol by Gaseous Ozone
Received: 26 July 2019 / Revised: 6 August 2019 / Accepted: 12 August 2019 / Published: 15 August 2019
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Abstract
Fusarium Head Blight (FHB) or scab is a fungal disease of cereal grains. Wheat scab affects the yield and quality of wheat and produces mycotoxins such as deoxynivalenol (DON), which can seriously threaten human and animal health. In this study, gaseous ozone was [...] Read more.
Fusarium Head Blight (FHB) or scab is a fungal disease of cereal grains. Wheat scab affects the yield and quality of wheat and produces mycotoxins such as deoxynivalenol (DON), which can seriously threaten human and animal health. In this study, gaseous ozone was used to degrade DON in wheat scab and the degradation products of ozonolysis were analyzed by ultra-performance liquid chromatography quadrupole-orbitrap mass spectrometry (UHPLC Q-Orbitrap). Toxicology analyses of the degradation products were also studied using structure-activity relationships. Ozone (8 mg L−1 concentration) was applied to 2 μg mL−1 of DON in ultrapure water, resulted in 95.68% degradation within 15 s. Ten ozonized products of DON in ultrapure water were analyzed and six main products (C15H18O7, C15H18O9, C15H22O9, C15H20O10, C15H18O8, and C15H20O9) were analyzed at varying concentrations of ozone and DON. Structural formulae were assigned to fragmentation products generated by MS2 and Mass Frontier® software. According to structure-activity relationship studies, the toxicities of the ozonized products were significantly decreased due to de-epoxidation and the attack of ozone at the C9-10 double bond in DON. Based on the results of the study above, we can find that gaseous ozone is an efficient and safe technology to degrade DON, and these results may provide a theoretical basis for the practical research of detoxifying DON in scabby wheat and other grains. Full article
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Open AccessArticle
The Effect of Low and High Dose Deoxynivalenol on Intestinal Morphology, Distribution, and Expression of Inflammatory Cytokines of Weaning Rabbits
Received: 24 June 2019 / Revised: 4 August 2019 / Accepted: 6 August 2019 / Published: 13 August 2019
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Abstract
Deoxynivalenol (DON) is a potential pathogenic factor to humans and animals, and intestinal tract is the primary target organ of DON. Data concerning the effects of DON on rabbits are scarce, especially for weaning rabbits. In this study, 45 weaning rabbits (35 d) [...] Read more.
Deoxynivalenol (DON) is a potential pathogenic factor to humans and animals, and intestinal tract is the primary target organ of DON. Data concerning the effects of DON on rabbits are scarce, especially for weaning rabbits. In this study, 45 weaning rabbits (35 d) were randomly and equally assigned into three groups. Group A was fed basic diet, while groups B and C were added DON at 0.5 mg/kg BW/d and 1.5 mg/kg BW/d, respectively, based on the basic diet. The experiment lasted for 24 days and the intestinal morphology, expression, and distribution of several cytokines in intestinal segments have been examined. The results indicated that ADG decreased while F/G increased significantly compared with the control group after DON added at 1.5 mg/kg BW/d. Some of the morphometric parameters (villi length, crypt depth, and goblet cells density) changed after DON was added. Meanwhile, the concentration as well as the expression levels of relative protein and mRNA of IL-1β, IL-2, IL-6, and IL-8 increased significantly. The immunohistochemistry results illustrated that the quantity and distribution of positive cells of inflammatory cytokines were changed after DON was added. In conclusion, the addition of DON damaged the intestinal morphology and changed the distribution and expression of inflammatory cytokines. The toxic effect depended on the dosage of DON. Full article
(This article belongs to the Special Issue Mycotoxins, Immunity, and Inflammation)
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Open AccessCommunication
Clostridium botulinum and Clostridium perfringens Occurrence in Kazakh Honey Samples
Received: 14 June 2019 / Revised: 8 August 2019 / Accepted: 9 August 2019 / Published: 13 August 2019
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Abstract
The aim of this study was to assess occurrence of Clostridium botulinum and Clostridium perfringens in honey samples from Kazakhstan. Analyses were carried out using a set of PCR methods for identification of anaerobic bacteria, and detection of toxin genes of C. botulinum [...] Read more.
The aim of this study was to assess occurrence of Clostridium botulinum and Clostridium perfringens in honey samples from Kazakhstan. Analyses were carried out using a set of PCR methods for identification of anaerobic bacteria, and detection of toxin genes of C. botulinum and C. perfringens. Among 197 samples, C. botulinum was noticed in only one (0.5%). The isolated strain of this pathogen showed the presence of the bont/A and ntnh genes. C. perfringens strains were isolated from 18 (9%) samples, and mPCR (multiplex PCR) analysis led to them all being classified as toxin type A with the ability to produce α toxin. Sequence analysis of 16S rDNA genes showed occurrence in 4 samples of other anaerobes related to C. botulinum, which were C. sporogenes and C. beijerinckii strains. C. botulinum prevalence in honey samples from Kazakhstan in comparison to the prevalence in samples collected from the other regions seems to be less. The highest prevalence of Clostridium sp. was noticed in the East Kazakhstan province. Our study is the first survey on BoNT-producing clostridia and C. perfringens prevalence in Kazakh honey. Full article
(This article belongs to the Special Issue New Challenges in Foodborne Botulism Outbreaks)
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Open AccessArticle
Determination of Ochratoxin A (OTA), Ochratoxin B (OTB), T-2, and HT-2 Toxins in Wheat Grains, Wheat Flour, and Bread in Lebanon by LC-MS/MS
Received: 7 July 2019 / Revised: 2 August 2019 / Accepted: 5 August 2019 / Published: 12 August 2019
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Abstract
Cereals are prone to fungal infection during growth, harvesting, transportation, and/or storage. As a result, cereals such as wheat grains and wheat-derived products may be contaminated with mycotoxins leading to acute and chronic health exposure. The current study investigated the presence of the [...] Read more.
