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Economic, social, and environmental sustainability are the results of efforts aiming to improve all aspects of milk production, respecting animal welfare and improving herd health. An epidemiological study was designed to assess the role of contagious pathogens (S. aureus and S. agalactiae) in a cohort of 120 dairy herds located in the southern regions of Italy. Milk quality was assessed using certified methods, and the prevalence of mastitis pathogens in bulk tank milk was determined using quantitative polymerase chain reaction. Welfare scores were assessed using a scoring card that has more than 100 items, including animal-based measurements. Statistical analyses were performed using general lineal model and logistic regression procedures. The results showed that S. aureus had a significant negative effect on the amount of milk nutrients delivered to the dairy plant, and on the level of welfare, whereas the presence of S. agalactiae did not show any significant association. The major risk factors associated with the presence of S. aureus were also identified to help prioritize control programs. These results support the “One Sustainability” approach, implying that an increase in animal productivity is related to the improvement of animal health and welfare and potentially leading to the mitigation of greenhouse gas emissions.

Automatic milking systems (AMSs) are revolutionizing the dairy industry by boosting herd efficiency, primarily through an increased milk yield per cow and reduced labor costs. The performance of milking machines, whether traditional or automated, can be evaluated using advanced vacuum meters through dynamic testing. This process involves scrutinizing the system and milking routine to identify critical points, utilizing the VaDia™ logger (BioControl AS, Rakkestad, Norway). Vacuum recordings were downloaded and analyzed using the VaDia Suite™ software under the guidance of a milking specialist. Access to data from AMSs across various manufacturers and herds facilitated a retrospective study aimed at describing and comparing key milk emission parameters for different AMS brands while identifying potential mastitis risk factors. Using the proper statistical procedures of SPSS 29.1 (IBM Corp., Armonk, NY, USA), researchers analyzed data from 4878 individual quarter milkings from cows in 48 dairy herds. Results indicated a significant variability in milking parameters associated with quarter milk yield and AMS brand. Notably, despite AMSs standardizing teat preparation and stimulation, this study revealed a surprisingly high frequency of two major mastitis risk factors—bimodality and irregular vacuum fluctuations—occurring more frequently than in conventional milking systems. This study, one of the few comparing different AMS brands and their performance, highlights the crucial role of dynamic testing in evaluating AMS performance under real-world conditions.

Several pathotypes of enteric E. coli have been identified. The group represented by Shiga toxin-producing E. coli (STEC) is of particular interest. Raw milk and raw milk products are significant sources of STEC infection in humans; therefore, identifying pathogens at the herd level is crucial for public health. Most national surveillance programs focus solely on raw milk and raw milk cheeses that are ready for retail sale, neglecting the possibility of evaluating the source of contamination directly at the beginning of the dairy chain. To assess the viability of the application of new molecular methodologies to STEC identification in raw milk filters and in calf feces, we analyzed 290 samples from 18 different dairy herds, including 88 bulk tank milk (BTM), 104 raw milk filters (RMF), and 98 calf feces samples. In total 3.4% of BTM, 41.4% of RMF, and 73.4% of calves’ feces were positive for stx, supporting our hypothesis that BTM is not a suitable matrix to assess the presence of STEC at herd level, underestimating it. Our conclusion is that the surveillance program needs critical and extensive improvements such as RMF and calves’ feces analysis implementation to be more efficient in detecting and preventing STEC infections. The epidemiology of these infections and the characteristics of the pathogen clearly show how a One Health approach will be pivotal in improving our capabilities to control the spread of these infections.

Contagious pathogens are very costly to dairy herds, and they may have zoonotic and reverse-zoonotic potentials and may contribute to the spread of antimicrobial resistance. One of the most important risk factors for spreading these infections is milking, when liner contamination may transfer the pathogens from infected to healthy cows. There is no effective protocol to prevent the transmission of infection without the segregation of infected cows. Recently, the availability of elastomers with patented antimicrobial components in their formulations has allowed the exploration of alternative methods to reduce the risk of infection. Two different types of elastomers (rubber and silicone) and nine different formulations were challenged with three major mastitis pathogens (S. aureus, S. agalactiae, and E. coli). The results that were obtained in this study were interesting and unexpected. Indeed, to our knowledge, this is the first study to show that basic rubber materials have intrinsic antimicrobial activity. Silicone elastomers did not exhibit the same levels of bactericidal activity, although they did exhibit some antibacterial capacity. A significant decrease in bacterial survival curves was observed for all the formulations tested when antimicrobial components were added. The different results observed for the various products are likely due to the different formulations and diverse manufacturing processes. The availability of these new materials that significantly reduce the bacterial load on the liner surface may reduce the risk of spreading intramammary infections during milking. This would be an important step forward in achieving global sustainability of dairy herds, consistent with the objectives of One Health, by reducing the risks of zoonotic diseases and antimicrobial treatments.

