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A series of 28 compounds, 3-nitro-1H-1,2,4-triazole, were synthesized by click-chemistry with diverse substitution patterns using medicinal chemistry approaches, such as bioisosterism, Craig-plot, and the Topliss set with excellent yields. Overall, the analogs demonstrated relevant in vitro antitrypanosomatid activity. Analog 15g(R1= 4-OCF3–Ph, IC50= 0.09 μM, SI = >555.5) exhibited an outstanding antichagasic activity (Trypanosoma cruzi, Tulahuen LacZ strain) 68-fold more active than benznidazole (BZN, IC50= 6.15 μM, SI = >8.13) with relevant selectivity index, and suitable LipE = 5.31. 15gwas considered an appropriate substrate for the type I nitro reductases (TcNTR I), contributing to a likely potential mechanism of action for antichagasic activity. Finally, 15gshowed nonmutagenic potential against Salmonella typhimuriumstrains (TA98, TA100, and TA102). Therefore, 3-nitro-1H-1,2,4-triazole 15gis a promising antitrypanosomatid candidate for in vivo studies.

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Objective: Parathyroid scintigraphy is mandatory for the identification of hyperfunctioning parathyroid glands in hyperparathyroidism (HPT). The use of 99mTc-methoxy-isobutyl-isonitrile (MIBI) as radiopharmaceutical for parathyroid scintigraphy is considered the most valid and useful considering its uptake mechanism. Several MIBI-based radiopharmaceuticals are commercially available (i.e., MediMIBI, TechneMIBI, Stamicis). They seem to have similar physico-chemical characteristics and the choice between them is based on commercial criteria, even though some differences in qualitative scintigraphic results have been appreciated. Aims of the study were: first, to compare the scintigraphic quantitative data of MediMIBI, TechneMIBI, and Stamicis, particularly in the view of a personalized medicine approach; second, to investigate the potential effect of clinical-laboratory data on image quality using one of these radiopharmaceuticals. Methods: Patients with diagnosis of HPT, who underwent a parathyroid scintigraphy using one of the three MIBI-based radiopharmaceuticals between December 2018 and October 2020, have been retrospectively identified. Parameters derived from regions of interest (ROIs) drawn on three different sites were obtained: a reasonable parathyroid lesion detected, an area in the lateral neck considered as the background, and the hepatic dome as the site of MIBI physiological uptake. Laboratory and clinical data, such as serum calcium, PTH, vitamin D, and creatinine levels, as well as possible drug-mediated interferences were considered. Results: Among 250 patients included, 83 (33.2%) had the parathyroid scintigraphy using MediMIBI, 84 (33.6%) using TechneMIBI and 83 (33.2%) using Stamicis. The ROIs on the parathyroid uptake at early images, on the background, and on the hepatic dome were statistically different among the three groups (p< 0.05). No significant differences were found in the remaining semi-quantitative parameters among the three groups, not even considering clinical-laboratory data. Conclusions: Some differences in semi-quantitative parameters emerged among MIBI-based radiopharmaceuticals for parathyroid scintigraphy. This might justify the different qualitative scintigraphic results obtained using one or another of the cited radiopharmaceuticals.

This study evaluated the effects of maternal supplementation of Bacillus-based direct-fed microbials (DFM) on physiology and growth performance of Bos indicus-influenced cow-calf pairs. On d 0 (~139 d before expected calving date), 72 fall-calving, Brangus crossbred beef heifers (25 to 37.5% Bos indicus; 20 to 22 mo of age) pregnant with their first offspring were stratified by their initial body weight (BW; 431 ± 31 kg) and body condition score (BCS; 6.0 ± 0.36; scale 1 to 9), and randomly allocated into 1 of 12 bahiagrass pastures (1 ha and 6 heifers per pasture). Treatments were randomly assigned to pastures (6 pastures per treatment) and consisted of heifers supplemented with 1 kg/d of soybean hulls dry matter (DM) added (BAC) or not (CON) with a Bacillus-based DFM mixture (BovacillusTM; Chr. Hansen A/S, Hørsholm, Denmark) from d 0 to 242 (139 ± 4 d prepartum to 104 ± 4 d postpartum). Calves were weaned on d 242 (96 ± 30 d of age) and then allocated into 1 of 16 drylot pens and limit-fed the same concentrate at 3.25% of BW until d 319. Maternal treatment effects were not detected (P ≥ 0.29) for herbage allowance, forage nutritive value, and heifer activity on pasture. Heifer BCS on d 39 and 63 tended (P ≤ 0.09) to be greater, whereas heifer BCS on d 91 was greater (P = 0.01) for BAC vs. CON heifers and did not differ (P ≥ 0.20) between treatments on d 179 and 242. Plasma concentrations of glucose did not differ from d 0 to 63 (P ≥ 0.14) but were greater (P = 0.005) on d 179 and tended (P = 0.09) to be greater on d 242 for BAC vs. CON heifers. First offspring BW at birth, ADG from birth to early weaning, and BW weaning did not differ (P ≥ 0.19), but BW at drylot exit (d 319) was greater (P = 0.05) for BAC vs. CON calves. Maternal treatment effects were not detected (P ≥ 0.42) for calf serum concentrations of IgG at birth, and post-vaccination plasma concentrations of glucose, cortisol and haptoglobin. However, serum titers against bovine respiratory syncytial virus (BRSV) were greater (P = 0.04) for BAC vs. CON calves on d 287, whereas seroconversion against parainfluenza 3 virus (PI-3) was greater (P < 0.01) for BAC vs. CON calves on d 271. Thus, maternal supplementation of Bacillus-based DFM increased prepartum BCS gain and postpartum plasma concentrations of glucose of heifers and led to positive carryover effects on post-weaning BW gain and some indicators of humoral immune response to vaccination in their first offspring.

This experiment compared performance, health, and physiological responses of high-risk stocker cattle receiving a Bacillus subtilis derivative during a 90-d grazing period. Crossbred steers (n = 120) were obtained from an auction yard and transported to the experimental facility. Steer shrunk body weight (BW) was recorded at arrival (d -1). This value was averaged with BW recorded on d 0 to represent initial BW (236 ± 2 kg). On d 0, steers were ranked by BW and allocated to 1 of 12 pastures (4-ha pastures; 10 steers/pasture). Pastures were randomly assigned to receive daily supplementation with dried distillers’ grains at 1% of BW containing either: 1) Bacillus licheniformis and B. subtilis ingredient (BOV; 2 g/steer daily of Bovacillus, Chr Hansen A/S, Horsholm, Denmark) or 2) no additive (CON). Cattle received treatments from d 0 to 90 while housed in pastures containing stockpiled grass forage, as well as free-choice access to water and a commercial mineral + vitamin mix without ionophores. Steers were assessed for bovine respiratory disease (BRD) signs daily, and shrunk BW recorded on d 91. Blood samples were collected, and BW recorded on d 0, 14, 28, 56, and 90. No treatment differences were detected (P ≥ 0.19) for glucose, non-esterified fatty acids, β-hydroxybutyrate, haptoglobin, or insulin-like growth factor 1, whereas a treatment × day interaction indicated increased cortisol in BOV cattle on d 56 (P < 0.01). A treatment × day interaction was detected for serum concentrations of parainfluenza type 3 antibodies, which was greater in BOV steers on d 90 (P < 0.01). No treatment differences were detected (P ≥ 0.23) for serum concentrations of antibodies against other BRD pathogen. Average daily gain (ADG) tended to be increased (P = 0.09) in BOV compared with CON steers from d 56 to d 90 (0.858 and 0.760 kg/d, respectively, although did not impact overall ADG (P = 0.48). No treatment differences were detected for steer final BW (P = 0.43). Total BW change per pasture was greater in BOV compared with CON (790 vs. 550 kg BW change; P = 0.02). No treatment differences were detected (P = 1.00) for BRD incidence, although steers removed from the experiment for health reasons tended to be increased in CON compared with BOV (P = 0.08). Despite lacking differences in health or physiological responses, B. licheniformis and B. subtilis supplementation increased overall production of BW in high-risk stocker steers during a 90-d grazing period.

