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Laboratory rats are frequently used as animal models in research. Since the 1920s rats are bred and reared in laboratories which affects anatomy, physiology, and behavior responses. In the present study we exposed laboratory and wild rats to predator odor and comparatively analyzed their behavioral and physiological responses. In detail, Warsaw Wild Captive Pisula Stryjek (WWCPS) rats and Lister Hooded (LH) rats were exposed to the predator odor 2,3,5-trimethyl-3-thiazoline (TMT), their behavior was videotaped and blood samples were collected for subsequent serum corticosterone analysis. In both rat stocks, exposure to TMT induced avoidance behavior and increased freezing behavior. Notably, the increase in freezing was based on an increase number of freezing events in LH rats whereas WWCPS rats prolonged the mean duration of the single freezing events. Interestingly, TMT exposure lead to a serum corticosterone increase in WWCPS rats but not in LH rats. Furthermore, WWCPS rats generally expressed decreased but faster locomotor activity, as well as more grooming behavior than LH rats. Taken together, these data indicate differences in behavioral and physiological defensive responses to predator odors in the two rat stocks., (Copyright © 2018 Elsevier Inc. All rights reserved.)

The aim of this study was to study the effect of multipotent mesenchymal stromal cells (MMSC) transplantation on blood biochemical parameters after liver resection in Mature and old laboratory animals. The conducted studies show the ability of old laboratory animals to respond to MMSC transplantation after liver resection by reducing the severity of hepatocyte cytolysis. A comparative analysis of the biochemical parameters of Mature and old laboratory animals after liver resection should be noted a more pronounced response of mature animals in relation to the ability of the liver to synthesize urea.

This study determined the phases of sexual development of the male Mongolian gerbil (Meriones unguiculatus) based on an integrative analysis of testicular morphology, hormonal data and sperm parameters. Male gerbils were analysed at 1, 7, 14, 21, 28, 35, 42, 50, 60, 70, 90, 100 and 120 days of age. Body, testicular and epididymal weights increased up to Day 70, 60 and 90, respectively. The impuberal phase, characterised by the presence of gonocytes, extended until Day 14. The prepubertal period lasted until Day 42, when puberty was achieved and a drastic increase in serum testosterone levels, mature adult Leydig cells and elongated spermatids was observed. Gerbils at 60 days of age showed a remarkable number of spermatozoa in the testis, epididymidis caput/corpus and cauda, and at Day 70 the maximum daily sperm production was reached. However, the gerbil may be considered sexually mature only from Day 90 onward, when sperm reserves become stable. The total transit time of spermatozoa along the epididymis of sexually mature gerbils was 11 days, with 1 day in the caput/corpus and 10 days in the cauda. These data cover a lacuna regarding the reproductive parameters of this rodent and provide foundations for its use in testicular toxicology studies.

Background: Some factors such as sex, age, and captivity conditions have a direct influence on the normal hematological and serum biochemical parameters of African green monkeys. On the other hand, reliability in reported values is in many cases limited by studied animal number (<200) and there is not report on the correlation of these parameters with the age in each sex animal group. Thus, this study sought determining normal hematological (11) and serum biochemical parameters (9) of 400 captive housed African green monkeys and also correlate them with the age of the animals., Methods: A total of 200 females and 200 males were grouped by the sex and age groups (1-2, 3-4, 5-6, and 7-8 years old) for measuring normal values of hematological and serum biochemical parameters and to study the correlation of these parameters with the age of the animals., Results: As key outcome, the main hematological and serum biochemical reference values of African green monkeys were determined. Significant differences (P < 0.05) were found among 95% of studied parameters between males and females. About 75% and 95% of the parameters were influenced by the age in the female and male groups, respectively. About 35% of hematological and serum biochemical parameters correlated positively (R(2) > 0.5) with the age in the female monkeys. On the contrary in the male monkeys, only 45% of parameters correlated positively with the age (R(2) > 0.5)., Conclusions: Thus, authors believe that results of this study are important for assisting researchers in the assessment of health status of captive housed African green monkeys for preclinical studies., (© 2015 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.)

Xenopus laevis, the African clawed frog, is commonly used in developmental and toxicology research studies. Little information is available on aged X. laevis; however, with the complete mapping of the genome and the availability of transgenic animal models, the number of aged animals in research colonies is increasing. The goals of this study were to obtain biochemical and hematologic parameters to establish reference intervals for aged X. laevis and to compare results with those from young adult X. laevis. Blood samples were collected from laboratory reared, female frogs (n = 52) between the ages of 10 and 14 y. Reference intervals were generated for 30 biochemistry analytes and full hematologic analysis; these data were compared with prior results for young X. laevis from the same vendor. Parameters that were significantly higher in aged compared with young frogs included calcium, calcium:phosphorus ratio, total protein, albumin, HDL, amylase, potassium, CO2, and uric acid. Parameters found to be significantly lower in aged frogs included glucose, AST, ALT, cholesterol, BUN, BUN:creatinine ratio, phosphorus, triglycerides, LDL, lipase, sodium, chloride, sodium:potassium ratio, and anion gap. Hematology data did not differ between young and old frogs. These findings indicate that chemistry reference intervals for young X. laevis may be inappropriate for use with aged frogs.

