Two experiments were conducted to investigate the production effects of N-acetyl-l-methionine (NALM; experiment 1) and to estimate its bioavailability (BA) and rumen escape (RE; experiment 2), respectively, in lactating dairy cows. In experiment 1, 18 multiparous Holstein cows were used in a replicated, 3 × 3 Latin square design experiment with three 28-d periods. Treatments were (1) basal diet estimated to supply 45 g/d digestible Met (dMet) or 1.47% of metabolizable protein (MP; control), (2) basal diet top-dressed with 32 g/d of NALM to achieve dMet supply of 2.2% of MP, and (3) basal diet top-dressed with 56 g/d of NALM to achieve dMet supply of 2.6% of MP. The NALM treatments supplied estimated 17 and 29 g/d dMet from NALM, respectively, based on manufacturer's specifications. In experiment 2, 4 rumen-cannulated lactating Holstein cows were used in a 4 × 4 Latin square design experiment with four 12-d periods. A 12-d period for baseline data collection and 4 d for determination of RE of NALM preceded the Latin square experiment. For determination of RE, 30 g of NALM were dosed into the rumen simultaneously with Cr-EDTA (used as a rumen fluid kinetics marker) and samples of ruminal contents were collected at 0 (before dosing), 1, 2, 4, 6, 8, 10, 14, 18, and 24 h after dosing. Rumen escape of NALM was calculated using the estimated passage rate based on the measured Cr rate of disappearance. Bioavailability of abomasally dosed NALM was determined using the area under the curve of plasma Met concentration technique. Two doses of l-Met (providing 7.5 and 15 g of dMet) and 2 doses of NALM (11.2 and 14.4 g of dMet) were separately pulse-dosed into the abomasum of the cows and blood was collected from the jugular vein for Met concentration analysis at 0 (before dosing), 1, 2, 4, 6, 8, 10, 12, 14, 18, and 24 h after dosing. Supplementation of NALM did not affect DMI, milk yield, feed efficiency, or milk protein and lactose concentrations and yields in experiment 1. Milk fat concentration and energy-corrected milk yield decreased linearly with NALM dose. Plasma Met concentration was not affected by NALM dose. The estimated relative BA of abomasally dosed NALM (experiment 2) was 50% when dosed at 14.4 g/cow (11.2 g/d dMet from NALM) and 24% when dosed at 28.8 g/cow (14.4 g/d dMet from NALM). The estimated RE of NALM was 19% based on the measured kp of Cr at 11%/h. The total availability of ingested NALM was estimated at 9.5% for the lower NALM dose when taking into account RE (19%) and bioavailability in the small intestine (50%). Overall, NALM supplementation to mid-lactation dairy cows fed a MP-adequate basal diet below NRC (2001) recommendations (45 g/d or 1.47% Met of MP) decreased milk fat and energy-corrected milk yields but did not affect milk or milk true protein yields. Further evaluation of BA of NALM at different doses is warranted. In addition, intestinal conversion of NALM to Met needs additional investigation to establish a possible saturation of the enzyme aminoacylase I at higher NALM doses.

The present study investigated the plasma methionine (Met) and residual potential of N-acetyl-L-Met (NALM) in lactating dairy cows. Six cows (75 ± 20.1 days-in-milk) were used in a replicated 3 × 3 Latin square design. Within each square, cows were randomly assigned to a sequence of three dietary treatments during each of the three 13-day periods (10 days of treatment adaptation and 3 days of data collection and sampling). The three dietary treatments are as follows: basal diet without NALM (control); control diet with 30 g/day of NALM by rumen placement (30NALM), and control diet with 60 g/day of NALM by rumen placement (60NALM). Rumen NALM dosing led to a linear increase in plasma Met concentration. Abomasal infusion with NALM resulted in both linear and quadratic increases in plasma Met concentration. No NALM was detected in milk, liver, plasma, and muscle samples after rumen placement or abomasal infusion. Supplementation of NALM did not affect dry matter intake and milk yield. The absence of plasma NALM and increases in plasma Met concentration for both ruminal and abomasal NALM dosing suggest that NALM supplemented by either rumen placement or abomasal infusion to lactating dairy cows is deacetylated before entering the central circulation.

