Innovative Use of Olive, Winery and Cheese Waste By-Products as Novel Ingredients in Weaned Pigs Nutrition

New sustainable sources of feeds, which can enhance the health and welfare of farm animals, lower feeding costs, and lead to safer products, are of high priority in modern animal production systems. In the present study, a novel silage created from Greek olive, winery, and feta cheese waste by-products, was evaluated as a feed ingredient at different inclusion rates (0%, 5% or 10%) in 34-day-old weaned pigs. The potential beneficial effects on performance, health and intestinal digesta microflora balance of the pigs were evaluated. Additionally, chemical, microbiological and quality analysis of the meat was carried out. Results showed no detrimental effects (p > 0.05) on the pigs’ performance and no significant changes (p > 0.05) in meat pH, color and chemical analysis. Ileum and cecum microflora populations (total anaerobes, Lactobacillaceae) were positively affected (p ≤ 0.05) by the dietary usage of the silage. The microbial populations (Clostridium spp.) of belly meat cuts were positively modified (p ≤ 0.01). The concentration of total phenols in the meat cuts were increased (p ≤ 0.05) and their resistance to oxidation was improved (p ≤ 0.05). In addition, the fatty acid profile of the meat lipids (polyunsaturated and n-3 fatty acids) was positively modified (p ≤ 0.001)

Electrospun Scaffolds as Antimicrobial Herbal Extract Delivery Vehicles for Wound Healing

Herbal extracts have been used in traditional remedies since the earliest myths. They have excellent antimicrobial, anti-inflammatory, and antioxidant activities owing to various bioactive components in their structure. However, due to their inability to reach a target and low biostability, their use with a delivery vehicle has come into prominence. For this purpose, electrospun nanofibrous scaffolds have been widely preferred for the delivery and release of antimicrobial herbal extracts due to the flexibility and operational versatility of the electrospinning technique. Herein, we briefly reviewed the electrospun nanofibrous scaffolds as delivery systems for herbal extracts with a particular focus on the preclinical studies for wound-healing applications that have been published in the last five years. We also discussed the indirect effects of herbal extracts on wound healing by altering the characteristics of electrospun mats

Safety Issues Regarding the Detection of Antibiotics Residues, Microbial Indicators and Somatic Cell Counts in Ewes’ and Goats’ Milk Reared in Two Different Farming Systems

Milk samples of ewes’ and goats’ unprocessed milk were collected from milk tanks from various farms in Epirus, Greece and classified in two groups according to the type of farming. Analyses of the samples included microbial inhibition assays to detect the presence of antibiotic residues, isolation of Staphylococcus aureus and Escherichia coli strains as microbiological indicators for susceptibility to antimicrobial medicines, Somatic Cells and Coagulase Negative Staphylococci (CNS) counts. These findings were correlated with each other as well as to the stage of the lactation period and to the type of the farming practices. Monitoring of bulk tank milk for residues of antibiotics should be performed after heating of the milk, on a regular basis, and should include at least two different tests. The results point out that the type of farming affects the CNS counts as well as the prevalence of residues in the milk. Furthermore, the inverse correlation between CNS counts and prevalence of residues of antibiotics suggests a possible protective role of CNS. Resistance of the bacterial indicators to antibiotics was random and relatively rare, perhaps acquired in past due to misuse of antibiotics, turning the indicator microorganisms to reservoir of resistance

The Assessment of Antimicrobial and Anti-Biofilm Activity of Essential Oils against <i>Staphylococcus aureus</i> Strains

The increase in antimicrobial resistance and tolerance over the years has become a serious public health problem, leading to the inevitable development of alternative antimicrobial agents as substitutes for industrial pharmaceutical antibiotics targeting humans and animals under the concept of one health. Essential oils (EOs) extracted from aromatic and pharmaceutical plants incorporate several bioactive compounds (phytochemicals) that positively affect human and animal health. Herein, this work aimed to examine a standardized chemical composition and screen the antimicrobial and anti-biofilm activity of Thymus sibthorpii, Origanum vulgare, Salvia fruticosa, and Crithmum maritimum EOs against three different Staphylococcus aureus strains by gold-standard disc diffusion, broth microdilution, and microtiter plate biofilm assays. Therefore, the evaluation of the above-mentioned EOs were considered as substitutes for antibiotics to combat the ever-mounting antimicrobial resistance problem. The observed bacterial growth inhibition varied significantly depending on the type and concentration of the antimicrobials. Thymus sibthorpii was determined as the strongest antimicrobial, with 0.091 mg/mL minimum inhibitory concentration (MIC) and a 14–33 mm diameter inhibition zone at 5% (v/v) concentration. All tested EOs indicated almost 95% inhibition of biofilm formation at their half MIC, while gentamicin sulfate did not show sufficient anti-biofilm activity. None of the methicillin-resistant strains showed resistance to the EOs compared to methicillin-sensitive strains. Thymus sibthorpii and Origanum vulgare could be potential alternatives as antimicrobial agents to overcome the problem of microbial resistance. The tested EOs might be incorporated into antimicrobial products as safe and potent antimicrobial and anti-biofilm agents

