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The quantification of feed composition parameters, as well as feed digestibility, requires time-consuming and expensive laboratory reference methodologies. In the context of precision livestock feeding, fast, accurate and on-line methods to quantify at a cost-effective the composition and characteristics of total mixed ration (TMR) are desired tools. Therefore, the aim of the present study was to develop prediction models using near infrared spectroscopy (NIRS) for a plethora of TMR compositional traits and digestibility. A total of 205 TMR samples were collected in herds located in the Parmigiano Reggiano area in different experimental trials. Samples were analysed, using reference methodologies, for crude protein (CP), starch, amylase-treated ash-corrected neutral detergent fiber with addition of sodium sulfite (aNDFom), acid detergent fiber (ADF), acid detergent lignin (ADL) and ash. Moreover, the fiber undegradable fractions (uNDF) of TMR was quantified in vitro at different timepoints (24 h, 30 h, 120 h, 240 h). Spectral data in the range from 900 to 2500 nm were collected for all these samples using a TANGO FT-NIR spectrometer. Partial least squares regression was employed to calibrate NIRS prediction models through a cross-validation, but also to validate such models on a subset including a random 30% of the total observations which were excluded from the calibration set. Different mathematical pre-treatments were also applied to the spectra data in order to identify the transformation which provided the most accurate NIRS prediction model (in terms of maximised explained variance and minimised root mean square error of prediction). The coefficient of determination in external validation (R2P) was >0.80 only for starch, between 0.60 and 0.80 for CP, aNDFom, ADF and ADL, while

Focus of this study was to evaluate the most suitable breed/crossbred and the appropriate nutritional strategies to increase marbling in beef muscle and to improve its healthy properties, in particular the n-3 fatty acids content. One hundred and seventy-six heifers of three crossbreed commonly reared by Emilia-Romagna farmers: 48 Bleu Belge × Freisian (ITA), 48 Charolais × Aubrac (FRA), and 80 Angus (ANG) were used. Animals of each breed were randomly subdivided in two experimental groups that received two diets, differing for the dietary lipid source. Control group (C) received a basal diet containing protected vegetable fats, whereas treated one (T) received the same basal diet supplemented with 0.9 kg/head/day of extruded flaxseed. After slaughtering, a sample of Longissimus thoracis was collected from each animal for meat quality analysis. Our results demonstrated that, in a shorter fattening period, ANG animals obtained the best dry matter intake, average daily gain and the best fattening scores. ITA and ANG meat presented the highest marbling scores. ANG breed had the highest amount of C18:1, the highest unsaturated/saturated fatty acids ratio and the lowest n-6/n-3 ratio. The T animals, independently from breed, showed the highest amount of α-linolenic acid (ALA). In addition, ANG-T meat presented the highest ALA content.

In vitro and in situ procedures performed to estimate indigestible neutral detergent fiber (iNDF) in forage or fecal samples are time consuming, costly, and limited by intrinsic factors. In contrast, near infrared reflectance spectroscopy (NIRS) has become widely recognized as a valuable tool for accurately determining chemical composition and digestibility parameters of forages. The aim of this study was to build NIRS calibrations and equations for fecal iNDF. In total, 1,281 fecal samples were collected to build a calibration data set, but only 301 were used to develop equations. Once dried, samples were ground and chemically analyzed for crude protein, ash, amylase and sodium sulfite-treated NDF corrected for ash residue (aNDFom), acid detergent fiber, acid detergent lignin, and in vitro digestion at 240 h to estimate iNDF (uNDF240). Each fecal sample was scanned using a NIRSystem 6500 instrument (Perstorp Analytical Inc., Silver Spring, MD). Spectra selection was performed, resulting in 301 sample spectra used to develop regression equations with good accuracy and low standard error of prediction. The standard error of calibration (SEC), cross validation (SECV), and coefficients of determination for calibration (R2) and for cross validation (1 - VR, where VR = variance ratio) were used to evaluate calibration and validation results. Moreover, the ratio performance deviation (RPD) and ratio of the range of the original data to SECV (range/SECV; range error ratio, RER) were also used to evaluate calibration and equation performance. Calibration data obtained on fiber fractions aNDFom (R2 = 0.92, 1 - VR = 0.87, SEC = 1.48, SECV = 1.89, RPD = 2.80, and RER = 20.19), uNDF240 (R2 = 0.92, 1 - VR = 0.86, SEC = 1.65, SECV = 2.24, RPD = 2.57, and RER = 14.30), and in vitro rumen aNDFom digestibility at 240 h (R2 = 0.90, 1 - VR = 0.85, SEC = 2.68, SECV = 3.43, RPD = 2.53, and RER = 14.0) indicated the predictive equations had good predictive value.

