Protein Concentration Research Articles

Physiological effects of interacting native and invasive bivalves under thermal stress

AbstractAcross many ecosystems in North America and Europe, native freshwater bivalves (Order Unionida) are threatened by fouling and competition for food by the invasive zebra mussel Dreissena polymorpha. In light of climate change, knowledge on the influence of water temperature on these competitive effects is important, yet poorly understood. This study examines the physiological impact of the interaction between D. polymorpha and the native European unionid Anodonta cygnea over a 28 day—period in response to water temperatures of 12, 19, and 25 °C by comparing their glycogen, glucose, lipid and protein concentrations. The laboratory experiment comprised three treatments: (1) fouling of A. cygnea by D. polymorpha, (2) both species present but not fouling; and (3) a control in which A. cygnea and D. polymorpha were placed separately. Increased water temperatures caused physiological stress in D. polymorpha as evident from reduced glycogen, glucose, lipid and protein concentrations. Dreissena polymorpha benefited from fouling of unionids, as individuals that fouled A. cygnea tended to have increased glycogen, glucose, lipid and protein concentrations. Competitive effects of D. polymorpha over the unionid bivalve species, however, were not intensified by elevated temperatures. Glochidia release, lower infestation intensity, and physiological stress of Dreissena at higher temperatures were likely confounding factors. The results of this study suggest that understanding the physiological consequences of species interactions at changing temperatures can be an important tool to assess future climate change impacts on freshwater bivalves and aquatic community structures.

168 Effects of feeding barley-based diets differing in fermentable starch and fiber on nutrient digestion and body composition in weaned pigs

Abstract Hulled and hull-less barley grains containing low insoluble fiber are alternative energy sources in weaning diets but are typically not part of weaned pig diets. The range in fermentable starch and fiber content among barley cultivars presents opportunities to investigate whether these fermentable carbohydrates of barley grain do influence nutrient digestion, gut short chain fatty acid (SCFA) profile and body composition. To explore, pigs [n = 40; body weight (BW) = 8.6 kg; 1-wk post-weaning] were housed individually and fed 1 of 4 diets containing 600 to 705 g of cereal grain/kg: 1) low-fermentable wheat (LFW); 2) high β-glucan, hull-less barley (HFBB); 3) high amylose, hull-less barley (HFAB); or 4) low-fermentable hulled barley (LFB). Diets provided 10.2 and 9.8 MJ NE/kg, and 1.32 and 1.21 g standardized ileal digestible (SID) lysine/MJ NE for phase 1 (d 0–14) and phase 2 (d 15–28), respectively. At the end of the trial, feces were collected, and pigs were slaughtered to collect digesta and the carcass that was subsequently ground. Subsamples of feces, digesta, and carcasses were freeze-dried and analyzed. The apparent ileal digestibility (AID) of crude protein (CP) and gross energy (GE) did not differ among diets. The AID of β-glucan was 3 times greater (P < 0.05) for LFW than HFBB and HFAB, while AID of starch was 12% less (P < 0.05) for HFAB than the other diets. Apparent total tract digestibility (ATTD) of GE and CP was 4.2 and 8.9% less (P < 0.05), respectively, for the 3 barley diets than the wheat diet. The apparent hindgut fermentation (AHF) of β-glucan was 2.6 times greater (P < 0.05) for HFAB and HFBB than LFW, while AHF of starch was 7.5 times greater for HFAB than LFW. The calculated NE value was greatest (P < 0.05) for LFW, intermediate for HFBB and HFAB, and least for LFB. Among diets, HFAB increased (P < 0.05) the concentration of butyrate (indicator of gut health) and propionate in cecal and colonic digesta, with butyrate up to 50% more (P < 0.05) than that of LFW. Carcass protein and lipid concentration did not differ among pigs fed the 4 diets. In conclusion, feeding barley instead of wheat grain to weaned pigs reduced diet nutrient digestibility but did not affect carcass protein and lipid concentration. Feeding high amylose barley increased gut SCFA concentration. Barley grain with increasing fermentable carbohydrates can thus be included as alternative ingredient in diets of young pigs.

157 Effects of increasing dietary protein concentration on digestibility of amino acids by growing pigs

Abstract Two experiments were conducted to test the hypothesis that the standardized ileal digestibility (SID) of amino acids (AA) by growing pigs is not influenced by dietary protein concentration if all AA are supplied by soybean protein isolate (SPI) or casein. In Exp. 1, cannulated pigs [n = 12; average body weight (BW) = 39.22 kg; SD = 3.96] were allotted to a replicated 6 × 3 Youden square design with the 6 diets and 3 periods. Five diets were formulated to contain 7 to 35% SPI. An N-free diet was also used. In Exp. 2, cannulated pigs (n = 24 average BW = 23.12 kg; SD = 2.89) were allotted a quadruplicated 6 × 2 Youden square design with the 6 diets and 2 periods. Five diets were formulated to contain 6.75 to 33.75% casein. An N-free diet was also used. Ileal digesta were collected and analyzed for dry matter (DM), Cr, and AA. The statistical models included diet as a fixed variable and square, animal, and period as random effects. Polynomial contrasts were used to test linear and quadratic effects of increasing dietary protein. Results from Exp. 1 indicated that the apparent ileal digestibility (AID) of DM linearly (P = 0.001) decreased and the AID of all AA quadratically (P < 0.05) increased as the inclusion of SPI increased in the diets (Table 1). The SID of His and Trp was not affected by the inclusion of SPI in the diets. However, the SID of Arg, Leu, Lys, Phe, Thr, and Val linearly (P < 0.05) decreased, and the SID of Ile and Met tended to linearly (P < 0.10) decrease as the inclusion of SPI in the diets increased. Results from Exp. 2 indicated that the AID of DM tended to linearly decrease (P = 0.089) as the inclusion of casein increased in the diets (Table 2). The AID of most AA increased quadratically (P < 0.01) and the AID of Trp tended to increase (quadratic, P = 0.068) with increasing inclusion rate of casein. The SID of most AA was independent of casein levels. In conclusion, the anti-nutritional factors may have been increased in the diets as the inclusion of SPI increased and thus resulted in reduced SID values. However, the SID values were not influenced by increasing dietary protein if all AA were supplied by casein that does not contain anti-nutritional factors. Therefore, it is concluded that pigs have the capacity to absorb peptides and AA with the same efficiency if the protein concentration is 30% as if it is 6%. Dietary protein concentration itself, therefore, is not likely to limit AA absorption. However, even small amounts of anti-nutritional factors appear to negatively impact the SID of AA.