Cereals are prone to fungal infection during growth, harvesting, transportation, and/or storage. As a result, cereals such as wheat grains and wheat-derived products may be contaminated with mycotoxins leading to acute and chronic health exposure. The current study investigated the presence of the mycotoxins: ochratoxin A (OTA), ochratoxin B (OTB), T-2, and HT-2 toxins in samples of wheat grains (n = 50), wheat flour (n = 50), and bread (n = 37) from the main mills in Lebanon using LC-MS/MS. Accuracy ranged from 98–100%, recoveries from 93–105%, and intraday and interday precision were 5–7% and 9–12%, respectively. The tested wheat grains, wheat flour, and bread samples did not contain detectable levels of T-2 and HT-2 toxins and OTB. Four wheat flour samples (8% of flour samples) showed positive OTA levels ranging from 0.6–3.4 μg·kg−1 with an arithmetic mean of 1.9 ± 0.2 μg·kg−1. Only one sample contained an OTA concentration greater than the limit set by the European Union (3 μg·kg−1) for wheat-derived products. This study suggests that mycotoxin contamination of wheat grains, wheat flour, and bread in Lebanon is currently not a serious public health concern. However, surveillance strategies and monitoring programs must be routinely implemented to ensure minimal mycotoxin contamination of wheat-based products. Full article
(This article belongs to the Section Mycotoxins)
Open AccessArticle
Contrasting Toxin Selectivity between the Marine Pufferfish Takifugu pardalis and the Freshwater Pufferfish Pao suvattii
Received: 10 July 2019 / Revised: 6 August 2019 / Accepted: 8 August 2019 / Published: 10 August 2019
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Abstract
To clarify the differences in toxin selectivity between marine and freshwater pufferfish, we conducted experiments in artificially reared nontoxic specimens of Takifugu pardalis (marine) and Pao suvattii (freshwater) using tetrodotoxin (TTX) and paralytic shellfish poison (PSP; decarbamoylsaxitoxin (dcSTX) or saxitoxin (STX)). T. pardalis [...] Read more.
To clarify the differences in toxin selectivity between marine and freshwater pufferfish, we conducted experiments in artificially reared nontoxic specimens of Takifugu pardalis (marine) and Pao suvattii (freshwater) using tetrodotoxin (TTX) and paralytic shellfish poison (PSP; decarbamoylsaxitoxin (dcSTX) or saxitoxin (STX)). T. pardalis specimens were administered feed homogenate containing TTX or dcSTX (dose of toxin, 55.2 nmol/fish) and P. suvattii specimens were administered feed homogenate containing TTX + STX (dose of each toxin, 19.2 nmol/fish) by oral gavage. The toxin content in the intestine, muscle, skin, liver, and gonads was quantified after 24 and 48 or 72 h. In T. pardalis, TTX administered into the intestine was absorbed into the body and transferred and retained mainly in the skin and liver, while dcSTX was hardly retained in the body, although it partly remained in the intestine. In strong contrast, in P. suvattii, little TTX remained in the body, whereas STX was absorbed into the body and was transferred and retained in the ovary and skin. The findings revealed that TTX/PSP selectivity differs between the marine species T. pardalis and the freshwater species P. suvattii. T. pardalis, which naturally harbors TTX, selectively accumulates TTX, and P. suvattii, which naturally harbors PSP, selectively accumulates PSP. Full article
(This article belongs to the Special Issue Isolation and Characterization of Marine Toxins)
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Open AccessArticle
Aflatoxin Exposure from Milk in Rural Kenya and the Contribution to the Risk of Liver Cancer
Received: 24 June 2019 / Revised: 13 July 2019 / Accepted: 25 July 2019 / Published: 10 August 2019
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Abstract
Milk is an important commodity in Kenya; the country has the largest dairy herd and highest per capita milk consumption in East Africa. As such, hazards in milk are of concern. Aflatoxin M1 (AFM1) is a toxic metabolite of aflatoxin [...] Read more.
