Abstract The aim of this study was to evaluate the effect of feeding method on eating and sorting... more Abstract The aim of this study was to evaluate the effect of feeding method on eating and sorting behavior, rumen acidosis, and apparent total tract digestibility of crossbred Angus bulls fed a high-concentrate diet. Twenty-one Angus beef bulls (497 ± 7.7 kg of initial BW, and 324 ± 3.0 d of age) were housed individually and fed ad libitum. Three experimental treatments were tested: complete feed of pellet and chopped straw in a single feeder (TMR), pellet and chopped straw fed separately in two feeders (SS); pellet and long unprocessed straw fed separately in two feeders (LS). Feed consumption, fecal and bloat scoring were recorded daily. Every 2 wk TMR sorting, eating behavior, general activity, social and oral behaviors, and BW were recorded. At days 14 and 35 apparent total tract digestibility during one week was measured. At days 28 and 42 rumen samples were collected via rumenocentesis to measure rumen pH and determine ruminal volatile fatty acids (VFA) concentrations. At the study end (day 57) animals at slaughterhouse liver and rumen wall color and lesions were recorded by a macroscopic inspection. No differences among treatments in total DMI, and fecal and bloat scoring were observed. The straw to concentrate ratio was smaller in the SS and LS treatments (8–92) than in the TMR (15–85), and sorting analyses indicated that TMR bulls refused large particles (> 4 mm) and small particles (
Dietary enzyme supplementation, as a feed additive, has been well adopted in monogastric producti... more Dietary enzyme supplementation, as a feed additive, has been well adopted in monogastric production to increase feed efficiency. However, in ruminants, considerably fewer studies have been done and fewer, if any, commercial enzymes have been adopted as a feeding strategy. Feedlot cattle are commonly fed high-starch diets, with varying starch digestibilities depending on type of grain, and degree of grain processing. Improvement in starch digestibility in low processed grain diets will undoubtedly warrant economic benefits to feedlot producers and reduce environmental impact of intensive beef production. In this study, we have shown that dietary glucoamylase supplementation improved 7 to 13% apparent digestibility of dry matter and starch in bulls fed rolled corn-based diets, suggesting that enzyme (glucoamylase) supplementation could be a promising strategy to improve starch efficiency for finishing beef cattle.
Uterine function in cattle is compromised by bacterial contamination and inflammation after calvi... more Uterine function in cattle is compromised by bacterial contamination and inflammation after calving. The objective of this study was to select a combination of lactic acid bacteria (LAB) to decrease endometrium inflammation and Escherichia coli infection. Primary endometrial epithelial cells were cultured in vitro to select the most favorable LAB combination modulating basal tissue inflammation and E. coli infection. Supernatants were obtained to determine expression of pro-inflammatory cytokines, and E. coli infection was evaluated after harvesting the tissue and plate counting. The selected LAB combination was tested in uterus explants to assess its capacity to modulate basal and acute inflammation (associated with E. coli infection). The combination of Lactobacillus rhamnosus, Pediococcus acidilactici, and Lactobacillus reuteri at a ratio of 25:25:2, respectively, reduced E. coli infection in vitro with (89.77%) or without basal tissue inflammation (95.10%) compared with single LAB strains. Lactic acid bacteria treatment reduced CXCL8 and IL1B expression 4.7-and 2.2-fold, respectively, under acute inflammation. Ex vivo, the tested LAB combination reduced acute inflammation under E. coli infection, decreasing IL-8, IL-1β, and IL-6 up to 2.2-, 2.5-, and 2.2-fold, respectively. In the total inflammation model, the LAB combination decreased IL-8 1.6-fold and IL-6 1.2-fold. Ultrastructural evaluation of the tissue suggested no direct interaction between the LAB and E. coli, although pathological effects of E. coli in endometrial cells were greatly diminished or even reversed by the LAB combination. This study shows the promising potential of LAB probiotics for therapeutic use against endometrial inflammation and infection.
