The Presence of Aflatoxin M1 in Milk and Milk Products in Bangladesh

As milk provides both micro- and macronutrients, it is an important component in the diet. However, the presence of aflatoxin B₁ (AFB₁) in the feed of dairy cattle results in contamination of milk and dairy products with aflatoxin M₁ (AFM₁), a toxic metabolite of the carcinogenic mycotoxin. With the aim to determine AFM₁ concentrations in milk and milk products consumed in Bangladesh, in total, 145 samples were collected in four divisional regions (Sylhet, Dhaka, Chittagong, and Rajshahi). The samples comprised these categories: raw milk (n = 105), pasteurized milk (n = 15), ultra-high temperature (UHT)-treated milk (n = 15), fermented milk products such as yogurt (n = 5), and milk powder (n = 5). AFM₁ levels in these samples were determined through competitive enzyme-linked immunosorbent assay (ELISA). Overall, AFM₁ was present in 78.6% of milk and milk products in the range of 5.0 to 198.7 ng/L. AFM₁ was detected in 71.4% of raw milk (mean 41.1, range 5.0–198.7 ng/L), and in all pasteurized milk (mean 106, range 17.2–187.7 ng/L) and UHT milk (mean 73, range 12.2–146.9 ng/L) samples. Lower AFM₁ levels were found in yogurt (mean 16.9, range 8.3–41.1 ng/L) and milk powder samples (mean 6.6, range 5.9–7.0 ng/L). About one-third of the raw, pasteurized, and UHT milk samples exceeded the EU regulatory limit (50 ng/L) for AFM₁ in milk, while AFM₁ levels in yogurt and milk powder samples were well below this limit. Regarding regions, lower AFM₁ contamination was observed in Chittagong (mean 6.6, max 10.6 ng/L), compared to Sylhet (mean 53.7, max 198.7 ng/L), Dhaka (mean 37.8, max 97.2 ng/L), and Rajshahi (mean 34.8, max 131.4 ng/L). Yet, no significant difference was observed in AFM₁ levels between summer and winter season. In conclusion, the observed frequency and levels of aflatoxin contamination raise concern and must encourage further monitoring of AFM₁ in milk and milk products in Bangladesh.     

Aspergillus flavus and Total Aflatoxins Occurrence in Dairy Feed and Aflatoxin M1 in Bovine Milk in Aguascalientes,Mexico

Contamination of food chains by toxigenic fungi and aflatoxins is a global problem that causes damage to human health, as well as to crop and livestock production. The objective is to evaluate Aspergillus flavus and total aflatoxins (AFs) occurrence in totally mixed rations (TMRs) for dairy cows and aflatoxin M₁ (AFM₁) in milk for human consumption. Ninety-nine dairy production units located in Aguascalientes, Mexico, were randomly selected, and samples were collected from TMRs, raw milk, and milk marketed in the city in two consecutive agricultural cycles. AFs were quantified in TMRs and milk by indirect enzyme immunoassay and HPLC; aflatoxigenic and molecular (PCR) capacity of monosporic A. flavus isolates in the feed was characterized. All feed, raw, and pasteurized milk samples showed aflatoxin contamination (26.0 ± 0.4 µg/kg, 32.0 ± 1.0, and 31.3 ± 0.7 ng/L, respectively), and a significant proportion (90.4, 11.3, and 10.3%) exceeded the locally applied maximum permissible limits for feed and milk (20.0 µg/kg and 50 ng/L). Aflatoxin contamination in both TMRs and milk indicated a seasonal influence, with a higher concentration in the autumn–winter cycle when conditions of higher humidity prevail. The results obtained suggest the existence of contamination by aflatoxigenic A. flavus and aflatoxins in the diet formulated for feeding dairy cows and, consequently, in the dairy food chain of this region of the Mexican Highland Plateau.     