This experiment evaluated the effects of an analogue of bovine appeasing substance (BAS) on growth, stress, handling quality, and reproduction of Nellore heifers. Twenty-nine heifers [body condition score (BCS) = 4.84 ± 0.13; body weight (BW) = 371 ± 0.77 kg; 23 ± 2 months of age)] were assigned to 1 of 2 treatments: 1) Saline solution (0.9 NaCl; n = 14) or BAS (Secure Cattle, IRSEA Group, Quartier Salignan, France; n = 15). Both solutions were applied topically in the nuchal skin area (5 mL/heifers) at the beginning (d -9) of the fixed-time artificial insemination protocol (FTAI). Heifers were allocated in two different pastures according to the treatment (one pasture per treatment) for 15 d (time of action of BAS) and then combined in a single group (both treatments in the same pasture). Heifers were inseminated on d 0, and BW and BCS were collected on d -9 and 40. The temperament in the squeeze chute (entry and exit scores; scale 1 – 3; chute score; scale 1 – 5; greater values mean more reactive heifers) and blood samples were collected on d -9, 0, 7, 14 and 30. The percentual of aggression, accident, and dirtiness in the perineal region were evaluated on d 0 along with time to inseminate, estrus expression scores (scale 1 – 3; 1 = no expression, 2 = low expression, 3 = greater expression), and dominant follicle size. The corpus luteum size was measured on d 7 and 14, and the pregnancy and embryo size on d 40. The BAS vs. Saline-treated heifers tended to have greater BW change (23.6 vs. 20.8 ± 1.03 kg; P = 0.09) and had greater BCS change (0.05 vs. -0.21 ± 0.05; P < 0.01). The BAS vs. Saline-treated heifers tended to have lesser entry scores (1.42 vs. 1.67 ± 0.09; P = 0.09) and percentual of dirtiness in the perineal region (48.3 vs. 71.1 ± 8.67 %; P = 0.06), and no effects of treatment were detected (P ≥ 0.39) for chute score, exit score, aggression, accident and time to inseminate. The BAS vs. Saline-treated heifers had greater estrus scores (2.00 vs. 1.27 ± 0.21; P = 0.03), and no effects were detected (P ≥ 0.11) for ovarian variables, pregnancy rate, and conceptus size. Furthermore, no treatment effects were detected (P ≥ 0.25) for serum concentration of cortisol and plasma concentration of haptoglobin, ceruloplasmin, and progesterone. Therefore, BAS application at the beginning of the FTAI protocol could decrease the temperament and dirtiness in the perineal region and increase the growth and estrus expression of beef heifers.

This experiment compared performance, health, and physiological responses of weaned cattle receiving Bacillus subtilisderivative in a 90-d grazing period. Recently weaned Angus-influenced steers (n = 120) were obtained from an auction yard on d -1 and transported 150 km to research facility. Immediately after arrival on d -1, shrunk body weight (BW) was recorded and steers were grouped with access to native forage and water. On d 0, an additional body weight was recorded and combined with d -1 values to represent initial body weight (BW; 236 ± 2 kg). Steers were ranked according to initial BW and allocated to 1 of 12 pastures (4-ha pastures; 10 steers/pasture). Pastures were randomly assigned to receive daily supplementation with dried distillers grains at 1% of BW with either: 1) Bacillus subtilisingredient (BOV; 2 g/steer daily of Bovacillus; Chr Hansen A/S, Horsholm, Denmark), or 2) no additive (CON). Cattle received treatments for the duration of the experiment (d 0 to d 90) while housed in pastures containing stockpiled native forage, primarily Cynodon dactylonand Paspalum dilatatum, as well as free-choice access to water and a commercial mineral + vitamin mix not including ionophores. Bovine respiratory disease (BRD) signs were assessed daily throughout the experimental period. Steer unshrunk BW was recorded on d 14, 28, 56, and 90 and shrunk BW recorded on d 91 added 8% shrink as final BW. Blood samples and fecal swabs were collected from all steers on d 0, 14, 28, 56, and 90. Average daily gain (ADG) was increased in BOV groups compared with CON (0.858 and 0.760 kg/d, respectively) from d 56 to d 90 (P= 0.09) although this BW gain was not sufficient to impact final BW. Steer BW did not differ between treatments for all days (P≥ 0.43). Incidence of BRD was within the expected range for high-risk stocker cattle, although the number of sick animals was not reduced due to BOV supplementation (P= 1.00). Steers not completing the trial due to health-related complications tended to be increased in CON vs. BOV (P= 0.08). Differences in percentage of steers not completing the experiment, in conjunction with treatment effects for ADG in the final third of the experiment, increased the production total BW per BOV pasture by 43% (790 vs. 550 kg BW change; P= 0.02). Bacillus subtilissupplementation was effective in increasing production BWt during a 90-d grazing period despite no observed difference in BRD incidence.

Gastrointestinal pathogens cause productive, health, welfare, and financial losses to the beef cattle herd. Among these pathogens, Clostridium perfringenstype A is of interest given its prevalence, negative effects on intestinal health, and subsequent losses caused in beef animals. Based on this rationale, we hypothesized that a novel direct-fed microbial (DFM) formulation would support the gut barrier integrity upon a challenge with C. perfringens type A. Therefore, our objective was to evaluate how a novel DFM can support the in vitro integrity of intestinal epithelial cell monolayers exposed to C. perfringenstype A. Human cancer-derived epithelial intestinal Caco-2 cell monolayers were seeded in transwells for approximately 20 days. A DFM mixture containing Lactobacillus animalis, Propionibacterium freudenreichii, Bacillus licheniformis, and B. subtilis(BOVAMINE DEFEND Plus; Chr. Hansen A/S, Hørsholm, Denmark) was then administered to the apical side of the cell monolayers at 1.0 × 108CFU/transwell. Then, C. perfringenstype A (ATCC #13124) was added at 2.8 × 107CFU/transwell. Transepithelial electrical resistance (TEER) was measured over a 16-h period, after which area under the curve (AUC) was calculated. By itself, C. perfringenstype A led to a TEER decrease that reached its lowest levels at 6.5 h post-pathogen administration. Overall mean decrease in TEER was in the order of 19.3% when compared with control cells. The DFM mixture supported the integrity of the intestinal cell monolayers both with and without the C. perfringenstype A challenge, improving AUC by 24.5 and 19.1%, respectively, when compared with non-challenged control cells. In comparison with the negative control cells (C. perfringens-treated), improvements on AUC ranged from 45.6 to 39.4% vs. DFM and DFM + pathogen. In conclusion, these data support our hypothesis that a DFM mixture containing L. animalis, P. freudenreichii, B. licheniformis, and B. subtilis (BOVAMINE DEFEND Plus) supports the gut barrier integrity with or without the pathogenic challenge.

Ruminants are often exposed to pathogenic and non-pathogenic challenges during daily handling procedures and management activities. These responses, in turn, may trigger the release of pro-inflammatory compounds that can have negative effects on the host. Based on this rationale, we hypothesized that 1) pro-inflammatory cytokines (TNF-α and IFN-γ) would compromise the in vitro gut barrier integrity, and 2) a novel direct-fed microbial (DFM) formulation would support the gut barrier integrity under inflammatory conditions. Therefore, our objective was to evaluate the ability of a novel DFM mixture to support barrier integrity of intestinal epithelial cell monolayers in the absence or presence of TNF-α and IFN-γ by measuring transepithelial electrical resistance (TEER). Human cancer-derived epithelial intestinal Caco-2 cell monolayers were seeded on transwells for approximately 20 days. A DFM mixture containing Lactobacillus animalis, Propionibacterium freudenreichii, Bacillus licheniformis, and B. subtilis(BOVAMINE DEFEND Plus; Chr. Hansen A/S, Hørsholm, Denmark) at 1.0 × 108CFU/transwell was then administered to the apical side of the cell monolayers. Then, two hours later a cocktail containing TNF-α / IFN-γ at a 10:1 ratio (100:10 ng/mL, respectively) was added to the basolateral side after which TEER was measured over a 16-h period and used to calculate the area under the curve (AUC). Following the administration of the pro-inflammatory challenge, TEER was decreased, reaching its least concentrations at 5.5 h post-challenge, whereas overall AUC decreased by 21.4% compared with non-challenged control cells. The DFM mixture by itself was able to support the integrity of the intestinal cells by stimulating TEER increase over time and increasing overall AUC vs. non-treated cells (P< 0.01). Moreover, when cells receiving the DFM were challenged with the pro-inflammatory cocktail, TEER concentrations were still above 100% and no decrease was observed over the evaluation period (16 h). Moreover, when compared with non-DFM-treated but challenged cells, AUC was greater for DFM vs. challenged control cells (P< 0.05). In conclusion, these data support our hypothesis that a DFM mixture containing L. animalis, P. freudenreichii, B. licheniformis, and B. subtilis(BOVAMINE DEFEND Plus) supports the gut barrier integrity with or without the pro-inflammatory challenge.

