Animal-based measures for welfare assessment

Animal welfare assessment can't be irrespective of measures taken on animals. Indeed, housing parametersrelatedtostructures, designandmicro-environment, evenifreliable parameters related to structures, design and micro-environment, even if reliable and easier to take, can only identify conditions which could be detrimental to animal welfare, but can't predict poor welfare in animals per se. Welfare assessment through animal-based measures is almost complex, given that animals' responses to stressful conditions largely depend on the nature, length and intensity of challenges and on physiological status, age, genetic susceptibility and previous experience of animals. Welfare assessment requires a multi-disciplinary approach and the monitoring of productive, ethological, endocrine, immunological and pathological param- eters to be exhaustive and reliable. So many measures are needed, because stresses can act only on some of the mentioned parameters or on all of them but at different times and degree. Under this point of view, the main aim of research is to find feasible and most responsive indicators of poor animal welfare. In last decades, studies focused on the following parameters for animal wel- fare assessment indexes of biological efficiency, responses to behavioral tests, cortisol secretion, neutrophil to lymphocyte ratio, lymphocyte proliferation, production of antigen specific IgG and cytokine release, somatic cell count and acute phase proteins. Recently, a lot of studies have been addressed to reduce handling and constraint of animals for taking measures to be used in welfare assessment, since such procedures can induce stress in animals and undermined the reliability of measures taken for welfare assessment. Range of animal-based measures for welfare assessment is much wider under experimental condition than at on-farm level. In welfare monitoring on-farm the main aim is to find feasible measures of proved validity and reliability to be taken on a large number of animals in a reasonable time. Human-animal interaction, body condition score, skin and hair conditions, lameness and injuries, cleanliness, and abnormal behaviors are regarded as the most reliable animal-based indicators for welfare assessment on-farm

Technical Note: A Simple Salting-Out Method for DNA Extraction from Milk Somatic Cells: Investigation into the Goat CSN1S1 Gene

Abstract In this study, a sensitive, rapid, and toxic solvent-free method to extract DNA from milk somatic cells was implemented for characterization of the goat α S1 -casein gene ( CSN1S1 ). Methods reported for purification of DNA from milk often involve organic extraction, overnight incubation, or use of expensive commercial kits. The present method was implemented for goat milk and is based on a salting-out protocol. The method yielded an amount of DNA suitable for PCR-RFLP without the need for sample enrichment. The PCR-RFLP of DNA extracted from milk produced amplified and digested products of correct size, comparable with those obtained using PCR-RFLP of DNA extracted from blood. Therefore, milk can be used as an alternative source of DNA, making sample collection easier and reducing stressful practices such as capture, handling, and venipuncture in animal management

Changes occurring in immune responsiveness of single- and twin-bearing Comisana ewes during the transition period.

Changes induced by twin and single lambing in the immune response of 16 periparturient Comisana ewes were studied. Cell-mediated immune responses were evaluated by means of skin tests performed from 3 wk before and up to d 35 after parturition. At d 21 and 7 before lambing, the sheep received an intramuscular injection of the antigen keyhole limpet hemocyanin (KLH), to which the animals had not been previously exposed, to determine their humoral immune response. Starting 3 wk before lambing and up to d 35 postlambing, the ewes were sampled to determine the plasma concentrations of anti-KLH antibody (IgG), IL-6, and IL-1 beta. From parturition through d 35 postpartum, individual milk samples were collected for determination of anti-KLH IgG titers and IL-6 and IL-1beta concentrations by means of a capture ELISA. The number of lambs born affected IL-6 concentrations in ewe plasma; IL-6 secretion always was higher in ewes birthing twins than in single-lambing ewes. Apart from the number of lambs born, the concentrations of plasma IL-6 in ewes were higher at lambing than at d 21 antepartum and at d 35 postpartum. An interaction of number of lambs born x time of sampling was observed for plasma antibody titers to KLH. The IgG concentrations were significantly higher in single-bearing ewes than in twin-bearing ewes before parturition and were very similar across groups after parturition. A time effect was found for the cell-mediated immune response and for anti-KLH IgG concentrations in milk, such that at parturition, cellular responses were lowest, and the anti-KLH IgG concentration was highest. A significant correlation was found for IgG titers to KLH in plasma and milk. Results indicate that IL-6 concentrations in blood can be considered a reliable indicator of stress connected to lambing and that the mammary gland is a microenvironment unrelated to blood stream with respect to interleukins expression. In contrast, a relationship was found for the IgG secretions in milk and blood, which suggests that the assessment of humoral immune status may be combined with milking routine in dairy animals

Technical note: immunomagnetic procedure for positive selection of macrophages in ovine milk.

