Management of dairy heifers: Can operant conditioning be an effective and feasible tool to decrease stress and ease animals' close contact and handling?

Besides health monitoring, a regular check of dairy heifers' growth rate is desirable, but it is rarely done because procedures that require restraint and handling can be associated with substantial stress for both animals and farmers. Inexperienced heifers, especially if they are highly responsive to humans, may find restraint and handling potentially aversive. This study investigated whether training heifers of different age and responsiveness toward humans (RTH), through operant conditioning, could reduce stress in animals, ease close contact and handling, and be feasible in terms of farmers' effort. We assessed 60 Holstein heifers of 2 age classes (young, n = 29, 291 ± 39 d; old, n = 31, 346 ± 62 d) according to the avoidance distance test and classified them as confident (n = 20), neutral (n = 21), or nonconfident (n = 19). Half of the heifers of each age and RTH class were trained (n = 29), whereas the other half was not (n = 31). The trained heifers were subjected to target training for 8 sessions and positively reinforced with feed to allow being touched on the muzzle, rump, and perineum. If a heifer refused positive reinforcement, the trainer stepped back as negative reinforcement. In the last week of the experiment, the effect of training on the reaction to handling was assessed in all heifers. We measured heart rate, root mean square of successive interbeat interval differences (RMSSD), and fecal cortisol metabolites (FCMet). The presence of behavioral distress signs was recorded as well. The avoidance distance test was performed a second time 24 h after the measuring session. All of the trained heifers, regardless of RTH class, successfully accomplished the target training task in 6 sessions, each spending on average 25.3 s per session. All of the trained heifers allowed touches on the rump and perineum at the end of the fourth session. Training nonconfident heifers required more time compared with the others. Trained heifers showed higher RMSSD than nontrained heifers (14.2 vs. 16.9 ms, respectively), indicating a lower vagal tone, and thus, a slightly lower stress level than nontrained heifers. Training did not lead to differences in HR, FCMet, or presence of stress behavioral signs. Nonconfident heifers had the highest mean baseline FCMet values compared with neutral and confident heifers (38.4 vs. 30.3 vs. 29.1 ng/g, respectively). Nonconfident heifers also showed the lowest value of FCMet 12 h after the measuring session (36.7 vs. 44.6 vs. 49.7 ng/g), likely due to a decreased responsiveness of the adrenal gland to a stressor. The average avoidance distance decreased between the beginning and the end of the experiment, especially for neutral and nonconfident heifers, regardless of whether they were trained or not. These results show how using operant conditioning on some heifers not only decreased their vagal tone, but also reduced the responsiveness to humans of all the animals, trained and not trained; in the latter case, this reduction was through nonassociative learning, such as habituation.

Effect of feeding fine maize particles on the reticular pH, milk yield and composition of dairy cows.

The particle size of cereal grains has been found to modulate the rate of passage from the rumen and the digestibility of starch and neutral detergent fibre (NDF), but few studies have examined its impact on reticular pH. The study aimed to evaluate the effect of feeding finely ground maize on the risk of ruminal acidosis, milk yield and composition. Twelve Holstein-Friesian cows were assigned to one of two experimental groups and fed according to a cross-over design. Diets were isoenergetic and isonitrogenous and were characterised by the same NDF and ADF, differing only in maize particle size. In the control diet (Ct), the maize meal was ground to 1.0 mm, whereas in the experimental diet, it was finely ground (Fg) to 0.5 mm. The pH and temperature of the reticulum were continuously measured in eight cows throughout the trial using indwelling sensors. Dry matter intake was higher in cows offered Fg diet than in Ct (19.0 vs. 20.3 kg/day; p = 0.067). However, milk yield (p = 0.855) and the 3.5% fat-corrected milk (FCM) (p = 0.724) did not show any differences between the diets. Casein (2.48 vs. 2.57%; p = 0.035) and crude protein (CP) (3.18 vs. 3.31%; p = 0.021) resulted higher in Fg. Similarly, starch digestibility increased in animals offered Fg diet versus Ct (0.94 vs. 0.98; p = 0.078). Among the reticular parameters, the Fg-fed cows spent a significantly higher time below the 5.5 pH threshold (15 vs. 61 min/day; p = 0.047) and had an average daily variation in reticular pH characterised by a lower nadir pH (5.95 vs. 5.72; p < 0.001) and a higher pH range (0.79 vs. 0.94; p = 0.003). In this study, grain particle size affected the risk of the onset of ruminal acidosis. Therefore, it should be carefully considered when formulating rations.

