Cardiac phenotyping of transgenic mice typically requires anesthesia. Chemical-grade tribromoethanol (TBE) is commonly used for this purpose due to its relatively short duration of action, modest cardiodepressive effects, and its noncontrolled status. In the present study, we used both genders of C57BL/6;C3H-Tg(Slc8a1)hKdp transgenic (TG) mice and C57BL/6;C3H wild-type (WT) mice to evaluate isoflurane (ISF) as a pharmaceutical-grade alternative to TBE for echocardiography and electrocardiography. Baseline target physiologic heart rates (beats per minute) were established by use of telemetry as 544 ± 10 in WT mice and 580 ± 21 in TG mice. TG and WT animals were anesthetized with either 0.8% to 1% inhalational ISF or 250 mg/kg intraperitoneal TBE. The following parameters were measured or calculated according to the previously defined physiologic heart rates: end diastolic and systolic dimensions; posterior wall and ventricular septal thicknesses; left ventricular mass, aortic ejection times; left ventricular fractional shortening; velocity of circumferential fiber shortening; and left ventricular ejection fraction. No significant difference between anesthetics was found for any measured cardiac parameters. However, the time required for data acquisition was significantly shorter for ISF (10 min) than for TBE (14 min). This study demonstrates that comparable echocardiographic results can be obtained at higher throughput by use of pharmaceutical-grade ISF than with chemical-grade TBE.

Microbial infections are common sequelae in humans and animals implanted with long-term intravascular catheters. Understanding the pathophysiology of infectious morbidity is critical to improving quality of care in catheterized subjects. Here, we describe findings in 6 clinically healthy, male sheep implanted with indwelling aortic or cardiac catheters for 6 to 10 mo. We isolated multiple bacterial species including Serratia spp., Enterobacter agglomerans, Eschericia coli, Klebsiella oxytoca, and K. pneumoniae in aerobic cultures from catheter tips. Although sheep were clinically asymptomatic, 1 or both kidneys from all animals contained wedge-shaped infarcts of varying size and number. Microscopic examination revealed (a) marked fibrosis with mild inflammatory cell infiltrate consistent with chronic foreign body reaction around catheters; (b) moderate to severe, diffuse, subacute to chronic membranoproliferative glomerulonephritis and mild, multifocal chronic interstitial nephritis; and (c) mesangial immune-complex deposition as demonstrated by direct immunofluorescence technique. The finding of bacterial colonization of catheters together with chronic glomerulonephritis and immune-complex deposits in kidneys in clinically asymptomatic sheep underscores the need for close microbiologic monitoring of catheter implants and assessment of kidney function in animals instrumented for long-term vascular access.

The soy isoflavones genistein and daidzein, as well as the daidzein metabolite equol, have structural similarities to mammalian estrogens and bind with varying affinity to both known subtypes of the estrogen receptor. Consequently, prospective studies in both humans and animals have begun to evaluate the potential effects of isoflavones on estrogen receptor-mediated phenomena. However, many diets of laboratory-housed animals derive their protein from soy and thus likely contain substantial quantities of isoflavones. Exposing experimental subjects to these isoflavones via such diets could confound studies, particularly those evaluating the effects of estrogen or estrogen-like ligands. The aim of this study was to compare the levels of circulating concentrations of isoflavones and their metabolites in monkeys fed either a soy-free diet, a soy-based diet providing 130 mg of isoflavone (daidzein, genistein, and glycitein aglycon equivalents) daily, or a commercially available 'chow' diet containing an unspecified amount of soybean meal. Animals consuming the commercial diet had serum concentrations of daidzein, genistein, and glycitein that were significantly higher than those of animals fed a soy-free diet but similar to those of monkeys fed a soy-based diet formulated to be high in isoflavones. Notably, animals fed the commercial diet also had serum equol concentrations that were similar to or, in some cases, in excess of serum concentrations in the animals fed the soy diet. These data argue for the use of soy-free diets in studies investigating estrogenic effects on physiologic or behavioral endpoints.

