The occurrence and effect of unit operations for dairy products processing on the fate of aflatoxin M1: A review

Accepted Manuscript The occurrence and effect of unit operations for dairy products processing on the fate of aflatoxin M1: a review Fernanda Bovo Campagnollo, Karina C. Ganev, Amin Mousavi Khaneghah, Jéssica Portella, Adriano G. Cruz, Daniel Granato, Carlos H. Corassin, Carlos Augusto F. Oliveira, Anderson de Souza Sant’Ana PII:

S0956-7135(16)30176-1

DOI:

10.1016/j.foodcont.2016.04.007

Reference:

JFCO 4966

To appear in:

Food Control

Received Date: 22 February 2016 Revised Date:

5 April 2016

Accepted Date: 6 April 2016

Please cite this article as: Campagnollo F.B., Ganev K.C., Khaneghah A.M., Portella J., Cruz A.G., Granato D., Corassin C.H., Oliveira C.A.F. & de Souza Sant’Ana A., The occurrence and effect of unit operations for dairy products processing on the fate of aflatoxin M1: a review, Food Control (2016), doi: 10.1016/j.foodcont.2016.04.007. This is a PDF file of an unedited manuscript that has been accepted for publication. As a service to our customers we are providing this early version of the manuscript. The manuscript will undergo copyediting, typesetting, and review of the resulting proof before it is published in its final form. Please note that during the production process errors may be discovered which could affect the content, and all legal disclaimers that apply to the journal pertain.

ACCEPTED MANUSCRIPT 1 1

The occurrence and effect of unit operations for dairy products processing on the fate of

2

aflatoxin M1: a review

3 Fernanda Bovo Campagnollo1, Karina C. Ganev2, Amin Mousavi Khaneghah1, Jéssica Portella3,

5

Adriano G. Cruz3, Daniel Granato4, Carlos H. Corassin5, Carlos Augusto F. Oliveira5, Anderson

6

de Souza Sant’Ana1

RI PT

4

1

8

SC

7

Department of Food Science, Faculty of Food Engineering (FEA), University of Campinas

10

(UNICAMP), Campinas, SP – Brazil. 2

M AN U

9

Salvador Arena Foundation Educational Center, Thermomechanics Faculty of Technology, São

11

Bernardo do Campo, SP – Brazil. 3

4

13 14

17 18 19 20 21

4748, 84030-900, Ponta Grossa, Brazil.

5

Department of Food Engineering, Faculty of Animal Science and Food Engineering, University of São Paulo (USP), Pirassununga, SP – Brazil.

EP

16

Department of Food Engineering, State University of Ponta Grossa. Av. Carlos Cavalcanti,

AC C

15

Federal Institute of Science and Technology of Rio de Janeiro, Rio de Janeiro, RJ, Brazil.

TE D

12

Short title: Fate of aflatoxin in dairy processing

22

*Corresponding author: Prof. A.S.Sant’Ana: [email protected]

23

Address: Rua Monteiro Lobato, 80. Cidade Universitária Zeferino Vaz. CEP: 13083-862.

24

Campinas, SP, Brazil. Phone: +55(19) 3521-2174.

ACCEPTED MANUSCRIPT 2 Abstract

26

Mold contamination has challenged the safety of feed production and processing because of its

27

undeniable role in the spoilage and the possible consequent toxicity impact on human health and

28

the economy. Aflatoxin M1 (AFM1) is a hepatocarcinogenic derivative of aflatoxin B1 excreted

29

into the milk after ingestion of feed contaminated by certain molds. Because of the important role

30

of dairy products, especially milk in the human diet, there is a huge concern about the presence of

31

AFM1 in milk and dairy products. In this article, the occurrence of AFM1 and the fate of AFM1

32

during processing of milk and dairy products, such as yoghurt and cheeses, since 1996 until

33

today, was reviewed. The evaluation of mechanisms by which AFM1 is affected by each

34

processing step is of major importance to provide useful and accurate information to develop risk

35

assessment studies and risk management strategies.

M AN U

SC

RI PT

25

36

TE D

37 Keywords: Food safety; aflatoxins; mycotoxins; milk; food technology, dairy processing; 38 aflatoxin M1; cheese; yogurt; feed; Aspergillus; incidence; effect of processing; fermentation;

41 42 43 44 45 46 47 48

AC C

40

EP

39 thermal processing; salting; ripening; decontamination; detoxification.

ACCEPTED MANUSCRIPT 3 49 50

1) Introduction Milk and dairy products are commonly consumed by people of all age groups, especially

51 children. Accordingly, milk is one of the major nutrient sources. Milk is very important in human

RI PT

52 nutrition because of it is biochemical complexity and for providing all essential amino acids. 53 Confirmation of these nutritive benefits is the extensive and constant consumption of milk and 54 dairy products in several countries (Galvano et al., 1996). According to Silva et al. (2007), the

SC

55 consumption of dairy products constitutes more than 80% of the habits and dietary intake of 56 children worldwide.

Humans and animals are subjected to “biological hazard” from natural toxicants that occur

M AN U

57

58 in food and feed. Mycotoxins are secondary metabolites produced by a few fungal species 59 belonging mainly to the Aspergillus, Penicillium and Fusarium genera. Such compounds may be 60 formed by these mycotoxigenic molds when growing in contaminated foods at production,

TE D

61 processing, transportation, and also during storage. Aflatoxins, trichothecenes, zearalenone, 62 deoxynivalenol, fumonisin, patulin, ochratoxin, and ergotamine are the main mycotoxins that have 63 challenged the safety of feed production and food processing because they are negatively affecting

EP

64 human health and the economy (Murphy et al., 2006; Bhat et al., 2010). According to the Food and 65 Agriculture Organization of the United Nations (FAO), up to 25% of the world’s food crops are

67

AC C

66 significantly contaminated with mycotoxins. Aflatoxin is one of the most important mycotoxins and it can be produced by different

68 species of Aspergillus genus, mainly Aspergillus flavus and Aspergillus parasiticus (Elsanhoty et 69 al., 2014). The main economic source of this mycotoxin are cereal-based foods; however aflatoxin 70 can also be found in foods of animal origin such as milk and dairy products. According to Bhat et 71 al. (2010), at least 18 different types of aflatoxins have been identified, and aflatoxin B1 (AFB1), 72 B2 (AFB2), G1 (AFG1), G2 (AFG2), and M1 (AFM1) are the most important from a food safety

ACCEPTED MANUSCRIPT 4 73 standpoint. “B” (blue) and “G” (green) refer to the fluorescence colour observed under exposure of 74 mycotoxin to ultraviolet light, while “M” refers to an AFB1 metabolite found in milk or dairy 75 products (Murphy et al., 2006). The contamination of milk and dairy products with mycotoxin can

RI PT

76 occur by indirect contamination when lactating animals ingest AFB1 contaminated feed which will 77 pass to the milk as AFM1, and also by direct contamination, when molds can grow in milk (very 78 unlikely) or on dairy products as intentional additives or accidental contamination (Sengun et al.,

SC

79 2008). Therefore, milk and dairy products are particularly susceptible to contamination by AFM1 80 and are considered to pose certain risks for human health. Accordingly, milk has the greatest

M AN U

81 demonstrated potential for the introduction of aflatoxin residues in the human diet (Galvano et al., 82 1996). 83

As milk and dairy products are processed by different technologies involving various unit

84 operations and present diverse chemical compositions, the effects of each processing step on the

TE D

85 mycotoxin levels in the final product may be variable. Although some strategies have been 86 proposed to prevent, control, and/or reduce the incidence of aflatoxins in animal feed and food, it 87 is known that their effectiveness in reducing the levels of mycotoxins is limited, present high costs

EP

88 or lead to nutritional and sensory changes that are perceived as ‘unacceptable’ (El-Nezami et al., 89 1998). Given the above, the knowledge on the occurrence of AFM1 in dairy products, and how it is

AC C

90 affected by each processing step, is of major importance to provide useful and accurate 91 information for the development of risk assessment studies and risk management strategies. In this 92 scenario that bounders food safety, technology, and public health, this study aims to review the 93 incidence and the fate of AFM1 in milk, yoghurt and cheeses during processing. 94 95 96

ACCEPTED MANUSCRIPT 5 97 98

2) Aflatoxins: characteristics, producing fungi, regulations and incidence Aflatoxins are compounds that have strong effect on human and animal health because they

99 lead to serious damage to the liver, induction of tumors as well as immunosuppressive, mutagenic,

RI PT

100 teratogenic and carcinogenic effects (Hernandez-Mendoza et al., 2009). Aflatoxins are fungal 101 metabolites produced by at least 20 species of three different sections of the Aspergillus genus, 102 such as Flavi, Nidulantes and Ochraceorosei. The members included in the Flavi section are A.

SC

103 arachidicola, A. bombycis, A. flavus, A. minisclerotigenes, A. nomius, A. novoparasiticus, A. 104 parasiticus, A. parvisclerotigenus, A. pseudocaelatus, A. pseudonomius A. pseudotamarii, A.

M AN U

105 togoensis, A. transmontanensis, A. mottae and A. sergii; the members in the Ochraceorosei section 106 are A. ocharaceroseseus and A. rambelii; and finally the members inserted in the Nidulantes 107 section are A. astellatus, A. olivicola and A. venezuelensis (Varga et al., 2009; Baranyi et al., 2013; 108 Baranyi et al., 2015). Although all these species are reported as aflatoxin producers, in which A.

TE D

109 flavus and A. parasiticus are the most notorious. Generally, aflatoxins are produced in sub-optimal 110 temperatures for the growth of the species involved. Studies regarding A. flavus have shown that 111 the optimum temperature for its growth is between 29°C and 35°C, with maximum aflatoxin

EP

112 production at 24ºC, and no production occurs at temperatures below 13°C or above 42°C. The 113 maximum production in culture medium is related to the depletion of fermentable carbohydrates

AC C

114 and secondary metabolism; some researchers reported that the presence of barium in the culture 115 medium inhibits the mycotoxin production (Baptista et al., 2004). 116

Formation of mycotoxins is not a continuous process, but it must be assumed that if a

117 mould is present in the environment and is capable of producing toxins, the mycotoxins will be 118 present in the food. In addition, the fungus may be absent, but the toxin may be present and active 119 (Ferreira et al., 2006). Fungal multiplication and aflatoxin production are determined by the 120 chemical composition of the substrate, its water content, and environmental conditions, such as

ACCEPTED MANUSCRIPT 6 121 temperature and humidity (Jay, 2005). Bryden (2012) explains that other factors can also influence 122 the quantity of aflatoxin produced, such as mechanical damages, the presence of carbon dioxide 123 and oxygen, application of pesticides and fungicides, plant variety, insect infestation and amount

RI PT

124 of spores. The level of contamination is cumulative, so the time to harvest and the drying and 125 storage conditions can play an important role in the production of aflatoxin (Prandini et al., 2009). 126

Aflatoxins are soluble in solvents such as methanol, chloroform and benzene, with a wide

SC

127 spectrum of toxicity and have a low molecular weight. They are non-immunogenic, act at low 128 concentrations and are unstable at UV light, but very stable at temperatures above 100 °C,

M AN U

129 presenting small or almost no decomposition when subjected to baking, roasting and pasteurization 130 (Midio and Martins, 2000; Ferreira et al., 2006). Aflatoxins have a polycyclic structure derived 131 from a coumarin nucleus linked to a bifurano system, while aflatoxins of type B are connected to a 132 pentanone, G-type aflatoxins are connected to a 6-membered lactone (Abrar et al., 2013). The

TE D

133 toxicity of aflatoxins in animals is as diverse as the fungal species that produce these compounds. 134 In addition of the acute toxicity, the incidence of certain types of cancer has been associated to 135 some mycotoxins and this aspect has drawn attention to the feed and food safety, especially for

137

EP

136 milk and dairy products (Castegnaro and Mcgregor, 1998). According to Santini and Ritieni (2013), aflatoxins are considered lipophilic molecules, and

AC C

138 as the liver is a predominantely lipophilic organ, all compounds which are transported by the blood 139 stream are stored and concentrated in the hepatocytes. Herein, these compounds can induce some 140 types cancer if the organ is continuously exposed to aflatoxins. After ingestion, AFB1 is 141 biodegraded in the liver by the cytochrome P450 enzymatic system. The main reactions involved 142 in AFB1 metabolism are: hydroxilation, oxidation and demethylation. AFB1-8,9-epoxide (AFBO) 143 is a highly toxic, mutagenic and carginogenic product. AFM1 is also highly toxic whereas aflatoxin

ACCEPTED MANUSCRIPT 7 144 P1, Q1 or B2a are relatively nontoxic and are formed during these metabolic pathways (Wu et al., 145 2009). The main reactions involved in metabolism of AFB1 are described in Figure 1. 146 Insert Figure 1 here.

RI PT

147 148 149

After being biodegraded, AFBO can be bound to cellular macromolecules, including the

SC

150 genetic material, such as proteins and DNA, forming adducts with nucleic acids (Murphy et al., 151 2006). These adducts are converted into a stable open ring derived from formamidopyrimidine,

M AN U

152 and the repair of these lesions leads to the appearance of genetic mutations and cancer. 153 Hydroxylated metabolites and other aflatoxins that occur naturally are not suitable substrates for 154 the epoxidation reaction and thus are less mutagenic and carcinogenic (Bartoszek, 2006). The 155 excretion of some of these compounds in the urine of infected individuals is used as an evidence

TE D

156 that humans have the biochemical pathways necessary for carcinogenesis, and also provides a 157 reliable biomarker for exposure to AFB1 (Murphy et al., 2006). 158

When livestock, including dairy cattle, ingest AFB1 through the consumption of

EP

159 contaminated feed, a portion of this mycotoxin is degraded in the rumen by resident 160 microorganisms resulting in the formation of aflatoxicol. The AFB1 remaining part is absorbed in

AC C

161 the digestive tract by passive diffusion (Fink-Gremmels, 2008) and undergoes hepatic 162 biotransformation becoming AFM1, an hydroxylated form of AFB1 which is excreted in milk, 163 tissues and biological fluids of these animals (Oatley et al., 2000; Peltonem et al., 2001; Murphy et 164 al., 2006). AFM1 structure is a 4-hydroxy form of AFB1 (C17H12O7). Additionally, aflatoxin M2 165 (AFM2) is a 4-dihydroxy form of AFB2 (C17H14O7). The structures of the most common aflatoxins 166 can be seen in Figure 2 (Henry et al., 2001).

ACCEPTED MANUSCRIPT 8 167

Creppy (2002) reported that approximately 0.3% to 6.2% of the total AFB1 ingested by the

168 animals are usually transformed into AFM1 in the milk. Bakirci (2001) stated that there is a linear 169 relationship between the amount of AFM1 in milk and the AFB1 contaminated feed consumed by

RI PT

170 the cows. According to Battacone et al. (2005), the carry-over rate in cows varies from 0.35% to 171 3%, and in sheep this rate varies from 0.08% to 0.33%. Additionally, Masoero et al. (2007) 172 observed that the total AFM1 excretion and its carry-over in milk were affected by the milk yield,

SC

173 in which high-milk-yield cows present higher AFM1 excretion as compared to low-milk-yield 174 cows. These authors also concluded that the variability observed in the AFM1 carry-over among

M AN U

175 animals could be related to differences in rumen degradation activity, in aflatoxicol formation, in 176 the enzymatic AFB1 oxidation system, and in the permeability of mammary glands. Excretion of AFM1 in milk starts after 12-24 h ingestion of contaminated feed, reaching

178

high levels in a few days and disappearing approximately 24 h after it was eliminated from the

179

diet (Nachtmann et al., 2007). The occurrence of AFM1 in milk is influenced by numerous

180

factors, such as animal species, variability of individuals, lactation, milking, animal’s and udder

181

health, feed intake, level of contamination, geographic location and season of the year. Therefore,

182

aflatoxin absorption rate and excretion of AFM1 in milk vary intra-animal, intra and inter-day and

183

milking (Fink-Gremmels, 2008; Virdis et al., 2008).

EP

TE D

177

Prandini et al. (2009) explain that cows that ingested an amount of AFB1 less than 40

185

µg/cow/day produce milk with AFM1 content of less than 0.05 µg/kg. Also, AFM1 production

186

reactions are very fast, since AFM1 appears in milk 2 to 3 days after eating contaminated feed, as

187

well as AFM1 level in milk is reduced to zero in 2 to 3 days after the animal is fed with a diet

188

without aflatoxins. A study carried out by Battacone et al. (2005) showed that AFM1 was

189

detected in sheep milk after 12 h of toxin intake. On the seventh day, the administration of

AC C

184

ACCEPTED MANUSCRIPT 9 190

contaminated feed was interrupted and the AFM1 average concentrations declined rapidly and,

191

after four days, AFM1 was not detected in any milk sample.

193

2.1) International regulations for AFM1

RI PT

192

The International Agency for Research on Cancer (IARC, 2002) considered AFM1 as

195

belonging to Group 1 (carcinogenic to humans). Because of the high incidence of this mycotoxin

196

in milk and dairy products, its presence in food can be considered an issue of major relevance for

197

public health. According to Galvano et al. (1996) and Prandini et al. (2009), milk shows the

198

greatest potential for the introduction of aflatoxin in human nutrition. Furthermore, milk contains

199

the major nutrients for the growth of children, whose remarkable and potentially sensitivity is

200

higher than in adults. Silva et al. (2007) explained that the consumption of dairy products

201

constitutes more than 80% of the habits and dietary intake of children worldwide.

M AN U

According to van Egmond et al. (2007), different factors can influence the setting of limits

TE D

202

SC

194

203 for mycotoxins, including the availability of toxicological and occurrence data, detailed knowledge 204 about the distribution and sampling possibilities, and the analytical methods for raw materials and

EP

205 final products. Complimentarially, socio-economic issues, such as the existence of regulations in 206 the countries where the commercial trade will be held, and the need of sufficient demand for food

AC C

207 are also noteworthy. Therefore, in order to protect the public health, several countries have 208 established food safety standards for maximum acceptable levels of aflatoxin present in food and 209 feed, especially in milk and dairy products. In this scenario, Galvano et al. (2001) reported that 210 some regulatory limits in several countries are influenced by economic considerations and 211 generally have little or no scientific basis. In contrast, Zinedine and Mañes (2009) found that the 212 decision limits were based on scientific evidences from risk assessment (toxicological data), food

ACCEPTED MANUSCRIPT 10 213 consumption data, and detailed knowledge about the possibilities of sampling and analysis, and 214 socio-economic issues. 215

Several countries adopt a strict legislation for AFM1 such as 50 ng/kg as the maximum level

RI PT

216 in the European Union and 500 ng/kg established by the United States of America (IARC, 2002). 217 In most developed countries, research on contamination levels has been carried out aiming at 218 checking contamination and generating community awareness (Souza et al., 1999). Approximately

SC

219 60 countries have already established regulatory limits for AFM1 in milk and dairy products 220 (Josephs et al., 2005). The US-FDA (United States Food and Drug Administration) has established

M AN U

221 AFM1 levels of 0.5 µg/kg in milk and 20 µg/kg in animal feed, while the Codex Alimentarius 222 Committee has recommended a maximum level of 0.05 µg/kg in milk (van Egmond and Jonker 223 2004; Jay, 2005). The European Union has set a maximum limit of 0.05 µg/kg in raw milk, heat224 treated milk and milk for the manufacture of milk-based products (van Egmond and Jonker 2004;

TE D

225 Murphy et al., 2006). Brazilian law adopts the MERCOSUL maximum permitted levels of 0.5 226 µg/kg in liquid milk, 5.0 µg/kg in milk powder and 2.5 µg/kg in cheese (Anvisa, 2011). According 227 to Var and Kabak (2009), there is a lack of agreement concerning the maximum AFM1 levels

EP

228 worldwide, being a problem for marketing of certain products, as they can be accepted in some 229 countries and not in others, depending on the degree of their development and economic status.

