Publications by authors named "Gaetano Cammilleri"

33 Publications

Nutritional and mineral composition of persimmon fruits ( L.) from Central and Southern Italy.

Nat Prod Res 2021 May 7:1-6. Epub 2021 May 7.

Science4life, Spin company University of Messina, Messina, Italy.

In this study, the nutritional and mineral composition of fruits from Apulia, Campania, Lazio, Sardinia and Sicily regions was evaluated. Dietary fiber, mineral, pectin, polyphenol, and protein contents were evaluated. Particularly high are the contents of the dietary fiber and pectins. The mineral elements profile was interesting due to its modest content of sodium and high potassium concentration. Protein amounts were in line with the quantities for this fruit. The total polyphenol content of the fruits analysed was very variable, interesting for the quantities found both total and gallic acid. The results obtained confirmed the nutritional value of this fruit even for special dietary regimens such as hypertension and heart diseases and the genuineness of its cultivation in Central-Southern Italy.
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http://dx.doi.org/10.1080/14786419.2021.1921768DOI Listing
May 2021

Acrylamide assessment of wheat bread incorporating chia seeds ( L.) by LC-MS/MS.

Food Addit Contam Part A Chem Anal Control Expo Risk Assess 2021 Mar 22;38(3):388-395. Epub 2021 Jan 22.

Istituto Zooprofilattico Sperimentale Della Sicilia, Palermo, Italy.

We examined the acrylamide content in samples of wheat bread with chia seeds added at different concentrations (2%, 5%, 7%, 10%) and cooked at predefined conditions (20 min at 200°C) by a validated LC-MS/M method after QuEChERS extraction. The acrylamide contents of the bread samples with added chia seeds were compared with control wheat bread samples. The highest acrylamide values were found in bread with 5% chia seeds, showing a mean value of 156.5 ± 115.4 µg/kg, followed by bread with 10% chia seeds (150.2 ± 103.8 µg/kg). About 6% of the bread samples with added chia seeds reached acrylamide levels above the benchmark level set by the EU Regulation. No significant differences in acrylamide values were found between control samples and bread with different percentages of chia seeds ( > .05). The results obtained provide a first report on the possible contribution of chia to the increase of acrylamide formation in bread.
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http://dx.doi.org/10.1080/19440049.2020.1853823DOI Listing
March 2021

Development of a Rapid and Eco-Friendly UHPLC Analytical Method for the Detection of Histamine in Fish Products.

Int J Environ Res Public Health 2020 10 13;17(20). Epub 2020 Oct 13.

Istituto Zooprofilattico Sperimentale della Sicilia 'A. Mirri', via Gino Marinuzzi, 3, 90129 Palermo, Italy.

We developed, validated, and confirmed with proficiency tests a fast ultra-high-performance liquid chromatography with diode array detector (UHPLC-DAD) method to determine histamine in fish and fishery products. The proposed method consists of two successive solid-liquid extractions: one with a dilute solution of perchloric acid (6%) and the second only with water. The instrumental analysis with UHPLC provides a very fast run time (only 6 min) with a retention time of approximately 4 min, a limit of quantification (LOQ) of 7.2 mg kg, a limit of detection (LOD) of 2.2 mg kg, a recovery around 100%, a relative standard deviation (RSD%) between 0.5 and 1.4, and an r of calibration curve equal to 0.9995. The method detected optimal values of the validation parameters and required a limited number of reagents in comparison to other methods reported in the literature. Furthermore, the method could detect histamine in a very short time compared with other methods. This method, in addition to being validated, precise, specific, and accurate, avoids wasting time, money, and resources, and limits the use of organic solvents.
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http://dx.doi.org/10.3390/ijerph17207453DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7602054PMC
October 2020

fish waste oil: Fatty acids composition and antibacterial activity.

PeerJ 2020 19;8:e9299. Epub 2020 Jun 19.

STEBICEF, University of Palermo, Palermo, Italy, Italy.

Background And Aims: Fish by-products are generally used to produce fishmeal or fertilizers, with fish oil as a by-product. Despite their importance, fish wastes are still poorly explored and characterized and more studies are needed to reveal their potentiality. The goal of the present study was to qualitatively characterize and investigate the antimicrobial effects of the fish oil extracted from waste samples and to evaluate the potential use of these compounds for treating pathogen infections.

Methods: waste samples were divided in two groups: heads and soft tissues. Fatty acids composition, and in particular the content in saturated (SAFAs), mono-unsaturated (MUFAs) and Polyunsaturated (PUFAs) fatty acids, was characterized through GC/MS Thermo Focus GC-DSQ II equipped with a ZB-5 fused silica capillary tubes column. The antimicrobial activity of the salmon waste oils was evaluated through the Minimum Inhibitory Concentration assay and the antibiotics contamination was determined by Liquid Chromatography with tandem Mass Spectrometry (LC-MS/MS) analysis. All experiments were done at least in triplicate.

Results: GC/MS analysis has shown the specific fatty acid composition of the salmon waste oils and their enrichment in MUFAs and PUFAs, with special reference to omega-3, -6, -7, -9 fatty acids. Furthermore, our study has highlighted the antimicrobial activity of the fish waste oil samples against two Gram+ and Gram- bacterial strains.