155 Standardized ileal digestibility of amino acids in soybean meal, fermented soybean meal, and soy protein concentrate fed to nursery pigs

Abstract Accurate information of amino acid (AA) digestibility in protein supplements is critical in formulating diets for nursery pigs. Soybean products are widely used in nursery pig diets due to the high crude protein (CP) concentrations. The objective of the present study was to determine the standardized ileal digestibility (SID) of AA in soybean meal, 3 sources of fermented SBM (FSBM), and soy protein concentrate (SPC) fed to nursery pigs. Ten barrows with an initial body weight of 10.5 kg (standard deviation = 0.9) were surgically fitted with T-cannulas at the distal ileum. The animals were assigned to a replicated 5 × 5 Latin square design with 10 animals, 5 experimental diets, and 5 periods to obtain 10 observations per diet. Five diets were prepared to contain SBM (48.5% CP), 3 sources of FSBM (A, B, and C; 52.6% CP, 55.1% CP, and 49.8% CP, respectively), or SPC (58.4% CP) as the sole source of AA. Each period consisted of 4 d of adaptation period and 3 d of ileal digesta collection period. The values of basal endogenous losses of AA reported in the literature were used to calculate the SID of AA by correcting the apparent ileal digestibility (AID) values. Additionally, a 2-step in vitro procedure modified for nursery pigs was conducted to determine the in vitro ileal disappearance of CP in the 5 ingredients. The AID of all indispensable AA except His and Lys in FSBM-A and FSBM-B was greater (P < 0.05) than that in SBM, whereas AID values for all AA except Trp in SPC did not differ from the values in SBM. The SID of Leu and Trp in FSBM-A and FSBM-B was greater (P < 0.05) than that in SBM or SPC. The SID of all AA in SPC was not different from that in SBM. In vitro ileal disappearance of CP in 3 sources of FSBM was greater (P < 0.001) than that in SBM or SPC. In conclusion, in vitro ileal disappearance of protein was greater in fermented soybean meal compared with soybean meal or soy protein concentrate whereas the standardized ileal digestibility of most amino acids was comparable among the soybean-derived ingredients.

PSI-13 Digestible and metabolizable energy concentrations in soybean meal, fermented soybean meal, and soy protein concentrate fed to nursery pigs

Abstract Accurate information of energy concentrations in protein supplements is critical in formulating diets for nursery pigs. Soybean products are widely used in nursery pig diets due to the high crude protein (CP) concentrations. The objective of the present study was to determine the digestible (DE) and metabolizable energy (ME) in soybean meal (SBM, 48.5% CP), 3 sources of fermented SBM (FSBM-A, FSBM-B, and FSBM-C; 52.6% CP, 55.1% CP, and 49.8% CP, respectively), and soy protein concentrate (SPC, 58.4% CP) fed to nursery pigs. Twelve barrows with an initial body weight of 8.7 kg (standard deviation = 0.5) were individually housed in metabolism crates. A replicated 6 × 4 incomplete Latin square design with 12 animals, 6 experimental diets, and 4 periods was employed. A basal diet contained 96.9% corn as the sole source of energy. The experimental diets were prepared by replacing the basal diet with SBM at 31%, FSBM-A at 27%, FSBM-B at 26%, FSBM-C at 29%, and SPC at 24%. Each period consisted of a 4-d adaptation period and a 4-d collection period. The marker-to-marker procedure was used for the quantitative collection of feces. Urine was collected for 96 h. The difference procedure was employed to calculate the DE and ME values of the soybean ingredients. Additionally, a 3-step in vitro assay was conducted to determine in vitro total tract disappearance of nutrients in the ingredients. The DE concentrations in FSBM-A (4,171 kcal/kg as-fed), FSBM-B (4,111 kcal/kg as-fed), FSBM-C (4,027 kcal/kg as-fed) and SPC (3,999 kcal/kg as-fed) were greater (P < 0.05) than that in SBM (3,724 kcal/kg as-fed). The ME concentrations in FSBM-A (3,883 kcal/kg as-fed) and FSBM-B (3,841 kcal/kg as-fed) were greater (P < 0.05) than that in SBM (3,528 kcal/kg as-fed) but did not differ from those in FSBM-C (3,728 kcal/kg as-fed) or SPC (3,767 kcal/kg as-fed). The DE:gross energy (GE) was greatest (P < 0.05) in FSBM-B but lowest (P < 0.05) in SBM with intermediate ratios for other 3 ingredients whereas the ME:GE and ME:DE were not different among the 5 ingredients. The in vitro total tract disappearance of dry matter and organic matter in SPC was less (P < 0.05) than that in other ingredients. In conclusion, fermented soybean meal and soy protein concentrate contain greater energy concentrations compared with soybean meal.