Innovative Use of Olive, Winery and Cheese Waste By-Products as Functional Ingredients in Broiler Nutrition

The purpose of this study was to evaluate the dietary use of novel silage that was created by combining three agro-industrial wastes produced in bulk, i.e., olive mill wastewater, grape pomace, and deproteinized feta cheese whey, in the diets of broiler chickens. A total of 216 one-day-old male Ross-308 chicks were randomly allocated to three treatment groups with six replications (12 chicks per pen). Three isocaloric and isonitrogenous diets were formulated to include the examined silage at 0%, 5%, or 10%. Commercial breeding and management procedures were employed throughout the trial. At the end of the trial (day 35), tissue samples were collected for analysis. Feeding 10% silage resulted in increased (p &le; 0.001) final body weight (p &le; 0.001) and feed intake. Jejunum and cecum microflora, as well as breast and thigh meat microflora, were modified (p &le; 0.05) by the dietary inclusion. Thigh meat oxidative stability was improved (p &lt; 0.01) by the silage supplementation. In addition, breast and thigh meat fatty acid profiles were different, respectively, (p &lt; 0.05) in the supplemented treatments compared to the control. The examined silage was successfully tested in broiler diets with potential benefits for their performance and meat quality

Identification by MALDI-TOF MS and Antibiotic Resistance of <i>Riemerella anatipestifer</i>, Isolated from a Clinical Case in Commercial Broiler Chickens

The Gram-negative bacterium Riemerella anatipestifer (RA) is known to cause clinical disease with severe economic impacts primarily in ducks and less frequently in geese and turkeys. RA was isolated and identified in broiler chickens, from a clinical case in a commercial broiler farm located in the southwest mainland of Greece. The morbidity and the mortality in the broiler house were estimated at 10% and 5% respectively. The observed clinical signs appeared at the age of 30 to 42 days with respiratory distress (dyspnea), white fluid feces and stunting. Post-mortem examinations displayed serositis, pericarditis, perihepatitis and airsacculitis. Edematous swelling around the tibio-tarsal joints was observed in some birds. Tissue samples from lesions were streaked on selective media. Three bacterial isolates were identified by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS). Moreover, an antibiogram analysis was performed for the three RA strains, using a pattern of 16 common antibiotics to advocate the most effective drugs for a proper treatment. All the RA isolates were sensitive to ceftiofur, sulphamethoxazole–trimethoprim and amoxicillin, whereas all were resistant to gentamicin, tylosin, tetracyclin, colistin sulphate, spectinomycin, lincomycin and oxytetracycline

Overview of Ecology and Aspects of Antibiotic Resistance in <i>Campylobacter</i> spp. Isolated from Free-Grazing Chicken Tissues in Rural Households

Rural households all over the world rear backyard chicken mainly for their own consumption and, to a lesser extent, for barter trade. These chickens represent a staple dish with numerous culinary variations and a cheap source of protein. Although some Campylobacter species, and particularly Campylobacter jejuni and Campylobacter coli, have been associated with industrial poultry carcasses, studies concerning the ecology of this genus in rural households do not exist. To assess the prevalence of Campylobacter species in the tissues of backyard chickens, samples were collected from birds Gallus domesticus bred in households in the rural area of Epirus (Greece), and Campylobacter strains were isolated by quantitative methods at 37 °C and 42 °C. In total, 256 strains were identified, belonging to 17 Campylobacter species, with C. jejuni and C. coli being the most prevalent. From the four ecological parameters studied (size of the flock, presence of small ruminants in the same household, presence of other poultry species in the same household, and feeding leftovers of the household), the size of the flock and the presence of small ruminants and/or pigs in the same household mostly affected the distribution of these strains. To study the phenotypical resistance against 14 antibiotics, 215 strains were selected. The results showed a high prevalence of multidrug-resistance (MDR) strains extending to all classes of antibiotics. Further genome analysis revealed the presence of genes coding resistance (blaOxA-61, tet(O), tet(A) cmeA, cmeB, cmeC, and gyrA (Thr-86-Ile mutation)), with the efflux pump CmeABC being the most prevalent. All antimicrobial resistance-encoded genes co-circulated, except for blaOXA-61, which moved independently. The minimum inhibitory concentration (MIC) values of two out of three antibiotics (representing different classes) were reduced when the strains tested were exposed to carbonyl cyanide 3-chlorophenylhydrazone (CCCP), a known efflux pump inhibitor. The same result was obtained with the addition of CCCP to the MIC values of bile salts. These results lead to the conclusion that Campylobacter species are present in an impressive diversity in backyard chicken tissues and that they exert a significant resistance to antibiotics, raising a potential danger for public health