Milk is an important commodity in Kenya; the country has the largest dairy herd and highest per capita milk consumption in East Africa. As such, hazards in milk are of concern. Aflatoxin M1 (AFM1) is a toxic metabolite of aflatoxin B1 (AFB1) excreted in milk by lactating animals after ingesting AFB1-contaminated feeds. This metabolite is injurious to human health, but there is little information on the risk to human health posed by AFM1 in milk in rural Kenya. To fill this gap, a quantitative risk assessment (QRA) applying probabilistic statistical tools to quantify risks was conducted. This assessed the risk of liver cancer posed by AFM1 in milk, assuming 10-fold lower carcinogenicity than AFB1. Data from four agro–ecological zones in Kenya (semi-arid, temperate, sub-humid and humid) were used. We estimated that people were exposed to between 0.3 and 1 ng AFM1 per kg body weight per day through the consumption of milk. The annual incidence rates of cancer attributed to the consumption of AFM1 in milk were 3.5 × 10−3 (95% CI: 3 × 10−3–3.9 × 10−3), 2.9 × 10−3 (95% CI: 2.5 × 10−3–3.3 × 10−3), 1.4 × 10−3 (95% CI: 1.2 × 10−3–1.5 × 10−3) and 2.7 × 10−3 (95% CI: 2.3 × 10−3–3 × 10−3) cancers per 100,000 in adult females, adult males, children 6–18 years old, and in children less than five years old, respectively. Our results show that aflatoxin exposure from milk contributes relatively little to the incidence of liver cancer. Nonetheless, risk managers should take action based on cumulative exposure from all sources of aflatoxins. Full article
(This article belongs to the Special Issue Dietary Mycotoxin Exposure: Emerging Risks to Human Health)
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Open AccessArticle
Paralytic Shellfish Toxin Uptake, Assimilation, Depuration, and Transformation in the Southeast Asian Green-Lipped Mussel (Perna viridis)
Received: 5 July 2019 / Revised: 2 August 2019 / Accepted: 6 August 2019 / Published: 9 August 2019
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Abstract
Bivalve molluscs represent an important food source within the Philippines, but the health of seafood consumers is compromised through the accumulation of harmful algal toxins in edible shellfish tissues. In order to assess the dynamics of toxin risk in shellfish, this study investigated [...] Read more.
Bivalve molluscs represent an important food source within the Philippines, but the health of seafood consumers is compromised through the accumulation of harmful algal toxins in edible shellfish tissues. In order to assess the dynamics of toxin risk in shellfish, this study investigated the uptake, depuration, assimilation, and analogue changes of paralytic shellfish toxins in Perna viridis. Tank experiments were conducted where mussels were fed with the toxic dinoflagellate Alexandrium minutum. Water and shellfish were sampled over a six day period to determine toxin concentrations in the shellfish meat and water, as well as algal cell densities. The maximum summed toxin concentration determined was 367 µg STX eq./100 g shellfish tissue, more than six times higher than the regulatory action limit in the Philippines. Several uptake and depuration cycles were observed during the study, with the first observed within the first 24 h coinciding with high algal cell densities. Toxin burdens were assessed within different parts of the shellfish tissue, with the highest levels quantified in the mantle during the first 18 h period but shifting towards the gut thereafter. A comparison of toxin profile data evidenced the conversion of GTX1,4 in the source algae to the less potent GTX2,3 in the shellfish tissue. Overall, the study illustrated the temporal variability in Perna viridis toxin concentrations during a modelled algal bloom event, and the accumulation of toxin from the water even after toxic algae were removed. Full article
(This article belongs to the Special Issue Marine Biotoxins and Seafood Poisoning)
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Open AccessArticle
Genetic Profiling of Aspergillus Isolates with Varying Aflatoxin Production Potential from Different Maize-Growing Regions of Kenya
Received: 22 June 2019 / Revised: 25 July 2019 / Accepted: 5 August 2019 / Published: 9 August 2019
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Abstract
Highly toxigenic strains of Aspergillus flavus have been reported to frequently contaminate maize, causing fatal aflatoxin poisoning in Kenya. To gain insights into the environmental and genetic factors that influence toxigenicity, fungi (n = 218) that were culturally identified as A. flavus [...] Read more.
Highly toxigenic strains of Aspergillus flavus have been reported to frequently contaminate maize, causing fatal aflatoxin poisoning in Kenya. To gain insights into the environmental and genetic factors that influence toxigenicity, fungi (n = 218) that were culturally identified as A. flavus were isolated from maize grains samples (n = 120) from three regions of Kenya. The fungi were further characterized to confirm their identities using a PCR-sequence analysis of the internal transcribed spacer (ITS) region of rDNA which also revealed all of them to be A. flavus. A subset of 72 isolates representing ITS sequence-based phylogeny cluster and the agroecological origin of maize samples was constituted for subsequent analysis. The analysis of partial calmodulin gene sequences showed that the subset consisted of A. flavus (87%) and Aspergillus minisclerotigenes (13%). No obvious association was detected between the presence of seven aflatoxin biosynthesis genes and fungal species or region. However, the presence of the aflD and aflS genes showed some association with aflatoxin production. The assessment of toxigenicity showed higher aflatoxin production potential in A. minisclerotigenes isolates. Given that A. minisclerotigenes were mainly observed in maize samples from Eastern Kenya, a known aflatoxin hotspot, we speculate that production of copious aflatoxin is an adaptative trait of this recently discovered species in the region. Full article
(This article belongs to the collection Aflatoxins)
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Open AccessFeature PaperArticle
Biomonitoring of Deoxynivalenol and Deoxynivalenol-3-glucoside in Human Volunteers: Renal Excretion Profiles
Received: 19 June 2019 / Revised: 2 August 2019 / Accepted: 6 August 2019 / Published: 8 August 2019
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Abstract
Biomarkers for the determination of the dietary exposure to deoxynivalenol (DON) have been proposed in the past but so far no quantification of their use in humans has been carried out. Following a human intervention study with two mycotoxins, namely DON and deoxynivalenol-3-glucoside [...] Read more.