AC C

230 Table 1 shows the maximum permitted levels of AFM1 in milk and dairy products in some 231 countries. 232 233 234 235 236

Insert Table 1 here.

ACCEPTED MANUSCRIPT 11 237

2.2) Worldwide Incidence of AFM1

238 There are several studies on the occurrence of AFM1 in milk and dairy products, which

240

have been performed internationally, demonstrating the occurrence and consequent global

241

concern over this mycotoxin (Kamkar, 2005). This section will report the occurrence of AFM1 in

242

different types of bovine milk (pasteurized, raw, UHT) and dairy products, such as cheese and

243

yoghurt, as well as milk from other animal sources (sheep, goat, buffalo). The observations

244

indicate that the AFM1 contamination in milk and dairy products is mostly perceived in specific

245

geographical regions.

M AN U

SC

RI PT

239

Regarding the methodology for identification and quantification of AFM1, chromatographic

247

and immunochemical methods are generally used. According to Jalili and Scotter (2015), these

248

analytical methods are rapid, selective, sensitive (low limit of detection), reliable and cost

249

effective. Authors observed that the most used chromatographic methods (used for confirmation

250

of the results from rapid mycotoxin screening tests) are thin-layer chromatography (TLC) and

251

high performance liquid chromatography (HPLC). For immunochemical methods, rapid tests

252

(presence/absence of AFM1) based on specific antibodies, enzyme-linked immunosorbent assay

253

(ELISA) is the most used, but other methods, such as immunoaffinity column assays, sequential

254

injection immunoassay and radioimmunoassay, are also employed.

256 257

EP

AC C

255

TE D

246

2.2.1) Milk

It can be seen in Table 2 the incidence of AFM1 in different types of milk in some countries

258 around the world. According to Galvano et al. (1996), it is quite impossible to compare the 259 extracted results from the literatures because of the wide differences between and also within the 260 summarized countries. Some of factors responsible for such differences are: feeding producers,

ACCEPTED MANUSCRIPT 12 261 type of animals, environmental factors, such as dryness, seasonal variation, applied extraction and 262 analysis procedures and established regulatory limits for AFM1 for both of feed and milk

264

Insert Table 2 here.

265

RI PT

263

It can be seen in Table 2 that the number of samples contaminated with AFM1 is variable,

267

which can be justified by the sensitivity difference in the methodology used to quantify the

268

aflatoxin; in addition to the different levels of contamination of the feed (Sassahara et al., 2005).

269

Shundo et al. (2009) estimated the AFM1 exposure in milk and its occurrence in Sao Paulo,

270

Brazil. A high incidence of AFM1 in milk and dairy products with low levels of contamination

271

was observed, so the occurrence of AFM1 does not appear to be a serious health hazard,

272

considering the Brazilian legislation. In another study performed in Brazil, Oliveira et al. (2013a)

273

concluded that some factors, such as low availability of green fodder, excessive use of

274

concentrated feed, contamination of feed with aflatoxin during storage and also inappropriate

275

feeding of animals, contributed to the high level of AFM1 in milk. Sassahara et al. (2005)

276

explained that the differences presented regarding other studies can be justified by the sensitivity

277

difference among the used methodology.

M AN U

TE D

EP

Han et al. (2013) concluded that there were no differences in the AFM1 level of 200 milk

AC C

278

SC

266

279 samples from China (South and North regions). Tajkarimi et al. (2008) conducted a survey 280 regarding the aflatoxin levels of 319 raw milk samples from 14 different regions of Iran and 281 observed that the distribution of AFM1 was not the same within regions. Malissiova et al. (2013) 282 compared the contamination of organic and conventional ewe’s and goat’s milk. For this purpose, 283 234 samples were collected from organic and conventional farms and samples were screened for 284 AFM1, and organic milks had higher AFM1 levels.

ACCEPTED MANUSCRIPT 13 285

According to the results of Bilandžić et al. (2010), the contamination of milk samples with

286 AFM1 was significantly higher during winter-spring seasons in comparison with summer and 287 autumn. The authors also observed that 98.4% of the examined milk samples in Croatia contained

RI PT

288 AFM1 levels below the maximum limit approved by the European Union. In addition, Unusan 289 (2006) observed that the contamination of milk and dairy products in the summer was significantly 290 lower than in the winter and autumn. In a recent study of Picinin et al. (2013), 129 milk samples

SC

291 were collected from different dairy farms in three periods (dry, transition and rainy period), and it 292 was observed that the concentration of the toxin was significantly affected by climate condition,

M AN U

293 hence the highest level was found in the dry period. Fallah (2010b) investigated the AFM1 level of 294 298 dairy products collected in four urban Iranian cities and observed that the seasonal effect on 295 the occurrence and AFM1 level in pasteurized milk samples, yoghurt, butter and ice cream 296 collected in the winter were significantly higher (P<0.05) than the samples collected in the

TE D

297 summer. Bilandžić et al. (2014b) analysed the AFM1 content of 3716 samples of raw milk and 706 298 samples of UHT milk commercialized in Croatia and the incidence of AFM1 contamination was 299 lower in the summer. Authors suggested that this occurs because the producers use raw feed as

301

EP

300 pasture instead of concentrated feed.

Nakajima et al. (2004) analysed the level of contamination of 208 commercial pasteurized

AC C

302 milk samples collected in retail outlets in Japan during the winter and they concluded that the mean 303 AFM1 concentration in Japanese milk samples was similar to the concentration contained in the 304 diet of Middle East people and also lower than those levels found in European, Latin Americans 305 and Far Eastern diets. El Marnissi et al. (2012) recommended that more studies regarding the milk 306 contamination associated with observations on feeding producers are needed to find important 307 factors involved in the AFM1 contamination. Strict controls of raw materials and feeds, especially 308 those using green fodder for cattle feeding, have been recommended by Hussain et al. (2008) and

ACCEPTED MANUSCRIPT 14 309 Iqbal et al. (2011). In South Africa, Dutton et al. (2012) analysed 45 milk samples and observed 310 AFM1 levels varying from 0.01 to 3.1 µg/L. The obtained data confirmed that the average AFM1 311 level (0.278 µg/L) of milk samples collected in the winter was higher than the milk samples

313

RI PT

312 produced in the summer (0.022 µg/L).

As can be seen in the studies analysed, many authors have shown the influence of seasonal

314 effect on the level of AFM1. On average, the winter milk samples revealed higher AFM1

SC

315 concentrations than the summer samples. Hence, countries located in colder regions also present 316 higher levels of AFM1 in milk samples. During the winter season, the availability of fresh animal

M AN U

317 feed such as pasture, grass and green fodder is reduced and the producers change the feeding 318 practices and appeal to the usage of concentrated feedstuff. This kind of feed is generally 319 composed by corn, wheat and cotton seeds that could be stored under inadequate conditions and 320 may contain toxigenic fungi like Aspergillus and consequently be contaminated with high levels of

TE D

321 aflatoxins. In addition, there is evidence that milk yield is lower in winter, leading to increase in 322 the concentration of AFM1 and other components (Asi et al., 2012). 323

It is very important to implement preventive programs to reduce the AFM1 occurrence, as

EP

324 well as strict regulations by government to control and minimize its impact on all types of dairy 325 products. Generally, studies indicate that aflatoxins are not a risk to consumer’s health because

AC C

326 although the incidence of AFM1 is relatively high, the levels are below the regulatory limits of 327 each country. However, if the effects associated with the high consumption of contaminated 328 products in the long term are combined with the high exposure, and incidence of cases were taken 329 into account, the contamination by mycotoxins may become one of the most serious public health 330 problems. 331 332

ACCEPTED MANUSCRIPT 15 333 334

2.2.2) Other Dairy products The incidence of AFM1 in different dairy products, such as cheeses, yoghurts, butter, ice

336 337

Insert Table 2 here.

338

Fernandes et al. (2012) studied the distribution and stability of AFM1 in Minas Frescal

SC

339

RI PT

335 creams, milk powder and milk cream, is summarized in Table 2.

340 cheese manufactured with or without starter cultures. Authors observed that the storage time had

M AN U

341 no effect on the AFM1 content and the milk containing high AFM1 level concentrated the toxin in 342 Minas Frescal cheese. Furthermore, adding starter cultures did not influence the concentration or 343 even the stability of the toxin throughout 30 days of storage. The results of Nilchian and Rahimi 344 (2012), Bakirci (2001) and Deveci (2007) showed that the increase of AFM1 levels in the cheese is

TE D

345 a function of cheese type, type of unit operations and the amount of eliminated water during 346 processing. In addition, there are some specific factors that can affect the level of AFM1 in the 347 cheese curd, such as renneting temperature, duration of pressing, and pH of the saturated brine. Complimentarially to those experimental observations, variation in the contents of AFM1 in

EP

348

349 different types of cheese may be a result of several other factors, such as heat treatment,

AC C

350 proteolysis, and exposure of contaminated milk to light (Yousef and Marth 1989). Fallah et al. 351 (2009) and Mohajeri et al. (2013) observed that the variations in AFM1 content in cheese are also 352 due to the analytical method used to quantify AFM1. 353

In a report by Montagna et al. (2008), 256 cheese samples made from bovine, buffalo,

354 sheep, sheep-goat mixed milks were analysed for AFM1. The authors observed that AFM1 was 355 detected in 16.6% of the cheeses tested and cheeses made of goat and sheep milk were negative. 356 The results showed that the amount of AFM1 in milk from goats and sheep is less than in cows’

ACCEPTED MANUSCRIPT 16 357 milk, and this may be because of the differences in their digestive apparatuses and mechanism of 358 AFB1 assimilation in animals, and for the different patterns of feeding. In other words, cattle 359 fodders are more susceptible to be contaminated by AFB1 than those used to feed sheep and goats

RI PT

360 (Fallah et al., 2009; Fallah et al., 2011). Anfossi et al. (2012) also reported that Italian cheeses 361 produced with goat’s and sheep’s milks are less contaminated with AFM1 than cheeses made with 362 cow’s milk.

According to Manetta et al. (2009), there is a direct correlation between the AFM1 level

SC

363

364 found in milk and the level found in the final products. Anfossi et al. (2012) explained that

M AN U

365 industrial scale products contain less levels of AFM1 than artisanal products, which can be justified 366 by the fact that artisans often use just one milk source which can occasionally be contaminated 367 with high levels of AFM1, but industries use a combination of milk from different sources, so the 368 risk of contamination is lower. The authors also commented that the maturation stage could

TE D

369 decrease the contamination level probably because of degradation of the toxin, but several authors 370 mentioned that this stage does not significantly alter the toxin concentration. 371

Ardic et al. (2009) concluded that cheese is a potential source of AFM1 as compared to

EP

372 other dairy products because this toxin is associated to the casein fraction (which is concentrated in 373 cheese). Some studies showed that the AFM1 level, in some types of soft cheese, is about 3-fold

AC C

374 higher and about 5-fold higher in hard cheese compared to the milk used in the manufacture 375 (Bakirci, 2001; Deveci, 2007; Kamkar et al., 2008; Prandini et al., 2009). In contrast, the results 376 from Elgerbi et al. (2004) indicated that AFM1 level in cheese products were lower than in the raw 377 milk (Govaris et al., 2001; Deveci, 2007; Kamkar et al., 2008). 378 379 380

ACCEPTED MANUSCRIPT 17 381

3) The Fate of AFM1 during Milk Processing Milk is an important nutritious food but it has a short shelf life, requiring a rigorous

383

manipulation in order to avoid its spoilage and the transmission of diseases to consumers. The

384

processing of milk ensures its conservation for days or for a longer time depending on the type of

385

transformation (cooling, heating, fermentation, etc.), besides the production of different products

386

that are appreciated by consumers, such as cheeses, yoghurts, and butter. Dairy products differ

387

significantly among countries, depending on dietary habits and tradition, accessible technologies

388

for milk processing, market requirement, and social and cultural status. Figure 3 illustrates the

389

basic processing of pasteurized and UHT milk. UHT milk management has the same production

390

steps as those of pasteurized milk until the standardized milk is composed.

M AN U

SC

RI PT

382

391 392

Insert Figure 3 here.

394

TE D

393 3.1) Effects of pre-processing steps

From Figure 3, depicting the main steps applied for milk processing, it is clear that the

396

source of AFM1 is the raw milk. Thus, the presence of aflatoxin in the feed and its transformation

397

into AFM1 are important aspects to be taken into consideration. Feed contamination by aflatoxin

398

is associated with high temperatures and an extended drought, and is often problematic in warm,

399

humid, tropical and subtropical crop-growing regions (Payne, 1998). According to Bryden

400

(2012), the production of aflatoxins is not specifically restricted to any ingredient of the animal

401

feeding, but the degree of contamination by aflatoxins varies with location and climatic

402

conditions. Gizachew et al. (2016) analysed the AFB1 contamination in 156 dairy feed samples in

403

Ethiopia and observed that all samples were contaminated with AFB1 (7 to 419 µg/kg). The

404

collected feed samples included concentrated feed (composed by wheat bran, noug cake, pea

AC C

EP

395

ACCEPTED MANUSCRIPT 18 hulls and maize grain), brewery by-product, maize grain, pea hulls and silage. Authors concluded

406

that there was a moderate positive correlation between AFB1 in feed and AFM1 in milk (r=0.31).

407

The analysis of individual feeds revealed that noug cake was the main contributor for the AFB1

408

contamination. Pleadin et al. (2015) studied different feed samples (maize, wheat, barley, oat,

409

grain mixtures and manufactured dairy feed) in Croatia and concluded that maize was the most

410

contaminated with AFB1 (31.4%), followed by grain mixture (26.2%) and dairy cattle feed

411

(22.2%), which presented 12.3% of samples surpassing the AFB1 limit permitted by the European

412

Union (5 µg/kg). The authors stated that the high levels of AFB1 contamination can be attributed

413

to extreme warm and drought weather during growth and harvesting period. In Spain, Hernández-

414

Martínez and Navarro-Blasco (2015) observed that 70 out of 78 samples of dairy cow feedstuff

415

contained detectable levels of aflatoxin, but none of the samples exceeded the maximum AFB1

416

level recommended by the European Union. The researchers also concluded that the geographical

417

location, unlike season or feeding system, had limited influence on aflatoxin levels. Similarly, in

418

the study performed by Simas et al. (2007) in the State of Bahia, Brazil, aflatoxins (1 to 3 µg/kg)

419

were found in 33.8% of 80 samples of brewers grain used to feed dairy cattle. A positive

420

correlation was observed between the aflatoxin level, mean rainfall and relative humidity,

421

indicating that these factors have an expressive influence on the moisture content. Likewise, in

422

China, Han et al. (2013) verified that 42% out of 200 feed samples were positive for AFB1 (0.05

423

to 3.53 µg/kg). Furthermore, there was no difference in the contents of aflatoxins between the

424

north and the south of the country. Summarizing these results, it is possible to infer that the

425

aflatoxin contamination in dairy feed is a worldwide problem.

AC C

EP

TE D

M AN U

SC

RI PT

405

426

As aflatoxin is present in feeds and is consumed by livestock, it can be transformed into

427

AFM1. Studies have shown a considerable variability concerning the percentage of aflatoxins

428

transformed into AFM1 and the amount of this mycotoxin present in milk. According to Prado et

ACCEPTED MANUSCRIPT 19 al. (1999), the relationship between aflatoxin intake and excretion in milk is highly variable,

430

averaging in 1.5%. Battacone et al. (2003) observed that AFM1 level in ewe’s milk was

431

significantly affected by the AFB1 dose. Moreover, AFM1 level increased linearly as the AFB1

432

intake increased. However, the carry-over (average of 0.112%) was not associated to the quantity

433

of AFB1 ingested by the animals. Hussein and Brasel (2001) indicated that the biotransformation

434

of AFB1 into AFM1 was comprehended in the 0.5-5% range. Lindner (1995) reported that at least

435

2.2% of AFB1 is excreted as AFM1 in cow's milk. For other animal species, the AFM1 excreted in

436

goat milk is between 0.18% and 3% of the amount of ingested AFB1 (Virdis et al., 2008), while

437

in sheep milk this value ranges from 0.08% to 0.33%, which are lower than the values found in

438

cow’s milk, 0.35% to 3% (Battacone et al., 2005). According to Chopra et al. (1999), some

439

factors can influence the AFM1 level in milk, such as milk yield, microsomal mixed function

440

oxidase activity and the presence or absence of bacterial mastitis in the udder. In addition, carry-

441

over is influenced by species, individual variability, lactation stage and milking (Virdis et al.,

442

2008).

TE D

M AN U

SC

RI PT

429

The variability in AFM1 presence in milk reinforces that strategies must be implemented to

444

reduce the production and the occurrence of aflatoxins in feeding materials. Nonetheless, if

445

present in feeds, management programs should be implemented in order to reduce animal

446

exposure to aflatoxin (Pereira et al., 2005). According to Oliveira and Ferraz (2007), in some

447

countries, the care about the quality of milk from other species is even lower than that for cow's

448

milk. To reduce milk contamination, it is necessary to reduce AFB1 grain contamination, to select

449

varieties resistant to toxigenic fungi, to prevent physical damage by insects, and to perform

450

appropriate crop rotation. In addition, grains considered improper for human consumption should

451

not be offered to lactating animals. Drying is a critical step prior to storage, and provides ideal

452

conditions of temperature, low moisture and aeration (Beltrane and Machinski, 2005). In case of

AC C

EP

443

ACCEPTED MANUSCRIPT 20 failures in preventive measures, aflatoxins can be reduced by either mixing the contaminated

454

material with foods that have lower (or no) concentrations or by chemical, physical or biological

455

treatments (Oveisi et al., 2007). Phycal processes emcompass thermal inactivation, application of

456

ultraviolet light and/or ionizing radiation, while chemical methods employ solvent extraction,

457

break mycotoxin structures, such as chlorine treatments (sodium hypochlorite and chlorine gas),

458

oxidizing agents (hydrogen peroxide, ozone and sodium bisulfite) or hydrolytic agents (acids,

459

alkalis and ammonia). Nevertheless, both physical and chemical methods present disadvantages,

460

such as inefficient removal, high costs or nutritional losses. Biological methods elapse from the

461

action of microorganisms including yeasts, filamentous fungi, bacteria and algae, which act on

462

mycotoxins through mechanisms based on competition for nutrients and space, interactions and

463

antibiosis (Oliveira et al., 2013b).

M AN U

SC

RI PT

453

Biological decontamination methods can be a very promising choice, as they are efficient,

465

specific, practical, large-scale and cost-effective (Rahaie et al., 2012). Lactic acid bacteria (LAB)

466

and yeasts represent prominent groups due to their wide use in fermentation and preservation of

467

food. Shetty and Jespersen (2006) reported that LAB and yeasts have a high capacity for removal

468

of mycotoxins and can be used as part of starter cultures in fermented foods and beverages,

469

presenting fermentative and decontaminant capacity, or purified components of these same

470

strains can be used in small quantities as food additives without compromising the final product

471

characteristics. Thus, cells of these microorganisms added to the feed of dairy cattle can bind to

472

aflatoxins and consequently avoid the biotransformation of AFB1 into AFM1, reducing the

473

toxicological hazards that aflatoxins can cause to humans and animals. Studies in vivo

474

considering the effects of the addition of LAB in feed are scarce, but in vitro studies are vastly

475

available. El-Nezami et al. (1998) evaluated the ability of seven strains of LAB in binding the

476

AFB1 and observed that Lactobacillus rhamnosus (strains GG and LC-705) bound 80% of AFB1.

