Characterization of a repetitive DNA probe for Babesia bigemina

A plasmid (p16) containing a Babesia bigemina DNA insert was selected and labeled with 32P. This probe was evaluated for specificity and sensitivity by dot blot hybridization. The probe was specific and hybridized with only Babesia bigemina DNA, and not DNA from Babesia bovis, bovine leukocyte, Trypanosoma brucei or Anaplasma marginale. The DNA probe detected as little as 10 pg of Babesia bigemina DNA. The probe hybridized with Babesia bigemina isolates from Mexico, the Caribbean region and Kenya. Genomic Babesia bigemina DNA of a Kenyan isolate was digested with restriction endonucleases, and the fragments were separated by gel electrophoresis and Southern blotted. The filter was hybridized with labeled p16 and each endonuclease digestion produced at least 16 resolvable DNA fragments. The inserted Babesia bigemina DNA was approximately 6.3 kb in size. A partial restriction map was constructed. A simple whole blood dot blot procedure was utilized to evaluate the sensitivity of the DNA probe. This probe would detect as few as 150 Babesia bigemina infected erythrocytes contained in a 1-microliter sample. The DNA probe has the potential to be a very sensitive and specific diagnostic tool.     

Comparing the detection of exposure to Ehrlichia ruminantium infection on a heartwater-endemic farm by the pCS20 polymerase chain reaction assay and an indirect MAP1-B enzyme linked immunosorbent assay

Detection of heartwater is not always easy especially because all the serological assays so far available either have poor sensitivity or specificity. The indirect MAP-1B ELISA has been reported to be the most specific test for heartwater, although it does also detect antibodies to some closely related ehrlichial agents. This study was undertaken to compare two methods for the detection of heartwater infection caused by the ehrlichial agent Ehrlichia (Cowdria) ruminantium. Fifteen cattle on a heartwater-endemic farm infested with high numbers of Amblyomma hebraeum ticks, and hence exposure to E. ruminantium infection were monitored over an 8-week period by pCS20 PCR and an indirect MAP-1B ELISA. Infection was detected by pCS20 PCR in most animals with the highest number of positives (60%) in week 6 of the study. Similarly, exposure to E. ruminantium was detected by indirect MAP-1B ELISA in some animals, with the highest number of seropositives (27%) at weeks 2-6 of the study. The data demonstrated a fluctuating rickettsaemia in cattle in a heartwater-endemic area. Comparison of the two tests indicated that the pCS20 PCR assay was more reliable because it detected more infections than the indirect MAP-1B ELISA and would therefore be the method of choice for detection of E. ruminantium infection.     

Effect of dietary selenium and vitamin E on spermatogenic development in boars

An experiment involving a total of 61 crossbred boars evaluated the effects of dietary Se and vitamin E on spermatogenic development at various stages of sexual development and the prostaglandin F2alpha (PGF2alpha) content in the seminal vesicle and prostate glands at 18 mo of age. The experiment from 5.4 to 9 mo of age was conducted as a 2 x 2 factorial in a randomized complete block design. Dietary Se at 0 or .5 ppm was the first factor and vitamin E at 0 or 220 IU/kg diet was the second. From 9 to 18 mo of age, a group of sexually active and inactive boars was a third factor. Treatment diets were fed from weaning (28 d of age) to the end of the experiment. Three boars per treatment group at 5.4 (105 kg BW), 6.2 (130 kg BW), and 9.0 (150 kg BW) mo of age were killed and the testes collected. From 9 to 18 mo of age, three boars from each dietary treatment group were used for semen collection, and another set of three to four boars from each treatment group remained sexually inactive. At 18 mo, both sets of boars were killed and their testes, prostates, and seminal vesicles were collected. The testis at each age was evaluated for sperm reserve numbers and germ and Sertoli cell populations. At 5.4 or 6.2 mo of age, testicular sperm reserves were not affected by dietary Se (P > .15), at 9.0 mo of age there was a trend for a higher (P < .10) number of sperm reserves, and by 18 mo of age the Se-fed boars had higher (P < .01) numbers of sperm reserves. Vitamin E had no effect (P > .15) on testicular sperm reserves at any age period. Boars fed dietary Se had a greater number of Sertoli cells (P < .01) and round spermatids (P < .01) at 6.2 mo of age, but by 18 mo of age the boars fed Se had more Sertoli cells (P < .05), more secondary spermatocytes (P < .01), and more round spermatids (P < .05). Vitamin E did not affect Sertoli or germ cell populations at the various ages. Boars at 18 mo of age had lower PGF2alpha concentrations in the prostate (P < .05) and seminal vesicles (P < .01) when vitamin E was fed, whereas Se had no effect. Sexually active boars had lower PGF2alpha concentrations in the seminal vesicles (P < .01) than sexually inactive boars, but there was no effect (P > .15) of sexual activity on the number of Sertoli cells, primary or secondary spermatocytes, or round spermatids. Our results indicate that Se has a role in establishing the number of boar spermatozoal reserves and Sertoli cells, whereas supplemental vitamin E did not affect these criteria.     