Identification of undeclared sources of animal origin in canine dry foods used in dietary elimination trials.

Failure to respond to commercial limited antigen diets can occur in dogs kept on a dietary trial for the diagnosis of adverse food reaction (AFR). The aim of this study was to assess twelve canine dry limited antigen diets (eleven novel protein diets and one hydrolysed diet) for potential contamination by ingredients of animal origin not mentioned on the label. The validity of the two methods adopted for the detection of such food antigens was also evaluated. Each dietary product was analysed by microscopy analysis using the official method described in Commission Regulation EC 152/2009 with the aim of identifying bone fragments of different zoological classes (mammalian, avian and fish) and by polymerase chain reaction (PCR) for the identification of DNA of animal origin. Discrepancies between the results obtained by PCR and/or microscopy analysis and the ingredients listed on pet food packages were found. Only in two pet foods did the results of both analyses match the ingredients listed on the label. In the remaining ten samples, microscopy detected bone fragments from one or two unpredicted zoological classes, revealing avian fragments in six of ten samples followed by those of fish in five of ten and mammalian fragments in four of ten. In two samples, microscopy analysis identified a contamination that would have otherwise passed unobserved if only PCR had been used. However, PCR confirmed the presence of all the zoological classes detected by microscopy and also identified the DNA of an additional unexpected zoological class in two samples. Dogs might fail to respond to commercial limited antigen diets because such diets are contaminated with potential allergens. Both PCR and microscopy analysis are required to guarantee the absence of undeclared animal sources in pet foods. Before ruling out AFR, a novel protein home-made diet should be considered if the dog is unresponsive to a commercial regimen.

Evaluation of indirect biomarkers for detecting corticosteroids used as illegal growth promoters in beef cattle.

The histological status of the thymus, blood cortisol concentration and circulating neutrophil:lymphocyte ratio were evaluated in 349 slaughtered beef cattle, to assess the potential of these parameters as indirect biomarkers of the illegal use of corticosteroids in meat production. The livers of 20 of the animals were analysed chemically for residues of corticosteroids. The morphology of the thymus was examined for adipose tissue infiltration, cortical atrophy and 'starry sky' appearance, and on the basis of these characteristics, the animals were considered to be negative, suspected or positive for illegal corticosteroid treatment. The animals considered to be negative had a mean cortisol concentration that was significantly higher (29 ng/ml) than that of the animals suspected for corticosteroid treatment (22 ng/ml). Using the chemical analysis as the gold standard for identifying illegally treated animals, the histological examination of the thymus had a sensitivity of 100 per cent and a specificity of 85 per cent. The samples that were positive by chemical analysis had cortisol concentrations of less than 2.0 ng/ml, whereas the mean cortisol concentration of the negative samples was 10.3 ng/ml.

Administration of dexamethasone per os in finishing bulls. II. Effects on blood parameters used as indicators of animal welfare.

A set of hormonal, haematological and biochemical parameters was used to evaluate the physiological response and welfare status of 14 finishing Marchigiana bulls treated for 49 days with a low daily dosage (0.75 mg/head per day) of dexamethasone per os. Compared to the Control group, dexamethasone decreased cortisol concentrations (42.3 v. 5.7 nmol/l; s.e.d. = 4.17; P < 0.001), and led to the reversal of the leukocyte formula in the animals treated (P < 0.05). Total serum proteins (70.2 v. 73.9 g/l; s.e.d. = 1.55; P < 0.05), in particular ß1 globulins (7.5 v. 9.1 g/l; s.e.d. = 0.24; P < 0.01) and fibrinogen (199 v. 258 mg/dl; s.e.d. = 32.70; P < 0.05), increased as a consequence of treatment. Prolonged dexamethasone administration led the bulls to an apparently chronic stress condition. Moreover, the study indicated various blood parameters that might be used by health officials as effective tools in identifying beef cattle suspected of being illegally treated with dexamethasone.

