Levels of Essential and Trace Elements in Mozzarella Available on the Slovak Market with the Estimation of Consumer Exposure.

The aim of this study was to determinate the content of some elements in a specific dairy product, mozzarella, in a particular area of western Slovakia and to evaluate the estimation of the risk to the consumers based on the contribution to the provisional tolerable weekly intake. The consumption of mozzarella can contribute to the intake of important elements in the diet, such as calcium and magnesium, along with others. The contents of some toxic and trace elements were low and have not exceeded the permitted limit. In addition, the contribution to PTWI was found to be very low, which means that the consumption of mozzarella possesses no risk to humans. It is concluded that the data obtained in this study can help as a valuable addition to methodological and scientific material in the field of food safety of dairy products and their positive impact on human health.

Food safety culture in food companies: evaluation of the perception of food safety culture in three Tuscan food companies.

The Commission Regulation (EU) No. 2021/382 (European Commission, 2021), amending the Regulation (EC) No. 852/2004 (European Commission, 2004), introduced the obligation for companies to establish and maintain a food safety culture (FSC). The methodology to evaluate, implement, and enhance the level of FSC is up to the individual companies. This study aimed to investigate the perception of FSC among the employees of 3 Tuscan medium-sized enterprises in the food sector, producing cured meat (A), dairy products (B), and frozen fish products (C). The survey was conducted through the development and administration of a questionnaire based on a 5 points Likert scale, referring to different aspects of FSC, organized in 6 sections with 5-6 statements each and subjected to a percentage of employees between 76 and 85%, classified also by the length of service (≤3 and >3 years). For all the companies, the minimum median and mode value for scores obtained by the different sections was 4, and the minimum median and mode value for the single statement was 3 (A, B; except for a bimodal value 2-4) and 4 (C). The section awareness and perception of risk showed the highest mean scores in all companies. As for the length of service, senior employees gave lower scores than junior ones in all sections in B and 3 sections in C. Overall, the results of the questionnaires showed a good perception of FSC, even though it was possible to identify some partial weaknesses.

Effects of various logistics packaging on the quality and microbial variation of bigeye tuna (Thunnus obesus).

Bigeye tuna (Thunnus obesus) is an economically valuable ocean fish species. It is susceptible to contamination during storage and transportation. Having proper transportation packaging and stable temperature during transportation are critical to prevent quality deterioration. However, the influence of packaging on retaining freshness in transit remains unknown. Here, the impact of different transportation packaging on the quality and microbiological variation of bigeye tuna during the logistics process was investigated by measuring physical-chemical indexes and microbial diversity. It turned out that aluminum foil paper (AFP) group had minimum temperature fluctuation, exhibited preferable water retaining capacity and color protection effect. AFP packaging could efficiently prevent TVB-N increase and microbial growth. After 40 h, the TVB-N value was 21.28 mg/100 g and microbial total plate count was 3.53 lg CFU/g, which was within the acceptable range. Temperature fluctuations and packaging materials had a major effect on the microbial community structure of bigeye tuna. Chitinophagaceae, Acinetobacter, and Knoellia were dominant in the AFP group, while Pseudomonas, Acinetobacter, and Macrococcus were dominant in the expanded polystyrene foam (EPSF) and European logistics (EUL) groups. AFP packaging could effectively slow down the growth and reproduction of Pseudomonas, restraining the growth of microorganisms and preserve the quality of bigeye tuna. This study provides insights into understanding the effects of packaging material on maintaining quality during logistics transportation.

Crystal structures of Schistosoma mansoni histone deacetylase 8 reveal a novel binding site for allosteric inhibitors.

Parasitic diseases cause significant global morbidity and mortality particularly in the poorest regions of the world. Schistosomiasis, one of the most widespread neglected tropical diseases, affects more than 200 million people worldwide. Histone deacetylase (HDAC) inhibitors are prominent epigenetic drugs that are being investigated in the treatment of several diseases, including cancers and parasitic diseases. Schistosoma mansoni HDAC8 (SmHDAC8) is highly expressed in all life cycle stages of the parasite, and selective inhibition is required in order to avoid undesirable off-target effects in the host. Herein, by X-ray crystal structures of SmHDAC8-inhibitor complexes, biochemical and phenotypic studies, we found two schistosomicidal spiroindoline derivatives binding a novel site, next to Trp198, on the enzyme surface. We determined that by acting on this site, either by mutation of the Trp198 or by compound binding, a decrease in the activity of the enzyme is achieved. Remarkably, this allosteric site differs from the human counterpart; rather, it is conserved in all Schistosoma species, as well as Rhabidoptera and Trematoda classes, thus paving the way for the design of HDAC8-selective allosteric inhibitors with improved properties.

