HEMATOLOGY AND BIOCHEMISTRY REFERENCE INTERVALS FOR FREE-LIVING ADULT LESSER KESTRELS (FALCO NAUMANNI) IN SOUTHERN ITALY.

The lesser kestrel (Falco naumanni) is a small falcon with a Euro-Central and Asian-Mediterranean range wintering in sub-Saharan Africa. In the second half of the 20th century, the European population experienced a steep decline and was classified as at risk; thus, its biological and ecological aspects have been widely investigated. Nonetheless, data on hematology and plasma chemistry are not yet available. Therefore, hematology and biochemistry parameters were investigated in a sampling population of clinically healthy lesser kestrels (21 female and 10 male adults) from an Italian rescue center during breeding season, estimating the 95% (2.5 - 97.5th percentile) reference intervals (RI) for standard tests based on either parametric or robust statistical methods. The effect of sex on the referenced parameters was also tested and showed no statically significant differences. The established 95% RI highlighted values comparable with those of other similar raptors such as American kestrel (Falco sparverius) and peregrine falcon (Falco peregrinus). As the first recorded hematology and serum chemistry RI, these clinical data could support conservation efforts and clarify the effects of various environmental and ecological factors on the clinical and health status of lesser kestrels, although they should be reinforced with further data from healthy wild animals.

Rapid Assessment of Metabolomic Fingerprinting of Recycled Sunflower By-Products via DART-HRMS.

To comply with a more circular and environmentally friendly European common agricultural policy, while also valorising sunflower by-products, an ultrasound assisted extraction (UAE) was tested to optimise ethanol-wash solutes (EWS). Furthermore, the capabilities of DART-HRMS as a rapid and cost-effective tool for determining the biochemical changes after valorisation of these defatted sunflower EWS were investigated. Three batches of EWS were doubly processed into optimised EWS (OEWS) samples, which were analysed via DART-HRMS. Then, the metabolic profiles were submitted to a univariate analysis followed by a partial least square discriminant analysis (PLS-DA) allowing the identification of the 15 most informative ions. The assessment of the metabolomic fingerprinting characterising EWS and OEWS resulted in an accurate and well-defined spatial clusterization based on the retrieved pool of informative ions. The outcomes highlighted a significantly higher relative abundance of phenolipid hydroxycinnamoyl-glyceric acid and a lower incidence of free fatty acids and diglycerides due to the ultrasound treatment. These resulting biochemical changes might turn OEWS into a natural antioxidant supplement useful for controlling lipid oxidation and to prolong the shelf-life of foods and feeds. A standardised processing leading to a selective concentration of the desirable bioactive compounds is also advisable.

Discriminative power of DNA-based, volatilome, near infrared spectroscopy, elements and stable isotopes methods for the origin authentication of typical Italian mountain cheese using sPLS-DA modeling.

Origin authentication methods are pivotal in counteracting frauds and provide evidence for certification systems. For these reasons, geographical origin authentication methods are used to ensure product origin. This study focused on the origin authentication (i.e. at the producer level) of a typical mountain cheese origin using various approaches, including shotgun metagenomics, volatilome, near infrared spectroscopy, stable isotopes, and elemental analyses. DNA-based analysis revealed that viral communities achieved a higher classification accuracy rate (97.4 ± 2.6 %) than bacterial communities (96.1 ± 4.0 %). Non-starter lactic acid bacteria and phages specific to each origin were identified. Volatile organic compounds exhibited potential clusters according to cheese origin, with a classification accuracy rate of 90.0 ± 11.1 %. Near-infrared spectroscopy showed lower discriminative power for cheese authentication, yielding only a 76.0 ± 31.6 % classification accuracy rate. Model performances were influenced by specific regions of the infrared spectrum, possibly associated with fat content, lipid profile and protein characteristics. Furthermore, we analyzed the elemental composition of mountain Caciotta cheese and identified significant differences in elements related to dairy equipment, macronutrients, and rare earth elements among different origins. The combination of elements and isotopes showed a decrease in authentication performance (97.0 ± 3.1 %) compared to the original element models, which were found to achieve the best classification accuracy rate (99.0 ± 0.01 %). Overall, our findings emphasize the potential of multi-omics techniques in cheese origin authentication and highlight the complexity of factors influencing cheese composition and hence typicity.

