Extensive research has been conducted concerning the determination and characterization of volatile compounds contributing to aroma and flavor in cheese. Considerable knowledge has been accumulated ...on the understanding of the mechanisms through which these compounds are formed during ripening, as well as on the optimization of the methodological approaches which lead to their detection. More recently, particular attention has been given to the aromatic properties of milk and cheeses obtained from lactating dairy ruminants fed experimental diets, characterized, for instance, by the addition of trace elements, natural supplements, or agricultural by-products rich in bioactive compounds. The purpose of this review is to summarize the major families of volatile compounds most commonly found in these types of dairy products at various ripening stages, describing in greater detail the role of animal diet in influencing the synthesis mechanisms most commonly responsible for cheese flavor determination. A large number of volatile compounds, including carboxylic acids, lactones, ketones, alcohols, and aldehydes, can be detected in cheese. The relative percentage of each compound depends on the biochemical processes that occur during ripening, and these are mainly mediated by endogenous enzymes and factors of bacterial origin whose function can be strongly influenced by the bioactive compounds taken by animals with the diet and released in milk through the mammary gland. Further evaluations on the interactions between volatile compounds and cheese matrix would be necessary in order to improve the knowledge on the synthesis mechanisms of such compounds; in addition to this, more should be done with respect to the determination of synergistic effects of flavor compounds, correlating such compounds to the aroma of dairy products.
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Grape pomace (GP) is the main solid by-product of winemaking and represents a rich source of potent bioactive compounds which could display a wide range of beneficial effects in human health for ...their association with reduced risk of several chronic diseases. Several studies have proposed the use of GP as a macro-ingredient to obtain economically worthwhile animal feedstuffs naturally enriched by polyphenols and dietary fibers. Moreover, the research carried out in this field in the last two decades evidences the ability of GP to induce beneficial effects in cow milk and its derived dairy products. First of all, a general increase in concentration of polyunsaturated fatty acids (PUFA) was observed, and this could be considered the reflection of the high content of these compounds in the by-product. Furthermore, an improvement in the oxidative stability of dairy products was observed, presumably as a direct consequence of the high content of bioactive compounds in GP that are credited with high and well-characterized antioxidant functions. Last but not least, particularly in ripened cheeses, volatile compounds (VOCs) were identified, arising both from lipolytic and proteolytic processes and commonly associated with pleasant aromatic notes. In conclusion, the GP introduction in the diet of lactating cows made it possible to obtain dairy products characterized by improved nutritional properties and high health functionality. Furthermore, the presumable improvement of organoleptic properties seems to be effective in contributing to an increase in the consumer acceptability of the novel products. This review aims to evaluate the effect of the dietary GP supplementation on the quality of milk and dairy products deriving from lactating dairy cows.
Preclinical in vivo and in vitro characterization of Antibody-Drug Conjugates (ADCs) involves the development of several bioanalytical methods to address many drug exposure questions. The current ...pharma industry approach requires at least three different assays that must be run, i.e., total antibody (mAb), conjugated payload or conjugated mAb, and free payload assays. Herein we present analytical performances of a quantitative hybrid Ligand Binding/Liquid Chromatography High Resolution and Accuracy Mass Spectrometry (LB/LCHRAM) method that can condense much of the necessary bioanalytical information in one method. The method includes an immuno-capture step, and it detects whole ADC molecules. It was applied to plasma mouse samples and showed reliable bioanalytical performance according to full method validation standards. Quantitation using extracted ion chromatograms and deconvoluted mass peaks was evaluated. The limit of quantitation resulted in 0.5ng of protein on column with a linear dynamic range spanning from 0.5 to 10μg/mL. Moreover, lower drug-to-antibody ratio (DAR) ADC species can be simultaneously detected, also enabling qualitative characterization of in vivo ADC conjugation.
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GEOZS, IJS, IMTLJ, KILJ, KISLJ, NLZOH, NUK, OILJ, PNG, SAZU, SBCE, SBJE, UILJ, UL, UM, UPCLJ, UPUK, ZAGLJ, ZRSKP
Ethical livestock production is currently a major concern for consumers. In parallel, research has shown that transport duration is an important factor affecting animal welfare and has a negative ...impact on the final product quality and on the production cost. This study applied proteomics methods to the animal stress/welfare problem in pigs muscle-exudate with the aim to identify proteins indicative of molecular processes underpinning transport stress and to better characterise this species as a biomedical model. A broader perspective of the problem was obtained by applying label-free LC-MS to characterise the proteome response to transport stress (short or long road transportation) in pigs within the same genetic line. A total of 1,464 proteins were identified, following statistical analysis 66 proteins clearly separating pigs subject to short road transportation and pigs subject long road transportation. These proteins were mainly involved in cellular and metabolic processes. Catalase and stress-induced phosphoprotein-1 were further confirmed by Western blot as being involved in the process of self-protection of the cells in response to stress. This study provide an insight into the molecular processes that are involved in pig adaptability to transport stress and are a step-forward for the development of an objective evaluation method of stress in order to improve animal care and management in farm animals.
