Isolation of new pregnancy-associated glycoproteins from water buffalo (Bubalus bubalis) placenta by Vicia villosa affinity chromatography

The present study describes the isolation and characterization of new pregnancy-associated glycoprotein molecules (PAG) from midpregnancy and late-pregnancy placentas in the water buffalo (Bubalus bubalis). After extraction, the homogenates are subjected to acid and ammonium sulfate precipitations followed by DEAE chromatography. Subsequently, the water buffalo PAG (wbPAG) from these solutions are enriched by Vicia villosa agarose (VVA) affinity chromatography. As determined by western blotting with anti-PAG sera, the apparent molecular masses of the immunoreactive bands from the VVA peaks range from 59.5 to 75.8 kDa and from 57.8 to 73.3 kDa in the midpregnancy and late-pregnancy placentas, respectively. Amino-terminal microsequencing of the immunoreactive proteins has allowed the identification of three distinct wbPAG sequences, which have been deposited in the SwissProt database: RGSXLTIHPLRNIRDFFYVG (acc. no. P85048), RGSXLTILPLRNIID (acc. no. P85049), and RGSXLTHLPLRNI (acc. no. P85050). Their comparison to previously identified proteins has shown that two of them are new because they have not been described before. Our results confirm the suitability of VVA chromatography for the enrichment of the multiple PAG molecules expressed in buffalo placenta.     

The Effects of Feed Form on Consumption Time and Glucose and Insulin Response to a Concentrate Meal in Equine

This study tested the hypothesis that feeding an identically formulated, low sugar and starch concentrate in three forms (5-mm extruded [E], 4-mm pellet [P], and 19-mm oval [O]) would affect consumption rate and glucose or insulin responses, or both. Horses received 1.8 kg treatment feed in a randomized, crossover design, with samples taken every 30 minutes for 6 hours for blood glucose and insulin response. Pearson's correlation compared consumption time, insulin and glucose peak, and time to peak insulin and glucose. The pellet (P) elicited a lower (P = .01) glucose concentration at 2.5 hours than O. The pellet also elicited a lower (P = .03) insulin concentration at 5.5 hours than E and O. There were no differences (P > .05) in area under the curve (AUC) insulin, peak insulin, and time to peak insulin for the three treatments. Average insulin concentration was lower (P = .01) for P versus O. There were no differences (P > .05) in average insulin between P and E, nor between O and E. There were no differences (P > .05) in AUC and peak glucose concentration. Time to peak glucose was longer (P = .04) for P versus E. Average glucose concentration was lower (P = .02) for P versus O. Consumption time was longer (P = .03) for O versus P. There was a positive correlation between consumption time and time to peak insulin (r = 0.46, P = .029). Further research on feeding practices, feed forms, and consumption times that affect glycemic response is necessary.     

Effects of cereals and/or protein supplement extrusion on diet utilisation and performance of intensively reared cattle

The effect of extruding the cereal and/or the protein supplement of a compound feed on its nutritive value and on the performance of intensively reared male calves was studied. The compound feed was formulated with 0.65 of a cereal blend (60:40 maize:barley), 0.25 of a protein blend (1/3:1/3:1/3 raw soybeans:peas:lupins), and 0.08 of urea to contain 0.17 of crude protein. It was tested without extruding (NE) and with the cereal blend (CE), the protein blend (PE) or both (CPE) extruded. Another non-extruded compound feed with mainly soybean meal as the protein supplement (NE-SBM) also was studied. The five experimental compound feeds together with barley straw were offered ad libitum to 50 male Friesian calves (112 kg initial live weight) for 13 weeks, recording individual feed intake and live weight gain. Crude protein (CP) solubility and in vitro and in situ CP degradation of experimental compounds, and in vitro gas production kinetics, in vivo digestibility and urinary allantoin excretion of diets were determined. Cereal extrusion promoted a greater gas production at all incubation times (p < 0.001), associated to the increase in starch gelatinisation. PE and NE-SBM compounds showed lower in vitro (p < 0.01) and in situ (p < 0.001) CP degradability than the other feeds. Ingredient extrusion did not affect apparent DM, OM and CP digestibilities, but diet CPE showed a lower NDF digestibility (p < 0.05) than the others (0.293 vs. 0.420, 0.387, 0.390 and 0.407 with CPE vs. NE, CE, PE and NE-SBM, respectively). The apparent EE digestibility increased (p = 0.053) with the inclusion of extruded ingredients, (0.799, 0.749 and 0.794 vs. 0.719 and 0.702 with CE, PE and CPE vs. NE and NE-SBM, respectively). Daily allantoin excretion was lower (p < 0.01) in calves receiving the CPE diet than in those receiving the NE, PE and NE-SBM diets. Treatments did not affect the average daily gains (1.61 ± 0.022 kg d^− 1), however calves on the CPE diet showed a lower (p < 0.01) concentrate and total conversion ratios than those fed NE, PE and NE-SBM (2.60 vs. 2.79, 2.85 and 2.98 kg concentrate DM kg^− 1 daily gain and 2.93 vs. 3.09, 3.13 and 3.26 kg total DM kg^− 1 daily gain, respectively). The CE diet resulted in numerically higher (p > 0.05) concentrate and total DM conversion ratios (2.72 and 3.02) than CPE and lower (p < 0.05) than NE-SBM. Improvement in feed conversion ratio after extrusion would be related to a better starch utilisation and not to changes in microbial or dietary protein flow to the duodenum, although a better utilisation of fat energy cannot be disregarded.     

