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Computational fluid dynamics (CFD) may aid the design of barn ventilation systems by simulating indoor cattle thermal welfare. In the literature, CFD models of mechanically and naturally ventilated barns are proposed separately. Hybrid ventilation relies on cross effects between air change mechanisms that cannot be studied using existing models. The objective of this study was to develop a CFD methodology for modelling animal thermal comfort in hybrid ventilated barns. To check the capability of CFD as a design evaluation tool, a real case study (with exhaust blowers) and an alternative roof layout (with ridge gaps) were simulated in summer and winter weather. Typical phenomena of natural and mechanical ventilation were considered: buoyancy, solar radiation, and wind together with high-speed fans and exhaust blowers. Cattle thermal load was determined from a daily animal energy balance, and the assessment of thermal welfare was performed using thermohygrometric indexes. Results highlight that the current ventilation layout ensures adequate thermal welfare on average, despite large nonuniformity between stalls. The predicted intensity of heat stress was successfully compared with experimental measurements of heavy breathing duration. Results show strong interactions between natural and mechanical ventilation, underlining the need for an integrated simulation methodology.

In this work, milk samples collected in a cohort of intensive dairy farms of the Po Valley (Italy) were screened for their (poly)-phenolic profile to check the occurrence of phenolic metabolites of biological interest. The selected dairy farms were previously classified on the basis of their cow feeding system, considering the utilization of corn silage as the main ingredient of the rations. Overall, ultra-high-pressure liquid chromatography coupled with mass spectrometry using an Orbitrap analyzer, followed by unsupervised and supervised statistics, allowed identifying clear different phenolic distributions in the milk samples. Accordingly, a great variability in the phenolic profiles of the different milk samples was observed, with two main phenolic clusters outlined by the unsupervised hierarchical clustering approach and not fully correlated to the nutritional strategy considered. The variables’ importance in the projection approach allowed selecting the most important metabolites, resulting in samples’ discrimination. Among the most discriminative compounds, we found phenolic metabolites (such as hippuric acid and 4-hydroxyhippuric acid), followed by lignans (such as enterolactone) and isoflavonoids (such as equol and O-desmethylangolensin). Taken together, our findings suggested that both the feeding systems and the ability of dairy cows to process parent phenolic compounds were the main factors providing the final (poly)-phenolic profile of the milk samples. Future targeted and ad hoc studies appear of great interest to evaluate the potential biological effects of these compounds on cow health.

A survey regarding crop enterprise management, forages cost of production, dairy cattle management including reproductive management, housing, heat abatement, body condition scoring, nutrition, grouping strategies, and income over feed cost performance, was carried out from December 2016 to January 2018 on 50 dairy farms by the Department of Animal Science, Food and Nutrition of Università Cattolica del Sacro Cuore (Piacenza, Italy). A total of 41 herds (82%) completed the survey. Average herd size was 327 ± 162 lactating cows with the average land size of 160 ± 94 ha per farm. Herds were located in the provinces of Cremona (17), Brescia (8), Mantova (7), Piacenza (5), Cuneo (4), Bergamo (3), Lodi (3), Torino (2), and Venezia (1). These farms sold 32.8 ± 2.01 kg of milk/day per cow, had an annual culling rate of 34.0 ± 4.00%, a calving interval of 14.16 ± 0.58 months., and a 21-days pregnancy rate of 17.05 ± 2.58%. Implementing effective management strategies to contrast the damage caused by Ostrinia nubilalis, Diabrotica spp. and Myocastor coypus were identified as the main crop enterprise challenges. Main forages cultivated were alfalfa and corn silage second seeding with a total cost of production of (€/ha) 1968 ± 362 and 2,581 ± 221, with an average yield of 9.61 ± 1.24 and 17.22 ± 2.46 ton of DM per hectare, respectively. Results of this study can provide useful benchmark or reference for dairy management practices, crops and dairy performances, forages production costs on very well-managed North Italian dairy farms at the present time.Highlights benchmarks for dairy farms management practices, economic and reproductive performance cost of production of forages in northern Italy

In this work, a retrospective screening based on ultra-high-performance liquid chromatography (UHPLC) coupled with high-resolution mass spectrometry (HRMS) based on Orbitrap-Q-Exactive Focus™ was used to check the occurrence of regulated and emerging mycotoxins in bulk milk samples. Milk samples were collected from dairy farms in which corn silage was the main ingredient of the feeding system. The 45 bulk milk samples were previously analyzed for a detailed untargeted metabolomic profiling and classified into five clusters according to the corn silage contamination profile, namely: (1) low levels of Aspergillus- and Penicillium-mycotoxins; (2) low levels of fumonisins and other Fusarium-mycotoxins; (3) high levels of Aspergillus-mycotoxins; (4) high levels of non-regulated Fusarium-mycotoxins; (5) high levels of fumonisins and their metabolites. Multivariate statistics based on both unsupervised and supervised analyses were used to evaluate the significant fold-change variations of the main groups of mycotoxins detected when comparing milk samples from clusters 3, 4, and 5 (high contamination levels of the corn silages) with cluster 1 and 2 (low contamination levels of the corn silages). Overall, 14 compounds showed a significant prediction ability, with antibiotic Y (VIP score = 2.579), bikaverin (VIP score = 1.975) and fumonisin B2 (VIP score = 1.846) being the best markers. The k-means clustering combined with supervised statistics showed two discriminant groups of milk samples, thus revealing a hierarchically higher impact of the whole feeding system (rather than the only corn silages) together with other factors of variability on the final mycotoxin contamination profile. Among the discriminant metabolites we found some Fusarium mycotoxins, together with the tetrapeptide tentoxin (an Alternaria toxin), the α-zearalenol (a catabolite of zearalenone), mycophenolic acid and apicidin. These preliminary findings provide new insights into the potential role of UHPLC-HRMS to evaluate the contamination profile and the safety of raw milk to produce hard cheese.