AC C

EP

TE D

464

ACCEPTED MANUSCRIPT 21 Haskard et al. (2001) analysed nine strains of Lactobacillus and also concluded that the GG and

478

LC-705 strains were more efficient in binding AFB1, reaching 78.9% and 76.5%, respectively.

479

Peltonem et al. (2001) studied 15 LAB strains of Lactobacillus and Lactococcus, and five strains

480

of bifidobacteria, and obtained AFB1 binding results between 5.6% and 59.7%. Lactobacillus

481

amylovorus (strains CSCC5160 and CSCC5197) and L. rhamnosus LC1/3 presented the best

482

results, 59.7%, 57.8% and 54.6%, respectively. Oatley et al. (2000) observed that different strains

483

of bifidobacteria bound 37% to 46% of AFB1. It can concluded that within a genus and/or

484

species, not all strains are equivalent in terms of aflatoxin binding; on the contrary, this ability is

485

characteristic of specific strains with the effectiveness varying markedly (El-Nezami et al., 2004).

486

As for LAB, yeast cells (mainly Saccharomyces cerevisiae) have been studied for their

487

capacity of AFB1 binding. Joannis-Cassan et al. (2011) analysed products containing S.

488

cerevisiae (cell wall of baker or brewer yeast, baker inactive yeast and brewer yeast) and

489

observed that AFB1 binding activity varied from 2.5% to 49.3%, depending on the toxin

490

concentration and the product type. Shetty et al. (2007) reached similar results for a S. cerevisiae

491

strain, 69.1% binding when AFB1 concentration in the medium was 1 µg/mL, 41% binding at 5

492

µg/mL and 34% binding at 20 µg/mL. Bovo et al. (2015) studied the capacity of products derived

493

from S. cerevisiae cells (differing in the viability and integrity of cells) in binding AFB1 at

494

different pHs (4.0 and 6.0). AFB1 adsorption results ranged from 45.5% to 69.4% at pH 3.0 and

495

from 24.0% to 63.8% at pH 6.0, and the higher percentage of AFB1 binding (P<0.05) at both pHs

496

was achieved when the products containing hydrolyzed yeast cells or yeast cell walls instead of

497

intact yeast cells were used.

AC C

EP

TE D

M AN U

SC

RI PT

477

498

In addition to the use of microbial cells (viable or not) aiming to bind and adsorb aflatoxins,

499

studies have also been conducted on the use of sorbents such as clay materials to reduce total

500

aflatoxin (B1 + B2 + G1 + G2) in milk (Applebaum and Marth, 1982b, Carraro et al., 2014, Galo

ACCEPTED MANUSCRIPT 22 et al., 2010, Masoero et al., 2010, Natale et al., 2009). This is mainly done through the addition of

502

clay materials, such as hydrated aluminosilicate of sodium and calcium, sodium bentonite,

503

esterified glucomannan, sodium montmorillonite, diatomaceous earth, activated charcoal, etc., to

504

animal feed (Huwig et a., 2001, Piva et al., 1995). These material can reduce the toxin

505

bioavailability by enteroadsorption, avoiding the aflatoxin adsorption in the gastrointestinal tract

506

and preventing its distribution to the target organ. Consequently, milk produced by the animals

507

will contain less AFM1 because of the lower absorption of AFB1 by their organism. In addition,

508

clay materials have also been tested for their effectiveness to remove aflatoxin directly from the

509

milk. Data has indicated that the removal efficiency of AFM1 present in milk by adding clay

510

materials directly to the milk is usually higher than 65% (Applebaum and Marth, 1982b, Carraro

511

et al., 2014, Natale et al., 2009). These findings indicate that the use of clay materials either in the

512

feed or in the milk can constitute an important strategy for detoxification of AFM1 content of

513

milk. As such, more studies should be done, particularly focused on the addition of clay and

514

binding materials in milk and the effects on nutritional and technological properties of dairy

515

products.

517

SC

M AN U

TE D

EP

516

RI PT

501

3.2) Effects of thermal treatments on AFM1 content The fate of AFM1 is variable and depends on the unit operations used for milk processing

519

(i.e., pasteurization, sterilization or spray drying) (Galvano et al., 1996). Despite this, studies

520

have shown that AFM1 is relatively stable to drying and thermal processing because the

521

decomposition temperatures of aflatoxins are between 237 to 306 oC (Rustom, 1997). Thus, as

522

raw milk is contaminated by AFM1, this mycotoxin will likely be found in the final product.

523

However, still there are some conflicting results: Awasthi et al. (2012) stated that pasteurization

524

process does not influence the level of AFM1 in bovine milk and the boiling of milk in household

AC C

518

ACCEPTED MANUSCRIPT 23 conditions is not able to decrease the concentration of AFM1 in milk. Jasutiene et al. (2006)

526

claimed that the heating (95 °C/3 min) did not influence the content of AFM1. Likewise, Govaris

527

et al. (2002) observed that the pasteurization (92oC/3 min) did not cause any significant

528

difference (P>0.01) in the AFM1 content. Conversely, Deveci (2007) reached reductions between

529

12 and 9% in pasteurized milk (72oC/2 min) artificially contaminated with 1.5 µg/L and 3.5 µg/L,

530

respectively. Bakirci (2001) observed a 7.62% decrease (P>0.05) in the AFM1 level after

531

pasteurization. Şanli et al. (2012) obtained reductions of 17.9% and 16.1% in AFM1

532

contamination after pasteurization (95 oC/5 min) of milk artificially contaminated at 1.5 and 3.5

533

µg/kg, respectively. Purchase et al. (1972) pasteurized milk using different conditions (62 oC/30

534

min, 72 oC/45 s and 80 oC/45 s) and obtained reductions of 32.5%, 45.5% and 63.6% in the

535

AFM1 level, respectively. When sterilization (115 oC/45 s) was applied, 81.3% of AFM1 was

536

reduced. El-Deeb et al. (1992) obtained 9.5% decrease in AFM1 content after heat treatment (63

537

°C/30 min) and 26% decrease after heating at 121°C for 15 min. According to Rustom (1997),

538

these contradictions could be attributed to the fact that there are differences in the initial level of

539

contamination, in the range of temperature and in the analytical methods used to extract, clean-up

540

and quantify the toxin. Moreover, the status of contamination can influence the results because in

541

artificially contaminated milk it is easier to inactivate AFM1 than in naturally contaminated milk.

543

SC

M AN U

TE D

EP

AC C

542

RI PT

525

3.3) Effects of concentration and drying

544

Not only thermal processing but also other steps impair some minor effects on AFM1, such

545

as milk concentration and drying process. A 35-40% reduction in the AFM1 level in pasteurized

546

skim milk samples was observed as a result of the concentration (30-33% total dry matter)

547

(Deveci and Sezgin, 2006). In addition, authors showed that when skim milk was spray-dried, the

548

total AFM1 contents were reduced about 59-68% compared to the original raw milk. AFM1

ACCEPTED MANUSCRIPT 24 distribution in milk is not homogeneous, 80% of milk protein is partitioned in the skim milk

550

portion connected to casein. Thus, AFM1 predominate in the nonfat fraction, which can be

551

explained due to its semipolar character (Galvano et al., 1996; Prandini et al., 2009). Heat

552

treatment is possibly the main cause of AFM1 reduction during the drying process because it can

553

cause the decomposition of milk proteins and the solubility of salts, and consequently the

554

hydrophobic interactions between AFM1 and casein are changed (Deveci and Sezgin, 2006).

555

Purchase et al. (1972) observed a reduction of 61%, 75.6% and 86.5% in milk dried by roller

556

drying under reduced pressure, roller drying and spray drying, respectively. Despite this, Galvano

557

et al. (1996) reported that some authors found no reduction in AFM1 toxicity in response to the

558

drying process, while other researchers observed reductions in the range of 60 to 75% according

559

to its concentration in milk. Nonetheless, it should be clear that with the reduction of water

560

levels, the transformation of fluid milk into powder will result in great increases in AFM1

561

concentration (Midio and Martins, 2000; Virdis et al., 2008).

563

SC

M AN U

TE D

562

RI PT

549

4) The fate of AFM1 during Cheese Processing A basic flowchart for cheese production can be seen in Figure 4. Standardized milk is

565

achieved after filtration, cooling, centrifugation and standardization of raw milk (level of fat in

566

the milk) as described in Figure 3. Worldwide, there are different types of cheeses, in which the

567

manufacture operations, types of milks (raw and pasteurized; bovine, caprine, ovine, and so on),

568

maturation periods (ripened and fresh/frescal cheeses), addition of herbs and other extracts,

569

among other factors differ between cheeses.

AC C

EP

564

570 571 572

Insert Figure 4 here.

ACCEPTED MANUSCRIPT 25 In general, cheese is an excellent substrate for fungal growth, but it cannot be considered a

574

suitable substrate for mycotoxin production (Bullerman, 1981). The occurrence of AFM1 in

575

cheese occurs mainly because of the use of contaminated milk. Researchers have reported that the

576

presence of AFM1 in cheese varies according to the type of product (Prado et al., 2001). Studies

577

have shown variable reductions of AFM1 during cheese making, ranging from 15% up to 65%. It

578

is known that the level of AFM1 in cheese is a function of different factors, including: cheese

579

type, technology strategies adopted in the manufacture process, amount of water removed during

580

processing, pH of saturated brine, cut size, renneting temperature, press time, contamination

581

degree of milk, differences in milk quality, and also depends on the analytical methods employed

582

in the quantification (van Egmond et al., 1977; Brackett and Marth, 1982a,b,c; Brackett et al.,

583

1982; Galvano et al., 1996; Oruc et al., 2006; Deveci, 2007; Kamkar et al., 2008; Sengun et al.,

584

2008; Ardic et al., 2009; Mohammadi et al., 2009; Fernandes et al., 2012; Motawee, 2013). The

585

fate of AFM1 contamination in the initial steps, such as in the pre-processing and thermal-

586

treatments, were described in the previous section. In this section, the fate of AFM1 in the

587

subsequent steps of cheese making will be discussed.

589

SC

M AN U

TE D

EP

588

RI PT

573

4.1) Effect of coagulation and draining on AFM1 content Coagulation and draining will be considered together because the curd and the whey are the

591

products resulted from these steps. Some studies have been conducted concerning the fate of

592

AFM1 in different types of cheeses. The carry-over of AFM1 from milk to Minas Frescal cheese,

593

the most popular cheese consumed in Brazil, ranged from 30.6% to 42.3% (Fernandes et al.,

594

2012). López et al. (2001) manufactured cheese using artificially AFM1 contaminated milk and

595

observed that the AFM1 distribution reduced about 60% and 40% in the cheese and whey,

596

respectively, compared to the original milk. Similarly, Colak (2007) obtained a mean AFM1

AC C

590

ACCEPTED MANUSCRIPT 26 reduction of 57.1% in Turkish white cheese and 65.3% in Kashar cheese, while the reduction in

598

the whey was 55.4%. Brackett and Marth (1982b) produced cheddar cheese from AFM1 naturally

599

contaminated milk and observed a 4.3-fold increase in AFM1 concentration in the curd, showing

600

that about 45% of the total toxin remained in the final product. The distribution and stability of

601

AFM1 during processing of traditional white-pickled cheese in Turkey was evaluated and the

602

AFM1 concentration in the curd was 3.6, 3.8 and 4.0-fold higher for different trial groups (50,

603

250 and 750 ng/L AFM1, respectively) compared to the raw milk. The authors complemented that

604

nearly half of the initial AFM1 spiked was found in cheese curd whereas the rest was drained by

605

the whey and a smaller part moved to the brine solution (Oruc et al., 2006). Brackett and Marth

606

(1982c) analysed the fate of AFM1 in parmesan and mozzarella cheeses. Parmesan presented a

607

5.8-fold AFM1 level compared to the initial naturally contaminated milk, while in Mozarella

608

cheese, a 8.1-fold increase was observed. The whey retained 20% of the concentration of AFM1

609

in milk. Cavallarin et al. (2014) observed that AFM1 content in the whey was between 30% and

610

65% compared to the original milk, and the production of Primosale, Robiola and Maccagno

611

cheeses showed AFM1 concentrations of 1.4, 2.2 and 6.7-fold higher than the original milk,

612

respectively.

EP

TE D

M AN U

SC

RI PT

597

Kaniou-Grigoriadou et al. (2005) reached a main enrichment factor of 4.9-fold for AFM1 in

614

curd produced with ewe’s milk and no AFM1 was found in Feta cheese after a 2-month ripening

615

period. Motawee and McMahon (2009) also produced feta cheese with artificially contaminated

616

milk (1 and 2 µg/kg AFM1) and observed a 64.1% retention of AFM1 in the cheese, which

617

represented a 3-fold concentration of the toxin. Statistical analysis did not show any significant

618

difference (P>0.05) in the partitioning of AFM1 between curd and whey as compared to the initial

619

AFM1 level in milk. These differences may be originated from the cheese making process,

620

chemical compositions of the cheeses, analytical methods, type and degree of milk contamination

AC C

613

ACCEPTED MANUSCRIPT 27 (Blanco et al., 1988; Oruc et al., 2006). Brackett et al. (1982) produced brick cheese and stated

622

that AFM1 concentration in the curd was 1.7-fold higher compared to the milk (12% of total toxin

623

in milk), while the whey contained about the same concentration than that found in milk. This

624

may occur because of the low yield of the curd and the small curd particles contained in the

625

whey. Oruc et al. (2007) showed that the concentrations of AFM1 in the curd were 2.9, 3.2 and

626

3.4-fold higher compared to that in the milk (50, 250 and 750 ng/L AFM1, respectively). The

627

distribution of AFM1 was 40-46% in the curd and 53-58% in the whey, and by increasing the

628

initial contamination level, the higher percentage of AFM1 was found in the curd and,

629

subsequently, in the cheese.

M AN U

SC

RI PT

621

The distribution of AFM1 in the curd and in the whey of Manchego cheese (a traditional

631

Spanish whey cheese) was studied by Rubio et al. (2011). They used artificially contaminated

632

raw ewe’s milk and the mean concentrations of AFM1 in the curd and in the final cheese were

633

about 2 and 3-fold higher than the level found in milk, respectively. The levels of remained toxin

634

in whey were 42.3% and 51.3% of the initial level in the curd and in the cheese. In Requesón

635

cheese, the mean AFM1 level was 1.7-fold higher than the related whey, while 33.7% to 44.4% of

636

the toxin concentration detected in the milk was found in the Requesón whey. Kamkar et al.

637

(2008) investigated the fate of AFM1 in Iranian Cheese processing using milk samples artificially

638

contaminated by AFM1 (0.25, 0.5, 0.75, 1, 1.25 and 1.75 µg/L). The mean AFM1 levels in the

639

curd and in the cheese were 3.12 and 3.65-fold more than the level found in the whey, and 1.68-

640

1.80-fold more than the level in cheese milk, respectively.

AC C

EP

TE D

630

641

Some researchers explain that AFM1 concentrations are 3 to 16-fold higher in the cheese

642

than in milk, while other studies have also reported it is approximately three times higher in soft

643

cheeses and five times higher in hard cheese as compared to the milk (Ardic et al., 2009).

644

Affinity of AFM1 to casein could be mentioned as a reason for increasing of AFM1 in cheese.

ACCEPTED MANUSCRIPT 28 Brackett and Marth (1982a) studied the binding process of AFM1 to casein using equilibrium

646

dialysis. They observed that casein suspensions had 2.5 and 2.9-fold more toxin when dialyzed

647

with 10 or 20 ng/mL of AFM1, respectively. Also, 30.7% more AFM1 was found when the casein

648

solutions were treated with a proteolytic enzyme, showing that AFM1 was bound to the casein

649

micelles, but not indicating whether the toxin was released or if the extraction procedure was

650

more efficient. In addition, it is possible to extract AFM1 from cheese due to its affinity for

651

casein, and that it may not be covalently linked, but linked to hydrophobic areas of casein by

652

hydrophobic interactions (Applebaum and Marth, 1982a).

SC

RI PT

645

Barbiroli et al. (2007) studied the affinity of AFM1 for casein in goat and sheep milk using

654

different methods: ultra-filtration, acid and enzymatic coagulation, and ricotta production (by

655

thermo-acid coagulation of the whey). Although ultra-filtration is used for selective separation of

656

certain toxic constituents of milk, it was not a suitable method for removing AFM1 from milk as

657

80% of toxin was retained in the protein-containing fraction (permeate), as the pores were not

658

sufficiently small to retain the AFM1. None of the treatments caused a significant decrease in

659

AFM1 level, despite the significant changes in the interaction with proteins. According to the

660

authors, the combination of acid and heat treatment used in the ricotta production was the only

661

process able to change the whey protein structures, allowing them to lose the ability to bind the

662

mycotoxin. Mendonça and Venâncio (2005) showed that the levels of AFM1 in permeate

663

(lactose-rich fraction of whey) was lower than in retentate (protein-rich fraction of whey) after

664

ultra-filtration. The difference can be explained by the affinity of AFM1 to the protein fraction of

665

the whey. As a result, the WPC (Whey protein powder concentrate) contained higher levels than

666

the whey. Therefore, the AFM1 level in WPC can be considered hazardous from a public health

667

standpoint.

668

AC C

EP

TE D

M AN U

653

ACCEPTED MANUSCRIPT 29 669

4.2) Effect of salting The salt content of cheese diverges markedly according to its type, ranging from 0.5 to

671

0.7% (w/w) in acid curd cheeses to about 4 to 6% (w/w) in pickled cheeses. The main functions

672

of salt in cheese are related to the cheese conservation and flavor enhancement. There are three

673

different types of salting: dry salting (direct addition of salt in the curd before molding), surface

674

dry salting (rubbing of salt on the cheese surface) and brine salting/brining (immersion of cheese

675

in brine solution) (Guinee, 2004). There are few existing studies considering the evaluation of

676

AFM1 content in cheeses after salting, and all of them used the brining method. Motawee and

677

McMahon (2009) stored Feta cheese in brining solutions of 8%, 10% and 12% (w/w) brine either

678

at 6oC or 18oC during 60 days and observed that the AFM1 level was significantly reduced during

679

the first 10 days, but no differences were observed after the 10th day.. Authors explained that the

680

reduction of AFM1 during the salting process of cheese is a function of the water solubility of

681

AFM1 and its diffusion into the brine, salt concentration and temperature can influence the effects

682

on expulsion/absorption of water by the cheese. Oruc et al. (2006) found that only 2–4% of the

683

initial spiking of AFM1 transferred into the brine solution after preparation of traditional Turkish

684

white-pickled cheese. The concentration of AFM1 in brine was low at the beginning and

685

increased towards the end of the ripening period (from 6 to 39 ng/L of AFM1 for the cheese

686

produced with milk contaminated with 50 ng/L AFM1, from 40 to 169 ng/L for milk with 250

687

ng/L AFM1 and from 110 to 344 ng/L for milk contaning 750 ng/L AFM1). While an increase of

688

AFM1 content in brine occurred, the toxin in cheese decreased. However, AFM1 levels in brine at

689

the end of the ripening were lower than the levels found in the curd/whey. Govaris et al. (2001)

690

also observed an increase in AFM1 concentration in brine at the end of the storage period (from

691

0.001 to 0.025 µg/kg AFM1 for the cheese produced with milk containing 0.05 µg/L AFM1 and

692

from 0.003 to 0.065 µg/kg for cheese with 0.10 µg/L AFM1 in the milk). The authors

AC C

EP

TE D

M AN U

SC

RI PT

670

ACCEPTED MANUSCRIPT 30 693

complemented that only a portion of the amount of AFM1 lost from cheese was found in brine,

694

which implies that the remaining amount of AFM1 lost from cheese might be degraded during

695

ripening.