Subacute rumen acidosis in lactating cows: an investigation in intensive Italian dairy herds.

Subacute rumen acidosis (SARA) represents one of the most important metabolic disorders in intensive dairy farms that affects rumen fermentations, animal welfare, productivity and farm profitability. The aim of the present study was to study the occurrence of SARA in intensive Italian dairy herds and to determine the relationship between diet composition, ruminal pH and short chain fatty acids (SCFA) concentration. Ten commercial dairy herds were investigated; twelve cows in each herd were selected randomly among animal without clinical signs of disease, with good body condition and between 5 and 60 day-in-milk (DIM), to perform rumenocentesis and obtain rumen fluid. Ruminal pH was determined immediately after sampling and concentration of SCFA in ruminal fluid was determined on samples after storage. An other objective of this research was to study in detail the effects of rumenocentesis on animal health: this study could confirm the extreme validity of this technique as ruminal sampling. Results were subject to anova and correlation analysis using SIGMA STAT 2.03. The results indicated the presence of SARA in three herds (more than 33% cows with rumen pH < 5.5), a critical situation (more than 33% cows with rumen pH < 5.8) in five farms and a normal rumen pH condition in two herds. In particular, dairy herds show on average SCFA concentration of 150, 145, 123 mmol/l for low pH, critical pH and normal pH herds respectively. There were not significant differences among diet composition even if herds with SARA showed a light discordance between initially chemistry composition and residual feed. In the affected herds it was not possible to understand the exact causes of SARA. Animal management seems to be one of the most important factors in developing SARA including total mixed ration preparation.

Near-infrared reflectance spectroscopy as a method to predict chemical composition of breast meat and discriminate between different n-3 feeding sources.

The objective of this study was to evaluate near-infrared reflectance spectroscopy (NIRS) as a tool to predict the physicochemical composition of breast meat samples of laying hens fed 4 different diets, a control and 3 diets enriched with different sources of n-3 polyunsaturated fatty acids: marine origin, extruded linseed, and ground linseed. Furthermore, NIRS was used as a tool to classify meat samples according to feeding regimen. Samples were analyzed chemically for DM, ash, protein, lipids, and fatty acid profile. Absorption spectra were collected in diffuse reflectance mode between 1,100 and 2,498 nm every 2 nm. The calibration results for the 72 meat samples were accurate in predicting DM, protein, lipids, and major fatty acids. Poor results were obtained for the calibration equations for ash, pH, color, and lipid oxidation parameters. Partial least squares discriminant analysis was developed to differentiate the breast meat samples that originated from hens fed the different diets. The performance of the discriminant models showed 100% correct classification between the control and the enriched diets. It was concluded that NIRS could be used for quality control predicting chemical composition of poultry meat and possibly some dietary treatments applied to the chickens.

The provision of drinking water to veal calves for welfare purposes.

Growth performance, behavior, physiology, forestomach development, abomasal lesions, and meat quality of veal calves fed a milk-replacer diet (No Water) were compared to those obtained from calves fed the same diet and provided with increasing amounts of drinking water (Water). Two groups of 69 Polish Friesian calves, balanced according to initial BW, were assigned to two water treatments in a 3 x 2 x 2 factorial arrangement that provided solid feed in addition to the milk-replacer diet (No solid feed, 250 g x calf(-1) x d(-1) of wheat straw or the same amount of beet pulp), and the adoption of two housing systems (individual stall vs group pen). The fattening trial lasted 160 d, and calves received drinking water starting from the 2nd wk of the study. The amount of drinking water was progressively increased from 3 to 8 L x cal(-1) x d(-1). Although not dehydrated, as shown by hematocrit and Na, K, and total protein hemoconcentration, calves consumed almost all the offered amount of water throughout the fattening period. Therefore, the water provided by the milk replacer alone, which ranged between 6 to 16 L calf d(-1), was not sufficient to satisfy the need of the animal. Drinking water did not affect the calves' growth performance but it reduced nonnutritive oral behavior throughout the fattening period. Based on these results, drinking water did not cover a shortage in the calves' water requirement but it played a role in environmental enrichment. Health status was similar between treatments, although water provision reduced the episodes of feed refusal. The measurement of chronic stress by ACTH challenge showed that the administration of drinking water would be advisable when calves are fed with small amounts of solid feed for well-being. Feces consistency and animal cleanliness were not affected by drinking water. At slaughter, forestomach development was similar between treatments, and drinking water did not affect the number of calves showing rumen hair-balls and abomasal lesions. No differences in color and other meat quality traits were observed between Water and No Water calves. Despite the lack of direct effects on productive traits, when water was available, the calves drank it, and positive effects were noticed on their nonnutritive oral behaviors and chronic stress indicators.

