Isolation and purification of a diagnostic antigen for bovine cysticercosis by hydrophobic chromatography

An ammonium sulfate fraction of Taenia hydatigena cyst fluid (ThFAS) was further fractionated by hydrophobic interaction chromatography, using alkylagarose and omega-amino alkylagarose columns, in an effort to isolate and purify a specific diagnostic antigen in the ThFAS preparation. The less than 12 kDa antigen was found to have an affinity for immobilized alkanes with chain length of six carbons or greater. The antigen was recovered in an ethylene glycol eluate from a hexylagarose column then analyzed by Western blot; it reacted with bovine and human cysticercosis infection sera and with specific monoclonal antibodies but not with control sera or fascioliasis infection sera. When the eluate was used as coating antigen in a plate ELISA assay no false positive reactions were seen in sera from cattle infected with Fasciola hepatica; false positive reactions were observed for the unfractionated ThFAS antigen preparation.     

Importance of Some Biochemical Parameters in the Quality and Yield of Rapeseed (Brassica napus L.)

Samples of rapeseed from three Italian growing environments (Bologna, Perugia and Palermo) were analysed for glucose content and dry weight of 1000 seeds every three or four days starting from the end of flowering until complete ripening. In addition, the content of oil, soluble and total proteins, glucosinolates and myrosinase activity was determined in samples of mature seeds. The cultivars used were jet Neuf and Lingot (type 0) and Tandem, Jade and Santana (type 00). From the results it emerged that the point of intersection of the two branches of the linear regression plots for different glucose-consumption kinetics found during seed filling, in addition to being strongly affected by the climate of the test environment, is correlated with quantitative and qualitative production, independently of the genotype.     

Accuracy of the thermodilution method in estimating high flow - an in vitro study

The accuracy of thermodilution for measuring flow rates of 10–40 L/min was evaluated using a commercially available thermodilution cardiac output computer in an in vitro model. Water (36.5–37.5°C) was directed through a mixing chamber via a constant flow pump. Thermodilution estimates of flow using four different volumes (10, 20, 30, 40 ml) of iced water injectate were compared to simultaneous measurements of timed samples of effluent from the mixing chamber. Injectate volume had a significant impact on the accuracy of thermodilution estimation (p < 0.05). Thermodilution overestimated measured flow when 10 and 20 ml of injectate were used to determine flow rates < 20 L/min but underestimated flow when injectate volumes of 30 and 40 ml were used, or when measured flow was > 25 L/min. The discrepancy between thermodilution flow and measured flow increased as rate of fluid flow increased.     

Variation in the level of grain defect light flecks and spots on cattle hides.

The occurrence of hide damage light flecks and spots was determined on tanned hides from 28 herds during a period of 8 to 12 months. Light flecks and spots are described as small areas of grain loss up to 3 mm in diameter that are seen on dyed crust cattle leather. Damage was found on 75.8% of all hides. The neck and shoulders were the anatomical region with the highest prevalence of damage. Sixty-eight per cent of all hides had light flecks and spots in this region. The forelimbs and dewlap were the anatomical region with the second highest occurrence with a prevalence of 39.1%. This distribution corresponded to the known distribution of lice in cattle. No significant differences were observed in age, sex, prevalence of lice in the herd assessed in March or infestations with different lice species. The frequency of light flecks and spots varied significantly during the year. The frequency was highest in the late winter and early spring, decreased significantly during the summer and was lowest in the autumn. This variation supported the importance of lice in the development of light flecks and spots and suggested a relatively long healing period for the damages induced by lice.     

