Short communication: Detection of undeclared presence of bovine milk in buffalo yogurt.

The authenticity of buffalo (Bubalus bubalis) dairy products is a focal issue, considering the increasing demand for buffalo milk products. Therefore, the aim of this study was to investigate the undeclared presence of bovine (Bos taurus) milk in buffalo yogurt, to understand which risk factors might make the product vulnerable to fraud. Real-time PCR assay showed the undeclared presence of bovine DNA in addition to buffalo DNA in 18 of 72 samples. Given the widespread lack of data on the presence of undeclared milk species in buffalo dairy products, the study provides a significant insight into the incidence of fraud in the buffalo dairy field. The data from this study could help improve the analysis of food safety risks along the buffalo milk supply chain and in the dairy processing industry, perceived as being highly vulnerable to food fraud, and prioritize target areas for food policy making to steer and enforce European food fraud regulations.

Occurrence of potentially pathogenic arcobacters in shellfish.

Considering that several recent cases of human gastroenteritis have been associated with species from the Arcobacter genus, and that few data are currently available about the occurrence of this genus in Italian shellfish, the aim of the present study was to evaluate the occurrence of Arcobacter spp. and the presence of virulence-associated genes. The approach consisted of cultural and biomolecular (multiplex-PCR and 16S-RFLP) methods identifying isolates, followed by PCR assays aimed at the cadF, ciaB, cjl349, irgA, hecA putative virulence genes. Arcobacter spp. was detected in 16/70 (22.8%) shellfish samples. Specifically, Arcobacter spp. was highlighted in 10/42 (23.8%) mussel and in 6/28 (21.4%) clam samples. Subsequently, biomolecular assays revealed Arcobacter butzleri in 12/16 (75%) and Arcobacter cryaerophilus 1B in 4/16 (25%) isolates. PCRs aimed at the five putative virulence genes demonstrated widespread distribution of these genes among Arcobacter isolates and some differences from the results published by other authors. Our research provides more information regarding the health risks associated with the consumption of raw bivalve molluscs and underlines the need to implement an adequate control plan by performing intensive and continuous monitoring in order to guarantee human health.

Occurrence of mislabeling in meat products using DNA-based assay.

Considering that the authentication of food contents is one of the most important issues for the food quality sector, and given the increasing demand for transparency in the meat industry followed the horsemeat scandal in Europe, this study investigates processed-meat products from Italian markets and supermarkets using the mitochondrial cytochrome b gene qualitative PCR identification system in order to verify any species substitution or mislabeling. The results revealed a high substitution rate among the meat products, highlighting a mislabeling rate of 57 %, and consequently, considerable discordance with the indications on the labels, which raises significant food-safety and consumer-protection concerns.

Occurrence and characterization of Arcobacter spp. from ready-to-eat vegetables produced in Southern Italy.

Given that the number of foodborne illness outbreaks linked to the consumption of ready-to-eat vegetables has been widely documented and considering that data on the occurrence of Arcobacter spp. in such foodstuffs are lacking, the aim of the present study was to evaluate the presence of Arcobacter spp. and the occurrence of virulence factors as well as to genotype Arcobacter spp. in ready-to-eat (RTE) vegetable samples, using cultural and biomolecular assays. Arcobacter spp. was detected in 16/110 (14.5%) samples, with A. butzleri being detected in 15/16 and A. cryaerophilus in 1/16 isolates. PCRs aimed at the nine putative virulence genes demonstrated widespread distribution of such genes among A. butzleri and A. cryaerophilus isolates. In addition, multilocus sequence type (MLST) analysis revealed a low genetic diversity within the arcobacters isolates. The results underline the need to develop an appropriate surveillance system based on biomolecular characterization for an integrated microbiological risk assessment of ready-toeat vegetables, and consequently of composite foods.

Determining the Authenticity of Shark Meat Products by DNA Sequencing.