The aim of this study was to investigate the effects of a probiotic containing B. licheniformisand B. subtilison fecal characteristics, blood parameters, and performance in feedlot cattle. One hundred and forty-four single-sourced Angus x Nellore bulls (initial shrunk BW = 401 ± 45.5 kg) were randomly assigned to 1 of 2 treatments (4 bulls per pen; 18 pen replicates per treatment). Treatments consisted of a finishing diet containing no probiotic (CONTROL) or a diet with 2 g of probiotic/bull/d (PROB). The probiotic was composed of B. licheniformisCH200 and B. subtilisCH201 (3.2 x 109CFU/g; BOVACILLUS). The diet was formulated using NASEM (2016) to provide an ADG of 1.9 kg/d (23% of corn silage, 65% of ground corn, 8% of soybean meal, 1% of urea, and 3% of mineral mix; DM basis). Bulls were adapted to the finishing diet over a 15-d period and fed for a total of 84 d. Fecal samples were collected on d 33 and 63. Blood samples were collected on the day of shipping to slaughter. Statistical analysis of experimental data was performed using the MIXED procedure of SAS 9.4. An α of 0.05 determined significance and an α of 0.06 to 0.10 was considered a tendency. No differences were obtained for initial BW, final BW, ADG, hot carcass, and dressing. Dry matter intake was reduced by 4% (P= 0.002) and G:F was increased by 6.3% (P= 0.036) in bulls consuming PROB diet. Regarding blood parameters, no differences were observed for glucose, haptoglobin, and LBP. There was a tendency of greater plasma urea (P= 0.09) in bulls consuming CONTROL diets. Fecal characteristics (DM, starch, N, pH, and scoring) were not different between treatments. Overall, dietary supplementation with B. licheniformis and B. subtilis improves feed efficiency in feedlot cattle.

Probiotics are purported to improve gastrointestinal tract health across a variety of species. Thus, we evaluated the effects of supplementing a blend of Bacillusspp. on performance and immune function in immature beef cattle. Crossbred heifers (n = 192) were stratified by initial body weight (196 ± 15.4 kg) and randomly assigned to 1 of 24 pastures (1.8 ha each) of Kentucky 31 tall fescue. Heifers were offered 2.84 kg daily of a commodity feed blend and the supplement contained either no probiotic (CON) or of Bacillus spp. (BAC). Bacillus spp. was offered at 3 grams per head per day. Heifers were weighed on consecutive days on d 0 and 1, along with d 84 and 85 to establish initial and final body weight. Interim weights were taken every 28 d. A humoral response to chicken egg albumin (OVA) was established following a subcutaneous injection with OVA. To assess heifers’ immune responses, plasma was taken from two heifers per pasture on d 1, 15, 29, 43, and 57. Plasma samples were measured for the presence of anti-OVA IgG, proinflammatory cytokines, anti-inflammatory cytokines, and chemokines. There were no trt*day interactions (P≥ 0.30) for plasma response variables. Concentrations of pro-inflammatory cytokines IL-1α and IFN-γ decreased (P≤ 0.01) in heifers supplemented with BAC. The anti-inflammatory cytokine IL-1 F5 decreased (P= 0.02) with BAC supplementation. Chemokines MIG and MIP-1β also decreased (P≤ 0.03) with BAC supplementation. Heifers fed BAC tended to display (P= 0.09) greater titers of anti-OVA IgG than the control. Body weight and average daily gain did not differ (P≥ 0.53) throughout the experiment. Under the conditions of this experiment, heifers supplemented with Bacillusspp. demonstrated modulated immune responses.

Direct-fed microbials have been used in an effort to improve livestock production efficiency and reduce environmental impact. Among other species, spore forming bacteria such as Bacillus spp. present handling and processing advantages to be used as a probiotic. Several studies have demonstrated positive effects of Bacillusspp. on dairy cattle production. However, there is lack of information about its effects on beef cattle. Thus, the objective of this study was to evaluate the effect of a probiotic composed by a mixture of B. subtilisand B. licheniformison beef cattle performance, nutrient utilization, and enteric methane emissions (CH4). For this purpose, a total of 108 Angus crossbred heifers (318 ± 40.4 kg of BW) were used in a generalized randomized block design. The heifers were fed a sorghum silage-based diet containing 1 of the 2 following treatments: 1) control (CTL, no additive), or 2) Bacillusspp. probiotic (BSL, 310 mg/kg of diet DM), in an attempt to provide an average of 3 g·animal-1·d-1. The experimental period consisted in 7 d of adaptation to the pens, followed by 77 d of performance data collection, 5 d of apparent total tract digestibility measurements, and 5 d of CH4measurements. No effect of treatment (P> 0.05) was observed for dry matter intake, average daily gain, or gain-to-feed ratio. The inclusion of the probiotic did not affect (P> 0.05) apparent total tract digestibility of dry matter, organic matter, neutral detergent fiber and crude protein, whereas starch digestibility decreased (P= 0.027) with the probiotic inclusion. No differences (P> 0.05) were observed for CH4between the two treatments. Accordingly, feeding Bacillusspp. did not affect cattle performance, nutrient utilization, or enteric CH4emissions in beef heifers when included at 310 mg/kg of dietary dry matter.

We evaluated the effects of supplementing direct-fed microbials (DFM), containing Bacillus licheniformisand Bacillus subtilis, on performance, rumen morphometrics, intestinal gene expression, and blood and fecal parameters in finishing bulls. Nellore × Angus bulls (n= 144; initial BW = 401 ± 45.5 kg) were distributed at random in 36 pens (4 bulls/pen and 18 pens/treatment), following a completely randomized design. A ground corn-based finishing diet was offered for ad libitum intake twice a day for 84 d, containing the following treatments: 1) control (without DFM); 2) DFM (B. licheniformisand B. subtilis) at 6.4 × 109CFU (2 g) per animal. The data were analyzed using the MIXED procedure of SAS, with a pen representing an experimental unit, the fixed effect of the treatment, and the random effect of pen nested within the treatment. For fecal parameters (two collections made), the collection effect and its interaction with the treatment were included in the model. Bulls that received the DFM had a decreased dry matter intake (P≤ 0.01), did not differ in average daily gain (2.05 kg; P= 0.39), and had a 6% improvement in gain:feed (P= 0.05). The other performance variables, final BW, hot carcass weight, and hot carcass yield, did not differ (P> 0.10). Plasma urea-N concentration decreased by 6.2% (P= 0.02) in the bulls that received DFM. Glucose, haptoglobin, and lipopolysaccharides were not different between treatments (P> 0.10). Ruminal morphometrics were not affected by the treatment (P> 0.10). The use of DFM tended to reduce fecal starch (P= 0.10). At slaughter, bulls fed DFM had an increased duodenal gene expression of tryptophan hydroxylase-1 (P= 0.02) and of superoxide dismutase-1 (P= 0.03). Overall, supplementation with DFM based on B. licheniformisand B. subtilisto Nellore × Angus bulls in the finishing phase decreased dry matter intake, did not influence ADG, improved gain:feed, and increased the expression of genes important for duodenal function.Supplementation with direct-fed microbials based on Bacillus licheniformisand Bacillus subtilisoffered to Nellore × Angus bulls in the finishing phase decreased dry matter intake, no influence on average daily gain, improved gain:feed, and an increase in expression of genes important for duodenal function.One of the main alternatives of additives to modulate the microbial population in the gastrointestinal tract (GIT), especially in the intestine, is the use of direct-fed microbials (DFM). This class of additives comprises all the feed products that contain a live or naturally occurring source of microorganism. The inclusion of DFM in diets of ruminants in the finishing phase may improve gain:feed by modifying the composition of the microbial community in the GIT to bring about a better symbiotic relationship with the host. These effects may be achieved with the use of Bacillusspp. bacteria, such as Bacillus licheniformisand Bacillus subtilis. Mixtures of these bacteria are able to foster positive effects in the finishing phase of beef cattle fed high-energy diets, which reinforces the need for studies that examine the effects and mechanisms of these species. In this study, feedlot Nellore × Angus bulls that received a DFM composed of B. licheniformisand B. subtilishad decreased dry matter intake, no influence on average daily gain, improved gain:feed, and an increase in expression of genes important for duodenal function.

