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Routine health monitoring is an integral part of managing SPF rodent colonies. In recent years, rack-level environmental sampling has been introduced as an adjunct method or replacement for exposure of sentinel rodents to soiled bedding. However, rack-level environmental monitoring is not compatible with rodent housing systems that have cage-level filtration. The current study investigated whether exposure of sterile flocked swabs to soiled bedding can be an alternative sampling method for routine health monitoring in mice, thus replacing the use of sentinels in soiled-bedding cages. Flocked swabs were placed in cages containing pooled samples of soiled bedding but no mice; swabs remained there for 90 d, with weekly agitation and biweekly swabbing of the cage floor to mimic the agitation of soiled bedding by sentinel mice and facilitate the collection of dust particles. Fecal samples were collected from both colony and sentinel mice. For environmental samples, exhaust debris was collected from the rack plenum, and dust samples were collected from the exhaust hose. All samples were collected on days 88 through 91 and were tested for multiple pathogens by using real-time PCR assays. To determine the diagnostic agreement of flocked swab sampling with the other methods, we used κ statistics to compare the test results from flocked swabs with those from sentinel feces, exhaust debris, and colony animal feces; we found excellent agreement between the colony feces and the flocked swab methods. The sterile flocked swab method detected all enzootic pathogens in the colonies tested. Results from flocked swab samples had the least agreement with sentinel feces, which also failed to detect the presence of fur mites. This study supports the use of sterile flocked swabs as alternative to using sentinel mice, thus conforming to the guiding principles of replacement and reduction in the use of animals for routine colony health monitoring.

China, Japan, and Korea have spent decades developing and amending laws, regulations, and guidelines to address the humane care and use of laboratory animals. This process began in 1983 in China, 1973 in Japan, and 1991 in Korea and has continued to the present. The governmental oversight of research varies between these countries, ranging from regulations by multiple levels of government in China to self-regulation under multiple government guidelines in Japan. Common to all is incorporation of the internationally recognized principles of the 3Rs: replacement, reduction and refinement. This paper reviews how the framework of laws, regulations, and guidelines evolved in each of these countries, their current status, and the expectation that they will continue to evolve.

It is well recognized that animals play a vital role and are indispensable to scientific and medical research. Over the years, a number of non-animal procedures have been developed. However, despite all the advances in science, as yet, no system has been evolved which can completely replace a living system to conduct basic research. There is still a need to test food, drugs, medical devices, treatment regimes etc. on some animals before they can be tested and used (if found suitable) in human beings. Even the most sophisticated technology models have failed to mimic completely the complex cellular interactions occurring in a living system. The search for a complete alternative to animal research is still on and in the mean time we can all help play our part by conducting animal research in a humane and responsible fashion. This chapter discusses the ethical issues in animal research highlighting the need to use animals conscientiously.

To identify an appropriate model of human renin-angiotensin system (RAS) involvement in fetal origins of adult disease, we quantitated renal ANG II AT1 and AT2 receptors (AT1R and AT2R, respectively) in fetal (90-day gestation, n = 14), neonatal (3-wk, n = 5), and adult (6-mo, n = 8) microswine by autoradiography (125I-labeled [Sar1Ile8]ANG II+cold CGP-42112 for AT1R,125I-CGP-42112 for AT2R) and by whole kidney radioligand binding. The developmental pattern of renal AT1R in microswine, like many species, exhibited a 10-fold increase postnatally ( P < 0.001), with maximal postnatal density in glomeruli and lower density AT1R in extraglomerular cortical and outer medullary sites. With aging, postnatal AT1R glomerular profiles increased in size ( P < 0.001) and fractional area occupied ( P < 0.04), with no change in the number per unit area. Cortical levels of AT2R by autoradiography fell with age from ≅5,000 fmol/g in fetal kidneys to ≅60 and 20% of fetal levels in neonatal and adult cortex, respectively ( P < 0.0001). The pattern of AT2R binding in postnatal pig kidney mimicked that described in human and simian, but not rodent, species: dense AT2R confined to discrete cortical structures, including pre- and juxtaglomerular, but not intraglomerular, vasculature. Our results provide a quantitative assessment of ANG II receptors in developing pig kidney and document the concordance of pigs and primates in developmental regulation of renal AT1R and AT2R.