Conclusions: These data confirm that the fish waste is still quantitatively and qualitatively an important source of available biological properties that could be extracted and utilized representing an important strategy to counteract infective diseases in the context of the circular economy.
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http://dx.doi.org/10.7717/peerj.9299DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7307567PMC
June 2020

High hydroxycinnamic acids contents in fennel honey produced in Southern Italy.

Nat Prod Res 2020 Feb 5:1-6. Epub 2020 Feb 5.

Istituto Zooprofilattico Sperimentale della Sicilia "A. Mirri", Palermo, Italy.

A total of 122 honey samples ( ssp. Ligustica) collected from Southern Italy were examined for floral identification by melissopalynological examination and for polyphenols detection by an LC-ESI-Orbitrap -MS/MS method. The melissopalynological examination confirmed all the samples examined as fennel () unifloral variety. The analytical method carried out for polyphenols detection showed satisfactory linearity and recovery values, achieved during the validation of the method. Very high amounts of flavonols (kaempferol and quercetin) and hydroxycinnamic acids (caffeic acid, chlorogenic acid and ferulic acid), were found in all the samples examined. Among the hydroxycinnamic acids group, caffeic acid showed the highest mean contents (865.90 ± 67.07 µg/kg). The results of this work confirmed the high presence of phenolic acids with strong free radical-scavenging activity in fennel products such as honey, suggesting their use to reduce oxidative stress.
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http://dx.doi.org/10.1080/14786419.2020.1723090DOI Listing
February 2020

Validation of a Commercial Loop-Mediated Isothermal Amplification (LAMP) Assay for the Rapid Detection of spp. DNA in Processed Fish Products.

Foods 2020 Jan 16;9(1). Epub 2020 Jan 16.

Istituto Zooprofilattico Sperimentale della Sicilia, via Gino Marinuzzi 3, 90129 Palermo, Italy.

Parasites belonging to the genera are organisms of interest for human health because they are responsible for the Anisakiasis zoonosis, caused by the ingestion of raw or undercooked fish. Furthermore, several authors have reported this parasite to be a relevant inducer of acute or chronic allergic diseases. In this work, a rapid commercial system based on Loop-Mediated Isothermal Amplification (LAMP) was optimised and validated for the sensitive and rapid detection of spp. DNA in processed fish products. The specificity and sensitivity of the LAMP assay for processed fish samples experimentally infected with spp. larvae and DNA were determined. The LAMP system proposed in this study was able to give positive amplification for all the processed fish samples artificially contaminated with spp., giving sensitivity values equal to 100%. Specificity tests provided no amplification for the , , or genera and uninfected samples. The limit of detection (LOD) of the LAMP assay proposed was 10 times lower than the real-time PCR method compared. To the best of our knowledge, this is the first report regarding the application of the LAMP assay for the detection of spp. in processed fish products. The results obtained indicate that the LAMP assay validated in this work could be a reliable, easy-to-use, and convenient tool for the rapid detection of DNA in fish product inspection.
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http://dx.doi.org/10.3390/foods9010092DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7022600PMC
January 2020

Histamine in Fish Products Randomly Collected in Southern Italy: A 6-Year Study.

J Food Prot 2020 Jan 16:241-248. Epub 2020 Jan 16.

Istituto Zooprofilattico Sperimentale della Sicilia "A. Mirri," Via Gino Marinuzzi 3, 90129, Palermo, Italy (ORCID: https://orcid.org/0000-0001-9262-116X [F.G.G.]).

In total, 4,615 fresh and processed fish samples collected from 2010 to 2015 were analyzed for histamine by ultrahigh-performance liquid chromatography with diode array detection. Histamine levels were detected in 352 (7.6%) samples, with a maximum of 4,110 mg kg and mean values of 908.9 ± 1,226.79 and 344.01 ± 451.18 mg kg for fresh and processed fish samples, respectively. No histamine levels were found in canned tuna and smoked fish samples in contrast to most of the data reported in the literature. A low percentage (2.79%) of noncompliant samples was found. The highest mean values were found during 2011 and 2015 for fresh and processed fish samples, respectively, showing a significant ( < 0.05) difference between the sampling years. The histamine contents found in fresh fish samples were significantly higher ( < 0.05) than those of processed samples. Most of the positive samples came from street vendors, suggesting the need to improve inspection measures in these commercial categories to ensure fish product safety.
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http://dx.doi.org/10.4315/0362-028X.JFP-19-305DOI Listing
January 2020

Land Snails as a Valuable Source of Fatty Acids: A Multivariate Statistical Approach.

Foods 2019 Dec 12;8(12). Epub 2019 Dec 12.

Istituto Zooprofilattico Sperimentale della Sicilia, via Gino Marinuzzi 3, 90129 Palermo, Italy.

