Injection and sampling methods for drug residue study in calf muscle

Eight calves, weighing 50-150 kg, were given intramuscularly 5 ml of ampicillin (ABPC) aqueous suspensions (200 mg potency/ml) in their right and left gluteal and femoral regions. All calves were sacrificed one hour later to confirm the location of injected drug. The drug was found in a muscle layer when injected with a needle 15 mm long to the following positions, 1. the midpoint between the central position of the gluteal region (CG) and the tuber coxae (M-CTc), 2. the midpoint between CG and the tuber ossis ischii (M-CTo), 3. the central position of M. semimembranaceus in the femoral region (CF). Seven calves, weighing 130-150 kg, were given intramuscularly 5 ml of ABPC suspensions at M-CTo and CF and sacrificed one hour (4 calves) and 3 days (3 calves) later. ABPC diffused along the long axis of the muscle fibers but not to the radial direction. ABPC was detected only in the injected muscle layer even after 3 days indicating that the drug did not diffuse to the neighboring muscles. In the injected muscle layer, concentration of ABPC was remarkably different from part to part. From these results, sampling of the injected muscle for the drug residue study was proposed as follows: 1. isolate about 100 g of muscle just under the stick point marked on the skin considering the direction of drug diffusion, and 2. isolate separately about 200 g of the surrounding muscle to confirm if the sampling is appropriate.     

Cellular injury and lipid peroxidation induced by hexavalent chromium in isolated rat hepatocytes

In order to elucidate the relationship between cellular injury and lipid peroxidation induced by hexavalent chromium (CrVI), isolated rat hepatocytes treated with any one of scavengers of active oxygen species, antioxidants or antichromium agent were incubated with K2Cr2O7 as CrVI (1 mM Cr). After the incubation, the development of lipid peroxidation was determined as thiobarbituric acid (TBA)-reacting materials in total lipid extracts from the incubated hepatocytes. Cellular injury was observed as a leakage of lactate dehydrogenase (LDH) from hepatocytes into incubation medium. The contents of reduced glutathione (GSH) in hepatocytes were also assessed. Results obtained were as follows: (1) CrVI facilitated lipid peroxidation in isolated hepatocytes after 20 min of incubation. On the other hand, the cellular injury induced by CrVI was barely observed even after 60 min of incubation. (2) The CrVI-induced lipid peroxidation was inhibited by catalase and mannitol as scavengers of active oxygen species, or N,N'-diphenyl-p-phenylenediamine and alpha-tocopherol as antioxidants. However the cytotoxicity of CrVI could not be prevented by these chemicals. (3) CrVI depleted the contents of intracellular GSH and diminished the activities of glutathione reductase (GR) and glutathione-S-transferase (GST) except glutathione peroxidase. (4) The scavengers of active oxygen species and the antioxidants could not prevent the depletion of intracellular GSH induced by CrVI. (6) Ascorbic acid, antichromium agent, prevented all of the lipid peroxidation, the cellular injury and intracellular GSH depletion induced by CrVI.(ABSTRACT TRUNCATED AT 250 WORDS)     

Distribution of Cytochrome P450-side Chain Cleavage in the Theca Interna Layers of Bovine Small Antral and Cystic Follicles

Cystic follicle is anovulatory follicular structure that is caused by an endocrine imbalance. The activity of cytochrome P450‐side chain cleavage (P450scc) is essential for the initiation of steroidogenesis in the follicle. The present study was designed to compare the frequency of cells containing P450scc between healthy and atretic small antral follicles, and among several types (I, II and III, classified based on the presence of granulosa layer) of cystic follicles. Paraffin sections of healthy (2–5 mm in diameter), atretic (2–5 mm) and cystic follicles (>25 mm) were immunohistochemically stained with rabbit polyclonal antibody to bovine P450scc. The P450scc‐positive cells were counted in four different regions of the follicles from the apical to the basal side. In small antral follicles and cystic follicles, P450scc‐positive cells were localized in the theca interna layers but not granulosa layers. The P450scc‐positive cell populations decreased in the late atretic follicles compared with the early and advanced atretic follicles at all the regions of follicle. Type III cystic follicles showed significantly lower frequencies of P450scc‐positive cells than those in the types I and II cystic follicles. These results suggest that in both small and cystic follicles in cows, total loss of granulosa cells may be associated with the reduction of frequency of P450scc‐positive cells in theca interna layer.     