The objective of this study was to investigate the effects of 2 alfalfa hays differing in undigested neutral detergent fiber content and digestibility used as the main forage source in diets fed to high producing cows for Parmigiano-Reggiano cheese production. Diets were designed to have 2 different amounts of undigestible NDF [high (Hu) and low (Lu)], as determined by 240-h in vitro analysis (uNDF240). Alfalfa hay in vitro digestibility [% of amylase- and sodium sulfite-treated NDF with ash correction (aNDFom)] at 24 and 240 h was 40.2 and 31.2% and 53.6 and 45.7% for low- (LD) and high-digestibility (HD) hays, respectively. The 4 experimental diets (Hu-HD, Lu-HD, Hu-LD, and Lu-LD) contained 46.8, 36.8, 38.8, and 30.1% of alfalfa hay, respectively, 8.6% wheat straw, and 35.3% corn (50% flake and 50% meal; DM basis). Soy hulls and soybean meal were used to replace hay to balance protein and energy among diets. Eight multiparous Holstein cows (average milk production = 46.0 ± 5.2 kg/d, 101 ± 38 d in milk, and 662 ± 42 kg of average body weight) were assigned to a 4 × 4 Latin square design, with 2 wk of adaptation and a 1-wk collection period. Dry matter and water intake, rumination time, ruminal pH, and milk production and composition were measured. Diets and feces were analyzed for NDF on an organic matter basis (aNDFom), acid detergent fiber, acid detergent lignin, and uNDF240 to estimate total-tract fiber digestibility. Dry matter intake and rumination times were higher in HD diets compared with LD diets, regardless of forage amount. Rumination time was constant per unit of dry matter intake but differed when expressed as a function of uNDF240, aNDFom, or physically effective NDF intake. No differences were found among treatments on average ruminal pH, but the amount of time with pH

Rumination and activity behaviours are important welfare indexes in beef-cattle housing. The main objective of the present study was to assess whether the automatic collars are able to reliably assess the rumination and activity patterns of beef heifers. For this purpose, individual rumination time and activity (RT and AT respectively) were continuously recorded using an automatic neck-collar system (Hr-Tag, SCR Engineers Ltd, Israel) on the three trials performed. For Experiment 1, 60 Italian crossbred heifers were randomly assigned to one of two experimental diets for 9 months: the corn–silage diet (CS), which included 50% forage on a dry-matter (DM) basis (43% corn–silage, 7% wheat straw) and the hay diet (HAY), with 57% forage on DM basis (28.5% grass hay, 28.5% alfalfa hay). Heifers consuming HAY diet showed greater (P < 0.05) RT (min/day) and AT (bits/day) than did those on CS diet. Rumination time per kilogram of dry matter and per kilogram of amylase- and sodium sulfite-treated neutral detergent fibre corrected for ash residue (aNDFom) intake were similar in the two experimental groups, while RT per kilogram of physical effective NDF (peNDF) intake was greater (P < 0.05) in the CS group than in the HAY one. Daily rumination and activity patterns (min/2 h and bits/2 h respectively) were significantly different between CS and HAY groups. In the second experiment, 32 beef heifers were randomly allocated in two homogeneous pens, each containing eight animals, and two non-homogeneous ones, in which animals were added at different periods. The AT of non-homogeneous pens was significantly higher than that of the homogeneous ones, suggesting a distress condition for values higher than 309 bits/day AT. In the third experiment, RT and AT of three animals with respiratory disease were collected using the automatic collar system and compared with AT and RT of the healthy animals. Sick animals presented a significantly lower RT and higher AT than did the healthy ones. The cut-off to distinguish sick from healthy heifers was set to 537 bits/day AT and 381 min/day RT. In conclusion, the present study demonstrated that automatic collars can reliably monitor different rumination and activity behaviours of beef animals in various management conditions and different health statuses.