Biomarkers for the determination of the dietary exposure to deoxynivalenol (DON) have been proposed in the past but so far no quantification of their use in humans has been carried out. Following a human intervention study with two mycotoxins, namely DON and deoxynivalenol-3-glucoside (DON3G), the renal excretion of these compounds, including their phase II metabolites, was analysed. The purpose was to develop biokinetic models that can be used to determine: (1) the preferred (set of) urinary biomarker(s), (2) the preferred urinary collection period, and (3) a method to estimate the dietary exposure to these mycotoxins. Twenty adult volunteers were restricted in consuming cereals and cereal-based foods for 4 days. At day 3, a single dose of 1 µg/kg body weight of DON or DON3G was orally administered to 16 volunteers; 4 volunteers served as control. All individual urine discharges were collected during 24 h after administration. The metabolism and renal excretion could be described by a biokinetic model using three physiological compartments (gastrointestinal tract, liver, and kidneys). Kinetic analysis revealed a complete recovery of the renal excretion of total DON (mainly DON and its glucuronides) within 24 h after administration of DON or DON3G. The so-called ‘reverse dosimetry’ factor was used to determine the preferred (set of) biomarker(s) and to estimate the dietary intake of the parent compounds in the future. The fact that DON3G was absorbed and mainly excreted as DON and its glucuronides confirms that DON3G (as well as other modified forms) should be taken into account in the exposure and risk assessment of this group of mycotoxins. Full article
(This article belongs to the Special Issue Mycotoxin Biomarkers of Exposure)
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Open AccessReview
Botulinum Toxin a Valuable Prophylactic Agent for Migraines and a Possible Future Option for the Prevention of Hormonal Variations-Triggered Migraines
Received: 30 June 2019 / Revised: 2 August 2019 / Accepted: 5 August 2019 / Published: 8 August 2019
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Abstract
Background: In 1989, Botulinum toxin (BoNT) was accepted by the FDA for the management of some ophthalmic disorders. Although it was initially considered a lethal toxin, in recent times, Botulinum toxin A (BoNT-A), which is the more used serotype, has expanded to cover [...] Read more.
Background: In 1989, Botulinum toxin (BoNT) was accepted by the FDA for the management of some ophthalmic disorders. Although it was initially considered a lethal toxin, in recent times, Botulinum toxin A (BoNT-A), which is the more used serotype, has expanded to cover different clinical conditions, primarily characterized by neuropathic pain, including migraines and headaches. Evidence suggests that migraines are influenced by hormonal factors, particularly by estrogen levels, but very few studies have investigated the prevalence and management strategies for migraines according to the hormonal status. The effects of several therapeutic regimens on migraines have been investigated, but the medications used varied widely in proven efficacies and mechanisms of action. BoNT-A is increasingly used in the management of migraine and several placebo-controlled trials of episodic and chronic migraine are currently underway. This paper is a review of the recently published data concerning the administration of BoNT-A in the prevention of chronic migraines. Considering the lack of population-based studies about the effectiveness of BoNT-A in the alleviation of premenstrual and perimenopausal migraines, this study proposes a new perspective of the therapeutic approach of migraine syndrome associated with menopausal transition and the premenstrual period. Methods: We selected the reviewed papers from CrossRef, PubMed, Medline, and GoogleScholar, and a total of 21 studies met our inclusion criteria. Results: To date, no specific preventive measures have been recommended for menopausal women with migraines. BoNT-A often reduces the frequency and intensity of migraine attacks per month; the treatment is well tolerated and does not exhibit a significantly higher rate of treatment-related side effects. No population-based studies were conducted in order to highlight the role of BoNT-A in menopause-related migraines, neither in menstrual migraines. Conclusion: There is a need for further research in order to quantify the real burden of menstrual and perimenopausal migraines and to clarify if BoNT-A could be used in the treatment of refractory postmenopausal and premenstrual migraines. Full article
(This article belongs to the Special Issue Botulinum Toxin for Neuropathic Pain Treatment)
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Open AccessArticle
Camelid VHHs Fused to Human Fc Fragments Provide Long Term Protection Against Botulinum Neurotoxin A in Mice
Received: 11 July 2019 / Revised: 29 July 2019 / Accepted: 2 August 2019 / Published: 7 August 2019
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Abstract
The bacterium Clostridium botulinum is the causative agent of botulism—a severe intoxication caused by botulinum neurotoxin (BoNT) and characterized by damage to the nervous system. In an effort to develop novel C. botulinum immunotherapeutics, camelid single-domain antibodies (sdAbs, VHHs, or nanobodies) could be [...] Read more.
The bacterium Clostridium botulinum is the causative agent of botulism—a severe intoxication caused by botulinum neurotoxin (BoNT) and characterized by damage to the nervous system. In an effort to develop novel C. botulinum immunotherapeutics, camelid single-domain antibodies (sdAbs, VHHs, or nanobodies) could be used due to their unique structure and characteristics. In this study, VHHs were produced using phage display technology. A total of 15 different monoclonal VHHs were selected based on their comlementarity-determining region 3 (CDR3) sequences. Different toxin lethal dose (LD50) challenges with each selected phage clone were conducted in vivo to check their neutralizing potency. We demonstrated that modification of neutralizing VHHs with a human immunoglobulin G (IgG)1 Fc (fragment crystallizable) fragment (fusionbody, VHH-Fc) significantly increased the circulation time in the blood (up to 14 days). At the same time, VHH-Fc showed the protective activity 1000 times higher than monomeric form when challenged with 5 LD50. Moreover, VHH-Fcs remained protective even 14 days after antibody administration. These results indicate that this VHH-Fc could be used as an effective long term antitoxin protection against botulinum type A. Full article
(This article belongs to the Section Bacterial Toxins)
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Open AccessArticle
Investigation of Zearalenone Adsorption and Biotransformation by Microorganisms Cultured under Cellular Stress Conditions
Received: 28 June 2019 / Revised: 3 August 2019 / Accepted: 5 August 2019 / Published: 7 August 2019
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Abstract
The zearalenone binding and metabolization ability of probiotic microorganisms, such as lactic acid bacteria, Lactobacillus paracasei, Lactococcus lactis, and yeast Saccharomyces cerevisiae, isolated from food products, were examined. Moreover, the influence of cellular stress (induced by silver nanoparticles) and lyophilization on the [...] Read more.