Gas-phase ion migration spectrum analysis of the volatile flavors of large yellow croaker oil after different storage periods.

The large yellow croaker, a species of fish found in the northwestern Pacific, is favored by consumers because of its prevalence in saltwater bodies, golden yellow abdomen, high calcium content, high protein, high fat content, and a flavor that originates from its lipids and volatile components. Volatile organic compounds significantly affect the aroma of food. In this work, electronic nose and headspace gas chromatography-ion mobility spectrometry were applied to analyze the flavor differences in fish oil durations. Through electronic nose system analysis, sensors W1C, W3S, W6S, and W2S directly affected fish oil flavor, and their flavor components were different. Gas chromatography-ion mobility spectrometry identified 26 volatile components (19 aldehydes, 3 ketones, 2 alcohols, 1 furan, and 1 olefin). (E,E)-2,4-hexadienal (D), (E,E)-2,4-hexadienal (M), 2,4-heptadienal (M), (E)-2-octenal, 2-propanone, 2-heptanone (M), 3-pentanone (D), and 1-octen-3-ol were the key flavor components of the fish oil. In conclusion, the combination of GC-IMS and PCA can identify the differences in flavor changes of large yellow croaker oil during 0-120 days storage. After 60 days storage, the types and signals of 2-propanone, 2-heptanone (M) components increase significantly. When 120 days storage, at this time, (E,E)-2,4-hexadienal (D), (E,E)-2,4-hexadienal (M), 2,4-heptadienal (M), (E)-2-octenal,(E)-2-octenal significantly. It has become the main flavor substance of fish oil. In summary, as the storage period increases, the components increase, and the oxidizing substances will increase, resulting in the deterioration of fish oil.

Comparative metabolic profiling by 1H-NMR spectroscopy analysis reveals the adaptation of S. mansoni from its host to in vitro culture conditions: a pilot study with ex vivo and GSH-supplemented medium-cultured parasites.

Schistosomiasis is a neglected tropical disease caused by parasitic flatworms (blood fluke) of the genus Schistosoma. Parasites acquire most nutrients for their development and sustainment within the definitive host either by ingestion into the gut or across the body surface. Over the years, the best conditions for long-term maintenance of parasites in vitro have been thoroughly established. In our hands, 1H-NMR spectroscopy represents a powerful tool to characterize the metabolic changes in S. mansoni in response to culturing condition perturbations. In order to compare the metabolic fingerprint of ex vivo and parasites cultured in vitro with or without the supplement of reduced glutathione, we conducted a pilot study applying the 1H-NMR spectroscopy-based metabolomics. We obtained new insight into specific metabolic pathways modulated under these different experimental conditions.

Edible Insects and Sustainable Development Goals.

The insect sector can become an important component of sustainable circular agriculture by closing nutrient and energy cycles, fostering food security, and minimising climate change and biodiversity loss, thereby contributing to SDGs. The high levels of the interaction of the insect sector with the SDGs is clearly illustrated inside the review, analysing all of the SDGs that can have direct and indirect effects on insects. Mapping the interactions between the SDGs goals and insect sector offers a starting point, from which it could be possible to define practical next steps for better insect policy.

Drug effects on metabolic profiles of Schistosoma mansoni adult male parasites detected by 1H-NMR spectroscopy.

