Seroprevalence of Neospora caninum infection following an abortion outbreak in a dairy cattle herd

OBJECTIVE: To investigate the seroprevalence of Neospora caninum infection in a commercial dairy cattle herd, 15 months after detection of an abortion outbreak. PROCEDURE: Sera from the whole herd (n = 266) were examined for N caninum antibodies by indirect fluorescent antibody test (IFAT) and immunoblot analysis. Herd records were reviewed to collate serological results with abortion history, proximity to calving, and pedigree data. RESULTS: The seroprevalence of N caninum infection was 24% (63/266) for IFAT titre > or = 160, 29% (78/266) for immunoblot positive (+ve), and 31% (82/266) for IFAT > or = 160 and/or immunoblot +ve; 94% (59/63) of animals with IFAT > or = 160 were immunoblot +ve. The association between seropositivity (IFAT > or = 160 and/or immunoblot +ve) and history of abortion was highly significant (P < 0.001); the seroprevalence was 86% (18/21) in aborting cows, compared with 30% (50/164) in non-aborting animals. The abortion rate for seropositive cows was 26% (18/68) compared with 3% (3/117) for seronegative animals. IFAT titres of infected cows were higher within 2 months of calving than at other times (P < 0.001). The association between seropositivity in dams and daughters was highly significant (P = 0.009). CONCLUSIONS: The abortions were associated with N caninum infection and there was evidence of reactivation of latent infection close to calving and congenital transmission of infection. Immunodominant antigens identified by immunoblots may prove useful for improved diagnostic tests.     

Effects of dietary carbohydrates and buffering capacity on nutrient digestibility and manure characteristics in finishing pigs

A 2 x 3 factorial experiment was conducted with 24 finishing pigs (Yorkshire x [Finnish Landrace x Dutch Landrace]) to determine the effects of dietary buffering capacity (BC) and carbohydrate sources on apparent total tract digestibility (TD), N retention, and manure characteristics. Twelve of these pigs were fitted with steered ileo-cecal value cannulas to measure the apparent ileal digestibility (ID) of N. Experimental variables were two levels of BC (High = 600 mEq/kg and Low = 530 mEq/kg) and three dietary carbohydrates (tapioca [28%], soybean hulls [25%], and sugar beet pulp [25%]). The two levels of BC were achieved by adjusting the amounts of alkalogenic limestone and acidogenic Ca sulfate in the diet. Pigs were fed twice daily at 2.4 x maintenance requirement for metabolizable energy (418 kJ ME/BW(0.75)). Chromic oxide was used as an indigestible marker. Feces and urine were collected over 5 d for nutrient balance and in vitro measurements of pH and ammonia emission over 7 d. No significant interactions between BC and dietary carbohydrates on the ID, TD, N retention, or manure characteristics were observed. Lowering BC with Ca sulfate did not affect digestibility of most nutrients, except for a negative impact on the TD of Ca (P = 0.015) and Mg (P = 0.003). Although all pigs receiving Ca sulfate had more acidic urine (by 0.49 pH units; P = 0.001), ammonia emission from manure was not lowered, irrespective of the carbohydrate source. Carbohydrates affected significantly the ID and TD of most nutrients, whereas N retention remained similar (P > 0.10). Urinary N:fecal N ratios in manure of pigs fed diets with tapioca, soybean hulls, and beet pulp were 2.09, 1.35, and 1.67, respectively. These ratios corresponded with in vitro ammonia emission (169, 125, and 148 mmol/7 d; P = 0.023). In conclusion, these results indicate that the acidification of urine in pigs fed Ca sulfate in the presence of NSP-rich carbohydrates was achieved. However, acidity of manure and ammonia emission were not affected. The addition of NSP from soybean hulls and(or) sugar beet pulp reduced ratios of urinary:fecal N, and thereby ammonia emission.     

Variability among sources and laboratories in analyses of wheat middlings. NCR-42 Committee on Swine Nutrition