Objective: We investigated the lactational performance and N utilization of dairy cows fed different forms of Lys supplements with the main focus on Lys derivatives of Nacetyl-L-Lys (NALL) during mid to late lactation. Materials and Methods: Forty lactating Holstein dairy cows (151 +/- 78.0 DIM) were assigned to 1 of 4 treatments (n = 10) in a randomized complete block design. The 4 dietary treatments were as follows: (1) TMR without Lys supplementation (control), (2) TMR with N epsilon-acetyl-L-Lys supplementation at 51.5 g/d (epsilon NALLT), (3) TMR with N alpha, epsilon-acetyl-L-Lys supplementation at 63 g/d (diNALLT), and (4) TMR with commercial rumen-protected Lys (AjiPro-L, Ajinomoto Heartland Inc.) supplementation at 100 g/d (CRPLT). Results and Discussion: Feeding of diNALLT and the CRPLT decreased DMI. Milk yield, component yield, and component concentrations were not affected by Lys supplementation. In contrast, feed conversion efficiency (milk yield/DMI) was greatest for diNALLT. The efficiencies of 3.5% FCM yield/DMI and ECM yield/DMI did not differ among treatments. Feeding the diNALLT and the CRPLT tended to reduce manure N excretion compared with the control and the epsilon NALLT. Overall, results from the current study suggest that supplementing a Lys derivative, diNALL, to mid- to late-lactation dairy cows resulted in similar production performance relative to a commercial rumen-protected Lys product. Implications and Applications: A sizable increase in feed conversion efficiency (milk yield/DMI) and a trend for a reduction in manure N excretion when cows were supplemented with diNALL resulted from its effect to reduce DMI and N intake. This result indicates that the developmental diNALL product is metabolically active in lactating dairy cows. Further investigation is warranted to understand the biological availabilities of NALL in lactating dairy cows.

Probiotics have received attention as alternatives to antibiotics due to their relative biosafety. Despite the health-promoting effects of multispecies probiotic formulation (MPF) in weaning pigs were well-documented, the health-promoting potential of MPF in growing-finishing pigs remain limited. Therefore, we conducted a six-week experiment to investigate the beneficial effects of MPF supplementation (the formulations containing Lactobacillus plantarum CJLP243, L. fermentum LF21, L. salivarius E4101, Leuconostoc paramesenteroides KJP421, Bacillus subtilis CJMPB957 and B. licheniformis CJMPB283) on intestinal environments and hepatic lipid metabolism in growing-finishing pigs. The results showed that the growth performance of MPF group was improved compared to control group during the experimental period without affecting feed intake. The dietary MPF supplementation also significantly increased the jejunal mRNA expression levels of zonula occludens-1 (ZO-1) and decreased that of interleukin-12 (IL-12). Dietary MPF supplementation was found to not only reduce the hepatic triglyceride concentration but down-regulate hepatic mRNA expression levels of lipid metabolism-related markers, such as as sterol regulatory element-binding transcription factor 1c (SREBP-1c), CCAAT/enhancer binding protein ? (CEBP?), acetyl coA carboxylase (ACC) and carnitine palmitoyl transferase 1? (CPT1?). Nonetheless, MPF supplementation also had the capability to modulate gut microbiota populations, including Clostridiaceae, Lachnospiraceae, and Turicibacter. Collectively, our results suggest that dietary MPF inclusion could be a promising approach to promote the growth performance and the overall health of growing-finishing pigs via modulation of gut microbiota.

