Effects of exogenous glucoamylase enzymes alone or in combination with a neutral protease on apparent total tract digestibility and feces D-Lactate in crossbred angus bulls fed a ration rich in rolled corn

The aim of this study was to evaluate the effect of two glucoamylases (GA) and the combination of one GA with a neutral protease on apparent total tract digestibility in beef bulls fed a total mixed ration (TMR) rich in rolled corn. Sixteen Angus beef bulls (266 ± 4.9 kg of initial BW, and 182 ± 1.7 d of age) were distributed in 4 blocks, each block consisted of 4 animals balanced by BW. The experimental design was a 4 × 4 Latin square (4 blocks and 4 periods, 2 w per period). Four treatments were tested; (1) control, (2) GA preparation from Trichoderma reesei (TrGA); (3) GA from Aspergillus fumigatus (AfuGA); (4) AfuGA in combination with a neutral protease from Bacillus amyloliquefaciens (BamPro). Apparent total tract digestibility and fecal D-lactate concentration were analyzed. Enzyme supplementation, regardless of enzyme type, increased apparent total tract digestibility of dry matter (from 66.7% to 73.1% ± 2.01), and starch (from 74.7% to 81.8% ± 2.25), without affecting feces D-lactate concentration. Irrespective of glucoamylase type, glucoamylase supplementation improved apparent digestibility of dry matter and starch, and the addition of a protease did not have additional benefits on nutrient digestibilityinfo:eu-repo/semantics/publishedVersio

Vergence and accommodation cues in stereo-localization during the Small-In Large-Out (SILO) effect

A typical procedure in vision therapy is the use of Quoits vectograms to train fusional vergence ranges by improving stereo-localization, which is the ability to correctly locate the target stimulus in space. With this procedure, the Small-In Large-Out (SILO) effect is usually reported in patients with normal binocular vision and accommodation. In this study, the influence of vergence and accommodation cues, as determined with the accommodative-convergence over accommodation (AC/A) ratio, to correctly locate the Quoits vectograms in space was investigated. Twenty participants, aged 29.2 ± 2.8 (mean ± standard deviation) years, without amblyopia or strabismus, were recruited. A geometrical formula was obtained to calculate the theoretical distance to the target stimulus for different vergence demands. Theoretical values were compared with measured distances to the perceived stimuli and stereo-localization accuracy was determined. Stereo-localization accuracy was significantly worse at 10¿ Base In vergence demand (p < 0.001). A statistically significant positive correlation was found between AC/A ratio and stereo-localization accuracy (i.e., worse accuracy) at 10¿ Base Out vergence demand (rho = 0.446, p = 0.049). These findings highlight that AC/A ratio may be a secondary cue for stereo-localization when using vectograms in which the SILO effect is manifest. These results assist in the understanding of the physiological basis of vision therapy procedures.Peer ReviewedPostprint (author's final draft

Short communication: A milk replacer aversion model in calves to test flavour-masking effects

This study aimed to evaluate the availability of two sensory additives (SAs) to masque non-palatable ingredients commonly present in milk replacers (MRs). Both SAs were based on a synergistic mixture of non-nutritive flavouring substances with four taste categories (sweet, umami, sour, and salty); the only difference between them was the inclusion of polyols, which were not included in SA2. Two experiments were conducted for this purpose. In Experiment 1, an MR aversion model was developed using 24 Holstein male calves (7 ± 0.9 d of age and 43.3 ± 1.39 kg of body weight [BW]). In the first two weeks of the study, calves were fed increasing amounts of MRs until the animals were able to consume 8 L/d at 15% dry matter (DM) concentration for two feedings per day. Thereafter, MR aversion was induced in half of the calves by adding a mixture of bitter taste feed additives (Bittermix) from days 14 to 22 of the study (aversion week). The daily MR intake and eating rate were recorded from the two previous days of the challenge and during the aversion week. In Experiment 2, the same model was used with 37 Holstein male calves (6 ± 0.9 d of age and 40.2 ± 1.40 kg of BW). Owing to health issues in this experiment, the aversion week was postponed at 38.5 ± 1.12 d of age until the animals were able to consume at least 90% of 8 L/d at 12.5% DM concentration. The aversion test was performed as follows: no supplementation (CTRL), Bittermix at 30 g/kg of dry MR (BM), BM plus SA1 at 2 g/kg of dry MR (SA1), and BM plus SA2 at 2 g/kg of dry MR (SA2). Data were analysed with a generalised mixed model that accounts for the fixed effects of MR supplementation, the meals during the aversion week, the interaction of the MR supplementation and meals, and a calf as a random effect. In the aversion test of Experiment 1, calves that were fed Bittermix showed an increase in the incidence and amount of MR refusals (P < 0.05) compared with CTRL calves. In Experiment 2, the incidence of refusal and time devoted to consuming MR were similar in all treatments. However, the eating rates were greater (P < 0.01) in CTRL and SA2 than in BM and SA1. Adding a mixture of bitter tasting feed additives in the MR changed the feed intake parameters of the calves. The addition of a specific sensory additive, namely, SA2, reversed the negative effects caused by the bitter products.info:eu-repo/semantics/publishedVersio

