Immunomodulatory Role of Rosmarinus officinalis L., Mentha x piperita L., and Lavandula angustifolia L. Essential Oils in Sheep Peripheral Blood Mononuclear Cells.

Recently, the uses of essential oils (EOs) as rumen modifiers, anti-inflammatory agents, and antioxidants were demonstrated in livestock. In the present study, the role of Mentha x piperita L. (MEO), Rosmarinus officinalis L. (REO), and Lavandula angustifolia L. (LEO) EOs in an in vitro sheep model of inflammation was investigated. With this aim, peripheral blood mononuclear cells (PBMCs) were treated with incremental concentrations (3, 5, 7, and 10%) of each EO to test their effects on cell viability and proliferation and on interleukin (IL)-6, IL-10, and IL-8 secretion. The PBMCs were stimulated by Concanavalin A (ConA) alone or in combination with lipopolysaccharide (LPS) mitogen. The positive and negative controls were represented by PBMCs in the presence or absence, respectively, of mitogens only. The cell viability and proliferation were determined by XTT and BrdU assays, while the cytokines were analyzed by ELISA. The EO treatments did not affect the viability; on the contrary, the PBMC proliferation increased in presence of all the EOs tested, according to the different percentages and mitogens used. The IL-10 secretion was higher in both the REO and the LEO tested at 3% than in the positive control; furthermore, the IL-8 level was influenced differently by the various EOs. The present data demonstrate that EOs may modulate the immune response activated by inflammation.

Role of hazelnut skin supplementation on plasma antioxidant status and cytokine profile in growing lambs.

In this study, the effect of hazelnut skin dietary supplementation on antioxidant status and cytokine profile was evaluated in growing lambs. A total of 22 male lambs at the age of 2 months, balanced for their initial live weight (15.33 ± SD 1.79 kg), were selected and allocated into two experimental groups: the control group (CON) receiving a maize-barley-based concentrated diet, and the hazelnut group (HS) receiving supplementation with hazelnut skin (150 g/kg on the dry matter) as a maize substitute for the concentrate diet. The experiment lasted for 56 days. Peripheral blood was collected at 7, 35, and 56 days of the experiment. The free radical scavenging activity using 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging assay, the total antioxidant capacity assay (TAC), the reactive oxygen species (ROS), and reactive nitrogen species (RNS) were determined in plasma. The secretion of IL-1ß, IL-6, and IL-10 cytokines was also determined by ELISA. The DPPH was affected by the interaction between feeding strategy and time of sampling (p = 0.039) with a higher level of DPPH at 7 days in the HS group than the CON group. The time of sampling affected the levels of plasma TAC (p = 0.016), while the ROS/RNS levels showed a higher value in the HS group (p < 0.001), on average. The antioxidant/oxidant index, which combines the TAC and the ROS/RNS levels, was not affected by the inclusion of hazelnut skin in the diet (p = 0.394). The cytokine profile showed a lower IL-6 secretion at both 35 and 56 days than at 7 days, on average. Furthermore, the feeding treatment affected the IL-1ß level, showing a lower level in the HS group than in the CON group on average. Lambs from the HS group had higher IL-10 plasma levels than the CON group at 7 days of the experiment. The present data highlight an antioxidant effect and a modulatory role in the cytokine profile of HS supplementation in growing lambs.

First insight into extracellular vesicle-miRNA characterization in a sheep in vitro model of inflammation.