In this study, an untargeted metabolomics approach based on ultra-high-performance liquid chromatography coupled with high-resolution mass spectrometry (UHPLC-HRMS) was used for investigating changes in chemical profiles of cow milk considering diets based on mycotoxins-contaminated corn silages. For this purpose, 45 milk samples were classified into five clusters according to the corn silage contamination profile, namely (1) low levels of Aspergillus- and Penicillium-mycotoxins; (2) low levels of fumonisins and other Fusarium-mycotoxins; (3) high levels of Aspergillus-mycotoxins; (4) high levels of non-regulated Fusarium-mycotoxins; (5) high levels of fumonisins and their metabolites, and subsequently analyzed by UHPLC-HRMS followed by a multivariate statistical analysis (both unsupervised and supervised statistical approaches). Overall, the milk metabolomic profile highlighted potential correlations between the quality of contaminated corn silages (as part of the total mixed ration) and milk composition. Metabolomics allowed to identify 628 significant milk metabolites as affected by the five levels of corn silage contamination considered, with amino acids and peptides showing the highest metabolite set enrichment (134 compounds). Additionally, 78 metabolites were selected as the best discriminant of the prediction model built, possessing a variable importance in projection score >1.2. The average Log Fold-Change variations of the discriminant metabolites provided evidence that sphingolipids, together with purine and pyrimidine-derived metabolites were the most affected chemical classes. Also, metabolomics revealed a significant accumulation of oxidized glutathione in milk samples belonging to the silage cluster contaminated by emerging Aspergillus toxins, likely involved in the oxidative imbalance. These preliminary findings provide new insights into the potential role of milk metabolomics to provide chemical indicators of mycotoxins-contaminated corn silage feeding systems.

The chemical composition of milk can be significantly affected by different factors across the dairy supply chain, including primary production practices. Among the latter, the feeding system could drive the nutritional value and technological properties of milk and dairy products. Therefore, in this work, a combined foodomics approach based on both untargeted metabolomics and metagenomics was used to shed light onto the impact of feeding systems (i.e., hay vs. a mixed ration based on hay and fresh forage) on the chemical profile of raw milk for the production of hard cheese. In particular, ultra-high-performance liquid chromatography quadrupole time-of-flight mass spectrometry (UHPLC-QTOF) was used to investigate the chemical profile of raw milk (n = 46) collected from dairy herds located in the Po River Valley (Italy) and considering different feeding systems. Overall, a total of 3320 molecular features were putatively annotated across samples, corresponding to 734 unique compound structures, with significant differences (p < 0.05) between the two feeding regimens under investigation. Additionally, supervised multivariate statistics following metabolomics-based analysis allowed us to clearly discriminate raw milk samples according to the feeding systems, also extrapolating the most discriminant metabolites. Interestingly, 10 compounds were able to strongly explain the differences as imposed by the addition of forage in the cows’ diet, being mainly glycerophospholipids (i.e., lysophosphatidylethanolamines, lysophosphatidylcholines, and phosphatidylcholines), followed by 5-(3′,4′-Dihydroxyphenyl)-gamma-valerolactone-4′-O-glucuronide, 5a-androstan-3a,17b-diol disulfuric acid, and N-stearoyl glycine. The markers identified included both feed-derived (such as phenolic metabolites) and animal-derived compounds (such as lipids and derivatives). Finally, although characterized by a lower prediction ability, the metagenomic profile was found to be significantly correlated to some milk metabolites, with Staphylococcaceae, Pseudomonadaceae, and Dermabacteraceae establishing a higher number of significant correlations with the discriminant metabolites. Therefore, taken together, our preliminary results provide a comprehensive foodomic picture of raw milk samples from different feeding regimens, thus supporting further ad hoc studies investigating the metabolomic and metagenomic changes of milk in all processing conditions.

Lectins, phytic acid and condensed tannins exert major antinutritional effects in common bean when grains are consumed as a staple food. In addition, phaseolin, i.e., the major storage protein of the bean seed, is marginally digested when introduced in the raw form. Our breeding target was to adjust the nutrient/antinutrient balance of the bean seed for obtaining a plant food with improved nutritional value for human consumption. In this study, the seeds of twelve phytohaemagglutinin-E-free bean lines carrying the mutations low phytic acid, phytohaemagglutinin-L-free, α-Amylase inhibitors-free, phaseolin-free, and reduced amount of condensed tannins, introgressed and differently combined in seven genetic groups, were analyzed for their nutrient composition. Inedited characteristics, such as a strong positive correlation (+0.839 **) between the genetic combination “Absence of phaseolin + Presence of the α-Amylase Inhibitors” and the amount of “accumulated iron and zinc”, were detected. Three lines carrying this genetic combination showed a much higher iron content than the baseline (+22.4%) and one of them in particular, achieved high level (+29.1%; 91.37 µg g−1) without any specific breeding intervention. If confirmed by scientific verification, the association of these genetic traits might be usefully exploited for raising iron and zinc seed content in a bean biofortification breeding program.

