Determining Mancozeb Deposition Benchmark Values on Apple Leaves for the Management of Venturia inaequalis.

Apple scab, caused by Venturia inaequalis, is the most common fruit and foliar disease in commercial apple production worldwide. Early in the production season, preventative contact fungicide sprays are essential for protecting highly susceptible continuously unfolding and expanding young leaves. In South Africa, mancozeb is a key contact fungicide used for controlling apple scab early in the season. The current study developed deposition benchmarks indicative of the biological efficacy of mancozeb against apple scab, using a laboratory-based apple seedling model system. The model system employed a yellow fluorescent pigment that is known to be an effective tracer of mancozeb deposition. A concentration range of mancozeb (0.15 to 1 times the registered dosage) and fluorescent pigment concentrations was sprayed onto seedling leaves, which yielded various fluorescent particle coverage (FPC%) levels. Modeling of the FPC% values versus percent disease control yielded different benchmark values when disease quantification was conducted using two different methods. Thermal infrared imaging (TIRI) disease quantification resulted in a benchmark model where 0.40%, 0.79%, and 1.35 FPC% yielded 50, 75, and 90% apple scab control, respectively. These FPC% values were higher than the benchmarks (0.10, 0.20, and 0.34 FPC%, respectively) obtained with quantitative real-time PCR (qPCR) disease quantification. The qPCR benchmark model is recommended as a guideline for evaluating the efficacy of mancozeb sprays on leaves in apple orchards since the TIRI benchmark model underestimated disease control. The TIRI benchmark model yielded 68% disease control at the lowest mancozeb dosage, yet no visible lesion developed at this dosage. Both benchmark models showed that mancozeb yielded high levels of disease control at very low concentrations; for the qPCR benchmark model the FPC% value of the FPC90 (90% control) corresponded to 0.15 times that of the registered mancozeb concentration in South Africa, i.e., 85% lower than the registered dosage.

Population-based analysis of sociodemographic predictors, health-related quality of life and health service use associated with obstructive sleep apnoea and insomnia in Australia.

Although there is growing recognition of the effects of living with sleep disorders and the important role of primary care in their identification and management, studies indicate that the detection of sleep apnoea (OSA) and insomnia may still be low. This large representative community-based study (n=2977 adults) used logistic regression models to examine predictors of self-reported OSA and current insomnia and linear regression models to examine the association of these sleep conditions with both mental and physical components of health-related quality of life (HRQoL) and health service use. Overall, 5.6% (95% confidence interval (CI) 4.6-6.7) and 6.8% (95% CI 5.7-7.9) of subjects self-reported OSA (using a single-item question) and current insomnia (using two single-item questions) respectively. Many sociodemographic and lifestyle predictors for OSA and insomnia acted in different directions or showed different magnitudes of association. Both disorders had a similar adverse relationship with physical HRQoL, whereas mental HRQoL was more impaired among those with insomnia. Frequent consultations with a doctor were associated with a lower physical HRQoL across these sleep conditions; however, lower mental HRQoL among those frequently visiting a doctor was observed only among individuals with insomnia. The adverse relationship between sleep disorders and physical and mental HRQoL was substantial and should not be underestimated.

Pilot cohort study of obstructive sleep apnoea in community-dwelling people with schizophrenia.