Intra‐uterine Infection with Babesia bovis in a 2‐day‐old Calf

Infection with Babesia bovis was diagnosed in a 2‐day‐old female calf apparently transmitted in utero. The calf was born as the second calving to a cross‐bred beef cow permanently on pasture. Diagnosis was based upon identification of B. bovis in peripheral blood smears and clinical signs which included fever, jaundice, pale mucous membranes and convulsions. Anaemia, leucocytosis, thrombocytopenia and lymphocytosis were noted at the febrile acute stage of the disease. The blood smears revealed evidence of regeneration of toxic neutrophils with a left shift, severe spherocytosis and high degree of basophilic stippling. Elevated concentration of aspartate aminotransferase, lactate dehydrogenase, and creatine kinase were also noted, and were probably the result of haemolysis, dehydration and muscle damage because of recumbancy. Elevated total bilirubin concentration following haemolysis resulted in jaundice. The neurological symptoms observed were probably caused by sludging of parasitized erythrocytes in the brain capillaries. The calf recovered following treatment with diminazene aceturate and the recovery was followed up clinically, haematologically and biochemically.     

Epidemiological studies on the expectation of life for cats computed from animal cemetery records

Based on the Chiang's method, the life table for cats was constructed from the death data of 3936 cats. They died in the Kanto area and were buried in an animal cemetery in Tokyo from June 1981 through May 1982. This life table seems to be the first one for domestic pet cats. The expectation of life for cats was 4.2 years at age 0, 5.0 years at age 1, 5.4 years at age 4, 5.3 years at age 5, 3.5 years at age 10, and 2.2 years at age 15. The maximum age at death was 22 years. From age 0 to age 5, the probability of dying for cats was higher than that for dogs, but over 6 years of age it seemed that Gompertz's equation was applicable to this life table for cats. From these results, if the probability of dying for cats at early ages decreases, the fundamental pattern of dying curve for cats seems to be a similar figure of dogs. The life table was constructed for different breeds and localities. Comparing the expectation of life at age 1 (e1) of the two populations divided by breeds or localities, there was significant difference in the e1 among different localities but not among different breeds. These facts suggest the existence of some factors which may influence the life span of cats among different localities.     

Detection of early pregnancy factor in superovulated mice

Rosette inhibition tests for the detection of early pregnancy factor (EPF) were performed on naturally ovulated and superovulated mice from day 2 of pregnancy up to 4 days after parturition. In both groups of mice, the rosette inhibition titre (RIT) increased on day 2 of pregnancy, and persisted at high levels until day 15. Thereafter, the RITs of both groups of mice decreased to the non-pregnancy range. No significant differences of the mean RITs between these two groups were observed during the high RIT period. These results showed that the superovulatory treatment did not cause any changes or interference in the detection of EPF. In order to investigate the initial time of appearance of EPF in the maternal circulation in relation to the stage of fertilization, measurement of RIT and examination of the fertilization stage were carried out on superovulated mice 1 day after mating. The mean RIT of mice with pronucleus stage ova was significantly (p less than 0.01) higher than that of mice with sperm-penetrated ova. EPF was considered to appear in the maternal peripheral blood at the pronucleus stage.     