73 Effects of blend of organic acids and monoglycerides on immune responses of weaned pigs experimentally infected with enterotoxigenic Escherichia coli F18

Abstract Our previous research implies that dietary supplementation of organic acids blend, monoglycerides blend, and the combination of both can mitigate diarrhea of weaned pigs infected with Escherichia coli (E. coli) F18. The objective of this study was to investigate the effects of dietary supplementation with a blend of organic acids, a blend of monoglycerides, or the combination of both on immune responses of weaned pigs experimentally infected with E. coli F18. Forty pigs [average body weight (BW) = 7.81 ± 0.84 kg] were individually housed and randomly assigned to one of four treatment groups with 10 replicate pigs per treatment. The four dietary treatments were: () Control: nursery basal diet, 2) Acids: basal diet supplemented with a blend of organic acids (formic acid, lactic acid, and sodium formate) at 0.3%, 3) Monoglycerides: basal diet supplemented with a blend of monoglycerides of fatty acids at 0.3%, or 4) Combination of acids and monoglycerides: basal diet added with the acids blend at 0.2% and the monoglycerides blend at 0.2%. The experiment lasted 28 d, with 7 d before and 21 d after the first inoculation (d 0). All pigs were orally inoculated with E. coli F18 (1010 CFU/3 mL) for 3 consecutive days from d 0. Serum samples were collected on d 0, 2, 5, and 14 post-inoculation (PI) to measure the concentration of C-reactive protein (CRP) and haptoglobin using ELISA. Porcine Cytokine/Chemokine13-Plex Discovery Assay (Eve Technology, Canada) was also conducted with serum samples from d 0, 5, and 14 PI. Data were analyzed by ANOVA using PROC MIXED of SAS with a randomized complete block design. The concentrations of serum CRP and haptoglobin were increased (P < 0.0001) on d 2 PI and then gradually decreased on d 5 and 14 PI. However, no differences were observed in serum CRP and haptoglobin concentrations among all treatment groups throughout the experiment. Pigs fed with monoglycerides tended (P < 0.10) to have decreased serum granulocyte-macrophage colony-stimulating factor (GM-CSF), interleukin 1-alpha (IL-1α), and tumor necrosis factor-alpha (TNF-α) concentrations on d 0 PI compared with other treatments. Pigs supplemented with monoglycerides also tended (P < 0.10) to have decreased GM-CSF on d 5 PI and less IL-1α on d 14 PI among all treatments. Supplementation of the combination tended (P < 0.10) to have greater GM-CSF concentration on d 5 PI compared with other treatments. In conclusion, dietary supplementation of monoglycerides blend or the combination of organic acids and monoglycerides alleviates systemic inflammation of weaned pigs during E. coli F18 infection by modulating related cytokines. However, additional research is needed to understand the modulation mechanisms.

PSIV-A-11 Potential of black soldier fly (Hermetia illucens) larvae frass as a functional ingredient for nursery pigs

Abstract Frass is the leftover material from the production of black soldier fly larvae (BSFL), which includes chitin-containing exoskeleton shedding, small, processed larvae, leftover dietary substrate, and larvae excrement. A total of 120 nursery pigs (initial body weight of 6.85 ± 0.10 kg) weaned at 21 d of age were used in a 6-wk study to evaluate the effect of increasing dietary levels of frass on growth performance, diarrhea score, and serological indices. Pigs were randomly assigned within sex and body weight blocks to 1 of 3 dietary treatments using 30 pens (4 pigs/pen; 10 replicates/treatment). Diets were prepared from a common basal and fed in 3 dietary phases of 2 wk each and included: 1) a control diet based primarily on corn and soybean meal; 2) the control with 3% added frass; and 3) the control with 6% added frass. On d 4, 7, and 14, individual fecal samples were obtained using fecal loops and were evaluated on a scale of 1 to 5 representing hard, normal, soft, watery soft, and watery liquid feces, respectively. Blood samples were collected on d 42 from 1 median pig per pen for the analysis of serum chemistry. Data were analyzed using the mixed model procedures as a randomized complete block design using linear and quadratic contrast comparisons to determine the effects of frass. Supplementation of frass linearly increased average daily gain (ADG) from d 7 to 14 (P = 0.044; 347.9, 378.8, and 396.8 g/d for 0, 3, and 6% frass, respectively), and tended to linearly increase ADG during Phase 1 (P = 0.079; 265.1, 284.8, and 298.6 g/d). No differences were observed for final body weight (P ≥ 0.146; 29.90, 31.48, and 30.70 kg for 0, 3, and 6% frass, respectively), and overall ADG, average daily feed intake (ADFI), or gain to feed ratio (G:F). Diarrhea scores increased (P < 0.001) from d 4 (2.96) to d 7 (4.20) and then decreased (P < 0.001) on d 14 (3.69). Inclusion of frass at 6% had a decreased diarrhea score compared with 3% frass on d 7 (P = 0.029), but pigs fed 6% frass had a greater diarrhea score (P = 0.038) than pigs fed the control diet on d 14. However, scores were indicative of diarrhea for all treatments on d 7 and 14. Serum concentrations of total protein, albumin, globulin, aspartate aminotransferase, alanine aminotransferase, alkaline phosphatase, γ-glutamyltranspeptidase, urea N, creatinine, glucose, Ca, P, Mg, K, Na, Cl, triglycerides, amylase, cholesterol, and creatine phosphokinase measured on d 42 were not different (P > 0.093) between treatments. All serological analytical results were generally within normal expected ranges. In conclusion, these findings suggest that dietary inclusion of frass may improve growth performance of nursery pigs, especially during the early stages of the nursery period.