The objective of this prospective cohort study was to evaluate the effect of parity and uterine h... more The objective of this prospective cohort study was to evaluate the effect of parity and uterine health status postpartum on the gene expression profile of the endometrium early post-partum. Twenty-four Holstein cows were randomly selected (16 multiparous (MP) and 8 primiparous (PP)) and endometrium biopsies were collected on days 1, 3, and 6 after calving and clinically monitored for metritis. Rectal temperature was measured twice and fever was defined as a temperature ≥ 39.5°C. A case of metritis was diagnosed with the presence of red-brown watery, foul-smelling uterine discharge or a purulent discharge with more than 50% pus and fever between days 1 and 6 postpartum. Cows were then retrospectively selected (cows diagnosed with metritis were paired with healthy ones) to analyze the expression of 66 genes measured on the NanoString nCounter Analysis System. The genes selected were related with adhesion, immune system, steroid and prostaglandin biosynthesis regulation, insulin metabolism and transcription factors, and nutrient transporters. The results indicated a different pattern on genes related to immune function by parity. PTX3, involved in antigen presentation, was increased in healthy MP compared with healthy PP whereas inflammatory cytokine TNFα and complement-related protein SERPING1 was upregulated in MP compared Revised
Twenty-one crossbred Angus bulls (497 ± 7.7 kg of BW and 324 ± 3.0 d of age) were housed individu... more Twenty-one crossbred Angus bulls (497 ± 7.7 kg of BW and 324 ± 3.0 d of age) were housed individually to evaluate the effect feeding method on feed intake, sorting, animal behavior, and digestibility. Dietary treatments, offered ad libitum, were: complete feed of pellet (concentrate) mixed with chopped straw (5 cm length; TMR); pellet and chopped straw (5 cm length; SS); pellet and long unprocessed straw (15–20 cm length; LS). In SS and LS straw and concentrate were fed separate feeders. Feed intake, fecal and bloat scoring were recorded daily. Behavior (scan sampling), BW, and TMR particle size sorting were recorded fortnightly. Total tract apparent digestibility (day 14 and 35) and rumen pH and VFA samples via rumenocentesis (day 28 and 42) were analyzed. Data were analyzed using a mixed-effect model. No differences in concentrate and straw intake, fecal and bloat scoring among treatments were observed. The straw to concentrate ratio did not differ between SS and LS (8 to 92), and was smaller than the ratio fixed in TMR (15 to 85). TMR bulls sorted (P < 0.05) particles with a size between 1.7 and 4 mm. Treatment did not affect rumination time. TMR showed (P < 0.05) more oral non-nutritive behaviors than SS. Rumen pH was greater (P < 0.01) in TMR than in SS and LS, although it was above 5.6 in all treatments. Rumen acetate to propionate ratio was greater (P < 0.05) in TMR than in SS and LS. LS bulls had greater DM (P < 0.05) and CP (P < 0.01) digestibility compared with SS or TMR bulls. Feeding concentrate and straw (short or long) separately does not increase the risk of rumen acidosis compared with TMR, and feeding long straw separately from the concentrate improves the total tract apparent DM and CP digestibility.
low numbers in epithelial cells surface and in no case formed biofilms or interactions between ea... more low numbers in epithelial cells surface and in no case formed biofilms or interactions between each other. Escherichia coli abundance was lower (P < 0.05) in samples treated with LAB than in those infected with E. coli alone. Healthy epithelium was observed in cells treated with LAB (epithelial cells with normal size and shape and normal aspect of microvilli), whereas in cultures infected with E. coli, abundant areas with cell debris and bacilli in epithelial cell surface were observed. The incidence of necrosis (as assessed by TEM) in E. coli samples tended (P = 0.07) to be greater than in noninfected cultures. Control or LAB preincubated cells showed less mitochondrial damage (P = 0.01) than nontreated cells, a parameter strongly related to cell death. Overall, LAB appear to offer protection against E. coli, by mechanism different than the formation of biofilms, and thus, LAB combinations could be used as a preventive strategy for metritis.
This study assessed the impact of a PRRSV (porcine reproductive and respiratory syndrome virus) r... more This study assessed the impact of a PRRSV (porcine reproductive and respiratory syndrome virus) recombinant strain (Horsens strain) on the reproductive performance of naïve 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 × 10 6 ± 9.05 × 10 6 copies/mL at farrowing, 1.05 × 10 7 ± 2.21 × 10 7 copies/mL at weaning from piglets born from T01 animals and 1.64 × 10 3 ± 7.62 × 10 3 copies/mL at farrowing, 1.95 × 10 3 ± 1.17 × 10 4 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.