Staphylococcus aureus are commensal bacteria that are found in food, water, and a variety of settings in addition to being present on the skin and mucosae of both humans and animals. They are regarded as a significant pathogen as well, with a high morbidity that can cause a variety of illnesses. The Centers for Disease Control and Prevention (CDC) has listed them among the most virulent and resistant to antibiotics bacterial pathogens, along with Escherichia coli, Staphylococcus, Klebsiella pneumoniae, Acinetobacter baumannii, Pseudomonas aeruginosa, Enterococcus faecalis, and Enterococcus faecium. Additionally, S. aureus is a part of the global threat posed by the existence of antimicrobial resistance (AMR). Using 26,430 S. aureus isolates from a global public database (NPDIB; NCBI Pathogen Detection Isolate Browser), epidemiological research was conducted. The results corroborate the evidence of notable variations in isolate distribution and ARG (Antimicrobial Resistance Gene) clusters between isolate sources and geographic origins. Furthermore, a link between the isolates from human and animal populations is suggested by the ARG cluster patterns. This result and the widespread dissemination of the pathogens among animal and human populations highlight how crucial it is to learn more about the epidemiology of these antibiotic-resistance-related infections using a One Health approach.

Staphylococcus aureus is considered one of the most widespread bacterial pathogens for both animals and humans, being the causative agent of various diseases like food poisoning, respiratory tract infections, nosocomial bacteremia, and surgical site and cardiovascular infections in humans, as well as clinical and subclinical mastitis, dermatitis, and suppurative infections in animals. Thanks to their genetic flexibility, several virulent and drug-resistant strains have evolved mainly due to horizontal gene transfer and insurgence of point mutations. Infections caused by the colonization of such strains are particularly problematic due to frequently occurring antibiotic resistance, particulary methicillin-resistant S. aureus (MRSA), and are characterized by increased mortality, morbidity, and hospitalization rates compared to those caused by methicillin-sensitive S. aureus (MSSA). S. aureus infections in humans and animals are a prime example of a disease that may be managed by a One Health strategy. In fact, S. aureus is a significant target for control efforts due to its zoonotic potential, the frequency of its illnesses in both humans and animals, and the threat posed by S. aureus antibiotic resistance globally. The results of an epidemiological analysis on a worldwide public database (NCBI Pathogen Detection Isolate Browser; NPDIB) of 35,026 S. aureus isolates were described. We considered the diffusion of antibiotic resistance genes (ARGs), in both human and animal setting, and the results may be considered alarming. The result of this study allowed us to identify the presence of clusters with specific ARG patterns, and that these clusters are associated with different sources of isolation (e.g., human, non-human).

One Health is a well-known strategy for promoting and developing interdisciplinary collaboration across all aspects of health in human, animal, and environmental domains [...]

Clinical and subclinical ketosis (SCK) in dairy cows occurs during the lactation period frequently in many herds, causing a reduction in milk yield and alterations in milk quality with significant economic losses for farmers. SCK is defined as a preclinical stage of ketosis characterized by an elevated ketone body level without clinical signs. Often many cows develop an elevated ketone body level during the first weeks of lactation even though it never goes up to a critical point causing clinical signs. This study aimed to evaluate the prevalence of SCK in Sicily and assess the effect of a treatment with propylene glycol (PG) to control the SCK, thus, reducing the negative effect on milk quality yield. This cross-sectional study was carried out on 22 farms located south-east of Sicily and 1,588 cows in lactation. A total of 3,989 individual milk samples were collected from calving to 80 subsequently days to check the β-hydroxybutyrate (BHB) values in order to establish the SCK status by the Fourier Transform Infrared Spectroscopy. Moreover, the contents of fat, protein, lactose, casein, urea, somatic cell count and acetone were evaluated to identify a correlation between SCK and milk quality. A total of 1,100 cows showed BHB values higher than 0.10 mmol/L. These cows were considered SCK positive, were separated from the rest of the herd, and treated with PG (400 g/head per day), all SCK cows were treated with PG and cows without SCK were not treated. The results showed a prevalence of 41.5% of SCK-positive cows during the first 9 days of lactation. The comparison among the cure rate of treated cows shows that the treatment was most effective in the first 7 days of lactation (76.5% of treated cows) than in the following days. PG positively influenced the milk quality parameters, except for the fat proportion. Moreover, the animals treated with PG showed also an increase in milk yield, supporting the economical sustainability of treatment.

Data on the prevalence of major contagious pathogens in bulk tank milk (BTM) in Italy are generally not available. The availability of Real-Time PCR procedures (qPCR) to perform BTM analysis by represents an important step to define herd health status. Therefore, a cross-sectional epidemiological study was designed to assess the prevalence of contagious pathogens and Prototheca spp in BTM samples. The study was performed on 581 herds from four districts in the west Lombardy region of Italy. Additionally, the relationship between pathogens in BTM and SCC or milk yield; the presence of an association between four risk factors (district, herd size, average milk yield and SCC) with pathogens in BTM were assessed. The overall data showed that S. aureus was recovered in 42% of the herds, Str. agalactiae in 10%, Prototheca spp in 11% and M. bovis in 1.5% of the herds. The GLM model applied showed a significant influence of BTM results, district, herd size and their interactions on SCC and on milk yield variance. Particularly, S. aureus or Str. agalactiae have a significant effect on milk yield variability and, in a lesser extent, on SCC. The very high prevalence of contagious pathogens significantly affects milk characteristics and yield, thus affecting economic sustainability of the herds, and suggests the need to implement control programmes to decrease the prevalence of contagious pathogens, This will also allow to decrease the use of antimicrobials and to improve cow welfare.Highlights First study on a large sample of Italian dairy herds on the prevalence of contagious pathogens in bulk tank milk samples. The prevalence value observed exceeded 50%. First study estimating the prevalence of M. bovis in bulk tank milk in a large sample of Italian dairy herds, and the prevalence observed was 1.5%. Prevalence of contagious pathogens has a significant influence on milk yield and SCC. Bulk tank milk SCC confirmed to have a low accuracy to identify infected herds.