144 Bioavailability of calcium salt of hydroxy analogue of methionine relative to DL-methionine to support growth performance of 11 to 20 kg pigs

Abstract Methionine (Met) is typically the third limiting AA in commercial swine diets and is commonly supplied as dry DL-Met (99% pure) or DL-Met hydroxy analogue. Data on the relative bioavailability (RBV) of calcium salt of hydroxy analogue of DL-Met (MHA-Ca; 84%) compared with DL-Met in weaned pigs is scarce. The objective of the experiment was to determine the RBV of MHA-Ca compared with DL-Met to support growth performance of 11 to 20 kg pigs. A total of 504 piglets [DNA x Pietrain pigs; approximately 42 d of age and 11 kg body weight (BW)] were used in two blocks of 252 pigs each with 6 pigs (3 entire males and 3 gilts) per pen and 12 replicate pens per treatment. Within each block, pigs were randomly allotted to 7 diets for 21 d. A basal diet (BD) was formulated to be adequate in all essential nutrients except for Met (0.22% standardized ileal digestible Met), which was 61% of the SID Met requirement of 0.36% recommended by NRC (2012) for 10 to 25 kg pigs. The Met-deficient BD was then supplemented with 3 graded levels of DL-Met (0.05, 0.10, and 0.15%) or MHA-Ca (0.077, 0.154, and 0.231%) at the expense of corn. Pigs had ad libitum access to feed (mash) and water. Individual pigs and pen feed disappearance were weighed on d 0 and d 21 to calculate average daily gain (ADG), average daily feed intake (ADFI) and feed conversion rate (FCR). On d 21, blood samples were collected from 1 pig per pen after 8 h fasting to determine total protein, albumin and glucose. Data were analyzed as a randomized complete block design using GLM procedure of SAS 9.4 (SAS Inst. Inc.). Orthogonal-polynomial contrasts were used to determine linear and quadratic effects of increasing levels of DL-Met and MHA-Ca, and the effect of Met sources. The final BW at d 21 and overall average daily gain (ADG; 375, 401, 429, 430, 402, 431, and 428 g/d for diets 1 to 7, respectively) increased linearly (P < 0.01) by graded additions with both Met sources. The ADFI was not different but the overall FCR (1.64, 1.55, 1.51, 1.45, 1.59, 1.46, and 1.50 for diets 1 to 7, respectively) improved linearly (P < 0.01) by supplementation with DL-Met or MHA-Ca. Pig performance responses (ADG, ADFI and FCR) was not different (P ≥ 0.15) when Met was supplemented in the Met-deficient diet at a DL-Met to MHA-Ca ratio of 65:100 on a product basis. Plasma concentrations of total protein, albumin and glucose on d 21 were not affected (P >0.05) by dietary treatments or Met sources. Based on ADG as a response of Met supplementation level, the linear and exponential regressions estimated the RBV of 64 and 65%, respectively for MHA-Ca relative to DL-Met (product-to-product basis) in 11 to 20 kg pigs.

178 Effects of L-glutamate and L-aspartate supplementation on systemic and intestinal immune responses of weaned piglets challenged with F18 enterotoxigenic Escherichia coli challenge

Abstract L-glutamate (Glu) and L-aspartate (Asp) are pivotal components in the cellular metabolic pathways and immune regulation of pigs. This study aimed to investigate the effects of Glu and Asp on systemic and intestinal immune responses of weaned piglets challenged with an enterotoxigenic Escherichia coli (ETEC) F18. A total of 49 weaned pigs [initial body weight (BW) = 8.18 ± 1.54 kg] were randomly assigned to one of seven treatments, with seven piglets per treatment. The treatments included negative control (NC) and positive control (PC) that were fed with control diet without or with F18 enterotoxigenic Escherichia coli (ETEC) challenge, respectively. The other five dietary treatments were supplemented with either 1% or 2% Glu or Asp, or 50 mg/kg of Carbadox, and were challenged with ETEC. All pigs except for NC were challenged with F18 ETEC orally for three consecutive days at the dose of 1010 CFU•dose-1•d-1. The study period lasted for 3 wk, with a 7-d adaptation period and 14 d post-inoculation (PI). Blood samples were collected on d 0 before ETEC inoculation, and on d 2, 5, and 14 PI to analyze systemic inflammatory markers by the enzyme-linked immunosorbent assays. On d 14 PI, intestinal mucosa from jejunum and ileum were collected to analyze the mRNA expression of inflammatory cytokines by quantitative polymerase chain reaction. All data were analyzed with ANOVA using the PROC MIXED of SAS with pig as the experimental unit. Among all treatments, pigs fed with carbadox had the least (P < 0.05) serum TNF-α, haptoglobin, and C-reactive protein throughout the experiment. Pigs supplemented with 1% Glu had similar concentrations of TNF-α, haptoglobin, and C-reactive protein, while pigs fed with 2% Asp had similar serum haptoglobin compared with pigs fed with carbadox. Compared with PC, supplementing Glu and Asp reduced (P < 0.05) the mRNA expression of IL12 in jejunal mucosa. Pigs fed with carbadox tended to have greater (P < 0.10) expression of IFNG in ileal mucosa than pigs in PC, while no difference was observed in IFNG expression among 1% Glu, 1% or 2% Asp, and carbadox groups. In addition, pigs fed with 1% Glu or 2% Asp had greater (P < 0.05) expression of IL12 in ileal mucosa than pigs in PC. However, dietary treatments did not affect the gene expression of TNFA, IL1B and TGFB in jejunal and ileal mucosa of pigs. In conclusion, results of the present study indicate that dietary supplementation of 1% Glu or 2% Asp affected both systemic and intestinal immune responses of ETEC challenged weaned pigs with similar trend as the supplementation of carbadox.

149 Development of a leaky gut model: Impact of 24-h feed restriction on ileal expression of tight junction proteins in weaned pigs