697

RI PT

696 4.3) Effect of ripening process

Ripening is an important step in cheese making process because during this period

699

physicochemical reactions and microbiological modifications invariably occur, developing

700

volatile organic compounds, texture and flavor in the cheese stored under controlled temperature

701

and humidity. The fate of AFM1 in cheese ripening has been evaluated: Brackett and Marth

702

(1982b) produced cheddar cheese from AFM1 naturally contaminated milk and observed that

703

AFM1 content during the ripening period started low (6-11 µg/kg), than rose to their highest

704

values (12-23 µg/kg) at about 18-24 weeks and dropped to their initial values (3-14 µg/kg) at the

705

end of maturation (40 weeks). During Parmesan cheese ripening time (43 weeks), AFM1 content

706

started high (35-66 µg/kg), decreased until the 22nd week (4-7 µg/kg) and increased to a final

707

concentration of 24-30 µg/kg. Authors suggested that the action of enzymes (lipase) somehow

708

affects the behavior of the toxin in cheese and allows a more efficient recovery of AFM1 during

709

analysis (Brackett and Marth, 1982c). During a three-month ripening period of traditional white-

710

pickled cheese, no significant degradation of AFM1 was observed (Oruc et al., 2006).

AC C

EP

TE D

M AN U

SC

698

711

Manetta et al. (2009) observed a 4.5-fold higher content of AFM1 in Grana Padano cheese

712

compared to the original milk after 20 months of ripening. The authors obtained a positive

713

correlation between milk and cheese showing that AFM1 concentration in milk is a good

714

predictor of its fate in Grana Padano cheese and it can help the producers to estimate the expected

715

AFM1 level in cheese made from contaminated milk. Govaris et al. (2001) showed that AFM1

716

concentration in Telemes cheese did not remain constant during a ripening/storage period of 6

ACCEPTED MANUSCRIPT 31 months. Significant decrease (P<0.05) in the concentrations of AFM1 was observed after 60 days.

718

Another significant decrease occurred at day 120 in the cheese with a higher dose (0.10 µg/L),

719

while the cheese with a lower dose did not present differences. Reduction of AFM1 was higher at

720

the end of the second month of ripening as compared to the level found at the end of the fourth or

721

the sixth month of storage. Brackett et al. (1982) produced brick cheese and quantified the AFM1

722

level during 26 weeks. There was a difference in the AFM1 content at different locations in the

723

cheeses during the ripening period in a way that cheeses ripened with a smear for 2 or 3 weeks

724

presented a higher AFM1 level in the rind, while cheeses with a surface smeared for 4 weeks

725

presented a high AFM1 level in center. These differences may be derived from the inward

726

diffusion of enzymes involved in surface-ripening or from a faster ripening of the cheese on the

727

surface without diffusion of enzymes. Conversely, Blanco et al. (1988) found no differences in

728

aflatoxin concentration between inner and outer portions of cheese after 60 days. The authors

729

resumed that the causing factors are: effect of cheese microorganisms; difficulty in the aflatoxin

730

extraction from cheese, effect of different ripening products and cheese size; chemical or

731

enzymatic degradation processes that occur during cheese ripening; type of cheese manufacture

732

operations; peptides produced during proteolysis that could be responsible for the partial

733

destruction of aflatoxins.

735

SC

M AN U

TE D

EP

AC C

734

RI PT

717

4.4) Effect of storage

736

Cheese storage can be performed at room or refrigerated temperature, depending on the

737

type of cheese. Fremy et al. (1990) studied the influence of storage in Camembert cheeses

738

produced with raw milk spiked with 7.5, 3.0 and 0.3 µg/L AFM1 and observed that in the first 15

739

days of storage, AFM1 level decreased 25%, 55% and 75%, respectively. AFM1 content in

740

Mozarella cheese during storage (7oC, 19 weeks) remained relatively constant. According to the

ACCEPTED MANUSCRIPT 32 authors, it is possible that some steps used to produce this cheese (direct acidification and heating

742

in water at 80oC before stretching and molding of the curd) allow maximum AFM1 levels to be

743

recovered during storage (Brackett and Marth, 1982c). Oruc et al. (2007) concluded that AFM1

744

was relatively stable (2-4%) in Kashar cheese during a 2-month storage period. Blanco et al.

745

(1988) studied the frozen storage of Manchego-type cheese during 90 days and observed only

746

slight variations (<20%) in aflatoxin content. Studies regarding the AFM1 stability during cheese

747

ripening and storage are not consensual. As well described by Fernandes et al. (2012), the

748

stability of AFM1 during storage depends on the type of cheese. According to the authors, after

749

30 days of storage of Minas Frescal cheese, AFM1 level was 2.14 to 2.60-fold higher than the

750

levels found in milk. Some cheeses, including Brick, Limburger, Camembert, Tilsit, Cheddar,

751

Gouda, Manchego, Parmesan, Mozzarella and Swiss show a considerable stability of AFM1

752

during maturation and storage (Galvano et al., 1996; Kamkar et al., 2008). It is important to keep

753

cheese and fermented dairy products under refrigerated storage, combined with good sanitation

754

and handling practices throughout the food chain, and whenever possible, it is advisable to

755

exclude air from the packaging by using vacuum, as these measures help to prevent and/or

756

minimize the growth of potentially toxic molds (Bullerman, 1981). As both milk pasteurization

757

and cheese manufacture processes do not eliminate AFM1, it is prudent to check the AFM1

758

incidence in cheese (Prado et al., 2001).

760 761

SC

M AN U

TE D

EP

AC C

759

RI PT

741

5) The fate of AFM1 during yoghurt processing The basic steps involved in the production of yoghurt are described in Figure 4.

762

Variations in the process can occur because of the type of yoghurt produced, that is, the

763

traditional (with a firmer consistency) or the stirred type. In this section, the fate of AFM1 in the

764

yoghurt manufacture operations will be considered.

ACCEPTED MANUSCRIPT 33 765 766

Insert Figure 4 here.

767 5.1) Effect of fermentation

RI PT

768

Studies about the stability of AFM1 in yoghurt during fermentation are limited and

770

controversial. Iha et al. (2013) reached a 6.4% decrease of AFM1 content after yoghurt processing

771

and Bakirci (2001) obtained a 13% higher AFM1 concentration than the level initially found in

772

milk. In the work of Şanli et al. (2012), AFM1 level decreased 36.5% and 34.6% compared to the

773

original milk respectively, in yoghurts produced with AFM1 artificially contaminated milk at 1.5

774

and 3.5 µg/kg. Govaris et al. (2002) produced yoghurt using AFM1 artificially contaminated milk

775

(0.05 and 0.10 µg/L) and observed that AFM1 levels in yoghurt reduced significantly by 13% and

776

22%, respectively, compared to the control. Lower AFM1 levels were found in yoghurts at pH 4.0

777

as compared to pH 4.6, and the authors attributed this result to the metabolic activity of LAB,

778

with great production of lactic acid and other byproducts. Changes in casein structure during

779

yoghurt making can affect the association of AFM1 with casein, causing adsorption or occlusion

780

of AFM1 in the precipitate. Jasutiene et al. (2006) observed that the fermentation process reduced

781

the AFM1 level by 22-28% compared to the initial level of different samples. Fermentation

782

conducted at pH 4.0 and pH 4.5 had no significant effect on the stability of the toxin. According

783

to the authors, the effect of low pH could cause decomposition of the milk structure and, as a

784

result, AFM1 could be associated with casein and it lead to a considerable AFM1degradation.

M AN U

TE D

EP

AC C

785

SC

769

The contradictory results on the stability of AFM1 during processing and storage are

786

because of different pHs of yoghurts, variable AFM1 levels in milk, different fermentation

787

conditions, changes in physicochemical properties of caseins and variations in analytical methods

788

(Govaris et al., 2002). According to Arab et al. (2012), the disadvantages of aflatoxin occurrence

ACCEPTED MANUSCRIPT 34 in fermented milks are: longer fermentation time, decrease in the growth, morphology, and

790

activity rate of starter cultures, defects in flavor and texture, conversion of homofermentative

791

starter cultures into heterofermentative ones. Nutritional aspects of yoghurt consumption is

792

associated with the presence and viability of microorganisms, but aflatoxins can change the

793

appearance or cause negative effects on nutritional properties of these products (Galvano et al.,

794

1996). Some adverse effects of AFM1 were observed in Lactobacillus bulgaricus, such as

795

increased thickness and shortened chain length of cells, whereas in Streptococcus thermophilus,

796

the wall became thicker and changes occurred in its shape (El Deeb et al., 1992). Govaris et al.

797

(2002) observed that populations of LAB were not affected in contaminated yoghurts, except for

798

S. thermophilus that presented a significant lower increase rate in the yoghurt with higher toxin

799

concentration (0.10 µg/L AFM1).

M AN U

SC

RI PT

789

As result of potential danger to human health, especially children, there are many

801

investigations to find suitable methods to remove or inactivate AFM1 in milk and dairy products.

802

LAB strains commonly used in the production of yoghurt and other dairy products can have a

803

role in the removal of AFM1. Some studies have shown that these bacteria have the ability to bind

804

the molecule of AFM1 reducing its absorption in the gastrointestinal tract and, consequently,

805

decreasing its toxicological effects. Pierides, El-Nezami et al. (2000) analysed 6 strains of LAB

806

(Lactobacillus acidophilus LA1, Lactobacillus gasseri ATCC 33323, L. rhamnosus GG and LC-

807

705, L. rhamnosus 1/3 and Lactococcus lactis ssp. cremoris ARH74) for their ability to remove

808

AFM1 from a phosphate buffer saline (PBS) solution. Binding results varied from 18.1% to

809

53.8%, and L. rhamnosus GG and LC-705 showed a better performance in removing AFM1

810

(50.7% and 46.3%, respectively). According to this report, the inactivation of LAB cells caused

811

by boiling enhanced the AFM1 removal (25.5% to 61.5%). When milk was used as the medium

812

for binding tests, the AFM1 removal of viable L. rhamnosus GG decreased in skim and full cream

AC C

EP

TE D

800

ACCEPTED MANUSCRIPT 35 milk (18.8% and 26.0%, respectively), while heat-killed L. rhamnosus GG removed 26.6% and

814

36.6%, respectively. Authors explained that the less effective removal in milk compared to PBS

815

may be because of AFM1 might not be accessible in milk as it may be linked to casein. In

816

addition, the interference of proteins in the removal process can be responsible for the differences

817

between the full cream and skim milk (approximately 10% smaller), as the skim milk powder

818

contained 37 g protein/100 g, while the full cream milk powder contained 25 g protein/100 g.

819

Kabak and Var (2008) observed that viable cells 4 strains of Lactobacillus and 2 strains of

820

bifidobacteria bound 10.2% to 26.7% of AFM1 in PBS while heat-killed cells bound 14.0% to

821

29.0% of AFM1. In reconstituted milk, binding porcentages were 7.9% to 26.0% and 12.9% to

822

27.3% for viable and heat-killed cells, respectively. Bovo et al. (2012) evaluated seven LAB

823

strains and observed that AFM1 removal ranged from 5.6% to 45.7% in PBS, and L. rhamnosus,

824

L. bulgaricus and Bifidobacterium lactis presented the best results. These strains, when analysed

825

in UHT skim milk showed AFM1 binding capacity of 13.5% to 37.8%. Corassin et al. (2013)

826

tested the ability of a mixture of S. cerevisiae and three LAB strains (L. rhamnosus, L. bulgaricus

827

and B. lactis), alone or in combination, to bind AFM1 in UHT skim milk. According to the

828

results, a mean AFM1 binding caused by the LAB pool in milk was 11.5%, while S. cerevisiae

829

cells bound 90% of AFM1. In addition, S. cerevisiae + LAB pool bound 100% of AFM1.

830

Controversial results in the AFM1 binding can be accounted by differences in the toxin

831

concentration, viability of cells, extraction procedures, contact time between the toxin and the

832

cells, temperature, composition of the medium (especially protein and fat contents) and the

833

chosen strains, as not all strains have similar aflatoxin binding properties.

AC C

EP

TE D

M AN U

SC

RI PT

813

834

Some studies analysed yoghurt starter cultures. Sarimehmetoğlu and Küplülü (2004)

835

studied the removal activities of AFM1 by L. bulgaricus CH-2 and S. thermophilus ST-36 in PBS,

836

contaminated reconstituted milk and contaminated yoghurt made from reconstituted milk. The

ACCEPTED MANUSCRIPT 36 837

result showed that in PBS and in milk, L. bulgaricus CH-2 bound 18.7% and 27.6% of AFM1,

838

respectively, while S.

839

Complementarialy, 14.8% AFM1 was bound in the yoghurt matrix. Elsanhoty et al. (2014)

840

evaluated the ability of six LAB strains (Lactobacillus acidophilus ATCC 20552, L. rhamnosus

841

TISTR 541, Lactobacillus plantrium, Bifidobacterium angulatum DSMZ 20098, S. thermophilus

842

and L. bulgaricus) for their AFM1 binding capacity. In PBS, AFM1 binding varied from 63.2% to

843

79.2% for viable cells after an incubation period of 4 h, and from 86.8% to 95.8% for heat-treated

844

cells. Yoghurt produced by S. thermophilus and L. bulgaricus had 69.8% less AFM1 after 7 days

845

of storage, while the yoghurt produced with S. thermophilus, L. bulgaricus and L. plantrium

846

presented 87.8% less AFM1. Likewise, the yoghurt produced with S. thermophilus, L. bulgaricus

847

and L.acidophilus had 72.8% less AFM1. The effect of L. bulgaricus and S. thermophilus used in

848

the Lebanese traditional industry was investigated for their ability to reduce the level of AFM1 by

849

El Khoury et al. (2011). The results showed that L. bulgaricus had the highest AFM1 binding

850

ability (87.6% after 14 h) compared to S. thermophilus (70% after 14 h). When the cultures were

851

combined, a lower binding capacity (26.1% within 2 h and 68% within 14 h) was obtained,

852

confirming the effect of time (kinetics) on removing the toxin.

39.2%, respectively.

TE D

M AN U

SC

RI PT

bound 29.2% and

EP

853

thermophilus ST-36

855

AC C

854 5.2) Effect of cold storage

Again, results obtained for AFM1 fate during storage of yoghurt are controversial, besides

856 there are few available. Wiseman and Marth (1983) observed that AFM1 remained in yoghurt 857 during a storage period of 6 weeks, and AFM1 content fluctuated with the slow decrease of yoghurt 858 pH. According to the authors, changes in pH can affect the protein structure and influence the 859 capability of chloroform to extract AFM1 from the casein net, and this may explain the differences 860 found during this period. Hassanin (1994) indicated that after 14 days of storage, only 41% of

ACCEPTED MANUSCRIPT 37 861 initial level of AFM1 could be recovered from yoghurt. Nevertheless, it should be mentioned that 862 reduced recovery does not mean a reduction in toxicity of AFM1. Iha et al. (2013) obtained a 863 reduction of only 6% for yoghurt (pH 4.4) during a 4-week storage period. During storage (4oC/4

RI PT

864 weeks), AFM1 was more stable in yoghurt at pH 4.6 than in yoghurt at pH 4.0, reducing 16% and 865 34%, respectively (Govaris et al., 2002). Şanli et al. (2012) observed that the AFM1 level was 866 reduced by 6.5% after a 2-week storage period compared to the yoghurt produced with AFM1

SC

867 artificially contaminated milk at 1.5 and 3.5 µg/kg. Recently, results of Aly and Diekmanns (2010) 868 showed that the toxin level in yoghurt did not significantly change during the storage period.

M AN U

869 According to the authors, in some cases, the decrease of AFM1 during the storage of yoghurt in 870 refrigerated conditions can be a result of dissociation of milk proteins (casein). 871 872

6) Conclusions and final remarks

Data of the present study demonstrated that literature shows variable findings regarding

874

AFM1 reduction during different unit operations used in dairy products processing. In addition,

875

the current legislation worldwide is not compatible, as each country has its own regulation, which

876

hinders the commercialization of products in the international market, in addition to the

877

inexistence of legislation for all dairy products produced/consumed worldwide.

EP

TE D

873

The effects associated with the aflatoxin intake are very severe and can trigger a varied

879

number of health implications, especially if long-term effects are considered. It is extremely

880

important to control the quality of the animal feed for the lactating animals in order to avoid the

881

transformation of AFB1 into AFM1, in addition to adopting preventive measures to avoid optimal

882

growth conditions and mycotoxin production. Therefore, further studies concerning the

883

occurrence and stability of aflatoxin in dairy products should be carried out in order to avoid the

884

toxicological effects in humans and animals.

AC C

878

ACCEPTED MANUSCRIPT 38 885 886

Acknowledgements The authors acknowledge the financial support of “Fundação de Amparo a Pesquisa do

888

Estado de São Paulo” (FAPESP) (Grant #14/14891-7), “Conselho Nacional de Desenvolvimento

889

Científico e Tecnológico” (CNPq) (Grant #302763/2014-7), CNPq-TWAS Postgraduate

890

Fellowship (Grant #3240274290), and Coordenação de Aperfeiçoamento de Pessoal de Nível

891

Superior (CAPES) for the financial support.

SC

RI PT

887

892 Conflict of interest

894

The authors declare no conflict of interest.

M AN U

893

895 7) References

897

Abrar, M., Anjum, F. M., Butt, M. S., Pasha, I., Randhawa, M. A., Saeed, F., & Waqas, K.

TE D

896

898

(2013). Aflatoxins: biosynthesis, occurrence, toxicity and remedies. Critical Reviews in

899

Food Science, 53, 862-874.

901

Alborzi, S., Pourabbas, B., Rashidi, M., & Astaneh, B. (2006). Aflatoxin M1 contamination in

EP

900

pasteurized milk in Shiraz (south of Iran). Food Control, 17, 582–584. Ali, M. A. I., El Zubeir, I. E. M., & Fadel Elseed, A. (2014). Aflatoxin M1 in raw and imported

903

powdered milk sold in Khartoum state, Sudan. Food Additives and Contaminants - Part B,

904

7, 208–212.

905 906

AC C

902

Aly, S.A., & Diekmanns, H. (2010). Fate of Aflatoxin M1 during manufacture and storage of yoghurt. Milchwissenschaft, 65, 193-194.

907

Anfossi, L., Baggiani, C., Giovannoli, C., & Giraudi, G. (2011). Occurrence of Aflatoxin M1 in

908

Dairy Products. In I. Torres-Pacheco (Ed.), Aflatoxins – Detection, Measurement and

ACCEPTED MANUSCRIPT 39 909

Control (pp. 3-20). INTECH Open Access Publisher.

910

http://cdn.intechweb.org/pdfs/22029.pdf. Accessed 10.10.15.

911

Anfossi, L., Baggiani, C., Giovannoli, C., D’Arco, G., Passini, C., & Giraudi, G. (2012). Occurrence of aflatoxin M1 in Italian cheese: Results of a survey conducted in 2010 and

913

correlation with manufacturing, production season, milking animals, and maturation of

914

cheese. Food Control, 25, 125–130.

