Application of RNA-sequencing to identify biomarkers in broiler chickens prophylactic administered with antimicrobial agents.

Antimicrobial (AM) resistance is largely acknowledged as one of the biggest global health and food safety challenges and the overuse of AMs is known to generate resistance in bacteria that may affect both animals and humans. Poultry meat is the second most-produced meat in the European Union and in recent years consumers are becoming more concerned about food safety, traceability, and animal welfare in poultry rearing system, increasingly requiring meats from broilers reared without AMs. In the present study, we performed RNA sequencing to analyze 64 liver and 54 muscle transcriptomic profiles in broilers reared without treatment or treated with different classes of AMs. Moreover, we validated the most differentially expressed genes among the treated groups to detect putative novel biomarkers able to discriminate meats of broilers reared without AMs. The PDK4, IGFBP1, and RHOB genes were identified as putative novel hepatic biomarkers, discriminating broilers treated with AMs compared to broilers reared without treatments. The whole transcriptome changes revealed the liver as a valuable target organ for AM administration screening. In addition, our results suggest a leading effect of the coccidiostat when associated with AMs, influencing several biological processes. Our study showed that RNA sequencing is a powerful and valuable method to detect aberrant regulated genes and to identify biomarker candidates for AM misuse detection in farm animals. Further validation on larger sample size and a wider spectrum of AMs are needed to confirm the viability of the aforementioned biomarkers in poultry population.

Feasibility of biodegradable based packaging used for red meat storage during shelf-life: A pilot study.

This study was designated to ascertain the effectiveness of polylactic acid (PLA) based packaging solution to store red fresh meat during its refrigerated shelf-life. Recently the attention in the packaging industry regarding the use of bioplastics has been shifting from compostable/biodegradable materials toward biobased materials. Steaks obtained from semimembranous muscle of Piemontese beef were packaged in PLA trays closed with a lid made of PLA film and for comparison purposed in a conventional reference package consisting of a amorphous polyethylene terephthalate/polyethylene (APET/PET) trays and wrapped in plastic film of polyvinyl chloride (PVC). The packaging under modified atmosphere MAP was carried out by using a gas mixture of 66% O2, 25% CO2 and 9%N2. By using PLA packaging combination it was possible to maintain an optimum red colour together with a reduced content of volatile compounds associated to off-flavours of meat samples particularly related to the oxidation phenomena.

Mussels and clams from the italian fish market. is there a human exposition risk to metals and arsenic?

Seafood is associated with many beneficial effects on human health. However, the overall level of contaminants in biota has increased over the last two centuries and seafood is one of the source of oral exposition to contaminants. Therefore, this work aimed to evaluate cadmium, lead, mercury, arsenic, chromium and nickel presence in mussels and clams, from the Italian market, and the associated risk. The samples were from five different FAO areas. Analyses were carried out using inductively-coupled plasms-mass spectrometry. The sample concentrations were below the maximum levels stated by Commission Regulation (EC) 1881/2006, except one mussel sample, which was non-compliant for cadmium (2.13 ± 0.20 mg kg-1). For arsenic, nickel and chromium, maximum levels are not stated by the European Union. In this study, arsenic ranged from 1.29 to 13.35 mg kg-1 and nickel ranged from 1, and BMDL10 for lung bladder and skin cancer in all mussel samples was overcome, in the 100% and 25% of mussel and clam samples, respectively.

The occurrence of pesticides and persistent organic pollutants in Italian organic honeys from different productive areas in relation to potential environmental pollution.

Bee products, such as honey, are widely consumed as food and consumer interest is currently oriented towards organic foods. Regarding this, the European Commission establishes that the qualification of organic honey and other beekeeping products as being from organic production is closely bound with the characteristics of hive treatments as well as the quality of the environment. Agricultural contamination with pesticides is a challenging problem that needs to be fully addressed, in particular in the field of organic production systems. In this study, the occurrence of different classes of contaminants selected as representative of potential contamination sources were investigated in 59 organic honeys: organochlorines, OCs; organophosphates, OPs; polychlorobiphenyls, PCBs and polybromodiphenylethers, PBDEs. A method based on Accelerated Solvent Extraction with "in line" clean-up and GC-MS/MS detection was developed to detect contaminants. Residues of many pesticides were found in most of the samples investigated. The majority of honey samples contained at least one of the pesticides, even if their concentrations were found to be lower than its MRL. Diazinon, Mevinphos, Coumaphos, Chlorpyrifos and Quinoxyfen were the residues frequently detected in samples coming from the apple and citrus orchard areas. Furthermore, the results of the present study show that the presence of the residue in organic honey may also be affected by the geographical area (e.g. the presence of an agricultural system) confirming honey bee and beehive matrices as appropriate sentinels for monitoring contamination in the environment. The optimised method proved to be simple and rapid, requiring small sample sizes and minimising solvent consumption, due to the ASE having an "in line" clean-up step.

