Ameliorative effects of Lactobacillus against Aflatoxin B1 / Efeitos benéficos do Lactobacillus contra a Aflatoxin B1

Abstract Lactobacilli are probiotics with Aflatoxin (AF) detoxification ability, found in fermented products, GIT of animals and environment. Purpose of this study was to investigate the ability of broiler isolates of Lactobacillus against Aflatoxin B1 (AFB1). For this purpose, 5 isolates of Lactobacillus from broiler gut were incubated with 100 ppb AFB1 in aqueous environment and effect of different parameters (cell fractions, time, temperature, pH) on detoxification was determined by HPLC. The ameliorative effect of Lactobacillus salivarius (LS) against AFB1 was studied in broiler. The results revealed that LS (CR. 4) showed the best results (in vitro) as compared to other isolates (L. salivarius (CR. 3, CR, 4), L. agilis (CE. 2.1, CE. 3.1) and L. crispatus (CE. 28). Cell debris of CR. 4 showed significantly higher detoxification (P<0.05). Maximum amount of AFB1 was detoxified at 30°C (97%), pH 4.0 (99%) and 6 h (99.97%). In vivo study showed that AFB1 decreased weight gain (1,269 ± 0.04 gm/ bird), feed consumed (2,161 ± 0.08 gm/ bird), serum total protein (2.42 ± 0.34 gm/ dl), serum albumin (0.5 ± 0.2 2 gm/dl) and antibody titer (4.2 ± 0.83). Liver function enzymes were found (alanine transaminase (ALT): 32 ± 10.7 U/L) and aspartate transaminase (AST): 314.8 ± 27 U/L) elevated in AFB1 fed broilers. Treatment with 1% LS not only decreased the toxic effects of AFB1 (group D) but also improved the overall health of broilers due to its probiotic effects (p<0.05) as compared to control negative (group A). The detoxification ability of LS was better than commercial binder (CB) (0.2% Protmyc). It was concluded that detoxification of AFB1 by Lactobacillus was strain, temperature, pH and time dependent. LS has detoxification ability against AFB1 in vivo.

Resumo Os lactobacilos são probióticos com capacidade de desintoxicação da Aflatoxina (AF), encontrados em produtos fermentados, TGI de animais e meio ambiente. O objetivo deste estudo foi investigar a capacidade de isolados de frango de corte de Lactobacillus contra a Aflatoxina B1 (AFB1). Para tanto, 5 isolados de Lactobacillus de intestino de frango foram incubados com 100 ppb AFB1 em meio aquoso, e o efeito de diferentes parâmetros (frações celulares, tempo, temperatura, pH) na desintoxicação foi determinado por CLAE. O efeito melhorador de Lactobacillus salivarius (LS) contra AFB1 foi estudado em frangos de corte. Os resultados revelaram que LS (CR. 4) apresentou os melhores resultados (in vitro) em comparação com outros isolados [L. salivarius (CR. 3, CR. 4), L. agilis (CE. 2.1, CE. 3.1) e L. crispatus (CE. 28)]. Detritos celulares de CR. 4 mostraram desintoxicação significativamente maior (P < 0.05). A quantidade máxima de AFB1 foi desintoxicada a 30 °C (97%), pH 4.0 (99%) e 6 h (99,97%). O estudo in vivo mostrou que AFB1 diminuiu o ganho de peso (1,269 ± 0.04 g / ave), alimento consumido (2,161 ± 0.08 g / ave), proteína total sérica (2.42 ± 0.34 g / dl), albumina sérica (0.5 ± 0.22 gm / dl) e título de anticorpo (4.2 ± 0.83). As enzimas da função hepática foram encontradas (alanina transaminase (ALT): 32 ± 10.7 U / L) e aspartato transaminase (AST): 314.8 ± 27 U / L) elevadas em AFB1 alimentados com frangos. O tratamento com 1% LS não só diminuiu os efeitos tóxicos de AFB1 (grupo D), mas também melhorou a saúde geral dos frangos devido aos seus efeitos probióticos (p < 0.05) em comparação com o controle negativo (grupo A). A capacidade de desintoxicação do LS foi melhor do que o aglutinante comercial (CB) (0.2% Protmyc). Concluiu-se que a desintoxicação de AFB1 por Lactobacillus foi dependente da cepa, temperatura, pH e tempo. LS tem capacidade de desintoxicação contra AFB1 in vivo.