Selective dry cow therapy (SDCT) is one measure suggested to reduce the use of antibiotics on dairy farms. This procedure may have a negative impact on dairy herds, affecting both milk yield and quality. The aim of this observational study was to evaluate the implementation of a SDCT protocol based on the treatment only of cows with SCC at last individual milk test before drying-off >100,000 cells/mL (primiparous cows) or >200,000 cells/mL (pluriparous cows) on udder infection status. We also assessed the association between cow and management factors with the likelihood of cure or of new IMI across the dry period. The study considered 516 dairy cows (2064 quarters), and only 53% of the cows were treated with antibiotics. Before drying-off, 999 quarters (49.1%) were bacteriological negative at two consecutive samples. After calving the negative quarters were 1004 (49.3%). The likelihood of a cure across the dry period was significantly associated with parity, drying-off length and teat sealant application, while the likelihood of new IMI across the dry period was associated to all the risk factors considered (parity, dry period length, teat sealant application, infections status at drying-off and type of long-acting antibiotic applied). The application under field conditions of the proposed SDCT protocol showed to feasible, and to largely reduce the use of antimicrobials at drying-off, with a relatively small impact on cow health.Highlights First observational study in of Italian dairy herds on the application of a selective dry-cow therapy protocol based on SCC. The study showed as a reduction of about 50% in antimicrobial usage at drying off can be achieved. Management and cow factors affect the likelihood of a cure of a new intramammary infection across the dry period.

Mastitis is a major cause of antimicrobial treatments either during lactation or at drying off. From a One Health perspective, there should be a balance between the risk of IMI that may impair cow health and welfare and the reduction of antimicrobial usage to decrease antimicrobial resistance, as may happen when applying selective dry-cow therapy. This reduction may be achieved by an early and accurate diagnosis followed by prudent and rationale therapeutical protocols. This study aims to assess the accuracy of PLCC (neutrophils + lymphocyte count/mL) in identifying cows at risk of having IMI due to major pathogens (S.aureus, Str.agalactiae, Str.uberis, and Str.dysgalactiae), and to simulate the impact of this early diagnosis on the potential number of treatments using a decision-tree model. The results of this study showed that PLCC had an overall accuracy of 77.6%. The results of the decision-tree model based on data from the 12 participating herds, with an overall prevalence of major pathogens of 1.5%, showed a potential decrease in the number of treatments of about 30% (from 3.4% to 2.5%) when PLCC in early lactation (days 5–16) was used to identify cows at risk for major pathogens compared with using SCC at the first milk test (days 17–43). The study confirmed that it is possible to improve animal health and reduce the risk of antimicrobial use through early IMI detection based on PLCC and applying a rationale and prudent antimicrobial protocol.

The aim of this study was to compare the cargos of miRNA in exosomes isolated from the milk of healthy (H) cows, cows at risk of mastitis (ARM), and cows with subclinical mastitis (SCM). Based on the number of somatic cells and the percentage of polymorphonuclear cells, 10 cows were assigned to group H, 11 to group ARM, and 11 to group SCM. After isolating exosomes in milk by isoelectric precipitation and ultracentrifugation, the extracted RNA was sequenced to 50 bp long single reads, and these were mapped against Btau_5.0.1. The resulting 225 miRNAs were uploaded to the miRNet suite, and target genes for Bos taurus were identified based on the miRTarBase and miRanda databases. The list of differentially expressed target genes resulting from the comparisons of the three groups was enriched using the Function Explorer of the Kyoto Encyclopedia of Genes and Genomes. A total of 38, 18, and 12 miRNAs were differentially expressed (DE, p < 0.05) in the comparisons of H vs. ARM, ARM vs. SCM, and H vs. SCM, respectively. Only 1 DE miRNA was shared among the three groups (bta-mir-221), 1 DE miRNA in the H vs. SCM comparison, 9 DE miRNAs in the ARM vs. SCM comparison, and 21 DE miRNAs in the H vs. ARM comparison. A comparison of the enriched pathways of target genes from the H, SCM, and ARM samples showed that 19 pathways were differentially expressed in the three groups, while 56 were expressed in the H vs. SCM comparison and 57 in the H vs. ARM comparison. Analyzing milk exosome miRNA cargos can be considered as a promising approach to study the complex molecular machinery set in motion in response to mastitis in dairy cows.

Milk production loss due to mastitis in dairy herds is economically important. Before estimating the economic impacts of mastitis, it is crucial to quantify the association between mastitis and milk production. The objective of this study was to estimate the association between somatic cell count (SCC, as an indicator of intramammary infection due to mastitis) and milk production for dairy cows in Lombardy, Italy. The test-day (TD) records data of 3816 dairy herds located in three different geographical areas of Lombardy from January 2016 to December 2018 were used. After data editing, the final dataset comprised 10,445,464 TD records from 2970 farms and 826,831 cows. The analysis was carried out by using a mixed-effects model with six fixed effects (geographical Area, Breed, Days in Milk, Parity, Season and Year) and nested random effects for each cow and herd. The results confirmed that the SCC had a negative association with milk production. On average, this study found that any two-fold increase of SCC resulted in a milk production loss of 0.830 (95% CI: −0.832, −0.828) kg/cow/day in the whole of Lombardy. These results can be used for economic calculations on the costs of mastitis.