Current methods for measuring acetylcholinesterase (AChE) activities in whole blood use butyrylcholinesterase (BChE)-selective inhibitors. However, the poor selectivity of these inhibitors results in the inhibition of AChE activity to some degree, leading to errors in reported values. The goal of this study was to develop and validate a simple assay for measuring AChE and BChE activities in whole blood from humans as well as experimental animals. Blood was fractionated into plasma and erythrocytes, and cholinesterase activities were titrated against ethopropazine and (-)-huperzine A to determine the lowest concentration of ethopropazine that inhibited BChE completely without affecting AChE activity and the lowest concentration of (-)-huperzine A that inhibited AChE completely without interfering with BChE activity. Results indicate that 20 µm ethopropazine can be successfully used for the accurate measurement of AChE activity in blood from humans as well as animals. Use of (-)-huperzine A is not required for measuring BChE activity in normal or 'exposed' blood samples. The method was validated for blood from several animal species, including mice, rats, guinea pigs, dogs, minipigs, and African green, cynomolgus and rhesus monkeys. This method is superior to all reported methods, does not require the separation of erythrocyte and plasma fractions, and is suitable for measuring cholinesterase activities in fresh or frozen blood from animals that were exposed to nerve agents or those that were administered high doses of BChE. The method is simple, direct, reproducible, and reliable and can easily be adapted for high-throughput screening of blood samples. Published 2012. This article is a US Government work and is in the public domain in the USA., (Published 2012. This article is a US Government work and is in the public domain in the USA.)

We sought to determine whether sex had a significant effect on the hematologic and serum chemistry analytes in adult sand rats (Psammomys obesus) maintained under normal laboratory conditions. According to the few data available for this species, we hypothesized that levels of hematologic and serum chemistry analytes would not differ significantly between clinically normal male and female sand rats. Data analysis revealed several significant differences in hematologic parameters between male and female sand rats but none for serum biochemistry analytes. The following hematologic parameters were greater in male than in female sand rats: RBC count, hemoglobin, hematocrit, red cell hemoglobin content, and percentage monocytes. Red cell distribution width, hemoglobin distribution width, mean platelet volume, and percentage lymphocytes were greater in female than in male sand rats. The sex of adult sand rats is a source of variation that must be considered in terms of clinical and research data. The data presented here likely will prove useful in the veterinary medical management of sand rat colonies and provide baseline hematologic and serum chemistry analyte information for researchers wishing to use this species.

Some environmental interventions can result in physiologic and behavioral changes in laboratory animals. In this context, the handling of adolescent or adult rodents has been reported to influence exploratory behavior and emotionality. Here we examined the effects of handling on memory and anxiety levels of adolescent rats. Male Sprague-Dawley rats (age, 60 d) were divided into a control group and a handled group, which were handled for 5 min daily, 5 d per week, for 6 wk. During handling bouts, the rat was removed from its cage, placed in the experimenter's lap or on the top of a table, and had its neck and back gently stroked by the experimenter's fingers. During week 6, each rat's anxiety level was evaluated in the elevated plus-maze (EPM) test. Learning and memory were evaluated 48 h later, by measuring escape latency in the elevated plus-maze test. Plasma corticosterone and catecholamine levels were measured also. Norepinephrine levels were lower in the handled rats compared with control animals, with no differences in epinephrine and corticosterone. As compared with the control rats, the handled rats showed increases in the percentage of time spent in the open arms of the test apparatus, percentage of entries into open arms, and number of visits to the end of the open arms and decreases in the latency of the first open arm entry. Escape latency was lower in the handled rats compared with control rats in both the first and second trials. The data obtained suggest that handling decreases anxiety levels and improves learning skills and memory in rats.

We validated the Coat-a-Count radioimmunoassay (RIA) kit for measuring testosterone in plasma samples of the subterranean rodent Ctenomys talarum and evaluated testosterone levels in free-living and captive individuals. The performance of the assay was evaluated by the assessment of parallelism, accuracy and precision. Moreover, the high specificity of the assay antibody was confirmed by high-pressure liquid chromatography with ultraviolet detector, followed by electrospray ionization tandem mass spectrometry. Results indicated that plasma samples have to be treated with heat and diluted before the RIA for the optimization of the assay. Plasma testosterone concentrations in free-living animals were outstandingly elevated (up to 329 ng/mL), which are among the highest ever reported for mammals. On the other hand, captivity produced a 14-fold decrease in plasma testosterone concentrations, emphasizing that very significant changes in the endocrine milieu may occur in wild animals kept under laboratory conditions. Our results place tuco-tucos as an interesting model for the study of androgen regulation in mammals, suggesting that target tissues may have low sensitivity to the testosterone signal and agree with a scenario of elevated levels of sex hormone-binding globulin in plasma., (© 2011 Wiley Periodicals, Inc.)

The South African clawed frogs Xenopus laevis and X. tropicalis are fully aquatic amphibians and well-established animal models. Because genetically engineered laboratory Xenopus are now being produced, the establishment of normal reference ranges for serum biochemical and hematologic parameters is essential for phenotyping and as a diagnostic aide. We determined normal reference ranges for hematologic values from 3 populations of X. laevis: wild-caught frogs (n = 43) and frogs from 2 commercial sources (A, n = 166; B, n = 109). For serum biochemistry, we determined normal reference ranges for frogs from source A and wild-caught frogs divided by sex and season. Significant differences across populations were found in WBC and RBC counts, hemoglobin concentration, hematocrit, mean corpuscular hemoglobin concentration, and mean corpuscular volume. Among serum biochemical analytes, significant differences were found for albumin:globulin ratio, anion gap, and concentrations of albumin, globulin, total protein, lipase, alanine transaminase, γ-glutamyl transpeptidase; creatine phosphokinase; indirect, direct, and total bilirubin; cholesterol, low-density lipoprotein lipase, carbon dioxide, glucose, lactacte dehydrogenase, calcium, chloride, and sodium. We hypothesize that these differences can be attributed to differences in water quality, habitat, ambient temperature, diet, sex, recent transport or shipment, and genetic background. However, testing that hypothesis is beyond the scope of the current study. In addition, clinical chemistry and hematologic reference range values Xenopus laevis are quite distinct from those for other species and are most consistent with the only values published for another fully aquatic amphibian, the Eastern hellbender (Cryptobranchus alleganiensis).

