Gene expression of pregnancy-associated glycoproteins-1 (PAG-1), interferon-tau (IFNt) and interferon stimulated genes (ISGs) as diagnostic and prognostic markers of maternal-fetal cellular interaction in buffalo cows.

The aim of this study was to determine the presence of Pregnancy-associated glycoprotein -1 (PAG-1) mRNA expression in the maternal circulation of pregnant buffaloes during the early stage of pregnancy. Contemporaneously, the mRNA expression levels of Interferon-tau (IFNt) and some Interferon stimulated genes (ISGs) (interferon stimulated gene 15 ubiquitin-like modifier interferon, ISG15; Mixoviruses resistance 1 and 2, MX1 and MX2; 2',5'-oligoadenylate synthase 1,OAS1) were evaluated in order to expand our knowledge of the molecular processes involved in the early stages of pregnancy and to identify potential biomarkers of maternal-fetal cellular interaction in buffalo. The study was conducted on 38 synchronized and artificially inseminated buffalo cows (d 0), divided ex post into 3 groups: Pregnant (n = 17), Non-pregnant (n = 15) and Embryo mortality (n = 6). Blood samples were collected on d 14, 19, 28 and 40 after artificial insemination (AI) for peripheral blood mononuclear cells (PBMCs) isolation. Expression levels of mRNA of PAG-1, IFNt, ISG15. MX1, MX2 and OAS1 were measured using RT-qPCR. No significant changes were observed in IFNt and PAG gene expressions between groups, while significant differences (p < 0.001) were found for ISG15, MX1, MX2, and OAS1. Pairwise comparisons revealed that the differences between groups occurred on days 19 and 28 post-AI. ISG15 proved to have the best diagnostic performance for distinguishing between pregnant animals and animals that experienced embryo mortality with the ROC analysis. According to the results of the univariate analyses, day 19 was identified as the most indicative to discriminate between groups while the most reliable genes for this differentiation were ISG15, MX1 and MX2. MX2 proved to be the best gene for discriminating pregnant buffaloes using the discriminant analysis, while MX1 was the gene that best predicted embryo mortality. Our results showed that among PAG-1, IFNt and ISGs expression as diagnostic and prognostic markers of maternal-fetal cellular interaction in buffalo cows, ISGs proved to be the best peripheral biomarkers for predicting pregnancy and embryonic mortality during the peri-implantation period. These insights into the mechanisms behind maternal-fetal interaction and the development of a method for the early detection of embryo distress may enable us to implement effective strategies to support embryo survival.

Dietary oregano (Origanum vulgare L.) aqueous extract improves oxidative stability and consumer acceptance of meat enriched with CLA and n-3 PUFA in broilers.

The effect of a dietary oregano aqueous extract on meat fatty acid profile, quality, and consumer acceptance in chickens fed a diet rich in polyunsaturated fatty acids (PUFA) was evaluated in 3 consecutive trials.For each trial, 171 day-old Ross 308 chicks were randomly divided in replicates of 19 birds each and assigned to one of 3 experimental diets: 1) basal control diet, 2) basal diet supplemented with 0.2 g/kg of oregano aqueous extract, and 3) basal diet supplemented with 150 ppm of vitamin E (as positive control). To better analyze the antioxidant activity of both oregano and vitamin E, all the experimental diets were enriched with a fatty acid supplement consisting in a mixture of PUFA at the same dose (1.16 %) in both starter and finisher feeds. Oregano supplementation positively influenced (P < 0.05) broiler live performance. No differences were observed in physicochemical or proximal composition or in total fatty acid composition of breast meat. Dietary oregano influenced meat composition in terms of total phenolic content, antioxidant capacity, and thiobarbituric acid-reactive substances, improving meat resistance to oxidation, compared to both other groups. During consumer tests, meat from the 3 dietary groups obtained the same liking score in a blind session. Under informed condition, consumer perception was positively influenced by labeling for all the considered attributes. Furthermore, consumers showed a higher expectation for meat derived from chickens fed the oregano extract when compared to that deriving from the other 2 groups.Results obtained in the present study allow stating that using oregano aqueous extract in diets enriched with PUFA can represent a valid solution to increase live weight of chickens, improve resistance to oxidation of meat, and positively influence consumer perception of poultry meat.

Dietary Lactobacillus acidophilus positively influences growth performance, gut morphology, and gut microbiology in rurally reared chickens.