Assessment of the Impact of the Recombinant Porcine Reproductive and Respiratory Syndrome Virus Horsens Strain on the Reproductive Performance in Pregnant Sows

This study assessed the impact of a PRRSV (porcine reproductive and respiratory syndrome virus) recombinant strain (Horsens strain) on the reproductive performance of na&iuml;ve pregnant sows in the last third of gestation. Fifteen sows were included: four negative reproductive controls (NTX), five infected with a PRRSV-1 field strain (Olot/91, T01), and six infected with the recombinant PRRSV-1 strain (Horsens strain, T02). Piglets were monitored until weaning. Reproductive performance was the primary variable. In sows, viremia and nasal shedding (T01 and T02 groups), and, in piglets, viral load in blood and in lungs, as well as macroscopic lung lesions (T01 and T02 groups), were the secondary variables. The reproductive performance results were numerically different between the two challenged groups. Moreover, viral loads in blood were 1.83 &times; 106 &plusmn; 9.05 &times; 106 copies/mL at farrowing, 1.05 &times; 107 &plusmn; 2.21 &times; 107 copies/mL at weaning from piglets born from T01 animals and 1.64 &times; 103 &plusmn; 7.62 &times; 103 copies/mL at farrowing, 1.95 &times; 103 &plusmn; 1.17 &times; 104 copies/mL at weaning from piglets born from T02 sows. Overall, 68.8% of T01 piglets and 38.1% of T02 piglets presented mild lung lesions. In conclusion, the results suggest that Horsens strain is less virulent than the field strain Olot/91 under these experimental conditions

Effect of metritis on endometrium tissue transcriptome during puerperium in Holstein lactating cows

The objective of this prospective cohort study was to evaluate the effect of parity and uterine healthstatus postpartum on the gene expression profile of the endometrium early post-partum. Twenty-fourHolstein cows were randomly selected (16 multiparous (MP) and 8 primiparous (PP)) and endometriumbiopsies were collected on days 1, 3, and 6 after calving and clinically monitored for metritis. Rectaltemperature was measured twice and fever was defined as a temperature 39.5 C. A case of metritiswas diagnosed with the presence of red-brown watery, foul-smelling uterine discharge or a purulentdischarge with more than 50% pus and fever between days 1 and 6 postpartum. Cows were thenretrospectively selected (cows diagnosed with metritis were paired with healthy ones) to analyze theexpression of 66 genes measured on the NanoString nCounter Analysis System. The genes selected wererelated with adhesion, immune system, steroid and prostaglandin biosynthesis regulation, insulinmetabolism and transcription factors, and nutrient transporters. The results indicated a different patternon genes related to immune function by parity.PTX3,involved in antigen presentation, was increased inhealthy MP compared with healthy PP whereas inflammatory cytokineTNFaand complement-relatedproteinSERPING1was upregulated in MP compared with PP (P<0.05). As expected, presence of ametritis condition affected the expression of genes related to immune function. There was an increasedexpression of the antiviral factorMX2andMYH10gene, which is involved in macrophages recruitment, inmetritic compared with healthy cows (P<0.05). Differences in uterine involution from cows diagnosedwith metritis were reflected by the downregulation ofIGF1(P<0.10), involved in endometriumremodeling, and a possible compensatory upregulation of its receptorIGFR1(P<0.05). A greaterexpression of prostaglandins and oxytocin receptors (PGRandOXTR), involved in the involution process,were observed in metritic PP compared with healthy PP (P<0.05). Overall, it seems that metritissignificantly modulate processes closely tied with the physical involution of the uterus early post-partum(IGF1, IGFR1, PGR, OXTR), whereas both metritis and multiparous cows tended to upregulate genes relatedto immune response (PTX3, TNFa, SERPING1, MX2, MYH10).info:eu-repo/semantics/acceptedVersio

