A methodology to detect and quantify five pathogens causing potato tuber decay using real-time quantitative polymerase chain reaction

ABSTRACT Late blight (Phytophthora infestans), pink rot (Phytophthora erythroseptica), leak (Pythium ultimum), dry rot (Fusarium sambucinum), and soft rot (Erwinia carotovora subsp. carotovora and subsp. atroseptica) are particularly damaging diseases of stored potato tubers worldwide. In this study, we present a methodology to detect and quantify the causal agents of the five aforementioned diseases from whole potato tubers, using real-time quantitative-polymerase chain reaction. Six primer pairs were designed to amplify targets smaller than 150-bp DNA in single copy protein-coding gene targets of each of the pathogens and the potato host. Using a large collection of pure culture DNA samples, all primer pairs specifically detected the DNA target in the intended pathogenic species. Amplification efficiencies over a five-log dilution series ranged between 95 and 100% and were unaffected by the presence of large amounts of host DNA. The detection level of the primers reached 0.5 pg of target DNA. Pathogens were detected in 100 pg of total DNA extracted from 170 to 250 g of tubers, 4 days after inoculation, regardless of the presence of symptoms. The presence of P. erythroseptica, Pythium ultimum, or E. carotovora was also detected in 1 ng of DNA extracted from potato tubers collected from a commercial storage facility. This study provides the first step in a methodology to predict the storability of potato tubers following harvest.     

Effects of diets of varying energy concentrations on the growth and carcase composition of geese

Two hundred and eighty eight-d-old goslings were allocated at random to one of 18 groups, each group comprising 8 of each sex. Three starter and three grower diets (both 11, 12, 13 MJ AME/kg) were fed and were combined in a factorial design to give 9 treatments. The starter diets were given for 4 weeks and the grower diets from weeks 5 to 9 inclusive. Body weight was unaffected by treatment, but the males were significantly heavier than the females. Food intake declined significantly as the dietary energy concentration increased. The efficiency of food utilisation of birds fed on the starter diet with the lowest energy concentration was significantly less than that of birds fed on the other diets. Birds fed on the grower diet with the highest energy concentration gave significantly better FCE than those fed on the other diets. There was no evidence to suggest that the treatments had any significant effect on carcase components and chemical composition of the geese. Oleic acid was significantly depressed and linoleic acid concentration increased as the energy concentration of the diets increased. However, this may reflect the higher fat concentrations in the diets with the higher energy concentrations.     

Body growth and testicular characteristics of boars fed a synthetic progestogen, altrenogest

A synthetic progestogen (altrenogest) was fed to 24 Yorkshire X Duroc boars to determine effects on body growth, serum testosterone, and testicular characteristics. Boars from six litters (blocks) were allotted randomly to one of four treatment groups at 12 wk of age. Treatment groups were: controls, altrenogest fed for 3 wk, altrenogest fed for 6 wk and altrenogest fed for 9 wk. Treatment began at 15 wk of age at a daily dose of 20 mg X boar-1. Although there were no differences among treatment groups for gain and feed intake, boars fed altrenogest for 6 and 9 wk tended to consume more feed and were less (P less than .05) efficient than boars fed altrenogest for 3 wk or controls. Boars fed altrenogest for 3, 6 and 9 wk maintained lower (P less than .05) peripheral serum testosterone concentrations than controls from 15 to 24 wk of age. However, serum testosterone increased after altrenogest withdrawal in the 3- and 6-wk treatment groups but did not reach control concentrations by 24 wk. Boars fed altrenogest for 9 wk maintained serum testosterone below 1 ng/ml during the treatment period. Despite lower concentrations of serum testosterone in altrenogest-treated boars, backfat thickness was similar to controls, perhaps suggesting a slight anabolic effect of altrenogest on nutrient partitioning. Testicular weights and volumes at 24 wk decreased (P less than .001) linearly with increased duration of altrenogest feeding. Serum testosterone and estradiol in testicular venous effluent at castration were lower (P less than .01) in altrenogest-treated boars than in controls. These data demonstrate that feeding altrenogest inhibits both testicular growth and steroidogenesis of boars without altering body growth or backfat thickness.     

Duration of the persistent activity of moxidectin against Haemonchus contortus in sheep

SUMMARY Weaned lambs were infected with Haemonchus contortus 35,28,21,14 and 7 days after treatment with moxidectin at 0.2 mg/kg and 35 and 14 days after treatment with ivermectin at the same dose rate. Worm counts 14 days after infection showed that moxidectin prevented the establishment of over 99% of infective larvae for 28 days and reduced the establishment rate at 35 days by 96%, relative to ivermectin. There was no difference in the protective efficacy of ivermectin at 14 or 35 days. The persistence of moxidectin is likely to provide advantages in nematode control, particularly when used as a strategic early summer treatment or as a pre-lambing treatment to ewes. Implications of the persistent activity of moxidectin for the development of resistance during the decay phase are discussed.     

Reproductive traits, lactation and foal growth in mares fed altrenogest

Lactating mares were assigned as controls or fed altrenogest (.044 mg.kg body wt-1.d-1) for 15 d after foaling. Mares (n = 6) fed altrenogest were inseminated during the first estrus after treatment and mares (n = 6) in the control group were inseminated during the second postpartum estrus. Ovulation during the estrus in which mares were inseminated occurred 26 +/- 1 d postpartum for treated mares and 36 +/- 1 d postpartum for control mares. The percentage of mares conceiving was not different for control (67%) and alternogest-treated (100%) mares. No differences were observed in tone and size of the uterus or size of the ovulatory follicle between treated and control groups. Uterine cultures and biopsies collected on d 7 and 15 postpartum were similar between treatment and control groups in bacterial populations or endometrial epithelial cell height. Blood was collected on d 7, 11, 15, 19 and 23 postpartum, and concentrations of estradiol-17 beta in serum were determined by radioimmunoassay. Mean concentrations of estradiol-17 beta across days were 10 +/- .8 and 12 +/- .6 pg/ml for control and treated mares, respectively. Concentrations of serum estradiol-17 beta were higher (P less than .05) in treated mares on d 23 postpartum. Daily milk yields, determined by the weigh-suckle-weigh method, and milk composition were similar between treatment groups on each collection day. Altrenogest can be used to predictably delay estrus in the postpartum mare without altering fertility, yield and composition of milk, or foal growth.     

Effect of energy intake after weaning on the sexual development of beef bulls. II. Age at first mating, age at puberty, testosterone and scrotal circumference

Simmental and Hereford bulls were individually fed varying levels of the same diet to determine the effect of energy intake after weaning on rate of sexual development. For 200 d, 29 Simmentals were fed an average of 14.6, 19.2 or 23.8 Mcal and 27 Herefords were fed 13.4, 17.5 or 22.2 Mcal metabolizable energy per bull daily. Higher energy did not hasten sexual development, as measured by serum testosterone, age at first mating or age at puberty. As energy level increased, weight at puberty increased (P less than .05) for both breeds, and scrotal circumference at puberty increased (P less than .05) for Simmentals. When adjusted for breed and energy level, bulls that were heavier at 1 yr of age had larger scrotal circumference (P less than .05) and were younger at puberty (P less than .01). Bulls that had higher serum testosterone at 1 yr of age reached puberty earlier (P less than .01). At a constant age there was no relationship between testosterone and scrotal circumference. Although higher levels of energy increased scrotal circumference of Simmentals, it did not hasten sexual development for either breed.