Effect of feed intake during late development on pubertal onset and resulting body composition in crossbred gilts.

A total of 105 nonboar-exposed, F2 ([Landrace x Yorkshire] x Duroc) gilts were used in two replicates of a randomized complete block experiment to evaluate the effect of dietary feed intake on pubertal onset and subsequent body composition. Feed intakes were established at 50% of ad libitum (AL-50), 75% of ad libitum (AL-75), or at ad libitum (AL-100) levels from 4.5 to 9 mo of age. A corn-soybean meal diet fed to all gilts was formulated to meet or exceed nutrient requirements except for energy. Puberty was measured by two methods: 1) monitored once daily by back pressure applied by the herdsman or 2) from elevated plasma progesterone concentrations. Body composition was evaluated by the deuterium oxide method after plasma progesterone concentrations were elevated. Daily feed intake for the experimental period averaged 1.6, 2.3, and 3.2 kg, and the BW of gilts at 8 mo of age were 111, 131, and 154 kg for the AL-50, AL-75, and AL-100 groups, respectively. Body weight, backfat thickness, and body fat content increased linearly (P < .01) as feed intake increased, but age at puberty was not severely influenced. A minimum body fat content or percentage did not seem to initiate pubertal onset. There was a trend for a lower percentage of the AL-50 gilts to ovulate (P = .08) than those fed the AL-75 and AL-100 intakes. An inverse relationship resulted between the percentage of gilts that ovulated to the percentage that showed behavioral estrus.(ABSTRACT TRUNCATED AT 250 WORDS)     

Effect of organic and inorganic selenium sources and levels on sow colostrum and milk selenium content

A study was conducted to evaluate the short-term effects of feeding two dietary Se sources at various Se levels on the transfer of Se to the dam's milk and nursing pig. Six dietary treatments were arranged in a 2 x 2 factorial arrangement with two additional treatments in a randomized complete block designed experiment. Inorganic (sodium selenite) or organic (Se-enriched yeast) Se sources were added to the diet at .15 or .30 ppm Se. A non-Se-fortified corn-soybean meal basal diet served as a negative control, and a sixth group was fed .15 ppm Se from both inorganic and organic Se sources. A total of 43 sows were fed their treatment diets at 2.2 kg/d from 6 d prepartum to parturition and at full feed through a 14-d lactation period. Ten sows were initially bled at 6 d prepartum, and three sows and three pigs from their litters were bled at 7 and 14 d postpartum. Serum was analyzed for its Se concentration and glutathione peroxidase (GSH-Px) activity. Colostrum was collected within 12 h postpartum and milk at 7 and 14 d of lactation. When the basal diet was fed, sow serum GSH-Px activity declined from 6 d prepartum and remained low throughout lactation. When dietary Se levels increased, sow serum Se concentration and serum GSH-Px activity increased (P < .05) at both 7 and 14 d postpartum. The short-term feeding of either Se source at .15 or .30 ppm Se did not affect colostrum Se content when inorganic Se was fed, but it was increased when organic Se was provided. This resulted in a significant Se source x Se level interaction (P < .01). Milk Se at 7 and 14 d postpartum was 2.5 to 3 times higher when the organic Se source was provided and resulted in a significant Se source x Se level interaction (P < .05). When the combination of inorganic and organic Se was fed at .15 ppm Se, colostrum and milk Se contents were similar to those of sows fed .15 ppm Se from the organic Se source. Pig serum GSH-Px activity was not affected at 7 and 14 d of age by dietary Se level or Se source fed to the sow, but serum Se increased (P < .05) as dietary Se level increased, particularly when sows had been fed organic Se. The results demonstrated that organic Se increased milk Se content more than did inorganic Se and increased the nursing pig's serum Se. These results indicate that inorganic Se was more biologically available for sow serum GSH-Px activity, but organic Se was more effectively incorporated into milk.     

