Short communication: Endoplasmic reticulum stress gene network expression in bovine mammary tissue during the lactation cycle.

Abstract The endoplasmic reticulum (ER) has a crucial role in cellular metabolism. Recent studies in nonruminants discovered that components of the ER stress pathway, induced during the unfolded protein response, play critical roles in regulating lipogenesis. The bovine mammary gland faces extreme metabolic stress at the onset of lactation due primarily to the increase in flux through pathways associated with milk fat and protein synthesis. Our objective was to study, via quantitative real-time PCR, the expression of the ER stress pathway components P58IPK , PERK , XBP1 , ATF4 , ATF3 , ATF6 , CHOP , MBTPS1 , GRP94 , and BiP in mammary tissue (n=7 cows × 5 time points) collected at −15, 1, 15, 60, and 240 d relative to parturition. Expression of P58IPK and ATF4 increased to a peak at d 60, followed by a decrease by d 240 postpartum. Despite the decrease in expression by 240 d, P58IPK remained higher than prepartal levels (d −15). Expression patterns of ATF3 and CHOP were similar and peaked at d 15, followed by a decrease through d 240, at which point CHOP expression was still greater than prepartal levels. The sharp increase in milk production postpartum (d 15) as well as apoptosis during late lactation (240 d) may have induced a pseudo unfolded protein response state. This is supported by the similar expression patterns of P58IPK and PERK . In the context of lactation, however, transcriptional changes in the ER stress pathway at different stages of the lactation cycle are a normal aspect of the tissue's adaptation to the changing physiological state

Plasma α-tocopherol content and its relationship with milk somatic cells count in Italian commercial herds.

This work was aimed to investigate relationship between plasma vitamin E concentration and milk somatic cell count in healthy cows in commercial herds. 49 multiparous cows from two commercial dairy herds were monitored from the day of dry off until 90 DIM. BCS was assessed and blood samples were collected at dry off, day 0, 30, 60 and 90 postpartum. Plasma was analyzed for α-tocopherol content. Quantification of NEFA, BOHB, Zn and Se was performed in serum samples. Milk production and composition was obtained from routinely test-day of Italian milk producers association. Somatic Cell Score (SCS) was calculated and included in the dataset. Analysis of data was performed using MIXED repeated and CORR procedures of SAS.We did not observe a correlation between plasmatic vitamin E and somatic cell score, and this can be explained by the low level of somatic cell score (averages 1.64 and 1.26). The lowest value of vitamin E was observed at parturition (1.64 µg/ml and 1.95 µg/ml). A significant (P<0.01) negative (-20%) correlation was observed between NEFA serum content and α-tocopherol plasma concentration. Serum selenium content was positively correlated (+42%, P<0.0001) to zinc concentration. Grouping cows on the basis of their plasma α-tocopherol content higher or lower than 3 μg/mL at dry off, SCS at 30 and 60 DIM tended to be higher in lactating animals with lower content of α-tocopherol (1.12 vs. 1.72, P=0.18 at 30d; 0.92 vs. 1.72, P=0.07 at 60d). However, plasma α-tocopherol content at dry off could be usefully correlated with somatic cell count in fresh cows

Effect of live yeast (Saccharomyces cerevisiae) administration on apparent digestibility of horses

Six mares were divided into two groups of three subjects each in a crossover design on the basis of body weight and age: Control(C)fedabasaldiet,Treatment(T)fedabasaldietand2 Control (C) fed a basal diet, Treatment (T) fed a basal diet and 2 2 g/head/d of live yeast ((S. cerevisiae 4.6x1010 CFU/day).Theexperimentaldesignwasdividedintotwo ).Theexperimentaldesignwasdividedintotwo . The experimental design was divided into two periods named period 1 and period 2 respectively of 35d each, and consisting of 3 different phases. Dur- ing each period all animals were subject to an adaptation phase of 14d (phase1); during phase 2 (18d) and phase 3 live yeast was administered (T) or not (C). Phase 3 consisted in a three days individual fecal collection period all the groups, in order to determine dry matter, organic matter, crude protein, crude fat, NDF and ADF apparent digestion rates using acid insoluble ash (AIA) as internal marker (Bergero et al., 2005). Results obtained evidenced as the administration of S. cerevisiae to mature horses resulted in increased digestibility of dry matter, organic matter, NDF, and ADF

Feed efficiency of Italian Holstein dairy heifers in the genomic era – preliminary results