A simple immunomagnetic procedure was developed to select macrophages from ovine milk by using a non-specific magnetic positive separation technique. Samples of ewe bulk milk were collected during early, mid, and late lactation; milk samples were centrifuged at 2,000 x g for 30 min at 4 degrees C; the fatty fraction and supernatant were removed, and each pellet was dissolved in 500 microL of pH 7.4 phosphate buffered saline + 0.02% NaN(3). Cells were targeted for selection by using mouse-IgG anti-ovine macrophages. Several trials, testing 2 different fluorochrome-conjugated antibodies [i.e., mouse anti-human CD14:R-Phycoerythrin (RPE) (MCA1568PE, Serotec) and F(ab')2 rabbit anti-mouse IgG:RPE (STAR12A, Serotec)] and 3 different labeling procedures, were performed to evaluate the purity of samples by flow cytometry. A morphological test was carried out by direct microscopic count in enriched fraction smears stained with May-Grünwald-Giemsa stain to confirm the presence of macrophages. The method described in the present technical note can be considered an innovative application to obtain a single-cell population of high purity selected from all the somatic cells in milk

Airspace effects on the yield and quality of ewe milk.

Abstract Three groups of 12 midlactating Comisana ewes were housed in separate rooms of the same building and assigned to treatments of low (LV, 4.1m 3 ), medium (MV, 5.6m 3 ), or high (HV, 7.3m 3 ) airspace/animal. The concentrations of airborne microorganisms in the experimental rooms were measured twice weekly at 0930 and 1630. Ewe milk yield was recorded daily. Individual milk samples were analyzed weekly for milk composition, coagulating properties, somatic cell concentration (SCC), and polymorphonuclear neutrophil leukocyte count (PMNLC), and fortnightly for bacteriological characteristics; samples with more than 10 6 somatic cells/ml were cultured for mastitis-related pathogens. The LV and MV treatments resulted in higher relative humidity and air concentrations of staphylococci than the HV treatment. Greater amounts of air mesophilic bacteria were also found in the LV than in the HV room. Ewes in the HV group gave greater yields of milk than those in the LV and MV groups. LV milk also had a lower casein content than HV milk. Significant interactions of treatment x time were found for milk protein and fat content as well as for clotting time and clot firmness, with LV milk having the poorest composition and deteriorated renneting ability during the last 3 wk of the trial. The HV ewes had lower SCC and PMNLC and psychrotroph counts in their milk than LV and MV ewes and smaller amounts of mesophilic bacteria and fecal coliforms than LV animals. Subclinical mastitis occurred in two ewes of the LV and one of the MV groups, while no cases were recorded in the HV group. Results suggest that airspace is a critical factor in dairy sheep housing and indicate that a volume allocation of less than 7m 3 /animal may adversely affect the performance and health of the lactating ewe

Indigenous enzymes and leukocyte in sheep milk are markers of health status and physiology of the mammary gland

Plasmin, plasminogen and plasminogen activator in ewe bulk milk were not significantly affected by stage of lactation probably as a consequence of the good health of the ewe udders throughout lactation as indicated by SCC which never exceeded 600,000 cells/mL. Elastase content increased significantly during lactation whereas cathepsin showed the highest content in mid lactation. Changes in macrophages and neutrophyls levels in ewe bulk milk during lactation were also investigated. Macrophages minimally contributed to leukocyte cell count in milk and had the highest levels at the beginning of lactation. An opposite trend was recorded for polymorphonuclear leukocytes (PMNL) that increased throughout lactation showing the highest value in late lactation. The increase of PMNL percentage and elastase content in milk, in spite of relatively low SCC, suggests that PMNL and elastase underwent a physiological increase associated to the remodelling of mammary gland in late lactation

Effects of graded noise levels on behavior, physiology and production performance of intensively managed lambs

The aim of the experiment was to assess the effects of graded noise levels on behavioral, immune and cortisol respons- es, and on production performance of forty precocious Merino lambs. The experiment lasted 42 days and was preceded by a two-week adjustment period. The animals were divided into 3 test groups and a control group of 10 each, which were housed in 12 m2 sound-proof rooms. The lambs in the test groups were exposed to recordings of high-speed motor vehicle traffic noise for 8 hours a day. The frequency range of the recordings was 100 to 6,300 Hz, while the loudness levels were different in the three experimental rooms: group A = 95 dB, group B = 85 dB, and group C = 75 dB. During the same 8 hours the control group was exposed to a background noise of 42-44 dB. For the rest of the day, the ambi- ent noise level in the animal building was 35-40 dB. Behavioral traits of lambs were recorded at the end of the adjust- ment period and then at 14d, 28d and 42d of the experiment. At the same intervals, the plasma levels of glucose, total cholesterol, triglycerides, total proteins, albumins and cortisol were determined. Cell-mediated immune response to phy- tohemagglutinin (PHA) was determined at the beginning and at days 21 and 42 of the trial. Live weights of lambs were measured at the beginning of the trial and then at two-week intervals during the trial. After slaughtering, the carcass weights and the dressing percentage were determined. The experimental treatment determined a significant increase in the walking time (P < 0.01) and a reduction of plasma cortisol levels (P < 0.01) in the samples taken on days 28 and 42 of the trial in groups A and B. The lambs in the test groups exhibited a slower growth rate (P < 0.01) than the ani- mals of the control group, whereas a deterioration of feed efficiency was observed only in group A (P < 0.01). There were no differences amongst the groups in terms of cell-mediated immunity and blood metabolites. Results suggest that expo- sure to noise has a detrimental effect on some biological functions in lambs but it does not severely impair animal well- being, at least when there are no concomitant conditions of physical and social discomfort