The zearalenone binding and metabolization ability of probiotic microorganisms, such as lactic acid bacteria, Lactobacillus paracasei, Lactococcus lactis, and yeast Saccharomyces cerevisiae, isolated from food products, were examined. Moreover, the influence of cellular stress (induced by silver nanoparticles) and lyophilization on the effectiveness of tested microorganisms was also investigated. The concentration of zearalenone after a certain time of incubation with microorganisms was determined using high-performance liquid chromatography. The maximum sorption effectiveness for L. paracasei, L. lactis, and S. cerevisiae cultured in non-stress conditions was 53.3, 41.0, and 36.5%, respectively. At the same time for the same microorganisms cultured at cellular stress conditions, the maximum sorption effectiveness was improved to 55.3, 47.4, and 57.0%, respectively. Also, the effect of culture conditions on the morphology of the cells and its metabolism was examined using microscopic technique and matrix-assisted laser desorption ionization-time of flight mass spectrometry, respectively. Full article
(This article belongs to the collection Fusarium Toxins – Relevance for Human and Animal Health)
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Open AccessArticle
Insight into the Structural Dynamics of the Lysenin During Prepore-to-Pore Transition Using Hydrogen–Deuterium Exchange Mass Spectrometry
Received: 18 July 2019 / Revised: 31 July 2019 / Accepted: 2 August 2019 / Published: 7 August 2019
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Abstract
Lysenin is a pore-forming toxin of the aerolysin family, which is derived from coelomic fluid of the earthworm Eisenia fetida. Upon binding to sphingomyelin (SM)-containing membranes, lysenin undergoes a series of structural changes promoting the conversion of water-soluble monomers into oligomers, leading [...] Read more.
Lysenin is a pore-forming toxin of the aerolysin family, which is derived from coelomic fluid of the earthworm Eisenia fetida. Upon binding to sphingomyelin (SM)-containing membranes, lysenin undergoes a series of structural changes promoting the conversion of water-soluble monomers into oligomers, leading to its insertion into the membrane and the formation of a lytic β-barrel pore. The soluble monomer and transmembrane pore structures were recently described, but the underlying structural details of oligomerization remain unclear. To investigate the molecular mechanisms controlling the conformational rearrangements accompanying pore formation, we compared the hydrogen–deuterium exchange pattern between lyseninWT and its mutant lyseninV88C/Y131C. This mutation arrests lysenin oligomers in the prepore state at the membrane surface and does not affect the structural dynamics of the water-soluble form of lysenin. In contrast, membrane-bound lyseninV88C/Y131C exhibited increased structural stabilization, especially within the twisted β-sheet of the N-terminal domain. We demonstrated that the structural stabilization of the lysenin prepore started at the site of lysenin’s initial interaction with the lipid membrane and was transmitted to the twisted β-sheet of the N-terminal domain, and that lyseninV88C/Y131C was arrested in this conformation. In lyseninWT, stabilization of these regions drove the conformational changes necessary for pore formation. Full article
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Open AccessArticle
Evolution of Asian Corn Borer Resistance to Bt Toxins Used Singly or in Pairs
Received: 1 July 2019 / Revised: 24 July 2019 / Accepted: 1 August 2019 / Published: 6 August 2019
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Abstract
Transgenic crops producing insecticidal proteins from Bacillus thuringiensis (Bt) have revolutionized pest control, but the benefits of this approach have been reduced by the evolution of resistance in pests. The widely adopted ’pyramid strategy’ for delaying resistance entails transgenic crops producing two or [...] Read more.
Transgenic crops producing insecticidal proteins from Bacillus thuringiensis (Bt) have revolutionized pest control, but the benefits of this approach have been reduced by the evolution of resistance in pests. The widely adopted ’pyramid strategy’ for delaying resistance entails transgenic crops producing two or more distinct toxins that kill the same pest. The limited experimental evidence supporting this strategy comes primarily from a model system under ideal conditions. Here we tested the pyramid strategy under nearly worst-case conditions, including some cross-resistance between the toxins in the pyramid. In a laboratory selection experiment with an artificial diet, we used Bt toxins Cry1Ab, Cry1F, and Cry1Ie singly or in pairs against Ostrinia furnacalis, one of the most destructive pests of corn in Asia. Under the conditions evaluated, pairs of toxins did not consistently delay the evolution of resistance relative to single toxins. Full article
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Open AccessReview
Biological Control of Citrus Postharvest Phytopathogens
Received: 11 July 2019 / Revised: 24 July 2019 / Accepted: 25 July 2019 / Published: 6 August 2019
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Abstract
Citrus are vulnerable to the postharvest decay caused by Penicillium digitatum, Penicillium italicum, and Geotrichum citri-aurantii, which are responsible for the green mold, blue mold, and sour rot post-harvest disease, respectively. The widespread economic losses in citriculture caused by these [...] Read more.