Ameliorative effects of Lactobacillus against Aflatoxin B1

Abstract Lactobacilli are probiotics with Aflatoxin (AF) detoxification ability, found in fermented products, GIT of animals and environment. Purpose of this study was to investigate the ability of broiler isolates of Lactobacillus against Aflatoxin B1 (AFB1). For this purpose, 5 isolates of Lactobacillus from broiler gut were incubated with 100 ppb AFB1 in aqueous environment and effect of different parameters (cell fractions, time, temperature, pH) on detoxification was determined by HPLC. The ameliorative effect of Lactobacillus salivarius (LS) against AFB1 was studied in broiler. The results revealed that LS (CR. 4) showed the best results (in vitro) as compared to other isolates (L. salivarius (CR. 3, CR, 4), L. agilis (CE. 2.1, CE. 3.1) and L. crispatus (CE. 28). Cell debris of CR. 4 showed significantly higher detoxification (P 0.05). Maximum amount of AFB1 was detoxified at 30°C (97%), pH 4.0 (99%) and 6 h (99.97%). In vivo study showed that AFB1 decreased weight gain (1,269 ± 0.04 gm/ bird), feed consumed (2,161 ± 0.08 gm/ bird), serum total protein (2.42 ± 0.34 gm/ dl), serum albumin (0.5 ± 0.2 2 gm/dl) and antibody titer (4.2 ± 0.83). Liver function enzymes were found (alanine transaminase (ALT): 32 ± 10.7 U/L) and aspartate transaminase (AST): 314.8 ± 27 U/L) elevated in AFB1 fed broilers. Treatment with 1% LS not only decreased the toxic effects of AFB1 (group D) but also improved the overall health of broilers due to its probiotic effects (p 0.05) as compared to control negative (group A). The detoxification ability of LS was better than commercial binder (CB) (0.2% Protmyc). It was concluded that detoxification of AFB1 by Lactobacillus was strain, temperature, pH and time dependent. LS has detoxification ability against AFB1 in vivo.

Resumo Os lactobacilos são probióticos com capacidade de desintoxicação da Aflatoxina (AF), encontrados em produtos fermentados, TGI de animais e meio ambiente. O objetivo deste estudo foi investigar a capacidade de isolados de frango de corte de Lactobacillus contra a Aflatoxina B1 (AFB1). Para tanto, 5 isolados de Lactobacillus de intestino de frango foram incubados com 100 ppb AFB1 em meio aquoso, e o efeito de diferentes parâmetros (frações celulares, tempo, temperatura, pH) na desintoxicação foi determinado por CLAE. O efeito melhorador de Lactobacillus salivarius (LS) contra AFB1 foi estudado em frangos de corte. Os resultados revelaram que LS (CR. 4) apresentou os melhores resultados (in vitro) em comparação com outros isolados [L. salivarius (CR. 3, CR. 4), L. agilis (CE. 2.1, CE. 3.1) e L. crispatus (CE. 28)]. Detritos celulares de CR. 4 mostraram desintoxicação significativamente maior (P 0.05). A quantidade máxima de AFB1 foi desintoxicada a 30 °C (97%), pH 4.0 (99%) e 6 h (99,97%). O estudo in vivo mostrou que AFB1 diminuiu o ganho de peso (1,269 ± 0.04 g / ave), alimento consumido (2,161 ± 0.08 g / ave), proteína total sérica (2.42 ± 0.34 g / dl), albumina sérica (0.5 ± 0.22 gm / dl) e título de anticorpo (4.2 ± 0.83). As enzimas da função hepática foram encontradas (alanina transaminase (ALT): 32 ± 10.7 U / L) e aspartato transaminase (AST): 314.8 ± 27 U / L) elevadas em AFB1 alimentados com frangos. O tratamento com 1% LS não só diminuiu os efeitos tóxicos de AFB1 (grupo D), mas também melhorou a saúde geral dos frangos devido aos seus efeitos probióticos (p 0.05) em comparação com o controle negativo (grupo A). A capacidade de desintoxicação do LS foi melhor do que o aglutinante comercial (CB) (0.2% Protmyc). Concluiu-se que a desintoxicação de AFB1 por Lactobacillus foi dependente da cepa, temperatura, pH e tempo. LS tem capacidade de desintoxicação contra AFB1 in vivo.

Determination of Knowledge Based on Aflatoxin Poisoning among Broiler Farmers in Nairobi City County, Kenya

Background: There is a scarcity of information concerning knowledge of aflatoxin contamination of feeds among farmers even in aflatoxin-prone regions in Kenya. Thus, knowledge of aflatoxins in feeds among poultry farmers is of paramount importance in designing plans to minimize risks of aflatoxin exposure. Therefore, this study sought to assess the Determinants of Knowledge on Aflatoxin Among Broiler farmers in Nairobi City County, Kenya. Methodology: The study utilized an analytical cross-sectional study design. A total of 240 farmers were sampled from a population of 600 farmers within Nairobi City County. A structured questionnaire was administered to farmers within Nairobi City County. SPSS version 26 was used to analyze the data descriptively. Results were presented in tables and figures. Ethical approval was sought from relevant authorities and parties before the commencement of the study. Results: Results from the study show that the majority of the farmers (58.2%) had knowledge of aflatoxin. There was a significant association (p<0.05) between the socio-demographic characteristics of farmers and knowledge of aflatoxin. Conclusion: The study concludes that the farmers had adequate knowledge of aflatoxin occurrence in feeds and methods to reduce the contamination. There is a need for continuous sensitization of farmers on aflatoxin, particularly on feed management practices by the Ministry of Agriculture and Ministry of Health Division of Public Health in Kenya.