Babesia spp. are tick-transmitted apicomplexan hemoparasites that infect mammalian red blood cells. Our purpose was to determine the prevalence of Babesia infection in a colony of captive baboons and to evaluate potential experimental routes of the transmission of the hemoparasite. DNA was extracted from the blood of baboons and tested for infection with Babesia by PCR and primers that amplify the 18s rRNA gene of the parasite. The overall prevalence of infection of Babesia in the baboon population was 8.8% (73 of 830). Phylogenetic analysis of the sequenced DNA from 2 baboons revealed that the Babesia isolate found in captive baboons was a novel species most closely related (97% to 99%) to B. leo. Blood from a Babesia-infected donor baboon was inoculated intravenously, intramuscularly, or subcutaneously into 3 naive baboons. The intravenously inoculated baboon was PCR-positive at 7 d after inoculation; the 2 baboons inoculated by other routes became PCR-positive at 10 d after inoculation. All 3 baboons remained PCR-positive for Babesia through day 31. Baboons experimentally inoculated with the new Babesia isolate did not exhibit clinical signs of babesiosis during the experiments. We demonstrated that captive baboons are infected with a novel Babesia isolate. In addition we showed that Babesia can be transmitted in the absence of the organism's definitive host (ticks) by transfer of infected blood through intravenous, intramuscular, and subcutaneous routes to naive baboons., (Copyright 2011 by the American Association for Laboratory Animal Science)

Cardiac troponins have proved to be reliable blood biomarkers for identifying a variety of myocardial alterations in humans and animals. Recently, an ultrasensitive cTnI assay (Erenna IA) has been used to demonstrate increases in baseline cTnI resulting from drug-induced myocardial injury in rats, dogs, and monkeys, as well as to document baseline cTnI ranges in Sprague-Dawley (SD) rats. The present study was initiated to use the Erenna cTnI assay to further document baseline cTnI concentrations in normal control animals from multiple strains, including SD, Spontaneous Hypertensive (SHR), Wistar, Wistar-Kyoto (WKY), and Fisher strains. Baseline cTnI concentrations were quantified in all rats tested, and males had higher mean cTnI concentrations than females of the same strain. SHR males had the highest mean cTnI concentrations and the largest cTnI variability. Interestingly, cTnI concentrations increased in castrated SHR compared with unaltered male SHR, whereas cTnI concentrations decreased in ovariectomized SHR compared with unaltered female SHR. These results show significant differences in cTnI concentrations between strains, sexes, and noncardiac surgical alterations in control animals, and identify these as potential contributing factors to cTnI baseline variability that should be taken into account when using ultrasensitive cTnI as a biomarker to assess preclinical cardiotoxicity.

Biologic samples from 18 (12 female, 6 male) Siberian hamsters (Phodopus sungorus) representing an aged colony (17 to 27 mo) were examined. Values for CBC and serum biochemical parameters were determined, and macroscopic and microscopic pathologic evaluations were performed. Blood urea nitrogen levels were significantly higher in male (54.2 ± 14 mg/dL) compared with female (35.3 ± 22 mg/dL) hamsters and correlated histologically with a higher incidence of chronic glomerulonephropathy in males (5 of 6 males; 0 of 12 females). All 18 hamsters had histologic evidence of follicular mite infestation. Half (6 of 12) of the female hamsters showed cystic rete ovarii. Other histologic findings included thymic or thyroid branchial cysts (3 of 18), focal enteritis (2 of 18), and single cases of hepatic hemangiosarcoma, renal adenoma, subcutaneous mast cell tumor, cutaneous sebaceous adenoma, cutaneous trichofolliculoma, squamous papilloma of the nonglandular stomach, epididymal cholesteatoma, pyometra, and pituitary craniopharyngeal cyst. This study is the first published report of hematologic and serum chemical values for any population of Siberian hamsters and the first published report showing a potential male predisposition for chronic progressive glomerulonephropathy and a potential female predisposition for cystic rete ovarii.

To provide fundamental basis for the tree shrew models of human diseases, we examined and compared the physiological and biochemical indexes between wild and laboratory tree shrews. Blood samples were taken from 54 wild tree shrews that were housed in laboratory for 1-2 months, and from 54 first-generation of the laboratory tree shrews; each group had nearly equal male and female composition. Some of the first reported physiological and biochemical indexes were showed no significant differences between genders, and these indexes in laboratory tree shrews were as follows [medium (inter-quartile range) ]: CK 1449 (956) U/L, CTNI 5.94 (7.23) ug/L, TBA15.6 (19.7) μmol/L, FRUC 393.5 (80.8) μmol/L and LDL-C0.36 (0.32); and in the wild tree shrews, 986 (564) U/L, 4.01 (4.10) μg/L, 20.0 (20.6) μmol/L, 379.0 (104.0) μmol/L and 0.46 (0.23) mmol/L, respectively. In the laboratory tree shrews, the variations of physiological and biochemical indexes were smaller, but the mean values of some indicators related to liver and heart functions became higher. These data would be valuable for the development of tree shrew models of human diseases.

Background: Animals in captivity should receive adequate sunlight exposure for sufficient generation of vitamin D [25(OH)D]. In the present study, 25(OH)D serum levels of 84 Callithrix penicillata primates were evaluated., Objectives: To determine 25(OH)D levels of those animals; to evaluate the influence of gender and period of sunlight exposure on their 25(OH)D levels., Methods: Three groups were evaluated: group 1 (n = 29) on free sunlight exposure; group 2 (n = 34) on partial sunlight exposure; group 3 (n = 21) without sunlight exposure., Results: The obtained 25(OH)D values were: group 1, 121.2 +/- 33.3 ng/ml; group 2, 115.2 +/- 32.2 ng/ml; group 3, 53.3 +/- 10.4 ng/ml. Significant statistical differences were obtained between groups 1 and 3 (p < 0.001) and groups 2 and 3 (p < 0.001); no statistical difference was found between genders., Conclusion: Direct sunlight exposure is essential for 25(OH)D sufficiency and it is proposed that the 25(OH)D normal range for captive Callithrix penicillata would be from 104.8 to 137.1 ng/ml (CI = 95%).