Abstract Weanling barrows, 26 to 28 d old [n = 90; Duroc terminal line Fast genetics, initial body weight (BW)= 5.63 ± 0.43 kg] were used to evaluate the effect of 24-h complete feed restriction on growth and intestinal integrity. Upon arrival at the NutriQuest modeling center (Fergus Falls, MN), pigs were weighed, blocked by BW, and randomly assigned to 1 of 30 pens. Five treatments were randomly assigned to 6 pens: 1) Positive Control (PC), and 4 treatments that had 24-hr feed restriction (FR); 2) Negative control (NC), 3) Direct Fed Microbial (DFM) A, 4) DFM B, and 5) DFM C. Feed intake and BW were determined weekly and before and after FR. All pigs were fed a common diet. Fifteen pigs were removed prior to FR due to enteric disease. On d 18, feed was removed from pigs on treatments 2 to 5 for 24 h. After 24 h, 8 pigs per treatment were harvested for tissue collection. Feed access was restored to the remaining pigs who were harvested 3 d later. Ileal samples were collected and immediately frozen. Plasma and fecal samples were collected upon arrival, at FR, and 24 h and 4-d post-FR. Ileal expression of claudin-3, occludin, and tight-junction-1 genes was determined using QuantStudio 3 Real Time PCR system. Fold-change in gene expression compared with NC was calculated for each treatment. Plasma samples were analyzed for LPS binding protein (LBP) concentration, and fecal samples were analyzed for myeloperoxidase (MPO) concentration. Pig (n = 75) was the experimental unit for BW, LBP, MPO, and gene expression data. Pen (n = 30) was the experimental unit for feed intake. Data were analyzed using PROC GLIMMIX of SAS. Preplanned orthogonal contrasts were performed to determine the effect of PC vs. 4 FR treatments. To determine the relationship of tight junction gene expression and LBP and MPO, PROC REG of SAS was used. Data are described as PC vs. FR treatments to focus on model development. Final BW for pigs on PC was greater compared with the FR treatments (P < 0.05; Table 1). From 1 to 4 d post-FR, average daily gain (ADG) for FR treatments was greater than PC (P < 0.01). There was no effect of treatment on feed intake or feed efficiency. Plasma LBP and fecal MPO concentrations were greater 24 h post-FR for pigs on PC compared with FR treatments (P < 0.01). Fold change in occludin and claudin-3 expression was greater for PC compared with FR treatments (P ≤ 0.05). For pigs euthanized 24 h post-FR, there was a positive relationship between LBP concentrations and occludin expression (P < 0.02). Twenty-four hour FR in weaned pigs alters ileal tight junction gene expression and plasma LBP concentrations; therefore, a timed FR event may serve as a model for leaky gut.

PSV-9 Effects of increasing amounts of high-oleic or conventional soybeans in diets fed to mid-lactation dairy cows on measures of dry matter intake and lactation

Abstract The objective of the study was to determine effects of increasing amounts of conventional (CONV) or high-oleic (HO) soybean in diets fed to mid-lactation dairy cows on dry matter intake (DMI) and performance. Rumen and duodenally cannulated cows (n = 8) were split into two groups and placed in a split-plot design. The whole plot factor was type of soybean (CONV vs. HO) and was provided to cows in a crossover arrangement so that each cow was provided both types of soybeans throughout the experiment. The subplot factor was amount of soybeans (0, 4, 8, 12%). Each group of cows receiving the same type of soybean were placed in a 4 x 4 Latin square with treatment sequences balanced for carryover effects. Diets were formulated to contain 18.5% crude protein and to slightly exceed requirements for ruminally degradable protein. Additions of soybeans replaced soybean meal and soybean hulls so that diets provided similar amounts of soy-based protein and fiber. During the last 4 d of each 14-d period, spot samples of feed, orts, and milk were collected together with measures of milk production and feed intake. Data were analyzed using the MIXED procedure of SAS. There were no interactions of soybean type and amount on lactation or DMI except for measures of milkfat, milk energy concentration and milkfat yield. Increasing amounts of HO resulted in a quadratic response in milkfat and energy concentration together with measures of milkfat yield. Specifically, these measures were greater (P ≤ 0.02) when cows were fed diets containing 4 and 8% HO compared with diets containing 0 or 12% HO; increasing amounts of CONV had no impact (P ≥ 0.32) on milkfat and energy concentration or milkfat yield. Measures of milk yield were not affected by type or amount of soybean (P ≥ 0.54), but measures of energy corrected milk yield (ECM) tended to respond differently to increasing amounts of soybean from either CONV or HO (P = 0.07). Interestingly, ECM responded quadratically (P = 0.02) to increasing amounts of HO but not to increasing amounts of CONV (P ≥ 0.58). Overall, however, ECM tended (P = 0.06) to be slightly greater with HO compared with CONV (42.5 vs. 41.1 kg/d). Yet, DMI was also greater (P ≤ 0.01) with HO compared with CONV (26.6 vs. 24.4 kg/d). Differences in DMI, therefore, resulted in a tendency (P = 0.07) for greater efficiency of ECM production when cows were fed CONV vs. HO. Overall, treatments had no effect on measures of milk protein, urea-nitrogen, solids-not-fat or lactose concentrations or yields. These data seem to indicate that performance responses among lactating cows to increasing amounts of HO are dynamic but that responses to CONV are not.