OBJECTIVES: We aimed to assess the incidence of obstructive sleep apnoea (OSA) in people with schizophrenia, to explore clinical associates with OSA and how well OSA screening tools perform in this population. METHODS: All patients registered in a community outpatient Clozapine clinic, between January 2014 and March 2016, were consecutively approached to participate. Participants were screened for OSA using at home multichannel polysomnography (PSG) and were diagnosed with OSA if the apnoea-hypopnoea index (AHI) was >10 events/hr. Univariate comparison of participants to determine whether AHI > 10 events/hr was associated with demographic factors, anthropometric measures and psychiatric symptoms and cognition was performed. The sensitivity, specificity, positive predictive value and negative predictive value of the commonly used sleep symptoms scales and OSA screening tools were also determined. RESULTS: Thirty participants were recruited, 24 men and 6 women. Mean age was 38.8 (range: 25-60), and mean body mass index (BMI) was 35.7 (range 19.9-62.1). The proportion of participants with OSA (AHI > 10 events/hr) was 40%, 18 (60%) had no OSA, 4 (13%) had mild OSA (AHI 10.1-20), zero participants had moderate OSA (AHI 20.1-30) and 8 (27%) had severe OSA (AHI > 30). Diagnosis of OSA was significantly associated with increased weight, BMI, neck circumference and systolic blood pressure. Diagnosis of OSA was not significantly associated with Positive and Negative Symptoms Scale, Montgomery Asperger's Depression Rating Scale, Personal and Social Performance scale or Brief Assessment of Cognition for Schizophrenia scores. All OSA screening tools demonstrated poor sensitivity and specificity for a diagnosis of OSA. CONCLUSION: OSA was highly prevalent in this cohort of people with schizophrenia and was associated with traditional anthropometric OSA risk factors.

Expression of a bacterial ice nucleation gene in a yeast Saccharomyces cerevisiae and its possible application in food freezing processes.

A 3.6 kb ice nucleation gene (ina) isolated from Erwinia herbicola was placed under control of the galactose-inducible promoter (GAL1) and introduced into Saccharomyces cerevisiae. Yeast transformants showed increased ice nucleation activity over untransformed controls. The freezing temperature of a small volume of water droplets containing yeast cells was increased from approximately -13 degrees C in the untransformed controls to -6 degrees C in ina-expressing (Ina(+)) transformants. Lower temperature growth of Ina(+) yeast at temperatures below 25 degrees C was required for the expression of ice nucleation activity. Shift of temperature to 5-20 degrees C could induce the ice nucleation activity of Ina(+) yeast when grown at 25 degrees C, and maximum ice nucleation activity was achieved after induction at 5 degrees C for approximately 12 h. The effects of Ina(+) yeast on freezing and texturization of several food materials was also demonstrated.

Control of enzymatic browning in potato (Solanum tuberosum L.) by sense and antisense RNA from tomato polyphenol oxidase.

Polyphenol oxidase (PPO) activity of Russet Burbank potato was inhibited by sense and antisense PPO RNAs expressed from a tomato PPO cDNA under the control of the 35S promoter from the cauliflower mosaic virus. Transgenic Russet Burbank potato plants from 37 different lines were grown in the field. PPO activity and the level of enzymatic browning were measured in the tubers harvested from the field. Of the tubers from 28 transgenic lines that were sampled, tubers from 5 lines exhibited reduced browning. The level of PPO activity correlated with the reduction in enzymatic browning in these lines. These results indicate that expression of tomato PPO RNA in sense or antisense orientation inhibits PPO activity and enzymatic browning in the major commercial potato cultivar. Expression of tomato PPO RNA in sense orientation led to the greatest decrease in PPO activity and enzymatic browning, possibly due to cosuppression. These results suggest that expression of closely related heterologous genes can be used to prevent enzymatic browning in a wide variety of food crops without the application of various food additives.

Supplemental betaine and peroxide-treated feather meal for finishing cattle.