Ecophysiological research aiming at explaining the causes and consequences of variation in individual condition, health state, and allostasis is traditionally performed on captive animals under controlled laboratory conditions. The question about how captivity per se affects studied parameters is therefore of central importance for generalizing the information gained from such studies. We addressed this question by comparing various indexes of physiological condition of wintering greenfinches sampled in the wild and kept in captivity for different time periods. Bringing wild greenfinches into captivity did not result in systematic alteration in nine of 12 physiological parameters studied. Captive birds had consistently lower plasma carotenoid and uric acid levels than wild ones. Variation in differential leukocyte counts did not reveal any signs of elevated stress of birds kept in captivity. These results indicate that for a number of physiological parameters, information obtained from captive animals can be generalized to natural situations. Variance in traits most closely related to physical exercise capacity (body mass and hematocrit) were much lower in the wild than in captivity. These findings suggest that under harsh environmental conditions experienced by wild birds (i.e., predation threat, scarce resources), traits such as hematocrit and body mass are fine tuned by physiological trade-offs.

Although growing number of evidences supports the gender differences of hemorheological variables in human, little is known about this question in experimental animals. Investigating this question and giving laboratory-specific reference values, blood samples from healthy male and female CD rats and beagle dogs were tested for hematological parameters (microcell counter), blood and plasma viscosity (capillary viscometer), erythrocyte deformability (bulk filtrometry and ektacytometry), erythrocyte aggregation (light transmission aggregometer) and fibrinogen concentration (coagulometer). Besides the inter-species alterations we found hematological gender differences too regarding the red blood cell count and hematocrit. In rats the erythrocyte aggregation index was higher in females, while in beagle dogs the males showed significantly higher aggregation index values. In rats the red blood cell deformability showed better values in females. In beagle dogs this was the opposite, the males had better elongation index (EI) values in the function of shear stress (SS). Besides this it was also experienced that the shape of the canine EI - SS curves differed from the rats' values. Gender differences with inter-species alterations can be observed in laboratory animals (rats and dogs) that have to be taken under consideration during planning experiments and evaluating results.

The postprandial hypertriglyceridaemia (PHT) rabbit, developed as a new animal model of metabolic syndrome, is characterized by PHT, central obesity and glucose intolerance. For detailed investigation of lipid metabolism characteristics in PHT rabbit, the plasma levels of apolipoproteins A-I, B, C-II, C-III and E were measured. Movements of apolipoproteins B100 and B48 were investigated using sodium dodecyl sulphate-polyacrylamide gel electrophoresis to determine whether postprandially increased triglyceride is exogenous or endogenous. The level of apolipoproteins A-I, B, C-II and E were increased in PHT rabbit after feeding. Apolipoproteins B100 and B48 were detected in the plasma fraction of d < 1.006 g/mL of the PHT rabbit. The postprandial increase in apolipoprotein B in the PHT rabbit reflects a numerical increase in lipoprotein particles in the blood; the increase in apolipoproteins C-II and E suggests some disturbance in lipoprotein catabolism. Apolipoprotein B48 was detected postprandially in PHT rabbits. These results suggest that delayed catabolism of exogenous lipids caused the retention of chylomicron remnants in the blood. Results also suggest that activities of the lipolytic enzyme lipoprotein lipase and hepatic triglyceride lipase were deficient and that the hepatic uptake of exogenous lipoproteins was delayed in the PHT rabbit. Especially, for examining remnant hyperlipoproteinaemia in humans, PHT rabbit is an excellent animal model for hypertriglyceridaemia research.

As our survey found that some commercial standard diets for laboratory animals contained oil with chemical properties of highly deteriorated oil, their influence should be checked on animals especially of very young age. Three-week old weanling Wistar rats were fed a commercial standard diet (commercial standard diet group) or AIN93G (AIN93G group), defined diet, for 7 weeks. Then both groups were fed AIN93G for 12 weeks. As a result, all the rats grew normally without diarrhea, seborrhea, dermatitis, or excessive hair loss through the feeding period and there was no significant difference in body weight increase, feed ingestion amounts and organ weights. But fecal excretion was high at 10 weeks of age when the diet was switched from the commercial standard diet to AIN93G. Although serum aspartate aminotransferase (AST) and alanin aminotransferase (ALT) of the commercial standard diet group were not statistically higher than those of the AIN93G group, two rats out of 8 in the former group showed dark-red patches on the liver surface, and necrosis in histological analysis. In addition, slight fatty degeneration of all the liver, and swelling tubuler epithelium of kidneys were also found in the group. Serum levels of triacylglycerol (TG), glucose (GLC) and total cholesterol (T-CHO) were high, and free fatty acids (NEFA), low in the commercial standard diet group in good accordance with our previous result from the study on ingestion of deteriorated oil. In conclusion, product specifications of oil in commercial standard diets should be laid down to pursue a reliable animal experimentation.

The simulation of human serum levels is essential in animal models to extrapolate the experimental results to clinical practice. Administration of a nephrotoxic drug such as cisplatin can be used to cause renal dysfunction as an approach to mimic human serum levels of renally excreted drugs. We aimed to determine the dose of cisplatin that did not affect the survival rate of mice and to achieve human-like serum concentrations of cefepime. Different doses of cisplatin (0, 10, 14, 18, 22 and 26 mg/kg) were given by intraperitoneal (i.p.) injection to mice three days prior to the i.p. administration of 80 mg/kg cefepime. With cisplatin doses of 18 and 22 mg/kg, the half-life of cefepime was significantly prolonged (P < 0.001) and all mice survived. The pretreatment with 26 mg/kg cisplatin significantly decreased survival (P = 0.001), but the half-life of cefepime was not significantly longer than of 18 mg/kg cisplatin. Serum levels of cefepime after the pretreatment with 18 mg/kg cisplatin were comparable to published human data. The administration of cisplatin appears to be a suitable method in mice for simulating human serum concentrations of renally excreted drugs.

