Genome Sequence and Characterization of Acinetobacter Phage DMU1.

Background: There has been a recent resurgence of research on the characterization of Acinetobacter phage for therapeutic use due to the morbidity and mortality associated with treatment failures in cases of multidrug-resistant Acinetobacter baumannii infections. Materials and Methods: A bacteriophage isolated from activated sludge that targets A. baumannii ATCC19606 was characterized by electron microscopy, genome sequencing, comparative genomics, and a host range analysis. Results: The morphology of Acinetobacter phage DMU1 resembles phages in Siphoviridae. Comparative genomic and phylogenetic analyses reveal that DMU1 is a siphophage and is most closely related to Acinetobacter phage SH-Ab 15497. Out of the strains tested, DMU1 was found to only infect A. baumannii strains ATCC19606 and ATCC17978. Conclusion: Acinetobacter phage DMU1 belongs to the Siphoviridae family and is most closely related to Acinetobacter phage SH-Ab 15497. Small-scale host-range analysis of DMU1 indicates a host range that is likely limited to specific A. baumannii strains.

Influence of maternal nutrient restriction and rumen-protected arginine supplementation on post-ruminal digestive enzyme activity of lamb offspring.

To determine the influence of maternal nutrient restriction and rumen-protected arginine supplementation on post-ruminal digestive enzyme activity in lambs, 31 multiparous, Rambouillet ewes were allocated to one of three dietary treatments at 54 d of gestation. Dietary treatments were 100% of nutrient requirements (control, CON; n = 11), 60% of control (restricted, RES; n = 10), or RES plus 180 mg rumen-protected arginine•kg BW-1•d-1 (RES-ARG; n = 10). Immediately after parturition, lambs were removed from dams and reared independently. Milk-replacer and alfalfa hay + creep feed were offered for ad libitum intake. At day 54 of age, lambs were slaughtered and the pancreas and small intestine were collected. Pancreatic (α-amylase and trypsin) and jejunal (maltase, glucoamylase, sucrase, isomaltase, and lactase) digestive enzyme activities were assayed. Data were analyzed using the GLM procedure of SAS for effects of treatment. Contrast statements were used to determine differences between means for effects of restriction (CON vs. RES and RES-ARG) and rumen-protected arginine supplementation (RES vs. RES-ARG). There was no influence (P ≥ 0.15) of maternal nutrient restriction or rumen-protected arginine supplementation on pancreatic or jejunal protein concentrations. No treatment effects were observed (P ≥ 0.12) for enzymes involved in starch digestion including pancreatic α-amylase and jejunal maltase, glucoamylase, and isomaltase. Sucrase activity was undetected in the jejunum of lambs across all treatments. Maternal nutrient restriction tended to increase (P = 0.08) pancreatic trypsin activity per gram protein in lambs. Lactase activity per gram protein in the jejunum of lambs tended to decrease (P = 0.09) with maternal nutrient restriction. Rumen-protected arginine supplementation to gestating ewes did not influence (P ≥ 0.19) digestive enzyme activities of lamb offspring. These data suggest that maternal nutrient restriction and rumen-protected arginine supplementation have minimal effects on digestive enzyme activity in offspring.

Loss of the nutrient receptor Tas1R3 reduces atherosclerotic plaque accumulation and hepatic steatosis in ApoE-/- mice.

The taste receptor type I (Tas1R) family consists of three G protein-coupled receptors (T1R1, T1R2, and T1R3) that form heterodimers recognizing sweet compounds (T1R2/T1R3) or amino acids (T1R1/T1R3). These receptors are nutrient sensors that facilitate appropriate physiological responses with nutrient availability. However, their contribution to the development of pathologies associated with overnutrition (e.g., atherosclerosis) is unclear. The aim of the present study was to determine if T1R3 deletion would reduce atherosclerotic plaque development in mice. We generated atherosclerotic mice with whole-body deletion of T1R3 by crossing T1R3-/- mice with ApoE-/- mice. T1R3+/+ ApoE-/- and T1R3-/- ApoE-/- mice were maintained on an atherogenic high-fat diet for 8 weeks. Weight gain and food consumption were measured during the 8-week diet. Atherosclerotic lesion development and size were assessed by en face analysis of intact aortas and microscopic analysis of aortic roots. Our results indicate that T1R3 deletion in male and female ApoE-/- mice reduces aortic atherosclerotic plaque accumulation. Hepatic triglyceride accumulation, which was measured by quantification of oil red O staining, was also reduced in T1R3-/- mice. While the ablation of T1R3 reduced the final body weight of both males and females by approximately 12%, serum lipids, insulin, and glucose were either unchanged or slightly reduced. Immunoblot analysis of the phosphorylation of p70S6K, an effector of mTORC1, suggests T1R3 ablation reduces mTORC1 activity by approximately 50% in the male livers. Collectively, these findings suggest that the whole-body deletion of T1R3 reduces atherosclerosis and hepatic steatosis in a manner largely independent of the measured effects on whole-body glucose and lipid homeostasis.