Analysis of Aflatoxin Levels in Broiler Chicken Feed from Selected Farms in Nairobi City County, Kenya

Aflatoxin levels in animal feed should be observed from the farm to the table to ensure the safety of the feed to animals and humans. The contamination of cereals and other agricultural supplies used in animal feed production could happen in the farm in the pre-harvest phase or in the post-harvest phase. The study sought to determine Aflatoxin levels in broiler feed from selected farms in Nairobi City County. A total of 42 feed samples were collected. Samples were analyzed using the LCMS/MS technique. Results from the study show that Aflatoxin levels in broiler starter were; B1(17.26±3.07 ppb), B2 (2.44±0.84 ppb), G1 (8.87±2.41 ppb), G2 (0.9±0.44 ppb) and Total AF (29.47±6.13 ppb). Aflatoxin levels in broiler finisher were B1 (17.17±3.09 ppb), B2 (2.68±1.18 ppb), G1 (9.25±2.7 ppb), G2 (1±0.45 ppb) and Total AF (30.1±6.88 ppb). Results from analysis of feed samples showed that AFB1 levels in both broiler starter and broiler finisher were above the KEBS limit but were below the EAC, EU and WHO/FAO limit. Total Aflatoxin levels were above the KEBS limit but below the EAC limit. There is need to enhance the capacity of feed surveillance and monitoring in the country through various laboratory analysis techniques among various agencies in the feed value chain to ensure feed safety.

The synergism of hepatitis B and aflatoxicosis in hepatocellular carcinoma development: A mini-review

Hepatocellular carcinoma (HCC) is the twelfth most common cancer and the fifth leading cause of worldwide cancer related death. Chronic hepatitis B infection, caused by the hepatitis B virus (HBV) and exposure to aflatoxins is fundamental in the formation of HCC in developing countries. This review of scientific publications aims to establish the detrimental effects of aflatoxin-contaminated foods and highlights the correlation between aflatoxin and hepatitis B viral-associated hepatocellular carcinoma. Research has shown a significant increase in the occurrence of HCC in HBV-infected individuals exposed to fungal toxins. HBV demonstrates the ability to integrate and bind to p53 protein in the host DNA and propagate hepatocyte vulnerability through carcinogenic aflatoxin B1 (AFB1) damage. Although there has been clear evidence about the synergistic interaction of exposure to AFB1 and HBV infection in the induction of HCC, other literature has shown otherwise, mainly because incomplete and vague findings and hypotheses were made in regions where AFB1 and HBV pose a public health risk. Vaccination against hepatitis B and measures such as robust food safety systems to avoid hepatotoxicity and hepatocellular carcinogenesis induced by AFB1 is the most effective methods in the prevention of HCC induced by HBV and AFB1

Aflatoxin Occurrence, Food Regulations, Dietary Exposure, and Risk Assessment: A Mini Review from the Malaysian Perspective

@#Aflatoxins are ubiquitous and occur in food. Exposure to aflatoxins seriously impact the health of human and animal. It is concerning especially when aflatoxins are odourless, colourless, and tasteless that hardly be detected through naked eyes. Ingestion of aflatoxin-contaminated food contributes the major route of exposure. The present review is an update on the aflatoxin occurrence in food, aflatoxin regulations in food, and recent risk assessment of aflatoxin exposure in Malaysia. Peanuts and chili were more prone to aflatoxin contamination in Malaysia. The extreme weather experienced in Malaysia and global climatic change may worsen the aflatoxin contamination in food. The regulatory standards for aflatoxins imposed by Malaysia are less stringent than developed countries. The dietary exposure of aflatoxins among Malaysian was relatively high as compared with other Asia countries, ranging from 0.002 to 34.00 ng/kg body weight/day. Nonetheless, Malaysian population had low risk of aflatoxin-related liver cancer, with an estimated liver cancer risk of <1 cancer case/100,000 population/year.

Determination of aflatoxin M1 in breast milk and related factors

SUMMARY OBJECTIVE: Breastfeeding in women with aflatoxin M1 exposure may be a risk factor for the newborn. Thus, it is crucial to determine aflatoxin M1 levels in breast milk and raise mothers' awareness about nutrition in lactation and other periods. This study was carried out to determine aflatoxin M1 contamination in milk samples taken from mothers who gave birth. METHODS: The study was carried out in the postpartum department of Training and Research Hospital between December 31, 2018, and June 31, 2019, and 90 breastfeeding mothers were included in the study. RESULTS: A total of 75 (83.3%) of the examined samples were found positive. The mean aflatoxin M1 ratio in positive samples was 12.16 pg/mL (5.00-23.18 pg/mL). Mothers' consumption of processed food was associated with aflatoxin M1 levels (p=0.043). It was determined that the aflatoxin M1 levels of mothers who consumed processed food products 1 or 2 times a month were 3.22 times lower than those who consumed 1-2 times a week. CONCLUSIONS: This study emphasized the importance of monitoring aflatoxin M1 levels in breast milk for infant health. It is thought that nutrition education given to mothers during pregnancy will significantly impact aflatoxin M1 results. In addition, the dangers of mycotoxins in mother-infant nutrition should be emphasized regularly in health education.