About 40% of dairy cattle develop uterine disease during postpartum period, causing infertility. ... more About 40% of dairy cattle develop uterine disease during postpartum period, causing infertility. Some studies indicate that uterine infection, predominantly by Escherichia coli in the first week postpartum, is associated with metritis, an uterus inflammation in which the cow fails to completely clear bacterial contaminants. The aim of this study was to evaluate the potential of four lactic acid bacteria (LAB) (Lactobacillus rhamnosus, Pediococcus acidilactici, Lactobacillus reuteri, and Lactobacillus sakei) to modulate the E coli infection and inflammation in endometrial cells. Primary endometrial epithelial cells were isolated from fresh endometrium of a healthy cow and cultured in vitro to evaluate the effects of LAB at three different doses. Cell extracts were obtained to analyze the expression of proinflammatory cytokines and to quantify E coli infection on MacConkey agar plates. L sakei and L reuteri showed a positive effect preventing E coli infection (87% and 78%, respectively, P &amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;lt; 0.001); however, they were also associated to a dose-variable effect on tissular inflammation that could further exacerbate the proinflammatory status. Infection of E coli was clearly reduced (P &amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;lt; 0.001) up to an 83% with P acidilactici, whereas, the expression of proinflammatory cytokines IL-8 and IL-1β dropped significantly (P &amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;lt; 0.001) up to 85.11 and 5.24 folds, respectively, in the presence of L rhamnosus. In conclusion, these results demonstrate a clear potential of some LAB in the modulation of endometrial infection and inflammation in cattle.
Metritis affects up to 40% of dairy cows and it is usually treated with antibiotics. In spite of ... more 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.
The aim of this study was to evaluate the effect of 2 glucoamylases (GA) and the combination of o... more The aim of this study was to evaluate the effect of 2 glucoamylases (GA) and the combination of one GA with a neutral protease on total tract apparent digestibility in 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 housed individually in pens and were distributed in 4 blocks of 4 animals balanced by BW. The experimental design was a 4 x 4 Latin square (4 blocks and 4 periods) with periods of 2 wks. Four treatments were tested; 1) control, a blank solution of 0.2% potassium sorbate and 0.6% sodium benzoate corresponding to the preservatives used in the enzyme preparation, 2) a GA preparation from Trichoderma reesei; 3) a GA preparation from Aspergillus fumigatus; 4) and a GA from Aspergillus fumigatus and a Bacillus amyloliquefaciens neutral protease preparation. These enzyme solutions were daily mixed was mixed in a dry TMR with 70% of rolled corn at a rate of 10 mL for 40 kg TMR. From day ...
Twenty-one crossbred Angus bulls (497 ± 7.7 kg of BW and 324 ± 3.0 d of age) were housed individu... more Twenty-one crossbred Angus bulls (497 ± 7.7 kg of BW and 324 ± 3.0 d of age) were housed individually to evaluate the effect feeding method on feed intake, sorting, animal behavior, and digestibility. Dietary treatments, offered ad libitum, were: complete feed of pellet (concentrate) mixed with chopped straw (5 cm length; TMR); pellet and chopped straw (5 cm length; SS); pellet and long unprocessed straw (15–20 cm length; LS). In SS and LS straw and concentrate were fed separate feeders. Feed intake, fecal and bloat scoring were recorded daily. Behavior (scan sampling), BW, and TMR particle size sorting were recorded fortnightly. Total tract apparent digestibility (day 14 and 35) and rumen pH and VFA samples via rumenocentesis (day 28 and 42) were analyzed. Data were analyzed using a mixed-effect model. No differences in concentrate and straw intake, fecal and bloat scoring among treatments were observed. The straw to concentrate ratio did not differ between SS and LS (8 to 92), and...
The anti-inflammatory phytohormone abscisic acid (ABA) modulates immune and inflammatory response... more The anti-inflammatory phytohormone abscisic acid (ABA) modulates immune and inflammatory responses in mouse models of colitis and obesity. ABA has been identified as a ligand of lanthionine synthetase C-like 2, a novel therapeutic target upstream of the peroxisome proliferatoractivated receptor γ (PPAR γ) pathway. The goal of this study was to investigate the immune modulatory mechanisms underlying the anti-inflammatory efficacy of ABA against influenzaassociated pulmonary inflammation. Wild type (WT) and conditional knockout mice with defective PPAR γ expression in lung epithelial and hematopoietic cells (cKO) treated orally with or without ABA (100 mg/kg diet) were challenged with Influenza A/Udorn (H3N2) to assess ABA's impact in disease, lung lesions and gene expression. Dietary ABA ameliorated disease activity, lung inflammatory pathology, accelerated recovery and increased survival in WT mice. ABA suppressed leukocyte infiltration and MCP-1 mRNA expression in WT mice through PPAR γ, since this effect was abrogated in cKO mice. ABA ameliorated disease when administered therapeutically on the same day of the infection to WT but not mice lacking PPAR γ in myeloid cells. We also show that ABA's greater impact is between days 7 and 10 post-challenge when it regulates the expression of genes involved in resolution, like 5 lipoxygenase and other members of the 5-lipoxygenase pathway. Furthermore, ABA significantly increased the expression of the immunoregulatory cytokine IL-10 in WT mice. Our results show that ABA, given preventively or therapeutically, ameliorates influenza virus-induced pathology by activating PPAR γ in pulmonary immune cells, suppressing initial proinflammatory responses and promoting resolution.
