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Simple Summary: Fourier Transform Infrared Spectroscopy (FTIR) is a rapid, cost-effective, and routinely used tool for milk analysis that can be easily applied to the classification of valuable dairy products such as Podolica milk. In the work herein presented, we applied machine learning to rapidly classify the FTIR datasets of milk from different bovine breeds. We were able to successfully recognize non-Podolica milk with 86% sensitivity and 100% specificity, demonstrating that the combination of these tools might be used in the future for the rapid classification of milk from different bovine breeds. The Podolica cattle breed is widespread in southern Italy, and its productivity is characterized by low yields and an extraordinary quality of milk and meats. Most of the milk produced is transformed into "Caciocavallo Podolico" cheese, which is made with 100% Podolica milk. Fourier Transform Infrared Spectroscopy (FTIR) is the technique that, in this research work, was applied together with machine learning to discriminate 100% Podolica milk from contamination of other Calabrian cattle breeds. The analysis on the test set produced a misclassification percentage of 6.7%. Among the 15 non-Podolica samples in the test set, 2 were misclassified and recognized as Podolica milk even though the milk was from other species. The correct classification rate improved to 100% when the same method was applied to the recognition of Podolica and Pezzata Rossa milk produced by the same farm. Furthermore, this technique was tested for the recognition of Podolica milk mixed with milk from other bovine species. The multivariate model and the respective confusion matrices obtained showed that all the 14 Podolica samples (test set) mixed with 40% non-Podolica milk were correctly classified. In addition, Pezzata Rossa milk produced by the same farm was detected as a contaminant in Podolica milk from the same farm down to concentrations as little as 5% with a 100% correct classification rate in the test set. The method described yielded higher accuracy values when applied to the discrimination of milks from different breeds belonging to the same farm. One of the reasons for this phenomenon could be linked to the elimination of the environmental variable. However, the results obtained in this work demonstrate the possibility of using FTIR to discriminate between milks from different breeds. [ABSTRACT FROM AUTHOR]

Simple Summary: The spread of helminths resistant to the most common classes of anthelmintic drugs in sheep and the presence of drug residues in the environment has prompted research to find sustainable alternative solutions for gastrointestinal nematodes control. This study reports the results of in vivo studies on the efficacy of an aqueous Punica granatum-based macerate, used in southern Italy for the control of gastrointestinal nematodes in sheep. The anthelmintic efficacy was evaluated in parallel in sheep infected with gastrointestinal nematodes, using, as a positive control, the treatment with Ivermectin and Albendazole. The results of the study show a good anthelmintic efficacy, suggesting the use of this ethnoveterinary macerate as an alternative and sustainable therapeutical remedy for the helminthiasis control in the sheep. Parasites, in particular, gastrointestinal nematodes (GINs) represent one of the main burdens affecting small ruminant farming and pose a serious threat to their health, welfare, productivity, and reproduction. The correct management of animals and the correct use of anthelmintic drugs are the pillars of the GIN control programs for small ruminants. However, globally due to the indiscriminate use of synthetic anthelmintics, there is a significant increase in anthelmintic resistance phenomena to one or more classes of drugs. Even if such a problem never represented a serious threat in southern Italy because of the favourable environmental conditions and because of the good farm management, the phenomenon is actually showing a steep increasing trend and requires alternative treatment measures and constant monitoring. The use of phytotherapies is considered a valuable alternative approach for GIN control in small ruminants and could help with reducing the amount of synthetic drugs used and the forthcoming anthelmintic resistance. From this perspective, the Calabria territory offers a wide number of plants with anthelmintic efficacy that could be helpful for this purpose. The aim of this study was to evaluate the anthelmintic efficacy of aqueous pomegranate (Punica granatum L.) macerate compared to the treatment with Ivermectin and Albendazole in sheep naturally infected with GINs. The pomegranate macerate derives from the ethnoveterinary knowledge of the Calabria region, Southern Italy. The anthelmintic efficacy was evaluated according to the faecal egg count reduction test (FECRt) using the FLOTAC techniques in two sheep farms in Southern Italy. The FECR was calculated from individual samples using the formula FECR = 100 × (1 − [T2/C2]). The treatment with Albendazole in the first farm showed an efficacy of 99.8% after 14 days and 94.8% after 21 days, while the treatment with Ivermectin in the second farm showed an efficacy of 99.9% after 14 days and 96.5% after 21 days of treatment. The pomegranate macerate, in both farms, showed a value of efficacy of around 50% from day 7 to day 21 after the treatment. Previous studies highlighted the presence of gallic acid as the main component in the pomegranate macerate, and its efficacy in nematode control has been as well previously demonstrated in other plant extracts. This in vivo study demonstrated the unequivocal efficacy of plant macerate in easily reducing 50% of the number of GIN eggs in sheep faeces. These results, obtained without the use of synthetic anthelmintics, indicate the use of green veterinary pharmacology as a sustainable alternative to the use of synthetic drugs to reduce the increase in drug resistance phenomena and the environmental impact. [ABSTRACT FROM AUTHOR]