150 Development of a leaky gut model: Meta-analysis of plasma and fecal markers of leaky gut

Abstract Intestinal integrity can be compromised in livestock and poultry species during stress events, including off-feed events. To develop a leaky gut model, weanling barrows 26 to 28 d old [n = 307; Duroc terminal line Fast genetics, initial body weight (BW) = 5.90 ± 0.47 kg), were used in a total of 6 randomized complete block experiments at the NutriQuest modeling center (Fergus Falls, MN). All trials included positive control (PC) and negative control (NC) treatments. The other treatments evaluated (n = 8) were various strains and levels of direct-fed microbial (DFM) products. With the exception of PC, all treatments had complete 24-h feed restriction (FR) approximately 18 d after arrival. Pigs assigned to PC had ad libitum access to feed. Pigs were housed 3 pigs per pen in 1 of 30 pens, and all pigs were fed a common high-quality basal diet. For 3 trials, pigs received Excede (Zoetis; Parsipanny, NJ) IM upon arrival and were provided gentamycin via water for 3 d post-arrival. For 3 trials, no prophylactic antibiotics were administered. Eighteen days post-arrival, fecal and plasma samples were collected from all pigs and feed was removed from FR pens. After 24 h, plasma and fecal samples were collected from all pigs, and one-half of the pigs per trial were harvested for tissue collection. Four days after FR, samples were collected similarly, and the remaining pigs were harvested. Plasma samples were analyzed for LPS binding protein concentrations (LBP), and fecal samples were analyzed for myeloperoxidase (MPO) concentrations. Data from the PC (n = 90) and NC (n = 92) treatments only were analyzed as Individual Participant Data meta-analysis using PROC MIXED of SAS. Data from all pigs (n = 307) were analyzed using PROC REG of SAS with forward step-wise selection to determine relationships between response variables. Plasma LBP concentrations did not differ between PC and NC 24 h after FR (P = 0.14). However, change in LBP concentrations and the ratio of LBP immediately before and after FR, differed between PC and NC (P < 0.001). For NC, LBP concentrations decreased, and for PC, LBP concentrations increased (Table 1). There was no effect of treatment on fecal MPO concentrations. Using the broader dataset (n = 307), variables that were predictive of plasma LBP concentrations immediately following FR were trial (P < 0.001), treatment (P < 0.001), and fecal MPO concentrations (P < 0.05). Variables that were predictive of change in LBP concentrations before and after FR were trial (P < 0.001), treatment (P < 0.001), and initial BW (P < 0.01). These data, taken with data presented in a companion abstract on ileal tight junction gene expression, suggest that plasma LBP concentrations immediately following a FR event are a useful marker of leaky gut. Fecal MPO concentrations do not consistently respond to FR.

139 Effect of dietary crude protein concentration and sensory feed additive on nursery pig growth performance

Abstract Maximizing growth performance and feed intake for nursery pig diets is an ongoing challenge in the US swine industry. Using sensory feed additives is one strategy to optimize feed intake and growth rates of pigs. Weanling pigs (n = 416; average age = 19.8 d) were used to test the effects of sensory feed additive (SA; Luctamax Piglet, Lucta, Mahwah, NJ) addition to control (CON) and low crude protein (LCP; decreased 4% CP) nursery diets in a 2 x 2 factorial design (CON, CON+SA, LCP, LCP+SA). Decreased dietary CP was achieved by removing or decreasing fish meal and soy concentrate in phases 1 and 2 and reducing soybean meal in phase 3 and adding synthetic amino acids to maintain amino acid ratios (NRC 2012). Nursery treatments were fed in three phases; phase 1 (PH1, d 0-7, 0.10% SA), phase 2 (PH2, d 7-21, 0.10% SA), and phase 3 (PH3, d 21-36, 0.05% SA). Pigs were blocked by initial body weight (BW), sex, ancestry, and housed at 6 or 7 pigs/pen with 15 pens/treatment. Weekly pig BW and feed intake were used to calculate pen average daily gain (ADG), average daily feed intake (ADFI), and gain to feed ratio (G:F). Data were analyzed using GLM procedure of SAS 9.4. During PH1 pigs receiving the CON diets tended (P = 0.065) to have a greater G:F compared with pigs receiving LCP diets. For the remainder of this study there were consistent interactions between dietary CP and SA. When SA was added to the LCP diet ADFI increased but when SA was added to CON diet ADFI decreased and these changes in feed intake often led to similar dietary interactions in ADG. During PH2, pigs fed CON or LCP+SA diets had greater ADFI (P = 0.032) and tended to have a greater ADG (P = 0.094) but reduced G:F (P = 0.100) compared with pigs fed CON+SA or LCP diets. In PH3, pigs fed CON or LCP+SA diets had a greater ADG (P = 0.027) and ADFI (P = 0.011) compared with CON+SA or LCP diets. Pigs fed LCP diets had greater ADFI (P = 0.001) but reduced G:F (P = 0.0001) than pigs fed CON diets. Overall, pigs fed CON or LCP+SA diets had greater ADFI (P = 0.016) and tended to have a greater ADG (P = 0.059) but reduced G:F (P = 0.067) compared with pigs fed CON+SA or LCP diets. Pigs fed CON diets had a greater G:F (P < 0.001) compared with pigs fed LCP diets, while LCP pigs tended to have a greater ADFI (P = 0.054) compared with CON pigs. Inclusion of SA to LCP diet mitigated the adverse impact of replacing whole dietary proteins with high inclusion of synthetic amino acids. Careful consideration is needed when using sensory feed additives so as not to impede growth performance in diets containing traditionally greater levels of crude protein.

64 Effect of feeding intact protein from soybean meal instead of crystalline amino acids on growth performance, protein synthesis, and immune response of growing pigs

Abstract The objective was to test the hypothesis that feeding intact protein from soybean meal (SBM) instead of crystalline amino acids (AA) does not affect growth performance, protein synthesis, intestinal morphology, or immune response of growing pigs. A control corn-SBM diet was used. Three additional diets were formulated by reducing the inclusion rate of SBM and adding 3, 4, or 5 crystalline AA (i.e., Lys, Met, Thr, Trp, Val) to the diets, which resulted in reducing the protein concentration from 20.0% to 13.4%. All diets were formulated to meet requirements for standardized ileal digestible indispensable AA. A total of 176 pigs [initial body weight (BW) = 32.2 kg; SD = 4.2] were used. On d 1, 16 randomly chosen pigs were euthanized and body nutrient composition was determined. The remaining 160 pigs were allotted to the 4 diets using a randomized complete block design with 4 pigs per pen and 10 replicate pens per diet. Starting BW was the blocking factor. Individual pig BW and feed consumption were recorded. On d 28, one pig per pen was slaughtered and blood, carcass, and viscera were collected and analyzed for N, fat, and energy to calculate deposition in the body. Ileal mucosa, ileum and colon tissue and digesta were analyzed for blood characteristics, cytokines, tissue morphology, ammonia, and gene expression. Contrasts coefficients were used to determine linear and quadratic effects of reducing dietary protein. Results indicated that average daily gain, average daily feed intake, gain to feed ratio, carcass characteristics, and protein, lipid, and energy depositions were not affected by reducing SBM and increasing crystalline AA in diets (Table 1). However, energy efficiency tended to decrease (quadratic, P = 0.083) by reducing dietary protein. Blood urea N was reduced (linear, P < 0.001) as dietary protein was reduced, but blood total protein was not affected by dietary treatment. Cytokines, ileal and jejunal morphologies, and gene abundance of AA transporters in the ileal mucosa were not affected by dietary protein. Ammonia concentrations in ileal digesta increased and then decreased (quadratic, P = 0.043) and colon digesta tended to increase and then decrease (quadratic, P = 0.074) as dietary protein was reduced. Bacteria protein in colon digesta was reduced (linear, P = 0.030) by reducing dietary protein. In conclusion, providing AA from intact protein from SBM or from crystalline AA did not affect growth performance, intestinal morphology, and immune response of pigs. However, SBM in diets can provide energy that may be utilized more efficiently than energy from diets with less SBM and crystalline AA. Further research is suggested to elucidate if SBM improves utilization of other nutrients in diets for pigs.

