Mohamed Amine Gacem，Aminata Ould El Hadj-Khelil

1Laboratory of Protection of Ecosystems in Arid and Semi-Arid Area，University of Kasdi Merbah，Ouargla 30000，Algeria

2Department of Biology，Faculty of Science，University of Amar Tlidji，Laghouat 03000，Algeria

Toxicology，biosynthesis，bio-control of aflatoxin and new methods of detection

Mohamed Amine Gacem1，2*，Aminata Ould El Hadj-Khelil1

1Laboratory of Protection of Ecosystems in Arid and Semi-Arid Area，University of Kasdi Merbah，Ouargla 30000，Algeria

2Department of Biology，Faculty of Science，University of Amar Tlidji，Laghouat 03000，Algeria

ARTICLE INFO

Article history:

Accepted 14 Mar 2016

Available online 4 Aug 2016

Mycotoxins

Af?latoxins

Cancer

Biosynthesis

Bio-controls

Detection methods

Mycotoxins and their derivatives since their discoveries and until the present time are behind unspecified economic and medical damages.Aflatoxins are classified according to their physical-chemical and toxicological characters in the most dangerous row of the mycotoxins.These aflatoxins are in part responsible，of irreversible medical disasters that are not easily manageable such as cancer of the liver and kidneys，and in the other part，of losses in the stored cereal products.Based on these crucial findings，monitoring of this toxin became imperative in post-harvest food products，during storage，during transformation chain and even during the long phases of conservation.Vigilance of this toxin is delivered by detection methods using very advanced technologies to respond in the shortest possible times.In addition，the knowledge of factors supporting the biosynthesis of aflatoxins such as the temperature，moisture content，concentration of nitrogen and carbon，and the molecules responsible for the genetic control of the synthesis will be reflected later in the choice of bio-control techniques.This control is currently based on new strategies using the bioactives substances of the plants，the lactic bacteria and some strains of actinomycetes that have good inhibiting activity against aflatoxins with fewer side effects on Man.On the other hand，this brief review summarizes the results of new studies demonstrating the toxicity of the toxin，new detection methods and bio-control.

1.Introduction

Mycotoxins are a much diversified group of toxic compounds produced by five kinds of spore-forming fungi，known to cause noxious effects to the health of human and animals.Food security is regularly risked by mycotoxins appearing in food［1，2］.

Amongst the mycotoxins，aflatoxins are most intensively sought because of their immunotoxicity acting on phagocytes and cell-mediated immunity［3］.They are regarded as natural contaminants from a large variety of agricultural products such as maize［4］.These compounds also affect a wide range of foods and fermented foods because of their richness in nutrients promoting their syntheses［5，6］.Their threshold can exceed the standards set by the European Union as in dry sweet chestnut occasionally consumed［7，8］.

The producing fungi of these types of mycotoxins can develop inside some food when the environmental conditions are favorable to the biosynthesis［9］.Slightly higher CO2concentrations，interactions with the temperature and the availability of water can stimulate the growth of some mycotoxigenic species，especially under hydrous stress［10］.

Aflatoxins are mainly produced by the moulds belonging to the species Aspergillus［11-14］such as Aspergillus flavus（A.flavus），Aspergillus nomius，and Aspergillus parasiticus（A.parasiticus）［15］.Aflatoxins B1，B2，G1，and G2are most significant contaminants to rice［16］.Two other metabolites: aflatoxins M1and M2can be separated from milk［17，18］.

These poisons cause very dangerous effects in the consumer: these include carcinogenic，mutagen and teratogenic effects［19］. Consumption of aflatoxins contaminated corn has been found associated with an increased risk of cancer of the liver and acute hepatitis in certain areas of the South Africa and China［20，21］.A study showed that the consumption of groundnuts contaminated by aflatoxins caused the death of several cows with hepatic damage according to the histological analysis［22］. Strong exposure to aflatoxins causes growth delay in young children［23］.

Aflatoxin B1（AFB1）is the most relevant mycotoxin because of its toxic effect in humans［13］，it is found with higher concentrations in contaminated food［24］.AFB1is particularly toxic because of its role in liver cancer［17，18］.

The objective of this literature study is to collect maximum information and clarifications argued with recent references to present a bibliographical review covering the toxicological profile of AFB1，the new methods of bio-control and recent techniques available for detecting this toxin.

2.Recent studies showing the toxicity of AFB1

AFB1is a powerful carcinogen harmful to health as it causes lung and liver cancer［25，26］.The International Agency for Research on Cancer recognized that AFB1and aflatoxin M1are carcinogenic from Group 1 for human and animals［27］.

Recent studies carried out on the mice showed that the early exposure to the AFB1in particular at the embryonic period is a mutagen［28］，as it causes a reduction of the body weight，a reduction in the weight of the reproductive organs，a reduction in the number and mobility of the spermatozoa with a lowering of the rate of serum testosterone and the enzymes of the steroidogenesis［29］.

