Use of canine red blood cell with high concentrations of potassium, reduced glutathione, and free amino acid as host cells for in vitro cultivation of Babesia gibsoni

OBJECTIVE: To determine the usefulness of canine RBC with high concentrations of potassium, reduced glutathione (GSH), and amino acid(i.e., HK cells) for in vitro cultivation of Babesia gibsoni. ANIMALS: RBC were obtained from 3 dogs that had inherited HK cells and from 3 genetically unaffected dogs that, therefore, had RBC with lower potassium (LK) concentrations (i.e., LK cells). PROCEDURES: First, B. gibsoni were cultivated using HK or LK cells in alpha-modification of Eagle medium, consisting of Earle salts with glutamine and without ribosides, deoxyribosides, and sodium bicarbonate under a humidified atmosphere containing 5% CO2 at 37 C. Second, parasites were cultivated with LK- or HK-cell lysates. Finally, HK cells were separated into 3 fractions (bottom, middle, top layers) by density gradient centrifugation, and B. gibsoni were cultivated with each of the HK-cell fractions. In addition, the concentrations of free amino acids and reduced glutathione (GSH) in each HK-cell fraction were measured. RESULTS: B. gibsoni preferentially multiplied in HK-cell cultures rather than in LK-cell cultures. Furthermore, the addition of HK-cell lysate to the culture medium resulted in enhanced multiplication of the parasites. Higher multiplication of the parasites was observed in HK cells from the top layer, compared with HK cells from the middle and bottom layers. The HK cells from the top layer had higher concentrations of glutamate, aspartate, and GSH, compared with HK cells from the middle and bottom layer. CONCLUSIONS: Canine HK cells are useful host cells for in vitro cultivation of B. gibsoni, and the high concentrations of glutamate, aspartate, and GSH may result in enhancement of multiplication of the parasites in HK cells.     

Methane output and diet digestibility in response to feeding dairy cows crude linseed, extruded linseed, or linseed oil

This experiment studied the effect of 3 forms of presentation of linseed fatty acids (FA) on methane output using the sulfur hexafluoride tracer technique, total tract digestibility, and performance of dairy cows. Eight multiparous lactating Holstein cows (initial milk yield 23.4 +/- 2.2 kg/d) were assigned to 4 dietary treatments in a replicated 4 x 4 Latin square design: a control diet (C) consisting of corn silage (59%), grass hay (6%), and concentrate (35%) and the same diet with crude linseed (CLS), extruded linseed (ELS), or linseed oil (LSO) at the same FA level (5.7% of dietary DM). Each experimental period lasted 4 wk. All the forms of linseed FA significantly decreased daily CH(4) emissions (P < 0.001) but to different extents (-12% with CLS, -38% with ELS, -64% with LSO) compared with C. The same ranking among diets was observed for CH(4) output expressed as a percentage of energy intake (P < 0.001) or in grams per kilogram of OM intake (P < 0.001). Methane production per unit of digested NDF was similar for C, CLS, and ELS but was less for LSO (138 vs. 68 g/kg of digested NDF, respectively; P < 0.001). Measured as grams per kilogram of milk or fat-corrected milk yield, methane emission was similar for C and CLS and was less for ELS and LSO (P < 0.001), LSO being less than ELS (P < 0.01). Total tract NDF digestibility was significantly less (P < 0.001) for the 3 supplemented diets than for C (-6.8% on average; P < 0.001). Starch digestibility was similar for all diets (mean 93.5%). Compared with C, DMI was not modified with CLS (P > 0.05) but was decreased with ELS and LSO (-3.1 and -5.1 kg/d, respectively; P < 0.001). Milk yield and milk fat content were similar for LSO and ELS but less than for C and CLS (19.9 vs. 22.3 kg/d and 33.8 vs. 43.2 g/kg, on average, respectively; P < 0.01 and P < 0.001). Linseed FA offer a promising dietary means to depress ruminal methanogenesis. The form of presentation of linseed FA greatly influences methane output from dairy cows. The negative effects of linseed on milk production will need to be overcome if it is to be considered as a methane mitigation agent. Optimal conditions for the utilization of linseed FA in ruminant diets need to be determined before recommending its use for the dairy industry.     