The fatty acid (FA) profile of wild , and land snail samples, collected in Sicily (Southern Italy), before and after heat treatment at +100 °C were examined by gas chromatography with a flame ionization detector (GC-FID). The results show a higher content of polyunsaturated fatty acids (PUFAs) in all of the examined raw snails samples, representing up to 48.10% of the total fatty acids contents, followed by monounsaturated fatty acids (MUFAs). The thermal processing of the snail samples examined determined an overall reduction of PUFA levels (8.13%, 7.75%, and 4.62% for , and samples, respectively) and a species-specific variation of saturated fatty acid (SFA) contents. Oleic acid remained the most abundant FA of all of the snails species examined, accounting for up to 29.95% of the total FA content. A relevant decrease of ɷ3/ɷ6 ratio was found only for samples. The principal component analysis (PCA) showed a separation of the snail samples in terms of species and heat treatment. The results of this work suggest land snails as a valuable source of MUFA and PUFA contents and boiling as appropriate treatment, according to the maintenance of healthy properties.
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http://dx.doi.org/10.3390/foods8120676DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6963810PMC
December 2019

Mercury Detection in Benthic and Pelagic Fish Collected from Western Sicily (Southern Italy).

Animals (Basel) 2019 Aug 22;9(9). Epub 2019 Aug 22.

Food Department, Istituto Zooprofilattico Sperimentale della Sicilia, via Gino Marinuzzi 3, 90129 Palermo, Italy.

In highly polluted water, fish can accumulate mercury up to a concentration of 10 mgKg. This has occurred on the eastern coasts of Sicily (Southern Italy), probably due to the intense industrial activity of this area. However, little is known about Hg accumulation in fish of the western Sicilian coasts. In this work, we examined the Hg accumulation of 108 fish samples belonging to 14 species collected from western Sicilian coasts using a direct mercury analyzer. The samples showed a mean mercury concentration of 0.165 ± 0.22 mg kg with a maximum in (1.72 mgKg), exceeding the limits provided by EC Reg. 1881/2006. The lowest Hg levels were found in samples (0.001 mgKg). A significant difference was found between the fish species examined ( < 0.05). The comparison between benthic and pelagic species did not show statistical differences ( < 0.05). Fish food constitutes the main route of Hg uptake for humans. Only four of the 130 samples examined reached a mercury concentration over the European limits. The comparative analysis of Hg pollution for benthic and pelagic species did not confirm a different trend in metal contamination.
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http://dx.doi.org/10.3390/ani9090594DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6769492PMC
August 2019

Toxic mineral elements in from Sicilian coasts (Southern Italy).

Nat Prod Res 2020 Jan 3;34(1):177-182. Epub 2019 Jul 3.

Istituto Zooprofilattico Sperimentale della Sicilia "A. Mirri", Palermo, Italy.

We assessed the relationship between V, Cr, Mn, Hg, As, Cd, Sn, Sb and Pb concentrations in samples from the coasts of Sicily and the expression of . Toxic mineral elements assessment was carried out by A.A. Spectrometry and ICP-MS. The expression was performed by q-PCR method. Low metals' levels were found in the mussel samples examined, in comparison with what was reported in literature. The highest mean values of toxic mineral elements were found in Gela (Cr 0.178 ± 0.03 mg/Kg, Mn 4.325 ± 0.012 mg/Kg, As 3.706 ± 0.009 mg/Kg, Sn 0.148 ± 0.014 mg/Kg, Sb 0.009 ± 0.004 mg/Kg e Pb 0.364 ± 0.01 mg/Kg). Significant levels of Hg were found in samples from Catania (0.014 ± 0.005 mg/Kg). Only vanadium and lead concentrations showed significant differences between sampling areas ( < 0.05). Molecular analysis verified a basal expression of and the absence of over-expression of , confirming the low mineral's concentrations found in the samples examined.
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http://dx.doi.org/10.1080/14786419.2019.1610963DOI Listing
January 2020

Seasonal trend of Anisakidae infestation in South Mediterranean bluefish.

Nat Prod Res 2020 Jan 20;34(1):158-161. Epub 2019 Feb 20.

Istituto Zooprofilattico Sperimentale della Sicilia "A. Mirri", Palermo, Italy.

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 and a minimum of 4.5% in The 80% of the larvae examined by PCR-RFLP analysis belonged to species. The seasonal infestation trend of 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 specimens of South Mediterranean. This work could be useful to plan a seasonal fishing strategy aimed at reducing the health risks related to .
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http://dx.doi.org/10.1080/14786419.2019.1573232DOI Listing
January 2020

Tetracycline Residues in Bovine Muscle and Liver Samples from Sicily (Southern Italy) by LC-MS/MS Method: A Six-Year Study.

Molecules 2019 Feb 15;24(4). Epub 2019 Feb 15.

Istituto Zooprofilattico Sperimentale della Sicilia "A. Mirri", via Gino Marinuzzi 3, 90129 Palermo, Italy.