This study directly demonstrates that cardiac troponin I (cTnI) is a sensitive, specific, and persistent biomarker in laboratory animals. Histopathological and pathophysiological cardiac changes in dogs, rats and mice correlated with increased serum cTnI with various cardiac inotropic agents, and cardiotoxic drugs and with cardiac arrhythmias, tachycardia, cardiac effusion with dyspnoea, and ageing. A comparison of six immunoassays for cTnI and cardiac troponin T (cTnT) to detect and monitor cardiac injury in a rodent model indicated that enzyme-linked immunosorbent (Life Diagnostics Inc and TriChem Resources Inc, West Chester, Philadelphia, USA) and Immulite (Diagnostic Products Corporation, Llanberis, UK) assays had low sensitivity and less than 1% of the dynamic range of Centaur (Bayer Healthcare Diagnostics, Newbury, UK) cTnI and Elecsys (Roche Diagnostics, Basel, Switzerland) and M8 (Bioveris Europe, Whitney, UK) cTnT assays. In dogs, however, the Immulite assay was effective and correlated with the Centaur. Serum concentrations were highly correlated but 10-fold lower for cTnT compared with cTnI with cardiac injury. Centaur assay also detected cTnI in myocardium from marmosets, swine, cattle, and guinea pigs, indicating it to be candidate cardiac biomarker for these species as well. Purified rat cTnI was 50% more reactive than purified human cTnI in the Centaur assay. In the rat, an age- and gender-dependent variation in serum cTnI was found. Male rats aged six and eight months had a 10-fold greater serum cTnI than age-matched females and three-month-old rats. These increases correlated with minimal histopathological change. Isoproterenol-induced serum cTnI increased up to 760-fold the minimal detectable concentration of 0.07 microg/L, within 4-6 h and decreased with a half-life of 6 h, with an expected return to baseline of 60 h. Severity of histopathological change correlated with serum cTnI during the ongoing injury.

Thyroid and embedded parathyroid glands were surgically removed (thyroparathyroidectomized) from adult chinchillas (Chinchilla laniger) to create an animal model of hypothyroidism. Thyroxine (T4) levels were measured at the time of surgery and one or two times after surgery from 10 thyroparathyroidectomized chinchillas and five sham controls to establish baseline serum T4 levels and to assess the degree and duration of hypothyroidism in this animal model. Baseline T4 levels ranged from 3.4 to 6.4 microg/dl (mean +/- 1 standard deviation, 5.25 +/- 0.84 microg/dl), with no differences between male and female chinchillas (5.4 +/- 0.6 microg/dl versus 5.2 +/- 1.0 microg/dl, respectively). T4 levels were significantly reduced in 80% of thyroparathyroidectomized chinchillas when measured 6 to 14 days after surgery, but reductions were variable, ranging from 9 to 89% in individual animals. There was rapid regrowth of thyroid tissue and a return of T4 levels to the baseline range in five of the seven animals followed for 1 to 2 months after surgery. T4 levels increased significantly in the sham-operated chinchillas, indicating a nonspecific effect of surgery. The results establish surgical procedures for creating a model of variable, transient hypothyroidism in the chinchilla. We also summarize published basal T4 values for various laboratory animals, to provide a convenient reference.

We showed previously that hepatic vitamin A concentrations of captive rhesus monkeys (Macaca mulatta) are subtoxic to toxic, with livers exhibiting stellate cell hypertrophy and hyperplasia. Although marmoset (Callithrix jacchus) livers are also high in vitamin A, no stellate cell irregularities were observed. To further characterize the effects of high dietary vitamin A from preformed sources, stored serum samples were analyzed from monkeys used for biomedical research and housed at the Wisconsin Primate Research Center. The monkeys had been fed commercially available monkey diets, providing vitamin A (as retinyl acetate) at levels exceeding NRC recommendations by a factor of four. The serum from both rhesus and marmoset monkeys had total serum vitamin A (retinol, retinyl esters and metabolites) within the expected range for both species, i.e., 1.44 +/- 0.34 and 1.41 +/- 0.72 micromol/L serum for rhesus and marmoset monkeys, respectively. However, high serum retinyl ester concentrations as a percentage of total serum vitamin A were present in both species, 12 +/- 5.1% (range, 5.5-23%) for rhesus and 27 +/- 14% (range, 10-57%) for the marmosets. Serum retinol concentrations were normal, i.e., 1.21 +/- 0.28 (rhesus) and 0.92 +/- 0.43 micromol/L (marmoset), compared with published values.

Typically, pharmacokinetic studies in mice require one animal per time point, thus resulting in differences due to dosing error, animal to animal variation and more importantly the euthanasia of a large number of animals. A method for the determination of pharmacokinetic data from serially bled mice to support early drug discovery is described. Sample analysis relies on liquid chromatography coupled with tandem mass spectrometry permitting robust and reproducible analysis requiring approximately 3 min per sample. Several parameters are discussed including the method of sample collection, preparation and analysis. The use of serially bled mice has lead to a remarkable reduction in animal usage and a corresponding reduction in compound required for such experiments. Using conventional methodology, a nine-point pharmacokinetic curve with four animals per time point would require 36 mice. With the method described below, only four mice in total are used and euthanasia is not required, permitting reuse after several weeks recovery and washout. Also, pharmacodynamic-pharmacokinetic correlation is possible and is demonstrated using a mouse model of diabetes.