The application of selective dry cow therapy is one of the measures currently suggested to reduce the use of antibiotics in dairy herds. However, the application of selective dry cow therapy will have a profound impact on Italian dairy herds, very likely affecting both milk yield and quality. Identifying cows to be treated at drying off is crucial for farmers and health authorities, therefore it is necessary the definition of a consistent and certified procedure. This article reports the results of a study aiming to identify which SCC threshold would be the most appropriate to identify cows to be treated and the potential consequences of different selection protocols on udder health after calving under field condition. Last milk test record before drying off and the average of lactation milk test records were considered on a database including 45,682 cow from 709 herd. Five different threshold were considered (50,000; 100,000; 150,000; 200,000; and 250,000 cells/mL). The statistical analysis of the database and a rational evaluation of the results suggest to define thresholds of 100,000 cells/mL for primiparous cows and 200,000 cells/mL for pluriparous cows measured either before drying-off or as the average of all the milk tests of the lactation. The criteria proposed will be useful to manage herd health and, specifically, dry-cows in an efficient and sustainable way, decreasing the use of antimicrobials without increasing the risk of affecting milk yield and quality after calving.Highlights The definition of a consistent and approved procedure to identify cow to be treated in a selective dry cow therapy approach is crucial. SCC from milk test records are a convenient, accurate and certified method. SCC values obtained before drying off or calculated as the average of lactation records can be used. The thresholds of 100,000 cells for primiparous cows and of 200,000 cells for pluriparous cows are suggested as an efficient and sustainable decision tool.

The progressive decrease of mean SCC in dairy herds worldwide is affecting SCC accuracy as a subclinical mastitis marker. This evidence supports studies aiming to apply differential cell count (DSCC) as a tool to identify mastitis. Two of the major obstacles to apply DSCC were the unavailability of high-throughput milk analysers and the cost of these analyses. Recently availability of high-throughput milk analysers, able to perform a partial DSCC on milk, allowed designing a study aiming to identify subclinical mastitis in individual milk samples. This paper reports the result of this first Italian study performed under field conditions. The study considered 4386 milk test records from four dairy herds with different size, management and milking management. DSCC data were analysed by ROC procedure. This procedure allows identifying the threshold giving the highest accuracy and the highest combined value for sensitivity and specificity, among all the possible thresholds. Among the different ways used to classify milk samples, the analysis applied to days in milk (three classes) showed the highest mean values for sensitivity plus specificity, and the value for accuracy was very close to the highest one observed. At the time of submission, this is the first paper available on peer-reviewed scientific journals reporting the evaluation of DSCC as a marker for subclinical mastitis on individual milk samples collected during routine milk test. The results will help the improvement of mastitis diagnosis and will help dairy farmers to increase the levels of herd management and efficiency.Highlights At the time of submission, this is the first paper available on peer-reviewed scientific journals on the evaluation of DSCC as a marker for subclinical mastitis on individual milk samples. The analysis of data showed as DSCC has not consistent performances, confirming the presence of confounding factors such as parity and days in milk. The thresholds calculated on samples classified by days in milk (three classes) showed to have the overall best test performances with an accuracy of 82.3%.

Streptococcus agalactiae is a well-known pathogen in humans and food-producing animals. Therefore, this bacterium is a paradigmatic example of a pathogen to be controlled by a One Health approach. Indeed, the zoonotic and reverse-zoonotic potential of the bacteria, the prevalence of Group B Streptococci (GBS) diseases in both human and animal domains, and the threatening global situation on GBS antibiotic resistance make these bacteria an important target for control programs. An epidemiological analysis using a public database containing sequences of S. agalactiae from all over the world was conducted to evaluate the frequency and evolution of antibiotic resistance genes in those isolates. The database we considered (NCBI pathogen detection isolate browser—NPDIB) is maintained on a voluntary basis. Therefore, it does not follow strict epidemiological criteria. However, it may be considered representative of the bacterial population related to human diseases. The results showed that the number of reported sequences increased largely in the last four years, and about 50% are of European origin. The frequency data and the cluster analysis showed that the AMR genes increased in frequency in recent years and suggest the importance of verifying the application of prudent protocols for antimicrobials in areas with an increasing frequency of GBS infections both in human and veterinary medicine.

Mastitis is one of the major diseases of dairy cows that affects milk quality and quantity and increases the potential risk for the presence of antimicrobial residues (AR) in milk, which could lead to the development of antimicrobial resistance (AMR) among human pathogens. Even if the presence of AR in milk and milk products is low in many countries, the threat is not negligible and cannot be ignored. These problems may be investigated by applying a One Health approach, and this prevalence study aimed to estimate the risks for human health related to milk production applied to dairy herds in Lombardy. Three hundred thirty-one bulk tank milk samples were randomly collected and analyzed by CombiFoss 7 and MilkoScan 7 (milk quality, bacteria, and somatic cell count), an HPLC system coupled to a Q-Exactive Orbitrap (AR), and qPCR (contagious pathogens). The data were analyzed by a generalized linear model. The results showed a relatively high prevalence of contagious pathogens (S. aureus 28.1%; Str. agalactiae 7.3%; M. bovis 3%), which primarily affect milk nutritional components decreasing mainly milk fat content (range 1%–2.5%), but did not show them to be associated to an increase of the risk of antimicrobial residues. These latter ones were recovered only in 7/331 samples at concentrations far below official MLRs. The results support currently active surveillance programs’ efficacy in reducing AR risks, which may be further improved by prioritizing them based on geographical area characteristics.