The cytotoxicity was shown in vivo on the renal cells of a monkey.The results showed that this toxin causes a considerable reduction in the viable cells（37%），it also causes oxidative damage by enhancing the peroxidation of lipids［30］.In Brazil，aflatoxin M1was detected in the human urines with a going rate from 0.19 to 12.7 pg/mg，whereas no residues of aflatoxins B1，B2，G1and G2were identified［31］.The AFB1is toxic for human lymphocytes mediated by apoptosis and necrosis［32］.

In broiler chicken aflatoxins affect the pancreatic activity，resulting in histological changes of the organ，with an increase in the activity of lipase and alpha amylase，and the activity of trypsin is also affected［33］.

3.Major factors influencing biosynthesis of AFB1

The biosynthesis of the AFB1requires several steps（Figure 1）and it is perhaps affected by the intervention of several environmental factors（stress，quorum sensing and protein signaling pathway）without forgetting the factors regulating the transcription unit［34］.

Amino-acids such as tryptophan inhibit the synthesis of aflatoxin whereas tyrosin encourages it［35］.The presence of the lipids induces the aflatoxinogenesis［36］.Among the organic factors affecting biosynthesis，carbon and nitrogen are the major ones［37］.In addition，simple sugars such as glucose and fructose support this biosynthesis，whereas in the cases of sorbose and lactose no action has been recorded［38］.

Concerning the physical factors，the optimal temperature of biosynthesis is located between 28°C and 35°C.Above this temperature range，biosynthesis is inhibited due to the attack of transcription genes aflR and aflS［39，40］，whereas under the conditions of dryness，the production of the aflatoxins is high［41］.Synthesis is also influenced by subcultures and changes in the morphology of producing cells［42］.For pH，biosynthesis is high in acidic mediums while it is inhibited in basic conditions［43］，for A.parasiticus，the growth in water is faster with a pH ranging from 5.5 to 6.5［44］.

The secondary plant metabolites play a key role in the synthesis of aflatoxins［45］.For example，the presence of the octanal causes a reduction of 60%of the fungic growth with a rate of increase in the production of aflatoxins of 500%［46］. However，hydrolysable tannins considerably inhibit the biosynthesis of aflatoxins［47］.Some antioxidants such as the phenolic compounds，ascorbic acid and caffeic acid decrease，in an important way，the aflatoxinogenesis，without any effect on the growth of the fungi［48，49］.

4.Detoxification and bio-control of aflatoxins

Harmful effects caused by this dangerous toxin have directed researchers towards finding new strategies for prevention and detoxification in order to preserve the safety of products intended for human consumption［50］.

4.1.Detoxification using probiotics and lactic acid bacteria

Several lactic bacteria are able to bind AFB1in vitro and in vivo on the surface of the organism and take two aspects were into consideration:binding and release of toxin［51］.Turbic and his collaborators showed that 77%-95%of AFB1were removed by strains of Lactobacillus rhamnosus GG and LC-705［52］.El Khoury and his collaborators also noted that Lactobacillus bulgaricus and Streptococcus thermophilus were effective in the reduction of aflatoxins M1［53］.Lactobacillus pentosus and Lactobacillus beveris have the capacity to absorb and release AFB1［54］.

The activity of Lactobacillus plantarum（L.plantarum）is studied in Tunisia on olives（Chetoui varieties）collected in 2008-2009 and 2009-2010.Samples were prepared and then inoculated by A.flavus and L.plantarum.Results of crude samples showed that samples of 2008-2009 were salubrious due to the presence of antimicrobial substances and the absence of the biosynthesis of this toxin［55］.The identification of mycoflore revealed presence of species belonging to genus Candida，Rhodotorula，Crytococcus，Pichia，Aspergillus，Geotrichum，Penicillium with absence of AFB1producing species.For thoseof2009-2010andatarateof2.106cell/gof L.plantarum AFB1was reduced from 11.0μg/kg up to 5.9μg/kg［55］.

L.plantarum adheres to the surface of olive and produces biofilms，which affects adherence of other undesirable microorganisms，supports the increase in antioxidant activity and consequently，it weakens the production of AFB1［55］.This reduction is also due to attraction of oxygen by L.plantarum thus protecting of polyols against oxidation and increasing inhibition of AFB1biosynthesis［55］，According to other studies，such reduction of AFB1rate is due to the binding of toxin by reversible physical bonds［56］，binding with certain molecules in the wall of the L.rhamnosus GG［57］，or by synthesis of extracellular polysaccharides trapper of radicals and having an antioxidant activity in some probiotics like Bacillus coagulans RK-02［58］.Other strains of Bacillus spp. have the ability to degrade AFB1［59］.

According to Magnusson and his collaborators，three mechanisms can explain the antimicrobial effectiveness of the lactic bacteria，namely，organic acid production，competition for nutritive element and antagonist production［60］.

Figure 1.Biosynthesis steps of aflatoxin.

4.2.Detoxification using bioactive substances of plants

Phenolic pulp extract of Dialium guineense has been proved effective for sweeping and trapping of oxygenated chemical species and the prevention of lipid peroxidation，proteins oxidation and DNA fragmentation by AFB1［61］.Another adsorbent prepared from bagasse has been proved effective for detoxification of AFB1in the gastrointestinal tract of chicks and no negative symptoms was reported［62］.