Timely and accurate detection of murine pathogens is essential in contemporary biomedical research. Cost, accuracy, and reproducibility of test results are frequent concerns when initiating an on-site serology program. This study was conducted to evaluate the advantages of on-site serology performed by enzyme-linked immunosorbent assay (ELISA) versus pathogen surveillance conducted off-site by a commercial vendor. We divided 92 sentinel mouse serum samples and tested them in parallel for a panel of 10 murine pathogens at our institution and by an off-site vendor. On-site testing was performed with commercially available test kits and according to the kit manufacturer's directions, whereas serum samples for off-site testing were prepared according to the vendor's specifications. Results from the 2 testing strategies were compared, and a good beyond-chance level of agreement was demonstrated by means of the kappa test (κ = 0.86). The turn-around time between sample preparation and results availability for on-site ELISA was 16 h versus 72 h for off-site testing. On-site ELISA demonstrated considerable cost reduction, ranging from 15.10% to 43.33% depending on the number of agents being tested. This study demonstrates the accuracy and time- and cost-effectiveness of on-site ELISA as well as its potentially valuable role in achieving more timely and efficient disease surveillance and control programs in contemporary biomedical research facilities.

Ectoparasites pose numerous research, health, and management problems for researchers and institutions. Our facility management experience was complicated by recurrence of murine fur mite (Radfordia affinis) infestation after several rounds of single-mode fur mite treatment with dichlorvos in the cage bedding. Subsequently, we successfully eradicated the fur mites using a multidrug therapeutic protocol. Over an 8-wk treatment period, 2 applications of topical selamectin were administered in conjunction with amitraz- and fipronil-treated nestlets changed weekly. Mice tolerated the therapy well with no side effects noted, and to date there has been no recrudescence. To our knowledge, this report is the first to describe combined use of these specific therapeutic agents to control fur mite infestation in laboratory mice.

Preparation for the specialty board examination for the American College of Laboratory Animal Medicine (ACLAM) is an intensive process that is facilitated by geographic regions where many people studying for the exam are located in close proximity. However, many people work at institutions that are distant from these 'study centers.' Approximately 10 y ago, the Laboratory Animal Boards Study Group (LABSG) online journal club was established to provide a forum for journal review for examination preparation. Over the years, the mission of this group has expanded to include practice examinations and practicals, questions from common resources, and summaries and questions from common laboratory animal science journals. These study aids are beneficial for those preparing for the ACLAM certification examination. They are also beneficial for those preparing for the technician and manager certification examinations offered by the American Association for Laboratory Animal Science (AALAS). This article is intended to be an introduction to the variety of study aids available through the LABSG online journal review club and the LABSG web page (www.labsg.org). It also provides details on the demographics of participants and an exploration of how this resource enhances examination preparation.

We developed a surgical procedure for accessing the prostate gland of the cynomolgus monkey (Macaca fascicularis) through the perineal cavity. The procedure can be used for direct injection of compounds into the prostate gland and (or) for the collection of biopsies. The rationale for developing this technique at our site was the need for precise injection into the gland with a low probability of error, as the compound tested in a subsequent study required prostate-specific antigen for activation. A perianal incision was made approximately 1 cm ventral to the anus, and the muscle and subcutaneous tissue were bluntly dissected between the urethra and the rectum. The prostate gland was easily visualized after dissection, and could be grasped gently by the capsule and exteriorized through the incision, thus allowing easy access to the prostate for study purposes. On the basis of mock injections with methylene blue dye and gross observation of prostate tissue at necropsies immediately after injection, we recommend that 2 injections be given per lobe of prostate, and injections should be to a depth of 2 to 3 mm to provide uniform distribution of injected compounds. To minimize back pressure and leakage from the injection site, a small-gauge needle (23-27 gauge) should be used and the needle held in place for approximately 30 s before withdrawal. Injection volumes 64 μl per g prostate or less did not cause the back ow of methylene blue dye into the seminal vesicles.