Citrus are vulnerable to the postharvest decay caused by Penicillium digitatum, Penicillium italicum, and Geotrichum citri-aurantii, which are responsible for the green mold, blue mold, and sour rot post-harvest disease, respectively. The widespread economic losses in citriculture caused by these phytopathogens are minimized with the use of synthetic fungicides such as imazalil, thiabendazole, pyrimethanil, and fludioxonil, which are mainly employed as control agents and may have harmful effects on human health and environment. To date, numerous non-chemical postharvest treatments have been investigated for the control of these pathogens. Several studies demonstrated that biological control using microbial antagonists and natural products can be effective in controlling postharvest diseases in citrus, as well as the most used commercial fungicides. Therefore, microbial agents represent a considerably safer and low toxicity alternative to synthetic fungicides. In the present review, these biological control strategies as alternative to the chemical fungicides are summarized here and new challenges regarding the development of shelf-stable formulated biocontrol products are also discussed. Full article
(This article belongs to the Special Issue Novel Approaches to Minimising Mycotoxin Contamination)
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Open AccessReview
Mechanisms of Botulinum Toxin Type A Action on Pain
Received: 14 July 2019 / Revised: 26 July 2019 / Accepted: 29 July 2019 / Published: 5 August 2019
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Abstract
Already a well-established treatment for different autonomic and movement disorders, the use of botulinum toxin type A (BoNT/A) in pain conditions is now continuously expanding. Currently, the only approved use of BoNT/A in relation to pain is the treatment of chronic migraines. However, [...] Read more.
Already a well-established treatment for different autonomic and movement disorders, the use of botulinum toxin type A (BoNT/A) in pain conditions is now continuously expanding. Currently, the only approved use of BoNT/A in relation to pain is the treatment of chronic migraines. However, controlled clinical studies show promising results in neuropathic and other chronic pain disorders. In comparison with other conventional and non-conventional analgesic drugs, the greatest advantages of BoNT/A use are its sustained effect after a single application and its safety. Its efficacy in certain therapy-resistant pain conditions is of special importance. Novel results in recent years has led to a better understanding of its actions, although further experimental and clinical research is warranted. Here, we summarize the effects contributing to these advantageous properties of BoNT/A in pain therapy, specific actions along the nociceptive pathway, consequences of its central activities, the molecular mechanisms of actions in neurons, and general pharmacokinetic parameters. Full article
(This article belongs to the Special Issue Botulinum Toxin for Neuropathic Pain Treatment)
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Open AccessArticle
Quercetin-3-Rutinoside Blocks the Disassembly of Cholera Toxin by Protein Disulfide Isomerase
Received: 5 July 2019 / Revised: 24 July 2019 / Accepted: 2 August 2019 / Published: 4 August 2019
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Abstract
Protein disulfide isomerase (PDI) is mainly located in the endoplasmic reticulum (ER) but is also secreted into the bloodstream where its oxidoreductase activity is involved with thrombus formation. Quercetin-3-rutinoside (Q3R) blocks this activity, but its inhibitory mechanism against PDI is not fully understood. [...] Read more.
Protein disulfide isomerase (PDI) is mainly located in the endoplasmic reticulum (ER) but is also secreted into the bloodstream where its oxidoreductase activity is involved with thrombus formation. Quercetin-3-rutinoside (Q3R) blocks this activity, but its inhibitory mechanism against PDI is not fully understood. Here, we examined the potential inhibitory effect of Q3R on another process that requires PDI: disassembly of the multimeric cholera toxin (CT). In the ER, PDI physically displaces the reduced CTA1 subunit from its non-covalent assembly in the CT holotoxin. This is followed by CTA1 dislocation from the ER to the cytosol where the toxin interacts with its G protein target for a cytopathic effect. Q3R blocked the conformational change in PDI that accompanies its binding to CTA1, which, in turn, prevented PDI from displacing CTA1 from its holotoxin and generated a toxin-resistant phenotype. Other steps of the CT intoxication process were not affected by Q3R, including PDI binding to CTA1 and CT reduction by PDI. Additional experiments with the B chain of ricin toxin found that Q3R could also disrupt PDI function through the loss of substrate binding. Q3R can thus inhibit PDI function through distinct mechanisms in a substrate-dependent manner. Full article
(This article belongs to the Section Bacterial Toxins)
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Open AccessEditorial
Staphylococcus aureus Toxins: Armaments for a Significant Pathogen
Received: 30 July 2019 / Accepted: 1 August 2019 / Published: 3 August 2019
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Abstract
Staphylococcus species are common inhabitants of humans and other animals [...] Full article
(This article belongs to the Special Issue Staphylococcus aureus Toxins) Printed Edition available
Open AccessArticle
Diagnosing Microcystin Intoxication of Canines: Clinicopathological Indications, Pathological Characteristics, and Analytical Detection in Postmortem and Antemortem Samples
Received: 13 June 2019 / Revised: 28 July 2019 / Accepted: 1 August 2019 / Published: 3 August 2019
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Abstract
In the summer of 2018, six dogs exposed to a harmful algal bloom (HAB) of Microcystis in Martin County Florida (USA) developed clinicopathological signs of microcystin (MC) intoxication (i.e., acute vomiting, diarrhea, severe thrombocytopenia, elevated alanine aminotransferase, hemorrhage). Successful supportive veterinary care was [...] Read more.