In vitro evaluation of nano zinc oxide (nZnO) on mitigation of gaseous emissions.

BACKGROUND: Enteric methane (CH4) accounts for about 70% of total CH4 emissions from the ruminant animals. Researchers are exploring ways to mitigate enteric CH4 emissions from ruminants. Recently, nano zinc oxide (nZnO) has shown potential in reducing CH4 and hydrogen sulfide (H2S) production from the liquid manure under anaerobic storage conditions. Four different levels of nZnO and two types of feed were mixed with rumen fluid to investigate the efficacy of nZnO in mitigating gaseous production. METHODS: All experiments with four replicates were conducted in batches in 250 mL glass bottles paired with the ANKOMRF wireless gas production monitoring system. Gas production was monitored continuously for 72 h at a constant temperature of 39 ± 1 °C in a water bath. Headspace gas samples were collected using gas-tight syringes from the Tedlar bags connected to the glass bottles and analyzed for greenhouse gases (CH4 and carbon dioxide-CO2) and H2S concentrations. CH4 and CO2 gas concentrations were analyzed using an SRI-8610 Gas Chromatograph and H2S concentrations were measured using a Jerome 631X meter. At the same time, substrate (i.e. mixed rumen fluid+ NP treatment+ feed composite) samples were collected from the glass bottles at the beginning and at the end of an experiment for bacterial counts, and volatile fatty acids (VFAs) analysis. RESULTS: Compared to the control treatment the H2S and GHGs concentration reduction after 72 h of the tested nZnO levels varied between 4.89 to 53.65%. Additionally, 0.47 to 22.21% microbial population reduction was observed from the applied nZnO treatments. Application of nZnO at a rate of 1000 µg g- 1 have exhibited the highest amount of concentration reductions for all three gases and microbial population. CONCLUSION: Results suggest that both 500 and 1000 µg g- 1 nZnO application levels have the potential to reduce GHG and H2S concentrations.

Influence of feeding direct-fed microbial supplementation on growth performance and feeding behavior in naturally fed and conventionally fed finishing cattle with different dietary adaptation periods.

To determine the effects of finishing system (conventional vs. natural), dietary adaptation length (14 vs. 28 d), and direct-fed microbial (DFM) supplementation (no DFM vs. DFM) on growth performance and feeding behavior, 120 yearling steers (390 ± 2.8 kg) were used in a completely randomized design with a 2 × 2 × 2 factorial arrangement of treatments. Feed intake was monitored using the Insentec feeding system. Blood samples were collected every 28 d. After 140 d on feed, steers were slaughtered and carcass characteristics collected. Conventionally fed steers had greater (P ≤ 0.001) final BW, carcass weight, and dressing %. Dry matter intake was not influenced (P ≥ 0.31) by treatment. Length of dietary adaptation period did not influence (P ≥ 016) final BW, ADG, and G:F. There was a feeding system × DFM interaction (P ≤ 0.02) for ADG and G:F with conventionally fed steers fed DFM having the greatest (P ≤ 0.05) and naturally fed steers supplemented with DFM having the least (P ≤ 0.05) ADG and G:F. Number of visits to the feed bunk and number of meals per day did not differ (P > 0.05) among treatments. Time eating per visit and per meal was greater (P = 0.05) in steers supplemented with DFM than in steers not supplemented with DFM. On day 56 and 140, plasma glucose concentration was greater (P ≤ 0.03) in steers adapted in 14 d than in steers adapted in 28 d. On day 84, plasma glucose concentration was greater (P = 0.02) in naturally fed compared to conventionally fed steers. On day 112, there was a dietary adaptation period × DFM interaction (P = 0.004) for plasma glucose concentration with glucose concentration greatest (P ≤ 0.05) in steers adapted in 14 d supplemented with DFM and in steers adapted in 28 d not supplemented with DFM, least for steers adapted in 28 d supplemented with DFM, with steers adapted in 14 d not supplemented with DFM intermediate (P ≤ 0.05). On day 112 and 140, plasma urea N concentration was greater (P ≤ 0.05) in steers adapted in 28 d than in steers adapted in 14 d. These data indicate that conventionally fed steers generally had improved growth performance compared to naturally fed steers. Length of dietary adaptation and DFM supplementation had minimal effects on growth performance but did interact with feeding system to influence feeding behavior and blood metabolite concentrations.