This experiment evaluated the performance, health, and physiological responses of high-risk steers receiving a Bacillus-based probiotic during a 90-d grazing period. A total of 240 Angus-influenced steers were used in this experiment that was replicated over 2 yr (120 steers/year). Each year, steers were obtained from an auction yard and transported to the experimental facility (120 km). Steer body weight (BW) was recorded at arrival (day −1), and this value was averaged with BW recorded on day 0 to represent the initial BW (236.6 ± 1.5 kg). On day 0, steers were ranked by BW and allocated to 1 of 12 pastures with stockpiled native grass (4-ha pastures; 10 steers/pasture). Pastures were randomly assigned to receive daily supplementation with dried distillers’ grains at 1% of BW containing either: 1) Bacillus subtilis+ B. licheniformisprobiotic (BOV; 2 g/steer daily of Bovacillus; Novonesis, Horsholm, Denmark) or 2) no feed additive (CON). Cattle received treatments from days 0 to 90, in addition to free-choice access to water and mineral + vitamin mix without ionophore. Steers were assessed for bovine respiratory disease (BRD) signs daily. Blood samples were collected and full BW was recorded on days 0, 14, 28, 56, and 90. Shrunk BW was recorded on day 91 after 16 h of feed and water restriction, and a 4% pencil shrink was used to calculate the final BW. Average daily gain (ADG) was calculated based on initial and final BW. No treatment effects were detected (P ≥ 0.73) for steer final BW and ADG. A treatment × day interaction was detected (P ≤ 0.05) for plasma haptoglobin concentration, which was greater for CON steers on days 14 and 28 (P≤ 0.02). Incidence of BRD signs did not differ (P = 0.97) between treatments (51.7% and 51.3% for BOV and CON, respectively; SEM = 7.70). However, steer mortality + removals for health complications were greater (P= 0.01) in CON compared to BOV (0.00% vs. 5.04%, respectively; SEM = 1.41). Supplementing BOV improved (P ≤ 0.04) total pasture-based liveweight change (643 vs. 502 kg/pasture, respectively; SEM = 45) and final pasture-based total liveweight (3,007 vs. 2,869 kg/pasture, respectively; SEM = 46). Collectively, supplementation with a probiotic based on B. subtilisand B. licheniformisto high-risk stocker cattle did not alleviate the incidence of BRD signs nor improved ADG, but decreased acute-phase protein response, reduced steer mortality + removal, and increased pasture-based productivity during a 90-d grazing period.Stocker cattle are exposed to several stressors within a short period of time, which impair their immunity and lead to bovine respiratory disease. This study evaluated a Bacillus-based probiotic supplement to improve immunity and productivity of high-risk stocker cattle.Stocker cattle are exposed to several stressors within a short period of time, which impair their immunity and lead to bovine respiratory disease (BRD). With the increased regulations regarding the use of antimicrobials in cattle nutrition, novel dietary strategies to improve health and productivity of stocker cattle are warranted. One example is supplementing Bacillus-based probiotics, which promote performance and immunity in high-stress cattle. In this study, steers were purchased from a commercial auction yard soon after weaning, transported to the research facility, and assigned initial processing within a 48-h period. Steers were assigned to pastures and were supplemented or not with the Bacillus-based probiotic during a 90-d grazing period. In general, supplementing steers with the Bacillus-based probiotic did not impact growth rates or BRD incidence. However, no steers that received the Bacillus-based probiotic died from BRD consequences nor were removed from the experiment due to health reasons, whereas 5% of unsupplemented steers did not complete the 90-d experiment. Consequently, pasture-based liveweight gain was increased by 28% due to Bacillus-based probiotic supplementation. Results from this study indicate that supplementing a B. subtilis + B. licheniformis probiotic could be an alternative to improve the health and overall productivity of high-risk stocker cattle.

This study evaluated the effects of maternal supplementation of a Bacillus-based direct-fed microbial (DFM) on the physiology and growth performance of Bos indicus-influenced cow–calf pairs. On day 0 (~139 d before expected calving date), 72 fall-calving, Brangus crossbred beef heifers (20 to 22 mo of age) pregnant with first offspring were stratified by their initial body weight (BW; 431 ± 31 kg) and body condition score (BCS; 6.0 ± 0.36; scale 1 to 9), and randomly allocated into 1 of 12 bahiagrass pastures (1 ha and six heifers per pasture). Treatments were randomly assigned to pastures (six pastures per treatment) and consisted of heifers supplemented with 1 kg/d of soybean hulls (dry matter, DM) that was added (BAC) or not (CON) with DFM containing Bacillus subtilisand B. licheniformis(Bovacillus; Chr. Hansen A/S, Hørsholm, Denmark). Treatments were provided from days 0 to 242 (139 ± 4 d prepartum to 104 ± 4 d postpartum). Calves were weaned on day 242 (96 ± 30 d of age) and then allocated into 1 of 16 drylot pens and fed the same concentrate at 3.25% of BW (DM) until day 319. Maternal treatment effects were not detected (P≥ 0.29) for herbage allowance and forage chemical composition. Heifer BCS on days 39 and 63 tended (P ≤ 0.09) to be greater for BAC vs. CON heifers, whereas heifer BCS on day 91 was greater (P= 0.01) for BAC vs. CON heifers. Heifer BCS did not differ (P≥ 0.20) between treatments on days 179 and 242. Plasma glucose concentration did not differ from days 0 to 63 (P≥ 0.14) but were greater (P< 0.01) on day 179 and tended (P= 0.09) to be greater on day 242 for BAC vs. CON heifers. Calf BW at birth, ADG from birth to weaning, and BW at weaning did not differ (P≥ 0.19) between treatments, but calf BW at drylot exit (day 319) was greater (P= 0.05) for BAC vs. CON calves. Maternal treatment effects were not detected (P≥ 0.42) for calf serum concentration of IgG at birth and postvaccination plasma concentrations of glucose, cortisol, and haptoglobin. Serum titers against bovine respiratory syncytial virus (BRSV) were greater (P= 0.04) for BAC vs. CON calves on day 287, whereas seroconversion against parainfluenza-3 virus (PI-3) was greater (P< 0.01) for BAC vs. CON calves on day 271. Thus, maternal supplementation of a Bacillus-based DFM increased prepartum BCS gain and postpartum plasma glucose concentration of heifers and led to positive carryover effects on postweaning BW gain and humoral immune response in their offspring.Bacillus-based direct-fed microbials offered during late gestation and early lactation of Bos indicus-influenced beef heifers increased maternal body condition score at calving and led to positive effects on growth and humoral immune function in their offspring.Direct-fed microbials (DFM), such as Bacillusspp., have been shown to produce a wide variety of enzymes related to nutrient digestion and to support gastrointestinal tract immune function and integrity, leading to increased nutrient digestibility and cattle performance. Nutritional management of beef cows during gestation and early lactation has been associated with enhanced future offspring growth performance and immune response following birth. The present study combined the use of Bacillus-based DFM for pregnant heifers during critical production stages (late gestation and early lactation) to promote the overall performance of heifers and their offspring. Heifers offered Bacillus-based DFM had greater body condition score at calving and postpartum plasma concentration of glucose, whereas their offspring had similar body weight at birth, but greater growth performance when fed relatively high amounts of protein and energy in drylot compared to cohorts born from heifers that did not receive Bacillus-based DFM supplementation.