Análise físico-química e atividades antifúngica e antioxidante do óleo essencial da flor de Curatella americana L / Physichochemical analysis, and antifungal and antioxidant activities of the essential oil of Curatella americana L. flower

Introdução: a espécie vegetal Curatella americana produz anualmente inflorescências com aroma adocicado rica em óleo essencial. Objetivo: avaliar as características físico-químicas, e atividades antifúngica e antioxidante do óleo essencial da flor de Curatella americana. Metodologia: as flores foram coletadas em quatro áreas de Cerrado no estado de Goiás; o rendimento de óleo essencial foi obtido através de hidrodestilação; as características físicas foram determinadas para densidade e solubilidade, a atividade antioxidante foi determinada pela redução do radical livre DPPH; a atividade antifúngica foi determinada por inibição das cepas de Candida, Sclerotinia sclerotiorum, Colletotrichum gloeosporioides e Aspergillus flavus. Resultados: o rendimento de óleo foi de 0,18%, densidade de 0,907 g mL-1, solubilidade positiva para EtOH 70%, atividade antioxidante de CI50 µL mL-1 1,95. Atividade de inibição fúngica apenas para Candida tropicalis na concentração de 8% com halo de antibiose de 10 mm. Sensibilidade discreta nas maiores concentrações de 25, 50 e 100 µL-1 para Aspergillus flavus e Sclerotinia sclerotiorum e baixa atividade de inibição para Colletotrichum gloeosporioides. Conclusão: o óleo essencial da flor de Curatella americana apresentou baixo rendimento, entretanto, alta eficiência na redução do radical livre DPPH. As atividades antifúngicas apresentaram bons resultados de inibição, entretanto, torna-se necessário a adição de outros óleos essenciais para aumento das taxas de inibição micelial.

Introduction: the plant species Curatella americana produces annual inflorescences with a sweet flavour rich in essential oil. Objective: to evaluate the physicochemical characteristics, antifungal and antioxidant activities of the essential oil of the Curatella americana flower. Methodology: the flowers were collected in four areas of Cerrado in the state of Goiás; the essential oil yield was obtained through hydrodistillation; the physical characteristics were determined for density and solubility, the antioxidant activity was determined by the reduction of the free radical DPPH; antifungal activity was determined by inhibiting the strains of Candida, Sclerotinia sclerotiorum, Colletotrichum gloeosporioides and Aspergillus flavus. Results: the oil yield was 0.18%, density 0.907 g mL-1, positive solubility for EtOH 70%, antioxidant activity of IC50 µL mL-1 1.95. Fungal inhibition activity only for Candida tropicalis at a concentration of 8% with a 10 mm antibiosis halo. Discrete sensitivity in the highest concentrations of 25, 50 and 100 µL-1 for Aspergillus flavus and Sclerotinia sclerotiorum and low inhibition activity for Colletotrichum gloeosporioides. Conclusion: The essential oil of the Curatella americana flower showed low yield, however, high efficiency in reducing DPPH free radical. Antifungal activities showed good inhibition results, however, it is necessary to add other essential oils to increase mycelial inhibition rates.

Aflatoxin levels and prevalence of TP53 aflatoxin-mutations in hepatocellular carcinomas in Mexico / Niveles de aflatoxina y prevalencia de mutaciones del gen TP53 por aflatoxina en carcinoma hepatocelular en México

Abstract: Objective: To determine the exposure to aflatoxin B1 (AFB1) in southern Mexico and the presence of the aflatoxin signature mutation in hepatocellular carcinoma (HCC) tissue from patients from a cancer referral center. Materials and methods: We estimated the prevalence and distribution of AFB1 in a representative sample of 100 women and men from Chiapas using the National Health and Nutrition Survey 2018-19. We also examined the presence of the aflatoxin signature mutation in codon 249 (R249S), and other relevant mutations of the TP53 gene in HCC tissue blocks from 24 women and 26 men treated in a national cancer referral center. Results: The prevalence of AFB1 in serum samples was 85.5% (95%CI 72.1-93.1) and the median AFB1 was 0.117 pg/µL (IQR, 0.050-0.350). We detected TP53 R249S in three of the 50 HCCs (6.0%) and observed four other G>T transversions potentially induced by AFB1. Conclusion: Our analysis provides evidence that AFB1 may have a relevant role on HCC etiology in Mexico.

Resumen: Objetivo: Determinar la exposición a aflatoxina_B1 (AFB1) en el sur de México y la presencia de la mutación característica de AFB1 en tejido de carcinoma hepatocelular (CHC) de pacientes de un centro oncológico. Material y métodos: Se estimó la prevalencia y distribución de AFB1 en una muestra representativa de 100 mujeres y hombres de Chiapas a partir de la Encuesta Nacional de Salud y Nutrición 2018-19. También se observó la presencia de la mutación característica de AFB1 en el codón 249 (R249S), y otras mutaciones relevantes del gen TP53 en bloques de tejido de CHC de 24 mujeres y 26 hombres estudiados en un centro de referencia nacional de oncología. Resultados: La prevalencia de AFB1 en las muestras de suero fue de 85.5% (IC95% 72.1-93.1) y la mediana de la concentración 0.117 pg/µL (IQR, 0.050-0.350). Se detectó TP53 R249S en tres de 50 casos de CHC (6.0%) y se observaron cuatro transversiones G>T potencialmente inducidas por AFB1. Conclusión: El presente análisis proporciona evidencia de que la AFB1 puede tener un papel relevante en la etiología del CHC en México.