The objective of this experiment was to evaluate the effects of supplementing a rumen-protected source of Met, N-acetyl-l-methionine (NALM), on lactational performance and nitrogen metabolism in early- to mid-lactation dairy cows. Sixty multiparous Holstein dairy cows in early lactation (27 ± 4.3 d in milk, SD) were assigned to 4 treatments in a randomized complete block design. Cows were blocked by actual milk yield. Treatments were as follows: (1) no NALM (control); (2) 15 g/d of NALM (NALM15); (3) 30 g/d of NALM (NALM30); and (4) 45 g/d of NALM (NALM45). Diets were formulated using a Cornell Net Carbohydrate and Protein System (CNCPS) v.6.5 model software to meet or exceed nutritional requirements of lactating dairy cows producing 42 kg/d of milk and to undersupply metabolizable Met (control) or supply incremental amounts of NALM. The digestible Met (dMet) supply for control, NALM15, NALM30, and NALM45 were 54.7, 59.8, 64.7, and 72.2 g/d, respectively. The supply of dMet was 88, 94, 104, and 115% of dMet requirement for control, NALM15, NALM30, and NALM45, respectively. Milk yield data were collected, dry matter intake (DMI) was measured daily, and milk samples were collected twice per week for 22 wk. Blood, ruminal fluid, urine, and fecal samples were collected during the covariate period and during wk 4, 8, and 16. Data were analyzed using the GLIMMIX procedure of SAS (SAS Institute) using covariates in the model for all variables except body weight. Linear, quadratic, and cubic contrasts were also tested. Treatments did not affect DMI, milk yield, and milk component concentration and yield; however, feed efficiency expressed as milk yield per DMI and 3.5% fat-corrected milk per DMI were quadratically affected, with greater response observed for NALM15 and NALM30 compared with control. Acetate proportion linearly increased, whereas propionate proportion linearly decreased with NALM supplementation. Blood urea nitrogen linearly decreased with NALM supplementation. Total plasma essential AA concentrations were quadratically affected, as greater values were observed for control and NALM45 than other treatments. Plasma Met concentration was quadratically affected as lower levels were observed with NALM15, whereas Met concentrations increased with NALM45 compared with control. Nitrogen utilization efficiency and apparent total-tract nutrient digestibility were not affected by treatment. Supplementation of NALM at 15 or 30 g/head per day resulted in the greatest improvements in feed efficiency without affecting N metabolism of early- to mid-lactation dairy cows.

Gastrointestinal microbiota impact host's biological activities, including digestion of indigestible feed components, energy harvest, and immunity. In this study, fecal microbiota of high body weight (HW) and low body weight (LW) growing pigs at 103 days of age were compared. Principal coordinates analysis separated the HW and LW groups into two clusters, indicating their potential differences between microbial community composition. Although the abundances of two major phyla, Firmicutes and Bacteroidetes, did not significantly differ between the HW and LW groups, some genera showed significant differences. Among them, Peptococcus and Eubacterium exhibited strong positive correlations with body weight (BW) and average daily gain (ADG) (Rho > 0.40), whereas Treponema, Desulfovibrio, Parabacteroides, and Ruminococcaceae_unclassified exhibited strong negative correlations with BW and ADG (Rho < -0.40). Based on these results, the structure of intestinal microbiota may affect growth traits in pigs through host-microbe interactions. Further in-depth studies will provide insights into how best to reshape host-microbe interactions in pigs and other animals as well.

High nitrogen (N) fertilizer costs and increased environmental stewardship have renewed interest in grass-legume pastures. This research compared herbage mass, nutritive value, steer growth performance and economics of binary mixtures of tall fescue (Schendonorus arundinaceus [Schreb.] Dumort, nom. Cons.) (TF) with alfalfa (ALF) (TF + ALF, Medicago sativa L.) and birdsfoot trefoil (BFT) (TF + BFT, Lotus corniculatus L.), to tall fescue with (TF + N) and without N fertilizer (TF-N). Pastures were established in Lewiston, Utah, USA, and rotationally stocked by Angus beef steers for 112 days in 2012 and 2013. Herbage samples were collected prior to each 7-day stocking period and analyzed for mass and nutritive value. Steers were weighed each 28-day rotation cycle and cumulative average daily gain (ADG) calculated. Steer performance differed (p = .05) among most treatments with ADG greatest for TF + BFT (0.73 kg/day) and TF + ALF (0.67 kg/day), followed by TF + N (0.61 kg/day) and the least for TF-N (0.40 kg/day). Multivariate analysis indicated that small inherent differences in herbage energy, fiber and digestibility were highly correlated with steer growth. Other variables associated with ADG included legume proportion and herbage mass. Fescue-legume mixtures had 29 to 33% legume proportion, and more (p = .05) favorable energy and fiber concentrations than grass monocultures. Whereas herbage mass was greatest (p = .05) for TF + N (2,956 kg/ha), followed closely by TF + BFT (2,741 kg/ha) and TF + ALF (2,623 kg/ha). Overall, TF + BFT and TF + ALF pastures had less herbage, but better nutritive value and increased steer growth performance compared to TF + N, without the added cost of fertilizer.

