Assessment of the feed additive consisting of Levilactobacillus brevis DSM 16680 for all animal species for the renewal of its authorisation (Microferm Ltd.).

Following a request from the European Commission, EFSA was asked to deliver a scientific opinion on the assessment of the application of renewal of Levilactobacillus brevis DSM 16680 as a technological feed additive (functional group: silage additives) for all animal species. The applicant has provided evidence that the additive currently on the market complies with the existing terms of the authorisation. The EFSA Panel on Additives and Products or Substances used in Animal Feed (FEEDAP) concluded that the additive remains safe for all animal species, consumers and the environment. Regarding user safety, the additive should be considered as an eye irritant and a skin and respiratory sensitiser. There is no need for assessing the efficacy of the additive in the context of the renewal of the authorisation.

Modification of the terms of authorisation regarding the additive consisting of liquid l-lysine base produced with Corynebacterium glutamicum NRRL B-67439 and NRRL B-67535 for all animal species (ADM specialty ingredients (Europe) B.V.).

Following a request from the European Commission, the EFSA Panel on Additives and Products or Substances used in Animal Feed (FEEDAP) was asked to deliver a scientific opinion on the safety and efficacy of liquid l-lysine base produced with a genetically modified strain of Corynebacterium glutamicum as a nutritional feed additive for all animal species. The l-lysine base liquid produced with C. glutamicum NRRL B-67535 and NRRL B-67439 is currently authorised as a nutritional additive for all animal species. The present application is aimed at modifying the current authorisation to include C. glutamicum NRRL B-68248 as a production strain. The new production strain qualifies for the qualified presumption of safety approach when used for production purposes. It was unambiguously identified as C. glutamicum and was shown not to harbour acquired antimicrobial resistance determinants for antibiotics of human and veterinary importance. All the introduced sequences or mutations were considered to be safe, and no viable cells or DNA of the NRRL B-68248 strain was detected in the final product. Therefore, the final product does not pose any safety concern associated with the production strain. l-Lysine base produced using C. glutamicum NRRL B-68248 does not represent a risk for the target species, the consumer or the environment. The additive was considered to be neither irritant to skin or the eyes, nor a dermal sensitiser. l-Lysine base produced with C. glutamicum NRRL B-68248 is considered to be an efficacious source of the essential amino acid l-lysine for non-ruminant animal species. For the supplemental l-lysine to be as efficacious in ruminants as in non-ruminant species, it would require protection against degradation in the rumen.

Assessment of the feed additive consisting of sodium propionate for all terrestrial animal species for the renewal of its authorisation (BASF SE).

Sodium propionate is authorised containing at least 98.5% of sodium propionate. The applicants requested for the renewal of the authorisation of sodium propionate when used as a feed additive for all terrestrial animal species. The applicant has provided evidence that the additive in the market complies with the conditions of the authorisation. The Panel on Additives and Products or Substances used in Animal Feed (FEEDAP Panel) confirms that the use of sodium propionate under the current authorised conditions of use is safe for the target species, the consumers and the environment. Considering the user safety, the additive is corrosive to skin, eyes and respiratory tract, but is not a skin sensitiser. There is no need for assessing the efficacy of the additive in the context of the renewal of the authorisation.

Assessment of the feed additive consisting of endo-1,4-beta-xylanase (produced with Trichoderma reesei MUCL 49755) and endo-1,3(4)-beta-glucanase (produced with T. reesei MUCL 49754) (AveMix® XG 10) for pigs for fattening, minor porcine species for fattening and turkeys for fattening for the renewal of its authorisation (AVEVE BV).