The increasing use in Italy of the Cornell Net Carbohydrate and Protein System (CNCPS) led researchers of five Italianuniversities to carry out a collaborative study to evaluate the precision of the CNCPS chemical analysis and derivate fractions.Each laboratory conducted in duplicate the chemical analyses according to the Weende (dry matter; crude protein;ether extract; crude fibre; ash), Van Soest (neutral and acid detergent fibre, NDF and ADF; acid detergent lignin; ADL)and CNCPS (soluble proteins, SP; non-protein nitrogen, NPN; neutral and acid detergent insoluble protein, NDIP andADIP; starch, ST) schemes on the same five feeds (barley meal, wheat straw, maize silage, dried lucerne and fieldbeans). Anomalous analytical data were identified and corrected by the “box-plot” graphic tool before the calculation ofthe CNCPS protein (B1, B2 and B3) and carbohydrate (A, B2 and C) fractions. Finally, repeatability (chemical analysis) andreproducibility (chemical analysis and fractions) were calculated and expressed as relative values (repeatability andreproducibility standard deviation as percentage of the corresponding mean, RSDr and RSDR, respectively). Chemicalanalyses of the Weende scheme, together with NDF, ADF and ST analyses, have satisfactory repeatability (0.3-6.2%) andreproducibility (0.3-11.2%) values. On the contrary the ADL, NPN, NDIP and ADIP analyses showed high variability, bothwithin and between laboratories (RSDr and RSDR between 20 and 45%). The SP analysis had an intermediate value ofprecision (RSDr=10.6%; RSDR=16.4%).Finally, since different combinations of several chemical analyses with scarce (ADL, NPN, NDIP, ADIP, SP) or average precision(e.g. NDF and starch) are used to calculate CNCPS fractions (excluding B2 protein fraction), also the reproducibilitiesof these fractions are poor and range from 10 to 20%.

The objective of the experiment was to monitor plasma levels of aflatoxin B1 (AFB1), B2 (AFB2), G1 (AFG1), G2 (AFG2) and M1 (AFM1) in lactating dairy cows fed a single oral bolus with aflatoxin naturally contaminated corn meal (Trial 1). The possible aflatoxins (AFs) absorption through mucous membranes was also investigated using the vaginal mucosa (Trial 2). In trial 1, seven lactating Holstein dairy cows were given a single oral bolus of a naturally contaminated corn meal assuring an intake of 4.89 mg AFB1, 1.01 mg AFB2, 10.63 mg AFG1 and 0.89 mg AFG2. Blood samples were collected at 0 and 5, 10, 15, 20, 25, 30 minutes after treatment. In trial 2 an aflatoxin dosage similar to that of trial 1 was provided through vaginal implant to eight lactating Holstein dairy cows. Blood samples were collected at 0 and 15, 30, 60, 180, 360 minutes after treatment. Individual milk samples of six milkings, one before and five after treatment, were also collected. Plasma and milk samples were analysed by HPLC for AFB1, AFB2, AFG1, AFG2 and AFM1 contents. In trial 1 AFB1 in plasma peaked (33.6 ng/L) as soon as 20 minutes after treatment. The plasma AFM1 was already detectable at 5 minutes (10.4 ng/L) and peaked at 25 minutes (136.3 ng/L). In trial 2 only AFB1 and AFM1 were detectable in plasma, starting from the first sampling time (15 minutes), with values of 10.7 and 0.5 ng/L, respectively. The AFB1 peaked at 30 minutes (23.9 ng/L). The AFB1 excreted in milk as AFM1 had the highest concentration (203.0 ng/L) in the first milking after treatment and decreased close to the starting values after 36 hours from treatment. The prompt appearance of studied aflatoxins, and their metabolites, in plasma suggests absorption might also take place in mouth or oesophageal mucous membranes, before the rumen compartment. Results support the hypothesis that the cytochrome P450 oxidative system, which is present in these tissues and in leukocytes, could be involved in the conversion of the AFB1 in AFM1. The absorption of AFB1 through the vaginal mucosa confirms the passive diffusion as a probable mechanism for AFB1 absorption.

An assessment was made of the effect that different treatments (toasting, expansion, extrusion) have on the nutritionalvalue of protein plants (pea, faba bean, lupin). In a randomized block design, feeds were screened for enzymaticdigestibility of starch and protein, N solubility and in vitro protein degradability. Expansion and extrusion cause increasedstarch enzymatic degradability while toasting produced virtually no effects. In peas this value increased from 11.80% inmeal to 39.70% in the extruded product; 85.37% is the percentage for the expanded product, while 10.90% is the starchdigestibility value for toasted peas. In faba beans the extrusion process increased starch digestibility from 11.39% to85.05%, while in extruded lupins a complete starch hydrolysis was obtained, while in the meal the polysaccharide digestionwas 54.48%.The expansion and extrusion processes significantly decreased rumen degradability during the first 8 hours of incubation.Toasted peas had lower degradability if compared with controls but not with the other treatments. The onlypotentially alternative source to soybean is the extruded faba bean. In spite of its lower protein content, this feed ischaracterized by a considerably lower in vitro protein degradability than soybean. This implies that the digestible foodprotein content is comparable (124.90 g/kg DM) to that of soybean (109.78 g/kg DM) and definitely higher than thatof all other protein plants.