HEMATOLOGY AND BIOCHEMISTRY REFERENCE INTERVALS FOR REHABILITATED EUROPEAN BADGERS (MELES MELES).

The goal of this study was to establish routine reference intervals (RI) for common laboratory tests for the European badger (Meles meles). Blood samples were collected from 13 female and 11 male adult European badgers in a wildlife rehabilitation facility and used for standard hematology and biochemistry analyses. The established 95% RI (2.5th-97.5th percentile) were determined by either parametric (normally distributed data) or robust (nonnormal data) statistical methods and showed a lower range of variability compared with those reported in previous literature. Sex did not affect any investigated parameters. As the first RI study on free-living European badgers in Italy, these reported hematology and serum chemistry RI provide a set of accurate and reliable laboratory parameters for this species.

A Scoping Review on GIS Technologies Applied to Farmed Fish Health Management.

Finfish aquaculture, one of the fastest growing intensive sectors worldwide, is threatened by numerous transmissible diseases that may have devastating impacts on its economic sustainability. This review (2010-2022) used a PRISMA extension for scoping reviews and a text mining approach to explore the extent to which geographical information systems (GIS) are used in farmed fish health management and to unveil the main GIS technologies, databases, and functions used to update the spatiotemporal data underpinning risk and predictive models in aquatic surveillance programmes. After filtering for eligibility criteria, the literature search provided 54 records, highlighting the limited use of GIS technologies for disease prevention and control, as well as the prevalence of GIS application in marine salmonid farming, especially for viruses and parasitic diseases typically associated with these species. The text mining generated five main research areas, underlining a limited range of investigated species, rearing environments, and diseases, as well as highlighting the lack of GIS-based methodologies at the core of such publications. This scoping review provides a source of information for future more detailed literature analyses and outcomes to support the development of geospatial disease spread models and expand in-field GIS technologies for the prevention and mitigation of fish disease epidemics.

Influence of Ageing Time and Method on Beef Quality and Safety.

The effectiveness of dry ageing with regard to retaining meat quality is still subject to debate. At 4 d post mortem, samples of boneless strip loins were excised from young Charolais carcasses and then stored for a further 26 d in a cooler, either vacuum-packaged (VP) or dried-aged (DA). Loin samples were also dissected 7 d post mortem as a control treatment (CT). Chemical, instrumental and microbiological data (n = 18) were determined in longissimus dorsi and underwent ANOVA to estimate the differences in the ageing fixed factor split into two orthogonal contrasts: control vs. aged and VP vs. DA. Ageing loss (both surface dehydration and water purge) was greater in DA compared to VP samples, resulting in the lowest moisture content and highest crude protein and fat percentage in DA loins. The ageing method did not affect meat surface colour, except for redness, which had the lowest value in DA samples. Meat tenderness improved a similar amount following both VP and DA ageing treatments. Compared to the control, prolonged ageing raised both the peroxide value and the total microbial count, especially in DA samples, though both remained within the recommended limits. In summation, both ageing methods improved beef meat tenderisation, preserving its shelf life.

HEMATOLOGIC AND BIOCHEMISTRY REFERENCE INTERVALS FOR RESCUED RED FOXES (VULPES VULPES).

Although the red fox (Vulpes vulpes) is a widespread free-living species in Europe and is often treated as a patient in wildlife rescue centers, peer-reviewed published reference intervals (RI) for hematologic and biochemistry variables are not available. The aim of this study was to determine routine RI for common clinical analyses for this species. After rescue events, single blood samples were collected from 14 female and 18 male adult red foxes and submitted for standard hematologic and biochemistry analyses. The RI were determined by either parametric (normally distributed data) or robust (nonnormal data) statistical methods and showed values close to those reported for specimens of similar fox species, but they were not comparable to historical veterinary clinical data gathered from animals following surgeries or pathology sample collections. Sex did not significantly affect the blood variables, except for iron, which was higher in males. This is the first study reporting RI for a large number of blood analytes in free-living red foxes in Italy. The proposed hematologic and serum chemistry RI, which are specific to red foxes that have recovered after veterinary treatments, represent a set of healthy clinical values that will be helpful for both veterinary care and environmental monitoring.