The factors that influence the diversity and composition of raw milk and fecal microbiota in healthy commercial dairy herds are not fully understood, partially because the majority of metataxonomic studies involve experimental farms and/or single factors. We analyzed the raw milk and fecal microbiota of 100 healthy cows from 10 commercial alpine farms from the Province of Trento, Italy, using metataxonomics and applied statistical modelling to investigate which extrinsic and intrinsic parameters (e.g. herd, diet and milk characteristics) correlated with microbiota richness and composition in these relatively small traditional farms. We confirmed that Firmicutes, Ruminococcaceae and Lachnospiraceae families dominated the fecal and milk samples of these dairy cows, but in addition, we found an association between the number of observed OTUs and Shannon entropy on each farm that indicates higher microbiota richness is associated with increased microbiota stability. Modelling showed that herd was the most significant factor affecting the variation in both milk and fecal microbiota composition. Furthermore, the most important predictors explaining the variation of microbiota richness were milk characteristics (i.e. percentage fat) and diet for milk and fecal samples, respectively. We discuss how high intra-herd variation could affect the development of treatments based on microbiota manipulation.

One critical point of dairy farm management is calving and neonatal first care. Timely calving assistance is associated with the reduction of calf mortality and postpartum uterine disease, and with improved fertility in dairy cattle. This study aimed to evaluate the performance and profitability of an intravaginal sensor for the prediction of stage II of labor in dairy farms, thus allowing proper calving assistance. Seventy-three late-gestating Italian Holstein cows were submitted to the insertion of an intravaginal device, equipped with light and temperature sensors, connected with a Central Unit for the commutation of a radio-signal into a cell phone alert. The remote calving alarm correctly identified the beginning of the expulsive phase of labor in 86.3% of the monitored cows. The mean interval from alarm to complete expulsion of the fetus was 71.56 ± 52.98 min, with a greater range in cows with dystocia (p = 0.012). The sensor worked correctly in both cold and warm weather conditions, and during day- or night-time. The intravaginal probe was well tolerated, as any cow showed lesions to the vaginal mucosa after calving. Using sex-sorted semen in heifers and beef bull semen in cows at their last lactation, the economic estimation performed through PrecisionTree™ software led to an income improvement of 119 € and 123 €/monitored delivery in primiparous and pluriparous cows, respectively. Remote calving alarm devices are key components of “precision farming” management and proven to improve animal welfare, to reduce calf losses and to increase farm incomes.

The One Health approach emphasizes the importance of antimicrobial resistance (AMR) as a major concern both in public health and in food animal production systems. As a general classification, E. coli can be distinguished based on the ability to cause infection of the gastrointestinal system (IPEC) or outside of it (ExPEC). Among the different pathogens, E. coli are becoming of great importance, and it has been suggested that ExPEC may harbor resistance genes that may be transferred to pathogenic or opportunistic bacteria. ExPEC strains are versatile bacteria that can cause urinary tract, bloodstream, prostate, and other infections at non-intestinal sites. In this context of rapidly increasing multidrug-resistance worldwide and a diminishingly effective antimicrobial arsenal to tackle resistant strains. ExPEC infections are now a serious public health threat worldwide. However, the clinical and economic impact of these infections and their optimal management are challenging, and consequently, there is an increasing awareness of the importance of ExPECs amongst healthcare professionals and the general public alike. This review aims to describe pathotype characteristics of ExPEC to increase our knowledge of these bacteria and, consequently, to increase our chances to control them and reduce the risk for AMR, following a One Health approach.

There is increasing evidence to suggest that, in addition to their regenerative effect, mesenchymal stromal cells (MSCs), and their secretome have an anti-inflammatory and antimicrobial role in the innate immune response in conditions such as sepsis. However, there is no published information on the effect of MSCs in bovine mastitis. Mastitis often results in extensive tissue damage due to multi-microorganism co-infection. This study investigated the ability of amniotic-derived conditioned medium (CM), in vitro and in vivo, to counteract microbial action and restore healthy tissue capable of milk production. Following determination of a dose–response curve, 10,000 colony-forming units (CFU) of Staphylococcus aureus (S. aureus) were inoculated into bovine mammary epithelial cell culture with and without 10% CM (supplemented either at the time of bacteria inoculation or after 4 h). Acridine orange staining was used to assess cell viability/apoptosis. Additionally, an in vivo study was performed using 48 dairy cows with acute and chronic mastitis, treated with CM (treated group) or antibiotics (control group). In vitro results showed that CM can attenuate bacterial growth, as evaluated by the number of CFU. After 24 h of culture with S. aureus, 89.67% of mammary epithelial cells treated with CM were still alive, whereas all cells cultured without CM were dead. Rates of epithelial cell survival (60.67%) were similar when CM was added 4 h after bacteria inoculation. There was no difference in somatic cell count between cases of acute mastitis in the CM-treated or control group in the in vivo study. However, relapses in chronic mastitis were less common in the group receiving CM. Our results show that CM is able to mitigate bacterial growth in vitro and may be particularly useful in the treatment of chronic mastitis, aiding restoration of milk production in cows that would otherwise be removed from the production cycle.