Following a request from the European Commission, EFSA was asked to deliver a scientific opinion on the assessment of the feed additive consisting of endo-1,4-beta-xylanase (produced with Trichoderma reesei MUCL 49755) and endo-1,3(4)-beta-glucanase (produced with T. reesei MUCL 49754) (AveMix® XG 10/AveMix® XG 10 L) for the renewal of its authorisation as zootechnical feed additive for pigs for fattening, minor porcine species for fattening and turkeys for fattening. The applicant declared a change in the carrier material used in AveMix® XG 10 from soybean meal to calcium carbonate + wheat flour or calcium carbonate + sepiolite. The applicant provided evidence that the additive Avemix® XG 10 with calcium carbonate + wheat flour and Avemix® XG 10 L comply with the conditions of the authorisation. The EFSA Panel on Additives and Products or Substances used in Animal Feed (FEEDAP) noted that no data were submitted to support compliance of the formulation of Avemix® XG 10 with calcium carbonate + sepiolite with the conditions of the authorisation. The FEEDAP Panel concluded that both forms of the additive remain safe for pigs for fattening, minor porcine species for fattening and turkeys for fattening, consumers and the environment. Regarding the safety for the user, Avemix® XG 10 formulated with calcium carbonate + sepiolite and Avemix® XG 10 L are not irritant to skin and eyes. No conclusions on the irritation potential of Avemix® XG 10 formulated with calcium carbonate + wheat flour could be drawn. The additive in all its formulations is considered a respiratory and skin sensitiser. There was no need for assessing the efficacy of the additive in the context of the renewal of the authorisation.

Larval performance of Zophobas morio (F.) (Coleoptera: Tenebrionidae) on various diets enriched with post-distillation residues and essential oils of aromatic and medicinal plants.

The increasing demands for resources driven by the global population necessitate exploring sustainable alternatives for affordable animal protein over the use of traditional protein sources. Insects, with their high protein content, offer a promising solution, especially when reared on agricultural post-distillation residues for enhanced sustainability and cost-effectiveness. We assessed the development of Zophobas morio (F.) (Coleoptera: Tenebrionidae) larvae on diets enriched with essential oils and post-distillation residues from Greek aromatic and medicinal plants. Two aromatic plant mixtures (A and B) were examined. Mixture A consisted of post-distillation residues, while Mixture B incorporated these residues along with essential oils. Insect rearing diets were enriched with different proportions (10, 20, and 30 %) of these mixtures, with wheat bran serving as the control. Enrichment positively influenced larval development without compromising survival. Larval weight remained unchanged with Mixture A, but improved with Mixture B. No adverse effects were detected in the case of the enriched diets, although higher concentrations of Mixture B prolonged development time.

Combined Dietary Supplementation of Tenebrio molitor Larvae and Chitosan in Growing Pigs: A Pilot Study.

Nowadays, the global animal industry faces considerable challenges in securing sufficient feed resources. Responding to consumer demands for reduced use of antibiotics in animal nutrition, better animal welfare status, and reduced impact on the environment, there is an increased urgency to develop innovative functional feeds with a reduced environmental footprint and the ability to improve meat quality and safety. In an effort to explore innovative feed ingredients for growing pig diets, the combined dietary supplementation of Tenebrio molitor larvae and chitosan was investigated. An experimental trial was performed with 48 weaned pigs (34 days of life; mixed sex) that were randomly assigned to four treatment groups (with six males and six females each): Group A (control), Group B (supplemented with T. molitor larvae at 10%), Group C (supplemented with chitosan at 0.05%), and Group D (supplemented with both ingredients at 10% and 0.05%, respectively). On the 42nd day of the experimental trial, samples of blood, feces, and carcass parts were taken for analysis. The results indicated that the insect larvae meal significantly improved (p < 0.05) overall performance, increased (p < 0.05) blood red blood cell content, increased meat phenolic content (p < 0.05), improved meat oxidative stability (p < 0.05), and affected meat fatty acid profile (p < 0.05). On the other hand, chitosan had no significant effect on overall performance (p > 0.05), but it significantly increased blood lymphocyte content (p < 0.05), affected the fecal microbiota (p < 0.05), improved meat oxidative stability (p < 0.05), increased meat phenolic content (p < 0.05), and affected meat fatty acid composition (p < 0.05) and (p < 0.05) meat color. Finally, the combined use of both T. molitor and chitosan significantly affected some important zootechnical parameters (p < 0.05), fecal microbial populations (p < 0.05), meat color (p < 0.05), and fatty acid profile (p < 0.05). Further investigation into the potential interaction between insect larvae meals and chitosan in pig diets is advised.

Breeding Substrate Containing Distillation Residues of Mediterranean Medicinal Aromatic Plants Modulates the Effects of Tenebrio molitor as Fishmeal Substitute on Blood Signal Transduction and WBC Activation of Gilthead Seabream (Sparus aurata).