Glucose or essential amino acid infusions in late pregnant and early lactating Simmenthal cows failed to induce a leptin response.

To assess the leptin response to metabolic challenges, three Italian Simmental cows were infused for 6 h: with (a) saline (control); (b) glucose; and (c) amino acid solutions according to a 3 x 3 Latin square experiment. The infusions were carried out at the 36th week of pregnancy, and the second and 12th week of the following lactation. At each of the three infusion periods, blood samples were collected from the jugular vein before and 15, 30, 120, 180, 240, 300 and 360 min after the beginning of each infusion. All samples were analysed for leptin, insulin, glucagon, growth hormone (GH), glucose, non-esterified fatty acids (NEFA) and urea. The physiological phase of the cows significantly affected the basal concentrations of insulin, glucagon, urea and NEFA. The infusion of both glucose and the amino acid solutions did not affect leptin concentrations. Insulin response was significantly increased when animals were infused with the glucose solution and, within treatment, the greatest response was observed at the 12th week of lactation. The greatest glucagon response was observed when infusing the amino acid solution. Urea response to all treatments increased from the dry period to the 12th week of lactation. The GH and NEFA responses were not affected by treatments. The Multi Species radio-immunoassay used in this study showed a lower sensitivity for ruminant leptin which may partially explain the lack of significant leptin variations. However, it can be hypothesized that leptin variations around parturition can be affected by the negative energy balance, and leptin release is not acutely affected by glucose and amino acid availability. In addition, no short-term relationship were found between insulin, glucagon and GH and leptin release in Italian Simmental cows during the dry period and early lactation.

The provision of solid feeds to veal calves: II. Behavior, physiology, and abomasal damage.

The aim of this study was to evaluate the effects of the addition of two roughage sources (wheat straw and beet pulp) to the milk replacer diet of veal calves, in order to reduce stress and improve animal welfare. We allocated 138 Polish Friesian male calves to three different feeding plans: a milk replacer diet (Control), 250 g/d of wheat straw in addition to the milk replacer, or 250 g/d of dried beet pulp in addition to the milk replacer. Within each feeding treatment, 16 calves were individually housed and 30 were kept in group pens (five calves/pen). Several behavioral, physiological, and health welfare indicators were monitored throughout the fattening period, which lasted for 160 d. Abnormal oral behavior around the meals was higher in Control calves (P < 0.01), while its lowest level was observed in straw-fed calves. At the beginning of the trial, chewing was higher in calves receiving solid feeds (P < 0.001), but the difference from the Control gradually decreased and disappeared at wk 13 for calves fed beet pulp and at wk 17 for those fed wheat straw. At the end of the fattening period, no differences among treatments were found in the frequency of chewing. Regardless of the diet, self-grooming decreased with age and no relationship was observed between this behavior and the presence of rumen hairballs. Cross-sucking was performed with low frequencies (from 4.70% at wk 2 to 1.05% at wk 23 around the meals, and even lower far from the meals) and was not affected by the provision of roughage. The time in contact with the bucket during the whole day was higher in Controls, whereas calves fed wheat straw maintained a lower level of this activity until the end of the trial (P < 0.01). The calves fed wheat straw spent more time in contact with the feed trough (P < 0.001) than those fed beet pulp and Control calves. No differences were found in cortisol curves due to the feeding treatment. In calves fed beet pulp, most hematological measures statistically differed from the other treatments, possibly in response to the higher iron intake and(or) to the higher hemoconcentration, probably due to the administration of beet pulp as dried feed. The incidence of abomasal ulcers and erosions was increased by the provision of the solid feeds, particularly by a structured fiber source such as straw. A roughage source able to satisfy calves' behavioral needs and to improve digestive processes without damaging the digestive apparatus still has to be identified.

