Purification and characterization of hepatic triacylglycerol lipase isolated from rainbow trout,Oncorhynchus mykiss

Trialcylglycerol (TG) lipase was isolated and partially purified from rainbow trout liver. Triacylglycerol lipase activity was assayed by measuring¹⁴C-oleic acid release from¹⁴C-triolein.¹⁴C-oleic acid release was linear for up to two hours. Optimal activity occurred at pH 7.0 and 15°C. Most of the lipase activity was recovered in the cytosolic fraction. A 27,000-fold purification was achieved after Sepharose (Bio-gel A 0.5 M, 200–400 mesh) chromatography of a resuspended 20% ammonium sulfate fraction. The molecular weight of the trout hepatic lipase as determined by size-exclusion chromatography and by SDS-polyacrylamide gel electrophoresis was 40–43 kD. Lipase-mediated hydrolysis of TG resulted in the production of diacylglycerols, monoacylglycerols, and fatty acids. Kinetic analysis indicated that V_max=0.016 nmol/h/mg protein and that K_m=0.28 mM triolein. Lipolytic activity was enhanced in the presence of cAMP/ATP-Mg²⁺. These results suggest that the liver of trout possesses a neutral TG lipase that is responsible for mobilizing stored TG and is catalytically activated by phosphorylation.A part of this work was presented at the Annual Meeting of the American Society of Zoologists, December 26–30, 1990, San Antonio, TX.     

A review of antimicrobial peptides: defensins and related cationic peptides

Cationic antimicrobial peptides are present throughout the plant and animal kingdoms and bear striking structural and functional similarities across species lines. They provide primitive, nonspecific means of combating a variety of bacteria, fungi, enveloped viruses, and protozoa. Some are also cytotoxic against host cells, including neoplastic cells. Cationic antimicrobial peptides may play various roles in inflammation and tissue repair. Antimicrobial peptides are found in epithelial tissues regularly exposed to microbial attack as well as in cells whose primary function is defense against potential pathogens. They constitute an important part of the nonoxidative antimicrobial arsenal of leukocytes. They are preformed and/or readily synthesized when the cells are stimulated by exposure to pathogens. They exert their effects directly by inserting into membranes of target cells and forming ion channels which increase membrane permeability; however, antimicrobial peptides can also act as opsonins to facilitate phagocytosis. Resistance to defensins is a virulence factor for organisms such as Salmonella sp. The study of cationic antimicrobial peptides is increasing our understanding of innate immunity, inflammation, and the pathogenesis of genetic diseases such as specific granule disease in humans. Therapeutic applications of antimicrobial peptides are currently under investigation.     

Experimental Neospora Caninum Infection in Pregnant Dairy Heifers Raises Concentrations of Pregnancy‐Associated Glycoproteins 1 and 2 in Foetal Fluids

Plasma concentrations of PAG‐1 are used for pregnancy diagnosis and as a marker of placental/foetal well‐being, while those of PAG‐2 may be an indicator of abortion risk in Neospora caninum‐infected cows. Studies have shown that N. caninum infection modifies PAG‐1 and PAG‐2 patterns in maternal blood plasma. However, no prior work has examined the effects of N. caninum infection on concentrations of PAGs in foetal fluids. In this study, PAG‐1, PAG‐2 and pH levels were determined in the amniotic and allantoic fluids of foetuses collected at 152 days of gestation from control uninfected dams and from dams experimentally infected with N. caninum on Day 110 of gestation. Foetal fluids from infected foetuses had significantly higher PAG‐2 concentrations (p = 0.026) and pH values (p = 0.02) than fluids from non‐infected foetuses. In infected foetuses, significantly higher concentrations of PAG‐1 (p < 0.001) and PAG‐2 (p < 0.001) were detected in fluid samples showing antibodies against N. caninum than those without antibodies. Moreover, pH values were significantly higher (p = 0.011) in foetal fluid samples with antibodies than in samples from non‐infected foetuses. In conclusion, this is the first report on the effect of N. caninum infection on PAG levels in foetal fluids. Our results indicate that following the experimental infection of dams with N. caninum on Day 110 of gestation, foetal fluids collected from the infected foetuses of these dams featured higher PAG‐1 and PAG‐2 levels and pH values than fluids from non‐infected controls, provided that the samples tested showed the presence of antibodies. The clinical implications of these findings are that following infection with N. caninum, most cows will experience some level of placental damage and that this injury correlates with foetal fluid PAG levels and pH.     

