An ultrasonographic scoring method for transabdominal monitoring of ascarid burdens in foals.

REASONS FOR PERFORMING STUDY: Parascaris spp. infections can lead to life-threatening small intestinal impactions in foals. Currently available diagnostic techniques cannot estimate the magnitude of an ascarid burden, and hence identify foals potentially at risk of developing impactions. OBJECTIVES: To describe and evaluate an ultrasonographic transabdominal scoring technique for monitoring of ascarid burdens in foals and to perform a cost-benefit analysis of the application of this technique. STUDY DESIGN: A transabdominal ultrasonographic technique was validated against ascarid worm counts from 10 foals aged 162-294 days. In a treatment trial, 15 foals were randomly allocated to 3 treatment groups: ivermectin, oxibendazole and no treatment. Blinded ultrasound examinations were performed daily for 5 consecutive days following treatment. Foals were examined ultrasonographically twice by the same investigator, and by different investigators for intra- and interobserver agreement evaluation. Cost-benefit analyses identified threshold values for the probability of ascarid impactions above which the screening method becomes cost-effective. METHODS: The ultrasound technique used 3 locations along the ventral midline. An ascarid scoring system was established that assessed the magnitude of ascarid burden ranging from 1-4. The method was validated against worm burdens of 10 worms and above with calculation of diagnostic specificity, sensitivity, and predictive values. Treatment trial data were evaluated statistically using mixed model analysis. Kappa values were generated for intra- and interobserver agreement. RESULTS: Two consecutive examinations were found to detect worm burdens >10 ascarids reliably. Ascarid scores declined in response to both anthelmintic treatments, although differences were not statistically significant. Kappa values indicated fair to moderate intra- and interobserver agreements. The majority of cost-benefit analyses indicated that ultrasound examinations are cost effective when the probability of ascarid impactions is above a range of 0.0001-0.0082 (i.e. 1 in 10,000 to 8 in 1000 foals). CONCLUSIONS: The ultrasonographic screening techniques can be a useful tool for monitoring ascarid burdens in foals.

Biphasic appearance of corticated and decorticated ascarid egg shedding in untreated horse foals.

Parascaris spp. infects foals worldwide and may cause airway inflammation in addition to small intestinal impaction and rupture. It is observed that acquired immunity eliminates ascarid burdens beginning at about 6 months of age, and current evidence suggests that a single parasite generation propagates in each foal crop. The purpose of this study was to monitor natural parasitic infections in untreated mixed breed horse foals over the course of 0-300 days of age. Fecal samples were collected monthly from all foals born in 2014 (n=13), beginning July 2014 through February 2015. Fecal egg counts (FECs) were performed in triplicates using the Mini-FLOTAC method. The foals were necropsied between 154 and 298 days of age and all intestinal ascarid were collected and identified to stage. Ascarid FECs exhibited a biphasic distribution with an initial peak at 91-120 days of age and, after a steady decline, a second, smaller peak at 241-300 days of age. Numbers of corticated and decorticated ascarid eggs were compared, with decorticated FECs remaining consistently low with a slight increase directly after the first corticated FEC peak. Overall, 4.36% of the total ascarid eggs counted were decorticated. Ascarid FECs showed a sharp peak in September, declined, and then steadily increased beginning in December and continuing through February. Upon necropsy, moderate to high number of ascarid specimens were recovered from foals between 8 and 10 months of age, coinciding with the second peak for the FECs. Eleven of the 13 foals harbored immature ascarid stages indicating a recent reinfection. However, these data demonstrates that apparently a second, smaller wave of infection is present in 8-10 month old foals. It may be of value to monitor egg counts in this age group to make sure that all parasite categories are well controlled.

The development of the Indian vision function questionnaire: questionnaire content.

AIM: To elicit problem statements describing the consequences of vision impairment as a first step towards the development of a vision related quality of life instrument for use in India METHODS: 46 focus groups were conducted in three regions of India. Separate focus groups were held for men and women and according to disease categories: cataract (24), glaucoma (six), diabetic retinopathy or macular degeneration (10), and "mixed low vision" (six). Facilitators followed a topic guide and sessions were audio taped and transcribed. Problem statements were extracted and coded and summarised into major problem domain areas. RESULTS: Nearly 5000 problem statements, an average of 15 statements per participant, were consolidated into 18 broad domain areas. The most important problem areas accounting for over 50% of all statements were ambulation, household or occupational activities, vision symptoms, and people recognition. A further quarter of statements related to difficulties with eating and drinking, psychological concerns, self care, reading, and watching television. Problem statements were similar across the disease groups, although rankings varied. CONCLUSIONS: The functional and psychological impacts described by visually impaired participants in India are similar to those reported in other population settings although the context and impact of problems vary.