We conducted two experiments to evaluate the effects of a novel bacterial-based direct-fed microbial (DFM) on intestinal barrier integrity using the in vitro transepithelial electrical resistance (TEER) assay. In experiment 1, human-derived Caco-2 cells received or not (CON) a DFM containing Ligilactobacillus(formerly Lactobacillus) animalis506, Propionibacterium freudenreichii507, Bacillus paralicheniformis809, and B. subtilis597 (BDP; BOVAMINE DEFEND®Plus) at a rate of 1 × 108CFU/transwell. Concurrently with treatment application (CON or BDP), a pathogenic challenge of Clostridium perfringenstype A was added alone (PAT) or with BDP (PAT + BDP) at a rate of 2.8 × 107CFU/transwell in a 2 × 2 factorial arrangement. In experiment 2, Caco-2 cells were also assigned in a 2 × 2 factorial design to CON or BDP and then, 2 h post-treatment administration (CON and BDP), a mixture of tumor necrosis factor-alpha (TNF-α) and interferon-gamma (IFN-γ) was added alone (CYT) or with BDP (CYT + BDP) at a 10:1 ratio, respectively. In both experiments, TEER was measured for 18 h. In experiment 1, a DFM × pathogen × hour interaction was observed for TEER (P< 0.0001). Adding the PAT alone initially tended to increase TEER vs. CON from 1.1 to 2.2 h (P≤ 0.09), increased TEER at 3.2 h (P< 0.01), but reduced TEER from 5.4 to the end of the experimental period at 18.4 h (P≤ 0.01). On the other hand, adding DFM, with or without the pathogenic challenge, yielded greater TEER vs. CON-CON and CON-PAT for most of the experimental period (P≤ 0.04). A similar interaction was detected and reported in experiment 2 (P< 0.0001). The CYT challenge reduced mean TEER compared with all other treatments from 3.2 h to the remainder of the study (P≤ 0.03). On the other hand, BDP-CYT was able to maintain the integrity of the epithelial cells when compared with CON-CON throughout the experimental period (P≤ 0.03), the exception being at 3.2 h (P= 0.20). Moreover, BDP-CON increased (P≤ 0.04) TEER when compared with CON-CON from 3.2 to 18.4 h, but also in comparison with BDP-CYT from 4.3 to 18.4 h post-DFM and challenge administration into the cells. In summary, C. perfringenstype A and a pro-inflammatory cytokine cocktail compromised the integrity of intestinal epithelial cell monolayers in vitro, whereas adding a multispecies bacteria-based DFM counteracted these damaging effects.Both challenges, C. perfringenstype A and pro-inflammatory cytokine mixture, lead to a loss on the integrity of the intestinal epithelial cells. Adding a multispecies bacterial-based DFM to this in vitro setting supported the integrity of the cells following the challenges.Two experiments were designed to evaluate the effects of adding a bacterial-based direct-fed microbial (DFM) containing Lactobacillus animalis506, Propionibacterium freudenreichii507, Bacillus paralicheniformis809, and Bacillus subtilis597 on the integrity of intestinal epithelial cells challenged with Clostridium perfringenstype A or a pro-inflammatory cytokine cocktail. Regardless of the challenge, the addition of the DFM maintained the integrity of the intestinal epithelial cells in vitro. These results help to elucidate the potential beneficial effects that the bacterial-based DFM containing L. animalis506, P. freudenreichii507, B. paralicheniformis809, and B. subtilis597 may bring to livestock species.

Parathyroid scintigraphy is superior to other imaging techniques in detecting hyperfunctioning parathyroid glands. It is mainly performed using double-phase or dual-tracer subtraction methods. Neither of the techniques is perfect and different protocols are being used. We aimed to evaluate the accuracy of double-phase and subtraction methods in detecting abnormal gland as well as the potential effects of coexisting thyroid disease and clinical-laboratory data. We considered patients with primary hyperparathyroidism who underwent parathyroid surgery, after a parathyroid scintigraphy between April 2015 and February 2017. Sixty-eight patients were included; in 45 cases (66.2%), a thyroid disease was coexistent. Diagnostic performances of the two techniques were compared. The effect of thyroid disease and clinical-pathological data on examination interpretation was considered. Double-phase scintigraphy showed higher sensitivity and accuracy in detecting the exact abnormal gland compared to the digital subtraction (90% and 75% vs. 76% and 66%, respectively). For double-phase technique, sensitivity and accuracy were higher in cases with no thyroid disease when compared to those with thyroid disease (92% and 86% vs. 88% and 69%, respectively). Similarly, for digital subtraction, sensitivity and accuracy were higher in the absence of thyroid disease compared to their presence (84% and 79% vs. 70% and 58%, respectively). There was no significant variation in the performance of both techniques, considering clinical-laboratory data. Double-phase scintigraphy has been more accurate than digital subtraction. The presence of thyroid disease could be a possible limit, affecting the subtraction more than the double-phase technique. Clinical data did not influence the scintigraphic outcome.

Chalcones and flavonoids constitute a large family of plant secondary metabolites that have been explored as a potential source of novel pharmaceutical products. While the simulation of these compounds by molecular dynamics (MD) can be a valuable strategy to assess their conformational properties and so further develop their role in drug discovery, there are no set of force field parameters specifically designed and experimentally validated for their conformational description in condensed phase. So the current work developed a new parameter set for MD simulations of these compounds’ main scaffolds under GROMOS force field. We employed a protocol adjusting the atomic charges and torsional parameters to the respective quantum mechanical derived dipole moments and dihedrals rotational profiles, respectively. Experimental properties of organic liquids were used as references to the calculated values to validate the parameters. Additionally, metadynamics simulations were performed to evaluate the conformational space of complex chalcones and flavonoids, while NOE contacts during simulations were measured and compared to experimental data. Accordingly, the employed protocol allowed us to obtain force field parameters that reproduce well the target data and may be expected to contribute in more accurate computational studies on the biological/therapeutical role of such molecules.

Direct-fed microbials (DFM) support the health and performance of the herd through different modes of action, including improvements on nutrient digestibility. In fact, Bacillusspp. has been identified as an important enzyme-producing bacteria that could benefit overall nutrient utilization by the beef and dairy cattle herd. Based on this rationale, we hypothesized that incubation of a Bacillus-based DFM would improve nutrient digestibility of feedstuffs often fed to ruminants. Therefore, our objective was to evaluate the effects of a Bacillus-based DFM on in vitro nutrient digestibility of different single feedstuffs. Six individual feedstuffs were sampled from China and South Africa and used herein. Feedstuffs were incubated or not (CON) with a Bacillus-based DFM (DFM; BOVACILLUS; B. licheniformisand B. subtilis; 3.2 × 109CFU/gram; Chr. Hansen A/S., Hørsholm, Denmark) over a 48-h in vitro period. The dose of DFM was adjusted to the volume of the flasks being used and to represent the commercial dose of 3 grams/head per day (9.6 × 109CFU/head). In vitro gas production, dry matter (DM), and neutral detergent fiber (NDF) digestibility were evaluated at 24 and 48 h. Data were analyzed using PROC MIXED of SAS (version 9.4). Mean total in vitro gas production increased by 5.1 and 6.5% when DFM was incubated at 24 and 48 h, respectively (P< 0.01). At 24 h, NDF digestibility was 12.6% greater for DFM-treated feedstuffs vs. CON (P= 0.02; 27.7 vs. 24.6% for DFM and CON, respectively), but no effects were observed on DM digestibility (P= 0.26). Conversely, 48-h DM and NDF digestibility were greater for DFM vs. CON (P≤ 0.05), with improvements that were in the order of 3.7 and 15.1%, respectively. In conclusion, we observed greater in vitro fiber degradability and gas production of feedstuffs collected from China and South Africa over a 48-h period when the DFM BOVACILLUS was added at a dose representative of 9.6 × 109CFU per head. Greater improvements in fiber degradability are likely due to the enzymes produced by the DFM BOVACILLUS.

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This study evaluated productive and physiological responses in feedlot cattle receiving a finishing diet that included Ca salts of palm oil (CSPALM), or a blend of Ca salts of palm, cottonseed, and soybean oils (CSMIX). Ninety yearling steers were housed in 15 pens equipped with Calan-gate feeders (6 steers/pen). Steers within each pen were stratified by shrunk body weight (BW; 410 ± 3.3 kg across pens) on d 0 and assigned to receive a total-mixed ration (TMR) containing (dry matter basis) 2.2% of CSPALM (n= 30), 2.2% of CSMIX (n= 30), or no supplemental fat (CON; n= 30). Individual TMR intake was evaluated weekly. Blood samples were collected on d 0, 28, 56, 91, 119, and 147. Samples of the Longissimusmuscle (LM) were collected on d 84 via biopsy. Upon slaughter on d 148, hot carcass weight (HCW) was recorded to estimate final BW (63% dressing), and one LM steak sample (2.54 cm thickness) was removed from the right side of each carcass. Steer ADG was greater (P = 0.02) for CSMIX compared with CSPALM and tended to be greater (P = 0.09) for CSMIX compared with CON. The gain:feed ratio was greater (P ≤ 0.05) for CSMIX compared with CSPALM and CON, and carcass LM area was less (P = 0.01) for CSPALM compared with CSMIX and CON. No treatment effects were detected (P ≥ 0.21) for TMR intake, final BW, and other carcass merit traits including marbling. Mean plasma cholesterol concentrations were greater (P < 0.01) in CSMIX and CSPALM compared with CON, and mRNA expression of adipocyte fatty acid binding proteinin the LM on d 84 was greater (P≤ 0.04) in CSPALM compared with CSMIX and CON. No treatment effects were detected (P ≥ 0.15) for plasma concentrations of glucose, insulin, insulin-like growth factor I, and leptin, nor for other LM genes associated with marbling and muscle growth. Concentrations of total fatty acids (FA) in plasma and LM steak samples were greater (P < 0.01) in CSMIX compared with CSPALM and CON, and greater (P < 0.01) in the LM samples of CSPALM compared with CON. Steers receiving CSMIX had greater (P < 0.01) concentrations of polyunsaturated and ω-6 FA in plasma and LM steak samples compared with CSPALM and CON. Supplementing CSMIX improved gain efficiency and FA profile in the LM of feedlot steers compared with the CON diet, but the same responses were not observed when CSPALM was offered. Perhaps the advantages from CSMIX supplementation resulted from increasing the supply of polyunsaturated and ω-6 FA to the finishing diet.Supplementing saturated and monounsaturated fatty acids to cattle improve beef quality, whereas polyunsaturated fatty acids yield nutraceutical benefits to growth and health. Hence, this study evaluated a supplemental blend of saturated, monounsaturated, and polyunsaturated fatty acids to enhance performance and product quality of feedlot cattle.Supplemental fat has been provided to feedlot cattle to increase energy density of their diets, and may yield nutraceutical advantages if includes polyunsaturated fatty acids (FA). Alternatively, carcass quality can be improved when the fat supplement is based on saturated and monounsaturated FA. Hence, this experiment evaluated a blend of saturated, monounsaturated, and polyunsaturated FA to improve both performance and carcass merit in feedlot cattle. Steers received a finishing diet that included this blend (CSMIX), a source of saturated and monounsaturated FA (CSPALM), or no supplemental fat (CON). Growth rate and gain efficiency were improved in steers that received CSMIX compared with CSPALM and CON, and these traits did not differ between the latter treatments. Inclusion of CSMIX increased FA concentrations in the circulation of steers throughout the 147-day study and in Longissimusmuscle (LM) samples collected after slaughter. This increase in FA concentrations was associated with greater accumulation of polyunsaturated and ω-6 FA, suggesting that CSMIX resulted in LM with FA profile deemed more beneficial for human consumption. Collectively, supplementing CSMIX to feedlot steers improved gain efficiency and FA composition in the LM, and these advantages may be associated with increased supply of polyunsaturated ω-6 FA to the finishing diet.