Type-I hypersensitivity as a component of eosinophilic myositis (muscular sarcocystosis) in cattle

Eight bovine hearts with lesions of eosinophilic myositis (EM) and 2 bovine hearts without EM lesions were collected at slaughter. Blood samples from these 10 hearts, and the heart of a newborn calf also were collected. Histologically, Sarcocystis cruzi was identified in the 8 hearts with EM lesions and the 2 hearts without EM lesions, but not in the heart of the newborn calf. Serum was harvested from the 10 blood samples and was used in homologous, modified, passive cutaneous anaphylaxis tests. Antigen was prepared from S cruzi bradyzoites isolated from the 2 hearts without EM lesions. Serum samples from the 8 cattle with EM lesions reacted positively to S cruzi antigen. When heat-inactivated IgE in serum (56 C for 4 hours) was used, all passive cutaneous anaphylaxis responses were considered negative. Using ELISA, serum IgE concentrations from the 10 cattle with and without EM lesions were 2.2 to 9 U/ml. As determined by radial immunodiffusion, IgM concentrations were 80 to 215 mg/dl. Immunoglobulin G concentrations were 420 to 2,050 mg/dl, but most were less than or equal to 1,700 mg/dl. Immunoglobulin A concentrations were 0 to 62 mg/dl; 1 steer with EM lesions had 0 mg/dl. Double-gel immunodiffusion confirmed the presence of Sarcocystis-specific precipitating antibodies. Sera from the 10 cattle with and without EM lesions formed at least 1 precipitin band.     

Effects of abiotic variables on the response of carnation cultivars to Fusarium oxysporum f.sp. dianthi

The effects of abiotic variables on the response of carnation cultivars to Fusarium oxysporum f.sp. dianthi ( F.o. dianthi  ) were examined in experiments conducted under semi-controlled environments. The abiotic variables examined were solar radiation intensity, temperature and growth substrate. Temperature was not controlled, but differed markedly among experiments, thus, its effect was not determined quantitatively. Disease incidence and disease severity varied significantly among the experiments (due mainly to differences in temperature), among the solar radiation treatments and among the cultivars tested. The three-way interaction term (i.e. cultivar × shade treatment × experiment) was highly significant ( P  < 0.001) when both disease incidence and disease severity were considered, indicating that no single variable was predominant in determining disease intensity. The effects of the growth substrate on disease progress was examined in plants grown in tuff or in tuff mixed with peat (1 : 1 and 1 : 3) substrates. The growth substrate had a potent effect on disease development in the less susceptible cultivars. Severe epidemics developed in all cultivars when they were grown in the tuff/peat mixture, although some were resistant when grown in tuff alone. These results led to the conclusion that the carnation response to F.o. dianthi is substantially influenced by the environmental conditions of the test.     

The effects of stubble burning and tillage on soil carbon sequestration and crop productivity in southeastern Australia

Abstract. In Australia, stubble burning and tillage are two of the major processes responsible for the decline of soil organic carbon concentration in cropped soils, and the resulting soil degradation. However, the relative importance of these two practices in influencing the soil organic carbon concentration and the long-term impact on soil quality and productivity are not clear. The effects of stubble burning as practised by farmers in southeastern Australia were evaluated in two field trials, one of 19 years duration, the other of 5 years. Conventional tillage (three tillage passes) led to greater loss of soil organic carbon than stubble burning. Loss of total soil organic carbon attributed to stubble burning in the 0–10 cm layer was estimated to be 1.75 t C ha⁻¹ over the period of the 19-year trial, equivalent to 29% of that lost due to tillage. In the 5-year trial, no change in soil organic carbon due to stubble burning was detectable. Changes in soil quality associated with stubble burning detected in the longer trial included a reduction in macro-aggregate stability, and increases in pH and exchangeable K⁺. Only the latter two were detected in the shorter trial. A higher mean wheat yield (average 0.15 t ha⁻¹) following stubble burning was observed in the 19-year trial but not in the 5-year trial. Research to monitor the longer term effects of stubble burning is needed, and to identify conditions where loss of soil organic carbon is minimized.