Aflatoxin oxidase，an enzyme of Armillariella tabescens presents a detoxifying activity towards AFB1.This reaction is dependant on oxygen and hydrogen peroxide producing，which may play the crucial role in detoxification of aflatoxin oxidase［63］.Laccase is another enzyme that has proven its detoxifying affinity for AFB1［64］.Furthermore，manganese peroxidase is an enzyme of Pleurotus ostreatus which may detoxify AFB1according to the enzyme concentration and the incubation period，this detoxification can reach 90%at 1.5 IU/mL of enzyme during 48 h of incubation［65］.The treatment with kaempferol also decreases toxic effects of AFB1［66］.

4.3.Detoxification using biomolecules of fungi

Fungal proteins are molecules of small sizes very basic and rich in cysteine such as PgAFP，NFAP and PC-Arctin［67-69］. The NFAP induces an oxidative stress in sensitive fungi causing the apoptosis［70］，whereas，PgAFP inhibits growth in some toxigenic molds［67］.A recent study proved a reduction growth of A.flavus with significant changes including several proteins at concentration higher than 9.38μg/mL of PgAFP. Cells treated by PgAFP showed a more intense oxidation［71］.

4.4.Detoxification using actinomycetes

New control strategies have been developed in recent years based on the use of actinomycetes.A study conducted on Cuban soil led to isolation of 563 actinomycetes，in which 50.7% have an antifungal activity against Candida，Trichophyton mentagrophytes，Penicillium chrysogenum and Colletotrichum musae，probably due to the production of antibiotics belonging to the families of aminoglycoside，anthracycline or polyether ionophore and polyene macrolide antibiotic［72］.Another study conducted by Okudoh and Wallis in 2007 revealed that actinomycetes isolates from forests soil had an antimicrobial activity better than those from the riparian soil.Isolates from poultry manure，straws，chickens and composts soil had inhibition zones varying between 20 and 30 mm against Candida utilis［73］.

In 2013，a study showed that Streptomyces phaeochromogenes isolated from soil of a garden of Sagar，Madhya Pradesh showed a very good inhibiting activity against Candida albicans due to an intracellular antibiotic with a 27 mm inhibition zone［74］.In addition，a new strain identified and named Streptomycessp.PP14 has just been isolated from the Canadian soil after their thermal and chemical treatment with phenol in order to have resistant strains.This strain showed a very good activity against Penicillium expansum，Fusarium graminearum，A. flavus and Aspergillus carbonarius with inhibition zones varying between 17 and 45 mm on solid medium ISP2［75］. Recently，a study allowed to isolate 37 strains of actinomycetes from various areas of Algeria.These strains showed a very good activity against A.flavus NRRL 62477. Their culture in a medium containing 5 mg/kg of pure AFB1allowed to have a reduced rate which can go up to（15.6±11.7）AFB1（residual concentration in the medium in %）［76］.

4.5.Detoxification using other methods

A study was performed on 192 laying hens in order to evaluate the effect of vitamin E on AFB1administration.The results revealed that eggs production was reduced completely after three weeks of exposure to AFB1（10μg/kg）and vitamin E（0.1 mg/kg），the shell thickness was less than normal，AFB1was present in eggs at 2.5μg/kg despite of the presence of vitamin E，the latter recorded an antioxidant effect against hemolyses caused by 2，2ʹ-azobis（2-amidino-propane）hydrochloride［77］.

Another study was carried out on six groups of Swiss albino mice in order to determine the protective effect of esculin and ascorbic acid against AFB1，and it showed that AFB1causes 387%of lipid peroxidation of necrosis and degeneration in renal tubules.The addition of esculin or the ascorbic acid decreased this effect，which is due to on the one hand the controlling of reduced glutathione，glutathione peroxidase，glutathione-Sreductase，glutathione reductase，superoxide dismutase and catalase and on the other hand，the regeneration of the renal tubules and the glomerular epithelial cells［78］.

5.New methods of detection of the AFB1

Detection methods of AFB1has underwent remarkable development since its discovery.Thin layer chromatography is one of the oldest techniques used to analyze contaminated samples［79］.Other methods are also used such as high performance liquid chromatography with fluorescent detector［80］，or with fluorimetric detector［81］.Aflatoxins are also detected by liquid chromatography coupled to a mass spectrometer［82］.Other methods of detection were elaborated such as immune-affinity column immune-enzymatic and immunochemical methods［83，84］.

Detection of ultra-traces of AFB1is extremely important for food safety，this detection requires very powerful techniques. Among the preview of these techniques during this bibliographic research，an aptasensor using unmodified gold nanoparticules indicator based on the aggregation phenomenon of gold nanoparticules induced by salt was recently developed［85］.Another very recent technique by electrochemical immunosensor sensitive to AFB1based on carbon nanotubes with simple walled，this immunosensor was based on an indirect competitive binding.The detection limit is 3.5 pg/mL.In addition，the immunosensor was successfully applied for determination of AFB1in corn powder，which showed a good correlation with the results obtained by high performance liquid chromatography［86］.