Reproductive biology of pond reared Nile tilapia, Oreochromis niloticus L.

The reproductive performance of female Nile tilapia, Oreochromis niloticus L., reared in ponds under tropical conditions was studied. The experiments used stock matured to the age of 2–4 months and a size of 30–50 g. Oocyte development proved a better indicator for the determination of the size at first maturation than the development of the gonado-somatic index. The Nile tilapia became ‘stunted' in the sense that early breeding took place, but the somatic growth of the stock still continued. This result indicates that the earlier breeding of Nile tilapia is not a real bottleneck for the expansion of this production sector. No relationship was found between the condition factor and the size at first maturation or the relative fecundity. This led to the conclusion that aggravation of the living conditions is not a major cause of earlier breeding of pond-reared O. niloticus.     

Effect of the Type and Number of Prostaglandin Treatments on Corpus Luteum, the Largest Follicle and Progesterone Concentration in Dairy Cows

Lactating dairy cows (n = 72) with a mature corpus luteum (CL) (diameter of > or = 17 mm) determined by ultrasonography and having a follicle with a diameter of > or = 10 mm were randomly assigned to four groups. Cows were treated with cloprostenol i.m. once or twice, or with dinoprost i.m. once or twice with an 8-h interval. The ovaries of each cow were scanned daily by transrectal ultrasonography to measure the changes in the areas of CL and the largest follicle and to determine the occurrence of ovulation. Oestrus was verified twice daily. In addition, blood sample was withdrawn from each cow daily for measuring progesterone (P4) concentrations. Significant decreases in the percentage changes relative to areas of CL and P4 concentrations or increases in the percentage changes in the area of the largest follicle on day 0 were detected in each group during the experiment. However, the type of the drug and the number of the treatments had no significant effect on those parameters. Cows ovulated with or without showing oestrus (group A) and cows exhibiting no oestrus and ovulation (group B) were also evaluated. In contrast to the mean area of the CL and the mean concentration of P4 on day 0, the mean area of the largest follicles between the two groups on day 0 differed significantly. Significant decreases in the percentage changes relative to the area of the CL and P4 concentration or increases in the percentage changes relative to the area of the largest follicle during the experiment were detected in both groups; however, there were no group differences. Treatment of dairy cows with two injections of prostaglandins (cloprostenol or dinoprost) at an 8-h interval resulted in more cows being observed in oestrus within 5 days after treatment and having significantly higher pregnancy rate than those treated with a single prostaglandin injection.     

Effect of sodium bicarbonate and sodium bentonite on digestion, solid and liquid flow, and ruminal fermentation characteristics of forage sorghum silage-based diets fed to steers

Six ruminally cannulated steers, five Holsteins and one Hereford (250 to 295 kg), were fed 84% forage sorghum silage plus 16% supplement or 50% forage sorghum silage plus concentrate and supplement diets containing either no addition (controls), 1% sodium bicarbonate (NaHCO3) or 2% sodium bentonite in a 2 X 3 factorial arrangement of treatments in a 6 X 6 Latin-square experiment with 3-wk periods. Sodium bicarbonate increased dry matter (DM) intake when concentrate was included, but neither compound affected intake of the 84% silage diet. Bentonite lowered DM, neutral detergent fiber (NDF), and acid detergent fiber (ADF) digestibilities, but NDF disappearance from nylon bags was unchanged. Ruminal pH, osmolality and L(+) and D(-) lactate were not affected by treatment. Both NaHCO3 and bentonite tended to lower ruminal NH3-N concentrations. Bentonite lowered the molar proportion of isobutyrate in ruminal fluid relative to controls, but proportions of other volatile fatty acids (VFA) and total VFA concentrations were unchanged. Neither NaHCO3 nor bentonite affected ruminal liquid or solid volumes, dilution rate constants or ruminal outflow rates. Markers overestimated volumes, but correction with measured volumes did not change interpretation of treatment effects. It was concluded that control diets had sufficiently high baseline values of pH, dilution rate and acetate proportion to preclude changes induced by either compound, especially at 1 or 2% of DM intake. An effect on palatibility through neutralization of silage acids may have been responsible for the intake response to NaHCO3.     

