From the Intersection of Food-Borne Zoonoses and EU Green Policies to an In-Embryo One Health Financial Model

The European Union (EU) adopts the One Health (OH) approach, based on the relationships between human, animal, and environmental health. OH concerns a multitude of aspects, some of which are discussed here. OH overlaps the European Green Deal plan and its relaunched Farm to Fork Strategy, which aims at spreading organic farms adopting the circular economy, in order to improve human health through both better environmental conditions and healthier food. Nevertheless, zoonoses cause sanitary cost in terms of infected farm personnel, lower productivity, and lower fertility of infected farm animals. In such scenarios, the decreased breeding yield and the lower income induce higher cost of farm products, meaning that the market price rises, becoming uncompetitive when compared to the prices of industrial products. Consequently, lower revenues can hinder the farm growth expected in the framework of the EU Green Deal. Since zoonosis control is a key element in aligning EU policies aimed at achieving the EU Green Deal goal of “ZERO environmental impact” by 2050, the authors suggest the inclusion of the parameter economic health in the OH approach, in order to individuate EU Member States (MSs) economically unable to conduct eradication programmes and to finance them. Economic health is here considered as a starting point of the new ethical and science-based One Health Financial Model that the authors suggest as an in-embryo model, in which specific rules should regulate public funds, private investments, and trading, which should exclusively concern public services and private enterprises complying with most of the OH parameters. In this way, economic losses due to collateral negative effects deriving from human activities can be progressively decreased, and the entire planet will benefit from the process. Despite the considerable efforts being carried out in the context of the OH approach, war causes tragic and devastating effects on the physical and mental health of human beings, on their lives, on pandemic and zoonotic threats, on animals, on plants and, last but not least, on the environment. War is incompatible with OH. Enormous efforts for peace are therefore urgently needed

Innovative Caciocavallo cheeses made from a mixture of cow milk with ewe or goat milk.

This study assessed and compared the physicochemical, microbiological, and sensorial characteristics of Caciocavallo cheeses, made from cow milk and a mixture of cow with ewe or goat milk, during ripening. Different cheese-making trials were carried out on an industrial scale following the standard procedure of pasta filata cheeses, with some modifications. The percentage of the different added milk to cow milk influenced compositional and nutritional characteristics of the innovative products, leading to a satisfactory microbiological and sensorial quality

Modulation of the One Health Approach to Tackle Brucellosis in Buffaloes and Cattle in Two Italian Territories with Different Characteristics

The new European Union animal health law and its rule concerning brucellosis in cattle and buffaloes The authors examine the latest European Union (EU) rules concerning eradication and surveillance of brucellosis and animal infectious diseases of EU concern. The Italian rules concerning brucellosis in cattle and buffaloes Italy is included in the EU co-financed compulsory eradication and surveillance programmes for brucellosis in cattle and in buffaloes in the frame of the EU and the related Italian laws, which allowed reaching the Brucellosis Free status without vaccination (former "Officially Brucellosis Free - OBF" status) in the majority of the northern and middle Italian Regions and in some middle and southern Provinces included in Regions where the infection persists. Epidemiology of brucellosis in the EU and in Italy In the Italian Province of Caserta, the highest prevalence of brucellosis in buffaloes is reported; in 2017-2021, a total of 314 outbreaks occurred, in which 39,163 heads tested positive. Here, brucellosis is threatening not only human health and the widespread buffaloes breeding but also the important satellite activities concerning the Protected Designation of Origin (P.O.D.) cheese “Mozzarella di Bufala Campana". The authors also discuss the reemerged brucellosis in cattle in the Molise Region, which despite bordering the Province of Caserta, shows different hydrographic, orographic, and breeding characteristics. In Molise, the reemerged brucellosis had a very different epidemiological course, which allowed it to limit the adoption of One Health measures. The One Health approach to tackle brucellosis in buffaloes In order to tackle brucellosis in Campania Region and in its Province of Caserta, the One Health approach has been predisposed through strict control of animal health, human health, and the environment. The adopted model could be exportable to territories having similar characteristics

Raw milk from vending machines: Effects of boiling, microwave treatment, and refrigeration on microbiological quality

Abstract In Italy, the sale of raw milk from vending machines has been allowed since 2004. Boiling treatment before its use is mandatory for the consumer, because the raw milk could be an important source of foodborne pathogens. This study fits into this context with the aim to evaluate the microbiological quality of 30 raw milk samples periodically collected (March 2013 to July 2013) from 3 vending machines located in Molise, a region of southern Italy. Milk samples were stored for 72h at 4°C and then subjected to different treatments, such as boiling and microwaving, to simulate domestic handling. The results show that all the raw milk samples examined immediately after their collection were affected by high microbial loads, with values very close to or even greater than those acceptable by Italian law. The microbial populations increased during refrigeration, reaching after 72h values of about 8.0 log cfu/mL for Pseudomonas spp., 6.5 log cfu/mL for yeasts, and up to 4.0 log cfu/mL for Enterobacteriaceae . Boiling treatment, applied after 72h to refrigerated milk samples, caused complete decontamination, but negatively affected the nutritional quality of the milk, as demonstrated by a drastic reduction of whey proteins. The microwave treatment at 900 W for 75s produced microbiological decontamination similar to that of boiling, preserving the content in whey proteins of milk. The microbiological characteristics of raw milk observed in this study fully justify the obligation to boil the raw milk from vending machines before consumption. However, this study also showed that domestic boiling causes a drastic reduction in the nutritional value of milk. Microwave treatment could represent a good alternative to boiling, on the condition that the process variables are standardized for safe domestic application