Aproximadament el 40% de les vaques lleteres desenvolupen una malaltia uterina durant el post-par... more Aproximadament el 40% de les vaques lleteres desenvolupen una malaltia uterina durant el post-part que acaba provocant infertilitat. Alguns estudis indiquen que la infeccio uterina, causada principalment per Escherichia coli durant la primera setmana post-part, esta associada amb la metritis, caracteritzada per la inflamacio de l’uter on la vaca es incapac d’eliminar els bacteris patogens. El tractament antibacteria tradicional que s’utilitza per contrarestar la metritis pot no ser efectiu en tots els casos, sobretot quan hi ha una inflamacio prolongada. El primer estudi esta enfocat en avaluar l’efecte de 4 possibles probiotics del grup de bacteris de l’acid lactic (BAL): Lactobacillus rhamnosus, Pediococcus acidilactici, Lactobacillus sakei i Lactobacillus reuteri, en un cultiu primari d’endometri amb infeccio bacteriana i/o inflamacio. Els principals resultats obtinguts van ser que P. acidilactici disminuia la infeccio d’E. coli, que L. rhamnosus reduia significativament la infla...
This study assessed the impact of a PRRSV (porcine reproductive and respiratory syndrome virus) r... more This study assessed the impact of a PRRSV (porcine reproductive and respiratory syndrome virus) recombinant strain (Horsens strain) on the reproductive performance of naïve 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 × 106 ± 9.05 × 106 copies/mL at farrowing, 1.05 × 107 ± 2.21 × 107 copies/mL at weaning from piglets born from T01 animals and 1.64 × 103 ± 7.62 × ...
Senescence has been previously regarded as protective against tumor growth. However, there is evi... more Senescence has been previously regarded as protective against tumor growth. However, there is evidence that senescent cells observed in the tumor stroma may support cancer progression. The presence of senescent stromal cells in non-small cell lung cancer (NSCLC) remains poorly defined. To address this question, we examined senescent associated signatures in primary cultures of human fibroblasts obtained from 3 major NSCLC subtypes (adenocarcinoma/ADC, squamous cell carcinoma/SCC, large cell carcinoma/LCC) or paired controls. The senescence-associated beta-galactosidase (beta-gal) assay was negative in >90% of control fibroblasts (11/12). Likewise, tumor-assocaited fibroblasts (TAFs) from either ADC or SCC were negative for beta-gal assay in 80% of patients (4/5), whereas <10% were beta-gal positive in the remaining 20% of patients. In contrast, beta-gal positive TAFs were observed in 100% of LCC patients (3/3), with an average of ∼30% positive TAFs. In agreement with these fin...
Abstract The aim of this study was to evaluate the effect of feeding method on eating and sorting... more Abstract The aim of this study was to evaluate the effect of feeding method on eating and sorting behavior, rumen acidosis, and apparent total tract digestibility of crossbred Angus bulls fed a high-concentrate diet. Twenty-one Angus beef bulls (497 ± 7.7 kg of initial BW, and 324 ± 3.0 d of age) were housed individually and fed ad libitum. Three experimental treatments were tested: complete feed of pellet and chopped straw in a single feeder (TMR), pellet and chopped straw fed separately in two feeders (SS); pellet and long unprocessed straw fed separately in two feeders (LS). Feed consumption, fecal and bloat scoring were recorded daily. Every 2 wk TMR sorting, eating behavior, general activity, social and oral behaviors, and BW were recorded. At days 14 and 35 apparent total tract digestibility during one week was measured. At days 28 and 42 rumen samples were collected via rumenocentesis to measure rumen pH and determine ruminal volatile fatty acids (VFA) concentrations. At the study end (day 57) animals at slaughterhouse liver and rumen wall color and lesions were recorded by a macroscopic inspection. No differences among treatments in total DMI, and fecal and bloat scoring were observed. The straw to concentrate ratio was smaller in the SS and LS treatments (8–92) than in the TMR (15–85), and sorting analyses indicated that TMR bulls refused large particles (> 4 mm) and small particles (