Evaluation of low-ash poultry meal as a protein source in canine foods

Eight mature female dogs (18.0 +/- 0.2 kg) were used in a replicated 4 x 4 Latin square experiment to determine the feeding value of low-ash poultry meal (PM) in a complete food fed to dogs. All foods contained graded concentrations of PM (10.4 to 32.5% DM), resulting in foods that were 10, 15, 20, and 25% CP. Daily DMI averaged 284 +/- 14 g/d. An increase in PM resulted in an increase in fecal moisture from 44.7 to 55.1% (linear; P < 0.01), and fecal DM output increased from 24.8 to 31.6 g/d (linear; P < 0.05). Ileal DM flow increased from 27.1 to 40.7 g/d (linear; P < 0.01). Small intestinal DM digestibility decreased from 90.4 to 86.1% (linear; P < 0.01) and total-tract DM digestibility decreased from 91.2 to 89.4% (linear; P < 0.01) as PM increased. Large intestinal DM digestibility increased from 8.4 to 21.1% with increasing PM (linear; P < 0.05). Fecal excretion of CP increased from 5.6 to 10.0 g/d (linear; P < 0.01) and ileal flow of CP increased from 6.9 to 15.6 g/d (linear; P < 0.01) as PM increased. Small intestinal CP digestibility was unaffected with treatment (P > 0.05). Large intestinal CP digestibility increased from 21.6 to 37.1% (linear; P < 0.05) with increasing PM. Total-tract CP digestibility increased from 81.0 to 86.6% (linear; P < 0.01) as PM increased. Arginine had the highest overall digestibility ranging from 88.5 to 91.3%, whereas cysteine had the lowest digestibility, ranging from 67.1 to 71.4%. These data indicate that PM is a highly digestible protein source for canine foods with inclusions of 10.4 to 32.5% of DM.     

Pancreatic exocrine secretion in steers infused postruminally with casein and cornstarch

Our objective was to evaluate the effect of postruminal protein infusion on pancreatic exocrine secretions. One Holstein, two crossbred, and five Angus steers (305 +/- 5 kg) with pancreatic pouch-duodenal reentrant cannulas and abomasal infusion catheters were used in a replicated 4 x 4 Latin square. All steers were abomasally infused with 1,050 g/d of raw cornstarch with treatments of 0, 60, 120, or 180 g/d of sodium casein suspended in water to yield 6,000 g/d of infusate daily. Steers were limit-fed (1.5 x NEm; 12 equal portions daily) a 90% corn silage, 10% supplement diet formulated to contain 12.5% CP. Periods consisted of 3 d of adaptation to infusion, 7 d of full infusion, 1 d of collection, and 7 d of rest. Pancreatic juice was collected in 30-min fractions continuously for 6 h. Total juice secreted and the pH of individual fractions were recorded, a 10% subsample was retained to form a composite sample, and remaining fluid was returned to the duodenum. Juice composite samples were stored (-30 degrees C) until analyzed for total protein and activities of alpha-amylase, trypsin, and chymotrypsin. Casein infusion linearly increased alpha-amylase concentration (182 to 271 units/mL; P < 0.02; 17.5 to 24.6 units/mg of protein; P < 0.03) and secretion rate (26,847 to 41,894 units/h; P < 0.01). Total juice secretion (155 g/h), pH of pancreatic juice (8.13), secretion rate of protein (1,536 mg/h), and concentration of protein (10.2 mg/mL) in pancreatic secretions were not affected (P > 0.05) by casein infusion. Similarly, casein infusion did not change 0.05) trypsin and chymotrypsin concentrations (1,379 and 349 units/L or 0.134 and 0.033 units/mg of protein, respectively) or secretion rates (206 and 52 units/h, respectively). Abomasal infusion of protein with starch stimulated a greater pancreatic secretion of alpha-amylase activity into the intestine than infusion of starch alone.