Risk-based scoring and genetic identification for anisakids in frozen fish products from Atlantic FAO areas.

BACKGROUND: The presence of Anisakis larvae in fish represents a major public health concern. Effective risk management procedures should be applied to prevent heavily infected products from reaching the market. The aim of the study is to provide preliminary data on parasite exposure and risk classification in frozen fish products by applying a risk categorization scheme (site, abundance, density and epidemiology - SADE) and Fish Parasite Rating (FPR) method. Fish and cephalopods samples (N = 771) from 5 different FAO Atlantic areas were examined and categorized after an accurate visual inspection and a chloro-peptic digestion. RESULTS: In 25 out of 33 fish species parasite larvae were found. 10897 anisakids larvae were collected and identified to genus level. Molva dypterygia, Conger conger, Zeus faber and Aphanopus carbo were shown to be the most highly infected species. SADE and FPR scores were 1 and poor, respectively, for the referred species, because of the disseminated Anisakis infection and commercial rejection. CONCLUSION: SADE/FPR method showed high specificity and accuracy. The information provided in this work could be used in early warning systems for the detection of parasites in fishery products and might help fishing industries in establishing management strategies for infected stocks in terms of cost saving decisions.

Deltamethrin Residues in Milk and Cheese of Lactating Goats (Capra hircus).

The distribution of pyrethroid insecticide deltamethrin (DLM) in goat milk and cheese (caciotta) following pour-on administration at the sheep dosage (DLMS-10 mL/60 kg body weight) and double dosage (DLMD-20 mL/60 kg body weight) was studied. DLM concentrations were measured in milk collected from study animals (No.14) before treatment and at 2, 4, 8, 12, 16, 24, 30, 36, 48, 56, until 168 h (7 days) post treatment and in caciotta cheese at 12 and 24 h post treatment. At both dosages, the maximum level of DLM residues in goat milk and cheese was below the maximum residue limit (MRL) of 20 µg kg-1 established for bovine milk (EU No 37/2010) at all time points. However, in terms of public health, higher DLM residues in cheese show that further specific studies should be performed on double dosage efficacy and pharmacokinetic and pharmacodynamics properties of ectoparasites in lactating goats.

Examining the Veterinary Electronic Antimicrobial Prescriptions for Dogs and Cats in the Campania Region, Italy: Corrective Strategies Are Imperative.

Companion animals are increasingly being recognised as important contributors to the spread of antimicrobial-resistant bacteria. The present work aimed to measure the antimicrobial drug prescribing in dogs and cats in the Campania Region, Italy by analysing the Veterinary Electronic Prescriptions (VEPs) between 2019 and 2020. The medical records associated with antimicrobial drug prescriptions were collected according to the drug administration (systemic or topical) and the rationale for the treatment chosen. In the period under investigation, 166,879 drugs were prescribed of which 129,116 (73.4%) were antimicrobial. A total of 83,965 (65%) antibiotics were prescribed to dogs, 40,477 (31.4%) to cats, and 4674 (3.6%) to other companion animals. In dogs, 90.5% of VEPs prescribed for systemic treatment included an antimicrobial Critically Important or Highly Important or Important for human medicine (WHO, 2018). The most widely prescribed class was fluoroquinolones. The antimicrobials prescribed were mainly metronidazole-spiramycin (29.7%), amoxicillin-clavulanic (19.6%), enrofloxacin and cephalexin in dogs (16.5%) and enrofloxacin (22.6%) and amoxicillin-clavulanic acid (21.4%) in cats. Based on the results, the widespread use of broad-spectrum antimicrobials and the use of molecules for which limitations should be observed according to the EMA guidelines has emerged.

Data on before and after the Traceability System of Veterinary Antimicrobial Prescriptions in Small Animals at the University Veterinary Teaching Hospital of Naples.

Over recent decades, antimicrobial resistance has been considered one of the most relevant issues of public health. The aim of our study was to evaluate the differences related to the prescription of antimicrobials at the University Veterinary Teaching Hospital, before and after the mandatory use of veterinary electronic prescription (VEP). In particular, the consumption of antimicrobials was examined, especially taking into consideration the recommendations of prudent use. A comparison of data collected before and after the use of electronic prescription highlighted that during the period chosen for the study, the choice of antimicrobial molecules was appropriate, favoring those of "first" and "second line." However, prescription and the use of some molecules not registered for veterinary medicine were observed in the period before VEP. Broad-spectrum antimicrobials, including penicillins with ß-lactamase inhibitors, as well as first-generation cephalosporins and fluoroquinolones, were the most frequently prescribed compounds. There are few studies conducted in Italy aimed at investigating the use of antimicrobials in companion animals under field conditions and with particular regard to prudent use recommendations. This type of study underlines the importance of electronic medical recording in veterinary practice and, above all, its usefulness in monitoring the use of certain antimicrobial agents classified as of critical importance in human medicine.