Pharmacokinetics and bioavailability of cefquinome in healthy piglets

A study on bioavailability and pharmacokinetics of cefquinome in piglets was conducted after intravenous (i.v.) and intramuscular (i.m.) administrations of 2.0 mg/kg of body weight, respectively. Plasma concentrations were measured by high‐performance liquid chromatography assay with UV detector at 268‐nm wavelength. Plasma concentration–time data after i.v. administration were best fit by a two‐compartment model. The pharmacokinetic values were distribution half‐life 0.27 ± 0.21 h, elimination half‐life 1.85 ± 1.11 h, total body clearance 0.26 ± 0.08 L/kg·h, area under curve 8.07 ± 1.91 μg·h/mL and volume of distribution at steady state 0.46 ± 0.10 L/kg. Plasma concentration–time data after i.m. administration were also best fit by a two‐compartment model. The pharmacokinetic parameters were distribution half‐life 0.88 ± 0.42 h, elimination half‐life 4.36 ± 2.35 h, peak concentration 4.01 ± 0.57 μg/mL and bioavailability 95.13 ± 9.93%.     

Effects of dietary oregano and garlic essential oils on carcass characteristics,meat composition,colour, pH and sensory quality of broiler meat

1. An experiment was conducted to determine the individual and combined effects of two essential oils, oregano and garlic, on carcass characteristics, meat composition, colour, pH and sensory quality of broiler meat.

2. The diets were supplemented with no essential oil (control), oregano essential oil or garlic essential oil at 300 mg/kg and oregano essential oil at 150 mg/kg + garlic essential oil at 150 mg/kg.

3. Dietary oregano and garlic oil supplementation did not affect carcass yields, the relative weight of carcass parts, breast and thigh meat composition, pH or b* value of breast meat. Oregano + garlic oil supplementation significantly decreased the L* value. The a* value of breast meat in birds given a diet supplemented with oregano oil was lower than that in birds given a diet supplemented with garlic oil and oregano oil + garlic oil. The essential oil addition had no positive effect on the oxidative stability. There was no difference between the treatments in breast appearance.

4. The juiciness, flavour, oxidised flavour and acceptability of breast meat samples were affected by treatments.     

The muscular dystrophic chicken is hypernatremic

1. The E3 ubiquitin protein ligase 1 (WWP1) gene, the mutation of which causes muscular dystrophy in chickens, is expressed not only in the pectoral muscle, but also in a number of tissues such as the kidney. Therefore, this study examined some parameters related to kidney function in muscular dystrophic (MD) chickens.

2. Plasma osmolality, Na⁺ and K⁺ concentrations, aldosterone levels, and the expression of aquaporin (AQP) 2, AQP3, and α subunits of the amiloride-sensitive epithelial sodium channel (αENaC) were analysed in the kidneys of 5-week-old MD chickens and White Leghorn (WL) chickens under physiological conditions or after one day of water deprivation.

3. Plasma osmolality, Na⁺ concentrations, and plasma aldosterone levels were significantly higher in MD chickens than in WL chickens. αENaC mRNA expression levels were lower in MD chickens than in WL chickens. AQP2 and AQP3 mRNA expression levels were similar in the two strains of chickens.

4. Plasma osmolality correlated with aldosterone levels and AQP2 and αENaC mRNA levels in WL chickens. In MD chickens, plasma osmolality correlated with AQP2 mRNA levels, but not with plasma aldosterone or αENaC mRNA levels.

5. These results suggest that neither water reabsorption nor the expression of AQP2 and AQP3 is impaired in MD chickens and that a WWP1 gene mutation may or may not directly induce an abnormality in Na⁺-reabsorption in the kidneys of MD chickens, potentially through αENaC.     