High necrotic enteritis (NE) incidence and mortality rates in poultry can be caused by Clostridium perfringens (CP) coinfected with Eimeria spp., a causative agent of coccidiosis. Banning of prophylactic use of antibiotics in feed has been accompanied by increased NE outbreaks, resulting in economically devastating losses to the broiler industry. To determine alternatives for controlling NE, we isolated CP-specific bacteriophages (BP), characterized their properties, evaluated their inhibitory effects on pathogenic CP, selected a highly effective phage (φCJ22), and used φCJ22 as a dietary supplement in experimental NE-afflicted broiler chickens. Male broilers (n = 780) were randomly assigned to 60 pens (n = 13 broilers/pen) and into 5 groups [CP-uninfected negative control (NC), basal diet (BD) without CP and BP; CP-infected positive control (PC), BD + CP; and 3 BP groups receiving low- (LP; BD + CP+105 BP), medium- (MP; BD + CP+106 BP), and high-phage (HP; BD + CP+107 BP plaque-forming units/kg) concentrations]. The results showed that MP and HP groups presented an antimicrobial activity toward clinical CP isolate strains, and the groups decreased NE lesions and mortality rates without changes in chicken performance at the end of the experimental period. After CP-challenge body weight gain and feed efficiency were significantly lower in phage-fed groups than that in the PC group (P < 0.05), and NE-associated mortality was the lowest in the HP group (P < 0.001). Moreover, histopathology revealed lesser gastrointestinal mucosal damage in the NC and BP-treated (LP, MP, and HP) groups than that in the PC group, and MP and HP significantly lowered viable CP number in the cecum content by up to 1.24log10 relative to only CP-infected PC group (P < 0.05). These findings suggest that addition of φCJ22 to chicken feed might effectively ameliorate NE, which is accompanied by reduced CP strains in the gut and compensate the performance of NE-afflicted broilers.

For lean meat production in swine industry, some synthetic compounds have been applied as feed additives in finishing swine feeds. However, this practice raises public concern due to potential imbalance of innate hormones coupled with a possible residue of the synthetic compounds in meat products after ingestion. In this study, we investigated natural phytogenic compounds (NPC) from fruit peel consisted of mainly ursolic and maslinic acids to enhance muscle development but decrease fat deposition in swine. To test effects on cell line, NPC was treated in the porcine primary cell line obtained from muscle tissue of piglets at approximately 6.5 kg of body weight and NIH-3T3-L1 cell line from mouse adipose tissue cell (Experiment 1). In the porcine primary cell line, immunofluorescence measurement indicated that myogenin expression increased at 20%, while the genes responsible for muscle development increased RNA abundance in MyoD at 20%, Mrf4 at 31%, PAX3 at 19%, and PAX7 at 16% observed in myotube development from myoblast when treated with NPC (P < 0.05). In NIH-3T3L1 cell, NPC treatment increased suppression on lipid accumulation at 40% (P < 0.05). In a follow-up in vivo investigation in a completely randomized design (Experiment 2), the same NPC tested in Experiment 1 at 0.53% dry matter (DM) did not affect intake of DM, whereas it tended to increase average daily gain compared with control without NPC supplementation (1.03 vs. 0.99; P = 0.08), leading to a tendency to increase gain-to-feed ratio (0.338 vs. 0.316; P = 0.12). While NPC supplementation did not influence hot carcass weight, animals treated with NPC decreased backfat thickness compared to those fed control (23.5 vs. 24.8 mm; P < 0.01). In summary, NPC from fruit improved growth performance and carcass trait due to their impacts on muscle cell development and fat deposition.