Hura crepitans L. Extract: Phytochemical Characterization, Antioxidant Activity, and Nanoformulation.

The purpose of this study was to improve the knowledge on Hura crepitans L., a plant belonging to the Euphorbiaceae family that, on the one hand, is known to be toxic, but on the other, is a source of polyphenols with health-promoting effects. Different green extraction methods were applied, varying solvent, temperature, and duration of extraction, which can influence the phytochemical profile and biological activity of plant extracts, and the extracts were fully characterized. Aqueous extracts exhibited a superior antioxidant activity, as indicated by different spectrophotometric tests, and were cytoprotective to HepG2 cells used as model cells. Liquid chromatography-mass spectrometry analyses were performed to identify the secondary metabolites involved in these effects and demonstrated that solvent, duration, and temperature indeed influenced the extraction of polyphenols. Furthermore, the most promising extract, in terms of antioxidant potential, was incorporated into liposomes with the aim of promoting cell interaction and enhancing the antioxidant activity.

Investigation of the Persistence of Penicillin G and Dihydrostreptomycin Residues in Milk of Lactating Buffaloes ( Bubalus bubalis) Using Ultra-High-Performance Liquid Chromatography and Tandem Mass Spectrometry.

The purpose of this research was to evaluate the persistence of penicillin G and dihydrostreptomycin in milk of lactating buffaloes following intramuscular injection of procaine penicillin G (200000 IU/mL) and dihydrostreptomycin sulfate (250 mg/mL) every 24 h for 3 days. Milk samples were collected twice daily up to the 13th milking post-treatment and analyzed by ultra-high-performance liquid chromatography coupled to tandem mass spectrometry. The analytical method has been validated according to Commission Decision 2002/657/EC. The highest concentrations of penicillin G (275 µg kg-1) and dihydrostreptomycin (220.5 µg kg-1) were detected in the milk of the first milkings post-treatment, and levels were below the maximum residue limit of 4 and 200 µg kg-1 in all treated buffaloes at milkings 12 and 2, respectively. The results of this study demonstrate that a nine-milking withdrawal time set for bovine milk was not adequate for depletion of penicillin G in lactating buffaloes.

Evaluation and Comparison of Four Protein Extraction Protocols for Mono- and Two-Dimensional Electrophoresis in Mytilus Galloprovincialis.

In this study, four protein extraction protocols from Mytilus galloprovincialis were evaluated with the aim to identify the most practical, efficient and reproducible method. Four extraction protocols frequently used for mussels and organic matrices were selected and compared. The methods were based on the use of: i) TRIzol reagent; ii) Lysis buffer; iii) phenylmethanesulfonyl fluoride; iv) trichloroacetic acid-acetone. Protein concentration was measured by the Bradford method. Three specimens of mussels were studied and the analysis was conducted in triplicate for each of the four protocols. Results indicated that the four methods could extract significantly different protein profiles. The highest number of protein spots resolved in 2DE gels and the best reproducibility was obtained using trichloroacetic acid-acetone protocol. Results afforded the selection of a suitable extraction protocol to be used for ecotoxicoproteomics studies from mussels and for other proteomic studies conducted by particularly complex tissues such as Mytilus galloprovincialis.

Optimization of Stunning Electrical Parameters to Improve Animal Welfare in a Poultry Slaughterhouse.

Animal killing for food production and the related operations are events that may induce pain, stress, fear and other forms of suffering to the animals. To face this problem and guarantee the animal welfare, the EU has adopted the Regulation (EC) N. 1099/2009 on the protection of animals at the time of killing. Electrical water bath stunning is one of the methods used in commercial slaughterhouses to protect poultry welfare. In particular, this method induces unconsciousness into the birds due to run of electrical current through the head and body. The aim of the present work was to find an optimal setting of electrical parameters to obtain an effective water bath stunning in a commercial poultry slaughterhouse. Moreover, the influence of the tested electrical parameters on meat quality was evaluated. All the experiments confirmed that high stunning frequencies induce a lower occurrence of lesions on carcasses but, on the other hand, require greater current intensities to be effective. A frequency of 750 Hz and an average current intensity of 200 mA for each bird in the water bath resulted as the best combination of electrical parameters to obtain a proper stunning without any consequence on the meat quality.

The Capacity of Listeria Monocytogenes Mutants with In-Frame Deletions in Putative ATP-Binding Cassette Transporters to form Biofilms and Comparison with the Wild Type.