Four hybrids of class FAO 700: EA6601 and Simeto (Monsanto), Eleonora (Pioneer), Pampero (Syngenta), grown under identical conditions, underwent analysis to determine their analytical and botanical composition and DM (IVDMD) and NDF (IVNDFD) in vitrodigestibility. The content of fibrous fractions increased with maturity. For stalks there was an increase from an average value of 47.72% (V8) up to over 61% during the dough stage of the grain (R4) reaching over 73% during the phase of physiological maturation (R6). At the eighth-leaf vegetative stage in hybrids the stalk represent- ed 61.59% of the DM of the plant and reached 40.35% in the dough stage, while the leaf apparatus fall from 38.41% of the eighth-leaf stage DM to 18.19% during the dough stage of the grain and only 9.17% during physiological maturation. The ear, which represented only 12.93% of DM plant at the silking stage, increased to 41.47% and 53.01 respectively at the dough and physiological phases. At the dough stage (R4) the hybrids Simeto and Pampero, had a stalk percentage significantly higher (41.67% and 42.34% respectively; P< 0.01) compared with Eleonora (38.97%) and EA6601 (38.44%). Eleonora hybrid had a leaf proportion of 19.80% - higher than Pampero (17.93%), Simeto (17.60%), EA6601 (17.43%). This greater leafiness occurred with a lower incidence of the stalk, which has the highest loss of digestibility during maturing. The grain percentage of EA6601 was significantly higher than Pampero (26.46%), Simeto (27.33%) and Eleonora (27.88%). In the R4 phase the hybrid EA6601 had a higher stalk fibre content (ADF 41.38%; P< 0.05) compared to Pampero (37.61%) Simeto (38.14%) and Eleonora (39.16%). The effect of the phenological stage on the fiber content of the leaves was less evident, only the ADL increased from values of about 5% in the first period, to 8% and more in the final three stages. Even here, in dough stage, EA6601 differed (P< 0.01) from the others for its higher levels of ADL (9.35%) and ADF (35.05%), while the hybrids Eleonora and Pampero presented lower values of ADL (P 0.01) and ADF (7.90%). Simeto had the lowest ADF content (31.39%; P< 0.01). In the dough stage there was no significant differ- ences for in vitrodigestibility of DM and NDF between the hybrids - for all the botanical parts. In the whole plant, as maturity advanced, the differences of in vitrodigestibility of stalks, were counterbal- anced by the increasing proportion of grain.

The effect of extrusion of pea seeds (Pisum sativum) (PS), faba bean (Vicia faba, variety minor) (FB) and lupin seeds (Lupinus albus, variety multitalia) (LS) on broiler performance were evaluated. Four hundred sixty two 1d-old Ross male chicks, Marek vaccinated, were randomly assigned to seven dietary treatments (3 pens per treatment/22 birds per pen). Chicks were floor housed, ad libitum fed isocaloric and isonitrogenous diets and had free access to water. Artificial light was provided 10 h/d. The bulk of the base diet (control diet) was corn (48.8%, 53.7% and 57%), solvent-extracted soy- bean meal (42.8%, 37.3% and 33.4%), corn oil (4.4%, 5.2% and 6.3%), plus synthetic amino acids, minerals, trace minerals and vitamins, respectively for the 1-10d-old, 11-28d-old and 29 to 42d-old growing periods. The amounts of PS, FB and LS used on an as fed basis were: PS and extruded PS (EPS): 353 (1-10d-old), 356 (11-28d-old) and 350 (29- 42d-old) g/kg; FB and extruded FB (EFB): 479 (1-10d-old), 497 (11-28d-old) and 500 (29-42d old) g/kg; LS and extrud- ed LS (ELS): 360 (1-10d-old) and 300 (11-42d-old) g/kg. High levels of pea (350 g/kg) and faba bean (500 g/kg) did not show negative effects on body weight gain (BWG) and bird feed intake compared to control. Lupin at the 300 g/kg level reduced (P< 0.05) the BWG during the finishing period (22 to 42 d), however the effect disappeared over the whole experimental period (1-42 d) compared to the control group. The ELS group had a lower (P< 0.01) feed intake com- pared to the control group and to the LS group. The feed conversion rate (FCR) was similar among groups for the whole experimental period; however during the grower period the FCR was higher (P< 0.05) for the PS, FB and EFB groups com- pared to the control group. Birds consuming the PS diet had a reduced (P< 0.05) eviscerated carcass yield compared to the control group. The breast meat percent yield was higher (P< 0.01) for birds consuming the FB and EFB diets compared to the control group. There were no statistical differences in percent yield of the leg quarters and in blood parameters.

The ban of the meat and bone meal for entering animal diets and the concern of transgenic feeds poses a challenge toanimal nutritionists in Europe. The challenge is to find homegrown protein-rich feedstuffs, making sure no antinutritionalfactors are present which could interfere in the animals’ performance. The raw Pea (Pisum sativum) (RP), raw Fababean (Vicia faba, variety minor) (RFb) and raw Lupin (Lupinus albus, variety multitalia) (RL) were evaluated as alternativeprotein sources into broiler diets. Six hundred thirty 1d-old Ross male chicks, Marek vaccinated, were randomlyassigned to seven dietary treatments (5 pens per treatment/18 birds per pen). Chicks were floor housed, ad libitum fedisocaloric and isonitrogenous diets and had free access to water. Artificial light was provided 10 h/d. The bulk of the basediet (control diet) was corn (48.7%, 56.6% and 57%), solvent-extracted soybean meal (42.8%, 37.3% and 33.4%), cornoil (4.4%, 5.2% and 6.3%), plus synthetic amino acids, minerals, trace minerals and vitamins, respectively for the 1-10d-old, 11-28d-old and 29 to 42d-old growing periods. The RP, RFb and RL entered diets in substitution of the soybeanand corn according to the cost optimization (P100, Fb100 and L100, respectively for RP, RFb and RL) and at half of theoptimized quantity (RP50, RFb50 and RL50, respectively for RP, RFb and RL). The amount used as fed basis for the higherlevel of inclusion were: P100: 350 g/kg for all diets; Fb100: 480 g/kg (1-10d-old) and 500 g/kg (11-42d-old); L100:360 g/kg (1-10d-old) and 300 g/kg (11-42d-old). The average daily gain (ADG) were lower (P < 0.05) in the RP groupcompared to the control group. Over the whole period of growth, the RFb group had similar ADG compared to the controlgroup and for both levels of inclusion, whereas reduced (P < 0.05) ADG were observed in the RL100 group. Reduced(P < 0.05) ADG were also observed for the RFb100 and the RL100 groups when calculated over the first three weeks ofgrowth. Birds performance was improved (P < 0.05) in the RL50 group. No effects were observed on dressing percentageand breast and leg quarter cuts. The RFb and RL could represent valuable protein feeds in broilers diet formulation.