Greater difference between airborne and flower pollen chemistry, than between pollen collected across a pollution gradient in the Netherlands

The prevalence in allergic diseases has increased considerably in the past decades. An important trigger of the symptoms of allergic rhinitis (hay fever) is the pollen of wind-pollinating plants. This pollen is developed by plants and is released into the air where it gets exposed to environmental influences and air pollution. We investigated the chemical changes to pollen that occur after release from the flower in a rural (Veluwe) and an urban (Amsterdam) site in the Netherlands using Fourier Transform Infrared (FTIR) spectroscopy. During the spring/summer of 2020 (during the COVID pandemic) the pollen of nine taxa (Alnus, Betula, Fagus, Fraxinus, Pinus, Plantago, Poaceae, Quercus and Salix) were collected directly from flowers and the air (using a mobile sampler). FTIR spectra were obtained for multiple individual pollen grains for each taxa. The spectra obtained from airborne pollen collected at the rural vs. urban sites did not show any statistical difference. This is possibly a result of a reduced difference in pollutant concentrations between the two sites due to the COVID-19-lockdown measures were in place. However, consistent differences in the FTIR spectra recovered from airborne vs. flower pollen were recorded for all pollen taxa. After the release from the flower the chemical composition of the pollen changed: (i) polysaccharides are converted to monosaccharides; (ii) protein concentration and/or nitration/oxidation level is altered; (iii) lipids are modified and/or reduced in concentration. These changes may alter the allergenicity of the pollen and suggest that further work on the allergenic nature of airborne pollen is required.

Longitudinal Fluctuations in Protein Concentrations and Higher-Order Structures in the Plasma Proteome of Kidney Failure Patients Subjected to a Kidney Transplant.

Using proteomics and complexome profiling, we evaluated in a year-long study longitudinal variations in the plasma proteome of kidney failure patients, prior to and after a kidney transplantation. The post-transplant period was complicated by bacterial infections, resulting in dramatic changes in the proteome, attributed to an acute phase response (APR). As positive acute phase proteins (APPs), being elevated upon inflammation, we observed the well-described C-reactive protein and Serum Amyloid A (SAA), but also Fibrinogen, Haptoglobin, Leucine-rich alpha-2-glycoprotein, Lipopolysaccharide-binding protein, Alpha-1-antitrypsin, Alpha-1-antichymotrypsin, S100, and CD14. As negative APPs, being downregulated upon inflammation, we identified the well-documented Serotransferrin and Transthyretin, but added Kallistatin, Heparin cofactor 2, and interalpha-trypsin inhibitor heavy chain H1 and H2 (ITIH1, ITIH2). For the patient with the most severe APR, we performed plasma complexome profiling by SEC-LC-MS on all longitudinal samples. We observed that several plasma proteins displaying alike concentration patterns coelute and form macromolecular complexes. By complexome profiling, we expose how SAA1 and SAA2 become incorporated into high-density lipid particles, replacing largely Apolipoprotein (APO)A1 and APOA4. Overall, our data highlight that the combination of in-depth longitudinal plasma proteome and complexome profiling can shed further light on correlated variations in the abundance of several plasma proteins upon inflammatory events.

Effects of a blend of acidifiers added continuously to the drinking water of nursery piglets

Context Adding organic and inorganic acids to the drinking water is a potential alternative to improve production results in nursery piglets, and a strategy for reducing the use of antimicrobials. Aims To assess the effects of adding a blend of organic and inorganic acids to drinking water on the oxireduction potential (ORP) of water, and growth performance, biochemical, antioxidant and blood gas profiles, intestinal morphometry, biometry of organs, and intestinal microbiome of nursery piglets. Methods Experiment involved 1080 female piglets assessed over 30 days (Landrace × Large White, weaned at 26 days of age, weighing 6.8 ± 0.29 kg), randomly assigned to the following three treatments: (1) piglets received drinking water of natural pH of 7.39 (pH7.39), (2) piglets received drinking water of pH of 5.31 (pH5.31), and (3) piglets received drinking water of pH of 3.40 (pH3.40), with eight replicates and 45 piglets/stall. Piglets had ad libitum access to diet and water throughout the experiment, and received different diets in each phase. Key results Receiving drinking water of pH3.40 promoted a higher ORP, and a lower total water consumption was observed in Starter II piglets with pH5.31. There was no treatment effect on the performance, but mortality was lower in starter piglets in the pH3.40 treatment. Animals in the pH3.40 treatment had lower blood alanine aminotransferase and aspartate aminotransferase activities, differing from the result obtained for total cholesterol concentrations. Animals in the pH3.40 treatment showed a lower concentration of total protein than did piglets in the pH7.39 treatment. Piglets in the pH3.40 treatment had higher weights of empty stomach, spleen, and liver with gallbladder than did those in the pH7.39 treatment. Animals in the pH3.40 treatment showed a lower relative abundance of the Escherichia–Shigella genus and bacteria of Enterobacteriaceae family than did those in the pH5.31 treatment. Conclusions Acidification of drinking water to pH3.40 for nursery piglets can be used to partially increase the ORP of the water, and promote a greater weight of organs with metabolic and immune functions, while decreasing mortality. In addition, acidification of drinking water does not compromise performance or intestinal histology, but improves the profile of the intestinal microbiome in piglets. Implications This research has provided new information on the effects of acidifiers in the drinking water for nursery piglets and has shown their potential use for bolstering general health.