Abstract The aim of this study was to evaluate the effect of feeding method on eating and sorting... more Abstract The aim of this study was to evaluate the effect of feeding method on eating and sorting behavior, rumen acidosis, and apparent total tract digestibility of crossbred Angus bulls fed a high-concentrate diet. Twenty-one Angus beef bulls (497 ± 7.7 kg of initial BW, and 324 ± 3.0 d of age) were housed individually and fed ad libitum. Three experimental treatments were tested: complete feed of pellet and chopped straw in a single feeder (TMR), pellet and chopped straw fed separately in two feeders (SS); pellet and long unprocessed straw fed separately in two feeders (LS). Feed consumption, fecal and bloat scoring were recorded daily. Every 2 wk TMR sorting, eating behavior, general activity, social and oral behaviors, and BW were recorded. At days 14 and 35 apparent total tract digestibility during one week was measured. At days 28 and 42 rumen samples were collected via rumenocentesis to measure rumen pH and determine ruminal volatile fatty acids (VFA) concentrations. At the study end (day 57) animals at slaughterhouse liver and rumen wall color and lesions were recorded by a macroscopic inspection. No differences among treatments in total DMI, and fecal and bloat scoring were observed. The straw to concentrate ratio was smaller in the SS and LS treatments (8–92) than in the TMR (15–85), and sorting analyses indicated that TMR bulls refused large particles (> 4 mm) and small particles (
Dietary enzyme supplementation, as a feed additive, has been well adopted in monogastric producti... more Dietary enzyme supplementation, as a feed additive, has been well adopted in monogastric production to increase feed efficiency. However, in ruminants, considerably fewer studies have been done and fewer, if any, commercial enzymes have been adopted as a feeding strategy. Feedlot cattle are commonly fed high-starch diets, with varying starch digestibilities depending on type of grain, and degree of grain processing. Improvement in starch digestibility in low processed grain diets will undoubtedly warrant economic benefits to feedlot producers and reduce environmental impact of intensive beef production. In this study, we have shown that dietary glucoamylase supplementation improved 7 to 13% apparent digestibility of dry matter and starch in bulls fed rolled corn-based diets, suggesting that enzyme (glucoamylase) supplementation could be a promising strategy to improve starch efficiency for finishing beef cattle.
Uterine function in cattle is compromised by bacterial contamination and inflammation after calvi... more Uterine function in cattle is compromised by bacterial contamination and inflammation after calving. The objective of this study was to select a combination of lactic acid bacteria (LAB) to decrease endometrium inflammation and Escherichia coli infection. Primary endometrial epithelial cells were cultured in vitro to select the most favorable LAB combination modulating basal tissue inflammation and E. coli infection. Supernatants were obtained to determine expression of pro-inflammatory cytokines, and E. coli infection was evaluated after harvesting the tissue and plate counting. The selected LAB combination was tested in uterus explants to assess its capacity to modulate basal and acute inflammation (associated with E. coli infection). The combination of Lactobacillus rhamnosus, Pediococcus acidilactici, and Lactobacillus reuteri at a ratio of 25:25:2, respectively, reduced E. coli infection in vitro with (89.77%) or without basal tissue inflammation (95.10%) compared with single LAB strains. Lactic acid bacteria treatment reduced CXCL8 and IL1B expression 4.7-and 2.2-fold, respectively, under acute inflammation. Ex vivo, the tested LAB combination reduced acute inflammation under E. coli infection, decreasing IL-8, IL-1β, and IL-6 up to 2.2-, 2.5-, and 2.2-fold, respectively. In the total inflammation model, the LAB combination decreased IL-8 1.6-fold and IL-6 1.2-fold. Ultrastructural evaluation of the tissue suggested no direct interaction between the LAB and E. coli, although pathological effects of E. coli in endometrial cells were greatly diminished or even reversed by the LAB combination. This study shows the promising potential of LAB probiotics for therapeutic use against endometrial inflammation and infection.