Extracellular vesicles (EVs) and their microRNA (miRNA) cargoes have garnered attention in the veterinary field for their regulatory role in various biological processes. This study aimed to (i) evaluate two techniques of EV isolation from sheep peripheral blood mononuclear cell (PBMC) supernatants using the ultracentrifugation (UC) and reagent (REA) methods and (ii) characterize the EV-miRNA profiles after an in vitro inflammatory environment mediated by lipopolysaccharides (LPS). Sheep peripheral blood was collected, and PBMCs were separated using a density gradient reagent. Subsequently, PBMCs were cultured at 37°C for 24 h (5% CO2), and the supernatants were collected to perform the EV isolation. The presence of CD81+ extracellular vesicle marker was determined, and the purity of isolated EVs was calculated as a ratio between the number of isolated EVs and the protein concentration. Moreover, the morphological characterization revealed mainly round-shaped structures with average sizes of 211 nm for EVs isolated by the UC method and 99 nm for EVs isolated by the REA method. Illumina NextSeq sequencing in a single-end mode was used to characterize the miRNA profile, and the differentially expressed (DE) miRNAs were analyzed using a combination of bioinformatics tools. The results revealed that the REA method is reliable for EV isolation from sheep supernatants. It was considered an improvement of the recovery rate and purity of EVs with the enhancement of the number and the expression levels of characterized miRNAs. The EVs isolated by the UC method after an LPS challenge showed 11 DE miRNAs, among which eight miRNAs were upregulated and three were downregulated. On the other hand, the REA method revealed an EV cargo in which eight DE miRNAs were upregulated and 21 DE miRNAs were downregulated. The master miRNA regulators of the biological process were identified by performing the MIRNA-mRNA network analysis, showing that, among the higher representative miRNAs based on the centrality and betweenness, the miR-26a-5p could have a crucial role in the resolution of inflammation. Moreover, the identification of the let-7 miRNA family in all the EVs showed potential targeted genes that regulate the inflammation and immune responses.

Melatonin Administration to Pregnant Ewes for Coccidiosis Control in Their Offspring.

In livestock research, there has been a growing interest in the impact of melatonin on both health and disease conditions. The hypothesis of the present study was that melatonin treatment prenatally could support the immune competence and growth of experimentally infected lambs. This is the first study that aimed to investigate the impact of melatonin administration throughout pregnancy on immunity and oocyst excretion of pre-partum ewes and their offspring after experimental infection with Eimeria species. Thirty pregnant ewes were allocated into five equal groups, ΚΜ, ΚC, CM, CC, and NC, and gave birth to 47 lambs. Ewes of the KM and KC groups were orally challenged with a cocktail of Eimeria-sporulated oocysts (mainly consisting of Eimeria ovinoidalis), on day 120 of pregnancy, as well as all the lambs at the age of 5-9 days apart from those born from the NC group (environmental control). Fecal samples were collected from all ewes before infection and at parturition and from all lambs 14 times (S0-S13), before infection and during the following 8 weeks, for counting oocysts per gram of feces (OPG). Immunoglobulin (IgG) and cytokine (IL-1ß, IL-6, IL-10, IFN-γ) levels were determined in ewes' plasma collected before infection and at parturition, in lambs' plasma at 24 and 72 h after their birth, and in colostrum samples at parturition and 72 h later. Body weight of lambs was recorded five times from birth until the age of 60 days. Accordingly, the leucogram was evaluated in blood samples collected six times within the same period. On average, IgG concentration was higher (p < 0.05) in the blood of KM-ewes compared to KC and CC groups and in colostrum of KM-ewes compared to other groups (p < 0.001). KM-lambs had greater IgG titer and IFN-γ level than the other groups (p < 0.05). The IL-10/ IFN-γ ratio in KM-ewes was lower than the CC group (p = 0.06). Overall, the growth rate of lambs did not differ among groups (p > 0.05). Total oocysts' excretion in KM- and CM-lambs was reduced by 94.9% (p = 0.05) and 92.6% (p = 0.025), respectively, compared to KC-lambs, following the 3-week period after challenge, when E. ovinoidalis predominated in all groups. The dominant type of leucocytes was monocytes in all experimentally infected lambs, but not in NC-lambs, while overall lymphocytes were lower in KC-lambs than in NC-lambs (p < 0.05). Considering that almost all young indoor-reared lambs are exposed to coccidia species during their early life, melatonin treatment prenatally could suggest an alternative management tool in alleviating infection pressure.

Effects of management strategies on animal welfare and productivity under heat stress: A synthesis.