The 42 days trial was carried out using 140 piglets weaned at 28 days of age. The piglets were allocated according to weight and sex to the 5 dietary treatments with 7 replicates for each treatments (4 pens x 4 castrated males and 3 pens x 4 females). The piglets were fed according to the following experimental design: 1) control diet (CTR) with soybean meal (SBM) 44% c.p. as protein source; 2) CRT diets with 200 g/kg of raw pea (Pisum sativum) (RP); 3) CTR diet with 200 g/kg extruded pea (EP); 4) CRT diet with 170 g/kg raw lupin (Lupinus albusvar. Multitalia) (RL); 5) CTR diet with 170 g/kg of extruded lupin (EL). During the trial, animals were weighed at 0 - 21 and 42 days from the start of the trial. Feed intake was monitored and feed conversion ratio was calculated for the periods 0-21 d and 22-42 d. At the end of the trial, blood samples were taken for 14 animals for each dietary treatment (2 animals per replicate) and analysed for total protein, urea and liver activity (ALT, AST and ALP parameters). Average daily weight gain and feed intake did not differ according to dietary treatments whereas during the total experimental period (0-42 d), feed conversion ratio was higher for EP vsCTR diet (2.35 vs2.09, respectively; P compared with diets containing the raw ingredients did not differ. Feed conversion ratio for the RP was numerically high- er than for the EP (2.35 vs2.16 and 2.76 vs2.32, respectively during 22-42 d and 0-42 d periods). Blood parameters did not show significant difference among dietary treatments except for higher total protein for CTR diet vsRL diet, EL and RP (67.3 vs62.2, 62.8 and 63.6 g/l, respectively; PvsRL and RL (4.7 vs3.7 and 3.8 mmol/l respectively; P

Four wethers housed in individual pens and fitted with ruminal cannulae were grouped (two wethers per group) and fed fresh forage (Italian ryegrass, Lolium multiflorum) with either beet pulp (BP) or corn based concentrate (C) as supplements, using a crossover experimental design. The experiment was carried out during two experimental periods: the growing phase (early spring, G) and the reproductive phase (late spring, R) of the ryegrass. The diet of each animal was changed as the grass moved from the vegetative phase to the reproductive phase. The intake, the chemical composition of the diet and ruminal parameters such as pH, ammonia and volatile fatty acid were determined. Total dry matter intake (DMI) decreased from early to late spring (1081 vs 767 g/d; P

The use of alternative plant protein in place of soybean meal protein in diets for farmed animals aims to reduce the extra-EU soybean import and partially substitute GMO in the food chain. Among the possible alternatives, the heat-processed (flaked) pea appears interesting in dairy cow diets. Two consecutive experiments were carried out to test flaked peas as a partial substitute for soybean meal in the diet of Reggiana breed dairy cows producing milk for Parmigiano-Reggiano cheese-making. In both experiments a “Control” concentrate (8.3% soybean meal) was compared to a “Pea” concentrate (5% soybean meal and 15% flaked peas). Forages fed to animals included mixed grass hay and alfalfa hay in experiment 1, and hay (mixed grass and alfalfa) plus mixed grass in experiment 2. Milk yield and quality, and the characteristics of grab faecal samples, examined to get some empirical indicators of digestibility, were similar between feeding groups. Compositional changes (crude protein and solubility) in forages used as common base in the diets of both experiments had a slight effect on milk and plasma urea contents. There was a tendency for a higher milk urea content in the “Pea” group (32.3 vs 30.1mg/dl in experiment 1, P

The objective of the study was to evaluate the nutritive value of raw, extruded or expanded peas relative to soybean meal in lactating dairy cows feeding. Twenty four Italian Holstein cows (8 primiparous and 16 pluriparous), 604 ± 109 kg body weight, 34.5 ± 2.5 kg/d milk yield, were randomly assigned to four dietary treatments in a 4x4 Latin square arrangement with periods of four weeks and washout period of seven days. Diets were fed ad libitum(5% orts). The bulk of the base diet on a dry matter basis was corn silage (31.2%), alfalfa hay (16.7%), grass hay (4.1%), protein supplement (10.3%), whole cotton seed (8.5%), corn and barley mix (24.9%), soybean meal (3.4%) and calcium soap (0.9%). The pea (2.5 kg/cow/day) partially replaced the soybean meal and totally replaced the barley meal of the base diet. The unprocessed or differently processed pea did not affect the dry matter intake. The extruded pea group had a 3.2% increase (P< 0.05) of the milk yield compared to the control group. When estimated as contrast analysis, the technological treatment (extruded or expanded) on peas did not modify the milk yield and composition. Among pea diets, animal fed the extruded pea had the higher (P< 0.05) milk protein content, although not different than that of the control group. The rumen acetate was reduced (P< 0.05) and the butyrate and valerate were increased (P< 0.05) in animals fed extruded pea compared to the control. No differences were observed among feeding groups on blood parameters except for the cholesterol level higher (P< 0.05) in animals fed the expanded pea diet. There were no effects of diets on milk rennet coagulation characteris- tics. Results support the partial substitution of soybean meal and the total substitution of barley meal with peas in diets for lactating cows with no negative effects on milk yield and composition.