Pre-calving intravaginal administration of lactic acid bacteria reduces metritis prevalence and regulates blood neutrophil gene expression after calving in dairy cattle

Metritis affects up to 40% of dairy cows and it is usually treated with antibiotics. In spite of their advantages, there is an increased concern about antibiotic resistance leading to the research of alternative methods. The aim of this study was to evaluate the effects of a combination of lactic acid bacteria (LAB) on the prevalence of metritis and modulation of endometrial and neutrophil inflammatory markers in dairy cows. One hundred and thirty-five cows were enrolled 3 week before calving and randomly assigned to three treatments. Treatment groups were: (1) two intravaginal doses of LAB/wk during 3 week pre-calving (vaginal, n = 45); (2) an intra-uterine dose, once 1 d after calving (uterine, n = 44); and (3) no intervention (CTRL, n = 45). Metritis was defined as body temperature > 39.5◦C and purulent vaginal discharge (> 50% pus), and diagnosed 6 d after calving. Blood samples were taken at d −14, −10, −7, −4, +1, +3, +6, and +14 relative to calving for non-esterified fatty acids (NEFA) analysis. At d −10, +1, +3, and +6 neutrophils were isolated from blood for gene expression analysis by RT-qPCR. Endometrium biopsies were taken from 30 cows, 15 from CTRL and 15 from the uterine group at d +1, +3, and +6 after calving for pro-inflammatory markers analysis by NanoString®. Vaginal treatment reduced metritis prevalence (6/45) up to 58% compared with CTRL group (14/45), but there was no difference between the uterine and CTRL group. Uterine and vaginal treatments reduced blood neutrophil gene expression. Expression of pro-inflammatory markers in the endometrium did not differ between uterine and CTRL cows. Metritic cows expressed more C-X-C motif chemokine ligand 8 (CXCL8) and interleukin 1 beta (IL1B) at d 3 than healthy cows, whereas healthy cows expressed more CXCL8 at d 1 relative to calving in the endometrium. This study shows a promising potential of LAB probiotics as a preventive treatment against metritis in dairy cows.info:eu-repo/semantics/publishedVersio

Preclinical scenario of targeting myocardial fibrosis with chimeric antigen receptor (CAR) immunotherapy

Fibrosis is present in an important proportion of myocardial disorders. Injury activates cardiac fibroblasts, which deposit excess extracellular matrix, increasing tissue stiffness, impairing cardiac function, and leading to heart failure. Clinical therapies that directly target excessive fibrosis are limited, and more effective treatments are needed. Immunotherapy based on chimeric antigen receptor (CAR) T cells is a novel technique that redirects T lymphocytes toward specific antigens to eliminate the target cells. It is currently used in haematological cancers but has demonstrated efficacy in mouse models of hypertensive cardiac fibrosis, with activated fibroblasts as the target cells. CAR T cell therapy is associated with significant toxicities, but CAR natural killer cells can overcome efficacy and safety limitations. The use of CAR immunotherapy offers a potential alternative to current therapies for fibrosis reduction and restoration of cardiac function in patients with myocardial fibrosis

Pre-calving Intravaginal Administration of Lactic Acid Bacteria Reduces Metritis Prevalence and Regulates Blood Neutrophil Gene Expression After Calving in Dairy Cattle