Abstract Background General antiviral agents such as oseltamivir are associated with certain adverse effects and the emergence of resistance. This study investigated the phytochemical properties, antiviral activities, and safety of three herbs used in traditional Korean medicine. Methods Extracts of three medicinal herbs (Brassica juncea, Forsythia suspensa, and Inula britannica) were prepared using ethanol or water. The total phenolic, flavonoid, and saponin content, condensed tannin content, and reducing sugar content of the herb extracts were determined via phytochemical screening. Tandem mass analysis was performed using an ultra-performance liquid chromatography (UPLC)-electrospray ionization (ESI)-Q/Orbitrap instrument. Virus titrations were determined via tissue culture infective dose (TCID50) and cytotoxicity assays. Hemolysis and hepatotoxicity were measured to determine safety. Results Among the three medicinal herbs, F. suspensa showed the highest concentration of phenolic compounds, flavonoids, and saponins. The number of phytochemical compounds detected via tandem mass analysis of B. juncea, F. suspensa, and I. britannica was 5 (including sinigrin, m/z [M-H] = 358.02), 14 (including forsythoside A, m/z [M-H] = 623.19), and 18 (including chlorogenic acid, m/z [M-H] = 353.20), respectively. The antiviral effects of the B. juncea extracts (ethanol and water) and I. britannica extract (ethanol) were further investigated. The ethanol extract of B. juncea showed a 3 Log TCID50/25 μL virus titration reduction and the water extract showed a selectivity index of 13.668 against infected influenza H1N1 virus A/NWS/33. The B. juncea extracts did not show hemolysis activities and hepatotoxicity (< 20%). The ethanol extract of I. britannica showed the most effective virus titration decrease, whereas its hemolytic and hepatotoxicity values were the most significantly different compared to the control. Despite the high concentration of phytochemicals detected in F. suspensa, the extract showed approximately 1 Log TCID50/25 μL at the highest concentration. Conclusion B. juncea may show antiviral effects against H1N1 in a host. In addition, B. juncea may also show decreased disadvantages compared to other antiviral agents.

Lonicera japonica (LJ; honeysuckle) is used in traditional folk medicine in Korea and is a rich source of ascorbic acid and phenolic components that are reported to have antioxidant and antibiotic properties. We performed an in vitro experiment to assess the effects of LJ extracts (LJE) on ruminal fermentation. Timothy hay (0.3 g dry matter [DM]) was incubated with buffer, ruminal fluid, and 0%, 3%, 5%, 7%, and 9% LJE. Batch culture fermentation was conducted separately for 12, 24, and 48 hr to determine gas production (GP), ruminal fermentation characteristics, and microbial population characteristics. The effects on GP were generally similar to those on DM degradability, with a linear decrease observed at 9% extract at 24 hr. NH3 -N showed a linear increase with increasing extract concentrations at 12 hr, whereas a decrease was observed at 24 hr. Extract supplementation decreased methane (CH4 ) production at 12, 24, and 48 hr. In addition, the abundance of fibrolytic bacteria and ciliate-associated methanogen was reduced at all concentrations of extracts. These results indicate that LJE have the potential to serve as a ruminal fermentation modifier to suppress CH4 production with minimal effects on nutrient digestion in the rumen.