Genetic selection of dairy cattle has classically been based on fertility and productivity (kilograms of milk, protein, fat). In recent years a growing interest in characters related to health and efficiency of the animals has taken hold. The selection of animals with a high feed efficiency can bring benefits in terms of health, productivity and environmental impact. A first batch of 16 Holstein heifers (mean age 12.63 ± 2.90 months) was selected and housed in a tie-stall of the Centro Zootecnico Didattico Sperimentale (CZDS) of Lodi for the whole length of the trial (35 days). Blood samples were collected and sent to the lab to perform genotyping of the animals. Heifers were fed a composed ration of sorghum silage and total mixed ration, delivered daily on individual feeders to ensure each animal continuous access to the feed. Animals had free access to the water. Feed intake was obtained weighing the supplied ration and the residual the day after. Individual body weight (BW), body condition score (BCS), hearth girth (HG) and height at the wither (W) were measured weekly. Feed Conversion Ratio (FCR) and Residual Feed Intake (RFI) were calculated using Dry Matter Intake (DMI) data. Fecal samples were collected weekly to assess any relation between feed efficiency and acidic profile of the feces. Preliminary results show an average daily gain of  941 ± 206 grams, an increase of mean BW from 374.63 ± 17.06 (day 0) to 407.56 ± 17.06 kilograms (day 34), an increase of mean BCS from 3.97 ± 0.11 (day 0) to 4.19 ± 0.11 (day 34), an increase of mean HG from 169.25 ± 2.83 (day 0) to 177.41 ± 2.83 centimeters (day 34), an increase of mean W from 127.28 ± 1.23 (day 0) to 132.69 ± 1.23 centimeters (day 34)

Can lipid supplementation modulate inflammatory state and immune response in periparturient goats? A case study on hepatic and adipose miRNA expression

Saturated or unsaturated fatty acids compounds were administered to second-parity twins-diagnosed alpine dairy goats. Experimental groups were fed either calcium stearate (ST, n.7), fish oil (FO, n.8) or a control diet without any fat supplement (C, n.8) from one wk before (30g/head/d of fatty acids) to three wks after kidding (50g/head/d of fatty acids). ST provided 26% C16:0 and 69.4% C18:0 while FO provided 10.4% EPA and 7.8% DHA. Both ST and FO diets were formulated to be isocaloric and isonitrogenous, with the same calcium content. Previous obtained results on metabolic, productive parameters and mRNA expression of genes related to lipid metabolism and inflammatory response let us to consolidate the hypothesis that in goat lipogenesis is reduced across the transition period, if compared to cow, moreover FO can postpone or reduce lipomobilization. Based on these previous results, miRNA expression was performed on the same hepatic and adipose biopsies (collected on day -7 and 7 and 21 from kidding), as a new perspective in controlling cellular pathways, implicated in adipogenesis and metabolic and endocrine functions. We examined miR-26b and 155 for the infiltration of immune cells, miR-99a, 145 and 221 for the inflammation and lipolysis, miR-143 and 378 for pro-adipogenic function. MIXED and GLM procedures of SAS software were used for statistical analysis. No diet effect, but a time effect for miR-155 and a tendency for miR-221 were found. The increase of their expression over the time after kidding let us to speculate that goats have to face a postponed and more contained inflammation due to the lipolysis, when compared to cows

Nuclear factor E2-related factor 2’s activation in transgenic mice fed with dosage of saturated or unsaturated fatty acids using in vivo bioluminescent imaging

To counteract oxidative stress cells developed several mechanisms, including the transcription factor Nuclear Factor E2-related factor 2 (Nrf2). The aim of the study was to evaluate the activation of Nrf2 in transgenic mice fed saturated or polyunsaturated fatty acids and the anti-inflammatory effect of estrogens on organism. Forty-eight ARE CRE OMO reporter mice were divided into 3 groups, consisting of 16 animals, based on presence/absence of estrogens (ovariectomized or sham female, OVX - SH; male, MA). Each group was further split in 4 subgroups of 4 animals each and fed different diets (7.5% lard, 7.5% tuna oil, 20.0 % lard and 20.0% tuna oil). Two times a week animals were anaesthetized and injected i.p. with 100µL luciferin 15 min before the imaging session. Using the Living Image Software, photon emission was mapped for selected body areas. On day 70, animals were sacrificed after a challenge with Sodium Arsenite. Specific organs were dissected and immediately subjected to ex vivo imaging session. MIXED and GLM procedures of SAS software were used for statistical analysis. Dietary treatments did not affect body weight and feed intake as well as Nrf2 expression in both pre- and post-challenge phases, with the exception of the abdominal region (P=0.031 pre-challenge); in this area, during the pre-challenge phase, OVX showed lower Nrf2 activation (P<0.001). Ex vivo results outlined a significant effect of the challenge on all the considered organs (P<0.001), while OVX subjects had higher Nrf2 expression on urinary bladder and kidney (P<0.05) and high fat diet increased Nrf2 in urinary bladder (P<0.05). The present trial shows how saturated or polyunsaturated fatty acids supplementation in the diet do not exert significant effects on oxidative stress in mice, but confirms the protective role of estrogens under physiological condition

Microbiological characteristics of poultry meats - Results of inspections carried out in the province of Milano, Italy