The use of histológical parameters to assess intestinal and liver health on broilers challenged isolatedly and simultaneously with cyclopiazonic acid and aflatoxin / B1 El uso de parámetros histológicos para evaluar la salud intestinal y hepática en pollos de engorde desafiados de forma aislada y simultánea con ácido ciclopiazónico y aflatoxina B1

Abstract Mycotoxins contaminate agricultural commodities, which contaminates animals. These toxins can damage vital organs, such as the liver, as well as the epithelial tissue. Among these mycotoxins are aflatoxin B1 (AFB1) and cyclopiazonic acid (CPA), which can occur simultaneously in food. In broilers, mycotoxicosis has an economic impact due to several factors, such as low feed conversion rate, incidence of other diseases, and interference with reproductive capacity, all of which may lead to a public health problem. The aim of the present study was to histologically assess, through the I See Inside (ISI) method, harmful effects on broiler liver, duodenum, jejunum, and ileum in the presence of AFB1 and CPA isolatedly and simultaneously. Groups challenged with mycotoxins showed significant damage to both gut and liver fragments. All challenged-groups in all fragments impaired the parameters analyzed for intestinal epithelium. In the liver, AFB1 was predominantly harmful when the parameters were analyzed separately, but when analyzing the total ISI score, CPA was also found to be harmful to this organ. The other point analyzed was the great variation between the weights of the birds contaminated by mycotoxin while the negative control group presents a lesser variation.

Resumen Las micotoxinas contaminan los productos agrícolas, que a su vez contaminan a los animales. Estas toxinas pueden dañar órganos vitales, como el hígado y el tejido epitelial. Entre estas micotoxinas se encuentran la aflatoxina B1 (AFB1) y el ácido ciclopiazónico (CPA), que pueden hallarse simultáneamente en los alimentos. En los pollos de engorde, la micotoxicosis tiene un impacto económico debido a varios factores, como la baja tasa de conversión alimenticia, la incidencia de otras enfermedades y la interferencia de la capacidad reproductiva, que pueden llevar a un problema de salud pública. El objetivo de la presente investigación es la de evaluar histológicamente, a través del método "I See Inside" (ISI), los efectos nocivos sobre el hígado, duodeno, yeyuno e íleon de pollos de engorde en presencia de AFB1 y CPA de forma aislada y simultánea. Los grupos desafiados con micotoxinas presentaron un daño significativo tanto en el intestino como en los fragmentos del hígado. Todos los grupos tratados tuvieron alteraciones en los parámetros analizados para el epitelio intestinal. En el hígado, AFB1 fue predominantemente dañino cuando los parámetros se analizaron por separado, pero al examinar la puntuación ISI total, también se encontró que el CPA era perjudicial para este órgano. Otra cuestión que fue investigada fue la gran variación entre los pesos de las aves contaminadas por micotoxinas mientras el grupo de control negativo presentó una variación menor.

Factors contributing to urinary aflatoxin M1 occurrence among residents in Hulu Langat district, Malaysia

@#Introduction: Chronic exposure to aflatoxin can lead to complications such as liver failure and cancer. There are many factors that affect aflatoxin occurrence. This study aimed to assess the association between sociodemographic factors and the knowledge, attitude and practice towards aflatoxin with urinary aflatoxin M1 occurrence among residents in Hulu Langat district, Malaysia. Methods: This was a cross-sectional study conducted among healthy Malaysian adults aged 18 to 60 years residing in Hulu Langat district, Malaysia. Socio-demographic background and the knowledge, attitude and practice of respondents towards aflatoxin were assessed through questionnaires. Non-fasting urine sample (15 ml) was collected in the morning and urinary aflatoxin M1 level was quantified. Results: Of the 444 healthy Malaysian adults, 199 urine samples were detected with aflatoxin M1. From 37 positive samples with aflatoxin M1 level above detection limit (0.64 ng/ml), mean value was 1.23±0.91 ng/ml (range = 0.65-5.34 ng/ml). Urinary aflatoxin M1 occurrence was significantly different across ethnicity, age group, monthly household income, attitude and practice towards aflatoxin. Binomial logistic regression confirmed ethnicity and monthly household income as factors contributing to urinary aflatoxin M1 occurrence. Chinese were 3.20 times more likely to have aflatoxin exposure than non-Chinese. Detected urinary aflatoxin M1 was more common among household with a monthly income above RM1,500. Conclusion: The results provided an insight to explain the variation in aflatoxin occurrence among the population.

Rapid detection of aflatoxin B_1 in Chinese herbal medicines by indirect and direct competitive enzyme-linked immunosorbent assays: a comparative analysis / 中国中药杂志

The indirect competitive enzyme-linked immunosorbent assay(ic-ELISA) and direct competitive enzyme-linked immunosorbent assay(dc-ELISA) were performed for the rapid detection of aflatoxin B_1(AFB_1) in Astragali Radix, Zingiberis Rhizoma Recens, Jujubae Fructus, and Nelumbinis Semen with self-made antigens and antibodies. Different extraction methods were investigated to reduce the matrix effects of different medicinal parts in Chinese herbal medicines. The sensitivity of dc-ELISA method was improved by optimizing the molar ratio of AFB_1 to horseradish peroxidase(HRP). In this study, the sensitivity(IC_(50)) of ic-ELISA and dc-ELISA was 0.046 and 0.023 ng·mL~(-1), with the limit of detection(LOD) of 0.007 and 0.004 ng·mL~(-1), respectively. The detection time was 3 h and 50 min for ic-ELISA and dc-ELISA, respectively. The recovery rates were within the range of 62.96%-104.4%, with RSDs of less than 10%. Confirmed by LC-MS/MS, three positive samples of Nelumbinis Semen were detected from 53 samples. Two ELISA methods established in this study were accurate, rapid and sensitive, and can be used for rapid screening of AFB_1 in Chinese herbal medicines such as Astragali Radix, Zingiberis Rhizoma Recens, Jujubae Fructus, and Nelumbinis Semen. In addition, the advantages and limitations of the two methods were compared and discussed, which can provide a reference for the testing institutions to choose the proper method.