Given that the global shark meat market is poised to grow in future years, the aim of this study was to use DNA sequencing of the cytochrome c oxidase I (COI) and NADH dehydrogenase subunit 2 (NADH2) mitochondrial genes to examine the market of shark meat products in Italy. This made it possible to analyze patterns of species utilization and commercialization of threatened, endangered and/or prohibited species, focusing on fraudulent activities in the shark food chain in order to propose seafood safety and environmental sustainability solutions. The study shows that the labeling of shark meat products generally lacks comprehensive information, thus making it difficult for consumers to make informed purchasing decisions and fails to comply with European Union (EU) legislation regarding seafood labelling. Molecular investigation reveals a high mislabeling rate (45.4%), highlighting widespread use of cheaper species either in order to replace species that are better known and more popular, or else in order to sell various threatened species. Considering that seafood mislabeling can circumvent the management of sustainable fisheries and facilitate Illegal, Unreported and Unregulated (IUU) fishing, the routine use of genetic analysis should be encouraged among control and enforcement agencies in order to implement effective management measures. This would help to build a species-specific reporting system for all catches, and enhance control measures, in order to prevent illegal activities connected with shark catches and trade around the world.

Evaluation of the animal welfare during religious slaughtering.

Plasma cortisol and its metabolites are physiological indicators for stress assessment and slaughtering method may affect their levels, playing an important role in the correct acidification of meat. The aim of the study was to determine and compare plasma cortisol values in animals slaughtered using traditional procedures, which include stunning (using captive bolt pistol), with those in animals slaughtered using Halal method, which does not involve stunning. The study was carried out on a total of 60 Charolais male beef cattle of eight months of age, bred in free paddock outdoors. The animals were divided into two experimental groups, each consisting of 30 individuals, on the basis of the slaughtering method, i.e. traditional or Halal, to verify the whole production chain and to ensure that the product conformed to Muslim rules. Plasma cortisol levels (detected by Elisa test) were evaluated at two different times of animal productive life: on the farm, one week before slaughter (T0) and during bleeding (T1). The 30 calves slaughtered after stunning showed plasma cortisol values of 4.06±1.94 and 43.72±12.09 nmol/L, respectively on the farm and during exsanguination. Conversely, the average values found in the 30 calves subjected to ritual slaughter were 3.26±1.01 and 88.81±41.02 nmol/L. The study demonstrated that throughout the animal's productive life (from pasture to slaughter) the greatest variation between slaughter with and without stunning was observed during bleeding.

Author Correction: Sequence-defined multifunctional polyethers via liquid-phase synthesis with molecular sieving.

In the version of this Article originally published, the authors inadvertently cited ref. 10 in two places in the first paragraph. They would like to clarify that it should not have been cited in the sentence that starts "Polymer chemists have employed strategies such as single monomer insertion..." as it mistakenly implied that the IEG+ method described in ref. 10 could not produce unimolecular polymers; it can do so, as was demonstrated in ref. 10. The authors would also like to clarify that ref. 10 should not have been cited in the sentence that starts "Moreover, solid-phase synthesis is generally difficult to scale up...", as it implied that ref. 10 uses solid-phase synthesis; it does not, and is a purely liquid-phase process. The citation of ref. 10 has now been removed from these two sentences, but has been included elsewhere in the first two paragraphs of the Article as follows. In the first paragraph, at the end of the sentence "In iterative synthesis, specific monomers are added one at a time, or as multiples, to the end of a growing polymer chain, then reaction debris is separated from the chain extended polymer, and the cycle is repeated using the next monomer in the sequence10-12."; this sentence has been further amended to indicate multiple monomers can also be added. The reference has also been added to the end of the first sentence of the second paragraph, which starts "Consequently, liquid-phase iterative synthetic methods...", and in the third sentence of that paragraph, which now starts "For example, Johnson10, Whiting....".

Sequence-defined multifunctional polyethers via liquid-phase synthesis with molecular sieving.

Synthetic chemists have devoted tremendous effort towards the production of precision synthetic polymers with defined sequences and specific functions. However, the creation of a general technology that enables precise control over monomer sequence, with efficient isolation of the target polymers, is highly challenging. Here, we report a robust strategy for the production of sequence-defined synthetic polymers through a combination of liquid-phase synthesis and selective molecular sieving. The polymer is assembled in solution with real-time monitoring to ensure couplings proceed to completion, on a three-armed star-shaped macromolecule to maximize efficiency during the molecular sieving process. This approach is applied to the construction of sequence-defined polyethers, with side-arms at precisely defined locations that can undergo site-selective modification after polymerization. Using this versatile strategy, we have introduced structural and functional diversity into sequence-defined polyethers, unlocking their potential for real-life applications in nanotechnology, healthcare and information storage.