We examined a total of 369 bovine liver and muscle samples for the detection of oxytetracycline (OTC), tetracycline (TC), chlortetracycline (CTC), and doxycycline (DOX) residues by implementation and validation of a LC-MS/MS method. The method showed good recovery values between 86% and 92% at three levels of concentrations. The linearity tests revealed r² > 0.996 for all the tetracyclines examined. Furthermore, the Youden test revealed that the method was robust. Only 14.4% of the samples showed OTC and TC residues in a concentration range of 10.4⁻40.2 µg kg. No CTC and DOX residues were found in all the samples analyzed. Liver samples showed the highest average values (31.5 ± 20.6 and 21.8 ± 18.9 for OTC and TC, respectively). The results showed a low incidence of TCs in all the samples examined, in comparison with other studies reported in the literature. A significant decrease in TC residues frequency was found from 2013 ( < 0.05). This work reports for the first time epidemiological data on the presence of TC residues in liver and muscle samples of cattle farmed in Sicily (Southern Italy). The very low incidence of TC residues indicates a continuous improvement in farming techniques in Southern Italy, which is essential to ensure consumers' protection.
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http://dx.doi.org/10.3390/molecules24040695DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6413177PMC
February 2019

Relationship between arsenic accumulation in tissues and hematological parameters in mullet caught in Faro Lake: a preliminary study.

Environ Sci Pollut Res Int 2019 Mar 2;26(9):8821-8827. Epub 2019 Feb 2.

Department of Veterinary Sciences, Polo Universitario Annunziata, University of Messina, 98168, Messina, Italy.

The authors investigated the arsenic (As) accumulation in different tissues (muscle, gill, liver, stomach, and intestine) and the possible correlation between tissue concentration and hematological parameters in mullet (Mugil cephalus Linnaeus, 1758) caught in Faro Lake (Messina, Sicily, Italy). On all fish, hematological analyses of blood samples, measurement of biometric indices, and the removal of the muscles, gills, liver, stomach, and intestine for the determination of arsenic concentration were performed. A hemogram was performed to find effects of arsenic concentration in tissues on hematological variables. One-way analysis of variance showed significant differences of arsenic concentration in different tissues, with higher values in the gill. The correlation between hematological parameters and tissue arsenic concentration showed a statistical significance for red blood cell (RBC), hemoglobin concentration (Hb), and hematocrit (Hct) with the liver As concentration. Biometric indices (weight, length, and fork length) showed a significant correlation with As concentration of the muscle and liver also. Our results indicate the role of some hematological parameters as biomarkers useful to monitoring anthropogenic load of arsenic in water and sediment, because variations of these parameters represent one of the effects that arsenic exposure can have on fish.
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http://dx.doi.org/10.1007/s11356-019-04343-7DOI Listing
March 2019

Residues of 165 pesticides in citrus fruits using LC-MS/MS: a study of the pesticides distribution from the peel to the pulp.

Nat Prod Res 2020 Jan 19;34(1):34-38. Epub 2019 Jan 19.

Istituto Zooprofilattico Sperimentale della Sicilia "A. Mirri", Palermo, Italy.

A sensitive LC-ESI-MS/MS method was developed for the determination of 165 pesticides in 50 citrus fruit samples collected in Sicily. Moreover, an evaluation of pesticides levels in the citrus layers (peel, albedo, and pulp) was carried out. The method presented acceptable trueness, precision, and linearity with LOQ of 5 μg/kg. The results obtained showed a high frequency of fungicides class pesticides in all the citrus samples examined (>95%) with the highest concentrations in the peel (4468 µg/Kg). A significant difference of concentrations was found between the layers of the citrus fruits analysed (p < 0.05). In particular, the peel and albedo present higher pesticides significantly higher than the pulp. Our findings confirming the widespread use of these substances in citrus cultivation and suggesting the importance of pesticides analysis in all the citrus fruit layers separately, considering the different interactions between the physicochemical characteristics of the matrices and the pesticides.
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http://dx.doi.org/10.1080/14786419.2018.1561682DOI Listing
January 2020

Prevalence of anisakid parasites in fish collected from Apulia region (Italy) and quantification of nematode larvae in flesh.

Int J Food Microbiol 2019 Mar 26;292:159-170. Epub 2018 Dec 26.

Istituto Zooprofilattico Sperimentale della Sicilia, National Reference Centre for Anisakiasis (C.Re.N.A.), Palermo, Italy.

Anisakis spp. and Hysterothylacium spp. are nematodes that commonly parasitize several fish species. Nematode larvae can be recovered in coelomic cavity and viscera, but also in flesh and have an important economic and public health impact. A total of 1144 subjects of wild teleosts, 340 samples of cephalopods and 128 specimens of farmed fish collected from Apulia region were analysed for anisakid larvae detection by visual inspection of coelomic cavity and viscera and by digestion of the flesh. No nematode larvae were found in farmed fish and cephalopod molluscs. All examined wild-caught fish species were parasitized, except for 5 species for each of which only a few subjects belonging to the same batch were sampled, therefore the results are just indicative. A total of 6153 larvae were isolated; among these, 271 larvae were found in the muscular portion. Larvae were identified by morphological method as belonging to the genera Anisakis (97.2%) (type I and type II) and Hysterothylacium (2.8%). Both nematodes could be found in all fish species, except for round sardinella (Sardinella aurita), infected only by Hysterothylacium spp. and for Mediterranean scaldfish (Arnoglossus laterna), little tunny (Euthynnus alleteratus) and chub mackerel (Scomber japonicus) infected only with Anisakis spp.. A sample of 185 larvae was sent to the National Reference Centre for Anisakiasis (C.Re.N.A.) of Sicily for identification at the species level: 180 larvae belonged to the species A. pegreffii and 2 larvae to A. physeteris. The remaining 3 larvae were identified at genus level as Hysterothylacium. Statistical indices such as prevalence, mean intensity and mean abundance were calculated. Chub mackerel (S. japonicus) was the species with the highest prevalence and mean intensity. Moreover, the average and the median values of larvae per 100 g of edible part for each fish species were determined to estimate the consumer exposure to Anisakis spp.. The obtained values were then recalculated by referring to the edible part of all specimens (infected and non-infected) forming a single parasitized batch, getting more realistic and objective data useful for risk assessment. Our results indicate that the consumption of raw or undercooked wild fish caught off Apulian coasts could result in the acquisition of anisakiasis; on the contrary, farmed fish and cephalopods appear to be safer for the consumer.
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http://dx.doi.org/10.1016/j.ijfoodmicro.2018.12.025DOI Listing
March 2019