Homology-based modeling of universal stress protein from listeria innocua up-regulated under acid stress conditions

An Universal Stress Protein (USP) expressed under acid stress condition by Listeria innocua ATCC 33090 was investigated. The USP was up-regulated not only in the stationary phase but also during the exponential growth phase. The three dimensional (3D) structure of USP was predicted using a combined proteomic and bioinformatics approach. Phylogenetic analysis showed that the USP from Listeria detected in our study was distant from the USPs of other bacteria (such as Pseudomonas spp., Escherichia coli, Salmonella spp.) and clustered in a separate and heterogeneous class including several USPs from Listeria spp. and Lactobacillus spp. An important information on the studied USP was obtained from the 3D-structure established through the homology modelling procedure. In detail, the Model_USP-691 suggested that the investigated USP had a homo-tetrameric quaternary structure. Each monomer presented an architecture analogous to the Rossmann-like α/β fold with 5 parallel β-strands and 4 α-helices. The analysis of monomer-monomer interfaces and quality of the structure alignments confirmed the model reliability. In fact, the structurally and sequentially conserved hydrophobic residues of the β-strand 5 (in particular the residues V146 and V148) were involved in the inter-chains contact. Moreover, the highly conserved residues I139 and H141 in the region α 4 were involved in the dimer association and functioned as hot spots into monomer-monomer interface assembly. The hypothetical assembly of dimers was also supported by the large interface area and by the negative value of solvation free energy gain upon interface interaction. Finally, the structurally conserved ATP-binding motif G-2X-G-9X-G(S/T-N) suggested for a putative role of ATP in stabilizing the tetrameric assembly of the USP. Therefore, the results obtained from a multiple approach, consisting in the application of kinetic, proteomic, phylogenetic and modelling analyses, suggest that Listeria USP could be considered a new type of ATP-binding USP involved in the response to acid stress condition during the exponential growth phase

Biodiversity of Lactobacillus plantarum from traditional Italian wines

In this study, 23 samples of traditional wines produced in Southern Italy were subjected to microbiological analyses with the aim to identify and biotype the predominant species of lactic acid bacilli. For this purpose, a multiple approach, consisting in the application of both phenotypic (API 50CHL test) and biomolecular methods (polymerase chain reaction-denaturing gradient gel electrophoresis and 16S rRNA gene sequencing) was used. The results showed that Lactobacillus plantarum was the predominant species, whereas Lb. brevis was detected in lower amount. In detail, out of 80 isolates 58 were ascribable to Lb. plantarum and 22 to Lb. brevis. Randomly amplified polymorphic DNA-polymerase chain reaction was used to highlight intraspecific variability among Lb. plantarum strains. Interestingly, the cluster analysis evidenced a relationship between different biotypes of Lb. plantarum and their origin, in terms of wine variety. Data acquired in this work show the possibility to obtain several malolactic fermentation starter cultures, composed by different Lb. plantarum biotypes, for their proper use in winemaking processes which are distinctive for each wine

Detection of Antilisterial Activity of 3-Phenyllactic Acid Using Listeria innocua as a Model

The 3-Phenyllactic acid (PLA) produced by various lactic acid bacteria (LAB) possesses a broad spectrum of antimicrobial activity. In this study, the effect of PLA against Listeria innocua was studied with the aim to obtain additional information about its mechanism of action. The effect of pH on the antilisterial activity of PLA was investigated and a pH-dependent behavior, typical of weak acid, was detected. The antilisterial effect of PLA was firstly compared to that produced by lactic acid (LA) and than to that expressed by phenolic acids (gallic, caffeic, and ferulic acids) evaluating minimum inhibitory concentration (MIC), MBC, and survival kinetic parameters. PLA showed MIC values and death kinetic parameters significantly different from those exhibited by LA and by tested phenolic acids. In particular, the MIC value observed for PLA vs L. innocua resulted lower than that of the other preservative compounds studied herein, and consistent with the quantity generally produced by LAB. Moreover, the effect of PLA and phenolic acids on bacterial surface charge and loss of cellular content resulted different. The overall results highlighted strong differences in the antilisterial mechanism of action among PLA and other compounds such as LA and phenols. Specifically, it is possible to hypothesize that the antilisterial mechanism of action due to PLA is associated with the affinity to cell surface, which contributes to the cellular damage

In Vitro Assessment of Bio-Functional Properties from Lactiplantibacillus plantarum Strains

In recent years, alongside the conventional screening procedures for the evaluation of probiotics for human usage, the pharmaceutical and food industries have encouraged scientific research towards the selection of new probiotic bacterial strains with particular functional features. Therefore, this study intended to explore novel functional properties of five Lactiplantibacillus plantarum strains isolated from bee bread. Specifically, antioxidant, antimicrobial and β-glucosidase activities, exopolysaccharides (EPS) production and the ability to synthesize γ-aminobutyric acid (GABA) were evaluated. The results demonstrated that the investigated L. plantarum strains were effective in inhibiting the growth of some human opportunistic pathogens in vitro (Pseudomonas aeruginosa, Escherichia coli, Proteus mirabilis, Enterococcus faecalis and Staphylococcus aureus). Moreover, the evaluation of antioxidant and β-glucosidase activity and of EPS and GABA production, revealed a different behavior among the strains, testifying how these properties are strongly strain-dependent. This suggests that a careful selection within a given species is important in order to identify appropriate strains for specific biotechnological applications. The results highlighted that the five strains of L. plantarum are promising candidates for application as dietary supplements in the human diet and as microbial cultures in specific food productions