The aim of this study was to develop near-infrared reflectance spectroscopy (NIRS) calibrations for estimation of chemical components (protein, fat, ash and moisture) and the principal fatty acid (FA) groups (SFA, MUFA, PUFA, n−3, n−6) in beef meat. Beef samples (n = 207) of three different muscles (Longissimus dorsi, Rectus abdominis and Semitendinosus) were used for the study. The calibration models were performed both on raw and freeze-dried (FD) meat. Raw meat predictions for protein, fat and ash were performed, using reference values expressed as percentage on dry-matter (DM) or fresh-minced (FM) weights. In addition, FD meat predictions for the same chemical components were developed. FA predictive models were based on the relative composition of FA (weight percent of total FA). The predictability of calibrations based on FD meat was higher than that of models based on raw samples, except for n−3. Regarding the raw meat, the calibration models obtained from reference values expressed on DM basis were better than those obtained from data expressed on FM basis. In addition, because of the low coefficients of determination of prediction (R2p) and the low RPD obtained, NIRS models of FA families are not useful for a quantitative fatty acid analysis.

Long-term in vitro fermentation (240 h) evaluating amylase-treated, ash-corrected neutral detergent fiber, with addition of sodium sulfite (aNDFom) digestibility is required to quantify the indigestible fiber fraction. It is commonly accepted to inoculate rumen fluid more than one time during such fermentations, every 96 h or at 120 h. However, no studies have been conducted to verify if the reinoculation is actually required to properly carry out the fermentation process. The current study aims to evaluate the effects of these procedures on aNDFom digestibility at 240 h. The study was conducted on a total of 24 forage samples (8 alfalfa hays, 8 grass hays, and 8 corn silages). Samples were digested in triplicate at 240 h in vitro. Rumen fluid was added twice (at 96 and 192 h) in treatment 1, after 120 h in treatment 2, whereas no addition was made in treatment 3. At the end of the fermentations, residual aNDFom was quantified to calculate digestibility. Among treatments, no difference was found in digestibility of aNDFom. Moreover, treatment 1 resulted in higher variability compared with other treatments. Results obtained in the current study show that subsequent addition of rumen fluid is not necessary for a proper estimation of aNDFom digestibility, and can be avoided.

We investigated the effects of monensin controlled-release capsule (CRC; Kexxtone, Eli Lilly and Company Ltd., Indianapolis, IN) preventative ketosis treatment on the traditional cheesemaking process as well as the final characteristics of Parmigiano Reggiano (PR) cheese. The use of this prevention product to reduce the incidence of ketosis in transition dairy cows was approved by the European Medicines Agency in 2013. No previous studies are available concerning the effects of this treatment on prolonged-ripening cheese produc-tion such as PR. In PR cheese production, feed, feed additives, and cow treatments are strictly regulated to avoid any possible interference with traditional manufacturing processes. For these reasons, on 1 farm where all milk was used for PR cheese production, monensin CRC was administered to 33 cows, 21 d before calving in the monensin-treated group (TRT), whereas untreated cows with similar breed and parity characteristics constituted the control group (CTR). For 20 wk, milk obtained from each group and whey starter were separately managed and transported in the cheese factory, where 2 cheese wheels per group were produced daily, making 552 PR cheese wheels in total. Morning bulk tank milk composition, cheesemaking properties, and whey starter fermentation activities were analyzed twice a week. Every aspect of the cheesemaking process was recorded and the resulting cheese was evaluated after 36 h and 6, 12, and 18 mo from production for yield, texture defects, composition, and fatty acids profile. Milk from the 2 groups differed for somatic cell content (TRT = 3.04 vs. CTR = 4.06, somatic cell score), total bacterial count (TRT = 4.08 vs. CTR = 6.08 × 1,000 cfu/mL), titratable acidity (TRT = 3.66 vs. CTR = 3.72 Soxhlet-Henkel degrees/50 mL), and casein content percentage (TRT = 2.4 vs. CTR = 2.5%). Whey starter parameters were comparable between the 2 groups. Final cheese composition and organoleptic profile were not influenced by the treatment, except for C18:1 content being enhanced (TRT = 22.8 vs. CTR = 20.8% of fatty acids). Percentage of defected ripened cheese was significantly lower in the treated group, both at x-ray evaluation performed at 6 mo (TRT = 6.2 vs. CTR = 12.3%) and at the consortium inspection, performed at 12 mo of ripening (TRT = 1.5 vs. CTR = 6.5%). On the other hand, average cheese yield at 18 mo of ripening was partially reduced (TRT = 7.5 vs. CTR = 7.7%). Overall, the use of mo-nensin CRC had no negative effect on the cheesemaking process, prolonged ripening cheese characteristics, milk composition, or whey starter quality.