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Poultry meat-production is increasing worldwide; leading to the selection of chickens for meat-production that show a fast growth. A label-free quantitative proteomic-approach and Western-blot were ...applied to investigate the dynamics of muscle protein under rapid growth conditions in two common fast-growing broiler genetic-lines (Ross 508 and AZ Extra Heavy Red-chicken). Muscle exudate from chicken Pectoralis major was used as substrate to unveil the proteome of these genetic-lines. Six-hundred forty-five proteins were identified in total from all samples, and after statistical-analysis 172 proteins were found to be differentially-expressed, clearly distinguishing the two chicken genetic-lines. Several of these differentially-expressed proteins were involved with the proteasome and glycolysis/gluconeogenesis-pathways. Changes in meat-quality traits were also observed, which were reflected in the proteomic-profile. Proteins involved in the ubiquitin–proteasome system were associated with the bigger muscle mass of Ross 508, while phosphoglucomutase 1 was associated with a possible higher capability of AZ Extra Heavy Red-chickens to cope with stressors. This pilot proteomic-approach applied on muscle exudate samples provided key evidence about the pathways and processes underlying these two chicken genetic-lines and their meat-quality parameters. We also identified potential biomarkers that could determine the peculiar production potentials (e.g. breast-growth) of these broilers-lines, which arise from differences in their genetic-backgrounds.
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spp. are Gram-negative, non-motile, non-spore-forming, slow-growing, facultative intracellular bacteria causing brucellosis. Brucellosis is an endemic of specific geographic areas and, although ...underreported, represents the most common zoonotic infection, with an annual global incidence of 500,000 cases among humans. Humans represent an occasional host where the infection is mainly caused by
, which is the most virulent;
;
; and
. A microbiological analysis is crucial to identifying human cases because clinical symptoms of human brucellosis are variable and aspecific. The laboratory diagnosis is based on three different microbiological approaches: (i) direct diagnosis by culture, (ii) indirect diagnosis by serological tests, and (iii) direct rapid diagnosis by molecular PCR-based methods. Despite the established experience with serological tests and highly sensitive nucleic acid amplification tests (NAATs), a culture is still considered the "gold standard" in the laboratory diagnosis of brucellosis due to its clinical and epidemiological relevance. Moreover, the automated BC systems now available have increased the sensitivity of BCs and shortened the time to detection of
species. The main limitations of serological tests are the lack of common interpretative criteria, the suboptimal specificity due to interspecies cross-reactivity, and the low sensitivity during the early stage of disease. Despite that, serological tests remain the main diagnostic tool, especially in endemic areas because they are inexpensive, user friendly, and have high negative predictive value. Promising serological tests based on new synthetic antigens have been recently developed together with novel point-of-care tests without the need for dedicated equipment and expertise. NAATs are rapid tests that can help diagnose brucellosis in a few hours with high sensitivity and specificity. Nevertheless, the interpretation of NAAT-positive results requires attention because it may not necessarily indicate an active infection but rather a low bacterial inoculum, DNA from dead bacteria, or a patient that has recovered. Refined NAATs should be developed, and their performances should be compared with those of commercial and home-made molecular tests before being commercialized for the diagnosis of brucellosis. Here, we review and report the most common and updated microbiological diagnostic methods currently available for the laboratory diagnosis of brucellosis.
The Gram‐negative bacteria Brucella ceti and Brucella pinnipedialis circulate in marine environments primarily infecting marine mammals, where they cause an often‐fatal disease named brucellosis. The ...increase of brucellosis among several species of cetaceans and pinnipeds, together with the report of sporadic human infections, raises concerns about the zoonotic potential of these pathogens on a large scale and may pose a threat to coastal communities worldwide. Therefore, the characterization of the B. ceti and B. pinnipedialis genetic features is a priority to better understand the pathological factors that may impact global health. Moreover, an in‐depth functional analysis of the B. ceti and B. pinnipedialis genome in the context of virulence and pathogenesis was not undertaken so far. Within this picture, here we present the comparative whole‐genome characterization of all B. ceti and B. pinnipedialis genomes available in public resources, uncovering a collection of genetic tools possessed by these aquatic bacterial species compared to their zoonotic terrestrial relatives. We show that B. ceti and B. pinnipedialis genomes display a wide host‐range infection capability and a polyphyletic phylogeny within the genus, showing a genomic structure that fits the canonical definition of closeness. Functional genome annotation led to identifying genes related to several pathways involved in mechanisms of infection, others conferring pan‐susceptibility to antimicrobials and a set of virulence genes that highlight the similarity of B. ceti and B. pinnipedialis genotypes to those of Brucella spp. displaying human‐infecting phenotypes.
Orsini and colleagues present the whole‐genome characterization of Brucella ceti and Brucella pinnipedialis, two Gram‐negative bacteria that infect marine mammals causing lethal disease. By de novo assembling of raw sequencing data and comparative annotation of all available genome sequences in databases, the authors provide insights on Brucella genus phylogeny, undertake in‐depth functional analysis of genes involved in virulence and pathogenesis, and highlight the zoonotic potential of these two pathogens that represent a potential threat for the health of coastal communities worldwide.