Concentrations of selected immunological parameters in the serum and processing fluid of suckling piglets and the serum and colostrum of their mothers

BackgroundBlood sampling from neonatal piglets is related to multiple disadvantages. Therefore, a new, alternative matrix is required to assess piglets’ early immune status efficiently. The present study aimed to assess the usefulness of processing fluid for determining selected piglets’ immune parameters. 264 pigs − 31 sows, 146 male piglets, and 87 female piglets from commercial indoor farrow-to-finish pig herd were included in this study. 264 serum, 31 colostrum, and 146 processing fluid samples were collected. Serum was collected from all animals, colostrum was collected from sows, and processing fluid was collected from male piglets only. Using commercial ELISA tests, the concentration of various immunoglobulins, cytokines, and acute phase proteins was assessed in each matrix. Statistical analyses were employed to determine differences in the concentration of measured indices between piglets’ serum and processing fluid and correlations in the concentration of tested indices between particular sets of matrices.ResultsStatistical analyses did not reveal significant differences in the IgG, IgA, IL-1β, IL-4, IL-6, and IFN-γ concentration between piglets’ serum and processing fluid (p > 0.05). A positive correlation (p < 0.05) regarding the concentration of some indices between processing fluid and samples collected from sows was also observed.ConclusionsProcessing fluid can be considered a promising alternative to blood for assessing some immunological indices in piglets, such as IgG, IgA, IL-1β, IL-4, IL-6, and IFN-γ, and, possibly, in the indirect assessment of some indices in lactating sows, including IgA, IL-1β, IL-4, IL-6, IL-8, IFN-γ, or Pig-MAP.

Toxicological Effects of Commonly Used Mosquito Coil Smoke on Liver and Lung Function in Rats

The utilization of mosquito coils as a household-level strategy for mosquito control is widely practiced in malaria-endemic regions spanning Africa, Asia, and South America. However, the continuous application of mosquito coils has prompted significant apprehensions concerning environmental and health implications. This research aims to evaluate the potential hepatic and pulmonary toxicities associated with the inhalation of mosquito coil smoke in rat subjects over both sub-chronic and chronic durations. A questionnaire was used to make inquiries regarding the prevalent types and brands of mosquito coils employed, the manner in which they are used, the durations for which they are utilized. A total of twenty-four albino rats were categorized into three distinct groups: the control group and two exposed groups. These groups were subjected to distinct brands of mosquito coil smoke across two distinct exposure regimens – a sub-chronic exposure lasting 4 weeks and a chronic exposure spanning 12 weeks. Following the exposure periods, blood samples were collected for the analysis of serum-based and biochemical markers, including serum aspartate aminotransferase (AST), alanine aminotransferase (AST), alkaline phosphatase (ALP), gamma-glutamyl transferase (GGT), bilirubin, serum bicarbonate, and Lactate dehydrogenase (LDH), oxygen saturation, C-reactive protein, total protein, and microscopic analysis of lung tissue and liver tissue. The histopathological analysis of lung tissue was appropriately included. However, the analysis of liver tissue was omitted. The results demonstrated a significant elevation in the levels of these biochemical markers among the exposed rats in comparison to the control group. Concurrently, a decrease in blood oxygen saturation was noted in the exposed rats when contrasted with the control group. Conversely, the serum concentrations of high-sensitivity C-reactive protein, total protein, and alanine aminotransferase (ALT) did not exhibit notable differences in relation to the control group. Additionally, microscopic analysis of lung tissue indicated the presence of histopathological lesions in the exposed rats, suggesting a detrimental impact of mosquito coil smoke exposure in a time-dependent progression.

Interaction of obesity and proteins associated with the NLRP3 inflammasome following mild traumatic brain injury

The NOD-like receptor pyrin domain-containing protein 3 (NLRP3) inflammasome has been associated with worse outcomes from severe traumatic brain injury (TBI). The NLRP3 inflammasome is also strongly associated with other pro-inflammatory conditions, such as obesity. Little is known about the potential effect of mild TBI (mTBI) on the NLRP3 inflammasome and the extent to which modifying factors, such as obesity, may augment the inflammatory response to mTBI. The purpose of this study was to evaluate the association of NLRP3 inflammasome proteins with obese body mass index (BMI ≥ 30) within 24 h of mTBI after presenting to a level 1 trauma center emergency department. This is a secondary analysis of prospectively enrolled patients with mTBI who presented to the emergency department of one U.S. Level 1 trauma center from 2013 to 2018 (n = 243). A series of regression models were built to evaluate the association of NLRP3 proteins obtained from blood plasma within 24 h of injury and BMI as well as the potential interaction effect of higher BMI with NLRP3 proteins (n = 243). A logistic regression model revealed a significant association between IL-18 (p < 0.001) in mTBI patients with obese BMI compared to mTBI patients with non-obese BMI (< 30). Moderation analyses revealed statistically significant interaction effects between apoptotic speck-like protein (ASC), caspase-1, IL-18, IL-1β and obese BMI which worsened symptom burden, quality of life, and physical function at 2 weeks and 6 months post-injury. Higher acute concentrations of IL-1β in the overall cohort predicted higher symptoms at 6-months and worse physical function at 2-weeks and 6-months. Higher acute concentrations of IL-18 in the overall cohort predicted worse physical function at 6-months. In this single center mTBI cohort, obese BMI interacted with higher acute concentrations of NLRP3 inflammasome proteins and worsened short- and long-term clinical outcomes.