The intramuscular fat (IMF), recognized by the consumer as marbling, is an important meat quality trait. The objective of this study was to validate a new method of beef marbling evaluation by image analysis. The new assessment was compared with two known marbling measurements: chemical IMF and USDA scores. Moreover, the relationship between marbling measurements and other meat quality parameters was investigated. Samples of Longissimus thoracis et lumborum (LTL) muscle were obtained from carcasses of 39 Italian crossbred heifers and 62 Angus bred heifers, that underwent two different dietary treatments. The use of various breeds and diets was chosen to obtain different intramuscular fat levels, in order to validate the use of Image J software for the analysis of different type of beef meat. The images of fresh cuts were appraised by experienced beef graders, and the samples were used to determine fat content by chemical Soxhlet extraction. Carcasses measurements according to the EUROP system, and other physical meat proprieties were also assessed. The results demonstrated that the marbling measurements obtained by computer image analysis, such as the number of marbling particles, the average particle size (mm), and the percentage of marbling particles (%), significantly (P < 0.05) correlated with USDA scores and IMF content. Moreover, the principal component analysis (PCA) showed three principal meat components, identified as 1) color, 2) fat, and 3) water release. The second principal component (PC) explained 24.94% of variance, and was positively correlated with image analysis measures, USDA score, and IMF, while negatively correlated with the Warner–Bratzler shear force (WBSF).

Gossypol is a yellow pigment occurring in all parts of cotton plants, with the highest levels found in seeds, and it exhibits a variety of toxic effects. Few data are available on the content of gossypol in the commercial complementary feed and in feed raw materials. The present study was focused on the investigation of the presence of free gossypol in commercial complementary feed not containing cotton. A total of 50 samples of commercial complementary feed for dairy cows were performed in 29 feed mills both using and not using cotton as feed material. The free gossypol contamination resulted under the detection limit of the technique (4 mg/kg) in 12 out of 50 samples analysed and ranged from 4 to 20 mg/kg in 28 samples. In 10 samples the level of free gossypol ranged from 20 to 29.5 mg/kg. Average contamination of samples was 12.2±9.2 SD mg/kg. No significant difference (P=0.571) was shown in free gossypol concentration between feed produced in cotton free plants and in plants where cotton is used as feed material. Free gossypol content detected in the present study allows considering complementary feed for dairy cows not at risk. On the other hand, the detection of free gossypol in cotton free complementary feed, probably attributable to cross contamination of feed materials upstream of the feed mill, should be further investigated.