Climate change includes different dramatic events, and among them, heat stress exposition is the strongest phenomenon affecting the livestock sector. The effects of heat stress events on animal welfare are complex and the economic impacts for the livestock sector are relevant. Management measures may contribute to improve the resilience to heat stress, but the extent to which they impact on livestock performances and management strategies depend on the magnitude of the stress conditions. Through a pioneering synthesis of existing knowledge from experiments conducted in controlled conditions, we show that management strategies, both adaptation and mitigation measures, halved the negative impacts on the ruminants' performances and welfare induced by heat stress, but the efficacy is low in extreme conditions, which in turn are more and more frequent. These novel findings emphasize the need to deepen research on more effective adaptation and mitigation measures.

Proteomics in bovine semitendinosus muscle to assess emerging strategies based on papain injection and ultrasounds on meat tenderization process.

The impact of papain and/or ultrasound treatments on tenderization of semitendinosus muscle through a proteomic approach was studied. Sixteen bovine muscles were submitted to the following treatments: aging at 3 °C (Control), papain injection (PI), ultrasound (US), PI followed by US (PIUS) and US followed by PI (USPI). pH, myofibrillar fragmentation indices (MFI), soluble collagen, texture profile and changes of myofibrillar proteins were investigated after 2, 24, 48 and 96 h of storage. The highest MFI and soluble collagen content were found in PI, PIUS and USPI samples while control samples showed the lowest values. PI samples showed the lowest WBSF and hardness values until 48 h of storage while at 96 h meat from USPI treatment showed WBSF value comparable to PI treatment. The lowest values of cohesiveness, gumminess and chewiness were found in PI samples during all storage times. Proteomic analysis revealed a different quantity and expression of proteins among tenderization treatments. US treatment did not exhibit a significant ability to degrade muscle proteins, while, all treatments containing papain, showed a greater ability to hydrolyse and degrade myofibrillar proteins. PI promoted intense proteolysis leading to an early tenderization process; on the contrary, in PIUS and USPI treatments the sequence of treatments was relevant on meat tenderization. Particularly, USPI treatment, after 96 h, reached the same improvement in tenderness of enzymatic treatment but with slower hydrolysing rate; this could be determinant to preserve textural structure.

Patterns of Reproductive Management in Sheep and Goat Farms in Greece.

This paper reports findings regarding patterns of reproductive management in 325 sheep flocks and 119 goat herds throughout Greece. The objectives were to describe the patterns of reproductive management in sheep and goat farms, to highlight factors that were associated with these management patterns and the clinical application of the various reproductive management approaches in the farms and to establish potential associations with production outcomes. The median months of the start of the mating period for adult sheep and goats were May and June, respectively and for ewe lambs and replacement goats these were August and September, respectively. The median duration of the mating period was 2 months for adults and 1 month for replacements. The median ratio of female to male animals was 22:1 and 25:1 in sheep and goat farms, respectively. Control of reproduction was applied in 33.2% and 16.8% of sheep and goat farms, respectively, mostly by intravaginal progestagen sponges and less often by melatonin implants; multivariable analysis indicated five (availability of milking parlour, number of ewes in a farm, number of daily milkings, daily period spent by farmers at the farm, farming tradition in the family) and two (number of daily milkings, availability of milking parlour) predictors in sheep and goat farms, respectively. Pregnancy diagnosis ultrasonographically was performed in 36.6% and 16.8% of sheep and goat farms, respectively; multivariable analysis indicated as significant three (management system applied in farms, age of farmer, farming tradition in the family) and two (management system applied in farms, availability of milking parlour) predictors in sheep and goat farms, respectively. The control of reproduction and pregnancy diagnosis were associated with a higher number of vaccinations during pregnancy: 2.6 and 2.7, respectively, versus 2.2 and 2.1 in farms where they were not performed. The average per farm number of lambs/kids born per female animal was 1.33 ± 0.01 and 1.30 ± 0.02 for sheep and goat farms, respectively; five (control of reproduction, location of the farm, presence of working staff, collaboration with veterinary practice, breed of ewes) and two (control of reproduction, breed of female goats) predictors were identified in sheep and goat farms, respectively, for high numbers of lambs/kids. Lambs/kids were taken away from their dams at the age of 50 and 65 days, respectively. The average culling age (females, males) was 5.9 and 4.4 years and 6.9 and 4.9 years for sheep and goats, respectively. Farmers sourced animals to be used as replacements for those removed from the farm, mostly from the animals in their own farms, considering criteria related to milk production. Finally, the application of the control of reproduction was associated with increased milk production and the number of newborns, whilst an inverse correlation between the culling age of animals and milk production was seen.