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The selection and diffusion in the poultry sector of hybrid breeds able to produce higher amount of meat and eggs, led over time to the erosion of genetic resources. One of the strategies that can be ...applied in order to stem such phenomenon, concerns the valorization of the animal products, meat or eggs, obtained from indigenous poultry breeds. Therefore, the aim of this study is to characterize the qualitative aspects of eggs obtained from the Nera Atriana hen, an Italian indigenous laying hen characteristic of the Abruzzo region, making a direct comparison with a commercial hybrid reared in the same environment and applying the same feeding protocol. The trial was conducted for a period of 5 months (from March to July 2019), in which 6 egg samplings were performed (one at the beginning and additional 5 on a monthly basis); each sampling involved the collection of 15 eggs per group of animals for a total of 90 eggs per genotype. Eggs were specifically subjected to evaluations of the physical parameters, including the yolk color, and analyses aimed at determining the content of total lipids, cholesterol, and β-carotene. In addition to this, the fatty acids profile and the electrophoretic pattern of the globular proteins of the yolk were characterized. Eggs from hybrid hens were characterized by higher total weight with significantly higher values specifically associated to albumen and yolk weight and to the albumen percentage. In the eggs obtained from the local hen, higher relative percentages were instead found in the quantity of shell and yolk; furthermore, in the same samples was observed a higher yolk redness, a cholesterol concentration tending to be lower although not significant (
= 0.0521), and a greater amount of saturated fatty acids which could justify a lower susceptibility of the food to oxidative processes following transformation with heat treatments. With regard to the fatty acid composition, differences were specifically observed for palmitic (C16:0) and palmitoleic (C16:1 cis-9) acids that showed higher relative percentages in eggs from local breed, and for the oleic acid (C18:1 cis-9) which was more represented in eggs from commercial hens. In conclusion, some interesting nutritional features, in a preliminary way, have been highlighted that could lead the consumer to prefer these "niche" products over those obtained from commercial hybrids.
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This study aimed to evaluate the effect of dietary integration with extruded linseed (EL) on fatty acid (FA) and aromatic profile of goat cheese after 60 (T60) days of ripening.
Thirty goats were ...divided in two groups. The control group (CG) was fed with conventional diet, whereas the experimental group (EL+) was fed with conventional diet supplemented with 10% of EL. Milk samples were collected on 30 and 60 days of trial to determinate chemical-nutritional composition and FA profile. At the end of experiment, six cheese-making session (3 for each group) were carried out using a pooled milk sample obtained from the 15 goat of each group. At 60 days of ripening, cheeses were analyzed for chemical-nutritional composition, FA and aromatic profile.
An increase in the milk production, protein, fat and lactose were evidenced in the EL+ goats. Conversely, a reduction of somatic cells was observed in the EL+ compared with the CG. However, no variation was observed for urea and casein levels content in milk samples, and no changes in protein and lipid content were found for cheeses in the two experimental groups. Dietary supplementation with EL modified the FA profile of milk. There was a decrease in saturated fatty acids (SFA) and an increase in polyunsaturated fatty acids (PUFA). Chemical composition of T60 cheese did not differ between the two groups but a different FA profile was observed. In particular, in T60 cheese obtained from EL+ milk, an increase in short-chain FA (SCFA) and a decrease in medium and long-chain FA (MCFA and LCFA) were observed. The EL diet led to cheeses with butanoic acid 2 time higher compared to CG cheeses. Moreover, a greater presence of aldehyde compounds and alcohols were observed in the cheeses of experimental group.
The present study pointed out that EL supplementation may improve the chemical and physical qualities of goat milk and cheeses.
The aim of this study was to evaluate the development of volatile compounds in yogurt samples obtained from goats fed a dietary supplementation with olive leaves (OL). For this purpose, thirty Saanen ...goats were divided into two homogeneous groups of 15 goats each: a control group that received a standard diet (CG) and an experimental group whose diet was supplemented with olive leaves (OLG). The trial lasted 28 days, at the end of which the milk of each group was collected and used for yogurt production. Immediately after production, and after 7 days of storage at 4 °C in the absence of light, the yogurt samples were characterized in terms of fatty acid profile, oxidative stability and volatile compounds by the solid-phase microextraction (SPME)-GC/MS technique. Dietary OL supplementation positively affected the fatty acid composition, inducing a significant increase in the relative proportion of unsaturated fatty acids, mainly oleic acid (C18:1
9) and linolenic acid (C18:3). With regard to the volatile profile, both in fresh and yogurt samples stored for 7 days, the OL supplementation induced an increase in free fatty acids, probably due to an increase in lipolysis carried out by microbial and endogenous milk enzymes. Specifically, the largest variations were found for C6, C7, C8 and C10 free fatty acids. In the same samples, a significant decrease in aldehydes, mainly heptanal and nonanal, was also detected, supporting-at least in part-an improvement in the oxidative stability. Moreover, alcohols, esters and ketones appeared lower in OLG samples, while no significant variations were observed for lactones. These findings suggest the positive role of dietary OL supplementation in the production of goats' milk yogurt, with characteristics potentially indicative of an improvement in nutritional properties and flavor.
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