Distribution of persistent organic pollutants (POPS) IN wild Bluefin tuna (Thunnus thynnus) from different FAO capture zones.

Residues of environmental contaminants in food represent a concern in food safety programs. In this study, the distribution of persistent organic pollutants (POPs) were evaluated in 79 tuna samples from FAO areas 51 (Indian Ocean), 71 (Pacific Ocean), 34 (Atlantic Ocean), and 37 (Mediterranean Sea). 6 polychlorinated biphenyls (PCBs), 16 organochlorines (OCs) and 7 polybrominated biphenyl ethers (PBDEs) were selected as representative compounds according to EFSA POPs monitoring guidelines. An analytical method, based on Accelerated Solvent Extraction (ASE), with an "in-line" clean-up step and GC-MS/MS detection, was developed, validated and applied. PCBs were detected in all FAO areas, with a prevalence of 100% for most of them. In the FAO area 37, only, all PBDEs were detected. Only 5 OCs were detected. The results showed that POPs contamination of tuna reflects FAO area contamination; in particular FAO area 37 was the most polluted. Moreover, tuna muscle was an appropriate matrix for monitoring contamination and for obtaining information about food safety.

Effect of commercial or depurinized milk on rat liver growth-regulatory kinases, nuclear factor-kappa B, and endonuclease in experimental hyperuricemia: comparison with allopurinol therapy.

Hyperuricemia is a biochemical hallmark of gout, renal urate lithiasis, and inherited purine disorders, and may be a result of enormous ATP breakdown or purine release as a result of cardiovascular disease, hypertension, kidney disease, eclampsia, obesity, metabolic syndrome, psoriasis, tumor lysis syndrome, or intense physical training. The beneficial role of dairy products on hyperuricemia management and prevention is well documented in the literature. The primary aim of our experimental study was to examine the effect of milk dietary regimen (commercial 1.5% fat UHT milk or patented depurinized milk) compared with allopurinol therapy on experimental hyperuricemia induced by oxonic acid in rats. Principal component analysis was applied on a data set consisting of 11 variables for 8 different experimental groups. Among the 11 parameters measured (plasma uric acid and the liver parameters NFκB-p65, Akt kinase/phospho-Akt kinase, ERK kinase/phospho-ERK kinase, IRAK kinase/phospho IRAK kinase, p38/phospho-p38, and DNase), Akt/phospho Akt and ERK/phospho-ERK signaling were extracted as the most discriminating. We also compared the content of various potentially toxic compounds (sulfur compounds, ketones, aldehydes, alcohols, esters, carboxylic acids, and phthalates) in untreated commercial milk and depurinized milk. Of all the compounds investigated in this study that were observed in commercial milk (24 volatile organic compounds and 4 phthalates), 6 volatile organic compounds were not detected in depurinized milk. For almost all of the other compounds, significant decreases in concentration were observed in depurinized milk compared with commercial milk. In conclusion, a depurinized milk diet may be recommended in nutritional treatment of primary and secondary hyperuricemia to avoid uric acid and other volatile, potentially toxic compounds that may slow down liver regeneration and may induce chronic liver diseases.

Determination of styrene content in Gorgonzola PDO cheese by headspace solid phase micro-extraction (HS-SPME) and gas-chromatography mass-spectrometry (GC-MS).

Control of the composition of products that are intended for use as packaging material is essential, particularly when these products come into direct contact with food. It is well known that plastics are not inert and that their residual monomers, starting substances, and additives are able to migrate into the food they contact. Among plastics, styrene is a common compound found in many plastic containers that can also be produced by the oxidation of Penicillium roqueforti used in gorgonzola Protected Denomination of Origin cheese manufacturing. Therefore, solid-phase microextraction combined with gas chromatography/mass spectrometry was applied in the present work to determine the styrene content in packaged and unpackaged gorgonzola cheese samples to understand styrene migration phenomena from plastic containers.