The objective of this prospective cohort study was to evaluate the effect of parity and uterine h... more The objective of this prospective cohort study was to evaluate the effect of parity and uterine health status postpartum on the gene expression profile of the endometrium early post-partum. Twenty-four Holstein cows were randomly selected (16 multiparous (MP) and 8 primiparous (PP)) and endometrium biopsies were collected on days 1, 3, and 6 after calving and clinically monitored for metritis. Rectal temperature was measured twice and fever was defined as a temperature ≥ 39.5°C. A case of metritis was diagnosed with the presence of red-brown watery, foul-smelling uterine discharge or a purulent discharge with more than 50% pus and fever between days 1 and 6 postpartum. Cows were then retrospectively selected (cows diagnosed with metritis were paired with healthy ones) to analyze the expression of 66 genes measured on the NanoString nCounter Analysis System. The genes selected were related with adhesion, immune system, steroid and prostaglandin biosynthesis regulation, insulin metabolism and transcription factors, and nutrient transporters. The results indicated a different pattern on genes related to immune function by parity. PTX3, involved in antigen presentation, was increased in healthy MP compared with healthy PP whereas inflammatory cytokine TNFα and complement-related protein SERPING1 was upregulated in MP compared Revised
Twenty-one crossbred Angus bulls (497 ± 7.7 kg of BW and 324 ± 3.0 d of age) were housed individu... more Twenty-one crossbred Angus bulls (497 ± 7.7 kg of BW and 324 ± 3.0 d of age) were housed individually to evaluate the effect feeding method on feed intake, sorting, animal behavior, and digestibility. Dietary treatments, offered ad libitum, were: complete feed of pellet (concentrate) mixed with chopped straw (5 cm length; TMR); pellet and chopped straw (5 cm length; SS); pellet and long unprocessed straw (15–20 cm length; LS). In SS and LS straw and concentrate were fed separate feeders. Feed intake, fecal and bloat scoring were recorded daily. Behavior (scan sampling), BW, and TMR particle size sorting were recorded fortnightly. Total tract apparent digestibility (day 14 and 35) and rumen pH and VFA samples via rumenocentesis (day 28 and 42) were analyzed. Data were analyzed using a mixed-effect model. No differences in concentrate and straw intake, fecal and bloat scoring among treatments were observed. The straw to concentrate ratio did not differ between SS and LS (8 to 92), and was smaller than the ratio fixed in TMR (15 to 85). TMR bulls sorted (P < 0.05) particles with a size between 1.7 and 4 mm. Treatment did not affect rumination time. TMR showed (P < 0.05) more oral non-nutritive behaviors than SS. Rumen pH was greater (P < 0.01) in TMR than in SS and LS, although it was above 5.6 in all treatments. Rumen acetate to propionate ratio was greater (P < 0.05) in TMR than in SS and LS. LS bulls had greater DM (P < 0.05) and CP (P < 0.01) digestibility compared with SS or TMR bulls. Feeding concentrate and straw (short or long) separately does not increase the risk of rumen acidosis compared with TMR, and feeding long straw separately from the concentrate improves the total tract apparent DM and CP digestibility.
low numbers in epithelial cells surface and in no case formed biofilms or interactions between ea... more low numbers in epithelial cells surface and in no case formed biofilms or interactions between each other. Escherichia coli abundance was lower (P < 0.05) in samples treated with LAB than in those infected with E. coli alone. Healthy epithelium was observed in cells treated with LAB (epithelial cells with normal size and shape and normal aspect of microvilli), whereas in cultures infected with E. coli, abundant areas with cell debris and bacilli in epithelial cell surface were observed. The incidence of necrosis (as assessed by TEM) in E. coli samples tended (P = 0.07) to be greater than in noninfected cultures. Control or LAB preincubated cells showed less mitochondrial damage (P = 0.01) than nontreated cells, a parameter strongly related to cell death. Overall, LAB appear to offer protection against E. coli, by mechanism different than the formation of biofilms, and thus, LAB combinations could be used as a preventive strategy for metritis.
This study assessed the impact of a PRRSV (porcine reproductive and respiratory syndrome virus) r... more This study assessed the impact of a PRRSV (porcine reproductive and respiratory syndrome virus) recombinant strain (Horsens strain) on the reproductive performance of naïve 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 × 10 6 ± 9.05 × 10 6 copies/mL at farrowing, 1.05 × 10 7 ± 2.21 × 10 7 copies/mL at weaning from piglets born from T01 animals and 1.64 × 10 3 ± 7.62 × 10 3 copies/mL at farrowing, 1.95 × 10 3 ± 1.17 × 10 4 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.