Seasonal Variation in Raw Milk VOC Profile within Intensive Feeding Systems.

The study aimed to assess the seasonal variation in raw milk volatile organic compounds (VOCs) from three indoor feeding systems based on maize silage (n = 31), silages/hay (n = 19) or hay (n = 16). After headspace solid-phase microextraction (HS-SPME), VOC profiles were determined by gas chromatography (GC). Chemical and VOC (log10 transformations of the peak areas) data were submitted to a two-way ANOVA to assess the feeding system (FS) and season (S) effects; an interactive principal component analysis (iPCA) was also performed. The interaction FS × S was never significant. The FS showed the highest (p < 0.05) protein and casein content for hay-milk samples, while it did not affect any VOCs. Winter milk had higher (p < 0.05) proportions of protein, casein, fat and some carboxylic acids, while summer milk was higher (p < 0.05) in urea and 2-pentanol and methyl aldehydes. The iPCA confirmed a seasonal spatial separation. Carboxylic acids might generate from incomplete esterification in the mammary gland and/or milk lipolytic activity, while aldehydes seemed to be correlated with endogenous lipid or amino acid oxidation and/or feed transfer. The outcomes suggested that VOCs could be an operative support to trace raw milk for further mild processing.

Potential of Full-Fat Silkworm-Based Diets for Laying Quails: Performance and Egg Physical Quality.

The present research was conducted to determine the optimal inclusion level of full-fat silkworm chrysalis meal (SWM) into laying quails' diets, focusing on performance traits and egg physical quality. A total of 240 31-day-old female Japanese quails were randomly assigned to four dietary groups (12 replicates/treatment; 5 quails/replicate); quails were initially fed a standard commercial diet for pullets until 63 days of age. When oviposition started, the experimental groups received the following diets: a conventional corn and soybean-based diet (control diet-C) and three other diets, including 4%, 8%, or 12% of full-fat SWM (SWM4, SWM8, SWM12, respectively). Experimental diets were provided until quails reached 119 days of age. Birds displayed satisfactory productive performance throughout the trial. SWM12 and SWM8 had higher (p < 0.001) egg production but also a higher feed conversion ratio compared to C. At the end of the trial, the eggs edible portion increased, and shell weight decreased with increasing the SWM dietary inclusion level (p < 0.001). At the same time, SWM12 displayed an increase in albumen pH (p < 0.05), even though in the normal range for quail egg. Overall, full-fat SWM (up to 12%) can be considered a promising feed ingredient for laying quails, although higher inclusion levels (>8%) require special attention because SWM also contains anti-nutritional factors.

Ambient mass spectrometry for rapid authentication of milk from Alpine or lowland forage.

Metabolomics approaches, such as direct analysis in real time-high resolution mass spectrometry (DART-HRMS), allow characterising many polar and non-polar compounds useful as authentication biomarkers of dairy chains. By using both a partial least squares discriminant analysis (PLS-DA) and a linear discriminant analysis (LDA), this study aimed to assess the capability of DART-HRMS, coupled with a low-level data fusion, discriminate among milk samples from lowland (silages vs. hay) and Alpine (grazing; APS) systems and identify the most informative biomarkers associated with the main dietary forage. As confirmed also by the LDA performed against the test set, DART-HRMS analysis provided an accurate discrimination of Alpine samples; meanwhile, there was a limited capacity to correctly recognise silage- vs. hay-milks. Supervised multivariate statistics followed by metabolomics hierarchical cluster analysis allowed extrapolating the most significant metabolites. Lowland milk was characterised by a pool of energetic compounds, ketoacid derivates, amines and organic acids. Seven informative DART-HRMS molecular features, mainly monoacylglycerols, could strongly explain the metabolomic variation of Alpine grazing milk and contributed to its classification. The misclassification between the two lowland groups confirmed that the intensive dairy systems would be characterised by a small variation in milk composition.

Multivariate modelling of milk fatty acid profile to discriminate the forages in dairy cows' ration.