Evidence for false-positive results for boldenone testing of veal urine due to faecal cross-contamination during sampling.

European Directive 96/22/EC, which controls veterinary residues in animals, does not permit the presence of synthetic growth promoters in products of animal origin or in livestock. Boldenone is categorized in class A3 (growth promoters -- steroids) and is thus a banned substance. Testing of veal urine for banned substances is part of the European Union statutory programme for animals going into the food chain. In relation to this monitoring, three studies were conducted to investigate the apparent presence of the banned growth promoter boldenone in veal urine, which was suspected as being caused by interference from faecal contamination of the sample. In the first study, urine samples were collected at different times (time 0 and after 30 min) using (1) a conventional zoonotechnical apron and (2) a technique designed specifically to avoid faecal contamination ('kettle'). This resulted in samples that were, respectively, positive and negative for the presence of alpha-boldenone (alpha-BOL). In a second study, urine samples negative to alpha-BOL were collected from eight veal calves, but became positive after deliberate faecal contamination. In a third study, data obtained from the Italian RNP (Residual National Program) indicated that 18.1% of 3295 urine samples collected using the zootechnical apron were positive for alpha-BOL and 2.1% for beta-boldenone (beta-BOL), whilst of 902 samples collected using the kettle, beta-BOL was not detected in any samples and only 0.2% were positive to alpha-BOL, in concentrations lower than 2 ng ml(-1). These results further support the supposition that faecal contamination of the urine during sample collection can lead to false-positive results during boldenone analysis.

Effects of dietary fats on meat quality and sensory characteristics of heavy pig loins.

This study examined the influence of dietary fat supplementation to heavy pigs on the meat quality and sensory characteristics of loins. The animals were fed diets containing tallow (TA), corn oil (CO) or rapeseed oil (RO) from 25 kg up to 160 kg live weight (LW). The fats were added at 3% as fed from 25 to 110 kg LW, and at 2.5% from 110 kg LW to slaughtering. Diets were fed at 9% BW(0.75). We found no differences between dietary treatments for loin weight, pH, or color of longissimus lumborum (LL) muscle at 45 min and 24 h after slaughtering. No significant differences in moisture, total protein or total lipid content of LL muscle were found. The linolenic acid content of the total lipid of LL muscle was higher in pigs fed RO than those fed TA and CO. The oxidative stability of LL, as determined by induced TBARS, was lower in pigs fed CO - after 60 min of forced oxidation - and in animals fed CO and RO after 300 min, compared to those fed TA. No significant differences in the sensory characteristics of loin were discerned by a panel of trained tasters. These results indicate that long-term nutrition with added fat at the levels we used has little or no effect on the meat quality or sensory characteristics of heavy pig loin.

Gas chromatographic determination of galactose in milk. Example of a switching valve used for the protection of the capillary column.

Galactose, a marker of heat treatment, has been analysed in milk as pentafluorobenzyloxime acetate by gas chromatography with flame ionization detection using a simple switching valve system. The procedure did not entail any prederivatization clean-up for lactose elimination from the sample. In a short pre-column, reagent and lactose derivative excess were separated and the galactose and internal standard derivatives were transferred to the analytical column by a four-port valve. Thus, the analytical column was protected from overloading, so avoiding rapid deterioration.

A new procedure for the specific high-performance liquid chromatographic determination of hydroxyproline.

A procedure suitable for a selective high-performance liquid chromatographic (HPLC) analysis of the imino acid hydroxyproline in the presence of a large excess of amino acids is proposed. To deaminate the amino acids, the well-known reaction with nitrous acid is exploited. The N-nitroso derivatives of imino acids obtained are extracted in ethyl acetate, denitrosated by hydrobromic acid, and treated with dabsyl-chloride. The final HPLC separations are carried out on a reversed-phase column in a rapid isocratic run. The use of the cis isomer of hydroxyproline as an internal standard allows good reproducibility. As an application of the described method, the hydroxyproline content in samples containing collagen and an excess of bovine serum albumine (up to 20:1) is determined.