In vitro methods requiring ruminal microorganisms to ferment and digest feeds have been used for decades. Though commonly accepted, collecting and pooling rumen fluid from different donor animals to avoid individual characteristics could affect in vitro fermentations. The current study evaluated the effects of individual or pooled liquors on in vitro digestibility of amylase- and sodium sulfite-treated NDF with ash correction (aNDFom). The study was conducted on 24 samples (8 alfalfa hays, 8 grass hays, and 8 corn silages). The 3 donor animals (treatment 1, 2, and 3) were selected based on similar body weights, parity, days in milk, milk production, and milk composition. Samples were digested in vitro via inoculation of different rumen fluid at different time points (12, 24, 72, and 120h). An equal amount of each liquor collected was sampled and equally mixed with the others to obtain treatment 4. For the alfalfa hay group, differences were observed at 12 (29.95, 27.07, 29.02, and 32.55% aNDFom for treatments 1, 2, 3, and 4, respectively) and 24h (37.35, 35.54, 36.44, and 40.56% aNDFom for treatments 1, 2, 3, and 4, respectively). The inoculum source did not affect in vitro digestibility over longer time periods (72 and 120 h). Similar results were observed in the grass hay group, in which the mixed inoculum had greater digestibility values at both 12 (28.86, 26.89, 27.88, and 30.92% aNDFom for treatments 1, 2, 3, and 4, respectively) and 24h (37.35, 35.54, 36.44, and 40.56% aNDFom for treatment 1, 2, 3, and 4, respectively), but not over longer time periods. For the corn silage group, we observed differences for treatment 4 only at 12h (35.78, 33.87, 34.83, and 37.80% aNDFom for treatment 1, 2, 3, and 4, respectively). These results underline the differences among donor animals, especially when evaluating short incubation time points, and that pooling rumen contents is not equal to averaging across individual animals. Reported data require a deeper investigation on whether or not the method of inoculating a pool of rumen contents represents the actual ability of the animal to digest fiber.

This study investigated aflatoxin B1 (AFB1) contamination in dairy cow feed and the risk management of AFB1 content in concentrates undertaken by feed industries in the Parmigiano Reggiano area. Data on aflatoxin contamination risk management applied in 29 feed industries were collected and the AFB1 content of 70 feed samples was analysed. Data were collected within the framework of a quality control programme promoted by the Parmigiano Reggiano Consortium in 2013 and 2014. Audit results showed that the control procedures to prevent AFB1 contamination mainly focused on maize and its by-products. AFB1 concentration resulted lower than 5 ppb [legal European Union (EU) limit] in all samples; in one out of 70 samples, AFB1 content was 3.8 ppb and in all the other samples it was lower than 3 ppb. Results showed that AFB1 risk management applied by Italian feed industries effectively monitors AFB1 levels in feed below the EU legal limit.

Forage neutral detergent fiber (NDF) content and particle size are important factors that affect rumen function. The aim of the current study was to evaluate the effects on rumen health, NDF digestibility, and animal performance of pelleting a forage-based diet. Eight Holstein heifers (age 336 ± 30 d, body weight 346 ± 35 kg) were randomly assigned to a repeated crossover design. Animals were housed in tie-stalls and fed for ad libitum intake. The study included 4 periods of 3 wk, the first 2 wk for adaptation to the diet and the last wk for data collection. Diets had the same ingredients but had a different physical form: total mixed ration (TMR) and pellet (diameter = 8 mm). The physically effective NDF (peNDF) differed between the 2 treatments (39.8 and 11.8% of NDF in the TMR and pellet diets, respectively). During the trial, dry matter intake (DMI), water intake, rumination time, rumen temperature, and pH were evaluated daily. Fecal samples were collected in wk 3 of each period to determine total-tract digestibility of the potential digestible (pd)NDF. Average daily gain and feed conversion ratio were calculated at the end of each period. With the pellet diet, DMI, DMI/body weight, and water consumption were higher. We observed no significant difference in average daily gain or feed conversion ratio. Rumination time was lower for the pellet diet than for the TMR diet (241 vs. 507 min/d, respectively). Diet had no effect on rumen temperature or rumen pH. The total-tract digestibility of the pdNDF was greater with the TMR diet than with the pellet diet (90.25 vs. 86.82% pdNDF, respectively). The results of the current study suggest that a complete-feed pellet diet was well accepted by the animals, as demonstrated by higher DMI. Rumination time was reduced with the pellet diet, but rumen pH was not different. The pdNDF digestibility was high for both diets, but significantly higher for the TMR diet. Given that animal performance was similar between the 2 diets, although they differed with respect to DMI and fiber digestion, we hypothesize that the 2 diets had different retention times, related to their physical form. A complete-feed pellet diet formulated to provide a sufficient level of NDF from forages could be fed to growing ruminants without apparent negative effects on rumen health and animal productivity, at least for a short period. More research over a longer growing period is needed before recommending this feeding strategy for growing heifer