Technique ultra-high pressure liquid chromatography tandem mass spectrometry was developed to identify and quantify simultaneously mycotoxins in ensiling grasses.It was performed using a modified QuEChERS extraction by employing an acidified aqueous extraction［87］.Another uncomplicated technique with a detection limit of 0.03 ng/mL for AFB1based on sensitive surface-enhanced Raman scattering beacons has been developed without nucleic acid amplification［88］.

Other ultra-sensitive strategies；colorimetric and homogeneous for AFB1detection were set up using a DNA aptamer，and two halves of split DNAzyme has been developed［89］.

Another inexpensive method，free from any interference and even automated has been developed successfully.This technique is used for the detection of AFM1 in milk based upon microextraction in the liquid phase of hollow fiber coupled with the liquid chromatography/tandem mass spectrometry［90］.Visual detection method has undergone a remarkable development.In this review，a visual detection method was implemented using chemical reactions with dichlorvos-ammonia on agar cultures，wherein dichlorvos inhibits esterase causing accumulation of anthraquinone precursors of aflatoxins mycelium to agar，followed by a change in the color of colonies from light yellow to purple red by treatment with ammonia vapor［91］.

In order to facilitate the bio-monitoring which provides the best approach to evaluate the human exposure to the mycotoxins，the detection of AFM1 in urines（derived from the decomposition of AFB1）was performed by the immuneenzymatic method［92］.

6.Conclusions

In this literature review，all information collected were obtained from very recent studies.The information presented in this paper has demonstrated the toxic effects of AFB1.Biomonitoring and bio-control are two crucial points in order to limit all undesirable effects or to make them reversible.

Conflict of interest statement

We declare that we have no conflict of interest.

Acknowledgments

We are thankful to the researchers from the Laboratory of Protection of Ecosystems in Arid and Semi-Arid Area，University of Kasdi Merbah-Ouargla，Algeria for their support and for providing valuable information during this work.

References

［1］D'Mello JPF.Food safety:contaminants and toxins.Edinburgh: CABI；2003，p.65-6.

［2］Ilesanmi FF，Ilesanmi OS.Knowledge of aflatoxin contamination in groundnut and the risk of its ingestion among health workers in Ibadan，Nigeria.Asian Pac J Trop Biomed 2011；1（6）:493-5.

［3］Bondy GS，Pestka JJ.Immunomodulation by fungal toxins. J Toxicol Environ Health B Crit Rev 2000；3（2）:109-43.

［4］Mutiga SK，Hoffmann V，Harvey JW，Milgroom MG，Nelson RJ. Assessment of aflatoxin and fumonisin contamination of maize in Western Kenya.Phytopathology 2015；105（9）:1250-61.

［5］Yang ZY，Shim WB，Kim JH，Park SJ，Kang SJ，Nam BS，et al. Detection of aflatoxin-producing molds in Korean fermented foods and grains by multiplex PCR.J Food Prot 2004；67（11）:2622-6.

［6］ElKhouryA，RizkT，LteifR，AzouriH，DeliaML，LebrihiA.Fungal contamination and aflatoxin B1and ochratoxin A in Lebanese winegrapes and musts.Food Chem Toxicol 2008；46（6）:2244-50.

［7］Pietri A，Rastelli S，Mulazzi A，Bertuzzi T.Aflatoxins and ochratoxin A in dried chestnuts and chestnut flour produced in Italy. Food Control 2012；25（2）:601-6.

［8］Bertuzzi T，Gualla A，Morlacchini M，Pietri A.Direct and indirect contamination with ochratoxin A of ripened pork products.Food Control 2013；34（1）:79-83.

［9］Georgiadou M，Dimou A，Yanniotis S.Aflatoxin contamination in pistachio nuts:a farm to storage study.Food Control 2012；26（2）: 580-6.

［10］Magan N，Medina A，Aldred D.Possible climate-change effects on mycotoxin contamination of food pre-and post-harvest.Plant Pathol 2011；60（1）:150-63.

［11］Mayer Z，Bagnara A，F¨arber P，Geisen R.Quantification of the copy number of nor-1，a gene of the aflatoxin biosynthetic pathway by real-time PCR，and its correlation to the cfu of Aspergillus flavus in foods.Int J Food Microbiol 2003；82（2）:143-51.

［12］Schmidt-Heydt M，Abdel-Hadi A，Magan N，Geisen R.Complex regulation of the aflatoxin biosynthesis gene cluster of Aspergillus flavus in relation to various combinations of water activity and temperature.Int J Food Microbiol 2009；135（3）:231-7.

［13］Rodríguez A，Rodríguez M，Martín A，Nuñez F，C´ordoba JJ. Evaluation of hazard of aflatoxin B1，ochratoxin A and patulin production in dry-cured ham and early detection of producing moulds by qPCR.Food Control 2012；27（1）:118-26.

［14］Corcuera LA，Vettorazzi A，Arbillaga L，Gonz´alez-Peñas E，de Cerain AL.An approach to the toxicity and toxicokinetics of aflatoxin B1and ochratoxin A after simultaneous oral administration to fasted F344 rats.Food Chem Toxicol 2012；50（10）:3440-6.