Metritis affects up to 40% of dairy cows and it is usually treated with antibiotics. In spite of their advantages, there is an increased concern about antibiotic resistance leading to the research of alternative methods. The aim of this study was to evaluate the effects of a combination of lactic acid bacteria (LAB) on the prevalence of metritis and modulation of endometrial and neutrophil inflammatory markers in dairy cows. One hundred and thirty-five cows were enrolled 3 week before calving and randomly assigned to three treatments. Treatment groups were: (1) two intravaginal doses of LAB/wk during 3 week pre-calving (vaginal, n = 45); (2) an intra-uterine dose, once 1 d after calving (uterine, n = 44); and (3) no intervention (CTRL, n = 45). Metritis was defined as body temperature &gt; 39.5°C and purulent vaginal discharge (&gt; 50% pus), and diagnosed 6 d after calving. Blood samples were taken at d −14, −10, −7, −4, +1, +3, +6, and +14 relative to calving for non-esterified fatty acids (NEFA) analysis. At d −10, +1, +3, and +6 neutrophils were isolated from blood for gene expression analysis by RT-qPCR. Endometrium biopsies were taken from 30 cows, 15 from CTRL and 15 from the uterine group at d +1, +3, and +6 after calving for pro-inflammatory markers analysis by NanoString®. Vaginal treatment reduced metritis prevalence (6/45) up to 58% compared with CTRL group (14/45), but there was no difference between the uterine and CTRL group. Uterine and vaginal treatments reduced blood neutrophil gene expression. Expression of pro-inflammatory markers in the endometrium did not differ between uterine and CTRL cows. Metritic cows expressed more C-X-C motif chemokine ligand 8 (CXCL8) and interleukin 1 beta (IL1B) at d 3 than healthy cows, whereas healthy cows expressed more CXCL8 at d 1 relative to calving in the endometrium. This study shows a promising potential of LAB probiotics as a preventive treatment against metritis in dairy cows

Low-density lipoprotein receptor-related protein 1 deficiency in cardiomyocytes reduces susceptibility to insulin resistance and obesity

Background: Low-density lipoprotein receptor-related protein 1 (LRP1) plays a key role in fatty acid metabolism and glucose homeostasis. In the context of dyslipemia, LRP1 is upregulated in the heart. Our aim was to evaluate the impact of cardiomyocyte LRP1 deficiency on high fat diet (HFD)-induced cardiac and metabolic alterations, and to explore the potential mechanisms involved. Methods: We used TnT-iCre transgenic mice with thoroughly tested suitability to delete genes exclusively in cardiomyocytes to generate an experimental mouse model with conditional Lrp1 deficiency in cardiomyocytes (TNT-iCre+-LRP1flox/flox). Findings: Mice with Lrp1-deficient cardiomyocytes (cm-Lrp1-/-) have a normal cardiac function combined with a favorable metabolic phenotype against HFD-induced glucose intolerance and obesity. Glucose intolerance protection was linked to higher hepatic fatty acid oxidation (FAO), lower liver steatosis and increased whole-body energy expenditure. Proteomic studies of the heart revealed decreased levels of cardiac pro-atrial natriuretic peptide (pro-ANP), which was parallel to higher ANP circulating levels. cm-Lrp1-/- mice showed ANP signaling activation that was linked to increased fatty acid (FA) uptake and increased AMPK/ ACC phosphorylation in the liver. Natriuretic peptide receptor A (NPR-A) antagonist completely abolished ANP signaling and metabolic protection in cm-Lrp1-/- mice. Conclusions: These results indicate that an ANP-dependent axis controlled by cardiac LRP1 levels modulates AMPK activity in the liver, energy homeostasis and whole-body metabolism

Use of lactic acid bacteria as a preventive strategy against metritis in dairy cows