About 40% of dairy cattle develop uterine disease during postpartum period, causing infertility. ... more About 40% of dairy cattle develop uterine disease during postpartum period, causing infertility. Some studies indicate that uterine infection, predominantly by Escherichia coli in the first week postpartum, is associated with metritis, an uterus inflammation in which the cow fails to completely clear bacterial contaminants. The aim of this study was to evaluate the potential of four lactic acid bacteria (LAB) (Lactobacillus rhamnosus, Pediococcus acidilactici, Lactobacillus reuteri, and Lactobacillus sakei) to modulate the E coli infection and inflammation in endometrial cells. Primary endometrial epithelial cells were isolated from fresh endometrium of a healthy cow and cultured in vitro to evaluate the effects of LAB at three different doses. Cell extracts were obtained to analyze the expression of proinflammatory cytokines and to quantify E coli infection on MacConkey agar plates. L sakei and L reuteri showed a positive effect preventing E coli infection (87% and 78%, respectively, P &amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;lt; 0.001); however, they were also associated to a dose-variable effect on tissular inflammation that could further exacerbate the proinflammatory status. Infection of E coli was clearly reduced (P &amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;lt; 0.001) up to an 83% with P acidilactici, whereas, the expression of proinflammatory cytokines IL-8 and IL-1β dropped significantly (P &amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;lt; 0.001) up to 85.11 and 5.24 folds, respectively, in the presence of L rhamnosus. In conclusion, these results demonstrate a clear potential of some LAB in the modulation of endometrial infection and inflammation in cattle.
Metritis affects up to 40% of dairy cows and it is usually treated with antibiotics. In spite of ... more 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.
The aim of this study was to evaluate the effect of 2 glucoamylases (GA) and the combination of o... more The aim of this study was to evaluate the effect of 2 glucoamylases (GA) and the combination of one GA with a neutral protease on total tract apparent digestibility in 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 housed individually in pens and were distributed in 4 blocks of 4 animals balanced by BW. The experimental design was a 4 x 4 Latin square (4 blocks and 4 periods) with periods of 2 wks. Four treatments were tested; 1) control, a blank solution of 0.2% potassium sorbate and 0.6% sodium benzoate corresponding to the preservatives used in the enzyme preparation, 2) a GA preparation from Trichoderma reesei; 3) a GA preparation from Aspergillus fumigatus; 4) and a GA from Aspergillus fumigatus and a Bacillus amyloliquefaciens neutral protease preparation. These enzyme solutions were daily mixed was mixed in a dry TMR with 70% of rolled corn at a rate of 10 mL for 40 kg TMR. From day ...
Twenty-one crossbred Angus bulls (497 ± 7.7 kg of BW and 324 ± 3.0 d of age) were housed individu... more Twenty-one crossbred Angus bulls (497 ± 7.7 kg of BW and 324 ± 3.0 d of age) were housed individually to evaluate the effect feeding method on feed intake, sorting, animal behavior, and digestibility. Dietary treatments, offered ad libitum, were: complete feed of pellet (concentrate) mixed with chopped straw (5 cm length; TMR); pellet and chopped straw (5 cm length; SS); pellet and long unprocessed straw (15–20 cm length; LS). In SS and LS straw and concentrate were fed separate feeders. Feed intake, fecal and bloat scoring were recorded daily. Behavior (scan sampling), BW, and TMR particle size sorting were recorded fortnightly. Total tract apparent digestibility (day 14 and 35) and rumen pH and VFA samples via rumenocentesis (day 28 and 42) were analyzed. Data were analyzed using a mixed-effect model. No differences in concentrate and straw intake, fecal and bloat scoring among treatments were observed. The straw to concentrate ratio did not differ between SS and LS (8 to 92), and...
The anti-inflammatory phytohormone abscisic acid (ABA) modulates immune and inflammatory response... more The anti-inflammatory phytohormone abscisic acid (ABA) modulates immune and inflammatory responses in mouse models of colitis and obesity. ABA has been identified as a ligand of lanthionine synthetase C-like 2, a novel therapeutic target upstream of the peroxisome proliferatoractivated receptor γ (PPAR γ) pathway. The goal of this study was to investigate the immune modulatory mechanisms underlying the anti-inflammatory efficacy of ABA against influenzaassociated pulmonary inflammation. Wild type (WT) and conditional knockout mice with defective PPAR γ expression in lung epithelial and hematopoietic cells (cKO) treated orally with or without ABA (100 mg/kg diet) were challenged with Influenza A/Udorn (H3N2) to assess ABA's impact in disease, lung lesions and gene expression. Dietary ABA ameliorated disease activity, lung inflammatory pathology, accelerated recovery and increased survival in WT mice. ABA suppressed leukocyte infiltration and MCP-1 mRNA expression in WT mice through PPAR γ, since this effect was abrogated in cKO mice. ABA ameliorated disease when administered therapeutically on the same day of the infection to WT but not mice lacking PPAR γ in myeloid cells. We also show that ABA's greater impact is between days 7 and 10 post-challenge when it regulates the expression of genes involved in resolution, like 5 lipoxygenase and other members of the 5-lipoxygenase pathway. Furthermore, ABA significantly increased the expression of the immunoregulatory cytokine IL-10 in WT mice. Our results show that ABA, given preventively or therapeutically, ameliorates influenza virus-induced pathology by activating PPAR γ in pulmonary immune cells, suppressing initial proinflammatory responses and promoting resolution.