Two experiments were designed to evaluate the effects of different probiotic combinations on rumen fermentation characteristics, performance, and carcass characteristics of feedlot Bos indicusbeef bulls offered a high-concentrate diet. In experiment 1, 30 rumen-fistulated Nellore steers were blocked by initial body weight (BW= 350 ± 35.0 kg) and within blocks (n= 10), animals were randomly assigned to receive: 1) high-concentrate diet without probiotic supplementation (n= 10; CONT), 2) CONT plus 1 g per head of a probiotic mixture containing three strains of Enterococcus faeciumand one strain of Saccharomyces cerevisiae(3.5 × 109CFU/g; n= 10; EFSC), and 3) CONT plus 2 g per head of a probiotic mixture containing Bacillus licheniformisand Bacillus subtilis(3.2 × 109CFU/g; n= 10; BLBS). The experimental period lasted 35 d, being 28 d of adaptation and 7 d of sampling. From day 34 to day 35 of the experimental period, ruminal fluid and fecal samples were collected every 3 h, starting immediately before feeding (0 h) for rumen fermentation characteristics and apparent nutrient digestibility analysis, respectively. In experiment 2, 240 Nellore bulls were ranked by initial shrunk BW (375 ± 35.1 kg), assigned to pens (n= 4 bulls per pen), and pens randomly assigned to receive the same treatments as in experiment 1 (n= 20 pens per treatment). Regardless of treatment, all bulls received the same step-up and finishing diets throughout the experimental period, which lasted 115 d. In both experiments, data were analyzed as orthogonal contrasts to partition-specific treatment effects: 1) probiotic effect: CONT vs. PROB and 2) probiotic type: EFSC vs. BLBS (SAS Software Inc.). In experiment 1, no contrast effects were observed on nutrient intake, overall nutrient digestibility, and rumen fermentation analyses (P≥ 0.13). Nonetheless, supplementation of probiotics, regardless of type (P= 0.59), reduced mean acetate:propionate ratio and rumen ammonia-N concentration vs. CONT (P≤ 0.05). In experiment 2, no significant effects were observed for final BW and dry matter intake (P≥ 0.12), but average daily gain and feed efficiency tended to improve (P≤ 0.10) when probiotics were offered to the animals. Probiotic supplementation or type of probiotic did not affect carcass traits (P≥ 0.22). In summary, supplementation of probiotics containing a mixture of E. faeciumand S. cerevisiaeor a mixture of B. licheniformisand B. subtilisreduced rumen acetate:propionate ratio and rumen ammonia-N levels and tended to improve the performance of feedlot cattle offered a high-concentrate diet.Rumen fermentation characteristics are positively modified when probiotics are fed to Bos indicusbeef cattle consuming a high-concentrate diet. The modulation of rumen fermentation might help to explain the positive and beneficial trends observed on performance of B. indicusfeedlot bulls offered different probiotics.Two experiments were designed to evaluate the effects of different probiotic combinations on rumen fermentation characteristics, performance, and carcass characteristics of feedlot Bos indicusbeef bulls offered a high-concentrate diet. The two probiotics consisted of a mixture containing three strains of Enterococcus faeciumand one strain of Saccharomyces cerevisiaeor a mixture of Bacillus licheniformisand Bacillus subtilis. Supplementation of probiotics, regardless of type, reduced acetate:propionate ratio, and mean rumen ammonia-N concentration and tended to improve the performance of feedlot cattle offered a high-concentrate diet, demonstrating the potential of this technology to be used as a feed additive for beef cattle.

The objective of this study was to evaluate the effects of a synthetic analogue of the bovine appeasing pheromone (i.e. bovine appeasing substance; BAS) on growth and temperament of heifers. At weaning (d 0), 30 heifers (Aberdeen Angus ′ Nelore; 8 ± 1 mo) were stratified by body weight (199.8 ± 16 kg) and randomly assigned to receive a single dose of BAS (n = 15; SecureCattle; Nutricorp, Araras, SP, Brazil) or saline (CON; n = 15; saline 0.9% NaCl). Treatments (5 ml) were topically applied to the nuchal skin area of each animal on d0. Body weight was collected on d 0, 6, 15, 45, and 150. Chute score (1 to 5; 1 = calm, no movement; 5 = violent and continuous struggling) and chute entrance and exit score (1 to 3; 1 = slow; 3 = fast) were collected on d 0, 2, 6, 15, 45 and 150. Scores were averaged across 4 trained technicians. Data were analyzed using the MIXED procedure of SAS. Heifers assigned to BAS had greater (P < 0.01) average daily gain (ADG) than heifers assigned to CON treatment from d 6 to 15 (2.35 and 1.88 kg/d ± 0.16) and from d 15 to 45 (1.79 vs. 1.56 kg/d ± 0.08). Heifers assigned to BAS had lower (P < 0.01) chute entrance score on d 6, 15, and 45, and chute exit score on d 2 to 45 (P = 0.05) when compared to heifers assigned to CON. Additionally, heifers assigned to BAS tended (P = 0.08) to have lower chute score, from d 1 to 150 than heifers assigned to CON. The application of BAS at weaning improved heifers ADG, likely due to an improvement in temperament, suggesting that BAS application has calming effects in the beef cattle herd.

This study, conducted at São Paulo University feedlot, Dracena, Brazil, was designed to evaluate the effect of adding either high-moisture corn, calcium salts of fatty acids (CSFA), or organic Zn+Cr on feedlot performance and carcass traits of Nellore cattle fed for 112 days. One-hundred fifty 18-mo-old Nellore bulls (404±24kg) were used and initially ranked, according to sires’ information, into groups of high and low EPD. Secondly, cattle were blocked by weight and randomly allocated to 30 pens (n = 5/pen), which were randomly assigned to the following 2x2 + 1 arrangement of treatments: Finely-ground corn, High-moisture corn, Finely-ground + CSFA, High-moisture corn + CSFA, and High-moisture corn + CSFA + Zn (90 ppm) and Cr (0.45 ppm). All diets contained 25 ppm of monensin. On day 0, one animal per pen was randomly selected to be slaughtered for baseline purposes. The replacement of finely-ground by high-moisture corn increased (P < 0.05) final BW (585 vs. 573 kg), ADG, HCW (321 kg vs. 315 kg) and decreased (P < 0.01) DMI (9.7 vs. 10.1 kg). Likewise, the addition of CSFA increased final BW, ADG (1.62 vs. 1.50 kg), and HCW (321 vs. 314 kg). The addition of Cr+Zn led to increased (P = 0.02) dressing percentage. No treatment effect was observed on final marbling (P > 0.55). It was observed interactions between treatments and EPD for G:F (P = 0.02) and final 12thrib-fat thickness (P = 0.03), where cattle from low-EPD groups needed high-moisture corn, CSFA and organic Zn+Cr to improve G:F and increase fat deposition; however, animals from high-EPD groups required only high-moisture corn. The adding of either high-moisture corn, CSFA, or organic Zn+Cr improved feedlot performance of Nellore cattle, and its effects seemed to be more effective in animals from low-EPD groups.

