Study of Kyasanur forest disease viremia, antibody kinetics, and virus infection in target organs of Macaca radiata.

The present manuscript deals with experimental infections of bonnet macaques (Macaca radiata) to study disease progression for better insights into the Kyasanur Forest Disease (KFD) pathogenesis and transmission. Experimentally, 10 monkeys were inoculated with KFD virus (KFDV) (high or low dose) and were regularly monitored and sampled for various body fluids and tissues at preset time points. We found that only 2 out of the 10 animals showed marked clinical signs becoming moribund, both in the low dose group, even though viremia, virus shedding in the secretions and excretions were evident in all inoculated monkeys. Anti-KFDV immunoglobulin (Ig)M antibody response was observed around a week after inoculation and anti-KFDV IgG antibody response after two weeks. Anaemia, leucopenia, thrombocytopenia, monocytosis, increase in average clotting time, and reduction in the serum protein levels were evident. The virus could be re-isolated from the skin during the viremic period. The persistence of viral RNA in the gastrointestinal tract and lymph nodes was seen up to 53 and 81 days respectively. Neuro-invasion was observed only in moribund macaques. Re-challenge with the virus after 21 days of initial inoculation in a monkey did not result in virus shedding or immune response boosting.

Assessment of ovarian follicular dynamics and folliculogenesis associated endocrine profiles following gonadotropin stimulation in the bonnet monkey.

We evaluated ovarian follicular dynamics in bonnet monkeys by employing trans-abdominal ultrasonography. Following the administration of human follicle stimulating hormone (hFSH) and/or human menopausal gonadotropin (hMG), multiple follicular development was assessed and their numbers, size and growth profiles were monitored. The ultrasonograms showed that the follicular antrum appeared distinctly anechoic with well-defined hyperechoic borders. Depending on the type, quantity (12.5-25IU), and duration (6-9days) of hormones administered, the number of developing follicles was 2-12 per ovary with their lowest diameter being 2mm. With continued hormone administration, their numbers and diameters increased; which were more pronounced in animals administered with hFSH than with hMG, with follicles of 6-8mm. Interestingly, human chorionic gonadotropin (hCG) injection (2000-3000IU), when follicles acquiring >6-8mm sizes, induced the maximum expansion of antral follicles with sizes reaching up to 14mm. On days 3-5 post-hCG, the ultrasonograms showed loosely demarcated multiple hypoechoic structures and well-demarcated hyperechoic structures with anechoic/hypoechoic cores corresponding to unruptured luteinized follicles and corpora lutea, respectively. On day 4 post-hCG, there was a substantial reduction in the number of antral follicles. In stimulated animals, follicular growth, ovulation, and formation of luteal structures were accompanied by corresponding physiological changes in the serum estradiol and progesterone profiles. These findings, for the first time, showed that ultrasonographic imaging approach is useful for precise monitoring of temporal changes in follicular developmental dynamics and to time the hCG induced ovulation in the bonnet monkey.

Hematological and serum biochemical indices in healthy bonnet macaques (Macaca radiata).

BACKGROUND: Blood reference values for bonnet macaques (Macaca radiata) are limited. The goal of this study was to determine reference ranges for hematological and serum biochemical indices in healthy, socially housed bonnet macaques for males and females over a range of ages. METHODS: Blood hematological and serum biochemical values were obtained from 50 healthy bonnet macaques of both sexes and aged 10-234 months. RESULTS: Age and sex differences were present in a number of measures. Globulins, total protein, and creatinine (CREAT) values were highest among older subjects, while alkaline phophatase, albumin, and phosphorus values were higher in juveniles. Sex differences were present in concentrations of red blood cells and CREAT, with higher values in males. CONCLUSION: The blood parameter data reported here as age-specific reference values for laboratory-housed, healthy bonnet macaques may be used to inform clinical care and laboratory primate research.

Effects of ketamine and thiopentone anesthesia on serum lipid parameters in adult bonnet monkeys (Macaca radiata).

We investigated the effect of anesthesia on serum lipid parameters in adult bonnet macaques (Macaca radiata). We treated 10 animals with ketamine hydrochloride (15 mg/kg intramuscularly) and, on the next day, thiopentone sodium (25 mg/kg intravenously). Blood samples were obtained before and after anesthetic treatment. Serum cholesterol, triglycerides, very low-density lipoprotein (VLDL), and high-density lipoprotein (HDL) were measured by autoanalyzer. Ketamine anesthesia significantly decreased serum cholesterol and HDL levels, whereas thiopentone significantly reduced triglycerides and VLDL and increased HDL values. Although the effects of ketamine hydrochloride and thiopentone sodium on serum biochemical values have been reported, no literature addressing the effect of anesthesia on lipid parameters in bonnet macaques is available. These findings will be useful in designing experiments assessing pathologic and toxicologic changes in serum lipid parameters and interpreting data obtained from adult bonnet monkeys.

