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The objective of this study was to evaluate the antibacterial activities of extract derived from moringa leaves. In particular, the effect of moringa extract (Mor) on adhesion and invasion of Escherichia coli O55, Enterococcus faecalis, Staphylococcus simulans, and Serratia liquefaciens was evaluated in bovine mammary epithelial cells (MAC-T). Broth microdilution method, minimum inhibitory concentration and minimum bactericidal concentration assays, adhesion and invasion assays, and real-time PCR were performed. The minimum inhibitory concentration and minimum bactericidal concentration of Mor ranged from 12.5 to 50 mg/mL on 18 out of 27 tested isolates. Treatment of E. coli O55 with Mor (100 and 200 μg/mL) inhibited the adhesion and invasion on MAC-T cells via downregulation of adhesion factors (i.e., papC, f17c-A, and eaeA). Also, when MAC-T cells were pretreated with Mor (200 μg/mL, 12 h) and incubated with E. coli O55, Enterococcus faecalis, Staphylococcus simulans, or Serratia liquefaciens, both E. coli O55 and Enterococcus faecalis showed a significant decrease in adhesion and invasion. Staphylococcus simulans exhibited decreased adhesion and increased invasion. Serratia liquefaciens showed increased adhesion and decreased invasion. In addition, Mor increased mRNA expression of antioxidant enzymes (e.g., heme oxygenase-1, NAD(P)Hquinone oxidoreductase-1, and thioredoxin reductase 1) in MAC-T cells. In conclusion, 12.5 to 50 mg/mL of Mor exhibited antibacterial activity against 18 out of 27 tested isolates. Also, pretreatment of 200 μg/mL of Mor to MAC-T cells modulated adhesion and invasion of E. coli O55 and other mastitis-associated pathogens. Furthermore, Mor increased antioxidant capacities in MAC-T cells, but further in vivo studies are needed.Escherichia coli O157H7 is a common harmful foodborne pathogen that can cause severe diseases at low infectious doses. Traditional lateral flow immunoassay (LFIA) for the rapid screening of E. coli O157H7 in food suffers from low sensitivity due to its dependence on 20- to 40-nm gold nanoparticles (AuNP) with insufficient brightness as labels. To address this issue, we reported for the first time the successful synthesis of gold superparticles (GSP) by encapsulating numerous small AuNP into a polymer nanobead using an evaporation-induced self-assembly method. Results indicated that the resultant GSP exhibited remarkably enhanced absorbance compared with the most widely used 40 nm AuNP in LFIA. In addition, the absorbance of GSP could be easily tuned by varying GSP sizes. Under optimized conditions, we achieved a rapid and sensitive determination of E. coli O157H7 in milk with a detection limit of 5.95 × 102 cfu/mL when using the GSP with a size of 342 nm as LFIA signal reporters, exhibiting improvement of approximately 32-fold relative to the conventional 40 nm AuNP-LFIA method. We further demonstrated the selectivity, accuracy, reliability, and practicality of the proposed GSP-LFIA strip. In summary, this work offers a new strategy for improving LFIA sensitivity using assembled GSP as markers and demonstrates huge potential in rapidly and sensitively detecting foodborne pathogens.Dairy cows are often diagnosed with fever without showing clinical symptoms of disease. The aim of this study was to investigate changes in feeding, social, and lying behaviors of cows with fever but without clinical disease, as compared with healthy cows. After parturition, dairy cows of mixed parities were housed in a dynamic group of 20. In the freestall pen, cows had access to 12 electronic feed bins, 2 electronic water bins, and 24 lying stalls. Feeding and social behaviors were recorded using the electronic feed bins, and lying behaviors were measured using electronic data loggers attached to the cow. Rectal body temperature was assessed on a daily basis, and fever defined as a body temperature >39.5°C. All cows were examined for metritis every third day after calving, and all other diseases (e.g., mastitis, ketosis) were diagnosed as per farm protocol. Cows with multiple days of fever (n = 8) and cows with 1 d of fever (n = 18) that were not diagnosed with a clinical disease were compared with a matched sample of healthy cows (i.e., cows that were not clinically ill and never had a fever recorded) of the same parity (categorized as primiparous vs. multiparous). Feeding, social, and lying behaviors were compared for the first 2 d of fever in cows with multiple days of fever, and the day of fever in cows with 1 d of fever. Cows of both fever groups spent less time feeding compared with controls (135 vs. 181 ± 7.6 min/d for multiple fever days, and 158 vs. 185 ± 9.7 min/d for 1 d of fever). Cows with 1 d of fever ate at a faster rate (109 vs. 91 ± 5 g/min) and had a lower number of replacements at the feed bunk (actor replacements 9.7 vs. 14.6 ± 1.7 no./d; reactor replacements 11.1 vs. 15.9 ± 1.6 no./d) compared with healthy controls. Overall, cows with fever showed behavioral changes such as decreased feeding time that are consistent with sickness responses described in other species.Records of methane emissions from 1,501 cows on 14 commercial farms in 4 regions of Spain were collected from May 2018 to June 2019. Methane concentrations (MeC) were measured using a nondispersive infrared methane detector installed within the feed bin of the automatic milking system during 14- to 21-d periods. Rumination time (RT; min/d) was collected using collars with a tag that registered time (minutes) spent eating and ruminating. The means of MeC and methane production (MeP) were 1,254.28 ppm and 182.49 g/d, respectively; mean RT was 473.38 min/d. Variance components for MeC, MeP, and RT were estimated with REML using pedigree