The effect of pretreatment with different doses of GnRH to synchronize follicular wave on superstimulation of follicular growth in dairy cattle

The objective of this study was to evaluate the efficiency of gonadotropin releasing hormone (GnRH) and GnRH doses in synchronizing follicular wave emergence as a pretreatment for superovulation in cattle. Fourteen Holstein-Friesian cows 6 days from estrus were randomly assigned to receive 100 microg (n=4), 50 microg (n=5), or 25 microg (n=5) of GnRH. Superovulation was induced with injections of porcine FSH (pFSH) twice daily, decreasing the dose (total 42 AU) over 5 days beginning 2.5 days after receiving GnRH. On the 7th and 8th injections of pFSH, 750 microg of PGF(2alpha) was also given. With the exception of one cow that was given 50 microg of GnRH, ovulation was induced in all cows from the three groups and the new follicular wave emergence was observed. The total number of follicles for the 25 microg GnRH group was less than that observed for the 100 microg GnRH group (P<0.05), although there were no differences between the 100 microg, 50 microg and 25 microg GnRH groups with respect to the number of preovulatory follicles (>or=10 mm) and CL. The numbers of normal embryos were greater for the 25 microg GnRH group than the 100 or 50 microg GnRH groups (P<0.01); however, the numbers of ova/embryos did not differ significantly between the three groups. These results suggest that 25 microg of GnRH was sufficient to induce ovulation and follicular wave emergence. On day 6 of the estrous cycle, a reduction of the dose of GnRH to synchronize follicular wave emergence as a pretreatment for superstimulation promotes transferable embryos.     

Measles virus infection induces interleukin-8 release in human pulmonary epithelial cells

Measles virus (MV) infection primarily targets epithelial cells of the respiratory tract, which have the potential to synthesize a variety of cytokines. In this report, we studied the effect of MV infection on the production of interleukin (IL)-8 by the pulmonary epithelial cells. A549 cells, a lower airway epithelial cell line, produced IL-8 after MV inoculation in a dose- and time-dependent manner. The IL-8 production was little affected by UV-inactivation of MV and scarcely suppressed by cycloheximide treatment. These results indicated that MV particle binding to and/or incorporation into cells stimulated IL-8 expression in A549 cells.     

Isolation and purification of a protective protein antigen of Erysipelothrix rhusiopathiae

The rapid growth and high survival rate of Erysipelothrix rhusiopathiae was determined using a culture of the bacterium in tryptic soy broth supplemented with 0.3% Tris-hydroxymethyl aminomethane and 0.1% Tween 80 (TT-TS broth). High concentrations of 64, 66 and 43 kDa proteins, which are associated with protection against E. rhusiopathiae infection in mice, were obtained by alkaline treatment of whole cells using 0.05-1 N NaOH. The supernatant of alkaline treated cells (alkaline extract; AE) was stable at alkaline or neutral pH. However, aggregates appeared at neutral pH in the absence of sodium dodecyl sulphate (SDS). A high yield of 64, 66 and 43 kDa proteins was obtained from strain Agata (serovar 5). The proteins were eluted from gel bands following SDS-polyacrylamide gel electrophoresis (SDS-PAGE) of the AE from strain Agata and designated P64 and P43. The amounts of P64 and P43 isolated were 0.7 and 0.3 mg/16 g of wet bacteria, respectively. In a mouse protection test, 50% protective doses (PD50) of P64 and P43 were 0.58 and 0.63 microgram, respectively. Upon Western blotting of the AE, both anti-P64 and anti-P43 antibodies reacted with the 64 and 43 kDa proteins. From these results, it is suggested that P64 is the most effective protective antigen and that P43 (43 kDa protein) is a degradation product of P64. Therefore, the 64 kDa structural proteins are associated with the induction of a protective activity against E. rhusiopathiae infection in mice.     

Long-term survival of a cow with cervical ectopia cordis

This study investigated the long-term survival of a calf with cervical ectopia cordis that grew normally, became pregnant, and calved normally. The cow showed normal cardiac function and absence of peripheral circulation abnormalities. This paper documents antemortem characteristics of the affected cow.     

Isolation and characterization of macrophages from a mixed primary culture of bovine liver cells

Previously, we developed a simple and efficient method to isolate liver macrophages from a mixed primary culture of adult rat liver cells. To extend the applicability of this method, we isolated macrophages from mixed primary cultures of bovine liver cells. Macrophage cells proliferated on the cell sheet of mixed bovine liver cells after 8-16d of culture. These cells were detached by shaking of the culture flasks. Subsequent transfer and brief incubation in plastic dishes resulted in selective adhesion of macrophages. After rinses with PBS, attached macrophages were harvested. More than 10(6) cells could be harvested from the culture flask at intervals of 2-3d for more than three weeks. The isolated cells were strongly positive for bovine macrophage markers, such as CD68, CD172a and Iba-1. These cells exhibited functional properties of macrophages, including active phagocytosis of polystyrene microbeads, proliferative response to recombinant bovine granulocyte-macrophage colony-stimulating factor, upregulation of specific inflammatory cytokine genes upon stimulation with lipopolysaccharide, and formation of multinucleated giant cells. The shaking and attachment method provides a simple and efficient alternative to obtain bovine liver macrophages without requiring complex equipment or specialized technical skills.     