These studies evaluated the effects of betaine, provided either as feed-grade betaine or as concentrated separator by-product (CSB; desugared beet molasses), on performance and carcass characteristics of finishing cattle. In Exp. 1, 175 steers (410 kg initial BW) were fed a finishing diet based on steam-flaked and dry-rolled corn, and treatments included 10.5 and 21 g/d feed-grade betaine and 250 and 500 g/d CSB (supplying 15.5 and 31 g/d of betaine, respectively). Steers fed feed-grade betaine had greater (linear and quadratic effects, P < 0.1) DMI than control steers, but ADG and gain efficiencies were not affected by treatment. Dressing percent and backfat thickness was greater (P < 0.1) for steers that received feed-grade betaine than for controls. Longissimus muscle area was lower (P < 0.1) for steers supplemented with either feed-grade betaine or CSB than for control steers. Yield grades were higher for cattle receiving feed-grade betaine (quadratic effect, P < 0.1) than for control steers. Marbling scores were not affected by supplemental betaine, but the percentage of carcasses grading USDA Select was lower (linear and quadratic effects, P < 0.1) for steers fed feed-grade betaine than for control steers, predominantly due to a greater percentage grading USDA Choice. In Exp. 2, 312 heifers (343 kg initial BW) were used in a finishing study to evaluate the effects of graded levels of feed-grade betaine and peroxide-treated feather meal on performance and carcass characteristics. Treatments included two finishing diets (containing peroxide-treated or untreated feather meal) and four levels (0, 4, 8, and 12 g/d) of feed-grade betaine arranged in a 2 x 4 factorial. No significant interactions occurred between treatment of feather meal and betaine. Treatment of feather meal with hydrogen peroxide (5% wt/wt) increased in situ protein degradability but did not alter DMI, ADG, gain efficiencies, or carcass characteristics of heifers when it replaced untreated feather meal in the diet. Top-dressing feed-grade betaine to the diets had no effect on DMI, ADG, and gain efficiencies. Marbling scores were greater (cubic effect, P < 0.05) for heifers fed diets top-dressed with 4 and 12 g/d of feed-grade betaine, but other carcass characteristics were not altered significantly. Overall, feed-grade betaine and CSB did not alter growth performance, but did have minor effects on carcass characteristics.

Wet corn gluten feed and alfalfa hay combinations in steam-flaked corn finishing cattle diets.

One finishing trial and one digestibility trial were used to evaluate wet corn gluten feed (WCGF) and alfalfa hay (AH) combinations in steam-flaked corn (SFC) finishing diets. In Exp. 1, 631 crossbred heifers (initial BW = 284 +/- 7.9 kg) were fed SFC-based diets containing combinations of WCGF (25, 35, or 45% of diet DM) and AH (2 or 6% of dietary DM) in a 2 x 3 factorial arrangement of treatments. No interactions existed between WCGF and AH for heifer performance. Increasing dietary WCGF linearly decreased gain efficiency (P < 0.01), dietary NEg concentration (P < 0.05), and 12th-rib fat thickness (P = 0.10). Cattle fed 35% WCGF had the lowest occurrence of abscessed livers, resulting in a quadratic response (P < 0.05) as dietary WCGF increased. In Exp. 2, 12 ruminally cannulated Jersey steers (585 kg) were fed SFC-based diets containing combinations of WCGF (25 or 45% of diet DM) and AH (0, 2, or 6% of diet DM) in an incomplete Latin square design with a 2 x 3 factorial arrangement of treatments. Starch intake was lower (P < 0.05), but NDF intake was greater (P < 0.05) as AH and WCGF increased in the diet. Ruminal pH was increased by AH (linear, P < 0.05) and tended (P < 0.07) to increase with WCGF. Feeding 2% AH led to the greatest ruminal NH3 but the lowest total VFA and propionate (quadratic, P < 0.05). Addition of AH to diets containing 25% WCGF increased acetate to a greater extent than addition to diets containing 45% WCGF (AH x WCGF interaction, P < 0.05). Feeding 45% WCGF tended to increase passage rate (P = 0.17) and decrease (P < 0.05) total tract OM digestibility but increase (P < 0.05) in situ degradation of DM from AH and WCGF. Interactions between AH and WCGF existed (P < 0.05) for ruminal fluid volume (quadratic effect of AH x WCGF level), in situ SFC degradation (linear effect of AH x WCGF level), and in situ rate of WCGF DM disappearance (quadratic effect of AH x WCGF level). We conclude that AH levels may be decreased when WCGF is added to SFC diets as 25% or more of the dietary DM.

Effect of Maillard reaction products on ruminal and fecal acid-resistant E. coli, total coliforms, VFA profiles, and pH in steers.