［15］Wilson DM，Mubatanhema W，Jurjevic Z.Biology and ecology of mycotoxigenic Aspergillus species as related to economic and health concerns.Adv Exp Med Biol 2002；504:3-17.

［16］Su´arez-Bonnet E，CarvajalM，M´endez-Ramírez I，Castillo-Urueta P，Cort´es-Eslava J，G´omez-Arroyo S，et al.Aflatoxin（B1，B2，G1，and G2）contamination in rice of Mexico and Spain，from local sources or imported.J Food Sci 2013；78（11）:T1822-9.

［17］Pitt JI.Toxigenic fungi and mycotoxins.Br Med Bull 2000；56（1）: 184-92.

［18］Alberts JF，Engelbrecht Y，Steyn PS，Holzapfel WH，van Zyl WH. Biological degradation of aflatoxin B1by Rhodococcus erythropolis cultures.Int J Food Microbiol 2006；109（1-2）:121-6.

［19］Krska R，Schubert-Ullrich P，Molinelli A，Sulyok M，MacDonald S，Crews C.Mycotoxin analysis:an update.Food Addit Contam Part A 2008；25（2）:152-63.

［20］Wogan GN.Aflatoxin as a human carcinogen.Hepatology 1999；30（2）:573-5.

［21］Li FQ，Yoshizawa T，Kawamura O，Luo XY，Li YW.Aflatoxins and fumonisins in corn from the high-incidence area for human hepatocellular carcinoma in Guangxi，China.J Agric Food Chem 2001；49（8）:4122-6.

［22］RayAC，AbbittB，CotterSR，MurphyMJ，ReagorJC，Robinson RM，et al.Bovine abortion and death associated with consumption of aflatoxin-contaminated peanuts.J Am Vet Med Assoc 1986；188（10）:1187-8.

［23］Lombard MJ.Mycotoxin exposure and infant and young child growth in Africa:what do we know？Ann Nutr Metab 2014；64（Suppl 2）:42-52.

［24］Park JW，Kim EK，Shon DH，Kim YB.Natural co-occurrence of aflatoxin B1，fumonisin B1and ochratoxin A in barley and corn foods from Korea.Food Addit Contam 2002；19（11）:1073-80.

［25］Dvorackova I，Stora C，Ayraud N.Evidence of aflatoxin B1in two cases of lung cancer in man.J Cancer Res Clin Oncol 1981；100（2）:221-4.

［26］Kelly JD，Eaton DL，Guengerich FP，Coulombe RA Jr.Aflatoxin B1activation in human lung.Toxicol Appl Pharmacol 1997；144（1）:88-95.

［27］Fern´andez-Ibañez V，Soldado A，Martínez-Fern´andez A，de la Roza-Delgado B.Application of near infrared spectroscopy for rapid detection of AFB1in maize and barley as analytical quality assessment.Food Chem 2009；113（2）:629-34.

［28］Chawanthayatham S，Thiantanawat A，Egner PA，Groopman JD，Wogan GN，Croy RG，et al.Prenatal exposure of mice to the human liver carcinogen aflatoxin B1reveals a critical window of susceptibility to genetic change.Int J Cancer 2015；136（6）:1254-62.

［29］Supriya C，Girish BP，Reddy PS.Aflatoxin B1-induced reproductive toxicity in male rats:possible mechanism of action.Int J Toxicol 2014；33（3）:155-61.

［30］V´azquez-Dur´an A，Díaz-Torres R，Ramírez-Noguera P，Moreno-Martínez E，M´endez-Albores A.Cytotoxic and genotoxic evaluation of tortillas produced by microwave heating during alkalinecooking of aflatoxin-contaminated maize.J Food Sci 2014；79（5）:T1024-9.

［31］Jager AV，Tonin FG，Souto PC，Privatti RT，Oliveira CA.Determination of urinary biomarkers for assessment of short-term human exposure to aflatoxins in São Paulo，Brazil.Toxins（Basel）2014；6（7）:1996-2007.

［32］Al-Hammadi S，Marzouqi F，Al-Mansouri A，Shahin A，Al-Shamsi M，Mensah-Brown E，et al.The cytotoxicity of aflatoxin B1in human lymphocytes.Sultan Qaboos Univ Med J 2014；14（1）: e65-71.

［33］Marchioro A，Mallmann AO，Diel A，Dilkin P，Rauber RH，Blazquez FJ，et al.Effects of aflatoxins on performance and exocrine pancreas of broiler chickens.Avian Dis 2013；57（2）:280-4.

［34］Amare MG，Keller NP.Molecular mechanisms of Aspergillus flavus secondary metabolism and development.Fungal Genet Biol 2014；66:11-8.

［35］Wilkinson JR，Yu J，Bland JM，Nierman WC，Bhatnagar D，Cleveland TE.Amino acid supplementation reveals differential regulation of aflatoxin biosynthesis in Aspergillus flavus NRRL 3357 and Aspergillus parasiticus SRRC 143.Appl Microbiol Biotechnol 2007；74（6）:1308-19.