Adsorbents Reduce Aflatoxin M1 Residue in Milk of Healthy Dairy Cow Exposed to Moderate Level Aflatoxin B1 in Diet and Its Exposure Risk for Humans

This study investigated the effect of moderate risk level (8 µg/kg) AFB₁ in diet supplemented with or without adsorbents on lactation performance, serum parameters, milk AFM₁ content of healthy lactating cows and the AFM₁ residue exposure risk in different human age groups. Forty late healthy lactating Holstein cows (270 ± 22 d in milk; daily milk yield 21 ± 3.1 kg/d) were randomly assigned to four treatments: control diet without AFB₁ and adsorbents (CON), CON with 8 μg/kg AFB₁ (dry matter basis, AF), AF + 15 g/d adsorbent 1 (AD1), AF + 15 g/d adsorbent 2 (AD2). The experiment lasted for 19 days, including an AFB₁-challenge phase (day 1 to 14) and an AFB₁-withdraw phase (day 15 to 19). Results showed that both AFB₁ and adsorbents treatments had no significant effects on the DMI, milk yield, 3.5% FCM yield, milk components and serum parameters. Compared with the AF, AD1 and AD2 had significantly lower milk AFM₁ concentrations (93 ng/L vs. 46 ng/L vs. 51 ng/L) and transfer rates of dietary AFB₁ into milk AFM₁ (1.16% vs. 0.57% vs. 0.63%) (p < 0.05). Children aged 2–4 years old had the highest exposure risk to AFM₁ in milk in AF, with an EDI of 1.02 ng/kg bw/day and a HI of 5.11 (HI > 1 indicates a potential risk for liver cancer). Both AD1 and AD2 had obviously reductions in EDI and HI for all population groups, whereas, the EDI (≥0.25 ng/kg bw/day) and HI (≥1.23) of children aged 2–11 years old were still higher than the suggested tolerable daily intake (TDI) of 0.20 ng/kg bw/day and 1.00 (HI). In conclusion, moderate risk level AFB₁ in the diet of healthy lactating cows could cause a public health hazard and adding adsorbents in the dairy diet is an effective measure to remit AFM₁ residue in milk and its exposure risk for humans.     

AFM1 in Milk: Physical,Biological, and Prophylactic Methods to Mitigate Contamination

Aflatoxins (AFs) are toxic, carcinogenic, immunosuppressive secondary metabolites produced by some Aspergillus species which colonize crops, including many dietary staple foods and feed components. AFB₁ is the prevalent and most toxic among AFs. In the liver, it is biotransformed into AFM₁, which is then excreted into the milk of lactating mammals, including dairy animals. AFM₁ has been shown to be cause of both acute and chronic toxicoses. The presence of AFM₁ in milk and dairy products represents a worldwide concern since even small amounts of this metabolite may be of importance as long-term exposure is concerned. Contamination of milk may be mitigated either directly, decreasing the AFM₁ content in contaminated milk, or indirectly, decreasing AFB₁ contamination in the feed of dairy animals. Current strategies for AFM₁ mitigation include good agricultural practices in pre-harvest and post-harvest management of feed crops (including storage) and physical or chemical decontamination of feed and milk. However, no single strategy offers a complete solution to the issue.     

Assessment of aflatoxin M1 levels in pasteurised milk,raw milk,and cheese in Arak,Iran

Aflatoxins are fungal toxins, which may be found in food and due to negative health effects, they are of major concerns for human health and food industries. Aflatoxin M₁ contamination in dairy products in Arak, Iran, 2013 was evaluated using ELISA method. Sample groups were comprised of 111 samples, including pasteurized milk (n?=?56), raw milk (n?=?16) and cheese (n?=?39), and were analyzed for AfM₁ content. The results showed that 93.7% of the samples were positive with the total average concentration of 85.8?µg/L in milk and 30.39?µg/kg in cheese. Moreover, AfM₁ of 94.4% of milk samples and 92.3% of cheese samples was above the standards of Iran and EU.     