Although there are many studies on the importance of fatty acids (FA) in our diet and on the influence of dairy diets on FA metabolism, only a few investigate their predictive capacity to discriminate the type, amount and conservation method of farm forages. This research quantifies differences in milk FA concentrations and, using a supervised factorial discriminant analysis, assesses potential biomarkers when replacing maize with other silages, grass/lucerne hays or fresh grass. The statistical modelling identified three main clusters of milk FA profiles associated with silages, hays and fresh grass as dominant roughages. The main implication of a dairy cow feeding system based on poliphytic forages from permanent meadows is enhancing milk's nutritional quality due to an increase in beneficial omega-3 polyunsaturated FA, conjugated linoleic acids and odd chain FA, compared to feeding maize silage. The study also identified a small but powerful and reliable pool of milk FA that can act as biomarkers to authenticate feeding systems: C16:1 c-9, C17:0, C18:0, C18:3 c-9, c-12, c-15, C18:1 c-9, C18:1 t-11 and C20:0.

Pyrrolizidine alkaloids in bee pollen identified by LC-MS/MS analysis and colour parameters using multivariate class modeling.

Toxic pyrrolizidine alkaloids (PAs) and their N-oxides (PANOs) can be present in bee pollen depending on the plants visited by bees. A liquid chromatography-mass spectrometry (LC-MS/MS) method was developed and validated to monitor 17 PAs/PANOs in 44 bee pollens. The CIE-L∗a∗b∗ colour coordinates with the specular component either included or excluded were recorded in pellets and ground aliquots. Lightness (L∗) and yellowness (b∗) of ground bee pollen were significantly correlated to PAs/PANOs content. The L∗ and b∗ cut-offs sorted by a receiver operating characteristic analysis to predict PAs/PANOs presence showed a significant increase in the relative risk to detect amounts higher than 84 µg kg-1. Two supervised canonical discriminant analyses confirmed that pollen without PAs could be distinguished from those containing PAs/PANOs. The data suggest that instrumental colour coupled with supervised models could be used as a screening test for PAs/PANOs in bee pollen, before the confirmatory LC-MS/MS analysis.

The Effects of Decreasing Dietary Crude Protein on the Growth Performance, Feed Efficiency and Meat Quality of Finishing Charolais Bulls.

This study aimed to evaluate the effect of decreasing dietary crude protein (CP) on the performance of finishing Charolais bulls in the Italian rearing system. Animals were fed two diets, differing only in the CP level (low protein (LP), 13.5% CP versus control (CON), 15.0% CP). Dry matter (DM) intake (DMI) and animals' weights were recorded to obtain average daily gain (ADG) and feed conversion rate (FCR). Feed and fecal samples were collected to evaluate digestibility of diet components. Daily cost of the ration (DRC), feed cost per kg of daily weight gain (CDG) and daily gross margin (DGM) were calculated to analyze the possible benefits of decreasing the protein level. Meat quality analyses were also conducted. Higher DMI (10.6 versus 10 kg/d; p < 0.05) and ADG (1.47 versus 1.36 kg/d; p < 0.05) were observed for CON. No differences in FCR or digestibility were found. Even if the DRC was lower (p < 0.05) for the LP diet (2.26 versus 1.97 €; CON versus LP), no difference was reported for CDG and DGM. Meat lightness and redness were significantly lower and higher in the LP, respectively. To conclude, the CP requirement in these rearing conditions appeared to be higher than 13.5%.

Application of near-infrared spectroscopy for frozen-thawed characterization of cuttlefish (Sepia officinalis).

Few works to date have reported the application of near-infrared spectroscopy (NIRS) for the characterization and authentication of cephalopods. This study investigated the feasibility of a portable NIRS instrument for the non-destructive freshness evaluation of fresh (F) and frozen-thawed (FT) cuttlefish (Sepia officinalis). Samples were examined by chemical, microbiological and sensorial analyses, during 13 days of conservation at 3 °C. The spectral data were collected on lateral mantle of cuttlefish, and different partial least squares discriminant analyses (PLS-DA) were applied for classification purposes. The interpretation of spectra was also investigated by applying the specific water coordinates, using aquaphotomics. Few significant differences in the wet chemistry and microbiological data were detected between F and FT during storage. The quality index method and microbiological analyses suggested similar behavior between F and FT samples until to near 9 days of shelf life. PLS-DA models with the spectral range 900-1650 nm achieved a classification precision of 0.91 between F and FT, while the performances for the prediction of storage days were less effective. The results of aquaphotomics plotted in aquagrams were suitable for the interpretation of the main physicochemical changes of cuttlefish throughout the shelf life. The water coordinates suggested a different molecular conformation of water species in the FT than F samples, with more free water molecules and a lower amount of bound species and the water solvation shell, respectively.