Estimation of genetic correlations between racing times recorded at different racing distances by restricted maximum likelihood in Thoroughbred racehorses

SUMMARY: Genetic correlations between racing times on track type (turf and dirt), and at racing distances on turf (1200 m, 1400 m, 1600 m, 1800 m, and/or 2000 m) and dirt (1000 m, 1200 m, 1400 m, 1600 m, 1700 m, and/or 1800 m) tracks, were estimated in Thoroughbred horses. (Co)variance components were estimated using multiple-trait derivative-free restricted maximum likelihood (MTDFREML). The data used were collected by the Japan Racing Association from 1992 to 1993. The generation 2 pedigree information was preferable for (co)variance estimates. The genetic correlations between racing times on turf and dirt tracks ranged from 0.69 to 0.31 (average 0.51). The genetic correlations between racing distances ranged from 0.68 to 1.00 (average 0.85) and from 0.53 to 1.00 (average 0.88) on turf and dirt tracks, respectively. These results suggest that the racing time per 100 m can be used for horse genetic evaluation within one track type. ZUSAMMENFASSUNG: Sch?tzung genetischer Korrelationen zwischen Rennzeiten von Vollblüternüber verschiednen Distanzen mittels restringierter Genetische Korrelationen zwischen Rennzeiten auf Rasen- und Erdbahnen, Renndistanzen auf Rasen- (1200, 1400, 1600, 1800 und 2000 m) und Erdbahnen (1000, 1200, 1400, 1600, 1700 und 2000 m) wurden für Vollblüter gesch?tzt. (Co)Varianzkomponenten wurden mittels Mehr-Merkmal Ableitungsfreier Restringierter Maximum Likelihood (MTDFREML) gesch?tzt. Die Unterlagen wurden von der Japanischen Renn Vereinigung 1992 und 1993 gesammelt. Generation 2 Abstammungsinformation war für die Co-Varianzsch?tzung günstig. Genetische Korrelationen zwischen Rennzeiten auf Rasen und auf Erdbahnen waren zwischen 0.69 und 0.31 (Durchschnitt 0.51), jene zwischen Distanzen zwischen 0.68 und 1.00 (Durchschnitt 0.85) und zwischen 0.53 und 1.00 (Durchschnitt 0.88) auf Rasen und Erdbahnen. Die Ergebnisse zeigen, da? Rennzeit per 100 m zur Bewertung der Pferde geeignet ist.     

Genetic typing of classical swine fever virus

Three regions of the classical swine fever virus (CSFV) genome that have been widely sequenced were compared with respect to their ability to discriminate between isolates and to segregate viruses into genetic groups. Sequence data-sets were assembled for 55 CSFVs comprising 150 nucleotides of the 5' non-translated region, 190 nucleotides of the E2 envelope glycoprotein gene and 409 nucleotides of the NS5B polymerase gene. Phylogenetic analysis of each data-set revealed similar groups and subgroups. For closely related viruses, the more variable or larger data-sets gave better discrimination, and the most reliable classification was obtained with sequence data from the NS5B region. No evidence was found for intertypic recombination between CSFVs. A larger data-set was also analysed comprising 190 nucleotides of E2 sequence from 100 CSFVs from different parts of the world, in order to assess the extent and global distribution of CSFV diversity. Additional groups of CSFV are evident from Asia and the nomenclature of Lowings et al. (1996) [Lowings, P., Ibata, G., Needham, J., Paton, D., 1996. J. Gen. Virol. 77, 1311-1321] needs to be updated to accommodate these. A tentative assignment, adapting rather than overturning the previous nomenclature divides CSF viruses into three groups with three or four subgroups: 1.1, 1.2, 1.3; 2.1, 2.2, 2.3; 3.1, 3.2, 3.3, 3.4. The expanding data-base of CSFV sequences should improve the prospects of disease tracing in the future, and provide a basis for a standardised approach to ensure that results from different laboratories are comparable.