Hematologic and serum biochemical values in aged female bonnet macaques (Macaca radiata) anesthetized with ketamine hydrochloride.

We investigated the effect of ketamine hydrochloride anesthesia on hematologic and serum biochemical values in 10 aged female bonnet macaques (Macaca radiata) before and 120 min after intramuscular administration of ketamine hydrochloride (15 mg/kg body weight). Ketamine anesthesia caused significant reduction in the total leukocyte count, lymphocyte count, red blood cell count, hemoglobin concentration, packed cell volume, and serum concentrations of glucose, total protein, alkaline phosphatase, calcium, sodium, and potassium. Although the effects of ketamine hydrochloride on hematologic and serum biochemical values have been reported for most of the nonhuman primates, no literature on bonnet macaques is available. These findings will be useful in designing experiments assessing pathologic and toxicologic changes in blood and serum parameters and interpreting data obtained from aged bonnet monkeys.

Normal hematological indices, blood chemistry and histology and ultrastructure of pancreatic islets in the wild Indian bonnet monkeys (Macaca radiata radiata).

The present study is aimed at determining some haematological and biochemical parameters in the wild Indian bonnet monkeys as also the microscopic and ultrastructural characteristics of their pancreatic islets. Adult wild Indian bonnet monkeys (Macaca radiata radiata) of both sexes weighing between 2.5 and 4 kg were used in these experiments. Their platelet, reticulocyte and total leukocyte counts and the blood concentrations of hemoglobin and plasma proteins and the serum concentrations of aspartate amino transferase, alanine amino transferase and calcium are similar to the values reported for M. mulatta. Plasma glucose is lower when compared with reported values of M. mulatta and M. fascicularis. Insulin levels are comparable with those of M. mulatta and M. nigra. Histology of islets is similar to that of humans. Ovoid cell collections of islet cells are scattered throughout the pancreas. Ultrastructure of A, B and D cells is similar to humans. These findings suggest that this relatively underutilized macaques may be a suitable model for biomedical research.

Intravenous glucose tolerance test in Macaca radiata radiata.

A reliable method for performing sensitive intravenous glucose tolerance tests in monkeys has been standardized. This helps in assessment of beta cell function. A normal curve for glucose disposal is constructed. A high variability in insulin levels is also documented.

Effects of sodium lactate infusion on cisternal lactate and carbon dioxide levels in nonhuman primates.

OBJECTIVE: To further the understanding of lactate-induced panic in patients with panic disorder, the authors examined cisternal lactate and carbon dioxide levels in nonhuman primates after infusions of sodium lactate comparable to those used in studies of human beings. METHOD: CSF and venous blood lactate, pH, PCO2, PO2, and bicarbonate were measured in five ketamine-anesthetized nonhuman primates, without mechanical ventilation, before and after they underwent infusions of sodium lactate. In addition, the same measurements were made for three of the five subjects who were given saline infusions. RESULTS: Despite the development of the characteristic peripheral biochemical effects of infused sodium lactate--increased lactate and bicarbonate levels and metabolic alkalosis--no increases in central lactate or carbon dioxide levels were observed. Saline infusions produced no biochemical effects on venous and cisternal measures. CONCLUSIONS: The results of this study are in keeping with previous findings of nonpermeability of the blood-brain barrier to anionic compounds such as lactate. They therefore support theories of lactate panic based on cognitive and/or brainstem misevaluation of peripheral somatic sensations.

An avidin-biotin micro-enzymeimmunoassay for monkey chorionic gonadotrophin.

The avidin-biotin micro-enzymeimmunoassay used antiserum to the beta-subunit of sheep LH, biotinylated hCG as tracer and pooled serum from pregnant monkeys as a reference preparation of monkey CG. The assay was validated for specificity, by checking serum samples known to contain high quantities of LH, and false positive results were eliminated to a great extent by prior heating of serum samples at 60 degrees C for 30 min. In 45 bonnet monkeys the test gave 10 true positives, 34 true negatives and 1 false positive, i.e. an accuracy of 97%. The serum profiles of CG, measured by the EIA, in bonnet, cynomolgus and rhesus monkeys during early pregnancy were very similar to those reported for bonnet and rhesus monkeys when a liquid-phase radioimmunoassay was used.