The provision of solid feeds to veal calves: I. Growth performance, forestomach development, and carcass and meat quality.

Growth performance, forestomach development, and carcass and meat quality of veal calves fed a milk replacer diet (Control) were compared to those obtained from calves fed the same liquid diet plus 250 g x calf(-1) x d(-1) of dried beet pulp or wheat straw. Three groups of 46 Polish Friesian calves, balanced according to initial BW, were assigned to the three dietary treatments in a fattening trial, which lasted 160 d. The provision of either solid feed did not affect the milk replacer intake. However, calves' ADG was increased (P < 0.01) only by feeding the beet pulp diet. The administration of both solid feeds improved calves' health status; calves fed solid feeds required fewer iron treatments for low hemoglobin and needed less medical treatments for respiratory or gastrointestinal diseases. In comparison to the Control calves, the provision of wheat straw and beet pulp increased iron intake throughout the fattening period by 41 and 130%, respectively. However, only calves fed beet pulp showed higher levels of hemoglobin and plasma iron concentrations (P < 0.05), whereas the same blood parameters were similar between Control calves and those fed wheat straw. At slaughter, both solid feeds led to empty forestomach weights heavier than those of Controls without reducing dressing percentage. The reticulorumen was heaviest in calves fed beet pulp, whereas wheat straw promoted omasal development. The administration of beet pulp resulted in a better carcass conformation than did the Control diet or wheat straw, but it had a detrimental effect on carcass color, which was graded as the darkest (P < 0.001). Consistent with this result, meat color of calves fed beet pulp was darker than that of Control calves and those fed wheat straw, because of the higher hematin concentration measured at the muscle level. No differences in carcass and meat color were observed between Control calves and calves fed wheat straw. The administration of solid feeds for welfare purposes does not always prevent the production of veal meat fulfilling the color standards required by the market. There is not a straight-forward relationship between a solid feed's iron content and the "redness" of veal meat, which should be related to the capability of the calves to use the iron provided by the roughage.

A blend of animal and cereal protein or fish meal as partial replacement for soybean meal in the diets of lactating Holstein cows.

Six replications in Experiment 1 and four replications in Experiment 2 of a 3 x 3 Latin square arrangement of treatments were used to compare soybean meal or soybean meal partially replaced with fish meal or a protein blend for response in intake, milk yield and composition, ruminal NH3 N, blood urea, and ruminal fermentation in lactating Holstein cows. The blend contained 30% corn gluten meal, 30% poultry by-products, 30% blood meal, and 10% feather meal. Periods were 28 d, and the first 7 d were used for adjustment. In addition to these protein sources, diets contained corn silage, alfalfa haylage, dried cracked corn, ground barley plus added fat, and a mineral and vitamin mixture. In Experiment 1, mean DMI was 24.4 kg, mean milk yield was 36.7 kg, mean fat percentage was 3.48%, and mean milk protein percentage was 3.06%; there were no significant differences. In Experiment 2, DMI was different for soybeans (22.6 kg) versus other sources (21.4 kg), but milk yield (32.1 kg) and fat (3.39%) and protein (2.87%) percentages did not differ among diets. In Experiment 1, ruminal NH3 N was greatest for cows consuming soybean diets (11.0 mg/dl) and lowest for cows consuming diets containing the protein blend (8.7 mg/dl). No differences in VFA were found. The lack of response to RUP can be explained by a rather high intake of a fermentable diet, which supplied sufficient absorbable AA according to the Cornell AA model.

Effect of slow-release somatotropin on the pattern of milk yield between and within injection intervals.