ANVISA – Agência Nacional de Vigilância Sanitária, Brazil. Resolução RDC n°7, de 18 de

SC

915

RI PT

912

fevereirode 2011. Regulamento Técnico sobre limites máximos tolerados (LMT) para

917

micotoxinas em alimentos. (2011).

918

http://portal.anvisa.gov.br/wps/wcm/connect/bc17db804f45fe2cbd41fdd785749fbd/Resolu

919

%C3%A7%C3%A3o+0-2011-GGALI.pdf?MOD=AJPERES. Accessed 25.10.15.

921

Applebaum, R.S., & Marth, E. (1982a). Fate of aflatoxin M1 in cottage cheese. Journal of Food Protection, 45, 903–904.

TE D

920

M AN U

916

Applebaum, R. S., & Marth, E. H. (1982b). Use of sulphite or bentonite to eliminate aflatoxin M1

923

from naturally contaminated raw whole milk. Zeitschrift Für Lebensmittel-Untersuchung

924

Und -Forschung, 174, 303-305.

925

EP

922

Arab, M., Sohrabvandi, S., Mortazavian, A. M., Mohammadi, R., & Tavirani, M. R. (2012). Reduction of aflatoxin in fermented milks during production and storage. Toxin Reviews, 31,

927

44–53.

928 929 930

AC C

926

Ardic, M., Karakaya, Y., Atasever, M., & Adiguzel, G. (2009). Aflatoxin M1 levels of Turkish white brined cheese. Food Control, 20, 196–199. Asi, M. R., Iqbal, S. Z., Ariño, A., & Hussain, A. (2012). Effect of seasonal variations and

931

lactation times on aflatoxin M1 contamination in milk of different species from Punjab,

932

Pakistan. Food Control, 25, 34-38.

ACCEPTED MANUSCRIPT 40 933 934 935

Aycicek, H., Aksoy, A., & Saygi, S. (2005). Determination of aflatoxin levels in some dairy and food products which consumed in Ankara, Turkey. Food Control, 16, 263–266. Awasthi, V., Bahman, S., Thakur, L. K., Singh, S. K., Dua, A., & Ganguly, S. (2012). Contaminants in milk and impact of heating: an assessment study. Indian Journal of Public

937

Health, 56, 95–9.

939 940

Bakirci, I. (2001). A study on the occurrence of aflatoxin M1 in milk and milk products produced in Van province of Turkey. Food Control, 12, 47–51.

SC

938

RI PT

936

Baptista, A. S., Horii, J., & Baptista, A. S. (2004). Fatores físico-químicos e biológicos ligados à produção de micotoxinas. Boletim do Centro de Pesquisa e Processamento de Alimentos,

942

22, 1-14.

944 945 946 947

Baranyi, N., Kocsubé, S., Vágvölgyi, C., & Varga, J. (2013). Current trends in aflatoxin research. Acta Biologica Szegediensis, 57, 95-107.

Baranyi, N., Kocsubé, S., & Varga, J. (2015). Aflatoxins: Climate change and biodegradation.

TE D

943

M AN U

941

Current Opinion in Food Science, 5, 60–66. Barbiroli, A., Bonomi, F., Benedetti, S., Mannino, S., Monti, L., Cattaneo, T., & Iametti, S. (2007). Binding of aflatoxin M1 to different protein fractions in ovine and caprine milk.

949

Journal of Dairy Science, 90, 532–40. Battacone, G., Nudda, A., Cannas, A., Cappio Borlino, A., Bomboi, G., & Pulina, G. (2003).

AC C

950

EP

948

951

Excretion of aflatoxin M1 in milk of dairy ewes treated with different doses of aflatoxin B1.

952

Journal of Dairy Science, 86, 2667–2675.

953

Battacone, G., Nudda, A., Palomba, M., Pascale, M., Nicolussi, P., & Pulina, G. (2005). Transfer

954

of Aflatoxin B1 from Feed to Milk and from Milk to Curd and Whey in Dairy Sheep Fed

955

Artificially Contaminated Concentrates. Journal of Dairy Science, 88, 3063-3069.

ACCEPTED MANUSCRIPT 41 956

Bartoszek, A. (2006). Genotoxic Food Components. In D. Malejka-Giganti, A. Bartoszek, & W.

957

Baer-Dubowska (Eds.), Carcinogenic and Anticarcinogenic Food Components (pp. 69-96).

958

Boca Raton: CRC Taylor & Francis Group.

960

Beltrane, M. A., & Machinski, M. J. (2005). Principais Riscos Químicos no Leite: um problema

RI PT

959

de Saúde Pública. Arquivos de Ciências da Saúde da UNIPAR, 9,141-145.

Bhat, R., Rai, V. R., & Karim, A. A. (2010). Mycotoxins in Food and Feed. Present status and

962

future concerns. Comprehensive Reviews in Food Science and Food Safety, 9, 57–81.

964 965

Bilandžić, N., Varenina, I., & Solomun, B. (2010). Aflatoxin M1 in raw milk in Croatia. Food Control, 21, 1279–1281.

M AN U

963

SC

961

Bilandžić, N., Božić, Đ., Đokić, M., Sedak, M., Kolanović, B. S., Varenina, I., & Cvetnić, Ž.

966

(2014a). Assessment of aflatoxin M1 contamination in the milk of four dairy species in

967

Croatia. Food Control, 43, 18–21.

Bilandžić, N., Božić, D., Dokić, M., Sedak, M., Kolanović, B. S., Varenina, I., Tanković, S., &

969

Cvetnić, Ž. (2014b). Seasonal effect on aflatoxin M1 contamination in raw and UHT milk

970

from Croatia. Food Control, 40, 260–264.

Blanco, J. L., Dominguez, L., Gomez-Lucia, E., Garayzabal, J. F. F., Garcia, J. A., & Suarez, G.

EP

971

TE D

968

(1988). Presence of Aflatoxin M1 in Commercial Ultra-High-Temperature-Treated Milk.

973

Applied and Environmental Microbiology, 54, 1622–1623.

974

AC C

972

Bovo, F., Corassin, C. H., Rosim, R. E., & Oliveira, C. A. F. (2012). Effiency of lactic acid

975

bacteria strains for decontamination of aflatoxin M1 in phosphate buffer saline solution and

976

in skim milk. Food and Bioprocess Technology, 5, 1-5.

977

Bovo, F., Franco, L. T., Rosim, R. E., Barbalho, R., & Oliveira, C. A. F. (2015). In vitro ability

978

of beer fermentation residue and yeast-based products to bind aflatoxin B1. Brazilian

979

Journal of Microbiology, 46, 577-581.

ACCEPTED MANUSCRIPT 42

983 984 985 986 987 988 989 990 991

Brackett, R. E., & Marth, E. H. (1982a). Association of aflatoxin M1 with casein. Zeitschrift für Lebensmittel-Untersuchung und -Forschung, 174, 439–441.

RI PT

982

in brick and Limburger-like cheese. Journal of Food Protection, 45, 553-556.

Brackett, R. E., & Marth, E. H. (1982b). Fate of Aflatoxin M1 in Cheddar cheese and in process spread. Journal of Food Protection, 45, 549–552.

Brackett, R. E, & Marth, E. H. (1982c). Fate of Aflatoxin M1 in Parmesan and Mozzarella cheese. Journal of Food Protection, 45, 597–600.

SC

981

Brackett, R. E., Applebaum, R. A., Wiseman, D. W., & Marth, E. H. (1982). Fate of aflatoxin M1

Bryden, W. L. (2012). Mycotoxin contamination of the feed supply chain: Implications for

M AN U

980

animal productivity and feed security. Animal Feed Science and Technology, 173, 134-158. Bullerman, L. B. (1981). Public Health Significance of Molds and Mycotoxins in Fermented Dairy Products. Journal of Dairy Science, 64, 2439–2452.

Cano-Sancho, G., Marin, S., Ramos, A. J., Peris-Vicente, J., & Sanchis, V. (2010). Occurrence of

993

aflatoxin M₁ and exposure assessment in Catalonia (Spain). Revista Iberoamericana de

994

Micología, 27, 130–135.

Carraro, A., De Giacomo, A., Giannossi, M. L., Medici, L., Muscarella, M., Palazzo, L.,

EP

995

TE D

992

Quarantab, V., Summac, V., Tateo, F. (2014). Clay minerals as adsorbents of aflatoxin M1

997

from contaminated milk and effects on milk quality. Applied Clay Science, 88-89, 92-99.

998 999 1000

AC C

996

Castegnaro, M., & Mcgregor, D. (1998) Carcinogenic risk assessment of mycotoxins. Revue de Medecine Veterinaire, 149, 671–678. Cavallarin, L., Antoniazzi, S., Giaccone, D., Tabacco, E., & Borreani, G. (2014). Transfer of

1001

aflatoxin M1 from milk to ripened cheese in three Italian traditional production methods.

1002

Food Control, 38, 174–177.

ACCEPTED MANUSCRIPT 43 1003 1004

Çelik, T. H., Sarımehmetoğlu, B., & Küplülü, Ö. (2005). Aflatoxin M1 contamination in pasteurised milk. Veterinarski Arhiv, 75, 57–65. Chopra, R. C., Chabra, A., Prasad, K. S. N., Dudhe, A., Murthy, T. N., & Prasad, T. (1999).

1006

Carryover of aflatoxin M1 in milk of cows fed aflatoxin B1 contaminated ration. Indian

1007

Journal of Animal Nutrition, 16, 103-106

1009

Creppy, E. E. (2002). Update of survey, regulation and toxic effects of mycotoxins in Europe. Toxicology Letters, 127, 19-28.

SC

1008

RI PT

1005

Codex Alimentarius Commission. Comments submitted on the draft maximum level for aflatoxin

1011

M1 in milk. Codex Committee on food additives and contaminants 33rd session, Hague, The

1012

Netherlands. (2001). ftp://ftp.fao.org/codex/Meetings/CCFAC/ccfac33/fa01_20e.pdf.

1013

Accessed 15.10.15.

1014

M AN U

1010

Colak, H. (2007). Determination of Aflatoxin M1 Levels in Turkish White and Kashar Cheeses Made of Experimentally Contaminated Raw Milk. Journal of Food and Drug Analysis, 15,

1016

163–168.

1017

TE D

1015

Corassin, C. H., Bovo, F., Rosim, R. E., & Oliveira, C. A. F. (2013). Efficiency of Saccharomyces cerevisiae and lactic acid bacteria strains to bind aflatoxin M1 in UHT skim

1019

milk. Food Control, 31, 80–83.

EP

1018

Dashti, B., Al-Hamli, S., Alomirah, H., Al-Zenki, S., Abbas, A. B., & Sawaya, W. (2009). Levels

1021

of aflatoxin M1 in milk, cheese consumed in Kuwait and occurrence of total aflatoxin in

1022

local and imported animal feed. Food Control, 20, 686–690.

1023

AC C

1020

Decastelli, L., Lai, J., Gramaglia, M., Monaco, A., Nachtmann, C., Oldano, F., Ruffier, M.,

1024

Sezian, A., & Bandirola, C. (2007). Aflatoxins occurrence in milk and feed in Northern Italy

1025

during 2004-2005. Food Control, 18, 1263–1266.

ACCEPTED MANUSCRIPT 44

1028 1029 1030 1031 1032

and storage of skim milk powder. Journal of Food Protection, 69, 682-685. Deveci, O. (2007). Changes in the concentration of aflatoxin M1 during manufacture and storage of White Pickled cheese. Food Control, 18, 1103–1107.

RI PT

1027

Deveci, O, & Sezgin, E. (2006). Changes in concentration of aflatoxin M1 during manufacture

Di Natale, F., Gallo, M., & Nigro, R. (2009). Adsorbents selection for aflatoxins removal in bovine milks. Journal of Food Engineering, 95, 186-191.

Duarte, S. C., Almeida, A. M., Teixeira, A. S., Pereira, A. L., Falcão, A. C., Pena, A., & Lino, C.

SC

1026

M. (2013). Aflatoxin M1 in marketed milk in Portugal: Assessment of human and animal

1034

exposure. Food Control, 30, 411–417.

1035 1036 1037

M AN U

1033

Dutton, M. F., Mwanza, M., De Kock, S., & Khilosia, L. D. (2012). Mycotoxins in South African foods: A case study on aflatoxin M1 in milk. Mycotoxin Research, 28, 17–23. El-Deeb, S. A., Kheadr, E. E., Nadia, Z., & Shoukry Y. M. R. (1992). Effect of some technological processes on stability and distribution of Aflatoxin M1 in milk. Journal of

1039

Food Science, 20, 29-42.

1040

TE D

1038

El Khoury, A., Atoui, A., & Yaghi, J. (2011). Analysis of aflatoxin M1 in milk and yogurt and AFM1 reduction by lactic acid bacteria used in Lebanese industry. Food Control, 22, 1695–

1042

1699.

EP

1041

El-Nezami, H., Kankaanpaa, P., Salminem, S., & Ahokas, J. (1998). Ability of dairy strains of

1044

lactic acid bacteria to bind a common food carcinogen, Aflatoxin B1. Food and Chemical

1045

Toxicology, 36, 321-326.

AC C

1043

1046

El-Nezami, H., Mykkänen, H., Haskard, C., Salminen, S., & Salminen, E. (2004). Lactic Acid

1047

Bacteria as a Tool for Enhancing Food Safety by Removal of Dietary Toxins. In S.

1048

Salminen, A. Von Wright, & A. Ouwehand (Eds.), Lactic acid bacteria: microbiological

1049

and functional aspects (pp. 397-406). New York: Marcel Dekker.

ACCEPTED MANUSCRIPT 45 1050

Elgerbi, A. M., Aidoo, K. E., Candlish, A. A. G., & Tester, R. F. (2004). Occurrence of aflatoxin

1051

M1 in randomly selected North African milk and cheese samples. Food Additives and

1052

Contaminants, 21, 592–597.

1054 1055

Elkak, A., El Atat, O., Habib, J., & Abbas, M. (2012). Occurrence of aflatoxin M1 in cheese

RI PT

1053

processed and marketed in Lebanon. Food Control, 25, 140–143.

Elsanhoty, R. M., Salam, S. A., Ramadan, M. F., & Badr, F. H. (2014). Detoxification of

aflatoxin M1 in yoghurt using probiotics and lactic acid bacteria. Food Control, 43, 129–

1057

134.

El Marnissi, B., Belkhou, R., Morgavi, D. P., Bennani, L., & Boudra, H. (2012). Occurrence of

M AN U

1058

SC

1056

1059

Aflatoxin M1 in raw milk collected from traditional dairies in Morocco. Food and Chemical

1060

Toxicology, 50, 2819-2821.

Er, B., Demirhan, B., Onurdag, F. K., & Yentur, G. (2010). Determination of aflatoxin M1 level

1062

in milk and white cheese consumed in Ankara Region, Turkey. Journal of Animal and

1063

Veterinary Advances, 9, 1780-1784.

1064

TE D

1061

Ertas, N., Gonulalan, Z., Yildirim, Y., & Karadal, F. (2011). A survey of concentration of aflatoxin M1 in dairy products marketed in Turkey. Food Control, 22, 1956–1959.

1066

European Commission. Regulation (EC) No. 466/2001, setting maximum levels for certain

EP

1065

contaminants in foodstuffs. Official J. Eur. Communities L77, 1–13. (2001).

1068

http://ec.europa.eu/food/fs/sfp/fcr/fcr02_en.pdf. Accessed 15.10.15.

1069

AC C

1067

European Commission. Regulation. (EC) No. 1881. setting maximum levels for certain

1070

contaminants in foodstuffs (Text with EEA relevance). Official J. Eur. Communities L364,

1071

5–24. (2006). https://www.fsai.ie/uploadedFiles/Consol_Reg1881_2006.pdf. Accessed

1072

15.10.15.

ACCEPTED MANUSCRIPT 46 1073

Fallah, A. A., Jafari, T., Fallah, A., & Rahnama, M. (2009). Determination of aflatoxin M1 levels

1074

in Iranian white and cream cheese. Food and Chemical Toxicology, 47, 1872–1875.

1075

Fallah, A. A. (2010a). Assessment of aflatoxin M1 contamination in pasteurized and UHT milk

1078 1079

RI PT

1077

marketed in central part of Iran. Food and Chemical Toxicology, 48, 988–991.

Fallah, A. A. (2010b). Aflatoxin M1 contamination in dairy products marketed in Iran during winter and summer. Food Control, 21, 1478–1481.

Fallah, A. A., Rahnama, M., Jafari, T., & Saei-Dehkordi, S. S. (2011). Seasonal variation of

SC

1076

aflatoxin M1 contamination in industrial and traditional Iranian dairy products. Food

1081

Control, 22, 1653–1656.

1082

M AN U

1080

Fernandes, A. M., Corrêa, B., Rosim, R. E., Kobashigawa, E., & Oliveira, C. A. F. (2012).

1083

Distribution and stability of aflatoxin M1 during processing and storage of Minas Frescal

1084

cheese. Food Control, 24, 104–108.

Ferreira, H., Pittner, E., Sanches, H. F., & Monteiro, M. C. (2006). Aflatoxinas: um risco a saúde

1086

humana e animal. Ambiência - Revista do Centro de Ciências Agrárias e Ambientais, 2,

1087

113-127.

1090

EP

1089

Fink-Gremmels, J. (2008). Mycotoxins in cattle feeds and carry-over to dairy milk: a review. Food Additives and Contaminants, 25, 172-180. Fremy, J. M., Roiland, J. C., & Gaymard, D. (1990). Behavior of 14C aflatoxin M1 during

AC C

1088

TE D

1085

1091

Camembert cheese making. Journal of Environmental Pathology, Toxicology and Oncology,

1092

10, 95-98.

1093

Gallo, A., Masoero, F., Bertuzzi, T., Piva, G., & Pietri, A. (2010). Effect of the inclusion of

1094

adsorbents on aflatoxin B-1 quantification in animal feedstuffs. Food Additives and

1095

Contaminants - Part A Chemistry, Analysis, Control, Exposure and Risk Assessment, 27, 54-

1096

63.

ACCEPTED MANUSCRIPT 47 1097 1098 1099

Galvano, F., Galofaro, V., & Galvano, G. (1996). Occurrence and stability of aflatoxin M1 in milk and milk products. A worldwide review. Journal of Food Protection, 59, 1079-1090. Galvano, F., Galofaro, V., Ritieni, a, Bognanno, M., De Angelis, A., & Galvano, G. (2001). Survey of the occurrence of aflatoxin M1 in dairy products marketed in Italy: second year of

1101

observation. Food Additives and Contaminants, 18, 644–6.

1102

RI PT

1100

Garrido, N. S., Iha, M. H., Santos Ortolani, M. R., & Duarte Fávaro, R. M. (2003). Occurrence of aflatoxins M(1) and M(2) in milk commercialized in Ribeirão Preto-SP, Brazil. Food

1104

Additives and Contaminants, 20, 70–73.

1107 1108 1109

M AN U

1106

Ghanem, I., & Orfi, M. (2009). Aflatoxin M1 in raw, pasteurized and powdered milk available in the Syrian market. Food Control, 20, 603–605.

Ghazani, M. H. M. (2009). Aflatoxin M1 contamination in pasteurized milk in Tabriz (northwest of Iran). Food and Chemical Toxicology, 47, 1624–1625.