Chemical hazards may occur in any phases of the different livestock production systems. Aim of this review is to address an update about the key issues related to the risk of contamination in foods of animal origin by environmental contaminants linked to industrialisation or urbanisation (e.g., heavy metals and persistent organic pollutants), and natural contaminants (e.g., mycotoxins). This review deals with current issues and future perspectives on the complex issue of the safety of feeds and foods of animal origin, by taking into account the estimation of the occurrence of chemical residues in food, the hazard identification and characterisation of mycotoxins in animal feeds, and the analysis of feedstuffs as a tool to control and evaluate food safety.

The present study investigated in a practical farm condition the response of two commercial maize hybrids (maturity class FAO rating 700) grown for silage production on chemical composition and digestibility of whole maize plant as consequence of a severe water irrigation reduction. Based on different irrigation applications, water restricted (WR) and fully irrigated (FI) plots received 50 and 200 mm of irrigation water, respectively. A split-plot factorial arrangement in a randomised complete block design with two main plots (WR and FI) and two sub-plots (hybrid A and B) with 12 replications/ treatment was performed. Studied parameters were dry matter (DM) yield, harvest index (HI), chemical composition, rumen in situ DM and neutral detergent fibre disappearance (DMD and NDFD, respectively), indigestible NDF (iNDF), 7h in vitro starch degradability (7hIVSD) and net energy (NE) for lactation content. Total DM and grain yields, HI and chemical composition differed (Pvs 56.4% DM) and NDFD (61.0 vs 58.4% NDFOM) were measured for FI with respect to WR crops, whereas iNDF was about 36% higher (P

Thirty multiparous lactating cows were divided in three groups based on milk yield: high (H), average (A) and low (L). Within each group, cows were randomly allotted to two levels of iodine inclusions into the diet and respectively: level 1 (1): base diet containing 1.55 mg/kg dry matter, level 2 (2): base diet plus 47.2 mg/d. Potassium iodide was used. Feeds, orts, drinking water and individual milk samples were collected and analysed for the iodine content. The iodine concentration and total excretion in milk were affected by the level of iodine supplementation (P

A rapid AFB1 detection method by ELISA kit test was used on feedstuff samples, and compared to an HPLC method, to verify if the presence of clay-adsorbent (SA) could cause erroneous quantification of the toxin. Samples were obtained using two AFB1-contaminated feedstuffs (7.92 and 17.58 µg/kg for low and high contaminated feeds; LC and HC respectively), added either one of two commercial SAs (Atox® and Myco AD) and three different inclusion doses (0, 10 and 20 g/kg, respectively for CTR, 1% and 2% doses). The HPLC and ELISA data were compared in CTR samples with a paired t-test. The AFB1 recoveries, performed with ELISA, were analysed as a completely randomized design using a 2×2×3 factorial arrangement. The ELISA method tended to underestimate the AFB1 concentrations with respect to the HPLC method, both in HC (P=0.050) and in LC (P

It was previously reported that injection of anaflatoxin B1 (AnAFB1) conjugated to keyhole limpet hemocyanin (KLH), together with Freund's adjuvant, was effective in inducing in cows a long lasting titer of anti-aflatoxin B1 (AFB1) antibodies (Abs), cross-reacting with other aflatoxins, which were able to hinder, proportionally to their titer, the secretion of aflatoxin M1 (AFM1) into the milk of cows continuously fed with AFB1. According to anti-AFB1 Ab titer, 50% of the vaccinated cows were recognized as high responder animals. In an attempt to prepare a more effective formulation for vaccination of cows, it was compared the immunogenicity, in Holstein Friesian heifers, of AnAFB1 covalently conjugated to KLH or to recombinant diphtheria toxin (CRM197) molecules, and injected together with various adjuvants. This study demonstrated that injection of AnAFB1 conjugated to KLH and mixed with complete (priming) and incomplete Freund's adjuvant (boosters), as in the previous schedule of immunization, was the most effective regimen for inducing Ab responses against AFB1, although pre-calving administration could increase the effectiveness of vaccination, resulting in 100% high responder animals. After one booster dose at the beginning of the milk production cycle, anti-AFB1 Ab titers were comparable to those recorded at the end of the immunization schedule, and proved to be effective in reducing significantly AFB1 carry over, as AFM1, from feed to milk. Pre-calving vaccination of dairy heifers with conjugated AnAFB1, adjuvated with complete and incomplete Freund's adjuvant, may represent the most effective tool for preventing the public health hazard constituted by milk and cheese contaminated with aflatoxins.

