Deprivation or restricted access to either food or fluids is a common research procedure in laboratory animals. The purpose of the present review is to present and summarize some of the important physiologic effects of such procedures and to assess their effect on the well-being of the animal. This assessment is presented within a context of the typical research objectives of such procedures. Specific suggestions are made that are intended to strike a balance between meeting these research objectives and ensuring the physiologic and behavioral welfare of the animals under study. Most of the information presented is specifically related to rats and mice but, with appropriate adjustments, the principles likely will generalize to other laboratory species. I present evidence that after 12 to 24 h without access, animals efficiently reduce further fluid or energy losses by a combination of behavioral and physiologic adjustments. These adjustments likely minimize the additional physiologic or psychologic stress of deprivation. Animals have endogenous nycthemeral rhythms that make them particularly adaptable to once-daily occurrences, such as food or water access. Longer periods of acute deprivation or chronic restriction are acceptable procedures, but only with suitable monitoring protocols, such as routine weighing and target weights. In the case of chronic food restriction, the use of species-, age-, and strain-specific target growth rates is more appropriate than using a fraction of age-matched free-fed animal weights as a target.

The relative amounts of the different forms of morphine, and many other pharmacologic agents, depend on temperature and pH. Some forms are more efficacious because they are uncharged and can penetrate lipid membranes more easily than the charged forms. Persons who administer pharmacologic agents to ectotherms (that is, cold-blooded animals) should consider the effect of temperature on the relative amounts of the different forms of drugs. For example, the fraction of morphine present in the uncharged form is twice as high in a fish or frog at 5 °C as in a mammal at 37 °C. Moreover, because the pH of blood, plasma, and tissues of ectotherms is higher when they are held at lower temperatures, the combined effect of temperature and pH on the speciation of pharmacologic agents also should be considered. In addition, the total solubility of morphine and other pharmacologic agents depends on temperature and pH. The purpose of this overview is to describe how temperature and pH influence the solubility and speciation of morphine.

We evaluated ketoprofen, a nonsteroidal anti-inflammatory drug (NSAID), as an antinociceptive supplement to chloral hydrate anesthesia in mouse. Effects of ketoprofen on main olfactory bulb (MOB) neuronal spontaneous activity were investigated using extracellular recordings in mouse in vivo. These effects were compared with those of another nociceptive supplement, the μ-opioid agonist buprenorphine. Ketoprofen (100 or 200 mg/kg) did not significantly alter MOB single-unit spontaneous rates in either ICR or C57BL/6J mice. In contrast, buprenorphine, at doses of 0.02, 0.05, and 0.20 mg/kg, inhibited MOB neuronal spontaneous rates by 19%, 49%, and 57%, respectively. Neither drug altered the temporal patterning of single-unit spike trains, as measured by the interspike interval (ISI) coefficient of variation (CV). We also investigated the ability of ketoprofen and buprenorphine to induce antinociception in the anesthetized mouse. The electroencephalogram (EEG) was used to measure the anesthetic plane. Both ketoprofen and buprenorphine altered the EEG trace and ketoprofen altered the power spectrum in a manner consistent with deepening anesthesia. Lastly, when applied at the time of anesthesia induction, ketoprofen decreased the amount of chloral hydrate necessary to maintain a defined anesthetic plane during the rest of the experiment. These results suggest that ketoprofen induces antinociception under chloral hydrate anesthesia without significantly inhibiting spontaneous activity of MOB neurons. Ketoprofen is therefore suitable as an antinociceptive supplement to chloral hydrate anesthesia during in vivo electrophysiologic recordings of the mouse MOB.

