Production of a new chitinase from Nocardiopsis halophila TN-X8 utilizing bio-waste from the blue swimming crab: enzyme characterization and immobilization.

In accordance with the framework of the Circular Blue Bioeconomy in the Mediterranean region, the objective of this study was to evaluate the biotransformation of blue swimming crab (Portunus segnis) residues obtained from the port of Sfax by an extracellular chitinase produced by Nocardiopsis halophila strain TN-X8 isolated from Chott El Jerid (Tozeur, Tunisia). From the analysis of multiple extremophilic Actinomycetota, it was determined that strain TN-X8 exclusively utilized 60 g/L of raw blue swimming crab as its carbon and energy source, achieving a chitinase activity of approximately 950 U/mL following a 6-day incubation period at 40 °C. Pure chitinase, designated as ChiA-Nh30, was obtained after heat treatment, followed by ammonium sulfate fractionation and Sephacryl® S-200 column chromatography. The maximum ChiA-Nh30 activity was observed at pH 3 and 75 °C. Interestingly, compared with cyclohexamidine, ChiA-Nh30 showed a good antifungal effect against four pathogenic fungi. Furthermore, when using colloidal chitin as substrate, ChiA-Nh30 demonstrated a higher degree of catalytic efficiency than the commercially available Chitodextrinase®. In addition, ChiA-Nh30 could be immobilized by applying encapsulation and encapsulation-adsorption techniques. The kaolin and charcoal used acted as excellent binders, resulting in improved ChiA-Nh30 stability. For the immobilized ChiA-Nh30, the yield of N-acetyl-D-glucosamine monomers released from 20% (w/v) blue swimming crab residues increased by 3.1 (kaolin) and 2.65 (charcoal) times, respectively.

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

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.

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.

Heavy Metals and PAHs in Meat, Milk, and Seafood From Augusta Area (Southern Italy): Contamination Levels, Dietary Intake, and Human Exposure Assessment.

Heavy metals and PAHs were measured in animal foodstuffs from Augusta-Melilli-Priolo area in order to evaluate the potential human health risk associated to their consumption. All heavy metals were detected in seafood products while most of them were 1 for baby, children and teenagers, indicating a non-carcinogenic risk for these age categories by seafood ingestion. The CRAs overcame 1*10-5 for almost age categories (except "baby") and for elderly, by seafood and beef ingestions respectively. Moreover, the MOE for PAHs showed a certain cancer risk for "baby" related to cow milk ingestion.

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

Parasites belonging to the Anisakis 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 Anisakis spp. DNA in processed fish products. The specificity and sensitivity of the LAMP assay for processed fish samples experimentally infected with Anisakis 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 Anisakis spp., giving sensitivity values equal to 100%. Specificity tests provided no amplification for the Contracaecum, Pseudoterranova, or Hysterothylacium genera and uninfected samples. The limit of detection (LOD) of the LAMP assay proposed was 102 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 Anisakis 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 Anisakis DNA in fish product inspection.

Effects of different ozonized slurry-ice treatments and superchilling storage (-1°C) on microbial spoilage of two important pelagic fish species.

Combining different preservative treatments for improving quality and safety of fishery products increasingly receives global research attention. Consistent with this pursuit, the current research was undertaken to determine the effects of different ozonized slurry-ice treatments and superchilling (-1°C) storage on microbial spoilage of European anchovy (Eugraulis encrasicolus) and sardine (Sardina pilchardus), which are two commercially important pelagic fish species. After the catch (within <5 hr) and at defined scheduled storage times, ozone has been discharged once on sardine (herein referred to as "One-T") and repeatedly/sequentially on European anchovy (herein referred to as "Seq-T"). Microbiological analyses enumerated total viable count (TVC), Bacillus spp., Enterobacteriaceae, Lactobacillus spp., Moraxella spp., Shewanella spp., Listeria monocytogenes and Pseudomonas spp. Independent of potential antimicrobial effects of ozone during superchilling storage, no Listeria spp., Shewanella spp., Moraxella spp., and Bacillus spp. were found in all processed samples. While Enterobacteriaceae and Lactobacillus spp. were detected at below 1 log cfu/g, both TVC and Pseudomonas spp. proliferated at different rates throughout superchilling storage. The repeated ozone-treated ("Seq-T") showed lower TVC and Pseudomonas spp. values compared with one-time treated ("One-T") slurry-iced and control samples. Thus, combined slurry-ice and superchilling storage at Seq-T produced improved antimicrobial activity over One-T application. Largely, ozonized slurry-ice outcomes/results appear promising thanks to superchilling storage.

Risk factors for the persistence of bovine brucellosis in Sicily from 2008 to 2010.

Bovine brucellosis is a worldwide zoonotic disease, responsible for economic and reproductive losses in farmed animals. Despite the application of control and eradication measures, after several years the brucellosis situation still remains problematic in Sicily, one of the regions with the highest prevalence of infection in Italy. The objective of this paper is to evaluate the possible risk factors influencing the presence of bovine brucellosis in Sicily. Data on existing cattle herds in Sicily and on their structure in 2008, 2009 and 2010 were extracted from the national database (NDb) of holdings and animals. Several possible brucellosis risk factors were considered, in particular herd-related factors and the performances of the control activities carried out by the veterinary services and recorded into national animal health information system (SANAN). Statistical analysis (χ(2) and Mann-Whitney tests) and spatial cluster analysis using a simple spatial Bernoulli model were performed. The multivariable logistic regression was also performed using SPSS for Windows. The spatial distribution of infected herds shows two main clusters of infection, in Messina and in Siracusa provinces. The results of the statistical analysis performed show that the infection is associated to the largest herds (P<0.0001) and to beef and mixed herds (P<0.0001). A significant association can be observed between the infected status in 2010 and presence of positive animals in 2008 or in 2009 (P<0.0001). This article shows the importance of detailed and reliable epidemiological data through information systems, allowing a continuous monitoring of the health status of animal populations and also the evaluation and re-planning of veterinary activities.

Spatial distribution of brucellosis in sheep and goats in Sicily from 2001 to 2005.

Spatial analysis is making an increasingly important contribution to surveillance measures due to its ability to enable immediate visualization of information on the phenomenon studied. The authors describe the spatial distribution of prevalence and incidence of brucellosis in small ruminants in Sicily between 2001 and 2005. The study was conducted by integrating geographic information systems (GIS) technology (MapInfo Professional 7.0) with SaTScan software to perform an epidemiological analysis of the municipalities and to locate problem areas. A comparison between the thematic maps produced for brucellosis in small ruminants on the basis of prevalence and incidence data for each individual year has shown that in terms of prevalence, the area identified as the secondary cluster in 2001 became the primary cluster from 2002 onwards whereas, in terms of incidence, the distribution of the clusters was irregular throughout the entire region during the years studied.