This article is the result of an initiative between the European Federation of Pharmaceutical Industries Associations (EFPIA) and the European Centre for the Validation of Alternative Methods (ECVAM). Its objectives are to provide the researcher in the safety evaluation laboratory with an up-to-date, easy-to-use set of data sheets to aid in the study design process whilst at the same time affording maximum welfare considerations to the experimental animals. Although this article is targeted at researchers in the European Pharmaceutical Industry, it is considered that the principles underpinning the data sets and refinement proposals are equally applicable to all those who use these techniques on animals in their research, whether in research institutes, universities or other sectors of industry. The implications of this article may lead to discussion with regulators, such as those responsible for pharmacopoeial testing. There are numerous publications dealing with the administration of test substances and the removal of blood samples, and many laboratories also have their own "in-house" guidelines that have been developed by custom and practice over many years. Within European Union Directive 86/609EEC1 we have an obligation to refine experiments to cause the minimum amount of stress. We hope that this article will provide background data useful to those responsible for protocol design and review. This guide is based on peer-reviewed publications whenever possible, but where this is not possible we have used "in-house" data and the experience of those on the working party (as well as helpful comments submitted by the industry) for a final opinion. The guide also addresses the continuing need to refine the techniques associated with the administration of substances and the withdrawal of blood, and suggests ways of doing so. Data-sharing between laboratories should be encouraged to avoid duplication of animal work, as well as sharing practical skills concerning animal welfare and scientific problems caused by "overdosing" in some way or another. The recommendations in this guide refer to the "normal" animal, and special consideration is needed, for instance, during pregnancy and lactation. Interpretation of studies may be confounded when large volumes are administered or excessive sampling employed, particularly if anaesthetics are used., (Copyright 2001 John Wiley & Sons, Ltd.)

Common laboratory rodents have always been a favorite choice as a pet. Although diagnostic clinical pathology has not been viewed as practical for the rodent patient, current advances in technology make processing of small samples possible. Cultivation of the technical skills necessary for rodent sample collection has the potential to improve the standard of rodent veterinary care. This article provides an overview of rodent sample collection techniques, hematology, clinical biochemistry, serology, and clinical pathology of other tissues and fluids for laboratory rodents. General principles of clinical pathology can be applied across species. This article emphasizes the subtleties of the different rodent species which may impact diagnostic interpretation.

Aggressive territorial behavior was studied in 15 colonies of wild rats (Rattus norvegicus), each consisting of 2 males and 1 female. One of the males attacked an intruder rat more frequently and had a higher body weight than the less aggressive one. In another experiment, male and female rats were raised in colonies from weaning to adulthood. Animals were weighed every 7 days until 90 days of age when plasma testosterone was measured in males, and plasma glucose, hepatic and muscle glycogen were measured in both males and females. THe heavier (and thus possibly dominant) males in the colonies of 3 males and 1 female also had a higher body weight than males raised with females, but without any male partner. In this long-term social relationship there were no significant differences in carbohydrate metabolism among the animals. The differential growth rate among males was established around the period of sexual maturity. Moreover, when adult, heavier males had higher plasma testosterone levels compared to the other members of the colony and also to males that had no other competitive male partner. This higher androgenic hormone level may be one of the causal factors involved in the weight increase of the dominant male in the colony.

Blood samples from the right ventricle (RV) in rats are usually assumed to be representative of mixed venous blood. However, results presented here suggest that this assumption may not be true in all experimental circumstances. Nineteen male Wistar rats were anaesthetized with urethane and mechanically ventilated. The six gases used in the multiple inert gas elimination technique (MIGET) were administered continuously and samples of mixed expired air and blood from the left carotid artery and RV were simultaneously withdrawn. While inert gas concentrations in arterial blood and mixed expired air were almost homogeneous, those obtained from RV blood showed a high variability, specially evident for the less soluble gases in blood. As inert gases are cleared in the lungs according to their solubility in blood and they are only replaced through the lower systemic circulation, higher or lower concentrations found in RV samples than those expected suggests a preferential collection of blood from the lower or upper systemic circulation, respectively.

We determined the effects of sex, age, and environment (inbred, captive-wild, and wild animals) on selected blood parameters of the cotton rat (Sigmodon hispidus) in central Oklahoma (USA) from 1990 to 1994. Male and female cotton rats had similar blood profiles. Age-related differences were confined to differential white blood cell counts where adults possessed greater numbers of neutrophils and lower numbers of lymphocytes compared to juveniles. Environment had a strong influence on many hematological parameters. Hematocrit, hemoglobin concentration, RBC count, and eosinophil number were generally greater for wild stocks compared to inbred animals, and differences were more pronounced for adults than juveniles.

Close correlation of oxygen-binding properties of haemoglobin and the content of malonyl dialdehyde (MDA) in animal blood plasma was shown. The higher the affinity of haemoglobin to O2, the lower MDA level in the blood plasma. For the muskrat Ondatra zibethica, representing diving animals, MDA content is lower and catalase activity is higher compared with the laboratory rat.

Since thromboxane (TX) B2 is not a reliable indicator of TXA2 generation in vivo, because of artifactual TXA2 generation during blood collection, we tested the feasibility of replacing TXB2 with 11-dehydro (dh)-TXB2 as the indicator. Plasma levels of TXB2 and 11-dh-TXB2 were measured after i.v. administration of TXB2 to rabbits, guinea pigs, rats, dogs and a monkey, and after i.v. infusion of collagen in rabbits. In the rabbits and dogs, 2,3-dinor-TXB2 levels were also measured. After intravenous injection of TXB2 (10 micrograms/kg) in rabbits, the ratio of the area under the curve (AUC) of 11-dh-TXB2 to that of TXB2 (1.94) was far higher than the AUC ratio between 2,3-dinor-TXB2 and TXB2 (0.42). When endogenous TXA2 was generated by infusion of collagen (2 mg/kg/5 min), the plasma level of 11-dh-TXB2 was significantly increased, and had a longer half-life than TXB2. In the guinea pigs, rats and monkey, the peak plasma levels of 11-dh-TXB2 were significantly increased after injection of TXB2 (10 micrograms/kg, i.v.), whereas the AUC ratios of 11-dh-TXB2/TXB2 were less than one fourth of that in rabbits. No significant increase in 11-dh-TXB2 was observed after TXB2 injection (10 micrograms/kg) in dogs, but the 2,3-dinor-TXB2 level rose significantly, its AUC ratio to TXB2 being 0.29, comparable with that in rabbits. The order of the 11-dh-TXB2/TXB2 AUC ratios was: rabbits > guinea pigs > monkey > rats >> dogs.(ABSTRACT TRUNCATED AT 250 WORDS)

Repeated episodes of diarrhoea were seen in 4 laboratory beagles after experimental renal surgery and feeding a modified diet. Small intestinal bacterial overgrowth (SIBO) was suspected by exclusion of other causes and measurement of plasma folate. SIBO was confirmed by quantitative duodenal bacteriology. Beagles with SIBO can show no clinical signs, experimental stress and dietary change may have been reasons why these 4 beagles exhibited clinical signs with SIBO. Despite normal gut histology an increase in gut permeability was found using sugar absorption tests. This increased permeability had the potential to cause variations in drug absorption during experimental studies.