Identification and characterization of an mRNA encoding a proline-rich protein that rapidly declines in abundance in the tips of harvested asparagus spears.

We previously isolated a cDNA clone, pTIP13, whose homologous mRNA rapidly declined in abundance in the tips of harvested asparagus (Asparagus officinalis L.) spears [King and Davies (1992) Plant Physiol. 100: 1661]. In order to identify factors regulating the postharvest deterioration of asparagus, we have now sequenced the pTIP13 cDNA, derived the encoded amino acid sequence and determined the cellular location of pTIP13 mRNA by in situ hybridization. pTIP13 encodes a derived protein that is rich in proline (22.3%), but also has a high content of lysine (15.2%) and threonine (14.1%). The proline residues are located in motifs at the amino-terminal region of the protein. The carboxyl-terminal region of the derived protein has a high leucine content and shares > 64% amino acid identity with derived proteins identified from cDNA clones to cell wall protein precursor mRNAs obtained from soybean hypocotyls, alfalfa roots, and tomato fruit. Genomic Southern analysis suggests that pTIP13 is encoded by a single-copy gene in asparagus. pTIP13 mRNA was localized to specific cell types in the young bracts of the asparagus spear tip. The results provide new information on the complexity of tissue responses in the tips of asparagus spears following harvest.

Polyuronides in Avocado (Persea americana) and Tomato (Lycopersicon esculentum) Fruits Exhibit Markedly Different Patterns of Molecular Weight Downshifts during Ripening.

Avocado (Persea americana) fruit experience a rapid and extensive loss of firmness during ripening. In this study, we examined whether the chelator solubility and molecular weight of avocado polyuronides paralleled the accumulation of polygalacturonase (PG) activity and loss in fruit firmness. Polyuronides were derived from ethanolic precipitates of avocado mesocarp prepared using a procedure to rapidly inactivate endogenous enzymes. During ripening, chelator (cyclohexane-trans-1,2-diamine tetraacetic acid [CDTA])-soluble polyuronides increased from approximately 30 to 40 [mu]g of galacturonic acid equivalents (mg alcohol-insoluble solids)-1 in preripe fruit to 150 to 170 [mu]g mg-1 in postclimacteric fruit. In preripe fruit, chelator-extractable polyuronides were of high molecular weight and were partially excluded from Sepharose CL- 2B-300 gel filtration media. Avocado polyuronides exhibited marked downshifts in molecular weight during ripening. At the postclimacteric stage, nearly all chelator-extractable polyuronides, which constituted from 75 to 90% of total cell wall uronic acid content, eluted near the total volume of the filtration media. Rechromatography of low molecular weight polyuronides on Bio-Gel P-4 disclosed that oligomeric uronic acids are produced in vivo during avocado ripening. The gel filtration behavior and pattern of depolymerization of avocado polyuronides were not influenced by the polyuronide extraction protocol (imidazole versus CDTA) or by chromatographic conditions designed to minimize interpolymeric aggregation. Polyuronides from ripening tomato (Lycopersicon esculentum) fruit extracted and chromatographed under conditions identical with those used for avocado polyuronides exhibited markedly less rapid and less extensive downshifts in molecular weight during the transition from mature-green to fully ripe. Even during a 9-d period beyond the fully ripe stage, tomato fruit polyuronides exhibited limited additional depolymerization and did not include oligomeric species. A comparison of the data for the avocado and tomato fruit indicates that downshifts in polyuronide molecular weight are a prominent feature of avocado ripening and may also explain why molecular down-regulation of PG (EC 3.2.1.15) in tomato fruit has resulted in minimal effects on fruit performance until the terminal stages of ripening.

Identification of peptides from autolysates of Saccharomyces cerevisiae that exhibit glucose tolerance factor activity in a yeast assay.

1. Cationic fractions were isolated from a low chromium (less than 0.2 ppm) commercial yeast extract in an attempt to purify the material responsible for glucose tolerance factor (GTF) activity observed in a standard yeast assay system. 2. Following previously described procedures a fraction with GTF activity but containing negligible chromium was isolated, which on further purification was found to be composed of many separate small basic peptides. 3. Much of the activity of the yeast GTF material in the yeast assay could be attributed to the presence of basic peptides and free amino acids acting as nitrogen sources for the yeast. 4. Additional activity was present in the yeast GTF sample, which was not due to a synergistic effect of the mixed amino acids and peptides although the component of the yeast extract responsible for this activity was not identified. 5. The results show that the GTF fractions isolated according to most previously published procedures are highly impure, and conclusions drawn about the nature of GTF based on these isolates must remain open to question. 6. The activity due to the presence of peptides and amino acids is a major cause of lack of specificity of the yeast systems as an assay for GTF.