This work assesses the dietary use of two insect meals of Tenebrio molitor (TM) larvae reared in conventional (TM-10) or MAP-enriched substrates (MAP-TM-10) as fish meal replacements (10%) in the diets of gilthead seabream (Sparus aurata). Fish (n = 4500; 207.19 ± 1.47 g) were divided into three groups with triplicates: control (fed conventional diet), TM-10, and MAP-TM-10 groups. The fish were reared in floating cages for 12 weeks and the dietary effects on white blood cell activation, heat shock proteins, MAPKs, and apoptosis of the fish were evaluated. The MAP-TM-10 group exhibited the highest eosinophilic induction. Phosphorylated levels of p38 MAPK, p44/42 MAPK, HSP70, and HSP90 increased in the TM-10 and MAP-TM-10 groups. In terms of apoptosis, Bax levels were lower in the TM groups compared to the control, and the MAP-TM-10 group showed even lower levels than the TM-10 group. Bcl-2 levels increased in the TM-10 group compared to the control, and further increased in the MAP-TM-10 group. The Bax/Bcl-2 ratio, an apoptosis indicator, decreased in the TM groups, with the MAP-TM-10 group showing a further decrease compared to TM-10. These findings suggest that insects' breeding substrate being enriched with MAPs modulated the effect of TM on cellular stress and apoptosis.

Use of an Innovative Silage of Agro-Industrial Waste By-Products in Pig Nutrition: A Pilot Study of Its Effects on the Pig Gastrointestinal Microbiota.

The aim of this study was to evaluate whether dietary supplementation with an innovative silage (IS) created using 60% olive mill waste, 20% grape pomace, and 20% deproteinised feta cheese waste solids can modulate the composition of the intestinal microbiota in weaned (Exp. 1) and finishing (Exp. 2) pigs. In Exp. 1 (40 day supplementation), forty-five crossbred weaned pigs were randomly assigned to the 0% (Control), 5%, or 10% IS groups (15 replicates/experimental diet). In Exp. 2 (60 day supplementation), eighteen finishing pigs from Exp. 1 were fed the control diet for 8 weeks before being re-assigned to their original experimental groups and fed with the 0% (Control), 5%, or 10% IS diets (six replicates/experimental diet). Performance parameters were recorded. Ileal and caecal digesta and mucosa were collected at the end of each experiment for microbiota analysis using 16S rRNA gene sequencing (five pigs/experimental diet for Exp. 1 and six pigs/experimental diet for Exp. 2). No significant effects on pig growth parameters were observed in both experiments. In Exp. 1, 5% IS supplementation increased the relative abundance of the Prevotellaceae family, Coprococcus genus, and Alloprevotella rava (OTU_48) and reduced the relative abundance of Lactobacillus genus in the caecum compared to the control and/or 10% IS diets (p < 0.05). In Exp. 2, 5% IS supplementation led to compositionally more diverse and different ileal and caecal microbiota compared to the control group (p < 0.05; p = 0.066 for ß-diversity in ileum). Supplementation with the 5% IS increased the relative abundance of Clostridium celatum/disporicum/saudiense (OTU_3) in the ileum and caecum and Bifidobacterium pseudolongum (OTU_17) in the caecum and reduced the relative abundance of Streptococcus gallolyticus/alactolyticus (OTU_2) in the caecum compared to the control diet (p < 0.05). Similar effects on C. celatum/disporicum/saudiense and S. gallolyticus/alactolyticus were observed with the 10% IS diet in the caecum (p < 0.05). IS has the potential to beneficially alter the composition of the gastrointestinal microbiota in pigs.

Utilization of Tenebrio molitor Larvae Reared with Different Substrates as Feed Ingredients in Growing Pigs.

The procurement of adequate feed resources is one of the most important challenges for the animal industry worldwide. While the need for feeds rich in protein is constantly increasing, their production cannot readily keep up. Consequently, to overcome this challenge in a sustainable way, it is necessary to identify and develop new feeding strategies and feed ingredients, such as insect meals. In the present study, Tenebrio molitor larvae that were reared on two different substrates (standard and enriched with medicinal aromatic plant material) were used as feed ingredients for growing pigs. A total of 36 weaned pigs (34 days old) were randomly allocated to three treatment groups and fed either the control diet (A) or diets supplemented at 10% with one of the two insect meals (B and C). At the end of the trial (42 days), blood, feces, and meat samples were collected for analysis. The insect meal supplementation did not affect (p > 0.05) overall performance but significantly modified (p < 0.001) the fecal microflora balance and the blood cholesterol (p < 0.001), while the rest of the blood parameters tested were not affected. Moreover, this dietary supplementation significantly affected some microbial populations (p < 0.001), improved the total phenolic content (p < 0.05), and the fatty acid profile (p < 0.001) of the meat cuts, but did not affect (p > 0.05) meat color or proximate composition. Further research is needed to evaluate the different types and levels of inclusion of insect meals in pig nutrition.