Biological characteristics for assessing low phosphorus intake in growing swine

A total of 195 swine from 18 to 35 kg were used to evaluate various biological responses of growing swine fed low P diets at different Ca:P ratios. Three replicates of a randomized complete-block design in a 3 X 3 factorial arrangement of treatments were used. Total dietary P levels of .12, .31 and .50% were fed in 1:1, 1.5:1 or 3:1 Ca:P ratios. Diets were formulated using corn starch, dextrin and soy protein concentrate, with monosodium phosphate and limestone used to supply appropriate dietary treatment levels of Ca and P. Gain and feed performance were generally influenced by both Ca:P ratio and dietary P level. Serum inorganic P decreased linearly as Ca:P ratio widened and increased linearly as dietary P increased. Serum Ca concentrations responded inversely to serum inorganic P in response to both Ca:P ratio and dietary P level. Serum alkaline phosphatase was not affected by Ca:P ratio, but declined quadratically with increasing dietary P level. Bone bending moment was influenced by both dietary P level and Ca:P ratio. Bone component weights (ash, organic matrix), percentage bone ash and net accretion of bone ash and organic component weights were not influenced by Ca:P ratio but increased linearly as dietary P level increased. These results suggest that bone component weights, net ash accretion and percentage bone ash were more sensitive criteria than bone bending moment or serum alkaline phosphatase, which in turn were more sensitive than serum inorganic P, Ca, Mg and growth performance characteristics in response to dietary P level.     

Effect of dietary selenium source, level, and pig hair color on various selenium indices

The first experiment evaluated the effects of feeding various levels of Se, two Se sources, and hair color on selenosis responses in growing-finishing pigs. The study conducted in two replicates was a 2 x 6 x 2 factorial arrangement in a split-plot design. Sodium selenite and Se-enriched yeast added at 0.3, 1, 3, 5, 7, and 10 ppm Se served as the main plot and pig hair color as the subplot. A total of 96 crossbred pigs were allotted and fed their treatment diets for a 12-wk period. White and dark (red or black) hair samples were collected from the dorsal-midline at the 4-, 8-, and 12-wk periods from one pig of each hair color from each treatment pen. Lower pig weights (P < 0.10) and daily gains (P < 0.05) occurred as dietary Se level increased when pigs were fed either Se source. Selenosis responses were somewhat more severe, when the inorganic Se source was fed. Alopecia and hoof separation were encountered after the 8-wk period when pigs were fed inorganic rather than organic Se. Plasma Se increased as dietary level increased (P < 0.01), when organic Se was provided (P < 0.01), and was higher (P < 0.05) when pigs were white-haired. A time x hair color x dietary Se level interaction (P < 0.05) occurred, in which hair Se concentration was higher in dark- than in white-colored pigs and increased as dietary Se level increased as the experiment progressed. The correlation coefficient between dietary Se level and hair Se concentration averaged 0.90 (P < 0.01). Cysteine was the amino acid in the highest concentration in hair, but this and other amino acids were not affected by Se level, Se source, or hair color. A second experiment was a 3 x 6 factorial arrangement in a split-plot design with three 9-mo-old gilts from each of the Yorkshire, Duroc, and Hampshire breeds to determine whether hair Se concentration differed by body location and breed. Hair samples were collected from the shoulder, back, rump, front-leg, belly, and hind-leg areas. Hair Se concentration was higher in red- and white-haired pigs and lower in black-haired gilts (P < 0.01). Higher hair Se concentrations (P < 0.05) occurred from the lower than from the upper body areas. Our results suggest that selenosis occurs at dietary levels > 5 ppm and that white-haired pigs exhibit alopecia sooner than dark-haired pigs. No difference in hair Se concentration occurred when diets were < 1 ppm Se, but as dietary Se level increased dark-haired pigs retained more Se in their hair than white-haired pigs.     

Deficit irrigation in a production setting: canopy temperature as an adjunct to ET estimates

Water available for agricultural use is declining worldwide as a result of both declining water resources and increasing application costs. Managing crop irrigation under conditions where the water need cannot be fully met represents the future of irrigation in many areas. On the southern high plains of Texas there is interest among producers to reduce the amount of water applied to cotton. In this study, a producer’s efforts to reduce water application to a cotton crop were assessed in terms of a comparison between evapotranspiration, rainfall, and irrigation that is widely used in the region. The producer was able to reduce water application to meet intended reductions relative to the evapotranspiration estimates but, depending on the method used for calculating the crop water need, he tended to over water the crop in two out of three intended deficit irrigation regimes. Analysis of continuously monitored canopy temperatures provided verification of over-irrigation. Continuously monitored canopy temperature is proposed as a useful adjunct to evapotranspiration approaches to deficit irrigation management.