First Survey on the presence of anisakid parasites in farmed European sea bass and gilthead sea bream produced and marketed in Sicily

The Scientific Opinion of EFSA (2010) outlines as the only fish free of health risks related to Anisakidae parasites the farmed salmon, if reared in floating cages or on-shore cages and fed by feed with no live parasites. Otherwise, the food operator shall verify, by means of procedures approved by the competent authority, that the fishery products do not represent a health risk regarding the presence of live parasites. This opinion was confirmed by several studies conducted on farmed salmon in Norway (Angot and Brasseur, 1993, Aquaculture, 118: 339-344; Lunestad, 2003, J Food Prot, 66:122-124). The absence of anisakid parasites in farmed fish has also been found for other marine fish species such as European sea bass (Dicentrarchus labrax) and gilthead sea bream (Sparus aurata) (Penalver et al., 2010 J Food Prot, 73:1332-1334). Currently there are low evidences on the prevalence of anisakid infestation in aquaculture fish produced and marketed in Italy. In this work, a sampling plan was carried out aimed at collecting different fish samples marketed in Sicily, to verify the presence and prevalence of anisakid parasites infestation. A total of 143 samples of sea bass and 110 samples of sea bream from Sicilian and Greek farms were examined: all the samples were of commercial size (over 200 g). The specimens were taken and stored refrigerated then transferred to the C.Re.N.A laboratories, where visual inspection of viscera and muscle was carried out. The negative samples were subjected to chloro-peptic digestion. The larvae found were subjected to morphological identification, through optical microscopy and molecular analysis by PCRRFLP method. The survey revealed the presence of two parasites belonging to the Anisakidae family, found inside the coelomatic cavity, only in a single sample of European sea bass from a single farm located in Greece, revealing a prevalence of infestation of 1.7%. The larvae were morphologically identified as belonging to the morphotype I of the genus Anisakis. Molecular investigations confirmed the larvae as Anisakis pegreffii species. No larvae were found in the samples of gilthead sea bream examined. The present work represents the first report on the presence of anisakid parasites in European sea bass. Our findings in farmed fish can be traced back to the aquaculture policies; however the prevalence of infestation in these productive realities remains very low. Furthermore, the results of this study suggest further investigations in order to have a comprehensive risk picture

Identification of contracaecum rudolphii (Nematoda: Anisakidae) in great cormorants phalacrocorax carbo sinensis (Blumenbach, 1978) from Southern Italy

Simple Summary: Four dead Phalacrocorax carbo sinensis specimens from Southern Italy coasts were examined for Contracaecum sp. detection. 181 Contracaecum sp. larvae and adults were found in the viscera of all the P. carbo sinensis examined. The PCR-RFLP analysis showed the presence of Contracaecum rudolphii A and B. A co-infestation of C. rudolphii A and B was found in P. carbo sinensis from Leporano Bay. This study provides a first report of the presence of Contracaecum sp. in P. carbo sinensis from Southern Italy. Abstract: In this study, four dead great cormorant Phalacrocorax carbo sinensis (Blumenbach, 1978) specimens, collected from the coasts and lakes of Southern Italy, were examined by necropsy for the detection of Contraceacum sp. The adults and larvae found were subjected to morphological analysis and molecular identification by PCR-RFLP. A total of 181 Contracaecum specimens were detected in all of the four great cormorants examined (prevalence = 100%), showing an intensity of infestation between nine and ninety-two. A co-infestation by adult and larval forms of Contracaecum rudolphii was found only in one of the great cormorants examined. Following molecular investigations, 48 specimens of C. rudolphii A and 38 specimens of C. rudolphii B were detected, revealing co-infestation solely for the great cormorant from Leporano Bay (Southern Italy). Our results showed an opposite ratio between C. rudolphii A and C. rudolphii B in Pantelleria and in Salso Lake (Southern Italy) compared to what was reported in the literature, probably due to migratory stopovers and the ecology of the infested fish species, confirming the role of Contracaecum nematodes as ecological tags of their hosts

Determination of Mycotoxins in Plant-Based Meat Alternatives (PBMAs) and Ingredients after Microwave Cooking

In this study, we investigate the role of microwave cooking in reducing mycotoxin contamination in plant-based food matrices, with a focus on veggie burgers (purchased and home-made) and their ingredients (soybean, potatoes, zucchini, carrots). Two different conditions were studied (Max–Min) that were 800 W for 60 s and 800 W for 90 s, respectively. The degradation patterns of aflatoxins (AFB1, AFB2, AFG1, AFG2), fumonisins (FB1, FB2, FB3), trichothecenes (T2, HT2, ZEA), and ochratoxin A (OTA) were studied. The extraction procedures were conducted with the QuEChERS extraction, and the analyses were conducted with liquid chromatography–tandem mass spectrometry (LC-MS/MS). Principal component analysis (PCA) showed that degradation under microwave cooking varies considerably across different food matrices and cooking conditions. This study provides valuable insights into the degradation of mycotoxins during microwave cooking and underscores the need for more research in this area to ensure food safety

PRESENCE OF ANISAKIS PEGREFFII IN FARMED SEA BASS ( DICENTRARCHUS LABRAX L.) COMMERCIALIZED IN SOUTHERN ITALY: A FIRST REPORT