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

Mycotoxin Res 2019 Feb 13;35(1):47-53. Epub 2018 Sep 13.

Istituto Zooprofilattico Sperimentale della Sicilia "A. Mirri", via Gino Marinuzzi 3, 90129, Palermo, 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 M (AFM) by enzyme-linked immunosorbent assay (ELISA). Positive ELISA results were further analyzed by HPLC with fluorescence detection. Both methods had a detection limit for AFM in milk of 7 ng kg. ELISA yielded 12.9 and 5% positives in cows and sheep milk, respectively, all samples of donkey milk were negative. Levels of AFM were in most cases at 0.007-< 0.05 μg kg, only two samples (sheep milk) slightly exceeded the European Union maximum level of 0.05 μg kg. Only 6% of the samples were positive for AFM in a concentration range of 0.008-0.15 μg kg. 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.
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http://dx.doi.org/10.1007/s12550-018-0329-yDOI Listing
February 2019

Presence of Anisakis pegreffii in farmed sea bass (Dicentrarchus labrax L.) commercialized in Southern Italy: A first report.

Vet Parasitol 2018 Aug 28;259:13-16. Epub 2018 Jun 28.

Istituto Zooprofilattico Sperimentale della Sicilia "A. Mirri", via Gino Marinuzzi 3, 90129 Palermo, Italy.

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.
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http://dx.doi.org/10.1016/j.vetpar.2018.06.021DOI Listing
August 2018

Molecular Identification of Hysterothylacium spp. In Fishes From the Southern Mediterranean Sea (Southern Italy).

J Parasitol 2018 08 16;104(4):398-406. Epub 2018 May 16.

Istituto Zooprofilattico Sperimentale della Sicilia, Via Gino Marinuzzi 3, 90129 Palermo, Italy.

Samples obtained from 11 teleost fish species collected in waters off of the Mediterranean coasts of Sicily, Italy, were examined for the presence of Hysterothylacium spp. larvae. In total, 3,017 fish samples were examined, and the larvae recovered were used in subsequent phylogenetic studies. Fifty-eight raphidascaridid parasitic nematodes were found in the examined fish, with prevalence values ranging from 0.2% in anchovies to 60% in forkbeard samples. Twenty-seven parasites were identified as Hysterothylacium fabri and Hysterothylacium aduncum by sequencing the following regions: the nuclear ribosomal internal transcribed spacer ( ITS) and the mitochondrial cytochrome c oxidase subunit II ( COXII). Two other larvae collected from Lophius piscatorius had very low identity scores, with ITS and COXII sequences reported in GenBank and high genetic distances (AY603539). Pairwise comparisons between the ITS region of the H. fabri isolated from fishes from Sicilian coastal waters and those isolated from the Mediterranean Sea, Turkey (KC852206), revealed genetic differences ranging from 0.015 to 0.018. Our H. aduncum samples had very low genetic differences to H. aduncum from the Adriatic Sea (KP979763, 0.00-0.003). The concatenated phylogenetic examination of the ITS- COXII sequences by using maximum likelihood analyses indicated 3 distinct clades supported by high bootstrap values. Further molecular identification and detailed morphological analyses are needed to clarify these results and confirm the diversity and relationships within Hysterothylacium.
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http://dx.doi.org/10.1645/16-60DOI Listing
August 2018

Corticosteroids in liver and urine in Sicilian cattle by a LC-MS/MS method.

Food Addit Contam Part B Surveill 2018 Sep 22;11(3):201-207. Epub 2018 May 22.

a Area Chimica e Tecnologie Alimentari , Istituto Zooprofilattico Sperimentale della Sicilia "A. Mirri" , Palermo , Italy.

The presence of corticosteroid residues was assessed in urine and liver samples from livestock of Sicily. A total of 630 bovine samples were collected from farms and slaughterhouses. The samples were analysed using solid-phase extraction (SPE) coupled with ultra-performance liquid chromatography tandem mass spectrometry (UHPLC-MS/MS). All the corticosteroids found were under the maximum residue limit imposed by Commission Regulation (EC) 37/2010. About 4% of liver samples showed dexamethasone levels above the limit of detection (LOD), with a mean of 1.5 ± 0.2 µg kg. Betamethasone was found only in seven liver samples, with a mean of 1.6 ± 0.1 µg kg. Furthermore, prednisolone and prednisone were found only in urine and liver samples from slaughterhouse, probably related to the high rate of stress for bovines. These results suggest good control practices adopted by Sicilian farms, able to ensure the quality of food products.
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http://dx.doi.org/10.1080/19393210.2018.1470578DOI Listing
September 2018

Toxic metal levels in cocoa powder and chocolate by ICP-MS method after microwave-assisted digestion.