Occurrence of Staphylococcus aureus on Farms with Small Scale Production of Raw Milk Cheeses in Poland

This paper describes the results of a 3-year study on the prevalence, enterotoxinogenicity and resistance to antimicrobials of S. aureus isolated on dairy farms with small scale production of raw cow milk cheeses. The samples of raw milk, semi-finished products and the final products as well as swabs were collected between 2011 and 2013 from nine dairy farms in Poland. A total of 244 samples were examined, of which 122 (50.0%) were contaminated with S. aureus including 18 of 26 (69.2%) mature cheese samples with log10 CFU g⁻¹ between <1- and 7.41. In swabs collected from the staff and production environment the highest contamination rate with coagulase positive staphylococci (CPS) was detected on hands of cheese makers (4.34 log10 CFU/swab). None of the cheese samples contaminated with CPS contained staphylococcal enterotoxins (SEs). However, 55 of 122 (45.1%) S. aureus isolates possessed SEs genes, mainly (26 of 55; 47.3%) a combination of the sed, sej and ser genes. Furthermore, the sep (15 of 55; 27.3%) as well as seg and sei (9 of 55; 16.4%) genes were also identified. The remaining S. aureus isolates possessed the sea gene (one isolate), the combination of sec, seg and sei (three isolates) as well as the sed, sej, sep and ser markers together (one CPS). Resistance to penicillin (62 of 122 isolates; 50.8%) was the most common among the tested isolates. Some CPS were also resistant to chloramphenicol (7; 5.7%) and tetracycline (5; 4.1%). The obtained results indicated that the analyzed cheeses were safe for consumers. To improve the microbiological quality of traditional cheese products more attention should be paid to animal welfare and hygiene practices during the process of cheese manufacturing in some dairy farms.     

Determination of Urinary Biomarkers for Assessment of Short-Term Human Exposure to Aflatoxins in S?o Paulo,Brazil

In the present study, a longitudinal assessment was carried out to evaluate the short-term human exposure to aflatoxins in Pirassununga region, São Paulo, Brazil, by determination of urinary aflatoxins by a liquid chromatography coupled to mass sprectrometry (UPLC-MS/MS) method. Sixteen volunteers with ages ranging from 14 to 55 years old were instructed to collect the early morning first urine four times every three months, from June 2011 to March 2012, totaling 64 samples. Aflatoxin M₁ (AFM₁) was found in 39 samples (61%) at levels ranging from 0.19 to 12.7 pg·mg⁻¹ creatinine (mean: 1.2 ± 2.0 pg·mg⁻¹ creatinine). Residues of aflatoxins B₁, B₂, G₁, G₂ and aflatoxicol were not identified in any urine sample. No significant difference was found among the AFM₁ mean levels in urine samples collected in the four sampling periods. The levels of AFM₁ found in urine samples indicate a low short-term exposure of the population studied to aflatoxins through the diet, although further investigations are needed to assess other long-term biomarkers of exposure to AFB₁.     

Occurrence of aflatoxin M1 in some samples of UHT,raw & pasteurized milk from Indian states of Karnataka and Tamilnadu

Aflatoxin M₁ (AFM₁) is a toxic metabolite found in the milk of lactating animals which have consumed feedstuffs contaminated with aflatoxin B₁. Ultra high temperature treated (UHT) milk is a product which is becoming popular in developing countries like India as there is a lack of proper cold storage or refrigeration facilities. In this study, 45 samples of UHT milk of popular brands prevalent in the market were analyzed for the presence of AFM₁ by reversed phase HPLC using fluorescent detector after cleanup of sample with immunoaffinity columns. All samples of plain UHT milk were positive for AFM₁ and 38% of these contained levels more than 0.5 μg/kg, the maximum permitted limit prescribed by the Codex Alimentarius Commission and by the mandatory regulations of the country, the FSSAI Regulations, 2011. In 62.5% of flavored UHT milk, AFM₁ was below detectable levels (0.02 μg L⁻¹). However, 12.5% of these samples also contained levels exceeding the maximum permitted limits. AFM₁ was present in 61.6% of the 52 raw milk samples analyzed from the two states of Karnataka and Tamilnadu with a range of 0.1–3.8 μg L⁻¹. 17.3% of these samples also exceeded the regulatory limits of the country.     