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).

Growth performance, welfare traits and meat characteristics of broilers fed diets partly replaced with whole Tenebrio molitor larvae.

The role of insects in animal nutrition has been reconsidered during recent years, paving the way for an increasing market for edible insects. Their protein and amino acid balance make them a promising source of protein for replacing high value proteins. Yellow mealworm, Tenebrio molitor L. (TM; Coleoptera: Tenebrionidae) larvae, have shown positive effects on broiler performance in several research studies and have a strong potential as a sustainable alternative protein source for monogastric animals. This study aimed to assess the effect of replacing various ratios of basal diets with T. molitor larvae on broiler performance as well as on several meat and welfare characteristics. For the study, 120 one-day-old male chicks (Ross 308) were randomly allocated in 3 treatments and 4 replications (10 birds per pen). Birds of the control group (basal diet) were fed with typical commercial maize and soymeal-based rations in mash form. The other 2 groups were treated with the same diet, after replacing 5% and 10% with dried TM larvae, respectively. On d 35 (end of trial), meat samples were collected and analysed. Body weight, feed intake, body weight gain and feed conversion ratio during the periods of 1 to 10 d (starter period), 11 to 24 d (grower period), 25 to 35 d (finisher period) and 1 to 35 d (total period) were assessed. Pododermatitis, diarrhoea, feather score and litter conditions were also assessed during the trial. The results indicated that TM larvae inclusion in the broilers' diet positively affected body weight gain values, as well as the carcass yield, the meat composition and various welfare traits. Additionally, the dietary treatments with TM larvae favourably affected meat composition and colour parameters, whereas there were also some positive effects on lipid and protein oxidation. Saturated fatty acids were decreased by the dietary supplementation whereas the polyunsaturated fatty acids to SFA ratio increased. In general, the study showed that whole TM larvae addition can provide a promising alternative to soybean meal in the diet of broilers, demonstrating a positive impact on growth, welfare and meat characteristics.

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.

In Vitro Antioxidant, Antimicrobial, Anticoccidial, and Anti-Inflammatory Study of Essential Oils of Oregano, Thyme, and Sage from Epirus, Greece.

Origanum vulgare subsp. hirtum, Thymus vulgaris, and Salvia fructicosa are aromatic plants commonly found in Mediterranean countries and are traditionally used in Greece as a remedy for humans, since they are well known as potent antibacterial, antioxidant, and anti-inflammatory agents. Essential oils (EOs) derived from plants cultivated in the mountainous region of Epirus, Greece, were investigated for their inhibitory activity against key microorganisms with relevance to avian health, while also assessing their antioxidant and anti-inflammatory activity. The total phenolic content (TPC) of the EOs was estimated according to the Folin−Ciocalteu method, while the antioxidant capacity was tested through the EOs' ability to scavenge free radicals by means of the DPPH, ABTS, and FRAP assays. Antibacterial and anti-inflammatory effects were examined by the agar disc diffusion method and the lipoxygenase (LOX) inhibition test, respectively. Furthermore, the EOs' ability to inhibit the invasion of sporozoites of Eimeria tenella (Wisconsin strain) along with any toxic effects were assayed in Madin−Darby bovine kidney (MDBK) cells. The antioxidant activity of the EOs was observed in descending order: oregano > thyme > sage. The antimicrobial effects of thyme and oregano were equivalent and higher than that of sage, while the anti-inflammatory effect of thyme was higher compared to both sage and oregano. The intracellular invasion of sporozoites was evaluated by the detection of E. tenella DNA by qPCR from cell monolayers harvested at 2 and 24 h post-infection. Parasite invasion was inhibited by the addition of oregano essential oil at the concentration of 100 µg/mL by 83% or 93% after 2 or 24 h, respectively, and was higher compared to the addition of thyme and sage, which had similar effects, but at a less intensive level. The cytotoxic assessment of all three essential oils revealed that they had no effect on MDBK cells compared to dimethyl sulfoxide (DMSO), used as the control substance. The supplementation of oregano, thyme, and sage essential oils had a potent antioxidant, anti-inflammatory, antimicrobial, and anticoccidial in vitro effect that is comparable to synthetic substances or approved drugs, justifying the need for further evaluation by in vivo studies in broilers reared in the absence of antimicrobial and anticoccidial drugs or synthetic antioxidant and/or anti-inflammatory compounds.