Simple Summary: This study shows for the first time the variation of the salivary proteome in horses with acute abdominal disease (AAD) compared with healthy horses through a high-throughput proteomic approach. A total of 118 proteins were identified, and 17 showed significant changes between the two groups. The changes observed in proteins were closely related to an impaired primary immune defense and antimicrobial capacity in the mucosa, and one salivary protein (lactoferrin) was successfully verified. These results may increase the background and knowledge of saliva composition in horses with AAD and further understanding of the physiopathological changes occurring in the organism in this disease. The aim of this study was to investigate the changes in the salivary proteome in horses with acute abdominal disease (AAD) using a tandem mass tags (TMT)-based proteomic approach. The saliva samples from eight horses with AAD were compared with six healthy horses in the proteomic study. Additionally, saliva samples from eight horses with AAD and eight controls were used to validate lactoferrin (LF) in saliva. The TMT analysis quantified 118 proteins. Of these, 17 differed significantly between horses with AAD and the healthy controls, 11 being downregulated and 6 upregulated. Our results showed the downregulation of gamma-enteric smooth muscle actin (ACTA2), latherin isoform X1, and LF. These proteins could be closely related to an impaired primary immune defense and antimicrobial capacity in the mucosa. In addition, there was an upregulation of mucin 19 (MUC19) and the serine protease inhibitor Kazal-type 5 (SPINK5) associated with a protective effect during inflammation. The proteins identified in our study could have the potential to be novel biomarkers for diagnosis or monitoring the physiopathology of the disease, especially LF, which decreased in the saliva of horses with AAD and was successfully measured using a commercially available immunoassay. [ABSTRACT FROM AUTHOR]

Simple Summary: Biofilm formation represents one of the most effective forms of bacterial persistence in surfaces where nutrients are available or in the tissues of living hosts as humans or animals. Such persistence is due to the high rate of antimicrobial resistance of this shell conformation. It often represents a burden when the pathogen colonizes niches from where it is not removable such as food facilities, farm facilities or parts of living organisms. In this study, we investigated biofilm formation mechanisms and enhanced antimicrobial resistance of 6 different S. aureus strains. The detected mechanisms were primarily related to the control of catabolites, the production of proteins with moonlighting activities and the detoxification of compounds with antimicrobial activities (i.e., alcohol). Glycolysis and aerobic metabolisms were found to be less active in the biofilm conformation. Consequently, less H2O2 production from aerobic metabolism was translated into a measurable under-representation of catalase protein. Antimicrobial resistance (AMR) represents one of the most critical challenges that humanity will face in the following years. In this context, a "One Health" approach with an integrated multidisciplinary effort involving humans, animals and their surrounding environment is needed to tackle the spread of AMR. One of the most common ways for bacteria to live is to adhere to surfaces and form biofilms. Staphylococcus aureus (S. aureus) can form biofilm on most surfaces and in a wide heterogeneity of environmental conditions. The biofilm guarantees the survival of the S. aureus in harsh environmental conditions and represents an issue for the food industry and animal production. The identification and characterization of biofilm-related proteins may provide interesting insights into biofilm formation mechanisms in S. aureus. In this regard, the aims of this study were: (i) to use proteomics to compare proteomes of S. aureus growing in planktonic and biofilm forms in order to investigate the common features of biofilm formation properties of different strains; (ii) to identify specific biofilm mechanisms that may be involved in AMR. The proteomic analysis showed 14 differentially expressed proteins among biofilm and planktonic forms of S. aureus. Moreover, three proteins, such as alcohol dehydrogenase, ATP-dependent 6-phosphofructokinase, and fructose-bisphosphate aldolase, were only differentially expressed in strains classified as high biofilm producers. Differentially regulated catabolites metabolisms and the switch to lower oxygen-related metabolisms were related to the sessile conformation analyzed. [ABSTRACT FROM AUTHOR]