In this study, co-infection of white spot syndrome virus (WSSV) and AHPND-Vibrio parahaemolyticus (Vp(AHPND)), was studied in juvenile Penaeus vannamei under the laboratory condition. Experimental shrimp were divided into 4 groups (group 1: WSSV only; group 2: Vp(AHPND) only; group 3: WSSV + Vp(AHPND); group 4: not challenged), and for the co-infection, shrimp were challenged with dual pathogens (initially exposed to WSSV and subsequently exposed to Vp(AHPND)). During the experiments, we observed an increased mortality rate in the co-infection group (group 3, N = 7/20 shrimp at day 2 post-infection [p.i.], and N = 15/20 shrimp at day 3 p.i.) than other groups (N = 1/20 shrimp at day 3 p.i., and N = 10/20 shrimp at day 4 p.i. in group 2; no mortalities in group 1 and 4). Also, we found obvious differences in WSSV infection among groups; strong WSSV-positives were detected by immunohistochemistry examination (brown coloration) and qPCR assay (the copy numbers between 2.0 x 10(3)-3.7 x 10(4)) in the co-infection group on the termination day, but these were not detected in the other groups. In addition, there were prominent differences in the histopathology of the hepatopancreas of surviving shrimp among groups; shrimp of the co-infection group showed typical histopathology signs of AHPND in the hepatopancreas, with sloughing of hepatopancreatic tubule epithelial cells (G4) on the termination day, but the hepatopancreas in shrimp of the other groups appeared normal. This study demonstrated that shrimp initially exposed to WSSV allows Vp(AHPND) to cause faster and higher mortality than shrimp infected with Vp(AHPND) alone, and also leads to severe WSSV infection, indicating that WSSV is a risk factor for Vp(AHPND) in ponds. Therefore, shrimp farmers should pay attention to the management of co-infection in the pond.

Background Rheumatoid arthritis (RA) is a chronic and systemic inflammatory disease. The incidence of cardiovascular (CV) disease is increased in patients with RA, compared with the general population, which is related to the fact that atherosclerosis has an inflammatory etiology. Several studies revealed that RA is associated with systemic bone loss, and long-term glucocorticoid therapy is also known to affect CV events as well as bone health such as osteoporosis. Especially in postmenopausal women, the prevalence of osteoporosis and its complications are important medical issues. Objectives In the present study, we investigated the bone mineral density (BMD) for the carotid plaque formation in RA patients in the Kyungpook National University Hospital Atherosclerosis Risk in Rheumatoid Arthritis (KARRA) cohort study. Methods After a baseline evaluation for KARRA enrollment, RA patients were prospectively followed up for 5 years or until deaths. We analysed the demographic findings, conventional CV risk factors and RA disease activity. Carotid ultrasound at baseline and year 5 was performed to evaluation of the intima-medial thickness (IMT) and presence and progression of carotid plaque. A total 323 patients (272 female) with RA, who performed dual-photon x-ray absorptiometry and carotid ultrasound, were included. We assessed disease activity of RA, risk factors for atherosclerosis including hypertension, diabetes mellitus and dyslipidemia, presence of carotid plaque, BMD and cumulative glucocorticoid doses. Results A total of 417 RA patients were included in the baseline KARRA cohort, and 327 patients with RA were followed for the 5 year period. Of the 417 baseline RA patients, 212 patients had no carotid plaque. At year 5, new carotid plaque formation was found in 91 of 214 patients who underwent BMD examination. The BMD in the l-spine, femur, and radius was significantly lower in patients with carotid plaques (n=154), compared to patients without plaques (n=172) (1.016 g/cm2±0.22 vs. 1.066±0.18, p=0.013 for l-spine; 0.817±0.15 vs. 0.865±0.14, p<0.001 for femur; 0.542±0.14 vs. 0.605±0.13, p<0.001 for radius. In postmenopausal patients, the BMD was significantly lower in carotid plaque group (n=93) than non-plaque group (n=109) (0.962±0.171 vs. 1.056±0.174, p<0.001 for L spine; 0.780±0.14 vs. 0.857±0.12, p<0.001 for femur; 0.502±0.110 vs. 0.593±0.112, p<0.001 for radius). The cumulative steroid dose was confirmed in postmenopausal female patients, and the glucocorticoid dose was correlated with new carotid plaque formation. (r=0.334, p=0.04). Multivariate logistic regression analysis revealed that but radius BMD (p=0.04) was independent risk factors for new carotid plaque formation during the 5 year followed period after correlation with cumulative glucocorticoid dose, but l-spine (p=0.06) and femur BMD (p=0.07) were not statistically significant. Conclusions This study shows formation of new plaques after long-term follow-up depends on the juxta-articular bone health in postmenopausal RA patients. Disclosure of Interest None declared