In the summer of 2018, six dogs exposed to a harmful algal bloom (HAB) of Microcystis in Martin County Florida (USA) developed clinicopathological signs of microcystin (MC) intoxication (i.e., acute vomiting, diarrhea, severe thrombocytopenia, elevated alanine aminotransferase, hemorrhage). Successful supportive veterinary care was provided and led to survival of all but one patient. Confirmation of MC intoxication was made through interpretation of clinicopathological abnormalities, pathological examination of tissues, microscopy (vomitus), and analytical MC testing of antemortem/postmortem samples (vomitus, blood, urine, bile, liver, kidney, hair). Gross and microscopic examination of the deceased patient confirmed massive hepatic necrosis, mild multifocal renal tubular necrosis, and hemorrhage within multiple organ systems. Microscopy of a vomitus sample confirmed the presence of Microcystis. Three analytical MC testing approaches were used, including the MMPB (2-methyl-3-methoxy-4-phenylbutyric acid) technique, targeted congener analysis (e.g., liquid chromatography tandem-mass spectrometry of MC-LR), and enzyme-linked immunosorbent assay (ELISA). Total Adda MCs (as MMPB) were confirmed in the liver, bile, kidney, urine, and blood of the deceased dog. Urinalysis (MMPB) of one surviving dog showed a high level of MCs (32,000 ng mL−1) 1-day post exposure, with MCs detectable >2 months post exposure. Furthermore, hair from a surviving dog was positive for MMPB, illustrating another testable route of MC elimination in canines. The described cases represent the first use of urine as an antemortem, non-invasive specimen to diagnose microcystin toxicosis. Antemortem diagnostic testing to confirm MC intoxication cases, whether acute or chronic, is crucial for providing optimal supportive care and mitigating MC exposure. Full article
(This article belongs to the Special Issue Freshwater Algal Toxins: Monitoring and Toxicity Profile)
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Open AccessArticle
Lack of Toxic Interaction between Fusariotoxins in Broiler Chickens Fed throughout Their Life at the Highest Level Tolerated in the European Union
Received: 26 June 2019 / Revised: 15 July 2019 / Accepted: 23 July 2019 / Published: 2 August 2019
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Abstract
Fusarium mycotoxins (FUS) occur frequently in poultry diets, and regulatory limits are laid down in several countries. However, the limits were established for exposure to a single mycotoxin, whereas multiple contamination is more realistic, and different studies have demonstrated that it is not [...] Read more.
Fusarium mycotoxins (FUS) occur frequently in poultry diets, and regulatory limits are laid down in several countries. However, the limits were established for exposure to a single mycotoxin, whereas multiple contamination is more realistic, and different studies have demonstrated that it is not possible to predict interactions between mycotoxins. The purpose of this study was thus to compare the toxic effect of deoxynivalenol (DON), fumonisins (FB) and zearalenone (ZON), alone and in combination on broiler chickens, at the maximum tolerated level established by the EU for poultry feed. Experimental corn-soybean diets incorporated ground cultured toxigenic Fusarium strains. One feed was formulated for chickens 0 to 10 days old and another for chickens 11 to 35 days old. The control diets were mycotoxin free, the DON diets contained 5 mg DON/kg, the FB diet contained 20 mg FB1 + FB2/kg, and the ZON diet contained 0.5 mg ZON/kg. The DONFBZON diet contained 5, 20, and 0.5 mg/kg of DON, FB1 + FB2, and ZON, respectively. Diets were distributed ad libitum to 70 broilers (male Ross PM3) separated into five groups of 14 chickens each reared in individual cages from one to 35 days of age. On day 35, after a starvation period of 8 h, a blood sample was collected, and all the animals were killed and autopsied. No difference between groups that could be attributed to FUS was observed in performances, the relative weight of organs, biochemistry, histopathology, intestinal morphometry, variables of oxidative damage, and markers of testicle toxicity. A significant increase in sphinganine and in the sphinganine to sphingosine ratio was observed in broilers fed FB. Taken together, these results suggest that the regulatory guidelines established for single contamination of broiler chickens fed with DON, FB, and ZON can also be used in the case of multiple contamination with these toxins. Full article
(This article belongs to the Special Issue Dietary Mycotoxin Exposure to Livestock and Poultry)
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Open AccessArticle
Indoxyl Sulfate Stimulates Angiogenesis by Regulating Reactive Oxygen Species Production via CYP1B1
Received: 14 June 2019 / Revised: 22 July 2019 / Accepted: 29 July 2019 / Published: 2 August 2019
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Abstract
Indoxyl sulfate (IS) is an accumulative protein-bound uremic toxin found in patients with kidney disease. It is reported that IS impairs the vascular endothelium, but a comprehensive overview of all mechanisms active in IS-injury currently remains lacking. Here we performed RNA sequencing in [...] Read more.
