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

Abstract

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).

The Occurrence of Five Unregulated Mycotoxins Most Important for Traditional Dry-Cured Meat Products

“…In the recent study of mycotoxin contamination of the traditional sausage Kulen performed by Lešić et al [ 27 ], OTA concentrations in 2 out of 16 samples exceeded the recommended value of 1 µg/kg in pork meat and products adopted by the Italian Ministry of Health (3.91 and 6.95 µg/kg) [ 37 ]. In that research, in most of the OTA-contaminated samples, no OTA-producing mould was identified.…”

“…Although P. commune , a significant CPA producer, is one of the predominant mould species isolated from dry-fermented meat products, CPA concentrations in products of this type are generally unexplored [ 26 , 41 , 42 ]. To the best of our knowledge, data published so far include 1 study of Spanish dry-cured hams, in which CPA concentrations ranged from 36 to 540 µg/kg [ 28 ], and 2 studies of Croatian traditional sausages [ 26 , 27 ], in which CPA was found in 15% and 31% samples, in the concentration range of 2.50–59.80 µg/kg. In comparison with previous research, the results of this study, which included dry-fermented sausages and dry-cured meat products, showed higher CPA concentrations than in the previous two Croatian sausage studies, and slightly lower concentrations than those found in Spanish dry-cured hams.…”

“…Just like OTA, CIT can act as hepatotoxin, teratogen, foetotoxin, and genotoxin, but the IARC still tags it as a Group 3 compound that cannot be classified as a human carcinogen [ 25 ]. A few studies on CPA occurrence in meat products conducted so far have reported this occurrence to be high [ 26 , 27 , 28 , 29 ]. To the best of our knowledge, studies on STC occurrence in meat products have not been conducted yet, despite the EFSA CONTAM Panel recommendation to collect data on STC in food and feed using highly sensitive analytical methods with LOQ of less than 1.5 μg/kg.…”

“…This type of research has been encouraged because the data on STC incidence are limited, making it impossible to assess human and animal exposure [ 14 , 30 ]. The occurrence of CIT was examined in only one study of Croatian meat products, despite the fact that the strains of one of the main mycotoxin producers P. citrinum are often isolated from dry-cured meat products [ 27 , 31 ].…”

Occurrence of citrinin in the wheat grain cultivated in Kosovo and Albania during 2021

“…All isolates were identified according to Pitt and Hocking (2009) and Samson et al (2019). Molecular identification of mould isolates was performed to verify the results of traditional identification methods and was carried out in full accordance with Lešić et al (2021). The obtained sequences were aligned using Lasergene SeqManPro DNASTAR 13 (Madison, Wisconsin, USA).…”