In this study we developed a model for risk prioritisation and characterisation focused on zoonoses and food safety for diseases of interest in veterinary public health at a regional level in Italy. A previous model (Discontools) based on scorecards was used as a basis to develop the new model. A Formalised Consensus Process approach involving academics and veterinary officers was used to develop scorecards and relative form and guidelines. Scorecards include several areas of interest, with different categories and coefficient of importance. The following areas were identified: relevance of the disease, socio-economic impact, impact on public health, impact on trade, impact on animal welfare, control tools. A guide and a form were finalised in order to fill scorecards. Scorecards were filled by consulting available data, literature, and expert opinions. Among bovine diseases, mastitis (Salmonella aureus) showed the highest score; Q fever was the highest among small ruminants; among swine diseases the highest was salmonellosis; while among other animal diseases, toxoplasmosis had the highest score. The approach described in this study is designed to aid professionals in risk prioritisation, decision-making, and to improve disease control systems at a regional level in Italy. It also facilitates risk characterisation in different backgrounds and the identification of data holes in specific areas of interest for the diseases considered. This approach is conceived to aid professionals in risk prioritization, decision-making and to improve disease control systems at a regional level. It also allows to perform risk characterization in different backgrounds and to identify lacks of data in specific areas of interest for the diseases considered.

Mastitis is an inflammatory disease of the mammary gland, caused by the invasion of microorganism on this site, associated with an altered immune response. Recent studies in this field hypothesize that the origin of these pathogens can also be from the gastrointestinal tract, through the entero-mammary pathway in relation to an increase in gut permeability. In this study, we wanted to investigate if inflammatory status of the mammary gland is related to an alteration of gut permeability. The microbiome of feces, blood and milk of lactating cows, recruited on the basis of the total somatic cell count and of the percentage of polymorphonuclear neutrophils and lymphocytes, was studied. Cows were divided into healthy (G), at risk of mastitis (Y) and with mastitis (R) classifications. The bacterial DNA was extracted and the V3 and V4 regions of 16S rRNA sequenced. Moreover, the quantification of total bacteria was performed with quantitative real-time PCR. A non-parametric Kruskal–Wallis test was applied at the phylum, family and genera levels and beta biodiversity was evaluated with the unweighted UniFrac distance metric. Significant differences between groups were found for the microbial composition of feces (Clostridiaceae, Turicibacteriaceae for family level and Clostridium, Dorea, SMB53 and Turicibacter for genus level), blood (Tenericutes for phylum level and Mycoplasma for genus level) and milk (OD1 and Proteobacteria for phylum level, Enterobacteriaceae and Moraxallaceae for family level and Olsenella and Rhodococcus for genus level). The beta biodiversity of feces and blood did not change between groups. Significant differences (p < 0.05) were observed between the beta diversity in milk of G group and Y group and between Y group and R group. The number of taxa in common between feces, blood and milk were 8 at a phylum, 19 at a family and 15 at a genus level. From these results, the bacterial crossing from gut to milk in cows was not confirmed but remained hypothetical and deserves further investigation.

The virus responsible for the pandemic that has affected 152 countries worldwide is a new strain of coronavirus (CoV), which belongs to a family of viruses widespread in many animal species, including birds, and mammals including humans. Indeed, CoVs are known in veterinary medicine affecting several species, and causing respiratory and/or enteric, systemic diseases and reproductive disease in poultry. Animal diseases caused by CoV may be considered from the following different perspectives: livestock and poultry CoVs cause mainly “population disease”; while in companion animals they are a source of mainly “individual/single subject disease”. Therefore, respiratory CoV diseases in high-density, large populations of livestock or poultry may be a suitable example for the current SARS-CoV-2/COVID-19 pandemic. In this review we describe some strategies applied in veterinary medicine to control CoV and discuss if they may help to develop practical and useful strategies to control the SARS-CoV-2/COVID-19 pandemic.

Milk differential somatic cells count (DSCC), made possible under field conditions by the recent availability of a high-throughput milk analyzer may represent an improvement in mastitis diagnosis. While an increasing number of studies reports data on DSCC on individual cow samples, very few concerns DSCC from quarter milk samples. This paper reports for the first time the results of a retrospective study aiming to assess the performance of total (SCC), DSCC, and a novel calculated marker (PLCC) measured on quarter milk samples as a method to identify cows at risk for intramammary infection (IMI) in the first 30 days after calving. Overall, 14,586 valid quarter milk samples (3658 cows) taken in the first 30 days of lactation were considered. Quarters with major pathogens (MP) IMI, as expected, showed significantly higher means for SCC, DSCC, and PLCC. The accuracy, sensitivity, and specificity of the diagnosis based on different cut-offs calculated by ROC analysis are relatively close among DSCC, PLCC, and SCC (up to cut-off of 200,000 cells/mL). However, decision-tree analysis which includes the costs of analysis, but also the costs of the actions taken after test results showed as PLCC has the lowest cost among the three markers, and PLCC and SCC are cost effective when MP prevalence is higher than 6–10%. This diagnostic approach is of high interest particularly when selective dry cow therapy is applied to improve animal health at the herd level.