Indoxyl sulfate (IS) is an accumulative protein-bound uremic toxin found in patients with kidney disease. It is reported that IS impairs the vascular endothelium, but a comprehensive overview of all mechanisms active in IS-injury currently remains lacking. Here we performed RNA sequencing in human umbilical vein endothelial cells (HUVECs) after IS or control medium treatment and identified 1293 genes that were affected in a IS-induced response. Gene enrichment analysis highlighted pathways involved in altered vascular formation and cell metabolism. We confirmed these transcriptome profiles at the functional level by demonstrating decreased viability and increased cell senescence in response to IS treatment. In line with the additional pathways highlighted by the transcriptome analysis, we further could demonstrate that IS exposure of HUVECs promoted tubule formation as shown by the increase in total tubule length in a 3D HUVECs/pericytes co-culture assay. Notably, the pro-angiogenic response of IS and increased ROS production were abolished when CYP1B1, one of the main target genes that was highly upregulated by IS, was silenced. This observation indicates IS-induced ROS in endothelial cells is CYP1B1-dependent. Taken together, our findings demonstrate that IS promotes angiogenesis and CYP1B1 is an important factor in IS-activated angiogenic response. Full article
(This article belongs to the Special Issue The Endothelial Effects of Uremic Toxins)
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Open AccessArticle
Association of Long-Term Treatment by Botulinum Neurotoxins and Occupational Therapy with Subjective Physical Status in Patients with Post-Stroke Hemiplegia
Received: 19 June 2019 / Revised: 30 July 2019 / Accepted: 31 July 2019 / Published: 2 August 2019
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Abstract
The short-term effects of botulinum toxin type A (BoNT-A) treatment in stroke patients with upper extremity are well established. This study examined the association between the recovery of motor function of the upper extremity with subjective physical symptoms in outpatients receiving long-term BoNT-A [...] Read more.
The short-term effects of botulinum toxin type A (BoNT-A) treatment in stroke patients with upper extremity are well established. This study examined the association between the recovery of motor function of the upper extremity with subjective physical symptoms in outpatients receiving long-term BoNT-A and occupational therapy following stroke. We also investigated the expectations of patients who elected to continue treatment. Forty-seven patients (23 men and 24 women) aged 61 years received BoNT-A treatment more than 20 times. The subjective physical status was analyzed by using the visual analogue scale score through an eight-item questionnaire. Recovery of motor function in the upper extremity was detected by calculating the change (delta) in Fugl–Mayer Assessment (FMA), and ordinal logistic modeling analysis was used to determine the association between the delta-FMA score and the subjective level of agreement for each item. When the ordinal logistic modeling fit was statistically significant, results were interpreted as having logistic probability. The logistic curves discriminating one point (strongly disagree) from five points (strongly agree) were fit in a stepwise fashion. This study suggests that patients receiving long-term BoNT-A treatment and occupational therapy experienced an increased upper extremity mitigation and decreased insomnia after injection, regardless of the recovery of motor function. Full article
(This article belongs to the Special Issue Botulinum Toxin: The Role in Neuro-Rehabilitation)
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Open AccessArticle
Analysis of Mycotoxins Contamination in Poultry Feeds Manufactured in Selected Provinces of South Africa Using UHPLC-MS/MS
Received: 12 June 2019 / Revised: 4 July 2019 / Accepted: 6 July 2019 / Published: 2 August 2019
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Abstract
A total of 105 different types of poultry feed samples from South Africa were simultaneously analysed for the presence of 16 mycotoxins using ultra-high-performance liquid chromatography coupled to a triple quadrupole mass spectrometer (UHPLC-MS/MS). The data revealed the presence of 16 mycotoxins in [...] Read more.
A total of 105 different types of poultry feed samples from South Africa were simultaneously analysed for the presence of 16 mycotoxins using ultra-high-performance liquid chromatography coupled to a triple quadrupole mass spectrometer (UHPLC-MS/MS). The data revealed the presence of 16 mycotoxins in the various poultry feed samples. Fumonisin B1 (FB1) was the most dominant recovered from 100% of samples analysed at concentrations ranging between 38.7 and 7125.3 µg/kg. This was followed by zearalenone (ZEN) (range: 0.1–429 µg/kg) and deoxynivalenol (DON) (range: 2.5–154 µg/kg). Samples were also found to be contaminated with fumonisin B2 (FB2) (range: 0.7–125.1 µg/kg), fumonisin B3 (FB3) (range: 0.1–125.1 µg/kg), α-zearalenol (α-ZEL) (range: 0.6–20 µg/kg ), β-zearalenol (β-ZEL) (range: 0.2–22.1 µg/kg), 3-acetyldeoxynivalenol (3-ADON) (range: 0.1–12.9 µg/kg) and 15-acetyldeoxynivalenol (15-ADON) (range: 1.7–41.9 µg/kg). Alternaria mycotoxin, i.e., Alternariol monomethyl ether (AME) was recovered in 100% of samples at concentrations that ranged from 0.3–155.5 µg/kg. Aflatoxins (AFs) had an incidence rate of 92% with generally low concentration levels ranging from 0.1–3.7 µg/kg. Apart from these metabolites, 2 type A trichothecenes (THs), i.e., HT-2 toxin (HT-2) (range: 0.2–5.9 µg/kg) and T-2 toxin (T-2) (range: 0.1–15.3 µg/kg) were also detected. Mycotoxin contamination in South African poultry feed constitutes a concern as correspondingly high contamination levels, such as those observed herein are likely to affect birds, which can be accompanied by severe health implications, thus compromising animal productivity in the country. Such exposures, primarily to more than one mycotoxin concurrently, may elicit noticeable synergistic and or additive effects on poultry birds. Full article
(This article belongs to the Special Issue Fungal Growth and Mycotoxins: Challenges for developing countries)
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