In a market undergoing constant evolution, the production of chicken meat that consumers would perceive as "natural" and "animal friendly" is crucial. The use of probiotics in rurally reared chickens could represent a major opportunity to achieve mutual benefit for both the industry and consumers. A total of 264 male Kabir chicks were randomly distributed to one of 2 dietary treatments: the L group received a commercial feed supplemented with 2.0 g/100 kg of Lactobacillus acidophilus D2/CSL, while the C group received the same basal diet without the additive. To assess the effects of probiotic supplementation in the chickens' diet, productive performance was evaluated at d 21 and 42, whereas microbiological analyses of the intestinal content and intestinal histology and morphometry were performed at the end of the trial (d 42). At d 21 and 42, L birds showed better (P < 0.001) performance in terms of body weight, average daily gain, and feed conversion ratio. Enterococci, staphylococci, and Escherichia coli populations were not influenced by dietary treatment. On the contrary, Lactobacillus population increased (P = 0.032) in the L group. Furthermore, a tendency (P = 0.069) was observed for the coliforms to be influenced by diet, with lower values in the L group in comparison to the C group. Histological techniques revealed that the number of goblet cell containing neutral mucins was lower in the C group. Morphometric evaluations demonstrated that the probiotic supplementation increased the height of the mucosal layer by improving (P = 0.040) villus height, while crypt depth was unaffected. In conclusion, the results obtained in this study demonstrate that it is possible to use Lactobacillus acidophilus D2/CSL (CECT 4529) in rurally reared chicken breeds with positive effects on performance and gut health.

Dietary integration with oregano (Origanum vulgare L.) essential oil improves growth rate and oxidative status in outdoor-reared, but not indoor-reared, pigs.

The effects of a diet supplemented with oregano essential oil on performance, oxidative status, pork quality traits and sensorial properties were evaluated. In two studies, 72 pigs in indoor or outdoor conditions were assigned to either a control diet or an identical diet supplemented with 0.2% oregano essential oil. Pigs reared outdoor showed lower live weight, average daily gain and average gain:feed ratio compared to indoor pigs. The oregano supplementation improved the growth performance of the outdoor- but not the indoor-reared animals. The serum oxidative status was influenced by the diet. A higher oxidative stability was observed in the oregano-supplemented groups. As for the rearing conditions, the data suggest that after an initial adapting period, the free-range farming systems could be better tolerated by pigs. Meat derived from pigs reared outdoor showed higher pH and a* values. Lightness was influenced by both the diet and the rearing conditions. The control group reared indoor showed shear force values higher than both supplemented groups, while no differences were detected with the control group reared outdoor. In the consumer test performed under blind conditions, the oregano groups achieved higher consistency scores compared with the control. Under informed conditions, the meat derived from the oregano-supplemented pigs reared outdoor received the highest scores for consistency and overall liking regardless of the rearing system. The same result for the overall liking score was obtained in the expectation test. The data obtained showed that dietary oregano essential oil can be effective in reducing performance losses due to the outdoor-rearing system, increasing the oxidative status of the animal and oxidative stability of the meat, without modifying the meat quality traits and improving consumer perceptions of the meat quality.

Gut complex carbohydrates and intestinal microflora in broiler chickens fed with oregano (Origanum vulgare L.) aqueous extract and vitamin E.

One hundred and seventy one-day-old female broiler chicks were randomly divided into three groups fed with different dietary treatments: basal control diet (C); C supplemented (2 g/kg) with an oregano aqueous extract (O); C supplemented (150 mg/kg) with vitamin E (E). Growth performance was evaluated at 21 (T1) and 42 days (T2). On the same days, morphological, histochemical and microbiological analyses were performed. The O group showed the highest (p < 0.01) body weight at T1, while no differences were observed at T2. Light microscopic observation and conventional histochemistry showed no differences with regard to the two sampling times, whereas significant differences emerged among the treatments. The O treatment generally enhanced goblet cell reactivity more than both the C and E treatments. Coliform count was lower in the ileum tract of the O group at both T1 and T2 (p < 0.05) and increased with age in all groups. Escherichia coli showed the lowest values in the caecum of the O group (p < 0.001) at both sampling times. Enterococci, lactobacilli and staphylococci populations showed no differences among the different experimental groups in the caecum. In the ileum, the O group did not exhibit the sharp decline (p < 0.001) in the lactic acid bacteria population observed in the other two experimental groups. In conclusion, oregano aqueous extract supplementation seemed to elicit the best response among treatments, enabling better growth performance, enhancing both the quantity and quality of glycoconjugates involved in indirect defence actions and significantly reducing both the coliform and E. coli counts.

Effects of two different probiotics on microflora, morphology, and morphometry of gut in organic laying hens.