Analysis of Stress Indicators for Evaluation of Animal Welfare and Meat Quality in Traditional and Jewish Slaughtering.

Sixty Charolais male beef cattle of eight months of age were divided into two groups according to the slaughtering method, i.e., traditional or Kosher (religious Jewish rite). The aim of the study was to detect and compare the plasma concentrations of cortisol and catecholamines (dopamine, norepinephrine and epinephrine), by Elisa and HPLC test. These four stress indicators were evaluated during three different stages of each animal productive life: on the farm (step 1), after transportation (step 2) and during bleeding (step 3). The patterns of the parameters measured were similar and, interestingly, revealed significant changes throughout the three steps considered. The greatest variation between the two methods of slaughtering was observed in step 3, where we found a statistically significant difference with all the parameters except epinephrine. In the animals slaughtered by the religious rite, cortisol, dopamine, norepinephrine and epinephrine were 68.70 ± 30.61 nmol/L; 868.43 ± 508.52 ng/L; 3776.20 ± 1918.44 ng/L; and 4352.20 ± 3730.15 ng/L, respectively, versus 45.08 ± 14.15 nmol/L; 513.87 ± 286.32 ng/L; 3425.57 ± 1777.39 ng/L; and 3279.97 ± 1954.53 ng/L, respectively, in the other animals. This suggests that the animals slaughtered by the Kosher rite are subjected to higher stress conditions at the exsanguination phase. The animals slaughtered by the religious Jewish rite showed lower cortisol and catecholamine levels on the farm (step 1) and after transportation to the slaughterhouse (step 2). This was likely because the animals selected at the end of step 1 by the Rabbis for the religious rite are usually the most docile and gentle.

The Selectivity Challenge in Organic Solvent Nanofiltration: Membrane and Process Solutions.

Recent development of organic solvent nanofiltration (OSN) materials has been overwhelmingly directed toward tight membranes with ultrahigh permeance. However, emerging research into OSN applications is suggesting that improved separation selectivity is at least as important as further increases in membrane permeance. Membrane solutions are being proposed to improve selectivity, mostly by exploiting solute/solvent/membrane interactions and by fabricating tailored membranes. Because achieving a perfect separation with a single membrane stage is difficult, process engineering solutions, such as membrane cascades, are also being advocated. Here we review these approaches to the selectivity challenge, and to clarify our analysis, we propose a selectivity figure of merit that is based on the permselectivity between the two solutes undergoing separation as well as the ratio of their molecular weights.

Determination of plasmatic cortisol for evaluation of animal welfare during slaughter.

The plasmatic cortisol levels of 60 eight-month-old calves (Charolais breed) were measured as stress indicators resulting from two types of slaughter: traditional and religious rite. The plasmatic cortisol levels were evaluated during three different stages of their productive life: during growth, after transport and during slaughter. The thirty calves slaughtered after stunning showed plasmatic cortisol values of 4.85±3.2; 36.36±12.2 and 45.08±14.1 nmol/L, during growth, in the slaughterhouse stables and during exsanguination, respectively. Conversely, the average values found in the thirty calves subjected to ritual slaughter were 2.96±1.2; 31.65±25.4 and 68.70±30.6 nmol/L. The results of the study showed that animal welfare should be improved in both forms of slaughter.

DNA-based approach for species identification of goat-milk products.

Considering that mislabeled milk products have been widely reported throughout the world and that the authentication of food components is one of the key issues in food safety and quality, the aim of this study was to use DNA-based methods to investigate the prevalence of mislabeling among goat-milk products and, consequently, how far the ingredients matched the labels. The study reveals a high degree of species mislabeling in milk products (80%), underlining the need to enhance dairy traceability practices, so as to guarantee product authenticity, and provide reliable information to consumers.

Occurrence of emerging food-borne pathogenic Arcobacter spp. isolated from pre-cut (ready-to-eat) vegetables.