The use of alternative plant proteins in place of the soybean meal protein in diets for producing animals aims to reduce the extra-EU soybean import and partially substitute the GMO in the food chain. Among possible alternatives, the heat-processed legume grains seem interesting for dairy cow diets. Two consecutive experiments were carried out to evaluate flaked pea and faba beans as substitute for soybean meal in diets for Reggiana breed dairy cows producing milk for Parmigiano-Reggiano cheese-making. In both experiments a C concentrate (110 g/kg soybean meal, no pea and faba beans) was compared to a PF concentrate (150 g/kg flaked pea, 100 g/kg flaked faba beans, no soybean meal). Forages fed to animals were hay (mixed grass and alfalfa) in experiment 1 and hay plus mixed grass in experiment 2. Concentrate intake, milk yield and milk quality (rennet coagulation traits included) were similar between feeding groups. Parameters on the grab faecal samples, as empirical indicators of digestibility, had a smaller (Pvs 3.1 and 2.3 vs 2.8%, respectively for PF and C in experiment 1 and 2). Some blood indicators of nitrogen metabolism (protein, albumin, urea) were similar between the feeding groups. The inclusion of pea and faba beans, within the allowed limit of the Parmigiano-Reggiano Consortium for diet formulation, could represent a feasible opportunity for a total substitution of soybean meal.

The effect of pea and faba bean inclusion in growing-finishing heavy pig diets was evaluated. The following iso-lysinic and iso-energetic diets offered to the pigs in three phases (40-80; 80-120; 120-160 kg) were compared: CTR, control diet with soybean meal (SBM) as protein source; RP, CTR diet where pea replaced SBM; RF, CTR diet where faba bean replaced SBM. 126 animals were randomly distributed in 3 homogeneous groups with 42 animals each (7 pens with 6 animals each per treatment). The RP and RF diets did not negatively affect the carcass characteristics both of the pigs slaughtered at the conventional weight (127.5 kg) and heavy pigs (158.5 kg). The pigs fed the RP and RF diets ate similarly to the pigs fed the CTR overall the trial but RF pigs grew better than CTR animals. The subcutaneous fat of the fresh hams destined for Parma ham production and obtained from pigs fed RP diet had a higher omega 3 fatty acid percentage. Moreover, the RP and RF diets resulted in fat with better omega 3/omega 6 ratio compared with CTR. All the fat samples had iodine numbers within the limit value (70) reported by the Production Disciplinary of Parma ham. No treatment effect was found on the analytical and sensorial characteristics of the Parma hams, except for the aged taste which was more intense in the hams obtained from pigs fed the RF diet. These results indicate that pea and faba bean may be used as an alternative to imported SBM.

The effect of pea and faba bean inclusion in growing-finishing heavy pig diets was evaluated. The following iso-lysinic and iso-energetic diets offered to the pigs in three phases (40-80; 80-120; 120-160 kg) were compared: CTR, control diet with soybean meal (SBM) as protein source; RP, CTR diet where pea replaced SBM; RF, CTR diet where faba bean replaced SBM. 126 animals were randomly distributed in 3 homogeneous groups with 42 animals each (7 pens with 6 animals each per treatment). The RP and RF diets did not negatively affect the carcass characteristics both of the pigs slaughtered at the conventional weight (127.5 kg) and heavy pigs (158.5 kg). The pigs fed the RP and RF diets ate similarly to the pigs fed the CTR overall the trial but RF pigs grew better than CTR animals. The subcutaneous fat of the fresh hams destined for Parma ham production and obtained from pigs fed RP diet had a higher omega 3 fatty acid percentage. Moreover, the RP and RF diets resulted in fat with better omega 3/omega 6 ratio compared with CTR. All the fat samples had iodine numbers within the limit value (70) reported by the Production Disciplinary of Parma ham. No treatment effect was found on the analytical and sensorial characteristics of the Parma hams, except for the aged taste which was more intense in the hams obtained from pigs fed the RF diet. These results indicate that pea and faba bean may be used as an alternative to imported SBM.

Hydrophobic prolamins are endosperm storage proteins accounting for about 40% of the total protein in most cereals seeds. Despite the absence of a reference method, several procedures have been periodically published to quantify prolamins in cereals. The aim of this study was to compare a conventional fractionation assay (LND) vs three other methods: one based on sequential extractions (HAM) and two rapid turbidimetric procedures (L&H and DRO). Prolamins were extracted in duplicate on barley, corn and wheat samples. For the turbidimetric prolamin evaluation in barley and wheat, a universally available purified gliadin, as alternative to purified zein, was also tested as standard reference material (SRM). The extraction prolamin values were different among grain types (P0.05). LND agreed sufficiently well both with HAM and with L&H methods (R2=0.664 and R2=0.703, respectively, P0.05), whereas a higher prolamin quantification was obtained using HAM (P

The potential of anaflatoxin B(1) (AnAFB(1)) conjugated to keyhole limpet hemocyanin (KLH) as a vaccine (AnAFB(1)-KLH) in controlling the carry over of the aflatoxin B(1) (AFB(1)) metabolite aflatoxin M(1) (AFM(1)) in cow milk is reported. AFB(1) is the most carcinogenic compound in food and foodstuffs amongst aflatoxins (AFs). AnAFB(1) is AFB(1) chemically modified as AFB(1)-1(O-carboxymethyl) oxime. In comparison to AFB(1), AnAFB(1) has proven to be non-toxic in vitro to human hepatocarcinoma cells and non mutagenic to Salmonella typhimurium strains. AnAFB(1)-KLH was used for immunization of cows proving to induce a long lasting titer of anti-AFB(1) IgG antibodies (Abs) which were cross reactive with AFB(1), AFG(1), and AFG(2). The elicited anti-AFB(1) Abs were able to hinder the secretion of AFM(1) into the milk of cows continuously fed with AFB(1). Vaccination of lactating animals with conjugated AnAFB(1) may represent a solution to the public hazard constituted by milk and cheese contaminated with AFs.