［36］Yu J，Mohawed SM，Bhatnagar D，Cleveland TE.Substrateinduced lipase gene expression and aflatoxin production in Aspergillus parasiticus and Aspergillus flavus.J Appl Microbiol 2003；95（6）:1334-42.

［37］Luchese RH，Harrigan W F.Biosynthesis of aflatoxin-the role of nutritional factors.J Appl Bacteriol 1993；74（1）:5-14.

［38］Payne GA，Brown MP.Genetics and physiology of aflatoxin biosynthesis.Annu Rev Phytopathol 1998；36:329-62.

［39］O'Brian GR，Georgianna DR，Wilkinson JR，Yu J，Abbas HK，Bhatnagar D，et al.The effect of elevated temperature on gene transcription and aflatoxin biosynthesis.Mycologia 2007；99（2）: 232-9.

［40］Yu J，Fedorova ND，Montalbano BG，Bhatnagar D，Cleveland TE，Bennett JW，et al.Tight control of mycotoxin biosynthesis gene expression in Aspergillus flavus by temperature as revealed by RNA-Seq.FEMS Microbiol Lett 2011；322（2）:145-9.

［41］Cotty PJ，Jaime-Garcia R.Influences of climate on aflatoxin producing fungi and aflatoxin contamination.Int J Food Microbiol 2007；119（1-2）:109-15.

［42］Torres J，Guarro J，Suarez G，Suñe N，Calvo MA，Ramírez C. Morphological changes in strains of Aspergillus flavus Link ex Fries and Aspergillus parasiticus Speare related with aflatoxin production.Mycopathologia 1980；72（3）:171-80.

［43］Cotty PJ.Aflatoxin and sclerotial production by Aspergillus flavus: influence of pH.Phytopathology 1988；78（09）:1250-3.

［44］Al-Gabr HM，Ye C，Zhang Y，Khan S，Lin H，Zheng T.Effects of carbon，nitrogen and pH on the growth of Aspergillus parasiticus and aflatoxins production in water.J Environ Biol 2013；34（2 Spec No）:353-8.

［45］Greene-McDowelle DM，Ingber B，Wright MS，Zeringue HJ Jr，Bhatnagar D，Cleveland TE.The effects of selected cotton-leaf volatiles on growth，development and aflatoxin production of Aspergillus parasiticus.Toxicon 1999；37（6）:883-93.

［46］Criseo G，Bagnara A，Bisignano G.Differentiation of aflatoxinproducing and non-producing strains of Aspergillus flavus group. Lett Appl Microbiol 2001；33（4）:291-5.

［47］Mahoney N，Molyneux RJ.Phytochemical inhibition of aflatoxigenicity in Aspergillus flavus by constituents of walnut（Juglans regia）.J Agric Food Chem 2004；52（7）:1882-9.

［48］Bok JW，Keller NP.LaeA，a regulator of secondary metabolism in Aspergillus spp.Eukaryot Cell 2004；3（2）:527-35.

［49］Kim JH，Campbell BC，Molyneux R，Mahoney N，Chan KL，Yu J，et al.Gene targets for fungal and mycotoxin control.Mycotoxin Res 2006；22（1）:3-8.

［50］Ben Salah-Abb`es J，Abb`es S，Jebali R，Haous Z，Oueslati R.Potential preventive role of lactic acid bacteria against aflatoxin M1immunotoxicity and genotoxicity in mice.J Immunotoxicol 2015；12（2）:107-14.

［51］Bueno DJ，Casale CH，Pizzolitto RP，Salvano MA，Oliver G. Physical adsorption of aflatoxin B1by lactic acid bacteria and Saccharomyces cerevisiae:a theoretical model.J Food Prot 2007；70（9）:2148-54.

［52］Turbic A，Ahokas JT，Haskard CA.Selective in vitro binding of dietary mutagens，individually or in combination，by lactic acid bacteria.Food Addit Contam 2002；19（2）:144-52.

［53］El Khoury A，Atoui A，Yaghi J.Analysis of aflatoxin M1 in milk and yogurt and AFM1 reduction by lactic acid bacteria used in Lebanese industry.Food Control 2011；22（10）:1695-9.

［54］Hamidi A，Mirnejad R，Yahaghi E，Behnod V，Mirhosseini A，Amani S，et al.The aflatoxin B1isolating potential of two lactic acid bacteria.Asian Pac J Trop Biomed 2013；3（9）:732-6.

［55］Kachouri F，Ksontini H，Hamdi M.Removal of aflatoxin B1and inhibition of Aspergillus flavus growth by the use of Lactobacillus plantarum on olives.J Food Prot 2014；77（10）:1760-7.

［56］Haskard CA，El-Nezami HS，Kankaanp¨a¨a PE，Salminen S，Ahokas JT.Surface binding of aflatoxin B1by lactic acid bacteria. Appl Environ Microbiol 2001；67（7）:3086-91.