The current study investigated the effects of Lactobacillus acidophilus and Bacillus subtilis, used as probiotics, on the microflora, morphology, and morphometry of the gut in organic laying hens. The birds (180 Hy-Line laying hens) were divided into 3 homogenous groups and received a pre-deposition diet from 16 to 20 wk of age and a deposition diet for the remaining 7 months of the experiment. The control group ( CTR: ) was fed a corn-soybean cake-based diet, the second group ( L: ) received the same diet supplemented with 0.1% of L. acidophilus while in the third group ( B: ) the basal diet was supplemented with 0.05% of B. subtilis At 18 wk of age ( T1: ) and at 5 ( T2: ) and 7 months ( T3: ) from the beginning of deposition, 9 subjects per group were humanely killed for microbiological, morphological and morphometric analyses of the intestinal tract. The 2 probiotic-supplemented diets increased Lactobacillus spp. and Bifidobacterium spp. counts compared with the CTR diet. The lowest viable counts of E. coli, coliforms and staphylococci were observed in the L group (P < 0.001). Clostridium spp. decreased (P < 0.001) in both L and B subjects. The probiotic supplementation appeared to affect the intestinal microbial population, promoting the presence of beneficial bacteria such as Lactobacillus spp. and Bifidobacterium spp. and reducing potential harmful bacteria such as E. coli, clostridia and staphylococci. Morphological and morphometric analyses did not reveal substantial differences among groups. At T3, the plasma cell infiltrate in the villi of the CTR hens was more severe than that observed in the L and B groups (P = 0.009).

Effects of dietary Lactobacillus acidophilus and Bacillus subtilis on laying performance, egg quality, blood biochemistry and immune response of organic laying hens.

The objective of this study was to evaluate the effects of two different probiotic micro-organisms on the performance, egg quality and blood parameters of organically reared hens. A total of 900 16-week-old Hy-Line layer hybrids were randomly assigned to three groups of 300 birds each. The control (CTR) group was fed a corn-soya bean cake-based diet; the L group was fed the same diet supplemented with 0.1% Lactobacillus acidophilus, while the B group was fed the same diet supplemented with 0.05% Bacillus subtilis. Data were recorded at the beginning (weeks 5 and 6: T1) and at the end (weeks 19 and 20: T2) of the experiment, and no differences in hen performance were recorded between dietary groups or sampling times. All of the investigated clinical chemistry parameters, except GGT, were affected by diet (p < 0.05), with the best results recorded for the probiotic-treated groups. The immune-response values showed higher blood bactericidal activity in the B and L groups at T2 (p < 0.05) and a lower lysozime concentration in the B group at T1. Higher antibody production against Newcastle disease virus was observed in the L group compared to the CTR (p = 0.013). No differences in oxidative status were recorded, and no effects of diet on egg quality were observed. Among the physical egg characteristics, only the Roche scale colour was affected by diet (p < 0.05): the egg yolk was paler in the L group. The age of the hen was the most relevant factor affecting physical egg characteristics. The chemical parameters of the egg were almost unaffected by supplementation with probiotics except for the lipid content, which decreased with the L diet (p < 0.05). Both probiotic inclusions had beneficial effects on hen metabolism and welfare, and L. acidophilus induced the best immune response.

Dietary effects of a mix derived from oregano (Origanum vulgare L.) essential oil and sweet chestnut (Castanea sativa Mill.) wood extract on pig performance, oxidative status and pork quality traits.

The effects of a pre-formulated commercial plant extract mix, composed of equal parts of oregano essential oil and sweet chestnut wood extract, on performance, oxidative status and pork quality traits were evaluated. In two 155-d studies, 60 pigs (mean liveweight: 42.9 kg) were assigned to either a control diet (CTR) or an identical diet supplemented (0.2%) with the plant extract mix (OC). No differences in the growth rate were observed. Glutathione peroxidase and glutathione reductase activities in the OC muscles (Longissimus lumborum) were higher than in CTR muscles. The lipid oxidation of meat was lower in the OC group. In the cooked meat samples, OC animals had the lowest L* and H° values and the highest a* values. The OC meat received higher scores for colour, taste and overall liking in both the blind and the labelled consumer tests.

Effects of field bean (Vicia faba L. var. minor) dietary supplementation on plasma thyroid hormones, insulin, insulin-like growth factor-1 concentrations and mohair characteristics in growing Angora goat kids.