1H NMR Metabolic Profile to Discriminate Pasture Based Alpine Asiago PDO Cheeses.

The study was carried out in an alpine area of North-Eastern Italy to assess the reliability of proton nuclear magnetic resonance 1H NMR to fingerprint and discriminate Asiago PDO cheeses processed in the same dairy plant from upland pasture-based milk or from upland hay-based milk. Six experimental types of Asiago cheese were made from raw milk considering 2 cows' feeding systems (pasture- vs. hay-based milk) and 3 ripening times (2 months, Pressato vs. 4 months, Allevo_4 vs. 6 months, Allevo_6). Samples (n = 55) were submitted to chemical analysis and to 1H NMR coupled with multivariate canonical discriminant analysis. Choline, 2,3-butanediol, lysine, tyrosine, and some signals of sugar-like compounds were suggested as the main water-soluble metabolites useful to discriminate cheese according to cows' feeding system. A wider pool of polar biomarkers explained the variation due to ripening time. The validation procedure based on a predictive set suggested that 1H NMR based metabolomics was an effective fingerprinting tool to identify pasture-based cheese samples with the shortest ripening period (Pressato). The classification to the actual feeding system of more aged cheese samples was less accurate likely due to their chemical and biochemical changes induced by a prolonged maturation process.

Use of rumination and activity data as health status and performance indicators in beef cattle during the early fattening period.

The aim of this study was to measure the level of activity and rumination in young bulls and to assess whether these data can be used as indicators of health status and average daily weight gain (ADG). Two groups of animals (period 1: n=108 animals; period 2: n=106 animals) were fitted with sensors to measure daily activity and rumination, were weighed on arrival and at the end of the trial (70 days) and were checked twice daily to verify their health condition. Any clinical signs and therapies were recorded. The dishomogeneity index of rumination (DR), and the daily dishomogeneity indices of activity (DDA) and rumination (DDR), were calculated. Bulls had an ADG of 1.42±0.38kg/day and showed an average duration of daily rumination of 404±63min and an average activity of 474±46 bits, respectively. Animals characterised by low ADG had lower values of minimum daily rumination (P=0.01) and DDA (P<0.001), and a greater rumination range (P=0.007) and DR (P=0.003). Bovine respiratory disease and lameness were detected 31 and five times, respectively; among affected animals, the average daily activity, rumination and DDA were lower (P<0.05) at 3-6days before the onset of visible clinical signs, whereas DDR increased compared to the values when individuals were apparently healthy. The use of individual sensors appears promising for the early diagnosis of disease in beef cattle and for improving herd management.

Identification of Lipid Biomarkers To Discriminate between the Different Production Systems for Asiago PDO Cheese.

The lipid fraction of Asiago Protected Designation of Origin (PDO) cheese was analyzed to identify specific biomarkers of its main production systems through a canonical discriminant analysis. The three main production systems of the cheese were considered. Two were located in the upland (UL): pasture-based (P-UL) vs hay-based total mixed rations (H-UL). The third was located in the lowland (LL) and processed milk from cows fed maize silage-based rations (maize silage lowland: MS-LL). The discriminant analysis selected nine fatty acids and vitamin A as lipid biomarkers useful to separate the three production systems. High contents of conjugated linoleic acids, anteiso-C15:0, and vitamin A were discriminant factors for P-UL cheese. The separation between H-UL and MS-LL cheese was less marked with the former having the higher content of conjugated linoleic acids and some polyunsaturated n-6 fatty acids and with the latter being identified by cyclopropane fatty acid and C9:0.

Silver As Antibacterial toward Listeria monocytogenes.