Zoonotic Problems Reported by Sheep and Goat Farmers and Factors Potentially Contributing to the Occurrence of Brucellosis among Them.

The study aimed to investigate the occurrence of zoonotic problems reported by dairy small ruminant farmers in Greece and to study potential associations with socio-demographic characteristics of the farmers and management practices applied in the farms. A countrywide investigation was performed in 325 sheep and 119 goat farms in the 13 administrative regions of Greece. The selected farms were visited and interviews were conducted with respective farmers. The occurrence of zoonotic problems in the farmers was recorded. A total biosecurity score (0−6) was devised, based on biosecurity practices followed in farms. Sixty-seven farmers (15.10%, 95% confidence intervals (CI): 12.1−18.7%) reported experiencing a zoonotic problem. Most of the farmers (n = 57) (85.1%, 95% CI: 74.76−91.7%, of those with a zoonotic problem) (12.8%, 95% CI: 10.0−16.3%, of all) reported that the zoonotic problem had been brucellosis. Odds ratio for the occurrence of brucellosis in goat farmers was 1.879 (95% CI: 1.051−3.359) compared to the occurrence of the infection in sheep farmers (p = 0.033). For the outcome 'occurrence of brucellosis' in sheep farmers, the application of hand-milking, the availability of a separate lambing area and the presence of cats in the farm emerged as significant (p < 0.01); for the same outcome in goat farmers, only the availability of a separate kidding area emerged as significant (p = 0.001). The mean biosecurity score in farms in the continental area of the country was significantly higher than in the islands: 3.45 ± 0.05 versus 2.76 ± 0.28, respectively (p = 0.006), whilst there was also a significantly higher score in farms, where the farmer reported occurrence of brucellosis: 3.68 ± 0.15 versus 3.34 ± 0.06 in farms, where the farmer did not report such an incident (p = 0.042). In farms, where the above predictors prevail, farmers should be warned of an increased potential risk for human infection and biosecurity measures should be implemented and tightened.

Isolation of Listeria ivanovii from Bulk-Tank Milk of Sheep and Goat Farms-From Clinical Work to Bioinformatics Studies: Prevalence, Association with Milk Quality, Antibiotic Susceptibility, Predictors, Whole Genome Sequence and Phylogenetic Relationships.

A cross-sectional study was performed in 325 sheep and 119 goat dairy farms in Greece. Samples of bulk-tank milk were examined by standard microbiological techniques for Listeria spp. Listeria monocytogenes was isolated from one (0.3%) and Listeria ivanovii from three (0.9%) sheep farms. No associations between the isolation of L. monocytogenes or L. ivanovii and milk quality were found. No resistance to antibiotics was identified. Three variables emerged as significant predictors of isolation of the organism: the presence of pigs, low average relative humidity and a high number of ewes on the farm. The three L. ivanovii isolates were assessed in silico for identification of plasmids, prophages, antibiotic resistance genes, virulence factors, CRISPRs and CAS genes. Phylogenetic analysis using the core genome revealed that the three strains belonged to the L. ivanovii subsp. ivanovii branch and were especially close to the PAM 55 strain. All strains of the branch appeared to be very similar, with the distance between them being small.

Cytokine Pattern of Peripheral Blood Mononuclear Cells Isolated from Children Affected by Generalized Epilepsy Treated with Different Protein Fractions of Meat Sources.