Listeria monocytogenes (Lm) is a food-borne pathogen responsible for human listeriosis, an invasive infection with high mortality rates. Lm has developed efficient strategies for survival under stress conditions such as starvation and wide variations in temperature, pH, and osmolarity. Therefore, Lm can survive in food under multiple stress conditions. Detailed studies to determine the mode of action of this pathogen for survival under stress conditions are important to control Lm in food. It has been shown that genes encoding for ATP-binding cassette (ABC) transporters are induced in Lm in food, in particular under stress conditions. Previous studies showed that these genes are involved in sensitivity to nisin, acids, and salt. The aim of this study was to determine the involvement of some ABC transporters in biofilm formation. Therefore, deletion mutants of ABC transporter genes (LMOf2365_1875 and LMOf2365_1877) were created in Lm F2365, and then were compared to the wild type for their capacity to form biofilms. Lm strain F2365 was chosen as reference since the genome is fully sequenced and furthermore this strain is particularly involved in food-borne outbreaks of listeriosis. Our results showed that ΔLMOf2365_1875 had an increased capacity to form biofilms compared to the wild type, indicating that LMOf2365_1875 negatively regulates biofilm formation. A deeper knowledge on the ability to form biofilms in these mutants may help in the development of intervention strategies to control Lm in food and in the environment.

Persistence of α-cypermethrin residues in milk of lactating donkeys (Equus asinus) using UHPLC-MS/MS.

The aim of this study was to measure the persistence of residues of the pyrethroid insecticide α-cypermethrin (ACYP) in the milk of lactating donkeys following pour-on treatment. Milk was collected from animals (n = 7) before the treatment and at 12, 24, 36, 48, 60, 72 and 84 h post-treatment. The last sampling was taken 7 days post-treatment (168 h). Milk samples were analysed by ultra-high-performance liquid chromatography coupled to tandem mass spectrometry (UHPLC-MS/MS). The analytical method was validated following requirements of Commission Decision 2002/657/EC. All samples showed levels of ACYP below the maximum residue limit (MRL) of 20 µg kg(-1) established for bovine milk (Commission Regulation (EU) No. 37/2010). The results demonstrate that there is minimal partitioning of ACYP into milk in lactating donkeys from pour-on treatment.

Polychlorodibenzodioxin and -furan and dioxin-like polychlorobiphenyl distribution in tissues and dairy products of dairy buffaloes.

A pilot study was performed on three different dairy buffalo herds exposed without exposure control conditions to Polychlorodibenzodioxins and -furans (PCDDs, PCDFs) and Dioxin-like Polychlorobiphenyls (DL-PCBs). This study dealt with the relationship between the contamination levels (pg WHO2005-TE/g fat) in individual raw milk and those in edible tissues and with the contamination transfer from farm bulk milk to dairy products. On a cumulative basis, kidney (41, 67, and 21 pg WHO-TE/g fat) resulted more in equilibrium with milk (48, 42, and 20) than did muscle (25, 31, and 9), while liver showed a large bioaccumulation (221, 304, and 75), with marked differences of the congener profile. Mozzarella cheese contamination (23, 42, and 29 pg WHO-TE/g fat) was higher than that of bulk milk (20, 36, and 21), which suggested a role of casein precipitation in congener transfer. The above information could improve the effectiveness of risk management during a "dioxin" crisis.

Investigation of the persistence of levamisole and oxyclozanide in milk and fate in cheese.

In this study, dairy cows (n = six) were treated with an oral combination product containing levamisole (5 mg/kg body weight, (bw)) and oxyclozanide (10 mg/kg bw). Animals were milked twice daily up to day 16 post-treatment. Milk samples were subsequently analyzed by ultraperformance liquid chromatography coupled to tandem mass spectrometry (UPLC-MS/MS). The highest levels of levamisole (<600 µg/kg) and oxyclozanide (<25 µg/kg) were determined at first and third milking, respectively. Residues of levamisole and oxyclozanide were typically below reporting limits of 0.83 and 1 µg/kg respectively at the 11th and 13th milking, respectively. Soft (3 days ripening), hard (35 days ripening) and whey cheeses were produced from the milk samples collected from the first two milkings. Levamisole residues were found to concentrate in all cheese types. There was a 3-fold concentration effect for levamisole in mature cheese. Oxyclozanide residues were found to occur at lower levels in soft and hard cheese than milk with a 10-fold concentration in whey cheese compared to milk. The results of this study demonstrate that levamisole and oxyclozanide residues are rapidly excreted in dairy cows and milk is compliant after a few days. Oxyclozanide and levamisole residues were shown to be stable during the fermentation process and the whey heat treatment to persist in cheese.