Twenty-two Angora kids were used to study the effect of a dietary supplementation with field bean (Vicia faba L. var. minor) on hair follicle activity, mohair characteristics and plasma total thyroxine (T4), triiodothyronine (T3), insulin (INS) and insulin-like growth factor-1 (IGF-1) concentrations. At birth, their mothers were divided in two groups (S: supplemented, C: control), the S group being supplemented with 300 g/head/day of whole field bean. At weaning, diet of kids from S group was supplemented with 80 g/head/day of whole field bean. At weaning, secondary hair follicle activity (S: 0.91 ± 0.01, C: 0.84 ± 0.02, p < 0.01) and staple length (S: 5.82 ± 0.13 cm, C: 5.16 ± 0.14 cm, p < 0.001) were greater in the S group. At 155 days of age, secondary fibre diameter (S: 16.54 ± 0.35 µm, C: 18.09 ± 0.31 µm, p < 0.01) was higher in the C kids. Concentrations of total plasma T4 and T3 were higher (p < 0.001) in S (120 ± 12 and 4.87 ± 0.92 ng/ml respectively) than in C kids (92 ± 8 and 2.97 ± 0.77 ng/ml respectively). At weaning, T4 values were negatively correlated (p < 0.05) with the secondary fibre diameter and the T3/T4 ratio was positively correlated (p < 0.05) with the secondary to primary hair follicle ratio. At 155 days of age, both T3 and T3/T4 values were positively correlated (p < 0.01) with staple length. Plasma INS (0.11 ± 0.03 ng/ml) and total IGF-1 concentrations (232 ± 13 ng/ml) were not affected by diet. Improvements of diet quality in kids positively affected fibre diameter and length and follicle activity rate. We suggest that increased circulating thyroid hormones concentrations, but not those of INS and IGF-1, may contribute to the beneficial actions of nutritional supplementation on anatomical growth and mohair production of kids.

In vivo mechanical and in vitro electromagnetic side-effects of a ruminal transponder in cattle.

This work was undertaken to assess the long-term impacts of a ruminal transponder, used for electronic identification, on ruminal motility and on health and performance of cattle, as well as to study the electromagnetic effects on ruminal bacteria in vitro. A passive transponder (51.4 g, 67 x 17 mm) was delivered into the forestomachs of 8 calves, 32 bulls, 10 heifers, and 40 dairy cows. Final readability was 87.5% in calves, 96.9% in bulls, 90% in heifers, and 100% in cows at 481, 360, 650, and 601 d, respectively, after transponder administration. The transponder did not affect production or reproduction of cows over a 2-yr period, or performance of bulls, or mortality compared with control animals. Chewing movements per bolus were lower (P <0.01) in treated animals than in controls (49.6 vs. 52.2, 51.2 vs. 63.6, and 57.0 vs. 59.7 for bulls, heifers, and cows, respectively). Regurgitation frequency (number of boluses/10 min) tended to be greater in treated cattle: 12.4 vs. 11.3 (P = 0.07), 11.3 vs. 10.6, and 11.3 vs. 10.7 (P = 0.08) for bulls, heifers, and cows, respectively. Rumination patterns of calves fitted with transponders within the first weeks of life were similar to controls. During the experiment, 43 treated animals (8 calves, 29 bulls, and 6 cows) were slaughtered. Thirty transponders were localized in the reticulum (3 calves, 24 bulls, and 3 cows), 11 in the rumen (4 calves, 4 bulls, and 3 cows), and 2 were not recovered (1 calf and 1 bull). Within the calves, 57% of the boluses were found in the rumen. In 8 reticula (2 calves and 6 bulls) and 1 rumen (1 cow), an impression left by physical contact of the transponder was observed, although histological examination did not reveal specific lesions in the mucosa of the dystrophic areas. In strained, whole ruminal contents incubated in vitro, pH values were lower after 24 and 48 h (P <0.001) of continuous exposure to an electromagnetic field induced by the transponder-reading system. After 48 h of incubation, total bacterial numbers and NH3-N concentration were greater (P <0.001) in exposed flasks than in controls. These data indicate that the transponder may alter, via mechanical action, the reticuloruminal mucosa and rumination patterns. Furthermore, the transponder may increase, via its electromagnetic action, the growth rate and metabolic activity of ruminal bacteria.

Tenderization of wether lambs meat through pre-rigor infusion of calcium ions.

A study involving 36 wether lamb carcasses was conducted in order to evaluate the effects of 0.3 M CaCl(2) injection on final tenderness in muscle Longissimus thoracis et lumborum. Injection of wether lamb carcasses with CaCl(2) accelerated the post mortem tenderization process. Both control and distilled water injected animals had similar Ca(2+)-dependent proteases (µ-calpain and m-calpain) and their inhibitor (calpastatin) activities, whereas these were all significantly decreased in CaCl(2) injected animals. The effect of CaCl(2) injection on tenderness was recorded by measuring shear force values 2 and 6 days post mortem. Tenderness was significantly improved by CaCl(2) injection at both at 2 and 6 days post mortem. It was concluded that activation of µ-calpain and m-calpain was responsible for the observed post mortem proteolysis and tenderization.