The objective of the present study was the evaluation of cytokine patterns in terms of TNF-α, IL-10, IL-6, and IL-1ß secretion in peripheral blood mononuclear cell (PBMC) supernatants isolated from blood of children affected by generalized epilepsy and treated in vitro with myofibrillar, sarcoplasmic, and total protein fractions of meat and fish sources. Children with generalized epilepsy (EC group, n = 16) and children without any clinical signs of disease, representing a control group (CC group n = 16), were recruited at the Complex Structure of Neuropsychiatry Childhood-Adolescence of Policlinico Riuniti (Foggia, Italy). Myofibrillar (MYO), sarcoplasmic (SA), and total (TOT) protein fractions were obtained from longissimus thoracis muscle of beef (BF) and lamb (LA); from pectoralis muscle of chicken (CH); and from dorsal white muscle of sole (Solea solea, SO), European hake (Merluccius merluccius, EH), and sea bass fish (Dicentrarchus labrax, SB), respectively. PBMCs were isolated from peripheral blood of EC and CC groups, and an in vitro stimulation in the presence of 100 µg/mL for each protein fraction from different meat sources was performed. Data were classified according to three different levels of cytokines produced from the EC group relative to the CC group. TNF-α, IL-10, and IL-6 levels were not affected by different meat fractions and meat sources; on the contrary, IL-1ß levels were found to be significantly affected by the tested proteins fractions, as well as different meat sources, in high-level cytokine group. On average, the protein fractions obtained from LB, BF, and CH meat sources showed a higher level of IL-1ß than the protein fractions obtained from EH and SB fish samples. When all cytokine classes were analyzed, on average, a significant effect was observed for IL-10, IL-1ß, and TNF-α. Data obtained in the present study evidence that the nutritional strategy based on protein from fish and meat sources may modulate the immunological cytokine pattern of infants with generalized epilepsy.

High Milk Somatic Cell Counts and Increased Teladorsagia Burdens Overshadow Non-Infection-Related Factors as Predictors of Fat and Protein Content of Bulk-Tank Raw Milk in Sheep and Goat Farms.

This paper presents the results of an extensive countrywide investigation performed in 325 dairy sheep flocks and 119 goat herds throughout Greece. The objectives of the study were (a) to investigate fat and protein content in the bulk-tank raw milk of small ruminant farms in Greece and (b) to identify factors potentially influencing that content and factors that can contribute to increased content. The mean fat/protein contents in bulk-tank raw milk of sheep and goats were 6.16 ± 0.05%/4.43 ± 0.01% and 4.77 ± 0.44%/3.23 ± 0.30%, respectively. Significant differences were seen in protein content between farms in the various parts of the country. For sheep, multivariable analyses revealed breed and age of lamb removal from dams as significant for fat content, and somatic cell counts, management system in the farm, administration of anthelmintic treatment during the last stage of pregnancy, and farmer education as significant for protein content. For goats, significant factors were month into lactation period, age of kid removal from dams, and breed for fat content, and somatic cell counts, month into lactation, grazing duration, and % Teladorsagia larvae in faecal samples for protein content. For concurrently high fat and protein content, in multivariable analyses, the following emerged as significant factors: somatic cell counts in milk, numbers of parasite eggs in faeces, and veterinary collaboration (sheep), and month into lactation and somatic cell counts in milk (goats). The results indicate that high somatic cell counts in milk (reflecting the presence of mastitis) and gastrointestinal parasitic infections (mainly Teladorsagia infection) appear to exert a more significant influence on fat and protein content of milk, in comparison to non-infection-related factors.

Association of Melatonin Administration in Pregnant Ewes with Growth, Redox Status and Immunity of Their Offspring.