Food Chem 2018 Apr 15;245:1163-1168. Epub 2017 Nov 15.

Istituto Zooprofilattico Sperimentale Della Sicilia "A. Mirri", Via Gino Marinuzzi 3, 90129 Palermo, Italy.

The Commission Regulation (EC) Regulation N. 488/2014, established the concentration limits for cadmium in specific products based on cocoa and chocolate products as from January 2019. Based on this information there is a need to determine ultratrace levels of elements that might be presents in cocoa and chocolate products. In this work, the concentrations of Arsenic, Antimony, Cadmium, Chromium, Lead, Selenium and Vanadium were evaluated in cocoa powder and chocolate by the validation of an ICP-MS method. Good selectivity/specificity, recovery, repeatability and within-laboratory reproducibility, LOD, LOQ, range of linearity, standard measurement uncertainty parameters for method validation were achieved, in accordance with Commission Regulation. The cocoa powder revealed the maximum metal concentrations of 0.303 ± 0.035 mg/kg for cadmium, 1.228 ± 0.146 mg/kg for lead and 0.094 ± 0.013 mg/kg for arsenic. A significant difference was found between cocoa powder and chocolate samples (p < .05).
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http://dx.doi.org/10.1016/j.foodchem.2017.11.052DOI Listing
April 2018

Validation of a commercial kit aimed to the detection of pathogenic anisakid nematodes in fish products.

Int J Food Microbiol 2017 Sep 15;257:75-79. Epub 2017 Jun 15.

Department of Public Health and Infectious Diseases, Sapienza University of Rome, Italy. Electronic address:

Anisakids are parasitic nematodes responsible for a zoonosis that occurs following the ingestion of fish and fish products infected with larvae belonging to the genera Anisakis and Pseudoterranova. Rarely Contracaecum is found in association with gastric/intestinal illness, while Hysterothylacium is commonly considered not pathogenic. Although Real Time PCR assays have been recently used with the aim to detect and quantify these parasites in food products, methods applied did not undergo through extensive validation process, a feature highly desirable or mandatory in the case of testing laboratories accredited for the ISO EN 17025:2005. Here, a comprehensive study has been performed to validate a commercial kit based on multiplex real time PCR for the qualitative detection of Anisakis and Pseudoterranova. Inclusivity/exclusivity trials were carried out on DNA from species of the genera Anisakis, Pseudoterranova, Contracaecum, Hysterothylacium and Ascaris, on fish intentionally contaminated with Anisakis spp. and Pseudoterranova spp. and on marine organisms as fish, crustacean and squid to test the commercial kit on a large sample. The assay gave positive amplification for several Anisakis and Pseudoterranova species, while providing no signal for the members of the remaining genera. Each sample was correctly assigned either to Anisakis or Pseudoterranova, thus indicating that no cross-reaction occurred. The LOD was determined using two independent standard curves. Robustness was assayed by using two different thermocyclers in three distinct laboratories with different operators. The establishment of a validation dossier will permit the use of the commercial kit for the detection of Anisakis and Pseudoterranova DNA in fish and fish products intended for human consumption by public or private laboratories, following the requirements regarding the quality assurance processes described in the ISO EN 17025:2005.
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http://dx.doi.org/10.1016/j.ijfoodmicro.2017.06.011DOI Listing
September 2017

Survey on the presence of non-dioxine-like PCBs (NDL-PCBs) in loggerhead turtles (Caretta caretta) stranded in south Mediterranean coasts (Sicily, Southern Italy).

Environ Toxicol Chem 2017 11 26;36(11):2997-3002. Epub 2017 Jul 26.

Istituto Zooprofilattico Sperimentale della Sicilia "A. Mirri,", Palermo, Italy.

A total of 71 loggerhead turtles (Caretta caretta) stranded along the coasts of Sicily (Southern Italy) were examined for non-dioxine like polychlorinated biphenyl (NDL-PCB) levels in muscle and adipose tissue by a gas chromatography-mass spectrometry/mass spectrometry (GC-MS/MS) method. The results revealed 6 high-indicator congener (∑ PCB ) levels in 45% of the loggerhead turtles examined, with mean values of 980.39 ± 2508.39 ng/g wet weight in adipose tissue and 102.53 ± 238.58 ng/g wet weight in muscle tissue. The hexachloro and heptachloro PCB congeners were the most abundant in both the sample types. The highest NDL-PCB levels were reached in an adipose tissue sample of a loggerhead turtle of 80 kg stranded along the coasts of Termini Imerese (14 183.85 ng/g wet wt). No significant correlation was found between modified Fulton's K values of the loggerhead turtles and PCB contents (S = 47 151, p > 0.05). Furthermore, no significant differences were found between sexes (W = 365, p >  0.05). The PCB levels found in the present study were much higher than those found in the literature. The present study is the first report on the existence of NDL-PCBs in loggerhead turtles stranded in Sicilian coasts confirming C. caretta as a valuable indicator of contaminant exposure in the marine environment because of their specific biological and ecological characteristics. Environ Toxicol Chem 2017;36:2997-3002. © 2017 SETAC.
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http://dx.doi.org/10.1002/etc.3866DOI Listing
November 2017

Mercury in fish products: what's the best for consumers between bluefin tuna and yellowfin tuna?