Given that changes in consumer food behaviours have led to an increase in the demand for pre-cut ready-to-eat (RTE) vegetables, and that few data are currently available on the occurrence of Arcobacter spp. in such foods, the aim of the present study was to assess the occurrence of Arcobacter spp. that carry virulence-associated genes on pre-cut RTE vegetables, using cultural and molecular methods. Arcobacter was detected using biomolecular identification methods in 44/160 (27.5%) of the samples, of which 40/44 (90.9%) isolates corresponded to A. butzleri and 4/44 (9.1%) to A. cryaerophilus. Studying the incidence of 9 virulence-associated genes revealed the widespread distribution of these genes among the Arcobacter isolates tested. The results obtained in our research provided plenty of information on the health risks associated with the direct consumption of raw vegetables, and highlight the need to implement further studies at each level of the production chain, in order to obtain further information to help protect human health.

Packaged frozen fishery products: species identification, mislabeling occurrence and legislative implications.

Given the increase in the international trade of packaged frozen fishery products, this study used DNA barcoding to investigate the breaded hake and plaice species, sold in Italian markets. The results of this study generally matched the ingredient list on the food label. Only 6 of the 120 samples were non-compliant. Specifically, breaded merluccius samples match the species shown in the list of ingredients on the label. Of the "breaded plaice" samples, 4/14 contained Lepidopsetta polyxystra and 2/14 Merluccius gayi, thus failing to match the ingredient list on the label. Considering the European legislation indicates that the label must not mislead consumers, but international trade and the use of similar terms for different products makes it complicated when a product from one country is introduced into another in which the niche already exists, clear labeling is strongly recommended in order to ensure that consumers can make conscious choices.

Tunable-Porosity Membranes From Discrete Nanoparticles.

Thin film composite membranes were prepared through a facile single-step wire-wound rod coating procedure in which internally crosslinked poly(styrene-co-butadiene) polymer nanoparticles self-assembled to form a thin film on a hydrophilic ultrafiltration support. This nanoparticle film provided a defect-free separation layer 130-150 nm thick, which was highly permeable and able to withstand aggressive pH conditions beyond the range of available commercial membranes. The nanoparticles were found to coalesce to form a rubbery film when heated above their glass transition temperature (Tg). The retention properties of the novel membrane were strongly affected by charge repulsion, due to the negative charge of the hydroxyl functionalized nanoparticles. Porosity was tuned by annealing the membranes at different temperatures, below and above the nanoparticle Tg. This enabled fabrication of membranes with varying performance. Nanofiltration properties were achieved with a molecular weight cut-off below 500 g mol(-1) and a low fouling tendency. Interestingly, after annealing above Tg, memory of the interstitial spaces between the nanoparticles persisted. This memory led to significant water permeance, in marked contrast to the almost impermeable films cast from a solution of the same polymer.

Occurrence of Mislabelling in Prepared Fishery Products in Southern Italy.

Fish authentication is a major concern not only for the prevention of commercial fraud, but also for the assessment of safety risks deriving from the undeclared introduction of potentially dangerous toxic or allergenic substances or environmentally damaging fish where endangered species are involved. Moreover, food authentication might affect the diet of certain groups of consumers, such as followers of religious practices. Considering the authentication of fish products is one of the key issues in food safety, quality and sustainability, the aim of this work was to investigate the prevalence of mislabelling in sole (Solea solea), plaice (Pleuronectes platessa), Atlantic salmon (Salmo salar), and hake (Merluccius merluccius) fillets from markets and supermarkets located in Apulia (Southern Italy) using DNA barcoding. The results of the molecular investigations reveal that 42/98 (42.8%) fillet samples were not correctly labelled. In particular, 12/27 (44.4%) fillets of sole (Solea solea) were identified as belonging to Solea senegalensis. In addition, 13/28 (46.4%) plaice (Pleuronectes platessa) samples were identified as Pangasius hypophtalmus. All Atlantic salmon (Salmo salar) samples were correctly labelled. Post-sequencing data analysis revealed that 17/30 (56.6%) hake fillets (Merluccius merluccius) were not correctly labelled, of which 8/30 samples identified as Merluccius hubbsi, 5/30 samples as Merluccius products and 4/30 as Merluccius capensis. The study reveals a high occurrence of species mislabelling in the prepared fish fillet products, further evidence of the need for increased traceability and assessment of the authenticity of food products.