In this study, the effects of melatonin treatment on growth, redox status and immunity in prenatally stressed newborn lambs were evaluated. Thirty-seven newborn lambs were allocated into two groups (melatonin-MEL and control-CON), based on whether their mothers were treated with melatonin implants or not, respectively. All pregnant ewes were exposed to heat stress. The body weight of lambs was recorded at birth (L0), and then on days 15 (L15) and 40 (L40). Redox biomarkers [total antioxidant capacity (TAC), glutathione (GSH), thiobarbituric acid reactive substances (TBARS)] were assayed in blood samples collected from lambs on days L0, L1, L2, L5, L10 and L40. Chemical analysis and antioxidant capacity were evaluated in colostrum and milk samples collected at the same time points with blood samples. Cytokines (IL-1ß, IL-6, IL-10, IFN-γ) and immunoglobulin (IgG) were assayed in blood and colostrum samples collected from ewes on days L0 and L1, and in lambs' blood on days L0, L1 and L2. The results revealed that body weight gain of newborn lambs did not differ between the two groups (p > 0.05). Better redox status was found in MEL lambs until L2, as well as higher antioxidant capacity in the colostrum of MEL ewes compared to CON ones on day L0 (p < 0.05). In MEL ewes' colostrum, higher protein content was measured on day L0 and higher fat content on L1 compared to CON group (p < 0.05). The highest level of IL-6 was found in MEL ewes on L1, with a concomitant increase of IL-10 level in MEL lambs in comparison to CON lambs on L2. Moreover, CON colostrum resulted in a higher level of IL-10 within time, coupled with an increased level of IgG found in lambs' plasma on L2 (p = 0.04). This study indicated that melatonin could be administered as antioxidant and immune-modulatory regime in prenatally stressed offspring in order to cope with the crucial first days of their life. This effect of melatonin was also amplified by crosstalk between IL-6, IL-10 and IgG production, resulting in an improved quality of produced milk.

Lamb Meat Quality and Carcass Evaluation of Five Autochthonous Sheep Breeds: Towards Biodiversity Protection.

In the present study, the evaluation of the carcasses and meat quality, in terms of chemical composition and fatty acid profile, of lambs from five autochthonous sheep breeds (Altamurana, Bagnolese, Gentile di Puglia, Laticauda, and Leccese) reared in continental Southern Italy, were studied. All the carcasses were evaluated according to the EU Mediterranean classification system for carcasses weighing less than 13 kg. Meat chemical composition and fatty acids profile were assessed on both loin and leg commercial cuts. Fatty acid composition of loin resulted in differences among breeds, displaying lower values of saturated fatty acid in Altamurana, Bagnolese, and Leccese breeds and the highest content of polyunsaturated fatty acid in the Altamurana breed. Principal component analysis grouped lamb according to fatty acid content and to conjugated linoleic acid (CLA), omega n-3 and n-6 fatty acids; thus, Altamurana, Bagnolese, and Leccese breeds are characterized by the highest values of CLA content. Our data demonstrated that lamb meat from autochthonous breed has good carcass quality and the content of CLA, n-3, and n-6 was valuable for human consumption; therefore, the valorisation of local meat quality can help to avoid the extinction of the autochthonous breed offering to the market and consumer's high nutritive products.

Prevalence, Patterns, Association with Biofilm Formation, Effects on Milk Quality and Risk Factors for Antibiotic Resistance of Staphylococci from Bulk-Tank Milk of Goat Herds.

The objectives of this work were to study the prevalence and the patterns of antibiotic resistance of staphylococcal isolates from bulk-tank milk of goat herds across Greece, to assess possible associations of the presence of antibiotic resistance with the quality of milk in these herds and to evaluate herd-related factors potentially associated with the presence of antibiotic resistance among these staphylococcal isolates. A cross-sectional study was performed on 119 goat herds in Greece. Bulk-tank milk samples were collected for bacteriological examination; staphylococcal isolates were evaluated for resistance to 20 antibiotics. Oxacillin-resistant, resistant to at least one antibiotic, and multi-resistant staphylococcal isolates were recovered from 5.0%, 30.3%, and 16.0% of herds, respectively. Of 80 isolates, 7.5% were resistant to oxacillin, 50.0% were resistant to at least one antibiotic and 27.5% were multi-resistant. Resistance was seen more frequently among coagulase-negative staphylococci (59.3%) than among Staphylococcus aureus (23.8%). Resistance was more frequent against penicillin and ampicillin (41.3% of isolates) and fosfomycin (27.5%). No association was found with biofilm formation by staphylococci. For recovery of oxacillin-resistant isolates, the presence of working staff in the herds emerged as a significant factor; respective factors for the isolation of staphylococci resistant to at least one antibiotic were part-time farming and high (>10) number of systemic disinfections in the farm annually. The same three factors concurrently were also identified to be significant for the recovery of multi-resistant isolates.