Aproximadament el 40% de les vaques lleteres desenvolupen una malaltia uterina durant el post-part que acaba provocant infertilitat. Alguns estudis indiquen que la infecció uterina, causada principalment per Escherichia coli durant la primera setmana post-part, està associada amb la metritis, caracteritzada per la inflamació de l’úter on la vaca és incapaç d’eliminar els bacteris patògens. El tractament antibacterià tradicional que s’utilitza per contrarestar la metritis pot no ser efectiu en tots els casos, sobretot quan hi ha una inflamació prolongada. El primer estudi està enfocat en avaluar l’efecte de 4 possibles probiòtics del grup de bacteris de l’àcid làctic (BAL): Lactobacillus rhamnosus, Pediococcus acidilactici, Lactobacillus sakei i Lactobacillus reuteri, en un cultiu primari d’endometri amb infecció bacteriana i/o inflamació. Els principals resultats obtinguts van ser que P. acidilactici disminuïa la infecció d’E. coli, que L. rhamnosus reduïa significativament la inflamació cel·lular i que L. reuteri era capaç de disminuir la infecció d’E. coli quan les cèl·lules epitelials estaven prèviament inflamades. Durant el segon estudi es van provar 4 combinacions diferents de BAL a partir dels resultats obtinguts en el primer estudi. Es va avaluar la capacitat d’aquestes combinacions de disminuir la infecció d’E. coli i reduir la inflamació en el mateix cultiu primari. La combinació composta per L. rhamnous/ P. acidilactici/ L. reuteri amb una ràtio de 12/12/1 va ser la seleccionada. Llavors es va decidir comprovar que aquesta combinació continués sent eficaç en un model ex vivo (explants d’endometri). Els resultats obtinguts confirmaven la capacitat de la combinació de BAL de reduir la inflamació tissular. Per altra banda els assajos amb microscòpia electrònica mostraven un efecte protector de BAL en les cèl·lules epitelials. Hi havia menys necrosis, menys dany mitocondrial i més moc en les cèl·lules tractades amb BAL que en les no tractades. En el tercer estudi es va aplicar la combinació de BAL intravaginalment a vaques i al cap de 3 setmanes es va recol·lectar els seus endometris per obtenir-ne explants i infectar-los amb E. coli ex vivo. No es van observar diferències en els marcadors d’inflamació entre les vaques tractades amb BAL o les vaques control, ni en el número de Lactobacillus que hi havia a l’endometri de les vaques. Per altra banda, les vaques tractades amb BAL tendien a tenir menys presència d’E. coli a la vagina que les vaques control, i a més, expressaven menys B-defensins i MUC1, considerats marcadors d’infecció. Finalment, en el quart estudi, es van analitzar els efectes in vivo de la combinació de BAL sobre la incidència de metritis i inflamació de l’endometri en vaques de llet quan s’administrava intravaginalment durant 3 setmanes pre-part i o a l’úter un dia post-part. Els principals resultats obtinguts van ser que el tractament vaginal reduïa un 58% la prevalença de metritis comparada amb el grup control mentre que el tractament endometrial no la variava. No es va observar cap diferència amb els marcadors d’inflamació però els dos tractaments disminuïen l’activitatneutrofílica.Approximately 40% of dairy cows develop a uterine disease during the post-partum leading to infertility. Several studies indicate that uterine infection, mainly caused by Escherichia coli during the first week post-partum, is associated with metritis, characterized by inflammation in the endometrium where the cow is not able to clear pathogenic bacteria. The traditional antimicrobial treatment may lack efficacy, especially in cases of sustained inflammation. The first study is focused in the evaluation of 4 possible probiotics belonging to the lactic acid bacteria (LAB): Lactobacillus rhamnosus, Pediococcus acidilactici, Lactobacillus sakei, and Lactobacillus reuteri, in an endometrial primary culture against bacterial infection and inflammation. The main results were that P. acidilactici was able to reduce E. coli infection, L. rhamnosus diminished cellular inflammation, and L. reuteri reduced E. coli infection when the epithelial cells were inflammated. On the second study, 4 different LAB combinations based on the results of the first study, were tested using the same primary culture. The combination composed by L. rhamnosus/ P. acidilactici/ L. reuteri with a ratio of 12/12/1 was selected. Then, this combination was tested in an ex vivo model (endometrial explants). The obtained results confirmed the capacity of this LAB combination to reduce tissular inflammation. On the other hand, electron microscopy assays showed a protective effect of LAB in endometrial epithelial cells. There was less necrosis, mitochondrial damage, and more mucus in the surface of LAB-treated cells than not-treated cells. In the third study, LAB combination was applied in vivo in the vagina of several cows, and 3 weeks later, the endometrium of those animals were collected. Explants were made from the endometrium and then infected with E. coli. No differences were observed in the inflammation markers between LAB-treated and control cows, or in the final quantification of Lactobacillus in the endometrium. On the other hand, LAB-treated cows tended to have less presence of E. coli in the vagina than control cows and, moreover, they expressed less B-defensins and MUC1, considerate markers of infection. Finally, on the fourth study, the effects of LAB combination were analyzed in vivo quantifying metritis prevalence and endometrial inflammation in dairy cows when the LAB combination was applied intravaginally during 3 weeks pre-partum or intra-uterine, 1 day after calving. The main results were that the vaginal treatment reduced metritis prevalence up to 58% compared with the control cows while no differences were observed with the endometrial treatment. No differences were found in the inflammation markers whereas both treatments (vaginal and endometrial) were able to modulate neutrophilic activit