Arrhythmogenicity of dopamine and its effects on hemodynamics in dogs under halothane anesthesia

The arrhythmogenicity of dopamine, its effects on cardiac function, hemodynamics, and diuresis under halothane anesthesia were evaluated in dogs. The induction time of arrhythemias and the effect of arrhythmias on cardiac function, hemodynamics, and diuresis were determined after infusion of dopamine for 30-min period at increasing doses of 3, 5, 7, 10, and 15 micrograms/kg/min. The results were as follows. 1. Arrhythmia induction percentage was 28.6% at 5 micrograms/kg/mn, 42.9% at 7 micrograms/kg/min, 25% at 10 micrograms/kg/min, and 41.7% at 15 micrograms/kg/min. The induction time of arrhythemias (sec) was 459 at 5 micrograms/kg/min, 332 at 7 micrograms/kg/min, 152 at 10 micrograms/kg/min, and 279 at 15 micrograms/kg/min. No arrhythmias were present at 3 micrograms/kg/min. 2. Heart rate and myocardial oxygen consumption was increased in the arrhythmia-induced group compared to the non-arrhythmia-induced group. 3. Myocardial contractility, mean aortic pressure, mean pulmonary arterial pressure, and diuresis increased dose-dependently in the non-arrhythmia-induced group; however, these measures were increased in the arrhythmia-induced group without regard to dose.     

Application of short-time magnetic resonance examination for intervertebral disc diseases in dogs

The usefulness of magnetic resonance (MR) is already established, but it has a disadvantage of requiring a long scanning time. A short-time examination is more or less needed so as to be more practical in veterinary clinics. A protocol of the short-time MR examination was devised based on parameters determined, and validity of the protocol was assessed through the diagnosis of clinical cases with intervertebral disc diseases. With this protocol, it was possible to complete an MR examination for the spine within 15 min. The MR images and myelographic findings were correlated well in this study, suggesting the short-time protocol of MR examination can be used in the clinical diagnosis of spinal diseases.     

Effect of modification of ovary preservation solution by adding glucose on the maturation and development of pig oocytes after prolonged storage

Oocytes lose their developmental competence during prolonged storage of the ovary. In the present study, we supplemented the preservation solution for pig ovaries (phosphate buffered saline, PBS) with glucose and preserved the ovaries for 6 h at 25 C. Subsequently, we examined the glucose concentration of the follicular fluid (FF), pH of the FF, survival rate of the granulosa cells, and maturation and developmental competence of oocytes after storage. During storage, the glucose concentration of the FF (2.1 mM), pH of the FF (7.4), and survival rate of the granulosa cells (69.5%) rapidly decreased (glucose concentration: under 1.1 mM; pH: 6.8; and survival rate: 43%). On the other hand, when the preservation solution was supplemented with glucose (15 mM), the glucose concentration of the FF increased and the survival rate of the granulosa cells improved, although the pH of the FF decreased further (from 6.8 to 6.6). In addition, supplementation with glucose significantly improved the rates of oocytes at metaphase II (0 h: 65.0%; 6 h without glucose: 23.8%; and 6 h with glucose: 43.8%) and attenuated the decline in the rates of fertilization and development that resulted from prolonged storage, although there were no significant differences. In conclusion, modification of the preservation solution by the addition of glucose increased the glucose concentration of the FF and improved the rate of maturation of pig oocytes.     

Comparison of two different nutritive conditions against the changes in peripheral blood mononuclear cells of periparturient dairy cows

To clarify the relationship between cellular immune status and nutritive condition in periparturient dairy cows, feeding content, blood profiles, and immune condition were observed in cows from two dairy herds with different types of feed content. Immunological analyses such as leukocyte population and peripheral blood mononuclear cell (PBMC) mRNA of IFN-gamma, TNF-alpha, IL-4, and IL-10, quantified by real-time RT-PCR were performed. With regard to feed content during dry periods, there were six cows in the herd with insufficient non-structural carbohydrate (NFC) intake (group I) and six cows in the herd with sufficient NFC intake (group II). Significantly lower levels of blood glucose were observed in group I between weeks -12 and 16 compared with group II. Serum cholesterol level was significantly lower in group I between weeks 2 and 10 than in group II. The numbers of CD3+ and CD4+ T cells in group I were significantly lower than those in group II in weeks 6 and 14. The numbers of CD21+ B cells were significantly lower in group I than in group II in weeks -16, -12, 2, and 10. On the other hand, the CD4+/CD8+ ratio in group II was significantly higher than group I between weeks 2 and 14. The IFNgamma/IL-4 mRNA rate in group I was significantly lower than group II in week 6. We concluded that cellular immune depression occurrs after calving in dairy cows with low nutritional status in the periparturient period.     

Energy utilisation of carbohydrate, fat and protein sources in newly hatched broiler chicks

1. TME, TMEn and metabolisability (TME/gross energy) of energy-yielding foodstuffs were determined in 1, 3 and 10 d old broiler chicks, using the assay method developed by Murakami et al. (1994), in order to characterise energy utilisation during the 10 d after hatching. 2. TME, TMEn and metabolisability of dextrin and starch were low in chicks aged 1 d, and increased with age up to 10 d. Energy values of glucose and maltose at days 1 and 3 could not be determined because of the sudden death of birds soon after the feeding. TMEn and metabolisability of cereal sources were lower in d-old chicks than in those aged 3 and 10 d. At all ages, maize was better utilised than wheat and sorghum. 3. In the fat sources (coconut oil, beef tallow and safflower oil) no age dependency was observed in TME, TMEn and metabolisability. 4. Bioavailability of soyabean meal and fish meal was lower at 1 d than at 3 d and 10 d. Energy utilisation from casein was the highest among the protein sources tested and it was not age dependent. 5. It is concluded that energy utilisation of carbohydrate and protein sources during 10 d post-hatch tended to increase with age. Among the energy yielding foodstuffs fat sources seem to be better utilised, with no age dependency.