Gizachew, D., Szonyi, B., Tegegne, A., Hanson, J., & Grace, D. (2016). Aflatoxin contamination

TE D

1105

SC

1103

1110

of milk and dairy feeds in the Greater Addis Ababa milk shed, Ethiopia. Food Control, 59,

1111

773–779.

1114

EP

1113

Golge, O. (2014). A survey on the occurrence of aflatoxin M1 in raw milk produced in Adana province of Turkey. Food Control, 45, 150–155. Gonçalez, G., Felicio, J. D., Pinto, M. M., Rossi, M. H., Nogueira, J. H. C., & Manginelli, S.

AC C

1112

1115

(2005). Ocorrência de aflatoxina M1 em leite bovino comercializado em alguns municípios

1116

do estado de São Paulo. Arquivos do Instituto Biológico, 72, 435–438.

1117

Govaris, A., Roussi, V., Koidis, P. A., & Botsoglou, N. A. (2001). Distribution and stability of

1118

aflatoxin M1 during processing, ripening and storage of Telemes cheese. Food Additives and

1119

Contaminants, 18, 437–443.

ACCEPTED MANUSCRIPT 48 1120

Govaris, A, Roussi, V., Koidis, P. A, & Botsoglou, N. A. (2002). Distribution and stability of

1121

aflatoxin M1 during production and storage of yoghurt. Food Additives and Contaminants,

1122

19, 1043–1050.

1128 1129 1130 1131 1132 1133 1134 1135 1136 1137

RI PT

SC

1127

Cheeses in Turkey. International Journal of Food Properties, 17, 273–282.

Gul, O., & Dervisoglu, M. (2014). Occurrence of aflatoxin M1 in vacuum packed kashar cheeses in Turkey. International Journal of Food Properties, 17, 273-282.

M AN U

1126

Gul, O., & Dervisoglu, M. (2013). Occurrence of Aflatoxin M1 in Vacuum Packed Kashar

Guo, Y., Yuan, Y., & Yue, T. (2013). Aflatoxin M1 in milk products in china and dietary risk assessment. Journal of Food Protection, 76, 849–853.

Gürbay, A., Aydin, S., Girgin, G., Engin, A. B., & Şahin, G. (2006). Assessment of aflatoxin M1 levels in milk in Ankara, Turkey. Food Control, 17, 1–4.

TE D

1125

Technology, 57, 99–109.

Gürses, M., Erdoǧan, A., & Çetin, B. (2004). Occurrence of aflatoxin M1 in some cheese types sold in Erzurum, Turkey. Turkish Journal of Veterinary and Animal Sciences, 28, 527–530. Han, R. W., Zheng, N., Wang, J. Q., Zhen, Y. P., Xu, X. M., & Li, S. L. (2013). Survey of

EP

1124

Guinee, T. P. (2004). Salting and the role of salt in cheese. International Journal of Dairy

aflatoxin in dairy cow feed and raw milk in China. Food Control, 34, 35–39. Hassanin, N. I. (1994). Stability of aflatoxin M1 during manufacture and storage of yoghurt,

AC C

1123

1138

yoghurt-cheese and acidified milk. Journal of the Science of Food and Agriculture, 65, 31–

1139

34.

1140

Haskard, C. A., El-Nezami, H. S., Kankaanpãã, P. E., Salminen, S., & Ahokas, J. T. (2001).

1141

Surface binding of aflatoxin B1 by lactic acid bacteria. Applied and Environmental

1142

Microbiology, 67, 3086-3091.

ACCEPTED MANUSCRIPT 49 1143

Henry, S. H., Whitaker, T., Rabbani, I., Bowers, J., Park, D., Price, W., Bosch, F. X., Pennington, J., Verger, P., Yoshizawa, T., van Egmond, H., Jonker, M. A., & Coker, R. Aflatoxin M1.

1145

IPCS INCHEM, JECFA 47. (2001).

1146

http://www.inchem.org/documents/jecfa/jecmono/v47je02.htm. Accessed 13.10.15.

1147

RI PT

1144

Hernández-Martínez, R., & Navarro-Blasco, I. (2015). Surveillance of aflatoxin content in dairy cow feedstuff from Navarra (Spain). Animal Feed Science and Technology, 200, 35–46.

1149

Hernandez-Mendoza, A., Guzman-de-Peña, D., & Garcia, H. S. (2009). Key role of teichoic acids

SC

1148

on aflatoxin B1 binding by probiotic bacteria. Journal of Applied Microbiology, 107, 395-

1151

403.

1153 1154 1155 1156

Heshmati, A., & Milani, J. M. (2010). Contamination of UHT milk by aflatoxin M1 in Iran. Food Control, 21, 19–22.

Hussain, I., & Anwar, J. (2008). A study on contamination of aflatoxin M1 in raw milk in the Punjab province of Pakistan. Food Control, 19, 393–395.

TE D

1152

M AN U

1150

Hussain, I., Anwar, J., Munawar, M. A., & Asi, M. R. (2008). Variation of levels of aflatoxin M1 in raw milk from different localities in the central areas of Punjab, Pakistan. Food Control,

1158

19, 1126–1129.

1160 1161 1162 1163 1164

Hussain, I., Anwar, J., Asi, M. R., Munawar, M. A., & Kashif, M. (2010). Aflatoxin M1 contamination in milk from five dairy species in Pakistan. Food Control, 21, 122–124.

AC C

1159

EP

1157

Hussein, H. S., & Brasel, J. M. (2001). Toxicity, metabolism and impact of mycotoxins on human and animals. Toxicology, 167, 101–134. Huwig, A., Freimund, S., Käppeli, O., & Dutler, H. (2001). Mycotoxin detoxication of animal feed by different adsorbents. Toxicology Letters, 122, 179-188.

ACCEPTED MANUSCRIPT 50 1165

IARC – International Agency for Research on Cancer. IARC Monograph on the Evaluation of

1166

Carcinogenic Risk to Humans. 82, 171. (2002).

1167

http://monographs.iarc.fr/ENG/Monographs/vol82/mono82.pdf. Accessed 25.10.15.

1169

Iha, M. H., Barbosa, C. B., Okada, I. A., & Trucksess, M. W. (2011). Occurrence of aflatoxin M1 in dairy products in Brazil. Food Control, 22, 1971–1974.

RI PT

1168

Iha, M. H., Barbosa, C. B., Okada, I. A., & Trucksess, M. W. (2013). Aflatoxin M1 in milk and

1171

distribution and stability of aflatoxin M1 during production and storage of yoghurt and

1172

cheese. Food Control, 29, 1–6.

INCHEM – International Program on Chemical Safety – Chemical Safety Information from

M AN U

1173

SC

1170

1174

Intergovernmental Organizations. (1979).

1175

http://www.inchem.org/documents/ehc/ehc/ehc011.htm. Accessed 13.10.15.

1176

Iqbal, S. Z., Asi, M. R., & Ariño, A. (2011). Aflatoxin M1 contamination in cow and buffalo milk samples from the North West Frontier Province (NWFP) and Punjab provinces of Pakistan.

1178

Food Additives and Contaminants – Part B, 4, 282–288.

1180 1181

Iqbal, S. Z., & Asi, M. R. (2013). Assessment of aflatoxin M1 in milk and milk products from Punjab, Pakistan. Food Control, 30, 235–239.

EP

1179

TE D

1177

Iqbal, S. Z., Asi, M. R., & Jinap, S. (2013). Variation of aflatoxin M1 contamination in milk and milk products collected during winter and summer seasons. Food Control, 34, 714–718.

1183

ISIRI - Institute of Standards and Industrial Research of Iran. (2005). Milk and milk products-raw

AC C

1182

1184

milk-specifications and test methods (2nd revision). Iranian National Standard 164. ISIRI:

1185

Karaj, Iran.

1186 1187

Jalili, M., & Scotter, M. (2015). A review of aflatoxin M1 in liquid milk. Iranian Journal of Health, Safety and Environment, 2, 283-295.

ACCEPTED MANUSCRIPT 51 1188 1189

Jasutiene, I., Garmiene, G., & Kulikauskiene, M. (2006). Pasteurisation and fermentation effects on Aflatoxin M1 stability. Milchwissenschaft, 61, 75-79. Jay, J. M. (2005). Microbiologia de Alimentos. Porto Alegre: Artmed.

1191

Joannis-Cassan, C., Tozlovanu, M., Hadjeba-Medjdoub, K., Ballet, N., & Pfohl-Leszkowicz, A.

1192

(2011). Binding of zearalenone, aflatoxin B1, and ochratoxin A by yeast-based products: a

1193

method for quantification of adsorption performance. Journal of Food Protection, 74, 1175–

1194

1185.

SC

1195

RI PT

1190

Josephs, R. D., Ulberth, F., Van Egmond, H.P., & Emons, H. (2005). Aflatoxin M1 in milk powders: Processing, homogeneity and stability testing of certified reference materials.

1197

Food Additives and Contaminants., 22, 864-874.

1198

M AN U

1196

Kabak, B., & Var, I. (2008). Factors affecting the removal of aflatoxin M1 from food model by Lactobacillus and Bifidobacterium strains. Journal of Environmental Science and Health –

1200

Part B, 43, 617–624.

1201

TE D

1199

Kabak, B., & Ozbey, F. (2012). Aflatoxin M1 in UHT milk consumed in Turkey and first assessment of its bioaccessibility using an in vitro digestion model. Food Control, 28, 338–

1203

344.

1205 1206 1207 1208 1209

Kamkar, A. (2005). A study on the occurrence of aflatoxin M1 in raw milk produced in Sarab city of Iran. Food Control, 16, 593–599.

AC C

1204

EP

1202

Kamkar, A. (2006). A study on the occurrence of aflatoxin M1 in Iranian Feta cheese. Food Control, 17, 768–775.

Kamkar, A., Karim, G., Aliabadi, F. S., & Khaksar, R. (2008). Fate of aflatoxin M1 in Iranian white cheese processing. Food and Chemical Toxicology, 46, 2236–2238.

ACCEPTED MANUSCRIPT 52 1210

Kamkar, A., Khaniki, G. R. J., & Alavi, S. A. (2011). Occurrence of Aflatoxin M1 in Raw Milk

1211

Produced in Ardebil of Iran. Iranian Journal of Environmental Health, Science and

1212

Engineering, 8, 123–128. Kaniou-Grigoriadou, I., Eleftheriadou, A., Mouratidou, T., & Katikou, P. (2005). Determination

RI PT

1213 1214

of aflatoxin M1 in ewe’s milk samples and the produced curd and Feta cheese. Food

1215

Control, 16, 257–261.

1218

SC

1217

Kav, K., Col, R., & Kaan Tekinsen, K. (2011). Detection of aflatoxin M1 levels by ELISA in white-brined Urfa cheese consumed in Turkey. Food Control, 22, 1883–1886. Kocasari, F. S., Tasci, F., & Mor, F. (2012). Survey of aflatoxin M1 in milk and dairy products

M AN U

1216

1219

consumed in Burdur, Turkey. International Journal of Dairy Technology, 65, 365–371.

1220

Kos, J., Lević, J., Duragić, O., Kokić, B., & Miladinović, I. (2014). Occurrence and estimation of

1221

aflatoxin M1 exposure in milk in Serbia. Food Control, 38, 41–46. Lee, J. E., Kwak, B.-M., Ahn, J.-H., & Jeon, T.-H. (2009). Occurrence of aflatoxin M1 in raw

1223

milk in South Korea using an immunoaffinity column and liquid chromatography. Food

1224

Control, 20, 136–138.

TE D

1222

Lindner, E. (1995). Toxicologia de los Alimentos. Zaragoza: Acribia AS.

1226

López, C., Ramos, L., Ramadán, S., Bulacio, L., & Perez, J. (2001). Distribution of aflatoxin M1

1227

in cheese obtained from milk artificially contaminated. International Journal of Food

1228

Microbiology, 64, 211–215.

1230

AC C

1229

EP

1225

López, C. E., Ramos, L. L., Ramadán, S. S., & Bulacio, L. C. (2003). Presence of aflatoxin M1 in milk for human consumption in Argentina. Food Control, 14, 31–34.

1231

Malissiova, E., Tsakalof, A., Arvanitoyannis, I. S., Katsafliaka, A., Katsioulis, A., Tserkezou, P.,

1232

Koureas, M., Govaris, A., & Hadjichristodoulou, C. (2013). Monitoring Aflatoxin M1 levels

ACCEPTED MANUSCRIPT 53 1233

in ewe’s and goat's milk in Thessaly, Greece; potential risk factors under organic and

1234

conventional production schemes. Food Control, 34, 241–248.

1235

Manetta, A. C., Giammarco, M., Giuseppe, L. Di, Fusaro, I., Gramenzi, A., Formigoni, A., Vignola, G., & Lambertini, L. (2009). Distribution of aflatoxin M1 during Grana Padano

1237

cheese production from naturally contaminated milk. Food Chemistry, 113, 595–599.

1238

Markaki, P., & Melissari, E. (1997). Occurrence of aflatoxin M1 in commercial pasteurized milk

1240

determined with ELISA and HPLC. Food Additives and Contaminants, 14, 451–6.

SC

1239

RI PT

1236

Martins, M. L., & Martins, H. M. (2000). Aflatoxin M1 in raw and ultra high temperature-treated milk commercialized in Portugal. Food Additives and Contaminants, 17, 871–874.

1242

Martins, M. L., & Martins, H. M. (2004). Aflatoxin M1 in yoghurts in Portugal. International

1243 1244

M AN U

1241

Journal of Food Microbiology, 91, 315–317.

Martins, H. M., Guerra, M. M., & Bernardo, F. (2005). A six year survey (1999-2004) of the ocurrence of aflatoxin M1 in daily products produced in Portugal. Micotoxin Research, 21,

1246

192-195.

1248 1249

Masoero, F., Gallo, A., Moschini, M., Piva, G., & Diaz, D. (2007). Carryover of aflatoxin from feed to milk in dairy cows with low or high somatic cell counts. Animal, 1, 1344–1350.

EP

1247

TE D

1245

Masoero, F., Gallo, A., Diaz, D., Piva, G., & Moschini, M. (2009). Effects of the procedure of inclusion of a sequestering agent in the total mixed ration on proportional aflatoxin M1

1251

excretion into milk of lactating dairy cows. Animal Feed Science and Technology, 150, 34-

1252

45.

1253 1254 1255

AC C

1250

Mendonça, C., & Venâncio, A. (2005). Fate of aflatoxin M1 in cheese whey processing. Journal of the Science of Food and Agriculture, 85, 2067–2070. Midio, A. F., & Martins, D. I. (2000). Toxicologia de Alimentos. São Paulo: Varela.

ACCEPTED MANUSCRIPT 54 1256 1257 1258

Ministry of Health of the People’s Republic of China. (2011). National food safety standard: maximum levels of mycotoxins in foods. GB2761-2011. Beijing: Ministry of Health. Mohajeri, F. A., Ghalebi, S. R., Rezaeian, M., Gheisari, H. R., Azad, H. K., Zolfaghari, A., & Fallah, A. A. (2013). Aflatoxin M1 contamination in White and Lighvan cheese marketed in

1260

Rafsanjan, Iran. Food Control, 33, 525-527.

RI PT

1259

Mohamadi Sani, A., Khezri, M., & Moradnia, H. (2012). Determination of Aflatoxin M1 in Milk

1262

by ELISA Technique in Mashad (Northeast of Iran). International Scholarly Research

1263

Notices: Toxicology, 2012, 1–4.

Mohammadi, H., Alizadeh, M., Bari, M. R., Khosrowshahi, A., & Tadjik, H. (2009).

M AN U

1264

SC

1261

1265

Optimization of the process variables for minimizing of the aflatoxin M1 content in Iranian

1266

white brine cheese. Journal of Agricultural Science and Technology, 11, 181–190.

1267

Mohammadian, B., Khezri, M., Ghasemipour, N., Mafakheri, Sh., & Poorghafour Langroudi, P. (2010). Aflatoxin M1 contamination of raw and pasteurized milk produced in Sanandaj, Iran.

1269

Archives of Razi Institute, 65, 99–104.

1270

TE D

1268

Montagna, M. T., Napoli, C., De Giglio, O., Iatta, R., & Barbuti, G. (2008). Occurrence of aflatoxin M1 in dairy products in Southern Italy. International Journal of Molecular

1272

Sciences, 9, 2614–2621.

1274

Motawee, M. M., & McMahon, D. J. (2009). Fate of aflatoxin M1 during manufacture and

AC C

1273

EP

1271

storage of feta cheese. Journal of Food Science, 74, 42–45.

1275

Motawee, M. M. (2013). Reduction of Aflatoxin M1 Content during Manufacture and Storage of

1276

Egyptian Domaiti Cheese. International Journal of Veterinary Medicine: Research and

1277

Reports, 2013, 1-10.

ACCEPTED MANUSCRIPT 55 1278

Moosavy, M. H., Roostaee, N., Katiraee, F., Habibi-Asl, B., Mostafavi, H., & Dehghan, P.

1279

(2013). Aflatoxin M1 occurrence in pasteurized milk from various dairy factories in Iran.

1280

International Food Research Journal, 20, 3351–3355.

1283 1284 1285

update. Journal of Food Science, 71, 51–65.

RI PT

1282

Murphy, P. A., Hendrich, S., Landgren, C., & Bryant, C. M. (2006). Food mycotoxins: An

Mulunda, M., & Mike, D. (2014). Occurrence of aflatoxin M1 from rural subsistence and commercial farms from selected areas of South Africa. Food Control, 39, 92–96.

SC

1281

Nachtmann, C., Gallina, S., Rastelli, M., Ferro, G. L., & Decastelli, L. (2007). Regional monitoring plan regarding the presence of aflatoxin M1 in pasteurized and UHT milk in

1287

Italy. Food Control, 18, 623–629.

1288

M AN U

1286

Nakajima, M., Tabata, S., Akiyama, H., Itoh, Y., Tanaka, T., Sunagawa, H., Tyonan, T., Yoshizawa, T. & Kumagai, S. (2004). Occurrence of aflatoxin M1 in domestic milk in Japan

1290

during the winter season. Food Additives and Contaminants, 21, 472–478.

1293 1294 1295 1296 1297 1298 1299

aflatoxin M1 in milk samples in Ardabil, Iran. Food Control, 21, 1022–1024. Nilchian, Z., & Rahimi, E. (2012). Aflatoxin Ml in yoghurts, cheese and ice-cream in

EP

1292

Nemati, M., Mehran, M. A., Hamed, P. K., & Masoud, A. (2010). A survey on the occurrence of

Shahrekord-Iran. World Applied Sciences Journal, 19, 621–624. Oatley, J. T., Rarick, M. D., Ji, G. E., & Linz, J. E. (2000). Binding of aflatoxin B1 to

AC C

1291

TE D

1289

bifidobacteria in vitro. Journal of Food Protection, 63, 1133–1136. Oliveira, C. A. F., & Ferraz, J. C. O. (2007). Occurrence of aflatoxin M1 in pasteurised, UHT milk and milk powder from goat origin. Food Control, 18, 375–378. Oliveira, C. A. F., Franco, R. C., Rosim, R. E., & Fernandes, A. M. (2011). Survey of aflatoxin

1300

M1 in cheese from the North-east region of São Paulo, Brazil. Food Additives and

1301

Contaminants – Part B, 4, 57–60.

ACCEPTED MANUSCRIPT 56 1302

Oliveira, C. P. de, Soares, N. D. F. F., Oliveira, T. V. de Baffa Júnior, J. C., & Silva, W. A.