Schistosomiasis is one of the most devastating neglected tropical parasitic diseases caused by trematodes of the genus Schistosoma. Praziquantel (PZQ) is today the only drug used in humans and animals for the treatment of schistosomiasis but unfortunately it is poorly effective on larval and juvenile stages of the parasite. Therefore, it is urgent the discovery of new drug targets and compounds. We have recently showed that the anti-anginal drug perhexiline maleate (PHX) is very active on multiple developmental stages of Schistosoma mansoni in vitro. It is well known that PHX impacts the lipid metabolism in mammals, but the final target on schistosomes still remains unknown. The aim of this study was to evaluate the ability of 1H nuclear magnetic resonance (NMR) spectroscopy in revealing metabolic perturbations due to PHX treatment of S. mansoni adult male worms. The effects of PHX were compared with the ones induced by vehicle and gambogic acid, in order to detect different metabolic profiles and specificity of the PHX action. Remarkably a list of metabolites associated to PHX-treatment was identified with enrichment in several connected metabolic pathways including also the Kennedy pathway mediating the glycerophospholipid metabolism. Our study represents the first 1H-NMR metabolomic approach to characterize the response of S. mansoni to drug treatment. The obtained "metabolic fingerprint" associated to PHX treatment could represent a strategy for displaying cellular metabolic changes for any given drug and to compare compounds targeting similar or distinct biochemical pathways.

Risks and critical issues related to the discovery on the market of unauthorized live alien species on the Italian territory: Chinese crab (Eriocheir sinensis).

Eriocheir sinensis, Chinese Crab or Chinese Mitten Crab is a catadromous species belonging to the Varunidae family, native to river and estuarine areas of North and South East China and Korea. At European level, E. sinensis is widespread in the main water basins of Central and Northern Europe and, since 2016, it has been included in the list of invasive species important for the European Union and subjected to confinement and eradication measures which include the prohibition of collection, transit and placing on the market of live specimens (Regulation (EC) N° 1143/2014). The Chinese Crab can represent a significant danger for the local ecosystem and for the native biota as well as contributing to the appearance of hydrogeological instability phenomena resulting from the intense excavation and erosion of the riverbanks. The first finding of 5 kg of live specimens of Eriocheir sinensis was recorded in the official control by the UFS (Functional Simple Unit) veterinary public health and food safety of the ASL Toscana Centro at an ethnic catering establishment. The specimens were subjected to seizure, photographed, identified morphologically, and subjected to euthanasia and destruction in accordance with the European requirements for welfare and management of animal by-products. From the sanitary point of view, the dangers associated with the consumption of this crab are mainly biological and chemical therefore, risk communication is fundamental, not only at the level of the competent authorities in the sector, but also for the food business operators.

Urban Consumer Trust and Food Certifications in China.

China has experienced frequent food safety incidents that have undermined consumer trust in the food supply chain. To overcome this problem, China requalified the legislative framework and adopted a comprehensive food certification system over the years. Here, we investigated the influences of food traceability and Chinese certifications (QS/SC-food quality safety market access/production system, hazard-free, green, and organic) on Chinese consumer trust of food safety for different types of products: fish, meat, milk, eggs, and rice. Data were collected through face-to-face surveys conducted in rural and urban Chinese areas. With a sample of 757 questionnaires, we ran a logit model. The results show consumers' uncertainty and skepticism of certifications guaranteeing food safety attributes, especially for animal-based products. We found that price is used as a cue of safety by Chinese consumers. Individuals with higher education seem less influenced by certifications and other cues included in the analysis. The findings demonstrate that Chinese policy makers should implement new strategies to enhance consumer food safety trust, and design policies by considering different categories (e.g., vegetables, meat, fish, etc.) of food.

(+)-(R)- and (-)-(S)-Perhexiline maleate: Enantioselective synthesis and functional studies on Schistosoma mansoni larval and adult stages.

Schistosomiasis is a neglected tropical disease mainly affecting the poorest tropical and subtropical areas of the world with the impressive number of roughly 200 million infections per year. Schistosomes are blood trematode flukes of the genus Schistosoma causing symptoms in humans and animals. Organ morbidity is caused by the accumulation of parasite eggs and subsequent development of fibrosis. If left untreated, schistosomiasis can result in substantial morbidity and even mortality. Praziquantel (PZQ) is the most effective and widely used compound for the treatment of the disease, in prevention and control programs in the last 30 years. Unfortunately, it has no effect on juvenile immature schistosomes and cannot prevent reinfection or interfere with the schistosome life cycle; moreover drug-resistance represents a serious threat. The search for an alternative or complementary treatment is urgent and drug repurposing could accelerate a solution. The anti-anginal drug perhexiline maleate (PHX) has been previously shown to be effective on larval, juvenile, and adult stages of S. mansoni and to impact egg production in vitro. Since PHX is a racemic mixture of R-(+)- and S-(-)-enantiomers, we designed and realized a stereoselective synthesis of both PHX enantiomers and developed an analytical procedure for the direct quantification of the enantiomeric excess also suitable for semipreparative separation of PHX enantiomers. We next investigated the impact of each enantiomer on viability of newly transformed schistosomula (NTS) and worm pairs of S. mansoni as well as on egg production and vitellarium morphology by in vitro studies. Our results indicate that the R-(+)-PHX is mainly driving the anti-schistosomal activity but that also the S-(-)-PHX possesses a significant activity towards S. mansoni in vitro.