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.

Differential leukocyte count (DSCC) in milk is considered important to improve knowledge of udder immune response. The investigations on milk DSCC were limited by the techniques available until recently, when a high-throughput tool to perform DSCC opened the way to explore these factors in rapid and economically sustainable ways. We hypothesized that DSCC alone does not fully describe the pattern of these cells, since the total amount is also influenced by milk yield and SCC. Therefore, this study was designed to describe DSCC and total amount of different leukocytes in milk during the course of lactation in cows differing in parity and in levels of SCC. This study considered 17,939 individual milk tests from 12 dairy herds in Lombardy Region, where DCC testing was applied in the period of February 2018–December 2019 (23 months). The samples were divided into two subsets—“healthy” (HS) with SCC ≤200,000 cells/mL and “inflamed” (IS) with SCC >200,000 cells/mL. Cow in HS have a P + LT average between 5.0 × 108 and 3.0 × 109 cells. In IS cows, the values were 1.6 × 1010 and 2.5 × 1010. Therefore, the presence of a well-defined inflammatory process increased the overall amount of polymorphonuclear neutrophils (PMN) and lymphocytes (LYM) of 1 log, from 1 × 109 to 1 × 1010. The assessment of the total amount of PMN and LYM, to our knowledge, have never been reported in scientific literature; the values observed may be proposed as benchmarks for studies on udder immune response. When data were analyzed by days in milk (DIM), they showed that cows in first and second lactation have a significantly lower amount of PMN + LYM, when compared to cows in third and higher lactation. However, these differences are numerically not very large (7%), and suggest that, in healthy animals, the number of immune cells is kept as constant as possible. In IS, the analysis of trends based on DIM showed that both DSCC and P + LT have a significant negative trend. These data suggest that only in this group, the presence of high SCC as lactation proceeds is associated with a progressive increase in the number of macrophages. To the best of our knowledge, this is the first study describing the pattern of DSCC and the total amount of PMN + LYM in relation to parity, days in milk, and SCC, and it may be considered as the first contribution in the investigation on mammary gland immune response by the means of differential cell counts in milk.

Recent available instruments allow to record the number of differential somatic cell count (DSCC), representing the combined proportion of polymorphonuclear leukocytes and lymphocytes, on a large number of milk samples. Milk DSCC provides indirect information on the udder health status of dairy cows. However, literature is limited regarding the effect of DSCC on milk composition at the individual cow level, as well as its relation to the somatic cell score (SCS). Hence, the aims of this study were to (i) investigate the effect of different levels of DSCC on milk composition (fat, protein, casein, casein index, and lactose) and (ii) explore the combined effect of DSCC and SCS on these traits. Statistical models included the fixed effects of days in milk, parity, SCS, DSCC and the interaction between SCS × DSCC, and the random effects of herd, animal within parity, and repeated measurements within cow. Results evidenced a decrease of milk fat and an increase in milk fatty acids at increasing DSCC levels, while protein, casein and their proportion showed their lowest values at the highest DSCC. A positive association was found between DSCC and lactose. The interaction between SCS and DSCC was important for lactose and casein index, as they varied differently upon high and low SCS and according to DSCC levels.

The recent availability of a high-throughput milk analyzer performing a partial differential somatic cell count (DSCC) opened new opportunities in investigations on bovine udder health. This analyzer has a potential limitation on the accuracy of measurements when the somatic cell count (SCC) is below 50,000 cells/mL, values characterizing a good proportion of lactating cows in many herds. We obtained data for cows below this threshold, assessed the repeatability of these measurements and investigated the relationship between DSCC and udder health, milk composition and yield. Overall, 3022 cow milk test records performed on a Fossomatic™ 7/DC (Foss A/S, Hillerød, Denmark) were considered; 901 of them had an SCC ≤ 50,000 cells/mL. These latter samples were analyzed by qPCR to identify the presence of bacteria. Overall, 20.75% of the samples (187) were positive. However, the health status did not have any significant association with DSCC. The analysis of the association of DSCC on milk fat, protein and casein showed a significant decrease in their proportions as the DSCC increased, whereas it was not observed for milk yield and lactose. Therefore, DSCC in very low SCC cows may be suggested as a marker to identify early changes in milk composition.

Despite the fact that health communication is a discipline developed only recently, its importance in human medicine is well recognized. However, it is less considered in veterinary medicine, even if it has the potential to improve public health because of the role of veterinary medicine in public health. For this reason, an One Health approach is useful for communication as well. This approach leads to a “One Communication” concept, which is the result of the synergy in communicative efforts both in human and in veterinary medicine. Our analysis explores the potential of communication in several veterinary fields: institutions, food safety, companion animal and food-producing animal practice, pharmacology and drugs, wildlife fauna and environment. In almost all the areas of veterinary activity communication can contribute to human health. It takes many forms and use several channels, and this variety of communicative opportunities represent a challenge for veterinarians. For this reason, the communication course should be included in the curricula of Veterinary Medicine Schools. As One Health, One Communication is a strategy for expanding collaborations in health communication and it will enhance public health.