A cooperative research study was conducted by members of a regional committee (North Central Regional Committee on Swine Nutrition [NCR-42]) to assess the variability in nutrient composition (DM, CP, Ca, P, Se, NDF, and amino acids) of 14 sources of wheat middlings from 13 states (mostly in the Midwest). A second objective was to assess the analytical variability in nutrient assays among 20 laboratories (labs; 14 experiment station labs and six commercial labs). Wheat middlings were obtained from each participating station's feed mill. The bulk density of the middlings ranged from 289 to 365 g/L. The number of labs that analyzed samples were as follows: DM and CP, 20; Ca, 16; P, 15; Se, 7; NDF, 10; and amino acids, 9. Each lab used its own analytical procedures. The middlings averaged 89.6% DM, 16.2% CP, .12% Ca, .97% P, 36.9% NDF, .53 mg/kg Se, .66% lysine, .19% tryptophan, .54% threonine, .25% methionine, .34% cystine, .50% isoleucine, and .73% valine. As expected, there was considerable variation in nutrient composition among the 14 sources (P < .01), especially for Ca (.08 to .30%) and Se (.05 to 1.07 mg/kg). "Heavy" middlings (high bulk density, >335 g/L), having a greater proportion of flour attached to the bran, were lower in CP, lysine, P, and NDF than "light" middlings (<310 g/L), having cleaner bran, resulting in negative correlations between bulk density and CP (r = -.61), lysine (r = -.59), P (r = -.54), and NDF (r = -.81). Each 1-percentage-point increase in CP in the wheat middlings was associated with .0235 (r2 = .61) and 2.1 (r2 = .39)-percentage-point increases in lysine and NDF, respectively. Lysine content was associated with NDF, CP, and bulk density of wheat middlings (r2 = .88). There was considerable variation among laboratories (P < .01) in analysis of all nutrients. The CV among sources (100 x sigmaS/mean) was greater than among labs (100 x sigmaL/mean) for CP, Ca, P, Se, and NDF, but the CV among labs was greater than that among sources for DM and all of the amino acids except lysine and phenylalanine.     

Regeneration of Psychotria umbellata and production of the analgesic indole alkaloid umbellatine

Psychotria umbellata Vell. (Rubiaceae), a Brazilian coastal woody species, produces umbellatine (also known as psychollatine), an analgesic indole alkaloid. An in vitro embryogenic regeneration protocol capable of yielding alkaloid-accumulating plants was developed. Rhizogenic calli, which were obtained from stem segments derived from rooted apical cuttings, were cultured on Murashige and Skoog's (MS) medium containing either 1 mg l(-1) NAA (naphthalene acetic acid) and no kinetin, or 5 mg l(-1) NAA + 1 mg l(-1) kinetin. Calli did not accumulate umbellatine. Segments of rhizogenic callus were cultured on complete MS medium with various concentrations of kinetin and sucrose. Plant regeneration was best in the light with 0.25 mg l(-1) of kinetin and 1.5% sucrose. After 3 months of acclimatization in soil mixture, plant survival was 81%. Leaves of 10-month-old regenerated plants yielded umbellatine concentrations equivalent to those of adult forest-grown plants.     

Minimizing the central hydrophobic domain in oleosin for the constitution of artificial oil bodies

Oleosin, a unique structural protein anchoring onto the surface of seed oil bodies by its central hydrophobic domain, stabilizes these lipid-storage organelles as discrete entities. Stable artificial oil bodies have been successfully constituted with native or recombinant oleosins. In this study, recombinant sesame oleosin with 12 residues stepwise truncated from its central hydrophobic domain of 72 residues was overexpressed in Escherichia coli, was purified to homogeneity, and was used for the constitution. Artificial oil bodies constituted by truncated oleosins with the central hydrophobic domain longer than 36 residues were as stable as native sesame oil bodies, and those constituted by truncated oleosins lacking more than half of the original central hydrophobic domain inclined to coalesce upon collision or aggregation.     

Regiospecific analysis of diricinoleoylacylglycerols in castor (Ricinus communis L.) oil by electrospray ionization-mass spectrometry

HPLC fractions of diricinoleoylacylglycerols containing one non-ricinoleoyl chain from castor oil were used to identify the regiospecific location of this non-ricinoleoyl chain on the glycerol backbone using electrospray ionization-MS3 of lithium adducts. The regiospecific ions used were from the loss of alpha,beta-unsaturated fatty acid specific at the sn-2 position. The content of 1,3-diricinoleoyl-2-oleoyl-sn-glycerols (ROR) among the three stereospecific isomers, RRO, ROR and ORR, was about 91%. The contents of other 1,3-diricinoleoyl-2-acyl-glycerols among the three stereospecific isomers were as follows: 1,3-diricinoleoyl-2-linoleoyl-sn-glycerol, 95%; 1,3-diricinoleoyl-2-linolenoyl-sn-glycerol, 96%; 1,3-diricinoleoyl-2-stearoyl-sn-glycerol, 96%; 1,3-diricinoleoyl-2-palmitoyl-sn-glycerol, 78%; and 1,3-diricinoleoyl-2-lesqueroloyl-sn-glycerol, 31%. These non-hydroxyl fatty acids were mostly at the sn-2 position of triacylglycerols in castor oil. These results suggest that phospholipase A2 hydrolysis of phosphatidylcholine (PC) containing non-hydroxyl fatty acid at the sn-2 position is either blocked or partially blocked in vivo. Phospholipase A2 hydrolysis of 2-lesqueroloyl-PC is not blocked and is similar to that of 2-ricinoleoyl-PC. Transgenic inhibition of phospholipase C hydrolysis of PC might be used to block the incorporation of non-hydroxyl fatty acids into triacylglycerols, thus increasing the content of ricinoleate in seed oil.