Mermithid nematode infection in a colony of blue-winged grasshoppers (Tropidacris collaris)

A die-off occurred in a captive colony of blue-winged grasshoppers (Tropidacris collaris) at the Toronto Zoo. One fourth of the colony died within a year due to infection with worms initially mistaken for nematomorphs but later identified as nematodes belonging to the Mermithidae, genus Mermis. Mortality persisted and the grasshopper population dwindled over the following years. Mermithid larvae developed in the hemocoel of the insects until they eventually emerged from a hollowed-out exoskeleton. Circumstantial evidence suggests that the parasites were introduced with raspberry browse that was grown on site and contaminated with mermithid eggs.     

Differential response of cull cow muscles to the hypertrophic actions of ractopamine-hydrogen chloride

Ractopamine-HCl (RAC) is a beta-adrenergic agonist with variable effects on cattle performance and carcass variables. Cull cows fed RAC (200 mg . head(-1) . d(-1)) demonstrate an increased size of type I and II muscle fibers that does not translate into a larger ribeye area. The objective of this study was to examine the dose-dependent effects of RAC on cull cow muscle morphometrics. Eighty-eight cull beef cows representing 2 breed types (n = 44 each) were fed 0, 100, 200, and 300 mg . head(-1) . d(-1) of RAC for the last 28 d of a 54-d feeding period. On d 54, cows were slaughtered, and samples of the LM and semimembranosus muscle (SM) from 16 randomly selected carcasses (n = 4 per treatment) were taken for measurement of beta (2)-adrenergic receptors and type I, IIA, and IIX myosin heavy chain (MyHC) gene expression. Twenty-four hours postmortem, LM, SM, infraspinatus (INF), and vastus lateralis samples from 40 randomly selected carcasses (n = 10 per treatment) were obtained and frozen for immunohistochemical analysis. Muscle fiber cross-sectional area and diameter, MyHC isoform expression, and fiber-associated nuclei numbers were measured. Ractopamine dosage exhibited differential effects on muscle morphometrics and MyHC gene expression. Muscle fiber cross-sectional area and diameter were increased (P < 0.05) by RAC in INF type I and IIA fibers and SM type IIA fibers. Ractopamine increased (P < 0.05) MyHC type IIX mRNA and tended to increase (P < 0.10) beta(2)-adrenergic receptors in the SM; a change in mRNA abundance was not detected for either gene in the LM. Treatment with RAC decreased (P < 0.05) fiber-associated nuclei numbers in the INF, vastus lateralis, and LM but did not affect (P > 0.05) MyHC or beta-adrenergic receptor expression. These results indicate that cull cow feeding programs may consider supplementing RAC as a means of adding value to cuts within the chuck, such as the INF.     

Acute lead intoxication in a pregnant mare

Lead (Pb) intoxication in horses is usually a chronic phenomenon with clinical signs associated with central nervous dysfunction. This report gives details of a case of acute Pb intoxication in a 9-year-old American Saddlebred mare with severe, progressive and ultimately fatal neurological deterioration. During the 4 days of hospitalisation, clinical signs progressed from intermittent headshaking and depression to severe, continuous, uncontrollable manic behaviour. At autopsy, three grey-coloured, hard metal particles were present in the gastrointestinal tract and subsequently found to contain 2614 ppm Pb. Lead concentrations in the brain, liver, stomach and kidney were 29, 4, 6 and 2 ppm wet weight, respectively.     