Analysis of Chemical Composition in Essential Oil of Zanthoxyli Pericarpium from Different Producing Areas and Investigation of Antifungal Effect of Zanthoxyli Pericarpium Essential Oil Solid Preparation / 中国实验方剂学杂志

Objective:To extract essential oil of Zanthoxyli Pericarpium， to prepare Zanthoxyli Pericarpium essential oil solid preparation and investigate its anti-fungal effect， in order to provide safe， green and efficient fungicide for the storage of Chinese herbal medicine and food. Method:The essential oil of Zanthoxyli Pericarpium was extracted by steam distillation method， gas chromatography-mass spectrometry （GC-MS） was adopted to analyze the chemical compositions and their relative contents in essential oil of Zanthoxyli Pericarpium from different producing areas， Agilent HP-5 capillary column was used for separation at programmed temperature （the initial temperature was 60 ℃， kept for 2 min， then increased to 280 ℃ by 10 ℃·min^-1， kept for 5 min）， the scanning range was <italic>m</italic>/<italic>z</italic> 35-590. Zanthoxyli Pericarpium essential oil solid preparation was prepared by nanomolecular sieve adsorption method， and its inhibitory effect on <italic>Aspergillus flavus</italic> and its conidia was investigated. Ultra-high performance liquid chromatography-fluorescence detector （UPLC-FLD） was used to analyze the inhibitory effect of Zanthoxyli Pericarpium essential oil solid preparation on aflatoxin under the conditions of excitation wavelength of 360 nm and emission wavelength of 440 nm. Result:The average extraction rate of essential oil in Zanthoxyli Pericarpium from four producing areas was 5.2%. （+）-Limonene， linalool and linalyl acetate were the main components of Zanthoxyli Pericarpium<italic> </italic>essential oil<italic> </italic>from different producing areas. When the volume fraction of essential oil in the solid preparation was 0.1%， the inhibition rate of the solid preparation on the conidia of <italic>A</italic>. <italic>flavus</italic> was （16.41±8.89）%. When the volume fraction of essential oil in the solid preparation was 0.2%， the inhibition rate for the growth of <italic>A</italic>. <italic>flavus</italic> was （8.11±2.70）%. When the volume fraction of essential oil in the solid preparation was 0.5%， the inhibition rate for the growth of <italic>A</italic>. <italic>flavus </italic>was （21.62±5.41）%， the inhibition rate for <italic>A</italic>. <italic>flavus</italic> conidia was （45.43±5.67）%， and the inhibition effect for the aflatoxin could reach （90.47±12.77）%. Conclusion:There are some differences in the chemical composition of essential oil of Zanthoxyli Pericarpium from different producing areas. Zanthoxyli Pericarpium<italic> </italic>essential oil has a certain inhibitory effect on the formation of <italic>A. flavus</italic> conidia and the production of aflatoxin B₁. It shows that Zanthoxyli Pericarpium essential oil can be developed into bacteriostatic preparation and used in the storage of Chinese medicinal materials and food.

Detección de aflatoxina M1 en muestras de leche cruda de vacas en tanques de enfriamiento en Boyacá (Colombia) / Detection of aflatoxin M1 in raw cow milk samples in cooling tanks in Boyacá (Colombia)

RESUMEN Una de las principales formas de contaminación de la leche con micotoxinas es el consumo de alimentos fermentados que se encuentran contaminados con mohos principalmente de Aspergillus spp., los cuales producen toxinas que pueden llegar a constituirse como un problema para la salud publica debido a su estabilidad térmica y química. El objetivo del presente trabajo fue detectar las concentraciones de aflatoxina M1 en muestras de leche de vacas en tanques de enfriamiento en cuatro municipios del departamento de Boyacá durante un año, determinando las variaciones de acuerdo con la temporada. Se realizó un estudio de corte longitudinal, descriptivo cuantitativo. Se seleccionaron aleatoriamente cuatro tanques de enfriamiento de cuatro municipios distintos del departamento; cada uno se muestreó dos veces al mes durante todo el período de estudio y se procesaron mediante metodología Charm Ez Lite . Se realizó un ANDEVA para determinar las diferencias estadísticas entre las concentraciones de la aflatoxina M1 por cada trimestre. Se determinaron diferencias estadísticas entre cada uno de los trimestres del estudio encontrando un porcentaje de positividad de 74,06% del total de muestras positivas en los trimestres de verano. 28,12% (108) de las muestras tomadas durante todo el estudio fueron positivas, con concentraciones de la toxina que oscilaron entre 0,5 y 2,0 μg/Kg de leche. Se determinó por primera vez en el departamento de Boyacá las concentraciones y variaciones estacionales de aflatoxina M1 en muestras de tanques de enfriamiento de leche, encontrando las mayores concentraciones y número de casos positivos de aflatoxina M1 en los meses de verano.