Analysis of Mycotoxin and Secondary Metabolites in Commercial and Traditional Slovak Cheese Samples

Cheese represents a dairy product extremely inclined to fungal growth and mycotoxin production. The growth of fungi belonging to Aspergillus, Penicillium, Fusarium, Claviceps, Alternaria, and Trichoderma genera in or on cheese leads to undesirable changes able to affect the quality of the final products. In the present investigation, a total of 68 types of commercial and traditional Slovak cheeses were analyzed to investigate the occurrence of fungal metabolites. Altogether, 13 fungal metabolites were identified and quantified. Aflatoxin M1, the only mycotoxin regulated in milk and dairy products, was not detected in any case. However, the presence of metabolites that have never been reported in cheeses, such as tryptophol at a maximum concentration level from 13.4 to 7930 µg/kg (average: 490 µg/kg), was recorded. Out of all detected metabolites, enniatin B represents the most frequently detected mycotoxin (0.06–0.71 µg/kg) in the analyzed samples. Attention is drawn to the lack of data on mycotoxins’ origin from Slovak cheeses; in fact, this is the first reported investigation. Our results indicate the presence of fungal mycotoxin contamination for which maximum permissible levels are not established, highlighting the importance of monitoring the source and producers of contamination in order to protect consumers’ health.     

Aflatoxin M1 in Cow’s Milk: Method Validation for Milk Sampled in Northern Italy

Aflatoxins (AFs) are mycotoxins produced by some species of Aspergillus. In dairy cows, ingested AFB1 is metabolized into carcinogenic AFM1 which is eliminated through milk, thus posing a risk for consumer health. Here we describe the set, validation, and application of screening (ELISA) and confirmatory (HPLC) tests carried out on milk samples collected through official control of mycotoxin levels in northern Italy over a three-year period (2012–2014). The limit of detection (LOD) was set at 5 ppt (ng/kg) and 2 ppt for ELISA and HPLC, respectively, and the limit of quantification (LOQ) was 10 ppt for confirmatory HPLC. A total of 1668 milk samples were analyzed: ELISA identified 36 (2.2%) positive milk samples that were subsequently confirmed by HPLC. The level of AFM1 in the positive samples ranged between 18 ± 2 and 208 ± 27 ppt. Of the total samples, only eight (0.5%) were found non-compliant with the EU regulatory limit (50 ppt; range 74 ± 10 to 208 ± 27 ppt). Use of ELISA and HPLC tests in series allows for high-volume analysis of samples, thus saving time and money while guaranteeing high analytical precision and accuracy.     

Effectiveness of Training and Use of Novasil Binder in Mitigating Aflatoxins in Cow Milk Produced in Smallholder Farms in Urban and Periurban Areas of Kenya

Aflatoxins, which commonly contaminate animal feeds and human food, present a major public health challenge in sub-Saharan Africa. After ingestion by cows, aflatoxin B1 is metabolized to aflatoxin M1 (AFM1), some of which is excreted in milk. This study involved smallholder dairy farms in urban and periurban areas of Nairobi and Kisumu, Kenya. The objective was to determine the effectiveness of training and providing farmers with aflatoxin binder (NovaSil^®) on AFM1 contamination in raw milk. A baseline survey was undertaken and 30 farmers whose milk had AFM1 levels above 20 ppt were randomly selected for inclusion in the study. Of these, 20 farmers were part of the intervention, and were given training on the usage of the NovaSil^® binder, while 10 served as a control group. All farmers were visited biweekly for three months for interviews and milk samples were collected to measure the AFM1 levels. The AFM1 levels were quantified by enzyme linked immunosorbent assay. The NovaSil^® binder significantly reduced AFM1 concentrations in the raw milk produced by the farmers in the intervention group over the duration of the study (p < 0.01). The control farms were more likely to have milk with AFM1 levels exceeding the regulatory limit of 50 ppt compared to the intervention farms (p < 0.001) (odds ratio = 6.5). The farmers in the intervention group perceived that there was an improvement in milk yield, and in cow health and appetite. These farmers also felt that the milk they sold, as well as the one they used at home, was safer. In conclusion, the use of binders by dairy farmers can be effective in reducing AFM1 in milk. Further research is needed to understand their effectiveness, especially when used in smallholder settings.     

