Background data for hematological and blood chemical examinations in juvenile beagles.

As the first step to get historical background data for physiological examinations in juvenile dogs, hematology and blood chemistry data obtained from juvenile beagle dogs (less than 3 months of age) used in the control group of toxicity studies conducted in our laboratory were summarized and compared with those obtained from adult beagle dogs (6 months of age). In the hematological examination, growth of beagle dogs was shown to be associated with increases in erythrocyte parameters and with decreases in reticulocyte and leukocyte counts. In the blood chemical examination, growth of beagle dogs was shown to be associated with increases in aspartate aminotransferase, alanine aminotransferase, and creatinine and with decreases in creatine phosphokinase, glucose, total cholesterol, and calcium. The differential leukocyte ratio showed no age relation, but the actual count showed a tendency toward decrease. Alkaline phosphatase showed a tendency to increase from 0 months of age to 3 months of age, but it decreased at 6 months of age. The present results were roughly similar to those previously reported.

Effects of restricted feeding on fetal and placental development in pregnant rabbits.

In our previous study on the effects of restricted feeding on pregnant rabbits (Matsuoka et al., 2009), animals given 20 g/day of diet on and after gestation day 6 (GD 6) showed significant changes in blood coagulation-related parameters suggesting a tendency to bleed and a decrease in serum concentration of progesterone, an important factor to maintain pregnancy, on GD 22, and a half of them showed serum progesterone concentrations less than 4.0 ng/ml which resulted in abortions on and after GD 23. In the present study, the effects of restricted feeding of 20 g/day from GD 6 to GD 22 on embryo-fetal and placental development on GD 23 as well as on blood coagulation-related parameters and serum progesterone concentrations on GD 22 were examined in pregnant rabbits. As compared with the non-restricted feeding (Not-treated, NT) group, the restricted feeding (RF) group showed lower values of platelets, fibrinogen, activated partial thromboplastin time (APTT) and antithrombin III (ATIII) and a longer prothrombin time (PT), reflecting an inhibition of blood coagulation, and a decrease in serum progesterone concentration on GD 22. Cesarean section performed on GD 23 revealed that the RF group showed a tendency towards an increase in the embryo-fetal death index and lower body weights and placental volumes compared with the NT group. Histological examination of the placenta in the RF group revealed that the labyrinth zone was thin and many glycogen-containing cells still remained in the basal zone, suggesting a delay in placental growth.

Changes in blood parameters in pregnant Japanese White rabbits.

Changes in blood parameters, especially those related to blood coagulations, were examined on gestational days (GDs) 6, 18 and 28 in Japanese White rabbits. As compared with the non-pregnant group, the following changes were observed in the pregnant group. For blood coagulation-related parameters, fibrinogen, platelets and antithrombin time III (ATIII) was significantly increased on GD 18 and/or GD 28, and activated partial thromboplastin time (APTT) was significantly prolonged on GDs 18 and 28. For the other hematological and blood chemical parameters, hemoglobin and hematocrit values were significantly decreased on GDs 18 and 28. Reticulocyte counts significantly and prominently increased on GD 18 and then decreased to significantly lower levels than those in the non-pregnant group on GD 28. Eosinophils and basophil showed significant decreases on GD 18 and/or GD 28. The serum progesterone concentration reached its highest level on GD 18. Total protein (TP), albumin, alkaline phosphatase (ALP), alanine aminotransferase (ALT), cholesterol, calcium, glucose and blood urea nitrogen were significantly decreased on GD 18 and/or GD 28. These results were almost similar to those in the pregnant New Zealand White rabbits (Mizoguchi et al., 2010).

Changes in blood parameters in New Zealand White rabbits during pregnancy.

Changes in clinical pathology parameters, particularly those related to blood coagulation, were examined throughout the gestation period in New Zealand White rabbits. As compared with the non-pregnant group, the following major changes were observed in the pregnant group. For blood coagulation-related parameters, platelets increased progressively and fibrinogen increased slightly from organogenesis, prothrombin time was significantly prolonged during organogenesis and shortened in the late fetal growth stage, activated partial thromboplastin time was significantly prolonged during the fetal growth stage, and antithrombin III increased during and after late organogenesis. Such changes in blood coagulation-related parameters during the later stages of gestation seem to be physiological responses in preparation for protecting against excessive haemorrhage or haemostasis at parturition. For the other haematological and blood chemical parameters as well as progesterone, red blood cell counts, haemoglobin and haematocrit began to decrease during organogenesis and continued to decrease thereafter. Reticulocyte counts significantly increased during organogenesis and decreased thereafter. White blood cell parameters, except for neutrophils, showed significant decreases during the fetal growth stage. Serum progesterone concentration reached its highest level early in organogenesis and decreased thereafter. Total protein, albumin, glucose, cholesterol, calcium, blood urea nitrogen and creatinine decreased significantly during the middle and/or late periods of gestation. In conclusion, the data obtained from the present study can be used as background data for effective evaluation of reproductive toxicology in rabbits, and pregnant rabbits may serve as models of pregnant women in research pertaining to clinical pathology and gestation.