Aproximadament el 40% de les vaques lleteres desenvolupen una malaltia uterina durant el post-par... more Aproximadament el 40% de les vaques lleteres desenvolupen una malaltia uterina durant el post-part que acaba provocant infertilitat. Alguns estudis indiquen que la infeccio uterina, causada principalment per Escherichia coli durant la primera setmana post-part, esta associada amb la metritis, caracteritzada per la inflamacio de l’uter on la vaca es incapac d’eliminar els bacteris patogens. El tractament antibacteria tradicional que s’utilitza per contrarestar la metritis pot no ser efectiu en tots els casos, sobretot quan hi ha una inflamacio prolongada. El primer estudi esta enfocat en avaluar l’efecte de 4 possibles probiotics del grup de bacteris de l’acid lactic (BAL): Lactobacillus rhamnosus, Pediococcus acidilactici, Lactobacillus sakei i Lactobacillus reuteri, en un cultiu primari d’endometri amb infeccio bacteriana i/o inflamacio. Els principals resultats obtinguts van ser que P. acidilactici disminuia la infeccio d’E. coli, que L. rhamnosus reduia significativament la infla...
This study assessed the impact of a PRRSV (porcine reproductive and respiratory syndrome virus) r... more This study assessed the impact of a PRRSV (porcine reproductive and respiratory syndrome virus) recombinant strain (Horsens strain) on the reproductive performance of naïve 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 × 106 ± 9.05 × 106 copies/mL at farrowing, 1.05 × 107 ± 2.21 × 107 copies/mL at weaning from piglets born from T01 animals and 1.64 × 103 ± 7.62 × ...
Senescence has been previously regarded as protective against tumor growth. However, there is evi... more Senescence has been previously regarded as protective against tumor growth. However, there is evidence that senescent cells observed in the tumor stroma may support cancer progression. The presence of senescent stromal cells in non-small cell lung cancer (NSCLC) remains poorly defined. To address this question, we examined senescent associated signatures in primary cultures of human fibroblasts obtained from 3 major NSCLC subtypes (adenocarcinoma/ADC, squamous cell carcinoma/SCC, large cell carcinoma/LCC) or paired controls. The senescence-associated beta-galactosidase (beta-gal) assay was negative in >90% of control fibroblasts (11/12). Likewise, tumor-assocaited fibroblasts (TAFs) from either ADC or SCC were negative for beta-gal assay in 80% of patients (4/5), whereas <10% were beta-gal positive in the remaining 20% of patients. In contrast, beta-gal positive TAFs were observed in 100% of LCC patients (3/3), with an average of ∼30% positive TAFs. In agreement with these fin...
Abstract The aim of this study was to evaluate the effect of feeding method on eating and sorting... more Abstract The aim of this study was to evaluate the effect of feeding method on eating and sorting behavior, rumen acidosis, and apparent total tract digestibility of crossbred Angus bulls fed a high-concentrate diet. Twenty-one Angus beef bulls (497 ± 7.7 kg of initial BW, and 324 ± 3.0 d of age) were housed individually and fed ad libitum. Three experimental treatments were tested: complete feed of pellet and chopped straw in a single feeder (TMR), pellet and chopped straw fed separately in two feeders (SS); pellet and long unprocessed straw fed separately in two feeders (LS). Feed consumption, fecal and bloat scoring were recorded daily. Every 2 wk TMR sorting, eating behavior, general activity, social and oral behaviors, and BW were recorded. At days 14 and 35 apparent total tract digestibility during one week was measured. At days 28 and 42 rumen samples were collected via rumenocentesis to measure rumen pH and determine ruminal volatile fatty acids (VFA) concentrations. At the study end (day 57) animals at slaughterhouse liver and rumen wall color and lesions were recorded by a macroscopic inspection. No differences among treatments in total DMI, and fecal and bloat scoring were observed. The straw to concentrate ratio was smaller in the SS and LS treatments (8–92) than in the TMR (15–85), and sorting analyses indicated that TMR bulls refused large particles (> 4 mm) and small particles (
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