Composition of Asarum heterotropoides var. mandshuricum radix oil from different extraction methods and activities against human body odor-producing bacteria

In this study, oils from Asarum heterotropoides were extracted by traditional solvent extraction and supercritical CO₂ (SC-CO₂) extraction methods and their antioxidant activities along with antimicrobial and inhibitory activities against five human body odor-producing bacteria (Staphylococcus epidermidis, Propionibacterium freudenreichii, Micrococcus luteus, Corynebacterium jeikeium, and Corynebacterium xerosis) were evaluated. The oil was found to contain 15 components, among which the most abundant component was methyl eugenol (37.6%), which was identified at every condition studied in different extraction methods. The oil extracted with n-hexane and ethanol mixture exhibited a strong antioxidant activity (92% ± 2%) and the highest ABTS and 2,2-diphenyl-1-picrylhydrazyl scavenging activities (89% ± 0.2%). The highest amounts of total phenolic content and total flavonoid content were 23.1 ± 0.4 mg/g and 4.9 ± 0.1 mg/g, respectively, in the traditional method. In the SC-CO₂ method performed at 200 bar/50°C using ethanol as an entrainer, the highest inhibition zone was recorded against all the aforementioned bacteria. In particular, strong antibacterial activity (38 ± 2 mm) was found against M. luteus. The minimum inhibitory concentration (MIC) for the oil against bacteria ranged from 10.1 ± 0.1 μg/mL to 46 ± 2 μg/mL. The lowest MIC was found against M. luteus. Methyl eugenol was found to be one of the major compounds working against human body odor-producing bacteria.     

Sodium Butyrate More Effectively Mitigates the Negative Effects of High-Concentrate Diet in Dairy Cows than Sodium β-Hydroxybutyrate via Reducing Free Bacterial Cell Wall Components in Rumen Fluid and Plasma

The present study was aimed at investigating the effects of sodium butyrate and sodium β-hydroxybutyrate on lactation and health of dairy cows fed a high-concentrate (HC) diet. Eighty mid-lactation dairy cows with an average milk yield of 33.75 ± 5.22 kg/d were randomly allocated to four groups (n = 20 per group) and were fed either a low-concentrate (LC) diet, a HC diet, the HC diet with 1% sodium butyrate (HCSB), or the HC diet with 1% sodium β-hydroxybutyrate (HCHB). The feeding trial lasted for 7 weeks, with a 2-week adaptation period and a 5-week measurement period, and the trial started from 96 ± 13 d in milk. Sodium butyrate supplementation delayed the decline in milk production and improved milk synthesis efficiency and milk fat content. Additionally, it decreased the proinflammatory cytokines and acute phase proteins (APPs) in plasma, the leucocytes in blood, the somatic cell count (SCC) in milk, and the gene expression of pattern recognition receptors (PRRs) and proinflammatory cytokines in the mammary gland, due to decreasing the contents of bacterial cell wall components (lipopolysaccharide, LPS; peptidoglycan, PGN; and lipoteichoic acid, LTA) in the rumen and plasma, compared with the HC diet. Sodium β-hydroxybutyrate supplementation also improved milk yield, milk synthesis efficiency and milk fat content and partially reduced the adverse effects caused by the HC diet, but it had no effect on decreasing bacterial cell wall components in the rumen and plasma, compared with the HC diet. Collectively, both sodium butyrate and sodium β-hydroxybutyrate mitigated the negative effects of HC diet on lactation and health of dairy cows, with sodium butyrate being more effective than sodium β-hydroxybutyrate.     