Nat Prod Res 2018 Feb 31;32(4):457-462. Epub 2017 Mar 31.

a Instituto Zooprofilattico Sperimentale della Sicilia , Palermo , Italy.

A total of 205 bluefin and yellowfin tuna samples were examined for mercury detection in order to verify possible differences and have a detailed risk assessment of the two tuna species. The results showed significant higher mercury concentration in muscle tissue of bluefin tuna respect yellowfin tuna (p < 0.001) with mean concentration of 0.84 mg/kg and maximum value of 1.94 mg/kg. These differences can be due the different biological and ecological aspects of the two tuna species and to different oceanographic aspects between Atlantic Ocean and Mediterranean sea. The results obtained in this study suggest an advisable containment of the sources of pollution and further studies on the closed-loop farming of bluefin tuna, in order to ensure the product safety.
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http://dx.doi.org/10.1080/14786419.2017.1309538DOI Listing
February 2018

Determination of Ten Corticosteroids in Illegal Cosmetic Products by a Simple, Rapid, and High-Performance LC-MS/MS Method.

Int J Anal Chem 2017 14;2017:3531649. Epub 2017 Feb 14.

Istituto Zooprofilattico Sperimentale della Sicilia "A. Mirri", Via Gino Marinuzzi 3, 90129 Palermo, Italy.

The aim of our present work was the development of a rapid high-performance liquid chromatography method with electrospray ionization and tandem mass spectrometry detection (LC-ESI-MS/MS) for the determination of several corticosteroids in cosmetic products. Corticosteroids are suspected to be illegally added in cosmetic preparations in order to enhance the curative effect against some skin diseases. Sample preparation step consists in a single extraction with acetonitrile followed by centrifugation and filtration. The compounds were separated by reversed-phase chromatography with water and acetonitrile (both with 0.1% formic acid) gradient elution and detected by ESI-MS positive and negative ionization mode. The method was validated at the validation level of 0.1 mg kg. Linearity was studied in the 5-250 g L range and linear coefficients () were all over 0.99. The accuracy and precision of the method were satisfactory. The LOD ranged from 0.085 to 0.109 mg kg and the LOQ from 0.102 to 0.121 mg kg. Mean recoveries for all the analytes were within the range 91.9-99.2%. The developed method is sensitive and useful for detection, quantification, and confirmation of these corticosteroids in cosmetic preparations and can be applied in the analysis of the suspected samples under investigation.
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http://dx.doi.org/10.1155/2017/3531649DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5331168PMC
February 2017

Monitoring of neonicotinoid pesticides in beekeeping.

Nat Prod Res 2017 Jun 13;31(11):1258-1262. Epub 2016 Oct 13.

c Istituto Zooprofilattico Sperimentale della Sicilia "A. Mirri" via Gino Marinuzzi , Palermo , Italy.

The decline of pollinating species is correlated to the extensive use of neonicotinoids against pest insects for crop protection. In this study, the concentrations of neonicotinoid insecticides were determined in honeybees, honeycomb and honey samples, collected in Spring 2015 (blooming period) from different areas in Sicily (IT), to carry out an evaluation of bees products' safety and an overview of neonicotinoid contamination in beekeeping. The results obtained showed only the presence of clothianidin in bee samples and these concentrations don't represent a risk for bees' vitality and safety. The absence of residue in all honey samples, instead, showed the quality of bee products.
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http://dx.doi.org/10.1080/14786419.2016.1236101DOI Listing
June 2017

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

Nat Prod Res 2017 May 12;31(10):1156-1162. Epub 2016 Sep 12.

a Dipartimento di Chimica degli Alimenti , Istituto Zooprofilattico Sperimentale della Sicilia " A. Mirri" via Gino Marinuzzi , Palermo , Italy.

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.
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http://dx.doi.org/10.1080/14786419.2016.1230119DOI Listing
May 2017

Survey on the presence of A. simplex s.s. and A. pegreffii hybrid forms in Central-Western Mediterranean Sea.

Parasitol Int 2016 Dec 25;65(6 Pt A):696-701. Epub 2016 Aug 25.

Istituto Zooprofilattico Sperimentale della Sicilia, Via Gino Marinuzzi 3, Palermo, Italy.