Thirty-six Alpine goats (10 primiparous and 26 multiparous) were used to study the milk yield response induced by recombinant bST administered in a sustained-delivery vehicle. Eighteen goats were injected with recombinant bST at 4-wk intervals from about 7 wk postpartum for 12 wk. Patterns of milk yield were highly affected by injection of recombinant bST. Milk yield of treated goats increased from day of injection, peaked between d 3 to 5 after treatment, and then decreased progressively. This pattern of response was well fitted by a curve containing two exponential terms: Yi = A x [exp(-k1di - exp(-k2di)], where Yi = daily relative response of treated goats to recombinant bST, computed as a deviation of least squares means of milk yield at day i (i = 0,..., 26) of the treatment interval from least squares means of milk yield at d 27 of the treatment interval; A = constant; k1 and k2 = response rate parameters; and di = day from treatment (i = 0,..., 27). Administration of recombinant bST also improved persistency of lactation, and the overall milk yield response to exogenous hormone was 300 g/d (13.9%) higher than the yield of untreated goats. This response was due to both the short-term response immediately after injection (53%) and the medium-term effect on lactation persistency.

The use of near infrared analysis for in situ studies.

This study evaluated the use of near infrared reflectance spectroscopy to estimate the chemical composition of feed residues after in situ ruminal incubation. Residues of three alfalfa hays (n = 93) and three alfalfa pellets (n = 93) obtained after ruminal exposure were analyzed for dry matter, crude protein, neutral detergent fiber, acid detergent fiber, and acid detergent lignin by wet chemistry and were also scanned with a near infrared monochromator instrument. A calibration was calculated that combined hay and pellet samples (n = 60). Validation tests were performed using the remaining feed residues. The coefficients of determination and standard errors (percentage of dry matter) of the validation tests for crude protein, neutral detergent fiber, acid detergent fiber, and lignin were of 0.95 and 0.92, 0.96 and 1.68, 0.95 and 1.56, and 0.99 and 0.48, respectively. Similar statistics were obtained using the SELECT algorithm of sample selection; a further 30% reduction was observed in the number of samples that were used for calibration. Kinetics of ruminal degradation and effective degradabilities that were calculated based on chemical composition of the residues as determined by wet chemistry or estimated by near infrared reflectance spectroscopy were not significantly different for 68 out of 71 means. Differences in ruminal kinetics caused by the different forages were also unaffected by method of residue analyses. Near infrared reflectance spectroscopy allowed a reduction in the number of necessary laboratory analyses of feed residues without affecting the results of in situ studies.

In situ ruminal disappearance of essential amino acids in protein feedstuffs.

Four protein sources were incubated in situ to estimate AA disappearance. Bags containing either soybean meal, corn gluten meal, herring meal, or meat meal were washed in water or suspended in the rumen of two Holstein cows for 8, 12, 16, 24, 48, 72, and 120 h. Cytosine, a bacterial marker for microbial contamination, was used to correct the essential AA profile for microbial contribution to determine the residual essential AA composition of the protein sources after incubation. Ruminal disappearance of individual essential AA was different among feedstuffs. Relative to original feed protein, soybean meal and corn gluten meal decreased the concentration of specific essential AA in the RUP. Concentration of all essential AA, except Arg and His, increased in undegraded meat meal protein. The difference between original and residual AA concentrations in herring meal approached statistical significance. Use of the original AA profile of the feed protein to predict essential AA available for absorption is not accurate because accuracy differs with sources.

Observations on in situ degradation of forage cell components in alfalfa and Italian ryegrass.

The rate and extent of degradation of forage feed fractions contained in alfalfa and Italian ryegrass hays were determined. Nylon bags filled with 4 g of each forage were suspended in the rumen of two cannulated cows immediately before feeding and incubated for 10 different times (0, 2, 4, 8, 12, 24, 48, 72, 120, and 168 h). The alfalfa hay, which had lower NDF, showed a lower extent, but a higher rate, of NDF degradation than the Italian ryegrass (41.1 vs. 59.8% and 4.64 vs. 2.91%/h, respectively). Alfalfa cell walls were degraded more rapidly than Italian ryegrass even though their lignin content was higher. The hemicellulose fermentation of alfalfa showed a longer lag time (13 h) and an undegradable fraction nearly twice that for Italian ryegrass (63.3 vs. 37.1%). Cellulose from alfalfa was degraded at a higher rate than NDF or ADF, indicating that cellulose may be the primary site of hydrolysis of the cell wall in the rumen. Calculations based on in situ degradability indicate that alfalfa can have a higher inclusion than Italian ryegrass in diets for dairy cows because of lower NDF and greater availability of cell contents.