We examined 151 European sea bass (Dicentrarchus labrax L.) samples from farms and fish markets of Sicily (Southern Italy) for Anisakidae larvae detection. All the samples were examined by visual inspection and modified chloro-peptic digestion. Two nematode larvae were found in the viscera of only one European sea bass sample from a farm located in Greece (FAO 37.3), giving a total prevalence of infestation of 0.7%. No other parasites were found after chloro-peptic digestion of the samples. The larvae were morphologically ascribed, at genus level, to morphotypes I and molecularly identified as Anisakis pegreffii. To the best if our knowledge, this is the first report on the presence of anisakid parasites in farmed European sea bass of Mediterranean Sea. Our findings suggest that the risk of exposure to Anisakidae nematodes in farmed European sea bass remains very low. However, further data on Mediterranean farms are needed to have a detailed risk analysis

QuEChERS LC\u2013MS/MS Screening Method for Mycotoxin Detection in Cereal Products and Spices

We developed and validated a screening method for mycotoxin analysis in cereal products and spices. Ultra-high-performance liquid chromatography coupled with tandem mass spectrometry (UHPLC–MS/MS) was used for the analysis. Dispersive solid-phase extractions (d-SPEs) were used for the extraction of samples. Ochratoxin A (OTA), zearalenone (ZEA), aflatoxins (AFLA; AFB1, AFB2, AFG1, AFG2), deoxynivalenol (DON), fumonisin (FUMO; FB1, FB2, FB3), T2, and HT2 were validated in maize. AFLA and DON were validated in black pepper. The method satisfies the requirements of Commission Regulation (EC) no. 401/2006 and (EC) no. 1881/2006. The screening target concentration (STC) was under maximum permitted levels (MLs) for all mycotoxins validated. The method’s performance was assessed by two different proficiencies and tested with 100 real samples

Evaluation of Anisakis presence in fish products of Sicily and survey on the efficiency of storage methods on the viability and allergy potential

The research aimed to evaluate the prevalence, geographical distribution and seasonal occurrence of Anisakidae nematodes infestation in fish products marketed in Sicily. A study on mercury detection was also conducted in order to evaluate a difference in the accumulation between parasitized and not parasitized fishes. We examined 1419 fish samples for Anisakidae larvae detection by visual inspection and digestion method. The collected larvae were identified at genus level by optical microscopy and at species level by molecular methods (PCR-RFLP and multiplex- PCR). A Real Time PCR was carried out for the assessment of DNA of Anisakidae presence in 23 transformed fish products. The method showed high sensitivity and specificity for the scope. The results obtained revealed the presence of Anisakidae larve in 463 fish samples (32,6%) and of Anisakidae DNA in 3 trasformed fish products (13%). All the larvae detected in transformed fish products were dead, with absence of motility until 24 h. Molecular analysis confirm Anisakis pegreffii as the prevalent species of Mediterranean Sea. Furthermore, the presence of A. simplex s.s., A. physeteris, A. pegreffii/A. simplex s.s. hybrid forms and Hysterothylacium fabri was verified with lower prevalence values. A significant correlation was found between Anisakidae infestation and presence of heavy metals (Hg) levels (p<0.05)

Aflatoxin M1 in cow, sheep, and donkey milk produced in Sicily, Southern Italy

Samples (n = 485) of raw (n = 394) or heat-treated (n = 91) milk of three different species (cow, n = 170; sheep, n = 133; donkey, n = 84), collected 2013–2016 in Western Sicily (Southern Italy), were analyzed for aflatoxin M1 (AFM1) by enzyme-linked immunosorbent assay (ELISA). Positive ELISA results were further analyzed by HPLC with fluorescence detection. Both methods had a detection limit for AFM1 in milk of 7 ng kg−1. ELISA yielded 12.9 and 5% positives in cows and sheep milk, respectively, all samples of donkey milk were negative. Levels of AFM1 were in most cases at 0.007–&lt; 0.05 μg kg−1, only two samples (sheep milk) slightly exceeded the European Union maximum level of 0.05 μg kg−1. Only 6% of the samples were positive for AFM1 in a concentration range of 0.008–0.15 μg kg−1. Only milk samples collected directly from farms were positive. Overall, the levels were much lower than previously reported for Southern Italy cow and sheep milk samples purchased in retail stores. The results of this work indicate a continuous improvement of the feeding techniques on dairy farms of Southern Italy, which is essential to ensure consumers’ protection

Seasonal trend of Anisakidae infestation in South Mediterranean bluefish

A total of 1104 fish samples from markets of Sicily were analysed for the detection and species identification of Anisakidae nematodes. The preliminary analysis of the fish samples showed the presence of 2459 larvae. All the fish species revealed different prevalence of infestation, with a maximum of 100% for Lepidopus caudatus and a minimum of 4.5% in Sardina pilchardus. The 80% of the larvae examined by PCR-RFLP analysis belonged to Anisakis pegreffii species. The seasonal infestation trend of Anisakis was evaluated in all the fish sample examined. The results of the seasonal infestation trend showed a marked connection with the ecological aspects of the fish species examined. As far as we know, this work report for the first time important ecological aspects of Lepidopus caudatus specimens of South Mediterranean. This work could be useful to plan a seasonal fishing strategy aimed at reducing the health risks related to Anisakis

Mercury accumulation in Mediterranean Fish and Cephalopods Species of Sicilian coasts: correlation between pollution and the presence of Anisakis parasites

<p>The aim of this study was to investigate mercury accumulation in some species, caught in Mediterranean Sea, in the period between May and December 2015, and to compare it to the presence of Anisakis parasites. The samples were examined by direct mercury analyzer (DMA-80) for their Hg levels. The metal concentration was compared to the presence or the absence of Anisakis parasites. Significant differences in Hg concentration in analysed samples were observed. The low-infested fishes contained 1–6 larvae of parasites whereas the high-infested one had 7–83 larvae.</p