Quality of donkey bresaola

Nutritional and organoleptic properties were evaluated in bresaola of cow and don- key meat and, moreover, was made a comparison between these two products. The cured products were made from semitendinosus muscle taken from 8 female donkeys and 8 cows. Donkey bresaola showed lower SFA percentage (P<0.05) and higher PUFA content (P<0.01) with higher fatty acids ω 3 (P<0.01) than beef bresaola. Donkey bresaola showed higher content of protein (P<0.01) and essential amminoacids (P<0.05) than beef bresaola. No differences were found for sensorial properties. Our in- vestigation demonstrates the possibility of trasforming donkey meat into a product similar to that of beef with a high nutritional value

Consumer acceptance and sensory evaluation of Monti Dauni Meridionali Caciocavallo cheese

Twelve Caciocavallo cheeses were collected from 6 factories (A, B, C, D, E, F) located in the Monti Dauni Meridionali area (Southern Italy) that adopted different protocols for cheese production. A total of 160 consumers were involved in the sensory evaluation of Caciocavallo cheese after 180 d of ripening. Cheese attributes were used to describe the flavor, texture, and appearance of cheeses. The highest scores for the shiny attribute were assigned to cheeses B, C, and E, whereas color intensity was the highest in cheeses B, D, and F. Strength, salty, and piquant attributes were higher in cheeses F and A because of the use of raw milk (F), rennet paste (A), and percentage of salt in the brine (A, F). Consumers perceived a more granular structure during the second half of chewing of Caciocavallo cheese F, as evidenced by the highest value for the grainy attribute. A positive correlation was found between overall flavor and odor intensity and water-soluble nitrogen, low molecular weight peptides, and free fatty acids and between piquant and butyric and caproic acids. A principal components analysis applied to the sensory attributes accounted for 65% of the total variance. The score plot showed that cheeses F and A were located in a well-defined zone of the plot, with cheeses in this zone displaying higher levels of strength, piquant, and salty attributes. The preference test assigned 40% of the preference to Caciocavallo cheese A, 38% to cheese F, 9% to cheese E, 8% to cheese D, and 7% to cheeses B and C. Sensory evaluation of Monti Dauni Meridionali Caciocavallo cheeses is a useful analysis to highlight the principal attributes able to influence consumers' liking that are related to biochemical features of the cheese

Effects of Somatic Cell Count and Stage of Lactation on the Plasmin Activity and Cheese-Making Properties of Ewe Milk

The experiment was conducted from March to July 2002 using 5 intensively managed flocks of Southern Italy. In each flock, 2 groups of 50 ewes were created. The groups were designated LSCC (low somatic cell count [SCC]) when their milk SCC was lower than 500,000/mL and HSCC (high SCC) when their milk SCC was higher than 1,000,000/mL. Bulk milk and whey samples were analyzed for fat, total protein, lactose, casein, and whey protein contents. Renneting properties of milk were also determined. Moisture, NaCl, and nitrogen fractions were determined in fresh cheese curds. In addition, plasmin (PL) and plasminogen (PG) activities in milk and cheese were monitored. The proteolytic activity of plasmin by urea-polyacrylamide gel electrophoresis and the white blood cell (WBC) differentials were determined. The HSCC resulted in higher pH values in milk and in higher moisture and lower fat contents in fresh cheese curds. Moreover, a lower recovery of fat and whey proteins was obtained from the HSCC than from the LSCC raw milk. The crude protein and casein contents were higher in the HSCC than in the LSCC curds during early and midlactation; an opposite trend was observed in late lactation. Plasmin and PG activities underwent more marked fluctuations in the LSCC than in the HSCC curds through lactation. The results of this experiment demonstrate that the PL activity in ewe milk is markedly influenced by the SCC, although SCC is not the only parameter for predicting PL and PG evolution in ewe milk. The LSCC milk resulted in a higher proteolytic potential of Canestrato pugliese cheese curds