Structural heterogeneity of yolk spheres in hierarchical follicles from hen ovary: a histochemical study

1. Four types of yolk spheres with variable structure, chemical composition and frequency of occurrence in yolk plasma of hierarchical follicles (F(4), F(3), F(2) and F(1) with diameters of 10.0, 15.5, 20.0 and 35.0 mm, respectively) of the hen ovary were identified using histochemical methods for localising lipids, carbohydrates and proteins. 2. Yolk spheres of the first type (YS(1)) had a phospholipoprotein membrane surrounding fluid matrix which stained lightly for phospholipids, proteins and acidic mucopolysaccharides. Two types of droplets were observed in the matrix of YS(1). Spheres of the second type (YS(2)) had a lipoprotein- and acidic mucopolysaccharide-rich peripheral region and a single large droplet in its fluid matrix. Droplets of YS(2), unlike YS(1), showed three regions and metachromatic staining with ninhydrin-Schiff reagent. The third type of sphere (YS(3)) had a homogeneous matrix staining for proteins, neutral lipids and florescent yellow with alcian blue and differentially with ninhydrin-Schiff reagent; it was bounded by a phospholipids- and acidic mucopolysaccharide-containing thick peripheral region. Its fluid matrix also showed toluidine-blue-positive, densely packed granules and small droplets. The fourth type (YS(4)) was seen only in bromophenol blue and Nile blue preparations, revealing the presence of proteins and neural lipids in their matrix and peripheral regions. 3. Quantitative data on the relative abundance of yolk spheres in F(4) to F(1) follicles revealed more YS(3) (51.1 to 64.7%) than YS(1) (16.2 to 28.3%) and YS(2) (19.1 to 23.2%). The percentage of YS(1) increased and that of YS(3) decreased as follicle size increased.     

Intestinal metabolism of weaned piglets fed a typical United States or European diet with or without supplementation of tributyrin and lactitol

The aim of the study was to investigate the effects of supplementation of a microencapsulated blend of tributyrin and lactitol (TL) to a standard European (EU) diet without antibiotic growth promoters on intestinal metabolism and mucosa development of weaned piglets and to compare it with a standard US diet containing animal proteins, zinc oxide, copper sulfate, and carbadox. Ninety piglets weaned at 21 d were divided into 3 dietary groups consisting of 5 replicates each: 1) US diet supplemented with 55 mg/kg of carbadox, and 2.5% each of plasma proteins and spray-dried blood cells in the first phase, 3,055 mg/kg of Zn in the first and second phases, and 180 mg/kg of Cu in the third phase; 2) EU diet based on vegetable proteins and no antibiotics; and 3) the same EU diet supplemented with 3,000 mg/kg of microencapsulated TL. The study was divided into 3 phases: 0 to 7, 8 to 21, and 22 to 35 d. On d 7, 21, and 35, animals were weighed, and feed consumption and efficiency were determined. On d 14 and 35, one pig per pen was killed, and the intestinal contents and mucosa from the proximal, middle, distal jejunum and the ileum were sampled. Intestinal wall sections were fixed for histological analysis, and intestinal content was used for VFA, ammonia, and polyamine analysis. Throughout the study (d 0 to 35), the US diet had greater ADG and ADFI than the EU diet (P < 0.05). The EU diet supplemented with TL tended to have 11% greater ADG (P = 0.17). Feeding the EU diet caused a reduction in proximal and middle jejunum villi length by 10% (P < 0.05) and an increase in crypt size in proximal jejunum (P < 0.05) compared with the US diet, probably due to an increased rate of cell loss and crypt cell production. The TL supplementation resulted in longer villi along the jejunum and less deep crypts in the proximal jejunum (+15.9 and -8.9%, respectively; P < 0.05) than the unsupplemented EU diet. The TL diet increased the concentrations of cadaverine and putrescine in the small intestine (P < 0.05) and seemed to increase cadaverine, histamine, putrescine, and spermine in the large intestine by 1.5- to 10-fold compared with the US or EU diet. In conclusion, although the US diet had a greater effect on growth performance and mucosal trophic status than the EU diets, the supplementation with slowly released TL seemed to be an effective tool to partially overcome the adverse effects of vegetable protein diets.