1303

(2013a). Aflatoxin M1 occurrence in ultra high temperature (UHT) treated fluid milk from

1304

Minas Gerais/Brazil. Food Control, 30, 90–92. Oliveira, C. A. F.; Bovo, F.; Corassin, C. H.; Jager, A. V.; & Reddy, K. R. (2013b). Recent trends

1306

in microbiological decontamination of aflatoxins in foodstuffs. In M. Razzaghi-Abyaneh

1307

(Ed.), Aflatoxins - Recent Advances and Future Prospects (pp. 59-92). Rijeka: InTech.

1308

Oruc, H. H., Cibik, R., Yilmaz, E., & Kalkanli, O. (2006). Distribution and stability of Aflatoxin

SC

RI PT

1305

M1 during processing and ripening of traditional white pickled cheese. Food Additives and

1310

Contaminants, 23, 190–195.

1312 1313 1314 1315

Oruc, H. H., Cibik, R., Yilmaz, E., & Gunes, E. (2007). Fate of aflatoxin M1 in kashar cheese. Distribution, 27, 82–90.

Oveisi, M. R., Jannat, B., Sadeghi, N., Hajimahmoodi, M., & Nikzad, A. (2007). Presence of aflatoxin M1 in milk and infant milk products in Tehran, Iran. Food Control, 18, 1216–1218.

TE D

1311

M AN U

1309

Payne, G. A. (1998). Process of contamination by aflatoxin-producing fungi and their impact on crops. In K. K. Sinha, & D. Bhatnagar (Eds.), Mycotoxins in agriculture and food safety.

1317

New York: Marcel Dekker.

1319 1320

Peitri, A., Bertuzzi, T., Bertuzzi, P., & Piva, G. (2009). Aflatoxin M1 occurrence in samples of Grana Padano cheese. Food Additives and Contaminants, 14, 341–4.

AC C

1318

EP

1316

Peltonem, K., El-Nezami, H., Haskard, C., Ahokas, J., & Salminen, S. (2001). Aflatoxin B1

1321

binding by dairy strains of lactic acid bacteria and bifidobacteria. Journal of Dairy Science,

1322

84, 2152–2156.

1323

Pereira, M. M. G., Carvalho, E. P., Prado, G., Rosa, C. A. R., Veloso, T., Souza, L. A. F., &

1324

Ribeiro, J. M. M. (2005). Aflatoxinas em alimentos destinados a bovinos e em amostras de

1325

leite da região de Lavras, Minas Gerais - Brasil. Ciência e Agrotecnologia, 29, 106-112.

ACCEPTED MANUSCRIPT 57 Picinin, L. C. A., Cerqueira, M. M. O. P., Vargas, E. A., Lana, A. M. Q., Toaldo, I. M., &

1327

Bordignon-Luiz, M. T. (2013). Influence of climate conditions on aflatoxin M1

1328

contamination in raw milk from Minas Gerais State, Brazil. Food Control, 31, 419–424.

1329

Pierides, M., El-Nezami, H., Peltonen, K., Salminen, S., & Ahokas, J. (2000). Ability of dairy

1330

strains of lactic acid bacteria to bind aflatoxin M1 in a food model. Journal of Food

1331

Protection, 63, 645–650.

1334

review. Nutrition Research, 15, 767-776.

SC

1333

Piva, G., Galvano, F., Pietri, A., & Piva, A. (1995). Detoxification methods of aflatoxins. A

Pleadin, J., Vulić, A., Perši, N., Škrivanko, M., Capek, B., & Cvetnić, Ž. (2015). Annual and

M AN U

1332

RI PT

1326

1335

regional variations of aflatoxin B1 levels seen in grains and feed coming from Croatian dairy

1336

farms over a 5-year period. Food Control, 47, 221–225.

1337

Prado, G., Oliveira, M. S., Abrantes, F. M., Santos, L. G., Soares, C. R., & Veloso, T. (1999). Ocorrência de aflatoxina M1 em leite consumido na cidade de Belo Horizonte - Minas

1339

Gerais / Brasil - agosto/98 à abril/99. Ciência e Tecnologia de Alimentos, 19, 420–423.

1340

Prado, G., Oliveira, M. S., Carvalho, E. P., Veloso, T., Sousa, L. A. F., & Cardoso, A. C. F.

TE D

1338

(2001). Aflatoxina M1 em queijo prato e parmesão determinada por coluna de

1342

imunoafinidade e cromatografia líquida. Revista do Instituto Adolfo Lutz, 60, 147–151. Prandini, A., Tansini, G., Sigolo, S., Filippi, L., Laporta, M., & Piva, G. (2009). On the

AC C

1343

EP

1341

1344

occurrence of aflatoxin M1 in milk and dairy products. Food and Chemical Toxicology, 47,

1345

984–991.

1346 1347 1348 1349

Purchase, I. F. H., Steyn, M., Rinsma, R., & Tustin, R. C. (1972). Reduction of the aflatoxin M content of milk by processing. Food and Cosmetics Toxicology, 10, 383–387. Rahaie, S., Emam-Djomeh, Z., Razavi, S. H., & Mazaheri, M. (2012). Evaluation of aflatoxin decontaminating by two strains of Saccharomyces cerevisiae and Lactobacillus rhamnosus

ACCEPTED MANUSCRIPT 58 1350

strain GG in pistachio nuts. International Journal of Food Science and Technology, 47,

1351

1647-1653.

1352

Rastogi, S., Dwivedi, P. D., Khanna, S. K., & Das, M. (2004). Detection of Aflatoxin M1 contamination in milk and infant milk products from Indian markets by ELISA. Food

1354

Control, 15, 287–290.

1356 1357

Riahi-Zanjani, B., & Balali-Mood, M. (2013). Aflatoxin M1 contamination in commercial

pasteurized milk from local markets in Fariman, Iran. Mycotoxin Research, 29, 271–274.

SC

1355

RI PT

1353

Rodríguez Velasco, M. L., Calonge Delso, M. M., & Ordónez Escudero, D. (2003). ELISA and HPLC determination of the occurrence of aflatoxin M(1) in raw cow’s milk. Food Additives

1359

and Contaminants, 20, 276–280.

1360

M AN U

1358

Rokhi, M. L., Darsanaki, R. K., Mohammadi, M., Kolavani, M. H., Issazadeh, K., Aliabadi, M. A., & Branch, L. (2013). Determination of Aflatoxin M1 Levels in Raw Milk Samples in

1362

Gilan , Iran. Advances Studies in Biology, 5, 151–156.

1363

TE D

1361

Roussi, V., Govaris, A., Varagouli, A., & Botsoglou, N. A. (2002). Occurrence of aflatoxin M(1) in raw and market milk commercialized in Greece. Food Additives and Contaminants, 19,

1365

863–868.

EP

1364

Ruangwises, N., & Ruangwises, S. (2010). Aflatoxin M1 contamination in raw milk within the

1367

central region of Thailand. Bulletin of Environmental Contamination and Toxicology, 85,

1368

195–198.

1369

AC C

1366

Rubio, R., Moya, V. J., Berruga, M. I., Molina, M. P., & Molina, A. (2011). Aflatoxin M1 in the

1370

intermediate dairy products from Manchego cheese production: distribution and stability.

1371

Mljekarstvo, 61, 283–290.

1372 1373

Rustom, I. Y. S. (1997). Aflatoxin in food and feed: occurence, legislation and inactivation by physical methods. Food Chemistry, 59, 57–67.

ACCEPTED MANUSCRIPT 59 1374 1375 1376

Sadia, A., Jabbar, M. A., Deng, Y., Hussain, E. A., Riffat, S., Naveed, S., & Arif, M. (2012). A survey of aflatoxin M1 in milk and sweets of Punjab, Pakistan. Food Control, 26, 235–240. Şanli, T., Deveci, O., & Sezgin, E. (2012). Effects of Pasteurization and Storage on Stability of Aflatoxin M1 in Yogurt. Kafkas Üniversitesi Veteriner Fakültesi Dergisi, 18, 987–990.

1378

Santini, A., Raiola, A., Ferrantelli, V., Giangrosso, G., Macaluso, A., Bognanno, M., Galvano, F.,

1379

& Ritieni, A. (2013). Aflatoxin M1 in raw, UHT milk and dairy products in Sicily (Italy).

1380

Food Additives and Contaminants – Part B, 6, 181–186.

1383 1384 1385 1386 1387

SC

Abyaneh (Ed.), Aflatoxins – Recent advances and future prospects. InTech.

M AN U

1382

Santini, A., & Ritieni, A. (2013). Aflatoxins: risk, exposure and remediation. In M. Razzaghi-

Sarimehmetoglu, B., Kuplulu, O., & Haluk Celik, T. (2004). Detection of aflatoxin M1 in cheese samples by ELISA. Food Control, 15, 45–49.

Sarimehmetoğlu, B., & Küplülü, Ö. (2004). Binding ability of aflatoxin M1 to yoghurt bacteria. Ankara Üniversitesi Veteriner Fakültesi Dergisi,, 51, 195–198.

TE D

1381

RI PT

1377

Sassahara, M., Pontes Netto, D., & Yanaka, E. K. (2005). Aflatoxin occurrence in foodstuff supplied to dairy cattle and aflatoxin M1 in raw milk in the North of Paraná state. Food and

1389

Chemical Toxicology, 43, 981–984.

1391 1392

Sengun, I. Y., Yaman, D. B., & Gonul, S. A. (2008). Mycotoxins and mould contamination in cheese: a review. World Mycotoxin Journal, 1, 291–298.

AC C

1390

EP

1388

Shetty, P. H., & Jespersen, L. (2006). Saccharomyces cerevisiae and lactic acid bacteria as

1393

potential mycotoxin decontaminating agents. Trends Food Science and Technology, 17, 48–

1394

55.

1395

Shetty, P. H., Hald, B., & Jespersen, L. (2007). Surface binding of aflatoxin B1 by

1396

Saccharomyces cerevisiae strains with potential decontaminating abilities in indigenous

1397

fermented foods. International Journal of Food Microbiology, 113, 41–46.

ACCEPTED MANUSCRIPT 60 1398

Shundo, L., Navas, S. A., Lamardo, L. C. A., Ruvieri, V., & Sabino, M. (2009). Estimate of

1399

aflatoxin M1 exposure in milk and occurrence in Brazil. Food Control, 20, 655–657.

1400

Siddappa, V., Nanjegowda, D. K., & Viswanath, P. (2012). Occurrence of aflatoxin M1 in some samples of UHT, raw & pasteurized milk from Indian states of Karnataka and Tamilnadu.

1402

Food and Chemical Toxicology, 50, 4158–4162.

1403

RI PT

1401

Silva, R. A., Chalfoun, S. M., Silva, M. A. M., & Pereira, M. C. (2007). Inquérito sobre o

consumo de alimentos possíveis de contaminação por micotoxinas na ingesta alimentar de

1405

escolares da cidade de Lavras, MG. Ciência e Agrotecnologia,, 31, 439-447. Simas, M. M. S., Botura, M. B., Correa, B., Sabino, M., Mallmann, C. A., Bitencourt, T. C. B. S.

M AN U

1406

SC

1404

1407

C., & Batatinha, M. J. M. (2007). Determination of fungal microbiota and mycotoxins in

1408

brewers grain used in dairy cattle feeding in the State of Bahia, Brazil. Food Control, 18,

1409

404–408.

Škrbić, B., Živančev, J., Antić, I., & Godula, M. (2014). Levels of aflatoxin M1 in different types

TE D

1410 1411

of milk collected in Serbia: Assessment of human and animal exposure. Food Control, 40,

1412

113–119.

Srivastava, V. P., Bu-Abbas, A., Alaa-Basuny, Al-Johar, W., Al-Mufti, S., & Siddiqui, M. K. J.

EP

1413

(2001). Aflatoxin M1 contamination in commercial samples of milk and dairy products in

1415

Kuwait. Food Additives and Contaminants, 18, 993–997.

1416

AC C

1414

Souza, S. V. C., Vargas, E. A., & Junqueira, R. G. (1999). Eficiência de um kit de ELISA na

1417

detecção e quantificação de aflatoxina M1 em leite e investigação da ocorrência no estado de

1418

Minas Gerais. Ciência e Tecnologia de Alimentos, 19, 401-405.

1419

Suriyasathaporn, W., & Nakprasert, W. (2012). Seasonal patterns of aflatoxin M1 contamination

1420

in commercial pasteurised milk from different areas in Thailand. Food Additives and

1421

Contaminants - Part B, 5, 145–149.

ACCEPTED MANUSCRIPT 61 1422 1423 1424

Tamine, A. Y., & Robinson, R. K. (2007). Tamine and Robinson’s Yoghurt: Science and technology. Cambridge: Woodhead Publishing and CRC Press LLC. Tajik, H., Rohani, S. M. R., & Moradi, M. (2007). Detection of aflatoxin M1 in raw and commercial pasteurized milk in Urmia, Iran. Pakistan Journal of Biological Sciences, 10,

1426

4103–4107.

1427

RI PT

1425

Tajkarimi, M., Shojaee Aliabadi, F., Salah Nejad, M., Pursoltani, H., Motallebi, A. A., &

Mahdavi, H. (2007). Seasonal study of aflatoxin M1 contamination in milk in five regions in

1429

Iran. International Journal of Food Microbiology, 116, 346–349.

Tajkarimi, M., Aliabadi-Sh, F., Salah Nejad, A., Poursoltani, H., Motallebi, A. A., & Mahdavi,

M AN U

1430

SC

1428

1431

H. (2008). Aflatoxin M1 contamination in winter and summer milk in 14 states in Iran. Food

1432

Control, 19, 1033–1036.

Tavakoli, H. R., Riazipour, M., Kamkar, A., Shaldehi, H. R., & Mozaffari Nejad, A. S. (2012).

1434

Occurrence of aflatoxin M1 in white cheese samples from Tehran, Iran. Food Control, 23,

1435

293–295.

1438 1439 1440 1441 1442

consumed in Turkey. Food and Chemical Toxicology, 46, 3287–3289.

EP

1437

Tekinşen, K. K., & Eken, H. S. (2008). Aflatoxin M1 levels in UHT milk and kashar cheese

Türk Gida Kodeksi Teblig. (2002). Resmi Gazete, 23 Eylül 2002, Sayı: 24885. Ankara: Başbakanlık Basmevi.

AC C

1436

TE D

1433

Unusan, N. (2006). Occurrence of aflatoxin M1 in UHT milk in Turkey. Food and Chemical Toxicology, 44, 1897–1900. US FDA - Food and Drug Administration. (1996). Sec.527. 400 whole milk, low fat milk, skim

1443

milk-aflatoxin M1 (CPG 7106.210). In FDA compliance policy guides (pp.219).

1444

Washington.

ACCEPTED MANUSCRIPT 62 1445

Van Egmond, H. P., Paulsch, W. E., Veringa, H. A., & Schuller, P. L. (1977). The effect of

1446

processing on the Aflatoxin M1 content of milk and milk products. Archives de l’Institut

1447

Pasteur de Tunis, 54, 381–390.

1449 1450

Van Egmond, H. P., & Jonker, M. A. (2004). Worldwide regulations on aflatoxins – the situation in 2002. Journal of Toxicology - Toxin Reviews, 23, 273-293.

RI PT

1448

Van Egmond, H. P., Schothorst, R. C., & Jonker, M. A. (2007). Regulations relating to

mycotoxins in food - Perspectives in a global and European context. Analytucal and

1452

Bioanalytical Chemistry, 389, 147–157.

1454 1455 1456

Var, I., & Kabak, B. (2009). Detection of aflatoxin M1 in milk and dairy products consumed in

M AN U

1453

SC

1451

Adana, Turkey. International Journal of Dairy Technology, 62, 15–18. Varga, J., Frisvad, J. C., & Samson, R. A. (2009). A reappraisal of fungi producing aflatoxins. World Mycotoxin Journal, 2, 263-277.

Virdis, S., Corgiolu, G., Scarano, C., Pilo, A. L., & De Santis, E. P. L. (2008). Occurrence of

1458

Aflatoxin M1 in tank bulk goat milk and ripened goat cheese. Food Control, 19, 44–49.

1459

Walstra, P., Wouters, J. T. M., & Geurts, T. J. (2006). Dairy Science and Technology. Boca

1462 1463 1464 1465 1466

EP

1461

Raton: Taylor & Francis.

Wiseman, D. W., & Marth, E. H. (1983). Behavior of Aflatoxin M1 in Yoghurt, Buttermilk and Kefir. Journal of Food Protection, 46, 115–118.

AC C

1460

TE D

1457

Wu, Q., Jezkova, A., Yuan, Z., Pavlikova, L., Dohnal, V., & Kuca, K. (2009). Biological degradation of aflatoxins. Drug Metabolism Reviews, 41, 1-7. Yaroglu, T., Oruc, H. H., & Tayar, M. (2005). Aflatoxin M1 levels in cheese samples from some provinces of Turkey. Food Control, 16, 883–885.

ACCEPTED MANUSCRIPT 63 1467

Yousef, A. E., & Marth, E. H. (1989). Stability and degradation of Aflatoxin M1. In H. P. van

1468

Egmond (Ed.), Mycotoxin in dairy products (pp. 127-161). London and New York: Elsevier

1469

Applied Science. Zheng, N., Sun, P., Wang, J. Q., Zhen, Y. P., Han, R. W., & Xu, X. M. (2013). Occurrence of

1471

aflatoxin M1 in UHT milk and pasteurized milk in China market. Food Control, 29, 198–

1472

201.

RI PT

1470

Zinedine, A., González-Osnaya, L., Soriano, J. M., Moltó, J. C., Idrissi, L., & Mañes, J. (2007).

1474

Presence of aflatoxin M1 in pasteurized milk from Morocco. International Journal of Food

1475

Microbiology, 114, 25–29.

1477

Zinedine, A., & Mañes, J. (2009). Occurrence and legislation of mycotoxins in food and feed from Morocco. Food Control, 20, 334–344.

1478

1483 1484 1485 1486 1487 1488 1489 1490

EP

1482

AC C

1481

TE D

1479 1480

M AN U

1476

SC

1473

ACCEPTED MANUSCRIPT 64 1491

Figure Captions

1492

1494 1495 Figure 2: Structures of the main aflatoxins. 1496

RI PT

1493 Figure 1: Overview of biotransformation pathways for aflatoxin B1 (with permission).

SC

1497 Figure 3: Basic steps involved in the production of pasteurized (A) and UHT (B) milk. 1498

M AN U

1499 Figure 4: Basic steps involved in the production of cheese (A) and yogurt (B). 1500 1501 1502

EP AC C

1504

TE D

1503

ACCEPTED MANUSCRIPT Table 1: Maximum limits for AFM1 in milk and dairy products in different countries. Country

Milk (µg/kg)

Dairy Products (µg/kg)

Reference

USA

0.05

0.50

US FDA (1996)

EU

0.050

0.050

European Commission (2001);

0.050

Iran

0.50 (milk powder)

RI PT

European Commission, (2006) ISIRI (2005)

0.020 (butter and butter milk)

SC

0.250 (cheese) 0.050

Turkey

0.250 (cheese)

Codex Alimentarius Commission

0.50

Brazil

M AN U

(2001); Türk Gda Kodeksi Teblig

5 (milk powder)

(2002) Anvisa (2011)

2.5 (cheese)

0.050

0.5

EP

China

0.05

AC C

Pakistan

0.250 (soft cheese)

Anfossi et al. (2011); Galvano et

0.450 (hard cheese)

al. (2001)

TE D

Italy

0.5 (milk products)

Ministry of Health of the People’s Republic of China (2011)

0.050

Iqbal et al. (2011) Iqbal and Asi (2013)

Switzerland

0.050

0.250 (cheese)

Creppy (2002); Fallah (2010a)

The Netherland

0.050

0.020 (butter)

Creppy (2002); Fallah (2010a)

* EU Regulation 466/2001.