Listeria monocytogenes is a serious foodborne pathogen that can contaminate food during processing and can grow during food shelf-life. New types of safe and effective food contact materials embedding antimicrobial agents, like silver, can play an important role in the food industry. The present work aimed at evaluating the in vitro growth kinetics of different strains of L. monocytogenes in the presence of silver, both in its ionic and nano form. The antimicrobial effect was determined by assaying the number of culturable bacterial cells, which formed colonies after incubation in the presence of silver nanoparticles (AgNPs) or silver nitrate (AgNO3). Ionic release experiments were performed in parallel. A different reduction of bacterial viability between silver ionic and nano forms was observed, with a time delayed effect exerted by AgNPs. An association between antimicrobial activity and ions concentration was shown by both silver chemical forms, suggesting the major role of ions in the antimicrobial mode of action.

Metagenomic analysis of rumen microbial population in dairy heifers fed a high grain diet supplemented with dicarboxylic acids or polyphenols.

BACKGROUND: The aim of this study was to investigate the effects of two feed supplements on rumen bacterial communities of heifers fed a high grain diet. Six Holstein-Friesian heifers received one of the following dietary treatments according to a Latin square design: no supplement (control, C), 60 g/day of fumarate-malate (organic acid, O) and 100 g/day of polyphenol-essential oil (P). Rumen fluid was analyzed to assess the microbial population using Illumina sequencing and quantitative real time PCR. RESULTS: The P treatment had the highest number of observed species (P < 0.10), Chao1 index (P < 0.05), abundance based coverage estimated (ACE) (P < 0.05), and Fisher's alpha diversity (P < 0.10). The O treatment had intermediate values between C and P treatments with the exception of the Chao1 index. The PCoA with unweighted Unifrac distance showed a separation among dietary treatments (P = 0.09), above all between the C and P (P = 0.05). The O and P treatments showed a significant increase of the family Christenenellaceae and a decline of Prevotella brevis compared to C. Additionally, the P treatment enhanced the abundance of many taxa belonging to Bacteroidetes, Firmicutes and Tenericutes phyla due to a potential antimicrobial activity of flavonoids that increased competition among bacteria. CONCLUSIONS: Organic acid and polyphenols significantly modified rumen bacterial populations during high-grain feeding in dairy heifers. In particular the polyphenol treatment increased the richness and diversity of rumen microbiota, which are usually high in conditions of physiological rumen pH and rumen function.

Use of dicarboxylic acids and polyphenols to attenuate reticular pH drop and acute phase response in dairy heifers fed a high grain diet.

BACKGROUND: The aim of this study was to determine the ability of two feed additives, a fumarate-malate (FM) and a polyphenol-essential oil mixture (PM), in attenuating the drop of ruminal pH and the metabolic and immune response resulting from an excessively high grain diet. Six heifers were used in a 3 × 3 Latin square experiment and fed a low starch (LS) diet for 14 d, followed by a high starch (HS) diet for 8 d (NDF 33.6%, starch 30.0% DM). In the last 5 days of each period, barley meal was added to decrease rumen pH. During HS feeding all animals were randomly assigned to one of the following three dietary treatments: no supplement/control (CT), a daily dose of 60 g/d of FM, or 100 g/d of PM. Reticular pH was continuously recorded using wireless boluses. On d 21 of each period, rumen fluid was collected by rumenocentesis (1400 h), together with blood (0800 h) and fecal samples (0800, 1400, and 2100 h). RESULTS: The correlation coefficient of pH values obtained using the boluses and rumenocentesis was 0.83. Compared with CT and PM, the FM treatment led to a lower DMI. Nadir pH was lowest during CT (5.40, 5.69, and 5.62 for CT, FM and PM, respectively), confirming the effectiveness of both supplements in reducing the pH drop caused by high grain feeding. This result was confirmed by the highest average time spent daily below 5.6 pH (199, 16 and 18 min/d) and by the highest acetate to propionate ratio of the CT fed heifers. The PM decreased the concentrations of neutrophils (2.9, 3.2, and 2.8 10(9)/L) and acute phase proteins: SAA (37.1, 28.6 and 20.1 µg/mL), LBP (4.1, 3.8, and 2.9 µg/mL), and Hp (675, 695 and 601 µg/mL). Free lipopolysaccharides (LPS) were detected in blood and feces, but their concentrations were not affected by treatments, as the remaining blood variables. CONCLUSIONS: Data suggest that both additives could be useful in attenuating the effects of excessive grain feeding on rumen pH, but the PM supplement was more effective than FM in reducing the inflammatory response compared to CT.