Luminescence-Based, Low- and Medium-Throughput Assays for Drug Screening in Schistosoma mansoni Larval Stage.

Schistosomiasis is one of the major parasitic diseases with more than  200 million people infected worldwide every year. Praziquantel is the drug of choice against the schistosomiasis although the use of a single drug to treat such a large amount of infected people appears particularly worrisome. For this reason, the search of new schistosomicidal compounds is viewed as an urgent goal and a number of screening campaigns have been carried out in the past years. The larval stage of Schistosoma (schistosomula) has been widely used in order to identify new compounds against the parasite. Here we describe detailed practical procedures for a luminescence-based assay proven to be highly effective for the selection of schistosomicidal compounds on small and medium-high scale. The assay is based on the quantitation of the parasite ATP, a good indicator of metabolically active cells, as measure of schistosomula viability. This assay is fast and reproducible, and it is suitable either for manual or for semiautomated screenings.

Investigating the Antiparasitic Potential of the Marine Sesquiterpene Avarone, Its Reduced form Avarol, and the Novel Semisynthetic Thiazinoquinone Analogue Thiazoavarone.

The chemical analysis of the sponge Dysidea avara afforded the known sesquiterpene quinone avarone, along with its reduced form avarol. To further explore the role of the thiazinoquinone scaffold as an antiplasmodial, antileishmanial and antischistosomal agent, we converted the quinone avarone into the thiazinoquinone derivative thiazoavarone. The semisynthetic compound, as well as the natural metabolites avarone and avarol, were pharmacologically investigated in order to assess their antiparasitic properties against sexual and asexual stages of Plasmodium falciparum, larval and adult developmental stages of Schistosoma mansoni (eggs included), and also against promastigotes and amastigotes of Leishmania infantum and Leishmania tropica. Furthermore, in depth computational studies including density functional theory (DFT) calculations were performed. A toxic semiquinone radical species which can be produced starting both from quinone- and hydroquinone-based compounds could mediate the anti-parasitic effects of the tested compounds.

Screening and Phenotypical Characterization of Schistosoma mansoni Histone Deacetylase 8 (SmHDAC8) Inhibitors as Multistage Antischistosomal Agents.

Schistosomiasis (also known as bilharzia) is a neglected tropical disease caused by platyhelminths of the genus Schistosoma. The disease is endemic in tropical and subtropical areas of the world where water is infested by the intermediate parasite host, the snail. More than 800 million people live in endemic areas and more than 200 million are infected and require treatment. Praziquantel (PZQ) is the drug of choice for schistosomiasis treatment and transmission control being safe and very effective against adult worms of all the clinically relevant Schistosoma species. Unfortunately, it is ineffective on immature, juvenile worms; therefore, it does not prevent reinfection. Moreover, the risk of development and spread of drug resistance because of the widespread use of a single drug in such a large population represents a serious threat. Therefore, research aimed at identifying novel drugs to be used alone or in combination with PZQ are needed. Schistosoma mansoni histone deacetylase 8 (SmHDAC8) is a class I zinc-dependent HDAC, which is abundantly expressed in all stages of its life cycle, thus representing an interesting target for drug discovery. Through virtual screening and phenotypical characterization of selected hits, we discovered two main chemical classes of compounds characterized by the presence of a hydroxamate-based metal binding group coupled to a spiroindoline or a tricyclic thieno[3,2-b]indole core as capping groups. Some of the compounds of both classes were deeply investigated and showed to impair viability of larval, juvenile, and adult schistosomes, to impact egg production in vitro and/or to induce morphological alterations of the adult schistosome reproductive systems. Noteworthy, all of them inhibit the recombinant form of SmHDAC8 enzyme in vitro. Overall, we identified very interesting scaffolds, paving the way to the development of effective antischistosomal agents.