Optical coherence tomography (OCT) and laser-induced fluorescence (LIF) spectroscopy each have clinical potential in identifying human gastrointestinal (GI) pathologies, yet their diagnostic capability in mouse models is unknown. In this study, we combined the 2 modalities to survey the GI tract of a variety of mouse strains and ages and to sample dysplasias and inflammatory bowel disease (IBD) of the intestines. Segments (length, 2.5 cm) of duodenum and lower colon and the entire esophagus were imaged ex-vivo with combined OCT and LIF. We evaluated 30 normal mice (A/J and 10- and 21-wk-old and retired breeder C57BL/6J) and 10 mice each of 2 strains modeling colon cancer and IBD (Apc^Min and IL2-deficient mice, respectively). Histology was used to classify tissue regions as normal, Peyer patch, dysplasia, adenoma, or IBD. Features in corresponding OCT images were analyzed. Spectra from each category were averaged and compared via Student t tests. OCT provided structural information that led to identification of the imaging characteristics of healthy mouse GI. With histology as the 'gold standard,' we developed preliminary image criteria for early disease in the form of adenomas, dysplasias, and IBD. LIF characterized the endogenous fluorescence of mouse GI tract, with spectral features corresponding to collagen, NADH, and hemoglobin. In the IBD sample, LIF emission spectra displayed potentially diagnostic peaks at 635 and 670 nm, which we attributed to increased porphyrin production by bacteria associated with IBD. OCT and LIF appear to be useful and complementary modalities for ex vivo imaging of mouse GI tissues.

Assisted ventilation is necessary for treating preterm infants with respiratory distress syndrome. Unfortunately, high and prolonged concentrations of oxygen associated with assisted ventilation often lead to pulmonary changes, such as hemorrhage and inflammation. The resulting chronic pulmonary condition is known as bronchopulmonary dysplasia. Pulmonary changes characteristic of this syndrome can be produced in rat pups exposed to high oxygen levels. We exposed 21-d-old rats to room air or continuous 95% oxygen for 7 d and then allocated them into 6 groups to evaluate whether treatment with zileuton and zafirlukast, 2 agents which decrease the effects of leukotrienes, lessened the pulmonary effects of short-term hyperoxia. After 7 d, lung tissue was collected for light and electron microscopy. Pulmonary changes including edema, hemorrhage, alveolar macrophage influx, and Type II pneumocyte proliferation were graded on a numerical scoring system. Compared with controls exposed to hypoxia and saline, rats exposed to hypoxia and treated with zileuton had significantly reduced levels of alveolar macrophage influx and Type II pneumocyte proliferation, but those exposed to hypoxia and treated with zafirlukast showed no significant reduction in any pulmonary changes. This study helps define pulmonary changes induced secondary to hyperoxia in rat pups and presents new information on the mechanisms of leukotriene inhibition in decreasing the severity of hyperoxic lung injury.

The SPRD-Pkdr1 rat model is widely used for the study of human autosomal dominant polycystic kidney disease. This rat model carries the Cy allele of the Pkdr1 gene, which results in polycystic kidney disease. Because the Cy allele is lethal in the homozygous state at weanling age, the breeding colony must be maintained in the heterozygous state. A random breeding scheme in which production of homozygous pups with enlarged kidneys indicates heterozygous breeders is commonly used. This study was performed to determine whether biochemical markers (blood urea nitrogen [BUN] or creatinine), ultrasonography, or genetic analysis could be used to select breeding animals in the SPRD-Pkdr1/Rrrc colony and thus replace the random breeding scheme with a more efficient selective breeding scheme. BUN was predictive of the Cy allele in 8- to 9-wk-old male but not female rats. Ultrasonography identified animals with polycystic kidney disease in both sexes by 9 wk of age. Microsatellite marker polymorphism analysis could not be used to determine carrier status for the Cy allele, but restriction fragment length polymorphism analysis appropriately detected the Cy allele in 100% of the animals examined. In conclusion, multiple methods can be used for detecting the Cy allele, making possible a selective breeding scheme that markedly reduces the necessary number of breeder animals and eliminates the euthanasia of offspring needed with a random test-mating scheme.