［57］LahtinenSJ，HaskardCA，OuwehandAC，SalminenSJ，Ahokas JT.Binding of aflatoxin B1to cell wall components of Lactobacillus rhamnosus strain GG.Food Addit Contam 2004；21（2）:158-64.

［58］Kodali VP，Sen R.Antioxidant and free radical scavenging activities of an exopolysaccharide from a probiotic bacterium.Biotechnol J 2008；3（2）:245-51.

［59］Galarza-SeeberR，LatorreJD，Hernandez-VelascoX，Wolfenden AD，Bielke LR，Menconi A，et al.Isolation，screening and identification of Bacillus spp.as direct-fed microbial candidates for aflatoxin B1biodegradation.Asian Pac J Trop Biomed 2015；5（9）:702-6.

［60］Magnusson J，Str¨om K，Roos S，Sj¨ogren J，Schn¨urer J.Broad and complex antifungal activity among environmental isolates of lactic acid bacteria.FEMS Microbiol Lett 2003；219（1）:129-35.

［61］Adeleye AO，Ajiboye TO，Iliasu GA，Abdussalam FA，Balogun A，Ojewuyi OB，et al.Phenolic extract of Dialium guineense pulp enhances reactive oxygen species detoxification in aflatoxin B1 hepatocarcinogenesis.J Med Food 2014；17（8）:875-85.

［62］Khan FA，Zahoor M.In vivo detoxification of aflatoxin B1 by magnetic carbon nanostructures prepared from bagasse.BMC Vet Res 2014；10:255.

［63］Wu YZ，Lu FP，Jiang HL，Tan CP，Yao DS，Xie CF，et al.The furofuran-ring selectivity，hydrogen peroxide-production and low Km value are the three elements for highly effective detoxification of aflatoxin oxidase.Food Chem Toxicol 2015；76:125-31.

［64］Zeinvand-Lorestani H，Sabzevari O，Setayesh N，Amini M，Nili-Ahmadabadi A，Faramarzi MA.Comparative study of in vitro prooxidative properties and genotoxicity induced by aflatoxin B1and its laccase-mediated detoxification products.Chemosphere 2015；135:1-6.

［65］Yehia RS.Aflatoxin detoxification by manganese peroxidase purified from Pleurotuso streatus.Braz J Microbiol 2014；45（1）:127-33.

［66］Langeswaran K，Revathy R，Kumar SG，Vijayaprakash S，Balasubramanian MP.Kaempferol ameliorates aflatoxin B1（AFB1）induced hepatocellular carcinoma through modifying metabolizing enzymes，membrane bound ATPases and mitochondrial TCA cycle enzymes.Asian Pac J Trop Biomed 2012；2（3）:S1653-9.

［67］Rodríguez-Martín A，Acosta R，Liddell S，N´uñez F，Benito MJ，Asensio MA.Characterization of the novel antifungal protein PgAFP and the encoding gene of Penicillium chrysogenum.Peptides 2010；31（4）:541-7.

［68］Kov´acs L，Vir´agh M，Tak´o M，Papp T，V´agv¨olgyi C，Galg´oczy L. Isolation and characterization of Neosartorya fischeri antifungal protein（NFAP）.Peptides 2011；32（8）:1724-31.

［69］Chen Z，Ao J，Yang W，Jiao L，Zheng T，Chen X.Purification and characterization of a novel antifungal protein secreted by Penicillium chrysogenum from an Arctic sediment.Appl Microbiol Biotechnol 2013；97（24）:10381-90.

［70］Galg´oczy L，Kov´acs L，Kar´acsony Z，Vir´agh M，Hamari Z，V´agv¨olgyi C.Investigation of the antimicrobial effect of Neosartorya fischeri antifungal protein（NFAP）after heterologous expression in Aspergillus nidulans.Microbiology 2013；159（Pt 2）: 411-9.

［71］Delgado J，Owens RA，Doyle S，Asensio MA，N´uñez F.Impact of the antifungal protein PgAFP from Penicillium chrysogenum on the protein profile in Aspergillus flavus.Appl Microbiol Biotechnol 2015；99（20）:8701-15.

［72］Iznaga Y，Lemus M，Gonz´alez L，Garmendía L，Nadal L，Vallín C. Antifungal activity of actinomycetes from Cuban soils.Phytother Res 2004；18（6）:494-6.

［73］Okudoh VI，Wallis FM.Antimicrobial activity of rare actinomycetes isolated from natural habitats in KwaZulu-natal，South Africa. S Afr J Sci 2007；103（5-6）:216-22.

［74］Saxena A，Upadhyay R，Kumar D，Kango N.Isolation，antifungal activity and characterization of soil actinomycetes.J Sci Ind Res 2013；72（8）:491-7.

［75］Bouras N，Meklat A，Toumatia O，Mokrane S，Holtz MD，Strelkov SE，et al.Bioactive potential of a new strain of Streptomyces sp.PP14 isolated from Canadian soil.Afr J Microbiol Res 2013；7（25）:3199-208.

［76］Verheecke C，Liboz T，Darriet M，Sabaou N，Mathieu F.In vitro interaction of actinomycetes isolates with Aspergillus flavus: impact on aflatoxins B1and B2production.Lett Appl Microbiol 2014；58（6）:597-603.