Antimicrobial and Methicillin Resistance Pattern of Potential Mastitis-Inducing Staphylococcus aureus 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.

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 ≤ 0.001) final body weight (p ≤ 0.001) and feed intake. Jejunum and cecum microflora, as well as breast and thigh meat microflora, were modified (p ≤ 0.05) by the dietary inclusion. Thigh meat oxidative stability was improved (p < 0.01) by the silage supplementation. In addition, breast and thigh meat fatty acid profiles were different, respectively, (p < 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.

Impact of an Omega-3-Enriched Sheep Diet on the Microbiota and Chemical Composition of Kefalograviera Cheese.

Kefalograviera is a well-known hard Greek cheese. The aim of this study was to determine how milk produced from ewes fed omega-3-enriched diets could influence the microbiota as well as the chemical composition of Kefalograviera cheese. At the start of the trial, 30 dairy ewes (Lesvos and Chios crossbreed) were selected and fed a conventional diet, based on alfalfa hay, straw and concentrate feed that contained soybean meal for a period of thirty days. Then, for a period of sixty days the same ewes were fed an omega-3-enriched concentrate feed with a lower level of soybean meal that contained 10% flaxseed and 10% lupins. Milk yield was collected individually on Days 30, 60 and 90 and used to produce three different batches of Kefalograviera cheeses, at the same cheese factory, by using a traditional recipe and identical preparation conditions (pasteurization of milk, salt, rennet and culture). Sample analysis was done after six months of Kefalograviera cheese ripening. MALDI-TOF-MS (matrix-assisted laser desorption/ionization time of flight mass spectrometry) identification was performed by contrasting the samples' mass spectra with the corresponding reference database. The correlation between the different Kefalograviera cheeses revealed the predominant species being Lactococcus lactis, Lactobacillus rhamnosus, Lactobacillus plantarum, Lactobacillus brevis, Lactobacillus paracasei, Enterococcus faecium and Enterococcus faecalis, with significant quantitative differences between the experimental groups and the controls. Pediococcus spp. was isolated only from the experimental groups' cheeses and Staphylococcus spp. only from the controls' cheese, suggesting-among other differences-a bacterial microbiota distinction between the groups. Moreover, increased levels of alpha-linolenic acid and total polyunsaturated omega-3 fatty acids were noted in the enriched Kefalograviera cheeses. These promising findings suggest that enriched Kefalograviera cheese could be manufactured via enriching the ewes' diets, with potential benefits for the consumers' health.

Reduction of Energy Intensity in Broiler Facilities: Methodology and Strategies.

Broiler facilities consume a lot of energy resulting in natural source depletion and greater greenhouse gas emissions. A way to assess the energy performance of a broiler facility is through an energy audit. In the present paper, an energy protocol for an energy audit is presented covering both phases of data collection and data elaboration. The operational rating phase is analytically and extendedly described while a complete mathematical model is proposed for the asset rating phase. The developed energy audit procedure was applied to poultry chambers located in lowland and mountainous areas of Epirus Greece for chambers of various sizes and technology levels. The energy intensity indices varied from 46 to 89 kWh/m2 of chamber area 0.25-0.48 kWh/kg of produced meat or 0.36-1.3 kWh/bird depending on the chamber technology level (insulation, automation, etc.) and the location where the unit was installed. The biggest energy consumer was heating followed by energy consumption for ventilation and cooling. An advanced technology level can improve energy performance by ~ 27%-31%. Proper insulation (4-7 cm) can offer a reduction of thermal energy consumption between 10 and 35%. In adequately insulated chambers, the basic heat losses are due to ventilation. Further energy savings can be achieved with more precise ventilation control. Automation can offer additional electrical energy saving for cooling and ventilation (15-20%). Energy-efficient lights can offer energy saving up to 5%. The use of photovoltaic (PV) technology is suggested mainly in areas where net-metering holds. The use of wind turbines is feasible only when adequate wind potential is available. Solar thermal energy is recommended in combination with a heat pump if the unit's heating and cooling systems use hot/cold water or air. Finally, the local production of biogas with anaerobic fermentation for producing thermal or electrical energy, or cogenerating both, is a choice that should be studied individually for each farm.