ABSTRACT One of the main forms of contamination of milk with mycotoxins is the consumption of fermented foods that are contaminated with mold, mainly Aspergillus spp, which produce toxins that can become a public health problem due to their thermal and chemical stability. The objective of the present work was to detect aflatoxin M1 concentrations in cows' milk samples in cooling tanks in four municipalities of the department of Boyacá for one year, determining the variations according to the season. A longitudinal, quantitative descriptive study was carried out, four cooling tanks from four different municipalities in the department were randomly selected, each tank, in each municipality, was sampled twice a month throughout the study period and processed using Charm methodology Ez Lite®, an ANDEVA was performed to determine the statistical differences between aflatoxin M1 concentrations for each quarter. Statistical differences were determined between each of the quarters of the study, finding a positivity percentage of 74.06% of the total positive samples in the summer quarters. 28.12% (108) of the samples taken throughout the study were positive, with toxin concentrations ranging between 0.5 and 2.0 μg/Kg of milk. Seasonal concentrations and variations of aflatoxin M1 in milk cooling tank samples were determined for the first time in the department of Boyacá, finding the highest concentrations and number of positive cases of aflatoxin M1 in the summer months.

Metabolic Syndrome Instead of Aflatoxin-Related TP53 R249S Mutation as a Hepatocellular Carcinoma Risk Factor

Abstract Background: Hepatocarcinogenesis has a variety of risk factors. In Mexico City, autopsies found 14% of hepatocellular carcinomas (HCCs) without cirrhosis. Objective: The objective of the study was to explore if HCCs carry the TP53 R249S mutation that has linked them to aflatoxin exposure and describe the associated risk factors. Methods: A retrospective review of consecutive cases of HCC was performed. Exposure to hepatotropic viruses, alcoholism, metabolic diseases, diabetes mellitus, and hypertension, as well as episodes of ascites, portal hypertension, and body mass index were retrieved. Slides were re-reviewed, macrodissected and DNA was extracted. TP53 exon 7 was amplified, purified, and used as a template for sequencing. Results: In 14 years, 74 HCCs were identified in 1863 (4%) consecutive liver biopsies. No data were available in five excluded patients; the rest was submitted to exon 7 screening. Patients had a median age of 62 years, and 46 (67%) were male. Stage 4 fibrosis was observed in 46 patients (67%) and their associated risk factors were hepatitis C virus (39%, 18/46), alcoholism (20%, 9/46), hepatitis B virus (2%, 1/46), and 18 were cryptogenic. Fibrosis stage 3 or lower was observed in 23 (33%) patients without demonstrated liver disease; 8/23 had diabetes and 6/23, systemic hypertension. Steatohepatitic variants of HCC were observed in 4 and in 5, the remnant liver had steatohepatitis. A 238-bp fragment was obtained in each tumor without the expected TP53 R249S mutation. Conclusions: There was no evidence of aflatoxin exposure in HCCs, with and without the known “classical” risk factors. One-third of non-cirrhotic HCCs had steatohepatitis or conditions associated to metabolic syndrome. (REV INVEST CLIN. 2020;72(5):316-22)

Biopanning the mimotopes of aflatoxin B1 and their immunogenicity / Panning de mimotopo aflatoxina B1 e sua imunogenicidade

Utilizando um anticorpo monoclonal contra a aflatoxina B1 (AFB1) como ligante, foi identificado um mimotopo específico de aflatoxina B1 após se realizarem quatro ciclos de seleção biológica de 7-peptídeos aleatórios em biblioteca de fago exibida. O mimotopo é denominado P10, e sua sequência de aminoácidos é YRRHEKD. O soro imunológico de ratos Balb/c imunizados com P10 foi especificamente ligado à aflatoxina B1-albumina, indicando que o anticorpo era específico ao AFB1. Esses resultados sugerem que é possível desenvolver a vacina baseada em mimotopo associado à toxina.(AU)

Physiochemical Screening of Carica papaya Leaves with Specific Reference to Their Pharmacognostical Evaluation

Carica papaya is made to develop pharmacognostical characters of leaf with their morphological, microscopical, and physical characters including histochemical analysis. Morphological evaluation as color, odor, taste, size, shape, surface, and powder microscopy of plant shows the presence of endosperm cell which is polygonal in shape and contains aleurone grains and oil droplet, cell of testa, yellow coloring matter, and starch grains. Quantitative leaf microscopy to determine palisade ratio, stomata index, and vein-islet number is carried out. Peels are removed mechanically through epidermal peeling off and stomatal index (SI) is calculated. The vein-islet number, vein termination number, and palisade ratio of lamina are determined according to the standard method. We prepared the extracts of plant with different solvents for determining the different extractive values by maceration, Soxhlet extraction, successive extraction process, and determination of ash values, pH value, moisture content, and phytochemical screening to show the presence of carbohydrates, phenolic compounds, flavonoids, alkaloids, proteins, saponins, and lipids in the drug extract and fluorescence analysis in different solvent. Analysis of pesticide residues, aflatoxin, and heavy metals are also performed