A systematic programme of comparative nuclear magnetic resonance measurements of the membrane permeability for water (Pd) and of activation energy (Ea,d) of this process in red blood cells of various wild, laboratory and domestic animals was carried out here. The RBC from humans, cow, sheep and kangaroos had Pd values around 5 x 10(-3) cm/s at 25 degrees C, 7 x 10(-3) cm/s at 37 degrees C with Ea,d values around 25 kJ/mol. For RBC from other ten marsupial species and from mouse, rat and rabbit, the Pd values were more than twice as high as for human RBC. For mosr RBC a high value of Pd was associated with a low value of Ea,d (range from 15 to 21 kJ/mol), pointing to specialized channels for water diffusion incorporated in membrane proteins. Recently a channel-forming integral protein of 28 kDa (CHIP 28) was identified as a major water channel protein in the RBC membrane. A procedure for quantitating the purified CHIP 28 by densitometry of silver-stained polyacrylamide gel electrophoreograms was developed. The analysis of a purified fraction of CHIP 28 showed that the 28 kDa component represents approximately two-thirds of the sample with the remainder comprising the glycosylated high-molecular-weight component. A correlation between the content in CHIP 28 and the relative water permeability among RBC from different vertebrate species was attempted.

During the conduct of a long-term toxicity/reproduction study using rhesus monkeys (Macaca mulatta), periodic hematologic and serum biochemical analyses were undertaken on blood obtained from 15 untreated (vehicle control) adult female monkeys over a period of 3 yr, 20 untreated adult males over a period of 6 yr, and 9 infants (5 male, 4 female, whose dams were the vehicle control females) from 12 wk through 2 yr of age. All samples were obtained by femoral puncture. To facilitate handling, ketamine hydrochloride (Ket.HCl) was administered to the males, and for some of the later infant samplings. Complete blood cell counts were performed using electronic cell counters, while reticulocyte counts and leukocyte differentials were assessed manually. The serum biochemistry analyses were performed with automated analyzers. These data were obtained as part of a health monitoring program, with medians and 2.5 to 97.5 percentiles for each parameter indicated. Some monitored parameters were found to be affected by the type of equipment used and by the use of Ket.HCl.

We analyzed 161 raccoon (Procyon lotor) blood samples obtained from New Jersey (n = 109), rural Pennsylvania (n = 29) and laboratory confined animals (n = 23) in the USA for lead content; we found significantly higher levels in the New Jersey raccoons (mean = 4.4 micrograms/dl, SE = 2.9). There was no difference between the lead levels of raccoons from the other two groups (mean = 2.6, SE = 0.5 and mean = 2.5, SE = 0, respectively).

An ELISA was developed to determine the reactivity of peroxidase labelled Protein A and a recombinant Protein A + Protein G construct, to sera from a variety of laboratory, domestic and wild animals from Africa. There was variability in the binding capacity of sera from individuals of the same species, but four groups could be recognized. Sera from birds and crocodiles were at most weakly reactive with either Protein A or the chimeric construct. Sera from some domestic animals such as horse, goat and cat, and sera from some wild ungulates including buffalo, wildebeest, waterbuck and impala were reactive with Protein A, but reacted to a much greater degree with the chimeric construct. Sera from larger wild animals such as elephant, rhinoceros and giraffe were strongly reactive with the chimeric protein and moderately reactive with Protein A. Sera from primates and dog, pig, guinea pig and rabbit reacted strongly with both proteins. Chimeric proteins that combine the IgG binding capacities of Protein A and Protein G can be used to detect immunoglobulin from a wide variety of African wild animal species. They may thus be of great value in seroepidemiological investigations of these animal populations.

1. The concentration of lipoproteins and lipids in the sera of several species of healthy adult laboratory animals (guinea-pig, rabbit and rat), domestic animals (cattle, sheep, goat, horse and swine) and wild animals (deer, wild boar, mink and fox) have been investigated under physiological conditions. Special attention was paid to the lipoproteins which are precipitated with heparin-MnCl2 and to the ratio of lipids in separated alpha and beta lipoproteins. 2. Most of the studied animals had significantly lower concentrations of total lipoproteins, beta lipoproteins and cholesterol than those in man (P < 0.001). Only some wild animals (mink and fox) had concentrations of total lipoproteins, beta lipoproteins, phospholipids and cholesterol which were significantly higher than man's (P < 0.001). 3. The ratio of lipids in the separated lipoprotein fractions also differed between the examined animals. Thus, in the sheep, rabbit, guinea-pig and wild boar most of the cholesterol (70-76%) was in beta lipoproteins. However, in the horse (60%), mink (60%), fox (65%), goat (70%) and cattle (73%) the high density lipoprotein fraction was the main carrier of cholesterol. 4. With the exception of the guinea-pig (42%) most of the serum phospholipids were found in the high density lipoprotein fraction.

Plasma endotoxin levels were examined in laboratory mice and rats sustained under germ-free (GF), SPF and conventional (CV) conditions. The values were 1.72 (GF mouse), 2.67 (SPF mouse), 2.44 (CV mouse), 2.05 (GF rat), 2.56 (SPF rat), and 3.13 pg/ml (CV rat). The GF animals showed the lowest levels, followed by SPF and CV animals, in that order. When SPF animals were transferred into the conventional condition (conventionalization), the plasma endotoxin levels changed slightly, but not significantly. The present study demonstrates that the levels of plasma endotoxin in healthy laboratory animals are relatively stable and are not affected by microbiological conditions.