Antimicrobial and Methicillin Resistance Pattern of Potential Mastitis-Inducing <i>Staphylococcus aureus</i> and Coagulase-Negative Staphylococci Isolates from the Mammary Secretion of Dairy Goats

Staphylococcus spp. is an important mastitis-inducing zoonotic pathogen in goats and is associated with antimicrobial resistance (AMR). The objectives of this study were to determine the prevalence and composition of staphylococci in individual mammary secretion (MS) samples of clinically healthy goats and to evaluate the phenotypic AMR pattern and the presence of methicillin resistance in the Staphylococcus spp. strains. Staphylococcus spp. isolates (n = 101) from the MS samples (n = 220) were identified to species level using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry. The antimicrobial susceptibility testing included a disk diffusion assay and the determination of the minimum inhibitory concentrations (MIC) of resistant strains (n = 46). Presumptive methicillin-resistant strains (n = 9) were assessed for the presence of mecA, mecC and SCCmec/orfx genes. Staphylococcus spp. isolates were recovered from 45.9% of the MS samples, of which, 72.3% was identified as coagulase-negative staphylococci (CoNS), with the remaining being Staphylococcus aureus. CoNS and S. aureus were most commonly resistant to ampicillin (56.2% and 57.1%, respectively), penicillin (26.0% and 39.3%, respectively), amoxicillin (26 % and 25 %, respectively) and cephalexin (12.3% and 25%, respectively) in the disk diffusion method. CoNS exhibited a broader AMR pattern and a higher percentage of resistant strains than S. aureus in the disk diffusion and MIC methods. Of the nine oxacillin- and cefoxitin-resistant strains, three S. aureus and five CoNS strains carried the mecA gene and, thus, were identified as methicillin-resistant. The mecC gene was not found in any of the studied strains. The presence of AMR and methicillin resistance in caprine S. aureus and CoNS poses a concern for animal and public health

Effects of Dietary Omega-3 Enrichment on the Chemical Composition and the Pathogenic Microbiota of Ovine Milk

The demand for ovine milk and ovine dairy products is constantly increasing due to their exceptional sensorial characteristics and their health benefits for consumers. However, dairy fat content and composition are of particular concern for consumers as well as the medical community, as there are risk factors for coronary disease, diabetes mellitus, cancer, and other serious diseases. For this reason, attempts have been made to control/regulate the fat composition of ovine milk by modifying sheep dietary intake of polyunsaturated fatty acids. In this experimental trial, a group of sheep were fed for 30 days a diet enriched in flaxseeds and lupines, feed ingredients rich in omega-3 fatty acids, aiming to investigate the effects on fat composition and the microbiota of ovine milk. Chemical analysis of the collected milk showed that the omega-3 and omega-6 content was increased. On the opposite, the atherogenic and thrombogenic indexes decreased. Of importance was the semi-protective effect on the udder by the increased omega-3 dietary intake, as depicted by its impact on the biodiversity of the pathogenic microbiota. These findings suggest that ovine milk could be modified under specific conditions to be more appropriate for the consumption by people belonging to high-risk groups for various diseases

Effect of an Herbal Mixture of Oregano, Garlic, Sage and Rock Samphire Extracts in Combination with Tributyrin on Growth Performance, Intestinal Microbiota and Morphology, and Meat Quality in Broilers

The present study investigated the effects of two feed additives, the first containing an herbal mixture of oregano, garlic, sage, and rock samphire extracts and the second containing tributyrin (glyceryl tributyrate) when fed to broiler chickens. A total of 360 one-day-old chicks were randomly allocated to four treatments (6 replicate pens of 15 chicks). One treatment served as the unsupplemented control, whereas the feeds of the other three treatments were supplemented either with the herbal additive (3 g/kg), the tributyrin additive (1 g/kg), or both additives. The duration of the trial was 37 days. Data were collected on growth performance, intestinal microbiota and morphology, and some meat quality parameters. The combined supplementation improved (p &lt; 0.05) weigh gain, feed conversion ratio, and the European Efficiency Factor. In the cecum, the combined supplementation lowered (p &lt; 0.05) the microbial populations of aerobes, anaerobes, Escherichia coli, total Enterobacteriaceae, and Clostridium spp. compared to the other treatments. Fecal coccidial oocyst counts were also reduced (p &lt; 0.01) by the combined supplementation. The herbal mixture supplementation improved (p &lt; 0.05) breast and thigh meat resistance to oxidation. In conclusion, the combined dietary supplementation with the examined feed additives could be utilized to improve the performance and intestinal health of broiler chickens