Mycotoxin Co-Occurrence in Milks and Exposure Estimation: A Pilot Study in São Paulo,Brazil

The aim of this study was to conduct a first evaluation on the co-occurrence of aflatoxins (AF) M₁, B₁, B₂, G₁ and G₂; fumonisins (F) B₁ and B₂; deoxynivalenol (DON); de-epoxydeoxinivalenol (DOM-1); ochratoxin A (OTA); zearalenone (ZEN); α-zearalenol (α-ZEL); and β-zearalenol (β-ZEL) in 68 samples of fluid milk consumed in Pirassununga, São Paulo, Brazil. The probable daily intake (PDI) was also calculated for each mycotoxin evaluated. Mycotoxins were determined by liquid chromatography coupled to mass spectrometry. Sixty-two (91.2%) samples contained at least one type of mycotoxin. AFM₁ was found in 6 samples (8.8%), and none of them presented concentrations above the Brazilian maximum permitted level in milk (500 ng/L). Low levels of non-regulated mycotoxins DOM-1, OTA, FB₁, FB₂, α-ZEL and β-ZEL were found in 6 (8.8%), 17 (25%), 10 (14.7%), 3 (4.4%), 39 (57.4%) and 28 (41.2%) samples of milk, respectively. None of the PDIs calculated for the quantified mycotoxins were above recommended values, indicating low exposure through milk consumption in the area studied. However, 21 samples (30.9%) contained 2–4 types of mycotoxins, which warrants concern about the potential adverse effects of mycotoxin mixtures in milks.     

Infants exposure to aflatoxin M1 as a novel foodborne zoonosis

Occurrence of aflatoxin M₁ (AFM₁) in infant formula milk powder (IFMP) and maternal breast milk (MBM) was investigated as a risk factor affects the health of newborns in Egypt. A total of 125 IFMP and 125 MBM samples were collected and examined for the presence of AFM₁ using competitive ELISA test. The results indicated that the relative risk (RR) of exposure to AFM₁ via consumption of MBM was higher than IFMP (RR; 1.6, 95% CI; 1.28–2.03, p = 0.0001). The mean concentrations of AFM₁ were significantly differed (p < 0.0001) between MBM (74.413 ± 7.070 ng/l) and IFMP (9.796 ± 1.036 ng/l). High frequency distributions were detected within the range of 5–25 ng/l and >50–100 ng/l in IFMP and MBM, respectively. The average daily exposure of newborns to AFM₁ via consumption of MBM and IFMP was 52.684 and 8.170 ng, respectively, with a significant difference at p < 0.0001. Consumption of raw milk by lactating mothers exhibited a significant correlation (p < 0.0001) with the presence of AFM₁ in their milk. In conclusion, this work established a pioneering concept that AFM₁ may be considered as an etiological factor for a novel foodborne zoonosis identified as Aflatoxicosis M₁.     

On the occurrence of aflatoxin M1 in milk and dairy products

Aflatoxins are toxic fungal metabolites found in foods and feeds. When ruminants eat AFB₁-feedstuffs, they metabolise the toxin and excrete AFM₁ in milk. To control AFM₁ in foods it is necessary to reduce AFB₁ contamination of feeds for dairy cattle by preventing fungal growth and AFB₁ formation in agricultural commodities intended for animal use. Corn and corn-based products are one of the most contaminated feedstuffs; therefore risk factor analysis of AFB₁ contamination in corn is necessary to evaluate risk of AFM₁ contamination in milk and milk products. During the corn silage production, the aflatoxins production is mostly influenced by: harvest time; fertilization; irrigation; pest control; silage moisture; and storage practices. Due to the lower moisture at harvest and to the conservation methods, the corn grain is mostly exposed to the contamination by Aspergillus species. Therefore, it is necessary to reduce the probability of this contaminant through choice of: hybrids; seeding time and density; suitable ploughing and fertirrigation; and chemical or biological control. Grains harvested with the lowest possible moisture and conservation moisture close to or less than 14% are necessary to reduce contamination risks, as is maintaining mass to homogeneous moisture. Kernel mechanical damage, grain cleaning practices and conservation temperature are also factors which need to be carefully controlled.     