Ischemic preconditioning (IPC) not only reduces local tissue injury caused by subsequent ischemia-reperfusion (IR) but may also have a beneficial effect on IR injury of tissues remote from those undergoing preconditioning. In this study, we investigated the effect of small intestinal IPC on renal IR injury in rats. Renal IR injury was induced by a 45-min renal artery occlusion and reperfusion for 2 or 24 h in rats with a previous contralateral nephrectomy, and ischemic preconditioning was induced by 3 cycles of 8-min ischemia and 5-min reperfusion of the small intestine. We then measured the concentrations of plasma creatinine (Cr) and blood urine nitrogen (BUN) and the level of malondialdehyde (MDA) and activities of superoxide dismutase (SOD) and catalase (CAT) in the renal cortex. Renal histopathology also was evaluated. Pretreatment with intestinal ischemic preconditioning significantly alleviated renal IR injury, as shown by decreases in the levels of Cr, BUN, and MDA, decreased renal morphologic change, and improved preservation of SOD and CAT activities. These results suggest that remote ischemic preconditioning of the small intestine protects against renal IR injury by inhibition of lipid peroxidation and preservation of antioxidant enzyme activities.

Trypanoxyuris microon is a pinworm that infects New World nonhuman primates, including Aotus nancymae. Although it typically is clinically insignificant, infection may serve as a significant variable during experimental data analysis. In this study we sought to determine the most effective anthelmintic therapy for eradication of T. microon infection in A. nancymae. Animals confirmed to be infected with T. microon by perianal tape test were treated twice (on days 0 and 14) with pyrantel pamoate, ivermectin, or thiabendazole and evaluated for eggs by daily perianal tape test throughout the entire 28-d period. Successful clearance of eggs was defined as 5 consecutive negative perianal tape tests. Pyrantel pamoate and ivermectin were significantly more effective at egg clearance than were thiabendazole and no treatment. Overall, 100% of the pyrantel pamoate and ivermectin treatment groups were cleared of infection after 2 treatments, whereas only 60% of the thiabendazole group became negative for pinworm eggs. In addition, the time after treatment until clearance was 1 to 2 d for pyrantel pamoate, 2 to 4 d for thiabendazole, and 4 to 6.5 d for ivermectin. These results indicate that pyrantel pamoate was the most effective and rapidly acting anthelmintic for the treatment of adult T. microon infection, with ivermectin as a suitable alternative. However because of the potential for continued development of immature stages or reinfection, anthelmintic doses should be repeated after 1 to 2 wk, in combination with effective environmental sanitation.

Under specific pathogen-free conditions, 1.3% to 1.8% of litters born in our inbred 101/H and C3HeB/FeJ mouse colonies had pups with steatorrhea and runting. Clinically affected male and female pups were first identified when they were from 14 to 25 d old. Unaffected littermates were healthy and were weaned successfully. Postmortem findings in 8 clinically affected mice included a small, poorly differentiated exocrine pancreas comprising cytokeratin-negative duct-like structures but lacking recognizable acinar cells with their normal carboxypeptidase B-positive zymogen granules. Endocrine pancreas islets were unremarkable and contained insulin-positive β cells and glucagon-positive α cells. There was mild inflammation of the hindgut but no evidence of intestinal pathogens or marked inflammation or necrosis of pancreas, either alone or as part of a multisystemic inflammatory condition. Sera from pups in 4 affected litters did not contain antibodies to reovirus 3, mouse coronavirus, rotavirus, or mouse adenovirus 2. Furthermore, 4 sets of parental mice and sentinel mice from the facility were negative for 13 viruses, bacteria, and parasites. C3HeB/FeJ and 101/H inbred strains may be genetically predisposed because the steatorrhea and runting was absent in 13 other mouse strains and subspecies bred in the specific pathogen-free facility. This condition resembles exocrine pancreas hypoplasia, but the inheritance is complex. A wider implication is that runting coupled with steatorrhea are phenotypic criteria to suspect pancreatic disease that could be used in the context of a mouse N-ethyl-N-nitrosourea-mutagenesis program to identify potential mutants with defects in pancreas development.