In this study, 1029 fish and cephalopod samples came from Central-Western Mediterranean (FAO 37.1.1 and FAO 37.1.3) were analysed for Anisakidae larvae research with the aim to identify possible hybridisations between Anisakis pegreffii and Anisakis simplex s.s. species. A total of 1765 larvae were detected, with prevalence values between 8.1% and 100%. The morphologic analysis revealed characters attributable to morphotype I of Anisakis in 98.5% of the examined larvae, while 1.5% belonged to the morphotype II. PCR-based Restriction Fragment Length Polymorphism (PCR-RFLP) analysis of the entire ITS region (ITS1, 5.8S and ITS2) of nuclear ribosomal DNA (rDNA) was performed with HinfI and HhaI restriction enzymes. The majority of the larvae examined by PCR-RFLP were identified as A. pegreffii (71%), with a prevalence on horse mackerel from FAO 37.1.3, while 10% were identified as A. simplex s.s., 2% as A. physeteris and 17% as A. pegreffii×A. simplex s.s. hybrid genotype. The sequence analysis confirmed the hybridisation in the 85% of the larvae recognised as hybrid forms by PCR- RFLP, suggesting this form as the product of natural interspecific recombination due to the presence of sympatry areas. The presence of hybrid forms were mostly found in fish samples from FAO subzone 37.1.1. This is the first report of A. pegreffii x A. simplex s.s. hybrid genotype in fishes caught off the coasts of Sicily (Southern Italy). Finally, this study provided substantial information about the geographical distribution of Anisakidae family in Central-Western Mediterranean Sea.
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http://dx.doi.org/10.1016/j.parint.2016.08.004DOI Listing
December 2016

Are mussels able to distinguish underwater sounds? Assessment of the reactions of Mytilus galloprovincialis after exposure to lab-generated acoustic signals.

Comp Biochem Physiol A Mol Integr Physiol 2016 11 29;201:61-70. Epub 2016 Jun 29.

Istituto per l'Ambiente Marino Costiero U.O. di Capo Granitola, Consiglio Nazionale delle Ricerche, Via del mare no. 3, 91021 Torretta Granitola, TP, Italy. Electronic address:

This study examined the effects of lab-generated acoustic signals on the behaviour and biochemistry of Mediterranean mussels (Mytilus galloprovincialis). The experiment was carried out in a tank equipped with a video-recording system using six groups of five mussels exposed to five acoustic treatments (each treatment was replicated three times) for 30min. The acoustic signals, with a maximum sound pressure level of 150dB rms re 1μPa, differed in frequency range as follows: low (0.1-5kHz), mid-low (5-10kHz), mid (10-20kHz), mid-high (20-40kHz) and high (40-60kHz). The exposure to sweeps did not produce any significant changes in the mussels' behaviour. Conversely, the specimens exposed to the low frequency band treatment showed significantly higher values of the following biochemical stress parameters measured in their plasma and tissues: glucose, total proteins, total haemocyte number (THC), heat shock protein 70 (Hsp70) expression, and Acetylcholinesterase (AChE) activity. The responses observed in the mussels exposed to low frequency sweeps enable us to suppose a biological and ecological role for this sound, which contains the main frequencies produced by both shipping traffic and the acoustic emissions of fish.
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http://dx.doi.org/10.1016/j.cbpa.2016.06.029DOI Listing
November 2016

Fatty acids determination in Bronte pistachios by gas chromatographic method.

Nat Prod Res 2016 Oct 4;30(20):2378-82. Epub 2016 Jun 4.

a Food Department , Istituto Zooprofilattico Sperimentale della Sicilia , Palermo , Italy.

A gas chromatographic with flame ionization detector (GC-MS FID) method for the identification and quantification of fatty acids based on the extraction of lipids and derivatisation of free acids to form methyl esters was developed and validated. The proposed method was evaluated to a number of standard FAs, and Bronte pistachios samples were used for that purpose and to demonstrate the applicability of the proposed method. In this regard, repeatability, mean and standard deviation of the analytical procedure were calculated. The results obtained have demonstrated oleic acid as the main component of Bronte pistachios (72.2%) followed by linoleic acid (13.4%) and showed some differences in composition with respect to Tunisian, Turkish and Iranian pistachios.
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http://dx.doi.org/10.1080/14786419.2016.1180599DOI Listing
October 2016

Hair Mercury Levels Detection in Fishermen from Sicily (Italy) by ICP-MS Method after Microwave-Assisted Digestion.

Bioinorg Chem Appl 2016 5;2016:5408014. Epub 2016 Apr 5.

Istituto Zooprofilattico Sperimentale della Sicilia "A. Mirri", Via Gino Marinuzzi 3, 90129 Palermo, Italy.

A number of ninety-six hair samples from Sicilian fishermen were examined for total mercury detection by an Inductively Coupled Plasma Mass Spectrometry (ICP-MS) method. The mercury levels obtained were compared with mercury levels of 96 hair samples from a control group, in order to assess potential exposure to heavy metals of Sicilian fishermen due to fish consumption and closeness to industrial activities. Furthermore, the mercury levels obtained from hair samples were sorted by sampling area in order to verify the possible risks linked to the different locations. The overall mean concentration in the hair of the population of fishermen was 6.45 ± 7.03 μg g(-1), with a highest value in a fisherman of Sciacca (16.48 μg g(-1)). Hair mercury concentration in fishermen group was significantly higher than in control group (p < 0.01). There was no significant difference in hair total mercury concentrations between sampling areas (p > 0.05). The results of this study indicate a greater risk of exposure to mercury in Sicilian fishermen, in comparison to the control population, due to the high consumption of fish and the close relationship with sources of exposure (ports, dumps, etc.).
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http://dx.doi.org/10.1155/2016/5408014DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4835620PMC
April 2016
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