Aflatoxin M1 contamination and antibiotic residue in milk in Khorasan province,Iran

The aim of this study was to evaluate aflatoxin M₁ (AFM₁) contamination and antibiotic presence in milk samples in Khorasan province in Iran. During 4 months (March to June 2008), one hundred ninety-six milk samples were collected from seven dairies. The occurrence and concentration range of AFM₁ in the samples were investigated by competitive enzyme-linked immunoabsorbent assay (ELISA) method. Antibiotic presence was determined using Copan test, a broad-spectrum test capable of detecting beta-lactams, tetracyclines, sulfonamides, aminoglycosides and macrolydes. AFM₁ was found in 100% of the examined milk samples by average concentration of 77.92 ng/kg. The concentration of AFM₁ in all of the samples were lower than Iranian national standard and FDA limit (500 ng/L), but 80.6% of the samples had AFM₁ greater than the maximum tolerance limit (50 ng/L) accepted by European Union and Codex Alimentarius Commission. Statistical evaluation showed no significant difference between the mean concentrations of AFM₁ of milk samples taken from different factories (P > 0.05). Copan milk test was positive for 40.8% of the samples.     

Field Monitoring of Aflatoxins in Feed and Milk of High-Yielding Dairy Cows under Two Feeding Systems

Aflatoxin M1 (AFM1) is a hydroxylated metabolite of aflatoxin B1 (AFB1) that can be excreted in milk of cows after consuming contaminated feed. The aim of this study consisted of a field monitoring to assess the contamination levels of AFB1 in 60 feed samples from two feeding systems for high-yielding dairy cows and of AFM1 in the corresponding raw milk samples. The aflatoxins were analyzed by in-house validated methods based on high-performance liquid chromatography (HPLC) with fluorescence detection. AFB1 was detected in 55% of feed samples (mean 0.61 μg/kg, with 2 samples exceeding the European Union (EU) maximum level set at 5 μg/kg), with greater incidence and concentration in compound feed than in unifeed rations (p < 0.05). AFM1 was detected in 38.3% milk samples (mean 12.6 ng/kg, with 5 samples exceeding the EU maximum level set at 50 ng/kg), with a higher occurrence in milk of cows fed compound feed, as well as in spring milk compared to that produced in winter. The overall transfer ratio of aflatoxins from feed to milk was 3.22%, being higher in cows fed with compound feed and in spring milkings. In a selection of positive matched samples (n = 22), the ratio AFM1/AFB1 exceeded the European Food Safety Authority (EFSA) estimated 6% threshold for high-yielding dairy cows.     

Mycobiota and Mycotoxin Contamination of Traditional and Industrial Dry-Fermented Sausage Kulen

The aim of this study was to identify and compare surface mycobiota of traditional and industrial Croatian dry-fermented sausage Kulen, especially toxicogenic species, and to detect contamination with mycotoxins recognized as the most important for meat products. Identification of mould species was performed by sequence analysis of beta- tubulin and calmodulin gene, while the determination of mycotoxins aflatoxin B₁ (AFB₁), ochratoxin A (OTA), and cyclopiazonic acid (CPA) was carried out using the LC-MS/MS (liquid chromatography-tandem mass spectrometry) method. The results showed a significantly higher number of mould isolates and greater species (including of those mycotoxigenic) diversity in traditional Kulen samples in comparison with the industrial ones. P. commune, as a potential CPA-producer, was the most represented in traditional Kulen (19.0%), followed by P. solitum (16.6%), which was the most represented in industrial Kulen samples (23.8%). The results also showed that 69% of the traditional sausage samples were contaminated with either CPA or OTA in concentrations of up to 13.35 µg/kg and 6.95 µg/kg, respectively, while in the industrial samples only OTA was detected (in a single sample in the concentration of 0.42 µg/kg). Mycotoxin AFB₁ and its producers were not detected in any of the analysed samples (<LOD).