Antibiotic Resistance of Staphylococci from Bulk-Tank Milk of Sheep Flocks: Prevalence, Patterns, Association with Biofilm Formation, Effects on Milk Quality, and Risk Factors.

The objectives of this work were to study prevalence and characteristics of resistance to antibiotics of staphylococcal isolates from the bulk-tank milk of sheep flocks across Greece, to assess possible associations of the presence of antibiotic resistance with the quality of milk in these flocks and to evaluate flock-related factors potentially associated with antibiotic resistance among these isolates. A cross-sectional study was performed in 325 sheep flocks in Greece. Bulk-tank milk samples were collected for bacteriological examination; staphylococcal isolates were evaluated for resistance to 20 antibiotics. Oxacillin-resistant staphylococcal isolates, isolates resistant to any antibiotic, and multi-resistant isolates were recovered from 8.0%, 30.5%, and 12.0% of flocks, respectively. Of 232 isolates, 11.6% were resistant to oxacillin, 46.1% were resistant to at least one antibiotic, and 16.4% were multi-resistant. Resistance was seen more frequently among coagulase-negative (50.6%) than among Staphylococcus aureus (31.5%) isolates. Resistance was more frequent against penicillin and ampicillin (34.1% of isolates), clindamycin (17.7%), and fosfomycin (14.2%). An association was found between biofilm formation by staphylococci and resistance to fosfomycin. For recovery of oxacillin-resistant isolates, the lack of experience by farmers emerged as a significant factor; respective factors for the isolation of staphylococci resistant to any antibiotic or multi-resistant isolates were the early stage of the lactation period (0th-1st month) and the intensive management system applied in the flocks, respectively.

Extensive countrywide field investigation of somatic cell counts and total bacterial counts in bulk tank raw milk in goat herds in Greece.

Dairy goat farming is an important sector of the agricultural industry in Greece, with an annual total milk production exceeding 450 000 l and accounting for over 25% of all goat milk produced in the European Union; this milk is used mainly for cheese production. Despite the importance of goat milk for the agricultural sector in Greece, no systematic countrywide investigations in the bulk-tank milk of goats in Greece have been reported. Objectives were to investigate somatic cell counts (SCC) and total bacterial counts (TBC) in raw bulk-tank milk of goat herds in Greece, study factors influencing SCC and TBC therein and evaluate their possible associations with milk content. Throughout Greece, 119 dairy goat herds were visited for milk sampling for somatic cell counting, microbiological examination and composition measurement. Geometric mean SCC and TBC were 0.838 × 106 cells ml-1 and 581 × 103 cfu ml-1, respectively. Multivariable analyses revealed annual frequency of check-ups of milking system and total milk quantity per goat (among 53 variables) to be significant for increased SCC; no factor emerged (among 58 variables) to be significant for increased TBC. Negative correlation of SCC with total protein was found; mean total protein content in the bulk-tank milk in herds with SCC >0.75 × 106 cells ml-1 was 5.1% lower and in herds with SCC >1.5 × 106 cells ml-1, it was 7.8% lower.

Assessed versus Perceived Risks: Innovative Communications in Agri-Food Supply Chains.