Effect of LHRH agonists and antagonists in male and female bonnet monkeys (Macaca radiata).

A few analogues of LHRH have been tested in the adult bonnet monkeys using change in serum testosterone following LHRH injection as a parameter of response to LHRH. Of the four analogues tested in male monkeys, Buserelin was found to be the most potent one in increasing serum testosterone levels. Injection of the LHRH antagonist at 1600 h resulted in the abolition of the characteristic nocturnal surge of testosterone observed in adult bonnet monkeys maintained under regulated light conditions. Following administration of LHRH a/s during early pregnancy, serum chorionic gonadotropin levels decreased though the course of pregnancy was not affected. These results suggest that bonnet monkey can be successfully employed to test LHRH analogues.

Serum concentrations of chorionic gonadotrophin, oestradiol-17 beta and progesterone during early pregnancy in the south Indian bonnet monkey (Macaca radiata).

Serum concentrations of chorionic gonadotrophin (CG) during early pregnancy in the bonnet monkey (Macaca radiata), were determined by a radioimmunoassay validated for measuring monkey chorionic gonadotrophin. The earliest time at which CG could be detected was on the 28th day of a fertile cycle and it could not be detected beyond the 50th day. During a fertile cycle, mean values of oestradiol-17 beta, in addition to exhibiting a preovulatory peak around Days 9-10 of cycle, exhibited a secondary peak around Day 30 of the cycle. Serum progesterone levels remained elevated throughout the period of study from Day 18 to Day 50 and at no time did the levels fall below 2 ng/ml.

Studies on the blood group antigen Ina.

Anti-Ina antibodies have been observed in 30 of the 41 anti-Rh donors hyperimmunised with group O Ror In(a+) blood. They have also been found in four of 60 Rh immunised women. In three of these the husbands and previous children were In(a+). However, there was no evidence of haemolytic disease of the newborn due to anti-Ina antibodies alone. The Ina antigen is developed at birth though it is weaker than in adults. The strength of the Ina antigen decreases during pregnancy and returns to the previous level three months afer termination of pregnancy. The Ina antigen is affected by papain, trypsin, bromelin and neuraminidase. The antibodies in all the donors, even though their sera were preserved for more than 10 years at -20 degrees C, reacted in saline and albumin milieux and in the indirect antiglobulin procedure using anti-IgG reagents. These antibodies are complement binding as shown by the two-stage complement binding test. Treatment with 2-mercaptoethanol and DEAE-column chromatography suggested the anti-Ina antibodies were of the IgG class. Anti-Ina antibodies could possibly cause a transfusion reaction since In(a+) red cells when transfused into individuals possessing anti-Ina antibodies are eliminated from the circulation within 20 minutes.

Blood groups of macaques:a comparative study.

Distribution of the human-type and of the simian-type blood groups in rhesus, crab-eating, bonnet, pig-tailed and stump-tailed macaques revealed significant similarities and differences among these species. Human-type A--B-O blood groups cut across taxonomic lines and seem less value for taxonomic purposes than the simian-type blood groups detected by cross-reacting isoimmune rhesus monkey sera.

Blood groups of bonnet macaques (Macaca radiata), with a brief introduction to seroprimatology.

The human-type A-B-O blood groups of 52 bonnet macaques (Macaca radiata) were determined. Application of method of population genetics indicated the gene frequences to be 0 = 0.173, a = 0.480 and B = 0.347. Cross testing of sera and red cells of the bonnet macaques revealed two blood-type-specific isoagglutinins, one of them strong enough for use as a blood typing reagent. No blood group polymorphism was revealed by testing bonnet macaque red cells with isoantisera produced in rhesus monkeys (M. mulatta) and in crab-eating macaques (M. fascicularis). The rhesus and crab-eating macaque isoantisera reacted either with all or with none of the bonnet macaque red cells tested.

Effect of LH/FSH-RH on circulating serum gonadotropins in the bonnet monkey.

PIP: The effect of luteinizing hormone/follicle stimulating hormone-releasing hormone (LH/FSH-RH) on circulating serum gonadotropins was studied in the bonnet monkey. The RH, when administered either iv or im failed to elicit any marked increase in serum LH in 2 out of 3 monkeys. There was an increase in serum FSH levels in intact animals. In ovariectomized monkeys, a considerable increase in FSH levels was seen when the monkeys were pretreated with estrogen-progesterone combination but the treatment had no effect on the LH release by the RH.^ieng