Nowadays food wrapping assures attractive presentation and simplifies self-service shopping. Polyvinylchloride (PVC)- and polyethylene (PE)-based cling-films are widely used worldwide for wrapping cheeses. For this purpose, films used in retail possess suitable technical properties such as clinginess and unrolling capacity, that are achieved by using specific plasticizers during their manufacturing process. In the present study, the main VOCs of three cling-films (either PVC-based or PE-based) for retail use were characterized by means of Solid-Phase Micro-Extraction and GC/MS. In addition, the effects of cling film type and contact time on the migration of VOCs from the films to four different PDO Italian cheeses during cold storage under light or dark were also investigated. Among the VOCs isolated from cling-films, PVC released 2-ethylhexanol and triacetin. These compounds can likely be considered as a “non-intentionally added substance”. These same compounds were also detected in cheeses wrapped in PVC films with the highest concentration found after 20 days storage. The PE cling-film was shown to possess a simpler VOC profile, lacking some molecules peculiar to PVC films. The same conclusions can be drawn for cheeses wrapped in the PE cling-film. Other VOCs found in wrapped cheeses were likely to have been released either by direct transfer from the materials used for the manufacture of cling-films or from contamination of the films. Overall, HS-SPME is shown to be a rapid and solvent free technique to screen the VOCs profile of cling-films, and to detect VOCs migration from cling-films to cheese under real retail storage conditions.

The aim of this research was to evaluate the effect of the modification of diet starch content on milk composition and onits nitrogen and mineral fractions. Ten Italian Friesian primiparous cows were randomly assigned to two groups and feda basal total mixed ration, (BSD, basal starch diet, 24.9% starch/DM) until 42 days in milking (DIM). At 43 DIM, 5 animals(control group, CTR) continued to receive the same ration and the remaining 5 cows (experimental group, EXP)were fed a low starch diet (LSD, 21.0% starch/DM) until 65 DIM, followed by a high starch diet (HSD, 28.3% starch/DM)66 to 85 DIM. From 86 DIM until 94 DIM, cows of the EXP group returned to the BSD. Milk samples were collected at 37,50, 60, 70, 80, 94 DIM. Starch intake was lower for EXP at DIM 50, 60 and higher at 70 and 80 DIM (P < 0.01). Milkyield and fat corrected milk (FCM) did not vary between groups and times of sampling, but HSD caused a significant (P< 0.05) reduction of milk fat in the EXP animals. NPN and MUN contents were affected by dietary treatments at 80 DIM(P < 0.05), being higher in CTR and lower in EXP group in comparison to their basal values (37 DIM). The αs1-caseinpercentages at 80 and 94 DIM increased in the EXP but not in the CTR group, and a marked decrease of αs2-casein percentagefor the EXP group at 94 DIM was observed (P < 0.05). Milk sodium content decreased at 80 and 94 DIM and thesodium to potassium ratio was reduced after the return to the basal diet in the EXP group (94 DIM), indicating that dietarystarch variations can be involved in the control of epithelium integrity of mammary gland in early lactation.

To assess the impact of automatic milking systems (AMS) on the different aspects of milk production a research projectinvolving both commercial and experimental dairy farms with different AMS and different management was started. Thispaper reports the results of a follow-up study on primiparous cows focused on assessing some markers to be used tomonitor udder and teat health. Heifers were included after calving and sampled for at least 12 months. Quarter milk samplesand teat measurements were taken to assess: intramammary infection (IMI) frequencies, somatic cell counts (SCC),teat thickness changes, teat skin and apex conditions. The study included 28 cows in herd A and 27 in herd B for a totalnumber of 2344 samples. Overall, teat apex and skin conditions were maintained along the lactation. Teat skin conditionstended to decrease because of the accumulated number of milkings while lactation proceeded, but at a largelyacceptable level in both herds. Teat apex conditions showed a decrease. Teat thickness changes displayed different patternsin the two herds, probably because of the different type of AMS, but in both cases a trend to decrease in thicknesscould be observed. The application of AMS in herd B, free from contagious pathogens, did not influence the frequency ofIMI and the SCC. In herd A, characterized by the presence of Staphylococcus aureus IMI, the frequency of IMI showeda progressive increase, very likely as a consequence of the spread of infections during milking. Teat skin had no associationwith the frequency of IMI. Teat thickness changes outside values considered as physiological proved to be associatedwith decreased conditions in the teat apex score in herd A, but not in herd B. However, a decrease in teat apex scoreproved to be associated with an increase in IMI frequency in both herds. The results of this field trial confirm that AMShave no negative impact on IMI incidence, SCC and teat tissue conditions when the initial cow health status and overallherd management are good. In the presence of contagious pathogens, and when cows have more than 300 days in milk(DIM) the frequency of negative outcomes significantly increases and the proper control measures should be taken.

The aim of the study was to investigate the relationships between milk flow traits, milk Somatic Cell Count (SCC) and teat apex score in primiparous cows during the first 120 days of lactation. A total of 82 primiparous Holstein cows were randomly selected in 6 herds characterized by different milking machines and management. The cows were monitored monthly during the first 120 days of lactation. Each month, milk flow curves of the whole udder of each primiparous cow were registered with continuous electronic milk flow meters (Lactocorder), quarter milk samples were aseptically collected, somatic cells were counted, and teat apex scores were assessed. Quarter milk samples were divided in two groups according to SCC: 200,000 cells/ml, respectively. At udder level each cow was classified as low SCC (LSCC) when all quarters had