This experiment evaluated the effects of advancing days on feed (DOF) on insulin resistance (IR) of finishing Bos indicusbulls offered a high-concentrate diet. On day 0, 100 Bos indicusNellore bulls were housed in 12 feedlot pens (8 or 9 bulls/pen) for a 108-d feeding period, in a manner that all pens had an equivalent BW at the beginning of the finishing period. Bulls received the same diets throughout the 108-d feeding period. Diets were corn-based (38% starch) and were offered in amounts to ensure ad libitum intake. Individual shrunk BW was obtained on days 0 and 108 after 16 h of feed and water withdrawal, whereas unshrunk BW was recorded on days 19 and 60 for average daily gain (ADG) calculation. From days 0 to 108, feed intake and feed efficiency (FE) were calculated from each pen. Blood samples were collected on days 0, 60, and 108 to assess serum concentrations of haptoglobin, cortisol, glucose, and non-esterified fatty acids (NEFA), and insulin. Glucose, NEFA, and insulin concentrations were used to determine revised quantitative insulin sensitivity check index (RQUICKI), so that lower RQUICKI values indicate a greater IR of the herd. Pen was used as the experimental unit. Overall, DMI increased as DOF also increased (R2= 0.71), being greater from days 19 to 60 and 60 to 108 vs. 0 to 19 (P< 0.0001), but did not differ between days 19 to 60 and 60 to 108 (P= 0.79). Conversely, ADG and FE linearly decreased as DOF increased from days 0 to 108 (P< 0.0001; R2= 0.68 and 0.79, respectively). Log-transformed RQUICKI decreased as DOF increased up to 108 (P< 0.0001; r= −0.61). Similarly, serum concentrations of haptoglobin and cortisol increased as RQUICKI increased (P< 0.0001; r= 0.43 and 0.67, respectively). In summary, insulin resistance, per RQUICKI, increased and performance reduced in feedlot bulls with the advance of days on feed. Moreover, inflammatory markers were also positively associated with insulin resistance, suggesting that inflammation might be involved with the incidence of insulin resistance.This experiment was designed to evaluate the effects of feeding a high-starch during the feedlot phase (108 d) on performance, inflammatory markers, and insulin resistance of Bos indicusbulls. As feeding period increased, performance of the animals was reduced, whereas inflammatory markers were positively correlated with days on feed. Moreover, insulin resistance state also worsened as days on feeding increased, indicating that offering a high-starch diet for an extended period of time might lead to chronic inflammation and the occurrence of insulin resistance, which, in turn, could help us to explain the often observed decrease on performance of animals in the later stages of the feedlot phase.Insulin resistance was observed and worsened as days on feed increased in Bos indicusbulls receiving a high-starch diet.Performance and inflammatory markers were also associated with the occurrence of insulin resistance.

Sampling field soils to estimate soil water content and soil water storage (S) is difficult due to the spatial variability of these variables, which demands a large number of sampling points. Also, the methodology employed in most cases is invasive and destructive, so that sampling in the same positions at different times is impossible. However, neutron moderation, time domain reflectrometry, and, more recently, frequency domain reflectrometry methodologies allow measurements at the same points over long time intervals. This study evaluates a set of neutron probe data, collected at 15 positions placed randomly along a coffee crop contour line, over 2 years at 14-day intervals. The temporal stability of Swas again demonstrated, so that wetter or dryer locations remain so over time, and the definition of such positions in the field reduces the number of sampling points in future Sevaluations under similar conditions. An analysis was made to determine the minimum number of sampling points to obtain the average Sof the field within a chosen level of significance. Classical statistical analysis indicated that the 15 measurement positions could be reduced to four or even to one position to obtain a reliable field Saverage. State–time analysis showed Sestimations depend more on previous measurements of rainfall P(52%) than on evapotranspiration ET(28%) and S(20%). The analysis also showed that ETwas not realistically estimated from previous measurements of S; it was more dependent on previous measurements of ET(59%) than on P(30%) and S(9%). This statistical procedure showed great advantages over classical multiple regressions. Future studies of this type should be carried out at regularly spaced observation points in a grid, in order to allow a 2-D and 3-D state–space–time analysis.

Polymer electrolytes based on mixtures of poly(ethylene oxide-co-propylene oxide) and 1-methyl-3-propyl-imidazolium iodide (MPII) were investigated, aiming at their application in dye-sensitized solar cells (DSSC). The interactions between the copolymer and the ionic liquid were analyzed by infrared spectroscopy and 1H NMR. The results show interactions between the ether oxygen in the polymer and the hydrogen in the imidazolium cations. The ionic conductivities, electrochemical behaviors, and thermal properties of the electrolytes containing different concentrations of MPII were investigated. The electrolyte containing 70 wt % MPII presented the highest ionic conductivity (2.4 × 10−3S cm−1) and a diffusion coefficient of 1.9 × 10−7cm2s−1. The influence of LiI addition to the electrolytes containing different concentrations of MPII was also investigated. The DSSC assembled with the electrolyte containing 70 wt % MPII showed an efficiency of 3.84% at 100 mW cm−2. The stability of the devices for a period of 30 days was also evaluated using sealed cells. The devices assembled with the electrolyte containing less ionic liquid showed to be more stable.

We report a regioselective entry to 3-bromo- and 4-bromo-5-hydroxy-5H-furan-2-ones by photooxidation of 3-bromofuran with a singlet oxygen in the presence of a suitable base. By this procedure, a variety of 3-substituted -hydroxybutenolides have become for the first time easily accessible. Strategies employing these highly functionalized building blocks for the preparation of focused libraries of natural-like molecules are also discussed.

Metal−ligand (M−L) bond lengths for a range of ligands (carboxylates, chlorides, pyridines, water, tertiary phosphines, and alkenes) and a variety of metals have been retrieved from the Cambridge Structural Database, CSD. Analysis of the factors which affect M−L bond lengths (for example, ligand coordination mode, oxidation state, metal coordination number and geometry, spin and Jahn−Teller effects, and ligand trans to M−L bond) shows that it is generally possible to subdivide the M−L data sets systematically to obtain better defined, unimodal, bond length distributions with means and sample standard deviations (SSDs) which reflect the nature of the bond in question. Typically, the SSDs for the M−L data sets can be reduced to 0.04−0.05 Å by these methods. This work is an extension to tables of bond lengths in organometallic compounds and coordination complexes published in 1989. The importance of the factors which affect M−L bond lengths for particular metal−ligand groups are discussed. From the case studies reported, an algorithm is proposed by which compilation of a library of molecular geometry for metal complexes may be automated. The points that need to be considered to produce such a molecular library from the data stored in the CSD are discussed. The development of such a library would allow users to retrieve chemically well-defined geometric data rapidly and accurately. This should be of use, for example, to crystallographers and molecular modelers.

Three analogues of the natural bioactive cyclodepsipeptide jaspamide (3−5) were efficiently synthesized using a combination of solid and solution phase techniques. The preliminary design of the molecules has involved the rational substitution and/or simplification of the most critical structural features of the lead compound. The synthetic products were subjected to pharmacological assays, and the conformational properties were investigated by MM (molecular mechanics) and MD (molecular dynamics) calculations, to describe the potential pharmacophoric core responsible for the observed activities.

Chemical investigation of the Et2O extract of the marine sponge Rhaphisia lacazei resulted in the isolation of 13 pure bisindole alkaloids (1−13). Compounds (1−6) belong to the class of topsentins and have already been described. Compounds 7−13 are new products, closely related to the class of hamacanthins. The major compounds 1−3 were tested in vitro for antitumor activity; compounds 2 and 3 showed antiproliferative activity against human broncopulmonary cancer cells (NSCLC-N6) with an IC50 of 12 and 6.3 μg/mL, respectively.