0.020 (cheese)

ACCEPTED MANUSCRIPT 1 1

Table 2: Incidence of AFM1 in milk and dairy products, analytical methods and limit of detection

2

(LOD)/limit of quantification (LOQ). Type of

Technique/

Number of

Number

product

Instrument

samples

of positive

LOD/LOQ1

Reference

RI PT

Country

samples Antigen–

56

6

Powdered milk

antibody

5

4

Pasteurized

reaction

16

milk Brazil

UHT Milk

LC

Pasteurized Milk

Milk

Brazil

Raw Milk Pasteurized

Brazil

Brazil

HPLC

AC C

Milk

HPLC

Raw Milk

TLC

1

/n.s.

López et al. (2003)

8

60

53

15 ng L-1

Garrido et al.

79

58

/50 ng L-1

(2003)

43

17

0.5 ng mL-

Gonçalez et al.

TE D

Pasteurized

EP

Brazil

0.01 µg L-

SC

Milk

M AN U

Argentina

1

/n.s.

(2005)

36

19

2 ng L-1/6

Pereira et al.

34

13

ng L-1

(2005)

42

10

245 ng L-

Sassahara et al.

1

/n.s.

(2005)

40

40

3 ng kg-1/10

Shundo et al.

Pasteurized

10

7

ng kg-1

(2009)

Milk

75

72

UHT Milk

HPLC-FLD

ACCEPTED MANUSCRIPT 2 Powdered Milk Brazil

UHT Milk

Immunoaffinity

75

150 ng L-

23

1

Brazil

Raw Milk

ELISA

129

129

HPLC

/n.s.

(2013a)

0.0003 µg

Picinin et al.

L-1/n.s.

(2013)

RI PT

columns

Oliveira et al.

0.0001 µg

SC

L-1/0.0002 µg L-1

Raw Cow Milk

ELISA

China

UHT Milk

ELISA

Pasteurized

Croatia

ELISA

Fresh Milk

ELISA

EP

Croatia

Milk

Bulk Cow Milk

ELISA

Han et al. (2013)

153

84

0.005 µg L-

Zheng et al.

26

25

233

61

(2013)

5 ng kg-

Guo et al. (2013)

Bilandžić et al.

/2.8 ng L-1

(2010)

23.2 ng L-

Bilandžić et al.

1

32

2

Raw Sheep

19

0

Milk

14

0

/n.s.

1.1 ng L-

61

Raw Goat Milk

Milk

/n.s.

1

59

Raw Donkey

1

112

337

AC C

China

5 ng L-1/n.s.

TE D

Milk

65

200

M AN U

China

1

/35.8 ng L1

(2014a)

ACCEPTED MANUSCRIPT 3 Greece

Commercial

ELISA

Pasteurized

HPLC

81

5 ng L-1/n.s.

72

Markaki and Melissari (1997)

Milk 30

22

Raw Goat Milk

10

4

Raw Sheep

12

8

Milk

82

70

Pasteurized

17

Milk

15

UHT Milk Concentrated Milk Bulk Cow Milk

ELISA

234

TE D

Greece

food

ELISA

EP

Infant Milk

AC C

Infant formula

Roussi et al. (2002)

14 14

43

HPLC

India

5 ng L-1/n.s.

RI PT

LC

SC

Raw Cow Milk

M AN U

Greece

0.004 µg

Malissiova et al.

kg-1/0.0125

(2013)

µg kg-1

17

17

18

17

40

38

12

4

21

21

n.s./n.s.

Rastogi et al. (2004)

Milk based

Cereal weaning food

Liquid Milk India

UHT (plain

HPLC

2.1 µg L-

Siddappa et al.

ACCEPTED MANUSCRIPT 4 Milk)

24

9

111

85

1

/n.s.

(2012)

0.015 µg L-

Kamkar (2005)

UHT (flavored

Iran

Raw Milk

TLC

RI PT

Milk)

1

Pasteurized

EIA/Ridascreen

Milk

aflatoxin M1

624

Test Pasteurized

ELISA

Milk Infant Formula Iran

Raw Milk

ELISA

Iran

Iran

Raw Milk

Raw Milk

HPLC

AC C

Iran

EP

Milk

Pasteurized

HPLC

128

120

116

72

72

72

72

TE D

Pasteurized

128

M AN U

Iran

624

98

n.s./n.s.

SC

Iran

/n.s.

98

5 ng kg1

Alborzi et al. (2006)

Oveisi et al.

/n.s.

(2007)

n.s./n.s.

Tajik et al. (2007)

0.0003 µg

Tajkarimi et al.

L-1/0.001

(2007)

µg L-1 319

172

n.s./0.005

Tajkarimi et al.

µg kg-1

(2008)

ELISA

50

50

5 ng L-1/n.s.

Ghazani (2009)

ELISA

210

116

0.005 µg L-

Heshmati and

Milk

Iran

UHT Milk

1

/n.s.

Milani (2010)

ACCEPTED MANUSCRIPT 5

Iran

Pasteurized

ELISA/Ridascr

116

83

Milk

een aflatoxin

109

68

UHT Milk

M1 Test

Pasteurized

TLC

91

66

ELISA

240

226

32

31

Pasteurized

ELISA

Milk

ELISA

Raw Cow Milk

Raw Sheep

Iran

AC C

Milk

TLC

EP

Raw Goat Milk

Pasteurized

90

122

TE D

Iran

Raw Milk

M AN U

Milk

Iran

0.012 µg L-

Fallah (2010b)

ELISA

/n.s.

n.s./n.s.

SC

Raw Milk Pasteurized

Iran

Fallah (2010a)

1

Milk Iran

5 ng L-1/n.s.

RI PT

Iran

90

122

5 ng kg1

Pasteurized Milk

ELISA

al. (2010)

Nemati et al.

/n.s.

(2010)

n.s./n.s.

Kamkar et al. (2011)

88

74

0.0125 µg

Fallah et al.

65

28

L-1/n.s.

(2011)

72

43

42

41

n.s./n.s.

Mohamadi Sani et

milk

Iran

Mohammadian et

al. (2012) 80

77

n.s./n.s.

Moosavy et al. (2013)

ACCEPTED MANUSCRIPT 6 Iran

Raw Milk

ELISA

90

56

n.s./n.s.

Rokhi et al. (2013)

Pasteurized

ELISA

45

UHT milk

HPLC

Raw Milk

ELISA

161

296 (2004)

Italy

Fresh

HPLC

pasteurized and UHT Milk Raw Milk

HPLC

Bulk Milk UHT Milk

HPLC

EP

Kuwait

UHT Milk

Milk and Milk

AC C

Japan

316

HPLC

Fresh Cow Milk

ELISA

2

n.s./n.s.

Galvano et al. (2001)

Decastelli et al. (2007) Nachtmann et al. (2007)

3 ng L-1/4

Santini et al.

10

3

ng L-1

(2013)

12

5

208

207

0.001 µg

Nakajima et al.

kg-1/n.s.

(2004)

0.01 µg L-

Srivastava et al.

54

14

1

177

176

105

-

27

-

UHT Cow Milk

5 ng L-1/n.s.

0

Products

Kuwait

1 ng L-1/n.s.

(2013)

27

51

TE D

Italy

5

M AN U

45 (2005)

125

SC

Italy

Riahi-Zanjani and Balali-Mood

Milk

Italy

5 ng L-1/n.s.

45

RI PT

Iran

/n.s.

(2001)

5 ng kg-

Dashti et al.

1

/n.s.

(2009)

ACCEPTED MANUSCRIPT 7 Powdered Milk (Baby formula) Pasteurized

HPLC

54

48 1

Milk

Pakistan

Raw Cow Milk

Raw Cow Milk

HPLC

AflaTest

48

168

column/fluoro meter Pakistan

Raw Buffalo

HPLC

Milk

Pakistan

Buffalo Milk

Goat Milk

AC C

Sheep Milk

HPLC

EP

Cow Milk

Camel Milk

Pakistan

Buffalo Milk

HPLC

Cow Milk

Pakistan

Milk

360

153

120

63

TE D

Raw Cow Milk

ELISA

168

M AN U

affinity

13

Zinedine et al.

/3.9 ng L-1

(2007)

n.s./n.s.

El Marnissi et al.

n.s./n.s.

SC

Morocco

1.0 ng L-

RI PT

Morocco

0.004 µg L1

(2012)

Hussain and

Anwar (2008)

Hussain et al.

/n.s.

(2008)

n.s./n.s.

Hussain et al.

55

19

40

15

30

6

24

4

20

0

94

46

0.004 µg

84

42

kg-1/n.s.

232

177

0.002 µg L-

(2010)

1

/n.s.

Iqbal et al. (2011)

Sadia et al. (2012)

ACCEPTED MANUSCRIPT 8 Pakistan

Raw Cow Milk

HPLC

107

76

n.s./n.s.

Iqbal and Asi (2013)

HPLC

UHT Milk Portugal

Raw Milk

HPLC

UHT Milk Portugal

Raw Milk

HPLC

104

39

84

35

31

25

70

60

598

394

0.004 µg L1

ELISA

Milk UHT Milk Raw,

UHPLC/HESI-

Pasteurized and

MS/MS

Sterilized Milk Serbia

Cow Milk

ELISA

EP

Goat Milk Donkey Milk

Africa South Africa

Bulk Milk

AC C

South

0

36

17

50

38

TE D

Serbia

4

M AN U

Pasteurized

HPLC

/n.s.

0.005 µg L1

Martins and

/n.s.

Martins (2000)

0.005 µg L-

Martins et al.

1

Portugal

Iqbal et al. (2013)

RI PT

Raw Cow Milk

SC

Pakistan

/n.s.

(2005)

5 ng L-1/n.s.

Duarte et al. (2013)

0.0002 µg

Škrbić et al.

kg-1/0.0007

(2014)

µg kg-1 1.50 ng kg-

150

148

10

8

5

3

kg-1

90

90

n.s./n.s.

1

Kos et al. (2014)

/5.00 ng

Dutton et al. (2012)

Raw Milk

Fluorometry

from rural

125

107/125

Mulunda and

TLC

26/125

Mike (2014)

subsistence

HPLC

98/125

(RSFs)

Fluorometry

100

82/100

0.01 ng mL1

/n.s.

ACCEPTED MANUSCRIPT 9 Raw Milk

TLC

15/100

from

HPLC

85/100 0.01 ng mL-

commercial

(CDFs) Raw Cow Milk

HPLC

100

ELISA

9

72

TE D

Raw Milk

VICAM

EP

Syria

ELISA

35

3

1

Velasco et al.

n.s./10 ng L-

(2003)

68

35

5 ng kg-

11

7

Raw Goat Milk

10

10

8

1

Pasteurized

Cano-Sancho et

/n.s.

al. (2010)

0.02 µg kg-

Ali et al. (2014)

1

Milk

Powdered Milk

1

1

13

Cow Milk

1

Rodríguez

23

Raw Sheep

/0.02 µg L-

n.s./10 ng L-

70

ELISA

Lee et al. (2009)

5

74

Raw Milk

AC C

Sudan

UHT Milk

92

M AN U

Raw Cow Milk

HPLC

Spain

0.002 µg L-

1

Korea

Spain

48

SC

South

/n.s.

RI PT

1

dairy farms

/n.s.

n.s./n.s.

Ghanem and Orfi (2009)

ACCEPTED MANUSCRIPT 10 Turkey

Raw Milk

TLC

90

79

0.0125 µg

Bakirci (2001)

L-1/n.s. Pasteurized

ELISA

85

75

ELISA/Ridascr

129

75

Milk

UHT Milk

1

een aflatoxin M1 Test

Turkey

UHT Milk

HPLC

Raw Milk

ELISA/Ridascr een aflatoxin

Raw Milk

Raw Cow Milk

40

1

67

Unusan (2006)

/n.s.

10 ng L-

24

8

Çelik et al. (2005)

Gürbay et al.

/n.s.

(2006)

10 ng L-

Tekinşen and

1

/n.s.

Eken (2008)

0.004 µg L-

Kabak and Ozbey

1

/0.014 µg

(2012)

L-1

45

41

45

30

176

53

5 ng L-1/n.s.

Kocasari et al. (2012)

M1 Test

AC C

Milk

Thailand

100

UHT Milk

Pasteurized

Turkey

ELISA

27

M AN U

Turkey

HPLC

TE D

Turkey

UHT Milk

EP

Turkey

0.01 µg L-

SC

Turkey

n.s./n.s.

RI PT

Turkey

HPLC

0.021 µg

Golge (2014)

kg-1/0.025 µg kg-1 HPLC

240

240

n.s./0.01 µg

Ruangwises and

L-1

Ruangwises

ACCEPTED MANUSCRIPT 11 (2010) Thailand

Pasteurized

HPLC

120

0.004 µg L-

-

1

Minas Frescal

HPLC

cheese

24

6

24

7

Minas Padrão Cheese Minas Frescal

LC

cheese Yoghurt Dairy Drink

Iran

Feta Cheese

TLC

Yoghurt

TLC

1

(2012)

1

0.01 ng g-

Oliveira et al.

/0.03 ng g-1

(2011)

47

12

10

80

66

3 ng kg-1/10

Iha et al. (2011)

ng kg-1

0.015 µg L1

Kamkar (2006)

/n.s.

0.012 µg L-

68

45

72

59

36

25

31

8

75

49

0.0125 µg

Fallah et al.

Cheese

61

30

L-1/n.s.

(2011)

Industrial

60

14

Yoghurt

64

34

Traditional

61

19

Ice cream

AC C

Butter

EP

White Cheese

Iran

49

53

TE D

Iran

58

M AN U

Brazil

and Nakprasert

SC

Brazil

/0.01 µg L-

RI PT

Milk

Suriyasathaporn

Lighvan

TLC

1

Fallah (2010b)

/n.s.

ACCEPTED MANUSCRIPT 12 Yoghurt

71

16

Industrial

65

9

Kashk

RI PT

Traditional Kashk Industrial

SC

Doogh

Doogh Iran

Yoghurt

ELISA

Cheese Ice cream

Iran

White Cheese

White Cheese

Cheese

Grana Padano

AC C

Italy

ELISA

EP

Lighvan

ELISA

40

14

40

16

40

12

50

30

TE D

Iran

M AN U

Traditional

HPLC

Yoghurt

Rahimi (2012)

n.s./n.s.

45

29

37

10

223

219

n.s./n.s.

Dry milk for

Tavakoli et al.

120

73

92

49

94

12

Mohajeri et al. (2013)

5 ng kg1

HPLC

Nilchian and

(2012)

cheese

Italy

n.s./n.s.

Peitri et al. (1997)

/n.s.

1 ng L-1/n.s.

Galvano et al. (2001)

infant formula Italy

Sheep cheese

HPLC

0.037 µg

Montagna et al.

ACCEPTED MANUSCRIPT 13 92

25

Buffalo cheese

51

0

Sheep-goat

16

5

cheese

12

2

102

85

Goat cheese Italy

Cheese

ELISA

kg-1/n.s.

25 ng kg1

HPLC

Milk cream

Pakistan

Cheese

Cheese

ELISA

Yoghurt

3 ng L-1/4

Santini et al.

3

ng L-1

(2013)

40

32

50 ng kg-

Dashti et al.

111

75

70

35

79

27

119

93

Cheese Cream

150

89

Yoghurt

96

59

Butter

74

33

AC C

White Cheese

HPLC-

1

/n.s.

(2009)

50 ng L-

Elkak et al.

1

32

Ice cream

HPLC

7

96

Butter

Pakistan

ELISA

TE D

Lebanon

7

EP

Kuwait

17

(2012)

SC

Cheese

Anfossi et al.

/n.s.

M AN U

Italy

(2008)

RI PT

Cow cheese

/n.s.

(2012)

0.004 µg L-

Iqbal et al.

1

/n.s.

(2013)

n.s./n.s.

Iqbal and Asi (2013)

ACCEPTED MANUSCRIPT 14 Portugal

Natural

HPLC

48

10 ng kg-

2

1

Yoghurt with

48

16

72

0

72

2

pieces of

Cheese

ELISA

Yoghurt Turkey

White Cheese

ELISA

Kashar Cheese

100

Tulum Cheese Processed Cheese

85

100

79

23

9

EIA

Kashar cheese

(Ridascreen®

14

6

Tulum cheese

Aflatoxin)

11

7

9

4

6

2

200

10

Kashar Cheese

200

12

Processed

200

8

94

86

49

44

EP

Lor cheese

ELISA

AC C

White Cheese

25 ng kg1

Martins (2004)

Cano-Sancho et

/n.s.

al. (2010)

n.s./n.s.

Sarimehmetoğlu

81

White cheese

Civil cheese

Turkey

82

100

TE D

Turkey

100

M AN U

Spain

SC

strawberries

/n.s.

RI PT

Yoghurt

Martins and

n.s./n.s.

et al. (2004)

Gürses et al. (2004)

100 ng kg1

Yaroglu et al.

/n.s.

(2005)

625 ng kg-

Aycicek et al.

Cheese

Turkey

White Cheese Cheese

ELISA

1

/n.s.

(2005)

ACCEPTED MANUSCRIPT 15

Turkey

Kashar Cheese

53

47

Butter

27

25

132

109

Kashar Cheese

ELISA

50 ng L1

Turkish white

ELISA

70

5

ELISA

127

36

ELISA

White Cheese

ELISA

Kashar Cheese

Yoghurt

14

20

8

20

16

50

28

50

26

AC C

Turkey

White Cheese

ELISA

/n.s.

5 ng kg-

1

Kav et al. (2011)

/n.s.

50 ng kg-

Ardic et al.

/n.s.

(2009)

50 ng kg-

Ertas et al. (2011)

1

/n.s.

5 ng kg1

EP

Dairy dessert

159

20

TE D

Tulum Cheese

193

M AN U

Turkish white brinedcheese

Turkey

Er et al. (2010)

1

Urfa cheese Turkey

0.05 µg kg-

SC

White-brined

Eken (2008)

1

brined cheese Turkey

/n.s.

RI PT

Turkey

Tekinsen and

/n.s.

5 ng kg1

45

42

Butter

45

39

Yoghurt

45

20

Ice cream

45

34

Milk powder

45

42

/n.s.

50 ng kg1

/n.s.

25 ng kg1

/n.s.

50 ng kg-

Kocasari et al. (2012)

ACCEPTED MANUSCRIPT 16 1

/n.s.

10 ng kg1

/n.s.

RI PT

50 ng kg1

Turkey

Kashar cheese

HPLC

147

144

0.01 µg kg-

3 1n.s.: not specified.

M AN U

4 5 6 7

13 14 15 16 17 18

EP

12

AC C

11

TE D

8

10

/n.s.

SC

1

9

/n.s.

Gul and

Dervisoglu (2014)

AC C

EP

TE D

M AN U

SC

RI PT

ACCEPTED MANUSCRIPT

AC C

EP

TE D

M AN U

SC

RI PT

ACCEPTED MANUSCRIPT

AC C

EP

TE D

M AN U

SC

RI PT

ACCEPTED MANUSCRIPT

AC C

EP

TE D

M AN U

SC

RI PT

ACCEPTED MANUSCRIPT