［77］KhanWA，KhanMZ，KhanA，HassanZU，RafiqueS，Saleemi MK，et al.Dietary vitamin E in White Leghorn layer breeder hens:a strategy to combat aflatoxin B1-induced damage. Avian Pathol 2014；43（5）:389-95.

［78］Naaz F，Abdin MZ，Javed S.Protective effect of esculin against prooxidant aflatoxin B1-induced nephrotoxicity in mice.Mycotoxin Res 2014；30（1）:25-32.

［79］Fallah AA，Rahnama M，Jafari T，Saei-Dehkordi SS.Seasonal variation of aflatoxin M1contamination in industrial and traditional Iranian dairy products.Food Control 2011；22（10）:1653-6.

［80］Andrade PD，da Silva JLG，Caldas ED.Simultaneous analysis of aflatoxins B1，B2，G1，G2，M1and ochratoxin A in breast milk by high-performance liquid chromatography/fluorescence after liquidliquid extraction with low temperature purification（LLE-LTP）. J Chromatogr A 2013；1304:61-8.

［81］Tabari M，Karim G，Ghavami M，Chamani M.Method validation for aflatoxin M1determination in yoghurt using immunoaffinity column clean-up prior to high-performance liquid chromatography. Toxicol Ind Health 2011；27（7）:629-35.

［82］Sulyok M，Beed F，Boni S，Abass A，Mukunzi A，Krska R. Quantitation of multiple mycotoxins and cyanogenic glucosides in cassava samples from Tanzania and Rwanda by an LC-MS/MS-based multi-toxin method.Food Addit Contam Part A Chem Anal Control Expo Risk Assess 2015；32（4）:488-502.

［83］Tang X，Li X，Li P，Zhang Q，Li R，Zhang W，et al.Development and application of an immunoaffinity column enzyme immunoassay for mycotoxin zearalenone in complicated samples.PLoS One 2014；9（1）:e85606.

［84］HeT，WangY，LiP，ZhangQ，LeiJ，ZhangZ，etal.Nanobody-based enzyme immunoassay for aflatoxin in agro-products with high tolerance to cosolvent methanol.Anal Chem 2014；86（17）:8873-80.

［85］Luan Y，Chen Z，Xie G，Chen J，Lu A，Li C，et al.Rapid visual detection of aflatoxin B1by label-free aptasensor using unmodified gold nanoparticles.J Nanosci Nanotechnol 2015；15（2）:1357-61.

［86］Zhang X，Li CR，Wang WC，Xue J，Huang YL，Yang XX，et al. A novel electrochemical immunosensor for highly sensitive detection of aflatoxin B1in corn using single-walled carbon nanotubes/chitosan.Food Chem 2016；192:197-202.

［87］McElhinney C，O'Kiely P，Elliott C，Danaher M.Development and validation of an UHPLC-MS/MS method for the determination of mycotoxins in grass silages.Food Addit Contam Part A Chem Anal Control Expo Risk Assess 2015；32（12）:2101-12.

［88］Zhao Y，Yang Y，Luo Y，Yang X，Li M，Song Q.Double detection of mycotoxins based on SERS labels embedded Ag@Au core-shell nanoparticles.ACS Appl Mater Interfaces 2015；7（39）:21780-6.

［89］Seok Y，Byun JY，Shim WB，Kim MG.A structure-switchable aptasensor for aflatoxin B1detection based on assembly of an aptamer/split DNAzyme.Anal Chim Acta 2015；886:182-7.

［90］Huang S，Hu D，Wang Y，Zhu F，Jiang R，Ouyang G.Automated hollow-fiber liquid-phase microextraction coupled with liquid chromatography/tandem mass spectrometry for the analysis of aflatoxin M1in milk.J Chromatogr A 2015；1416:137-40.

［91］Yabe K，Hatabayashi H，Ikehata A，Zheng Y，Kushiro M.Development of the dichlorvos-ammonia（DV-AM）method for the visual detection of aflatoxigenic fungi.Appl Microbiol Biotechnol 2015；99（24）:10681-94.

［92］Ali N，Hossain K，Blaszkewicz M，Rahman M，Mohanto NC，Alim A，et al.Occurrence of aflatoxin M1in urines from rural and urban adult cohorts in Bangladesh.Arch Toxicol 2015；90（7）: 1749-55.

16 Nov 2015

in revised form30Nov，2nd revised form31Dec2015，3rdrevised form 3 Feb 2016

Mohamed Amine Gacem，Department of Biology，Faculty of Science，University of Amar Tlidji，Laghouat 03000，Algeria.

Tel:+213 554 010 916

E-mail:biologieamine@yahoo.fr

Peer review under responsibility of Hainan Medical University.The journal implements double-blind peer review practiced by specially invited international editorial board members.

2221-1691/Copyright©2016 Hainan Medical University.Production and hosting by Elsevier B.V.This is an open access article under the CC BY-NC-ND license（http:// creative commons.org/licenses/by-nc-nd/4.0/）.