and genomic information in a single-step model. Heritabilities for MeC and MeP were 0.11 and 0.12, respectively. Rumination time showed a slightly larger heritability estimate (0.17). The genetic correlation between MeP and MeC was high (>0.95), suggesting that selection on either trait would lead to a positive correlated response on the other. Negative correlations were estimatetions were also estimated for stature with MeC and MeP (0.30 ± 0.04 and 0.43 ± 0.04, respectively). Other type traits (chest width, udder depth, angularity, and capacity) were positively correlated with methane traits, possibly because of higher milk yield and higher feed intake from these animals. Rumination time showed positive EBV correlations with production traits and type traits, and negative correlations with somatic cell count and body condition score. Based on the genetic correlations and heritabilities estimated in this study, methane is measurable and heritable, and estimates of genetic correlations suggest no strong opposition to current breeding objectives in Spanish Holsteins.Postpasteurization contamination (PPC) with gram-negative bacteria adversely affects the quality and shelf-life of milk through the development of flavor, odor, texture, and visual defects. Through evaluation of milk quality at 4 large fluid milk processing facilities in the northeast United States, we examined the efficacy of 3 strategies designed to reduce the occurrence of PPC in fluid milk (1) employee training (focusing on good manufacturing practices) alone and (2) with concurrent implementation of modified clean-in-place chemistry and (3) preventive maintenance (PM) focused on replacement of wearable rubber components. Despite increases in employee knowledge and self-reported behavior change, microbiological evaluation of fluid milk before and after interventions indicated that neither training alone nor training combined with modified clean-in-place interventions significantly decreased PPC. Furthermore, characterization of gram-negative bacterial isolates from milk suggested that specific bacterial t improvements in PM and sanitary equipment design may not always lead to reduced PPC. Our data also indicate that although short-term evaluations, such as pre- and post-tests for employee training, may suggest improvements after corrective and preventive actions, extensive microbial testing, ideally in combination with isolate characterization, may be necessary to evaluate return on investment of different interventions.In this study, heated whey protein isolate and pectin complexes (HCPX) formed at pH > isoelectric point (pI) were used to stabilize oil-in-water emulsions containing 5% oil and 1.5% (wt%) protein at pH 5.5. this website The effects of pectin concentration and heating temperature on emulsification and emulsion stabilization properties were determined. The HCPX were produced by heating mixed 3% (wt) whey protein isolate and pectin (0.1 or 0.3 wt%) at pH 6.2 and 75 or 85°C for 15 min. Aggregate sizes significantly increased with increasing heating temperature but decreased with the addition of pectin. The HCPX became more negatively charged with increasing pectin concentration; however, the effect of heating temperature was significant only at 0.1% pectin. Unheated complexes and HCPX successfully adsorbed at the oil-in-water interface and improved the emulsification properties as shown by higher negative charge and smaller droplet sizes. Despite the presence of pectin, rheological properties of the emulsions were not significantly different. All complexes showed increased emulsion stability; however, HCPX made at 85°C formed emulsions that were the most stable against creaming and heating.A continuous-culture fermentor study was conducted to assess nutrient digestibilities, volatile fatty acid (VFA) concentrations, microbial protein synthesis, bacterial nitrogen (N) efficiency, and enteric methane (CH4) production of four 5050 grass-legume diets, randomly assigned in a 4 × 4 Latin square design. Four legumes with different concentrations of condensed tannins (CT) were tested alfalfa [ALF; Medicago sativa L., non-CT legume]; birdsfoot trefoil [BFT; Lotus corniculatus L., low-CT legume]; crown vetch [CV; Securigera varia (L.) Lassen, moderate-CT legume]; and sericea lespedeza [SL; Lespedeza cuneata (Dum. Cours.) G. Don, high-CT legume]. Orchardgrass (Dactylis glomerata L.) was the common forage used in all diets. Four fermentors were evaluated over four 10-d periods by feeding 82 g of dry matter (DM)/d in 4 equal feedings. link2 Methane output was recorded every 10 min. link3 Effluent samples were collected during the last 3 d of the experiment, composited by fermentor and period, and analyzed for pH and VFA, as well as DM, organic matter, crude protein, neutral detergent fiber, and acid detergent fiber for determination of apparent and true nutrient digestibilities. Microbial protein synthesis and bacterial efficiency were estimated by analysis of N flows and purines. The CT concentrations were 3, 21, 38 and 76 g/kg of DM for ALF, BFT, CV, and SL diets, respectively. The SL diet had decreased fiber digestibilities and total VFA concentrations compared with the other diets. This resulted in the least total CH4 production in the SL diet. Bacterial N efficiency per kilogram of organic matter truly digested was lower in the SL diet than in the BFT and CV diets. The lowest CH4 production per unit of digestible nutrients was also found in the SL diet. Further work should be conducted to find optimal diets (by testing other legumes, rations, and sources of CT) for reducing CH4 emissions without negatively affecting ruminal digestion to maintain or improve productivity.
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