Six ruminally cannulated Angus-cross steers (362 kg) were used in a replicated 3 x 3 Latin square design to determine effects of supplementing Maillard reaction products (MRP) on acid-resistant E. coli and coliform populations. Steers were fed roughage-based diets supplemented (DM basis) with either 10% soybean meal (SBM), 10% nonenzymatically browned SBM (NESBM), or 10% SBM top-dressed with 45 g of a lysine-dextrose Maillard reaction product (LD-MRP). Equal weights of dextrose, lysine hydrochloride, and deionized water were refluxed to produce the LD-MRP. The NESBM was manufactured by treating SBM with invertase enzyme, followed by heating to induce nonenzymatic browning. Steers were allowed slightly less than ad libitum access to diets fed twice daily and were adapted to their respective treatments within 10 d. On d 11, ruminal and fecal samples were collected at 0, 2, 4, 6, 8, and 12 h after feeding from each of the steers and transported to the laboratory for microbial analysis. Ruminal samples and feces were analyzed for pH and VFA, and both ruminal fluid and feces were tested for acid-resistant E. coli and total coliforms by incubating samples in tryptic soy broth adjusted to pH 2, 4, and 7. Ruminal pH and total VFA concentrations did not differ among treatments. The molar proportion of ruminal acetate was higher (P < 0.05) for steers receiving NESBM than for steers receiving SBM and LD-MRP. At pH 4, steers that received NESBM had lower (P < 0.05) ruminal populations of E. coli and total coliforms than steers that received SBM. No differences were observed for ruminal E. coli and total coliforms at pH 2 and 7. Fecal pH was lower (P < 0.05) for steers fed NESBM than for steers fed SBM or LD-MRP. Molar proportions of fecal acetate were lower (P < 0.05) and proportions of butyrate and isovalerate were higher (P < 0.05) for steers fed NESBM compared with steers fed SBM. Fecal E. coli at pH 4 was lower (P < 0.05) for steers fed NESBM than for steers fed LD-MRP. Fecal E. coli and total coliforms at pH 2 and 7 did not differ among treatments. Dietary MRP had limited effectiveness at decreasing acid-resistant coliforms in the rumen and feces of cattle. Acid resistance in coliforms may depend on protein availability.

Combinations of alfalfa hay and wet corn gluten feed in limit-fed growing diets for beef cattle.

Two experiments were conducted to evaluate the effects of alfalfa hay (AH) and wet corn gluten feed (WCGF) combinations on ADG and gain efficiency of cattle limit-fed growing diets. In Exp. 1, crossbred beef steers (n = 220; initial BW = 262 kg) were limit-fed diets consisting of steam-flaked corn and 40% WCGF (DM basis) with 0, 10, or 20% ground AH (0AH, 10AH, and 20AH, respectively). A fourth diet containing 20% ground AH and steam-flaked corn served as a control. All diets were fed once daily at 1.8% of BW (DM basis). Growing period ADG, gain efficiency, and dietary NE calculated from performance data decreased linearly (P < 0.01) with addition of AH to diets containing WCGF. Rate of DMI increased linearly (P < 0.05) with AH addition to diets containing WCGF. Following the growing period, steers were finished on a common diet offered ad libitum. Gain efficiencies during the finishing period were higher (P < 0.05) for steers fed the 20AH diet than for steers fed the control diet. In Exp. 2, crossbred beef heifers (n = 339; initial BW = 277 kg) were limit-fed diets containing steam-flaked corn with 10, 20, or 30% ground AH and 0, 40, or 68% WCGF in a 3 x 3 factorial arrangement, fed once daily at 1.6% of BW (DM basis). An AH x WCGF interaction occurred (P < 0.05) for growing period ADG and gain efficiency. Increasing AH or WCGF decreased cattle ADG, gain efficiency, and dietary NE with the exception of heifers fed 30AH/40WCGF, which had ADG that did not differ (P > 0.10) from that of heifers fed 20AH/0WCGF or 30AH/0WCGF, and which had greater gain efficiencies (P < 0.05) than heifers fed 30AH/0WCGF. Rate of DMI increased linearly (P < 0.01) with increasing AH and decreased linearly (P < 0.01) with increasing WCGF. Heifers were finished on diets containing 33% WCGF with 0 or 0.5% added urea (DM basis) offered ad libitum. Increasing WCGF in growing diets tended (linear, P < 0.10) to increase finishing ADG and gain efficiency, whereas increasing AH decreased (linear, P < 0.05) kidney, pelvic, and heart fat, and the percentage of carcasses grading USDA Prime. Urea tended to increase ADG (P < 0.10), but decreased (P < 0.04) the percentage of carcasses grading USDA Choice. Results suggest that the value of WCGF relative to steam-flaked corn in limit-fed growing diets might be improved in diets containing 30% AH relative to diets containing 10 or 20% AH.