Objective — To identify vascular cells capable of responding to angiotensin II (Ang II) generated in conduit arteries, we examined the Ang II type 1 receptor (AT1R) and Ang II type 2 receptor (AT2R) in the thoracic aorta (TA) and abdominal aorta (AA) and branches in 90-day fetal, 3-week postnatal, and 6-month adult microswine. Methods and Results — By autoradiography ( 125 I-[Sar 1 Ile 8 ]-Ang II with or without AT1R- or AT2R-selective analogues or 125 I - CGP 42112), there were striking rostrocaudal differences in (1) AT2R binding at all ages (prominent in AA wall and branches, sparse in TA wall and branches) and (2) a non-AT2R binding site for CGP 42112 (consistently evident in postnatal TA and branches but absent in AA and branches). Furthermore, patterns of AT2R distribution in infradiaphragmatic arteries were developmentally distinct. In fetal AAs, high-density AT2Rs occupied the inner 60% of the medial-endothelial wall. In postnatal AAs, AT2Rs were sparse in the medial-endothelial wall but prominent in a circumferential smooth muscle α-actin–negative cell layer at the medial-adventitial border, occupying ≈20% to 25% of the AA cross-sectional area. AT1R density in the TA and AA medial-endothelial wall increased with age, whereas AT2R density decreased after birth. Conclusions — A novel AT2R-positive cell layer confined to postnatal infradiaphragmatic arteries physically links adventitial and medial layers, appears optimally positioned to transduce AT2R-dependent functions of local Ang II, and suggests that adventitial Ang II may elicit regionally distinct vascular responses.

Octreotide, a somatostatin analog, was evaluated for its effects on long-term survival in a mouse model of endotoxemia and for its effects on endotoxin-induced suppression of human leukocyte migration. Swiss Webster mice were simultaneously rendered endotoxemic with a single intraperitoneal injection of 800 micrograms E. coli Lipopolysaccharide (LPS) and treated with one of four doses of subcutaneous (s.c.) octreotide (1.0 mg/kg in 0.4 ml saline, 0.1 mg/kg in 0.4 ml saline, 0.1 mg/kg in 0.04 ml saline, or 0.001 mg/kg in 0.04 ml saline) or saline alone (fluid-resuscitated control group: 0.4 ml saline s.c.; or non-fluid-resuscitated control group: 0.04 ml saline s.c.). Octreotide was continued with or without supplemental s.c. fluid resuscitation (0.4 ml saline) at eight hour intervals for either twenty-four or forty hours. There was no statistical significance to differences in long-term survival between comparable groups of octreotide treated vs saline treated animals during the entire fourteen day period of observation. Fluid resuscitation during the first forty hours following endotoxemia induction delayed death, but did not significantly improve long-term survival. In vitro work was conducted to determine the effect of octreotide on endotoxin-induced suppression of human leukocyte migration. Octreotide at concentrations ranging from 3.05 x 10(-5) Molar to 3.05 x 10(-11) Molar had no significant effect on leukocyte migration. In this study octreotide treatment failed to improve long-term survival in mice with endotoxemia and did not alter endotoxin-induced suppression of leukocyte migration.

Pregnant animals are less able to maintain mean arterial pressure (MAP) during hemorrhage compared with nonpregnant animals, but the hemodynamic basis of this difference is unknown. The hypothesis that pregnancy attenuates responses of cardiac output, as well as total peripheral resistance (TPR) and femoral conductance, to hemorrhage was tested in conscious rabbits in both the pregnant and nonpregnant state ( n = 10). During continuous slow blood loss (2% of the initial blood volume per minute), MAP was maintained initially in both groups. However, MAP then abruptly decreased to