Effects of restricted feeding on blood parameters in pregnant rabbits.

The effects of restricted feeding (20 g/day from gestational day (GD) 6 to 28) on pregnancy outcome and blood parameters were examined in pregnant rabbits. As compared with the group which was allowed free access to diet throughout the gestational period (NT group), the group subjected to restricted feeding (R group) showed significantly lower values in many parameters such as total protein, albumin and triglyceride on GDs 22 and 28, reflecting low nutritive conditions. In addition, there were significant changes in blood coagulation-related parameters, suggesting an imbalance between coagulation and anti-coagulation factors. Moreover, abortions occurred in about half of the animals of the R group between GDs 23 and 27. When aborted rabbits were compared with those which could maintain pregnancy under restricted feeding, total protein, albumin, platelets and antithrombin III values and especially blood progesterone concentration were significantly lower in aborted rabbits on GD 22, prior to occurrence of abortion. These results suggested that abortions due to restricted feeding might be brought about by lower nutritive conditions, an imbalance of blood coagulation-related factors and lower blood progesterone concentrations.

Age-dependent changes in the activity of serum alkaline phosphatase in laboratory beagle dogs.

In order to prepare background data for toxicity studies, serum alkaline phosphatase activity in a total of 5,242 male and female beagle dogs was surveyed for the sequence of changes in activity through aging. About 95% of the beagle dogs surveyed were 5 to 12 months of age, corresponding with the age usually employed in toxicity studies. Serum alkaline phosphatase (ALP) activity, about 460 IU/l at 5 months of age, steadily decreased and reached a level about one third of that (about 160 IU/l) at 12 months of age, and remained unchanged thereafter. The above findings were essentially the same irrespective of sex and breeding colony. The present results are useful information in the evaluation of blood chemistry data in toxicity studies.

Effects of stage and degree of restricted feeding on pregnancy outcome in rabbits.

This study's aim is to investigate the relationship between stage and degree of restricted feeding during the gestation period and occurrence of abortion, premature birth or fetal damage in rabbits. The study was composed of 5 groups of pregnant Kbl:NZW rabbits that consisted of 8 animals each. These groups were subjected to restricted feeding in the following ways: (A) control group, free access to food, (B) 60 g per day from gestational days (GD) 6 to 18 (middle period), (C) 20 g per day from GD 6 to 18, (D) 20 g per day from GD 19 to 28 (post-middle period), and (E) 20 g per day from GD 6 to 28 (middle and post-middle periods). Even though all dams in Groups A, B and C went to full term, abortion or premature birth occurred to 2/8 and 8/8 dams in Groups D and E, respectively. Fetal lethality increased in Group C, which was subjected to restricted feeding at 20 g/head/day in the middle period. Slight inhibition of fetal growth was recorded only in Group D, which was subjected to restricted feeding in the post-middle period. Restricted feeding at 20 g/head/day in the middle period induced no abortion or premature birth, but increased fetal lethality that in the middle and post-middle periods resulted in abortion or premature birth of all dams, and that in the post-middle period resulted in abortion or premature birth at low incidence and slightly inhibited fetal growth. These results demonstrated that the post-middle period is vulnerable to effects of reduced food consumption in pregnant rabbits.

Oral glucose tolerance test and determination of serum fructosamine level in beagle dogs.

The present communication deals with information regarding the practice of the oral glucose tolerance test and determination of serum fructosamine in laboratory beagles. In the oral glucose tolerance test, a 180-min level was found to be crucial following a gavage administration of 50% glucose solution at 5 mL/kg per body weight under fasting conditions. Serum fructosamine concentration as 'determined by enzymatic assay ranged between 82 and 123 micromol/L (mean of 104 micromol/L), which was about 0.285 to 0.25 times the value obtained by the chemical method described by Johnson and colleagues. Reasons for differences are ascribed to the presence of substances with reducing potential other than fructosamine in the serum.