Isolation and characterization of pokeweed antiviral protein mutations in Saccharomyces cerevisiae: identification of residues important for toxicity.

Pokeweed antiviral protein (PAP), a 29-kDa protein isolated from Phytolacca americana inhibits translation by catalytically removing a specific adenine residue from the 28S rRNA of eukaryotic ribosomes. PAP has potent antiviral activity against many plant and animal viruses, including human immunodeficiency virus. We describe here development of a positive selection system to isolate PAP mutants with reduced toxicity. In vitro translation in the presence or absence of microsomal membranes shows that PAP is synthesized as a precursor and undergoes at least two different proteolytic processing steps to generate mature PAP. The PAP cDNA was placed under control of the galactose-inducible GAL1 promoter and transformed into Saccharomyces cerevisiae. Induction of PAP expression was lethal to yeast. The PAP expression plasmid was mutagenized and plasmids encoding mutant PAP genes were identified by their failure to kill S. cerevisiae. A number of mutant alleles were sequenced. In one mutant, a point mutation at Glu-177 inactivated enzymatic function in vitro, suggesting that this glutamic acid residue is located at or near the catalytic site. Mutants with either point mutations near the N terminus or a nonsense mutation at residue 237 produced protein that was enzymatically active in vitro, suggesting that the toxicity of PAP is not due solely to enzymatic activity. Toxicity of PAP appears to be a multistep process that involves possibly different domains of the protein.

Effects of wet corn gluten feed and intake level on diet digestibility and ruminal passage rate in steers.

Twelve ruminally cannulated Jersey steers (BW = 534 kg) were used in an incomplete Latin square design experiment with a 2 x 2 factorial arrangement of treatments to determine the effects of wet corn gluten feed (WCGF) and total DMI level on diet digestibility and ruminal passage rate. Treatments consisted of diets formulated to contain (DM basis) steam-flaked corn, 20% coarsely ground alfalfa hay, and either 0 or 40% WCGF offered once daily for ad libitum consumption or limited to 1.6% of BW (DM basis). Two consecutive 24-d periods were used, each consisting of 18 d for adaptation, 4 d for collection, and a 2-d in situ period. Rumens of all steers were evacuated once daily at 0, 4, 8, and 12 h after feeding. Chromic oxide (10 g/[steer*d]) was fed as a digestibility marker, and steers were pulse-dosed with Yb-labeled alfalfa hay to measure ruminal particulate passage rate. Dacron bags containing 5 g of steam-flaked corn, WCGF, or ground (2-mm screen) alfalfa hay were placed into the rumens of all steers and removed after 3, 6, 12, or 48 h. Wet corn gluten feed increased percent apparent total-tract digestion of OM (P < 0.01), NDF (P < 0.01), and starch (P < 0.03), decreased (P < 0.01) ruminal total VFA concentration, increased (P < 0.01) ruminal NH3 concentration, and increased (P < 0.01) ruminal pH. Wet corn gluten feed also increased (P < 0.01) ruminal passage rate of Yb. Limit feeding decreased (P < 0.01) percent apparent total-tract digestion of both OM and NDF, ruminal total VFA concentration (P < 0.01), and ruminal fill (P < 0.01), but increased (P < 0.01) ruminal NH3 concentration. Apparent total-tract digestion of starch was not affected (P = 0.70) by level of DMI. A DMI level x hour interaction (P < 0.01) occurred for ruminal pH. Limit feeding increased ruminal pH before and 12 h after feeding, but decreased ruminal pH 4 h after feeding compared with diets offered ad libitum. A diet x DMI level interaction (P < 0.02) occurred for in situ degradation of alfalfa hay, with dietary addition of WCGF increasing (P < 0.02) the extent of in situ alfalfa hay degradation in steers fed for ad libitum consumption. This study suggests that WCGF increases OM and NDF digestion, and that limit feeding diets once daily might depress OM and NDF digestion, possibly due to decreased stability of the ruminal environment.