Protective Effect of Procyanidin B2 on Acute Liver Injury Induced by Aflatoxin B in Rats / 生物医学与环境科学（英文）

Objective@#This study aimed to explore the protective effect of procyanidin B2 (PCB2) on acute liver injury induced by aflatoxin B (AFB ) in rats.@*Methods@#Forty Sprague Dawley rats were randomly divided into control, AFB , AFB + PCB2, and PCB2 groups. The latter two groups were administrated PCB2 intragastrically (30 mg/kg body weight) for 7 d, whereas the control and AFB groups were given the same dose of double distilled water intragastrically. On the sixth day of treatment, the AFB and AFB + PCB2 groups were intraperitoneally injected with AFB (2 mg/kg). The control and PCB2 groups were intraperitoneally administered the same dose of dimethyl sulfoxide (DMSO). On the eighth day, all rats were euthanized: serum and liver tissue were isolated for further examination. Hepatic histological features were assessed by hematoxylin and eosin-stained sections. Weight, organ coefficient (liver, spleen, and kidney), liver function (serum alanine aminotransferase, aspartate aminotransferase, alkaline phosphatase, total bilirubin, and direct bilirubin), oxidative index (catalase, glutathione, superoxide dismutase, malondialdehyde, and 8-hydroxy-2'-deoxyguanosine), inflammation factor [hepatic interleukin-6 (IL-6) mRNA expression and serum IL-6], and bcl-2/bax ratio were measured.@*Results@#AFB significantly caused hepatic histopathological damage, abnormal liver function, oxidative stress, inflammation, and bcl-2/bax ratio reduction compared with DMSO-treated controls. Our results indicate that PCB2 treatment can partially reverse the adverse liver conditions induced by AFB .@*Conclusion@#Our findings indicate that PCB2 exhibits a protective effect on acute liver injury induced by AFB .

Research progress on growth of Aspergillus flavus and its toxins production controlled by plant essential oils and its active ingredients / 中草药

Aspergillus flavus is a pathogenic fungus, which can easily contaminate Chinese herbal medicines, food, and agricultural commodities. Aflatoxin (AFT) produced by the secondary metabolism of A. flavus caused serious threats to the health of people and animals due to its carcinogenic and highly toxic characteristics. Therefore, the search for inhibitors to A. flavus and its toxins has been widely concerned in the world. Essential oil is a natural antifungal agent extracted and highly concentrated from higher plants. It has the advantages of good antifungal effect, low pollution, and relatively safe to human body. Some kinds of them, including Thymus vulgaris, Origanum virens, Perilla frutescens, Foeniculum vulgare, and so on, displayed their antifungal activities. Specifically, active ingredients such as alcohols, aldehydes, and phenols have relatively good antifungal effects. The research progress on pathogenicity of A. flavus and AFT and the prevention and control by essential oils are reviewed in this article, which provided a reference for further exploring the antifungal mechanism of essential oils and developing natural and green antifungal products.

Preparation of highly sensitive monoclonal antibody against aflatoxin B_1 and its application in rapid detection of contamination in Ziziphi Spinosae Semen / 中国中药杂志

A highly sensitive monoclonal antibody against aflatoxin B_1(AFB_1) was prepared and an indirect competition enzyme-linked immunosorbent assay(ic-ELISA) was established based on the antibody which was used for high-throughput and rapid screening of AFB_1 contamination in Chinese herbal medicines to ensure the safety of medication. In this study, the structure of AFB_1 was modified by improved oxime method, and the carrier protein was coupled by EDC-NHS method to obtain the complete antigen of AFB_1, which was more convenient and environmental friendly. The Balb/c female mice were immunized using increasing the immunization dose and various ways of injection, and finally the AFB_1 monoclonal antibody was prepared. The AFB_1 monoclonal antibody belongs to IgG_(2 b) immunoglobulin by identifying its immunological characteristics, and its sensitivity(IC_(50)) can reach 0.15 μg·L~(-1), and the affi-nity is 2.81×10~8 L·mol~(-1). The cross-reaction rates of AFB_2, AFG_1, and AFG_2 were 35.07%, 8.75%, and 1.15%, respectively, and there was almost no cross-reactivity with other mycotoxins. Based on the high sensitivity and specificity of the antibody, an ic-ELISA method was established and applied to the determination of AFB_1 contamination in Ziziphi Spinosae Semen. According to the matrix matching standard curve, the linear concentration range for AFB_1 was 0.05-0.58 μg·L~(-1)(R~2=0.992), the recoveries were 88.00%-119.0%, and the detection limit was 1.69 μg·kg~(-1). The AFB_1 in 33 batches of Ziziphi Spinosae Semen samples was determined by ic-ELISA, and the contamination level was 3.62-206.58 μg·kg~(-1). The linear correlation coefficient between the detection results of ic-ELISA and UHPLC-MS/MS was 0.996, and there were no false positive and false negative cases. It indicates that the established ic-ELISA is accurate and reliable, and could provide a simple and effective technique for fast screening of AFB_1 contamination in Ziziphi Spinosae Semen, and also could be considered as the reference for the detection and monitoring of AFB_1 contamination in other Chinese herbal medicines.