Hydrophobic prolamins are endosperm storage proteins accounting for about 40% of the total protein in most cereals seeds. Despite the absence of a reference method, several procedures have been periodically published to quantify prolamins in cereals. The aim of this study was to compare a conventional fractionation assay (LND) vs three other methods: one based on sequential extractions (HAM) and two rapid turbidimetric procedures (L&H and DRO). Prolamins were extracted in duplicate on barley, corn and wheat samples. For the turbidimetric prolamin evaluation in barley and wheat, a universally available purified gliadin, as alternative to purified zein, was also tested as standard reference material (SRM). The extraction prolamin values were different among grain types (P0.05). LND agreed sufficiently well both with HAM and with L&H methods (R2=0.664 and R2=0.703, respectively, P0.05), whereas a higher prolamin quantification was obtained using HAM (P

Eight potential aflatoxin-sequestering agents (SAs) were tested for their ability to adsorb aflatoxin B1 (AfB1) and aflatoxin G1 (AfG1) in vitro. They belong to main SA classes: silicate minerals (calcium, magnesium and sodium bentonites, kaolinite, zeolite and clinoptinolite), activated carbon and yeast cell wall-derived. The AfB1 and AfG1 used in present work were extracted from a contaminated corn meal (82.21 mg/kg of AfB1 and 97.20 mg/kg of AfG11). Three single-concentration adsorption tests, consisting of a simply-water (W), a gastrointestinal simulating monogastric model (MM) and a ruminant model (RM), were used. The methods differed for dilution media, incubation steps and pH condition in which they were conducted. In particular, one step (2h at 39°C) at pH 7 for W; two steps (4h at 39°C) at pH 2 and 7 for MM; and a pre-incubation in rumen fluid (pH 7 for 2h at 39°C) + two steps (4h at 39°C) at pH 2 and 7 for RM, characterized each method. The AfB1:SA ratio (g/g) and dilution factor (ng of incubated AfB1:mL of volume) were chosen (1:500,000 and 4.1, respectively) to reflect field conditions. The AfB1 and AfG1 recovered in controls were 92.3% and 104.9% in W and 89.5% and 101.5% in MM; while in RM were 65.2% and 81.9%; respectively. This supported the idea of intrinsic rumen fluid factors could be involved in sequestering of aflatoxins. In the present study, three SAs (activated carbon, Mg bentonite and Na bentonite) were very efficient to sequester the available AfB1, with a sequestering activity of over 99.0% with each method. The Ca bentonite and clinoptinolite were able to bind available AfB1 in MM and RM methods, while they appeared inefficient (available AfB1 sequestered less than 80%) when W was used. The adsorption ability of zeolite was confirmed only with the W method. Ineffective or limited sequestering activity were obtained with kaolinite and yeast cell wall-derived products with each method. The AfB1 and AfG1 sequestering efficiencies observed in the present work resulted very similar showing strong and positive correlation (P in vitro pre-screening of potential sequestering agents. However, the major practical and analytical implications related to rumen fluid method suggested that MM method should be used.

The use of alternative plant proteins in place of the soybean meal protein in diets for farmed animals aims to reduce the extra-EU soybean import and partially substitute the GMO in the food chain. Among the possible alternatives, the heat-processed (flaked) faba beans appears interesting for dairy cow diet. Two consecutive experiments were carried out to test flaked faba beans as a partial substitute for soybean meal in the diet of Reggiana breed dairy cows producing milk for Parmigiano-Reggiano cheese-making. In both experiments a “Control” concentrate (12% soybean meal, no faba beans) was compared with a “Faba” concentrate (7.5% soybean meal and 10% flaked faba beans). Forages fed to animals were hay (mixed grass and alfalfa) plus mixed grass in experiment 1, hay only in experiment 2. Milk yield and quality and the characteristics of grab faecal samples as empirical indicators of digestibility, were similar between feeding groups. The milk urea content was slightly lower in the “Faba” group, particularly in experiment 2 (“Control” vs “Faba”: 34.6 vs 32.9 mg/dL in experiment 1, P

Six pregnant heifers, coming from a herd with a history of high concentration of polychlorobifenyls (PCB) into the milk, were fed with a very low-PCB diet starting from the 6th month of pregnancy. After parturition cows were milked for at least 190 d with a maximum of 270 d. Diet was made of: corn silage (31.7% of DMI), dehydrated alfalfa (13.3% of DMI), grass hay (3.7% of DMI) and concentrate (51.3% of DMI). The average DMI was 23.12 kg/d. Milk production was recorded and samples of milk and blood were taken and analyzed for PCB (18, 28, 31, 52, 44, 101, 149, 118, 153, 138, 180 and 194 congeners) content using a GC-MS technique. The average milk yield (kg/d) of the 6 cows was 26.0, 22.5, 23.2, 24.5, 28.9, 29.3. The maximum PCB concentration of 100 ng/g of fat allowed by the Italian law was reached after 144-209 d of lactation. One animal after 204 days had a PCB concentration of 102 ng/g fat. If log10 of PCB concentration (ppb) in milk fat is regressed against days in milking (DIM) the following significant equation was obtained: log10 PCB (ng/g of milk fat)= 2.796 -0.00474 DIM; r2 0.72; P