The effect of various confinement conditions on physical fitness in dogs was evaluated. Eighteen 9.5- to 10-month-old female purpose-bred Beagles were maintained individually for 3 months at a time in 1 of 6 confinement conditions: Condition A--an outdoor housing area with a conventional dog house and free access to a 6.1 x 9.1-m pen; condition B--outdoor kennel with a conventional dog house and free access to a 1.8 x 6.1-m run; condition C--indoor environmentally controlled 1.2 x 3.66-m run; condition D-0.9 x 1.2 x 0.84-m conventional laboratory cage in an indoor environmentally controlled room; condition E--0.9 x 1.2 x 0.84-m conventional laboratory cage in an indoor environmentally controlled room with treadmill exercise (7 km/h at a 10% grade) for 30 min/d, 5 d/wk; condition F--0.71 x 0.86 x 0.69-m conventional laboratory cage in an indoor environmentally controlled room. During the final week of each 3-month interval, muscle succinate dehydrogenase enzyme activities and submaximal exercise heart rates (during treadmill exercise) were determined to estimate physical fitness. Also, 5 days after being moved into a different housing condition, blood samples were collected for plasma cortisol determination. The type of confinement condition for dogs had little effect on muscle succinate dehydrogenase activity, but had a modest effect on submaximal exercise heart rates of dogs.(ABSTRACT TRUNCATED AT 250 WORDS)

The staining characteristics of the peripheral blood cells from mouse, rat, guinea pig, rabbit, dog, marmoset and monkey were studied. In marmoset, it is easy to distinguish neutrophils from eosinophils by using the phosphate-buffered solution of pH 5 or 6. It was found in the special staining methods that neutrophil granules showed intense peroxidase and Sudan black B reactions in marmoset in comparison with those in the other species of experimental animals. Neutrophil granules rabbit was, however, intensely stained with esterase and acid phosphatase.

Environmental enrichment devices are a potential way to enhance psychological well-being in laboratory animals. The effects of such devices need to be systematically evaluated before they are recommended for widespread use. The purpose of this research was to monitor the behavioural and physiological responses of adult female rhesus macaques to a simple enrichment device. The apparatus consisted of a box attached to the monkey's home cage that contained a radio and a food dispenser, which could be controlled by the monkeys via contact detectors. Radio and food dispenser use were automatically recorded. Whole blood serotonin (WBS), plasma cortisol and abnormal behaviour were measured in 5 monkeys before, during and after a 20-week period in which the monkey's cages were equipped with the device. All monkeys used the device (3 of the 5 subjects earned an average of more than 200 food pellets per day). Mean plasma cortisol and whole blood serotonin did not differ across sampling times, suggesting that the apparatus had no effect on basal stress levels. There was an inverse relationship between apparatus use and cortisol levels in 76% of the samples, but only 3 of 17 coefficients were significant. There was a significant but small negative correlation between apparatus use and self-abusive behaviour. This enrichment device was readily used by adult rhesus monkeys and could be adapted for use in a wide variety of laboratory settings.

Haemoglobin electrophoresis and chromosome analysis indicate that the widely used multimammate rat (chamois coloured) as experimental animal belongs to the species Mastomys coucha (Smith, 1836) and not, as commonly assumed, to Mastomys natalensis (Smith, 1834).

White adipose tissue biopsies and plasma samples were obtained from hibernating yellow-bellied marmots (Marmota flaviventris) maintained in the laboratory. In addition, biopsies and plasma samples were obtained from normothermic animals in the field and laboratory. Measurement of plasma free fatty acid (FA) levels indicated that winter laboratory animals exhibited increased lipolysis. Additionally, analysis of white adipose tissue triacylglycerol revealed that the FA composition of the storage fat in animals maintained on the standard laboratory diet is remarkably simple and uniform between different adipose depots in the same animal. Three FAs (palmitic, oleic, and linoleic acids) made up greater than 95% of the total. Triene (alpha-linolenate) was found in newly captured animals, but the percentage of this FA decreased rapidly when the animals were maintained on the standard laboratory diet. Throughout the hibernation season (October to April), white adipose tissue-saturated FA percentage decreased, monoene percentage remained constant, and diene percentage increased. Analysis of plasma FA composition suggested that these animals tended to metabolize saturated FAs from stored lipid during hibernation and that dienes were mobilized briefly after the last arousal from hibernation in spring. From these observations, we hypothesize that marmots preferentially metabolize saturated fats during the hibernation period and that essential FAs of the omega 6 series tend to be metabolized more slowly than other FAs. These characteristics suggest that marmots are a valuable animal model in which to study lipid metabolism.

The influence of long term rearing conditions on the hematological and serum biochemical values in laboratory-bred female cynomolgus monkeys (Macaca fascicularis) was studied under the following two conditions. Group A: New born infants were nursed by their mothers for at least 12 weeks after birth. After weaning, four monkeys of approximately equal age and body weight were kept in a relatively small cage (90 cm W x 60 cm D x 60 cm H) and reared to five years of age. These rearing conditions are commonly used in the breeding program of our primate center. Group B: One male and five females of wild origin were introduced into a relatively large cage (180 cm W x 180 cm D x 160 cm H). Female monkeys born in these cages were used in this study. All of them were reared by their own mothers and grew well in these cages. Male monkeys born in these cages were removed at about 3.5 years of age. Hematological and serum biochemical values between the two groups in each age class were compared and analyzed by the canonical discriminant analysis (the discriminant analysis with reduction of dimensionality). Age correlated highly with the value of the first canonical variate. The second canonical variate detected a difference in the blood hematology and biochemistry probably induced by the difference in rearing conditions.