Thiazinoquinones as New Promising Multistage Schistosomicidal Compounds Impacting Schistosoma mansoni and Egg Viability.

Schistosomiasis is the most significant neglected tropical parasitic disease caused by helminths in terms of morbidity and mortality caused by helminths. In this work, we present the antischistosomal activity against Schistosoma mansoni of a rationally selected small set of thiazinoquinone derivatives, some of which were previously found to be active against Plasmodium falciparum and others synthesized ad hoc. The effects on larvae, juvenile, and adult parasite viability as well as on egg production and development were investigated, resulting in the identification of new multistage antischistosomal hit compounds. The most promising compounds 6, 8, 13, and 14 with a LC50 value on schistosomula from ∼5 to ∼15 µM also induced complete death of juvenile (28 days old) and adult worm pairs (7 weeks old) and a detrimental effect on egg production and development in vitro. Structure-activity relationships (SARs) were analyzed by means of computational studies leading to the hypothesis of a redox-based mechanism of action with a one-electron reduction bioactivation step and the subsequent formation of a toxic semiquinone species, similarly to what was previously observed for the antiplasmodial activity. Our results also evidenced that the selective toxicity against mammalian cells or parasites as well as specific developmental stages of a parasite can be addressed by varying the nature of the introduced substituents.

Identification of novel multi-stage histone deacetylase (HDAC) inhibitors that impair Schistosoma mansoni viability and egg production.

BACKGROUND: Novel anti-schistosomal multi-stage drugs are needed because only a single drug, praziquantel, is available for the treatment of schistosomiasis and is poorly effective on larval and juvenile stages of the parasite. Schistosomes have a complex life-cycle and multiple developmental stages in the intermediate and definitive hosts. Acetylation and deacetylation of histones play pivotal roles in chromatin structure and in the regulation of transcription in eukaryotic cells. Histone deacetylase (HDAC) inhibitors modulate acetylation of several other proteins localized both in the nucleus and in the cytoplasm and therefore impact on many signaling networks and biological processes. Histone post-translational modifications may provide parasites with the ability to readily adapt to changes in gene expression required for their development and adaptation to the host environment. The aim of the present study was to screen a HDAC class I inhibitor library in order to identify and characterize novel multi-stage hit compounds. METHODS: We used a high-throughput assay based on the quantitation of ATP in the Schistosoma mansoni larval stage (schistosomula) and screened a library of 1500 class I HDAC inhibitors. Subsequently, a few hits were selected and further characterized by viability assays and phenotypic analyses on adult parasites by carmine red and confocal microscopy. RESULTS: Three compounds (SmI-124, SmI-148 and SmI-558) that had an effect on the viability of both the schistosomula larval stage and the adult worm were identified. Treatment with sub-lethal doses of SmI-148 and SmI-558 also decreased egg production. Moreover, treatment of adult parasites with SmI-148, and to a lesser extent Sm-124, was associated with histone hyperacetylation. Finally, SmI-148 and SmI-558 treatments of worm pairs caused a phenotype characterized by defects in the parasite reproductive system, with peculiar features in the ovary. In addition, SmI-558 induced oocyte- and vitelline cell-engulfment and signs of degeneration in the uterus and/or oviduct. CONCLUSIONS: We report the screening of a small HDAC inhibitor library and the identification of three novel compounds which impair viability of the S. mansoni larval stage and adult pairs. These compounds are useful tools for studying deacetylase activity during parasite development and for interfering with egg production. Characterization of their specificity for selected S. mansoni versus human HDAC could provide insights that can be used in optimization and compound design.

DNA barcoding for the verification of supplier's compliance in the seafood chain: How the lab can support companies in ensuring traceability.