Sources of error with in vitro digestibility assay of pasture feeds

A series of experiments was undertaken to determine population statistics for in vitro organic matter digestibility ( in vitro OMD) data and to examine the effects of basal diet, donor animal and precollection fasting interval on the activity and specificity of rumen fluid inoculum. The experiments utilized wether sheep, a diverse set of pasture grass and legume feeds prominent in the Australian subtropics and the Tilley and Terry in vitro digestibility procedure running under the operating pressure of a practicing feeds evaluation laboratory.
The standard errors of in vitro OMD estimates for within and between batch runs were ±0·88 × 10⁻² and 0·62 × 10⁻², respectively. These error terms were used to develop protocols to accept, reject or scale raw in vitro OMD data. Differences between donor animals in the activity of rumen fluid were highly significant. Extending the precollection fasting interval beyond 16 h was associated with a substantial decline in inoculum activity.
An in vitro-in vivo calibration relationship based on fifteen test feeds and using lucerne ( Medicago sativa ) as basal diet was described by the linear model y = 1·3 x-0·195±4·9 × 10⁻² r = 0·79 (y = in vivo OMD, x = in vitro OMD). Despite large effects of basal diet on both the absolute values and relative ranking of test feeds, neither the RSD nor r values were improved using alternative diets to Lucerne chaff.
The results highlight the need to formally standardize the analytical and biological components of the in vitro digestibility procedure to safeguard the integrity of data.     

Free-running rhythms of adrenocorticotropic hormone (ACTH), cortisol and melatonin in pigs.

In the domestic pig, a circadian rhythm of plasma cortisol occurs, with greatest concentrations in the morning and lowest concentrations in the afternoon. However, photic entrainment of the rhythms of ACTH and melatonin in pigs have not been defined clearly. This experiment was designed to evaluate free-running rhythms of ACTH, cortisol and melatonin in pigs housed in constant light (LL) and constant darkness (DD). Twelve crossbred barrows, maintained under ambient photoperiod, were catheterized and tethered individually in two environmentally controlled rooms, one with LL and the other with DD. For animals in LL, fluorescent lights provided 202 +/- 15 (mean +/- standard deviation) lux of light at 65 cm above the floors. Incandescent nightlights equipped with 7 watt red bulbs provided 7 +/- 2 lux and were illuminated continuously in both rooms. Pigs were given at least 14 d exposure to LL and DD, then samples of plasma and serum were obtained at hourly intervals for 48 hr. Plasma was assayed for ACTH, and serum for cortisol and melatonin. Periodograms were constructed to analyze the data. For this type of analysis, a statistic, Qp, is calculated, and circadian periodicity is suggested if maximum Qp (Qp max) occurs at or near 24 hr. The period of the free-running rhythms (tau) at Qp max for ACTH, cortisol and melatonin for pigs in LL (23.80 +/- .01, 23.78 +/- .01, and 23.21 +/- .02 hr, respectively) did not differ significantly from those for pigs in DD (23.39 +/- .01, 23.20 +/- .01, and 22.55 +/- .02 hr, respectively).(ABSTRACT TRUNCATED AT 250 WORDS)     

Detection and characterization of leptospiral antigens using a biotin/avidin double-antibody sandwich enzyme-linked immunosorbent assay and immunoblot.

A biotin/avidin double-antibody sandwich enzyme-linked immunosorbent assay (ELISA) for the detection of antigens of Leptospira interrogans serovars in experimentally inoculated bovine urine samples was evaluated. Immunoglobulin G (IgG) from rabbits immunized with L. interrogans serovar hardjo type hardjobovis sonicated, whole cell, and formalinized-heated antigen preparations were purified by a protein A-superose column coupled to fast protein liquid chromatography, and evaluated for species specificity in the ELISA. The ELISA using each specific IgG detected as few as 10(4) leptospires of the homologous serovar hardjo diluted in phosphate-buffered saline solution with Tween 20 (PBSS-Tween 20). On immunoblot analysis of proteinase-K-digested whole cell leptospiral preparations, each IgG revealed the presence of bands specific to serovar hardjo, suggesting the presence of serovar-specific epitopes on the lipopolysaccharide molecules. The minimum number of cells of heterologous serovars pomona, grippotyphosa, bratislava, icterohaemorrhagiae and copenhageni detected by each ELISA was greater, ranging from 10(6) to 10(7). The common antigenic determinants observed on immunoblot analysis were different for each specific IgG, except for a major cross-reacting, possibly flagellar, protein doublet at approximately 36-36.5 kDa. Leptospires were equally well detected by the ELISA in both bovine urine and PBSS-Tween 20.