Untargeted UHPLC-MS metabolic profiling as a valuable tool for the evaluation of eggs quality parameters after dietary supplementation with oregano, thyme, sideritis tea and chamomile on brown laying hens.

INTRODUCTION: Bioactive constituents of medicinal-aromatic plants used as feed additives may affect the metabolic profile and oxidative stability of hen eggs. OBJECTIVES: To determine the effects of dietary supplementation with a mixture of dried oregano, thyme, sideritis tea and chamomile on laying hen performance, egg quality parameters, and oxidative stability in the egg yolk were monitored. METHODS: In this trial 432 hens were allocated in two treatments (unsupplemented vs. supplemented with the mixture) and fed for 42 days. Eggs were collected at the end of the trial period, egg yolk was separated, extracted, and the total phenolic content (TPC) and oxidative stability was measured. Furthermore, LC-MS metabolic profile of eggs was studied and pathway analysis was elaborated in MetaboAnalyst to facilitate annotation of features. RESULTS: Overall, egg production and feed conversion ratio were not affected by the supplementation. However, eggs from the supplemented treatment showed improved shell thickness and strength, and yolk resistance to oxidation. Moreover, LC-MS metabolomic analysis of egg yolk of supplemented and unsupplemented layers showed significant variations and tight clustering in unsupervised principal component analysis due to different chemical profiling of egg yolk. LC-MS study showed that secondary metabolites of aromatic plants did not transfer into yolk, nevertheless the feed supplementation impacted the pathway metabolism of tyrosine, phenylalanine, propanate, and the biosynthesis of aminoacyl-tRNA, phenylalanine, tyrosine and tryptophan. CONCLUSIONS: The dietary supplementation of layers with a mixture of dried medicinal aromatic plants affected shell thickness and strength, the lipid and protein oxidative stability and increased tyrosine and phenylalanine content in eggs.

Efficacy of a Dietary Polyherbal Formula on the Performance and Gut Health in Broiler Chicks after Experimental Infection with Eimeria spp.

One-hundred and fifty, one-day-old Ross-308 female chicks were randomly allocated to five equal treatments: NCONTR negative control-not challenged; PCONTR positive control-challenged; PHERB1 and PHERB2 diets were supplemented with phytogenic formula (1 and 2 g/kg feed, respectively)-challenged; PSALIN diet was supplemented with salinomycin (60 mg/kg feed)-challenged. Challenge was made by oral inoculation with 3.5 × 104 E. acervulina, 7.0 × 103 E. maxima and 5.0 × 103 E. tenella oocysts, at 14 days of age. One week post inoculation, bloody diarrhea, oocysts numbers, and intestinal lesions were evaluated, along with intestinal microbiota, viscosity, and pH of digesta, and histopathology. PHERB2 had a comparable (p ≤ 0.001) growth performance and feed conversion ratio to PSALIN. PHERB1 and PHERB2 had similar (p ≤ 0.001) oocyst counts to PSALIN and lower than PCONTROL. PHERB2 and PSALIN had lower (p ≤ 0.001) jejunal, ileal, and cecal lesion scores compared to PCONTR. PHERB1 and PHERB2 had higher (p ≤ 0.001) jejunal and cecal lactobacilli and lower (p ≤ 0.001) coliform counts compared to other treatments. PCONTR had lower (p ≤ 0.001) jejunum villus height, height to crypt ratio, and villus goblet cells. Breast and thigh meat resistance to oxidation was improved (p ≤ 0.001) in PHERB1 and PHERB2 compared to the PCONTR. The polyherbal formula exerted a substantial improvement on growth performance and intestinal health of the Eimeria-challenged birds.

Identification by MALDI-TOF MS and Antibiotic Resistance of Riemerella anatipestifer, 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.