The objective of this study was to evaluate the effects of a synthetic analogue of the bovine appeasing pheromone (i.e. bovine appeasing substance; BAS) on growth and temperament of weaned calves grazing Capim-Marandú (Urochloa brizantha cv Marandú). At weaning (d 0), 86 calves (47 steers and 39 heifers; Aberdeen Angus ′ Nelore; 8 ± 1 mo) were stratified by body weight (197.9 ± 24.9 kg) and randomly assigned to receive a single dose of BAS (n = 43; SecureCattle; Nutricorp, Araras, SP, Brazil) or saline (CON; n = 43; saline 0.9% NaCl). Treatments (5 ml) were topically applied to the nuchal skin area of each animal on d0. Body weight was collected on d 0, 8, 15, 51 and 100. Chute score (1 to 5; 1 = calm, no movement; 5 = violent and continuous struggling) and chute entrance and exit scores (1 to 3; 1 = slow; 3 = fast) were collected on d 3, 8, 15, 51 and 100. Scores were averaged across 3 trained technicians. Data were analyzed using the MIXED procedure of SAS. Calves assigned to BAS treatment had greater (P < 0.01) average daily gain (ADG) from d 8 to 15 (0.158 and -0.284 kg/d ± 0.07). Calves assigned to BAS treatment tended (P = 0.10) to have lower chute entrance score on d 8 and 51 and had lower (P = 0.02) chute exit score on d 8 than calves assigned to CON treatment. Additionally, calves assigned to BAS treatment tended (P = 0.07) to have lower chute score on d 8 when compared to calves assigned to CON treatment. The application of BAS at weaning improved calf ADG and improved calf temperament as observed by chute score and chute entrance and exit scores, suggesting that BAS application has calming effects.

This study was designed to evaluate the timing of administration of the bovine appeasing substance (BAS) on performance and physiological responses of Bos indicusfeedlot cattle. Nellore bulls (n= 100) were ranked by initial body weight (BW; 341 ± 18.5 kg) and assigned to receive BAS (n= 50) or placebo (CON; n= 50) on day −2 of the experiment. Treatments (5 mL) were applied topically to the nuchal skin area of each bull. Bulls were loaded into commercial livestock trailers immediately after treatment administration, transported for 880 km, and unloaded on day −1 at a commercial feedyard. On day 0, bulls within each treatment were again assigned to receive, in a 2 × 2 factorial arrangement, BAS or CON as described previously (25 bulls/treatment combination). Upon treatment administration on day 0, bulls were housed in 12 feedlot pens (3 pens/treatment) for a 108-d feeding period, which was divided into an adaptation (days 0–19), growing (days 20–60), and finishing (days 61–108) phases. Dry matter intake (DMI) was measured daily from days 0 to 108, whereas blood samples and hair from the tail switch were collected on days −2, 0, 19, 60, and 108. Administration of BAS prior to loading (day −2) improved ADG, FE, and DMI during adaptation and across the 108-d feeding period (P≤ 0.08), resulting in greater (P= 0.03) hot carcass weight and dressing percentage upon slaughter on day 109. A treatment × day interaction was detected for serum glucose concentrations (P= 0.05), which was greater (P= 0.03) on day 60 of the feeding period in bulls receiving CON prior to loading. Administration of BAS at feedlot entry (day 0) improved DMI, ADG, and FE during adaptation (P≤ 0.05), but it did not impact (P≥ 0.18) performance and carcass traits during the 108-d feeding period. Bulls administered BAS prior to loading and at feedlot entry had less (P≤ 0.05) mean serum cortisol concentrations across the 108-d feeding period (loading × feedlot entry interaction; P= 0.10) and greater (P≤ 0.05) serum insulin concentration on day 60 (loading × feedlot entry × day interaction; P= 0.05). In summary, BAS administration prior to loading increased the overall feedlot performance of Nellore bulls. These outcomes were noted in bulls that received or not a second BAS administration at feedlot entry, suggesting that the benefits of BAS are exploited when this substance is administered before transport to the feedlot.

We evaluated the influence of amount and crude protein (CP) supplementation frequency (SF) on nitrogen (N) use by wethers and the performance of late-gestation beef cows. In exp. 1, seven Western whiteface wethers (31.8 ± 1.4 kg) were used in an incomplete 7 × 4 Latin square to evaluate intake and N use. Wethers received one of the seven treatments in a 2 × 3 factorial design containing two levels of supplemental soybean meal offered at a rate of 100% (F) or 50% (H; 50% of F) of the estimated CP requirement daily, once every 5, or once every 10 d, plus a non-supplemented control (CON). Low-quality cool-season forage (4.9 % CP; dry matter [DM] basis) was provided daily for ad libitum intake. Experimental periods lasted 30 d. In exp. 2, 84 Angus × Hereford cows (560 ± 35 kg) were stratified by age, body condition score (BCS), and expected calving date and allocated to 1 of the 21 feedlot pens (three pens per treatment). Pens were randomly assigned to receive the same treatments as in exp. 1 and cows had free access to low-quality cool-season forage (2.9% CP; DM basis). Cow body weight (BW) and BCS were measured every 14 d until calving and within 24 h after calving. In exp. 1, supplementation did not alter total DM and organic matter (OM) intake (P≥ 0.26), but both parameters linearly decreased as SF decreased (P= 0.02). Supplementation increased DM, OM, and neutral detergent fiber (NDF) digestibility (P≤ 0.02). Additionally, F feeding linearly increased DM, OM, and NDF digestibility as SF decreased (P≤ 0.04). Digestibility of N, N balance, and digested N retained were greater with supplementation (P< 0.01), and N digestibility linearly increased as SF decreased (P= 0.01). Mean plasma urea-N concentration was not only greater (P< 0.01) for supplemented vs. CON wethers but also greater (P= 0.03) for F vs. H. In exp. 2, pre-calving BCS change was greater (P= 0.03) for supplemented cows. A linear effect of SF × supplementation rate for pre-calving BCS change was noted (P= 0.05), as F-supplemented cows lost more BCS compared with H as SF decreased. When considering supplementation intervals greater than 5 d, reducing the quantity of supplement provided, compared with daily supplementation, may be a feasible management strategy to maintain acceptable nutrient use and animal performance while reducing supplement and labor costs.

This experiment evaluated the influence of protein supplementation frequency (SF) and amount offered on intake, nutrient digestibility, and ruminal fermentation by rumen-fistulated beef steers consuming low-quality [2.9% crude protein (CP); dry matter (DM) basis], cool-season forage. Seven Angus × Hereford steers (300 ± 27 kg) fitted with ruminal cannulas were randomly assigned to 1 of 7 treatments in an incomplete 7 × 4 Latin square. Treatments, in a 2 × 3 factorial design plus a non-supplemented control (CON), consisted of 2 levels of supplemental soybean meal, 100% (F) or 50% (H) of the estimated rumen-degradable protein requirement, provided daily (D), once every 5 d (5D), or once every 10 d (10D). Experimental periods were 30 d and dry matter intake (DMI) was measured from days 19 to 28. On days 21 (all supplements provided) and 30 (only daily supplements provided; day immediately prior to supplementation for 5D and 10D treatments) ruminal fluid was collected for ruminal pH, ammonia-N (NH3), volatile fatty acids (VFA), and determination of ruminal fermentation variables. Forage and total DM, organic matter (OM), and nitrogen (N) intake increased with supplementation (P≤ 0.04). However, a linear effect of SF × amount of supplement interaction was observed for forage and total DM, OM, and N intake (P≤ 0.04), with each variable decreasing as SF decreased, but the decrease being greater with F vs. H. Apparent total tract DM, OM, and neutral detergent fiber digestibility was not affected by supplementation or amount of supplement provided (P≥ 0.10). In contrast, N digestibility increased with supplementation and for F vs. H (P< 0.01). Digestibility of DM, OM, and N increased linearly as SF decreased (P≤ 0.03). When all supplements were provided, ruminal NH3, total VFA, and molar proportions of all individual VFA increased with supplementation (P≤ 0.04), whereas acetate:propionate ratio decreased (P< 0.01). When only daily supplements were provided, none of the aforementioned fermentation parameters were affected (P≥ 0.09). In summary, reducing the amount of supplemental CP provided to ruminants consuming low-quality forages, when supplementation intervals are >5 d, can be a management tool to maintain acceptable levels of DMI, nutrient digestibility, and ruminal fermentation while reducing supplementation cost.

A new pyrroloiminoquinone alkaloid (1) belonging to the makaluvamine family has been isolated from the sponge Zyzzya cf. fuliginosa collected in the waters off the Vanuatu Islands. The compound, designated makulavamine P, was characterized on the basis of its spectral data and displayed cytoxicity in the μM range on KB cells and antioxidant activity.

The occurrence of 8,11,12-trihydroxyeicosa-5,14,17(Z),9(E)-tetraenoic acid and 8,11,12-trihydroxyeicosa-5,14(Z),9E-trienoic acid in starfish species, i.e.Patiria miniata, Dermasterias imbricata, Pycnopodia helianthoides, Culcita novaeguineaandNardoa tubercolatais reported.

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