This study focused on changes in fibrous and protein fractions, changes in fiber digestibility and amount of indigestible neutral detergent fiber (NDF) as a consequence of increased maturity in alfalfa. A total area of 720 m2 was divided in 18 blocks randomly assigned to 3 treatments, differing in cutting intervals. Treatment 1 was harvested with a 21-d cutting schedule, at a prebloom stage; treatment 2 with a 28-d schedule, at about first-bloom stage; whereas a full bloom was observed in treatment 3, harvested with a 35-d cutting schedule. Treatments were replicated 4 times through the springsummer period for 2 subsequent years, 2011 and 2012. Statistical differences were observed for crude protein [treatment 1: 20.8%, treatment 2: 17.3%, and treatment 3: 17.0%; standard error of the mean (SEM) = 0.83], soluble protein, and nonprotein nitrogen among treatments on a dry matter basis. Similar results were observed for acid detergent lignin (6.3, 6.9, and 7.3%, respectively; SEM = 0.39), lower in treatment 1 compared with others, and in vitro NDF digestibility at 24 or 240 h. Indigestible NDF at 240 h resulted in lower values for treatment 1 compared with treatments 2 and 3 (15.5, 17.2, and 18.3%, respectively; SEM = 1.54). Moreover, the indigestible NDF:acid detergent lignin ratio varied numerically but not statistically among treatments, being as much as 9% greater than the 2.4 fixed value applied for rate of digestion calculation and Cornell Net Carbohydrate Protein System (Cornell University, Ithaca, NY)-based model equations. Assuming the diet composition remained unchanged, treatment 3 (35-d cutting interval) would be expected to yield 1.4 kg less milk per day based on energy supply, and 2.8 kg less milk daily based on protein supply than treatment 1.

Myostatin or growth and differentiation factor 8 (MSTN or GDF8, respectively) is a member of the transforming growth factor-β superfamily that acts as a negative regulator of skeletal muscle development and growth. The myostatin encoding gene (MSTN) and the 5’ regulative region in different species has been investigated. The MSTN gene consists of three exons and two intronic regions. In cattle, the gene has well-characterized mutations determining phenotypes with increased muscle mass. In a previous study, by alignment of nucleotide sequences of PCR products of horse MSTN gene, we identified some SNPs of which two T>C transitions in the promoter of the gene. The aim of this work is to study in different horse breeds the frequency of the polymorphism located 516 nt upstream of the ATG start codon. This SNP is within a TATA box sequence (NATAAAA, where N= T or C) and the comparative sequence analysis of mammals myostatin gene promoters reveals that this TATA-box motive is conserved in position and sequence among some species including cattle, sheep and goat. The point mutation is not recognised by restriction enzymes, then the genotyping was done by restriction site insertion-PCR. The primer pair was designed on the basis of the obtained nucleotide sequences. The reverse primer was modified and designed with a single nucleotide mismatched with respect to target sequence to introduces an SspI artificial restriction site when the T nucleotide occurred. The PCR products of 203 bp were digested and the resulting fragments were resolved on 3.5% agarose gel. Two hundred and six samples of DNA belonging to 18 horse breeds (Bardigiano, 20; Breton, 5; Criollo, 10; Esperia Pony, 4; Haflinger, 9; Italian Heavy Draught Horse, 24; Italian Saddle, 21; Italian Trotter, 16; Lipizzan, 11; Maremmano, 13; Murgese, 12; Norico, 10; Purebred Spanish Horse, 10; Salernitano, 12; San Fratellano, 3; Tolfetano, 7; Thoroughbred, 11 and Ventasso Horse, 8) were genotyped. The T allele occurs in all the breeds. The C allele, which eliminates TATA box motif, was detected in 11 breeds and its value frequency ranged from 0.006 (Thoroughbred) to 0.400 (Norico). The homozygous C/C genotype was detected in Haflinger, Bardigiano, Norico and Tolfetano breeds. The presence of the polymorphism within a conserved region of promoter of horse myostatin gene could affect expression gene. This marker could be used for association studies with performance traits in horses.