Food Business Operators (FBOs) rely on laboratory analysis to ensure seafood traceability. DNA barcoding and Forensically Informative Nucleotide Sequencing may represent a support within self-checking programs finalized to suppliers' qualification and products identity certification. The present study aimed at verifying the usefulness of a decisional procedure (decision tree) set up at the FishLab (Department of Veterinary Sciences, University of Pisa, Italy) for seafood species identification by DNA analysis, to cope with FBOs' needs. The decision tree was applied to the analysis of 182 seafood (fish and molluscs) products, conferred to the FishLab by different FBOs between 2014 and 2015 as result of their self-checking activities. The analysis relied on a standard COI gene fragment eventually integrated by the analysis of alternative or supportive molecular targets (cytb and 16S rRNA). It also included a mini-DNA barcoding approach for processed products. Overall, 96.2% of the samples were unambiguously identified at species level using the elective target alone (92.4%) or a multi-target approach (3.8%). The lack of species identification (3.8%) was attributable to the absence of reference sequences or to the low resolution of the molecular targets. Nonetheless, all the molecular results were deemed adequate to evaluate the sample's compliance to the label information. Non-compliances were highlighted in 18.1% of the products. The protocol was proven as an effective supportive tool for the seafood identity verification within the supply chain self-checking activities. In addition, a considerable fraud rate was confirmed and the species most frequently involved in substitution were pointed out.

Discovery by organism based high-throughput screening of new multi-stage compounds affecting Schistosoma mansoni viability, egg formation and production.

Schistosomiasis, one of the most prevalent neglected parasitic diseases affecting humans and animals, is caused by the Platyhelminthes of the genus Schistosoma. Schistosomes are the only trematodes to have evolved sexual dimorphism and the constant pairing with a male is essential for the sexual maturation of the female. Pairing is required for the full development of the two major female organs, ovary and vitellarium that are involved in the production of different cell types such as oocytes and vitellocytes, which represent the core elements of the whole egg machinery. Sexually mature females can produce a large number of eggs each day. Due to the importance of egg production for both life cycle and pathogenesis, there is significant interest in the search for new strategies and compounds not only affecting parasite viability but also egg production. Here we use a recently developed high-throughput organism-based approach, based on ATP quantitation in the schistosomula larval stage of Schistosoma mansoni for the screening of a large compound library, and describe a pharmacophore-based drug selection approach and phenotypic analyses to identify novel multi-stage schistosomicidal compounds. Interestingly, worm pairs treated with seven of the eight compounds identified show a phenotype characterized by defects in eggshell assemblage within the ootype and egg formation with degenerated oocytes and vitelline cells engulfment in the uterus and/or oviduct. We describe promising new molecules that not only impair the schistosomula larval stage but also impact juvenile and adult worm viability and egg formation and production in vitro.

Detection of selected trace elements in yogurt components.

The objective of this study was to determine the concentrations of Cu, Cd, Pb, Mn, Cr, Co, Ni, Zn, and Hg in the white and fruit parts of commercially available yogurts (n = 30) from Nitra markets (Slovak Republic). The results were correlated to determine their relationships. Three yogurt fruit flavors were chosen and tested, strawberry (n = 10), blueberry (n = 10), and cherry (n = 10). The elements were analyzed using atomic absorption spectrophotometry. Higher concentrations of toxic elements, such as Cd and Pb, were found in the fruit parts of the yogurt, and in some cases, the tolerable limit was exceeded. The white part of the yogurt was not contaminated by toxic elements. White yogurt is a good source of nutrients for humans, but the fruit part in yogurt requires detailed monitoring and improvements in the processing techniques.

Meat from cattle slaughtered without stunning sold in the conventional market without appropriate labelling: A case study in Italy.

In the European Union, slaughter without stunning is allowed for religious slaughter to obtain halal and kosher meat. Especially in the case of Jewish slaughtering, cuts which are not deemed as kosher are sold to regular market without any specific labelling. This survey, conducted in Tuscany in 2016, aimed to quantify the carcasses rejected in relation to the type of religious slaughter. 656 bovines were slaughtered without stunning: 538 (82%) for halal and 118 (18%) for kosher. All carcasses slaughtered by the Islamic procedure (dhabiha) were considered halal, while 77.1% of carcasses slaughtered by the Jewish procedure (shechita) did not pass the approval. Carcasses were rejected after chest cavity inspection (50%) and after the lungs control (50%). This study provides an important insight in this field and postulates how to amalgamate the concepts of freedom of religion, as enshrined by the Charter of Fundamental Rights of the EU, with consumer rights and animal welfare.