Simple Summary: Antimicrobials represent useful tools to fight bacterial infections that could harm human and animal health. Antimicrobial resistance occurs naturally or can be induced by the misuse of antibiotics. Its occurrence limits the efficiency of antibiotics and therefore the possibility to treat infections effectively. This can lead to an increasing severity of infectious diseases in humans and animals. Here, we describe the development of a workflow that provides a qualitative representation of the antimicrobial genes that are translated into proteins. Since proteins are ultimately the real effectors, the method herein described demonstrates that those genes are effectively enhancing antimicrobial resistance (AMR). The presented method is independent of any amplification step and provides useful information on the dynamics of the biochemical functions accomplished by the raw milk bacterial consortium. The environment, including animals and animal products, is colonized by bacterial species that are typical and specific of every different ecological niche. Natural and human-related ecological pressure promotes the selection and expression of genes related to antimicrobial resistance (AMR). These genes might be present in a bacterial consortium but might not necessarily be expressed. Their expression could be induced by the presence of antimicrobial compounds that could originate from a given ecological niche or from human activity. In this work, we applied (meta)proteomics analysis of bacterial compartment of raw milk in order to obtain a method that provides a measurement of circulating AMR involved proteins and gathers information about the whole bacterial composition. Results from milk analysis revealed the presence of 29 proteins/proteoforms linked to AMR. The detection of mainly β-lactamases suggests the possibility of using the milk microbiome as a bioindicator for the investigation of AMR. Moreover, it was possible to achieve a culture-free qualitative and functional analysis of raw milk bacterial consortia. [ABSTRACT FROM AUTHOR]

Simple Summary: The treatment of dairy cows with nonsteroidal drugs is applied experimentally to investigate the relevance of inflammation during the periparturient period. Despite appearing healthy, dairy cows throughout the transition period and mainly after parturition can develop a pro-inflammatory status that may negatively influence milk production and cows' health. The administration of nonsteroidal anti-inflammatory drugs (NSAIDs) has been demonstrated to have both positive or negative effects on health and milk production, depending on the type of inhibition mechanism, the dose administered and the cows' lactation numbers. At present, the safety and efficacy of NSAIDs have not been irrefutably demonstrated; therefore, their use to improve metabolic and inflammatory status, as well as milk production and cow health after parturition, should be carefully evaluated. During the transition period, dairy cows experience great physiological stress caused by changes in metabolism and in the immune and endocrine systems. A pro-inflammatory state is another difficulty faced by even apparently healthy animals. The most significant negative consequences of inflammation in dairy cows are substantial impairment of milk production and deleterious effects on cows' health in extreme cases. Nonetheless, a certain degree of inflammation is necessary to sustain physiological adaptations. In recent years, many studies have attempted to determine whether the use of nonsteroidal anti-inflammatory drugs (NSAID) in the transition period of dairy cows could positively affect milk production and cows' health by controlling the inflammation status. This literature indicates that NSAIDs that act as preferential inhibitors of cyclooxygenase-1 (COX-1) activity show important side effects (e.g., increased risk of retained placenta, culling, or metritis) even if milk production is, on average, ameliorated. In contrast, preferential inhibitors of cyclooxygenase-2 (COX-2) activity have overall positive effects on cows' health, with potential beneficial effects on milk production. Furthermore, it is important to note that with certain NSAID treatments, milk discarding is mandatory to prevent contamination with drug residues, but increased milk production can compensate for the loss of milk revenue during the withdrawal period. [ABSTRACT FROM AUTHOR]

Simple Summary: This work described the analysis of differential protein expression of subcutaneous adipose tissue of cows that went under negative energy balance during peripartum. In particular, to the best of our knowledge, it represented an original proteomics study that was able to discriminate cows in negative energy balance up to one month before calving. We believed that our findings would open new perspectives to improve animal welfare during peripartum. To know in advance, the metabolic status of cows would permit to correct the status with appropriate measures, like diet or management. Fat mobilization in high-yielding dairy cows during early lactation occurs to overcome negative energy balance (NEB), caused by insufficient feed intake and the concomitant increased nutritional requirements. For this reason, adipose tissue represents an essential organ for healthy and performant lactation. However, only a few data are known about adipose tissue proteome and its metabolic status during peripartum. The aim of this study was to analyze the differential proteomics profiles of subcutaneous adipose tissue belonging to cows with different NEB scores (low NEB and severe NEB). Both groups were analyzed at three different time points (one month before calving, one and sixteen weeks after calving) that were related to different levels and rates of adipose tissue mobilization. The dataset highlighted the differential expression of the same four key proteins (annexin A2, actin-related protein 10, glyceraldehyde-3-phosphate dehydrogenase, and fatty acid-binding protein) involved in lipid metabolism during all time points and of other 22 proteins typical of the other comparisons among remaining time points. The obtained dataset suggested that the individual variability in adipose tissue metabolism/mobilization/energy availability could be linked to the different outcomes in levels of energy balance and related physical complications among dairy cows during peripartum. [ABSTRACT FROM AUTHOR]