Food preparations, especially those based on animal products, are often accused of being responsible for the increase in food-borne infections, contributing to increased pressure on healthcare systems. The risk assessment in agri-food supply chains is of utmost importance for the food industry and for policymakers. A wrong perception of risks may alter the functioning of supply chains; thus, efforts should be devoted to communicating risks in an efficient way. We adopt a multidisciplinary approach to investigate how consumers perceive different food risks. Our analysis shows that planning effective communication strategies is very much important for efficiently informing consumers on food risks. We also comment on potential innovative ways to better organise the supply chains.

NETosis of Peripheral Neutrophils Isolated From Dairy Cows Fed Olive Pomace.

Neutrophils represent primary mobile phagocytes recruited to the site of infection, and their functions are essential to enhance animals' health performance. Neutrophils have an essential role in innate immunity and are able to kill the pathogens via the synthesis of neutrophil extracellular traps (NETs). The objective of the present work was the study of the in vitro NETosis of peripheral neutrophils isolated from dairy cows supplemented with olive pomace. Dairy cows (n = 16) balanced for parity (3.67 ± 1.5 for CON, 3.67 ± 1.9 for OP), milk yield (24.3 ± 4.5 kg d-1for CON and 24.9 ± 1.7 kg d-1 for OP), the number of days in milk (109 ± 83.5 for CON and 196 ± 51 for OP), and body weight (647 ± 44.3 kg for CON and 675 ± 70.7 kg for OP) were divided into two experimental groups fed with a control diet (CON) and supplemented with 6% of olive pomace (OP). Peripheral blood neutrophils were isolated and stimulated in vitro with phorbol-myristate-acetate (PMA) as a marker for activation and reactivity of the neutrophils. After isolation, both the viability and CD11b expression were analyzed by flow cytometry. Both NETosis by neutrophil elastase-DNA complex system and myeloperoxidase (MPO) activity were evaluated by ELISA. The specific antibodies against MPO and citrullination of Histone-H1 were used for investigating NETosis by immunofluorescence microscopy. The neutrophil elastase-DNA complexes produced during NETosis and MPO activity of neutrophil extracts were affected by OP supplementation. Furthermore, results from immunofluorescence analysis of NETosis depicted a similar result found by ELISA showing a higher expression of MPO and citrullination of Histone-H1 in OP than the CON neutrophils. In addition, all data showed that the OP diet resulted in a better response of neutrophils to PMA stimulation than the CON diet, which did not support the neutrophils' responses to PMA stimulation. Our results demonstrated that OP supplementation can enhance the neutrophil function in dairy cows leading to udder defense and inflammation response especially when an immunosuppression state can occur.

Extensive Countrywide Field Investigation of Somatic Cell Counts and Total Bacterial Counts in Bulk-Tank Raw Milk in Sheep Flocks in Greece.

Objectives were to investigate somatic cell counts (SCC) and total bacterial counts (TBC) in the raw bulk-tank milk of sheep flocks in Greece, to study factors potentially influencing increased SCC and TBC in the bulk-tank milk of sheep and to evaluate possible associations of SCC and TBC with milk content. Throughout Greece, 325 dairy sheep flocks were visited for collection of milk sampling for somatic cell counting, microbiological examination and composition measurement. Geometric mean SCC were 0.488 × 106 cells mL-1; geometric mean TBC were 398 × 103 cfu mL-1; 228 staphylococcal isolates were recovered form 206 flocks (63.4%). Multivariable analyses revealed annual incidence risk of clinical mastitis, age of the farmer and month into lactation period (among 53 variables) to be significant for SCC > 1.0 × 106 cells mL-1 and month into lactation period at sampling and availability of mechanical ventilators (among 58 variables) to be significant for TBC > 1500 × 103 cfu mL-1. Negative correlation of SCC with fat, total protein and lactose and positive correlation of SCC with added water were found. With SCC > 1.0 × 106 cells mL-1, significant reduction of protein content (2%) was observed, whilst in flocks with SCC > 1.5 × 106 cells mL-1, significantly lower annual milk production per ewe (42.9%) was recorded.