Examinations were conducted in terms of microbiological quality/quantity (TMC, Coliforms, E. coli, S. aureus, Sulphate-reducing Clostridia, B. cereus, Salmonella spp. and Lysteria spp. and Campylobacter spp.) on 240 poultry meat samples (chicken, turkey and quail) acquired pursuant to the standards set by the Regional Plan of programming and coordination in the field of operations concerning official inspections of Lombardia animal origin and by a few private companies for self-inspection. The TMC was consistently low and in line with reports in the literature, as was the case with coliforms, E. coli, S. aureus, sulphatereducing Clostrides and B. cereus. In the case of Salmonella spp., only 5 samples tested positive: one for S. typhimurium and one for S. enteritidis (chicken); only one sample from turkey tested positive for S. blokley, and two out of five samples analysed from quail tested positive result for S. typhimurium. About 3% of the samples analyzed tested positive for Listeria monocitogenes, but they were within the legal limits. Research on Campylobacter thermophiles has involved only 50 samples, of which only 5 have tested positive. These results confirm the high quality of hygiene and cleanliness of poultry meat, in accordance with that reported in the national literature and with respect to EU norms

Transcriptional regulation of lipid metabolism and inflammation in transition dairy goats by fish oil and stearate

To better understand the interaction between saturated or unsaturated fatty acids and its effect onexpression of genes involved in subcutaneous adipose tissue metabolism, 23 second parity alpine dairygoats were enrolled in the experiment and fed either a non fat-supplemented basal diet (C; n=8), thebasal diet supplemented with stearic acid (ST; n=7) or the basal diet supplemented with fish oil (FO;n=8). 30g/head/d supplemental fatty acids during the dry period and 50g/head/d during lactation weredelivered starting one week before parturition up to 21 days in milk. Subcutaneous adipose tissuesamples were harvested at day -7, 7 and 21 relative to kidding and mRNA levels of genes involved ininflammation were measured via qPCR. Data were analyzed using the MIXED procedure of SAS. Nosignificant effects for treatment were observed, however eight genes were significant for time. HP andSAA3 expression peaked at day 7 postpartum, to then return at prepartum level around 21 d relative tokidding, while IL8, IL10, and IL18 expression constantly increased along the transition period. Vice versa,expression of IL1β, IL6R, and RXRA decreased in response to kidding, with a subsequent increase at day21. The obtained results led us to hypothesize that goats face a postponed lipomobilization afterkidding, probably related to their reduced production. The next step will involve the analysis of miRNArelated to immune cell infiltration, adipocyte inflammation and lipolysis and positive regulation ofadipogenesis to better understand the complex network of lipid metabolism in periparturient goats

The beneficial role of Probiotics in monogastric animal nutrition and health

The practice of probiotics in farm animals' health and production has increased significantly over the last 15 years. Probiotics are defined as live microorganisms that can confer a health benefit for the host when administered in appropriate and regular quantities. The isolation and identification of microorganisms is the first step in the selection of potential probiotics from gut, feces and milk of respective animals. The present molecular techniques mainly genomic and proteomic-knowledge based are employed to identify, characterize probiotics. The ability to examine fully sequenced genomes has accelerated the application of genetic approaches to elucidate the functional roles in the selection of new and specific probiotics. Identification of suitable probiotics may prove to be the next step to decrease the risk of intestinal diseases and reduce specific microbial disorders, as well as demonstrating their role in the production performance of animals, safety and wholesomeness of animals' meat evidencing consumer's protection. The mechanisms of action of probiotics include the inhibition of pathogen growth by competition for nutritional sources and adhesion sites, secretion of antimicrobial substances and toxin inactivation. Consequently, the primary interest in the application of probiotics has been in the prevention and treatment of gastrointestinal infections and antibiotic-associated animals' diarrheal diseases. In this review, the most important benefits of probiotics upon the gastrointestinal microbial ecosystem in monogastric animals (equines, pigs, veal calf and poultry) are described, as well as their implications in terms of animal nutrition and health. Additional knowledge on the possible mechanisms of action is also provided

Cap rock efficiency of geothermal systems in fold-and-thrust belts:evidence from paleo-thermal and structural analyses in Rosario de La Frontera geothermal area (NW Argentina)

Cap rock characterization of geothermal systems is often neglected despite fracturing may reduce its efficiency and favours fluid migration. We investigated the siliciclastic cap rock of Rosario de La Frontera geothermal system (NW Argentina) in order to assess its quality as a function of fracture patterns and related thermal alteration. Paleothermal investigations (XRD on fine-grained fraction of sediments, organic matter optical analysis and fluid inclusions on veins) and 1D thermal modelling allowed us to distinguish the thermal fingerprint associated to sedimentary burial from that related to fluid migration. The geothermal system is hosted in a Neogene N-S anticline dissected by high angle NNW- and ENE-striking faults. Its cap rock can be grouped into two quality categories: ● rocks acting as good insulators deformed by NNW–SSE and E–W shear fractures, NNE-SSW gypsum- and N-S striking calcite-filled veins formed during the initial stage of anticline growth. Maximum paleo-temperatures (<60°C) were experienced during deposition to folding phases. ● rocks acting as bad insulators deformed by NNW-SSE fault planes and NNW- and WNW-striking sets of fractures associated to late transpressive kinematics. Maximum paleo-temperatures higher than about 115°C are linked to fluid migration from the reservoir to surface (with a reservoir top at maximum depths of 2.5 km) along fault damage zones. This multi-method approach turn out to be particularly useful to trace the main pathways of hot fluids and can be applied in blind geothermal systems where either subsurface data are scarce or surface thermal anomalies are lacking