Rising concerns over respiratory illnesses caused by agents such as avian influenza viruses and SARS coronavirus have prompted intensive research efforts and the resurgence of nonhuman primates as models for these human diseases. In the context of studying influenza infection and vaccine development, serial bronchoscopic procedures, including bronchial brush biopsies and bronchoalveolar lavage, were performed in pigtailed macaques (Macaca nemestrina). The possible need for oxygen supplementation during these procedures was anticipated because of the size of the animals relative to the 5-mm bronchoscope. We therefore monitored oxyhemoglobin saturation, a measure of arterial blood oxygen content, before and after insertion of the bronchoscope, during bronchoalveolar lavage, and after initiation of oxygen supplementation. Although more data are required to draw definitive conclusions, our findings suggested the need for oxygen supplementation during such procedures in nonhuman primates, despite the fact that human patients undergoing bronchoscopy and lavage do not routinely get oxygen unless they are already compromised. Our data also suggested that the need for supplementation could not be predicted from simple parameters such as size of the animal, presence of respiratory clinical signs, or experimental treatment. Finally, we show a simple and cost-effective method of using human nasal cannulas for delivering oxygen to pigtailed macaques during bronchoscopic procedures, and we believe that, after further testing, this method could be used safely and effectively in other nonhuman primate species.

Here we present an echographic method to withdraw amniotic fluid from pregnant rats. The method could be an alternative to the surgical amniotic fluid collection methods used currently. On day 18 of gestation, pregnant Sprague–Dawley rats underwent amniotic sac puncture by either surgical procedure or echographically guided method. This study evaluated the effect of the 2 collection procedures on parturition day, number of pups per litter, and weight of newborns compared with those of a control group without any fluid collection. These parameters did not differ statistically across groups. However, the echographically guided method did not require surgery or postsurgical recovery and was therefore advantageous from the perspective of animal use. Moreover ultrasound-guided collection allows experimental designs that require collection of multiple samples from the same animal during a single pregnancy.

The increased use in noninvasive imaging of laboratory rodents has prompted innovative techniques in animal handling. Lung imaging of rodents can be a difficult task because of tissue motion caused by breathing, which affects image quality. The use of a prototype flat-panel computed tomography unit allows the acquisition of images in as little as 2, 4, or 8 s. This short acquisition time has allowed us to improve the image quality of this instrument by performing a breath-hold during image acquisition. We designed an inexpensive and safe method for performing a constant-pressure breath-hold in intubated rodents. Initially a prototypic manual 3-way valve system, consisting of a 3-way valve, an air pressure regulator, and a manometer, was used to manually toggle between the ventilator and the constant-pressure breath-hold equipment. The success of the manual 3-way valve system prompted the design of an electronically actuated valve system. In the electronic system, the manual 3-way valve was replaced with a custom designed 3-way valve operated by an electrical solenoid. The electrical solenoid is triggered by using a hand-held push button or a foot pedal that is several feet away from the gantry of the scanner. This system has provided improved image quality and is safe for the animals, easy to use, and reliable.

Injectable anesthetic drugs used in rodents are often mixed and further diluted to increase the convenience and accuracy of dosing. We evaluated clinical refractometry as a simple and rapid method of quality control and mixing error detection of rodent anesthetic or analgesic mixtures. Dilutions of ketamine, xylazine, acepromazine, and buprenorphine were prepared with reagent-grade water to produce at least 4 concentration levels. The refraction of each concentration then was measured with a clinical refractometer and plotted against the percentage of stock concentration. The resulting graphs were linear and could be used to determine the concentration of single-drug dilutions or to predict the refraction of drug mixtures. We conclude that refractometry can be used to assess the concentration of dilutions of single drugs and can verify the mixing accuracy of drug combinations when the components of the mixture are known and fall within the detection range of the instrument.

Urolithiasis and carcinoma of the urinary bladder are generally uncommon in rats. In particular, spontaneous urolithiasis and carcinoma of the bladder in young rats has not yet been reported. Our case concerns the unexpected finding of urolithiasis of the bladder and left kidney of an apparently healthy 4-mo-old male Wistar rat. The bladder was filled with 28 white, round to oval calculi of various sizes. The kidney presented a single unevenly shaped calculus in the pelvis. Histology revealed, in addition to urolithiasis, a grade II papillary transitional cell carcinoma of the bladder, with muscle invasion and neoplastic extension to the left renal pelvic epithelium. No previous experimental procedure or dietetic manipulation had occurred, which could be held responsible for these findings. These unusual findings indicate that, unknown to researchers, unapparent factors can affect the health status of even relatively young experimental animals. Asymptomatic disease, such as the unknown presence of a tumor, potentially can affect many parameters and thus influence research results.