Comparison of Antemortem and Environmental Samples for Zebrafish Health Monitoring and Quarantine.

Molecular diagnostic assays offer both exquisite sensitivity and the ability to test a wide variety of sample types. Various types of environmental sample, such as detritus and concentrated water, might provide a useful adjunct to sentinels in routine zebrafish health monitoring. Similarly, antemortem sampling would be advantageous for expediting zebrafish quarantine, without euthanasia of valuable fish.

Influence of Rack Design and Disease Prevalence on Detection of Rodent Pathogens in Exhaust Debris Samples from Individually Ventilated Caging Systems.

Sampling of bedding debris within the exhaust systems of ventilated racks may be a mechanism for detecting murine pathogens in colony animals. This study examined the effectiveness of detecting pathogens by PCR analysis of exhaust debris samples collected from ventilated racks of 2 different rack designs, one with unfiltered air flow from within the cage to the air-exhaust pathway, and the other had a filter between the cage and the air-exhaust pathway. For 12 wk, racks were populated with either 1 or 5 cages of mice (3 mice per cage) infected with one of the following pathogens: mouse norovirus (MNV), mouse parvovirus (MPV), mouse hepatitis virus (MHV), Helicobacter spp.

Development of a PCR assay for the detection of Spironucleus muris.

Spironucleus muris is a protozoan that can colonize the intestinal tract of many rodent species. Although its effects on animal health and research are debated, S. muris is often included on exclusion lists for rodent facilities.

Development of a virus concentration method using lanthanum-based chemical flocculation coupled with modified membrane filtration procedures.

Direct membrane filtration is often used to concentrate viruses in water but it may suffer from severe membrane fouling and clogging. Here, a lanthanum-based flocculation method coupled with modified membrane filtration procedures was developed and evaluated to detect viruses in large volume (40 L) water samples. The lanthanum-based flocculation method could easily reduce the water sample volume by a factor of 40.

Viral diseases in zebrafish: what is known and unknown.

Naturally occurring viral infections have the potential to introduce confounding variability that leads to invalid and misinterpreted data. Whereas the viral diseases of research rodents are well characterized and closely monitored, no naturally occurring viral infections have been characterized for the laboratory zebrafish (Danio rerio), an increasingly important biomedical research model. Despite the ignorance about naturally occurring zebrafish viruses, zebrafish models are rapidly expanding in areas of biomedical research where the confounding effects of unknown infectious agents present a serious concern.

Completion of the swine genome will simplify the production of swine as a large animal biomedical model.

Background: Anatomic and physiological similarities to the human make swine an excellent large animal model for human health and disease.

Methods: Cloning from a modified somatic cell, which can be determined in cells prior to making the animal, is the only method available for the production of targeted modifications in swine.

Results: Since some strains of swine are similar in size to humans, technologies that have been developed for swine can be readily adapted to humans and vice versa.

Determination of low-density Escherichia coli and Helicobacter pylori suspensions in water.

Determination of low-density of bacteria, especially those of pathogenic strains in water, has proven difficult and challenging. Here, we developed and evaluated a lanthanum-based concentration method coupled with quantitative real-time PCR to concentrate and detect selected bacteria (Escherichia coli and Helicobacter pylori) in water. To improve qPCR efficiency, the flocs with enmeshed bacteria after chemical flocculation need to be dissolved before PCR detection.

Pneumocystis carinii infection causes lung lesions historically attributed to rat respiratory virus.

Idiopathic lung lesions characterized by dense perivascular cuffs of lymphocytes and a lymphohistiocytic interstitial pneumonia have been noted in research rats since the 1990s. Although the etiology of this disease has remained elusive, a putative viral etiology was suspected and the term 'rat respiratory virus' (RRV) has been used in reference to this disease agent. The purpose of this study was to determine whether Pneumocystis carinii infection in immunocompetent rats can cause idiopathic lung lesions previously attributed to RRV.

Detecting food- and waterborne viruses by surface-enhanced Raman spectroscopy.

Food- and waterborne viruses pose serious health risks to humans and were associated with many outbreaks worldwide. Rapid, accurate, and nondestructive methods for detection of viruses are of great importance to protect public health. In this study, surface-enhanced Raman spectroscopy (SERS) coupled with gold SERS-active substrates was used to detect and discriminate 7 food- and waterborne viruses, including norovirus, adenovirus, parvovirus, rotavirus, coronavirus, paramyxovirus, and herpersvirus.

Evaluation of a commercial colorimetric fecal dipstick assay for the detection of Helicobacter hepaticus infections in laboratory mice.

Mice used in biomedical research typically are tested for the presence of Helicobacter spp., including Helicobacter hepaticus. Here we evaluated the ability of a commercially available colorimetric Helicobacter dipstick assay to detect H.

Lanthanum-based concentration and microrespirometric detection of microbes in water.

Rapid concentration and detection of microorganisms, particularly pathogens, are important but remain a challenge. In this research, lanthanum chloride (LaCl(3)) was used to concentrate E. coli in water and the results were compared with those obtained using traditional flocculants, such as ferric sulfate and aluminum sulfate.

Workshop summary: detection, impact, and control of specific pathogens in animal resource facilities.

Despite advances, infectious diseases remain a threat to animal facilities, continue to affect animal health, and serve as potential confounders of experimental research. A workshop entitled Detection, Impact, and Control of Specific Pathogens in Animal Resource Facilities was sponsored by the National Center for Research Resources (NCRR) and National Institutes of Aging (NIA) and held April 23-24, 2009, at the Lister Hill Conference Center on the National Institutes of Health's (NIH) Bethesda campus. The meeting brought together laboratory animal scientists and veterinarians with experience in fish, rodent, and nonhuman primate models to identify common issues and problems.

Mutational insertion of a ROSA26-EGFP transgene leads to defects in spermiogenesis and male infertility in mice.

Pronuclear injection has been a successful strategy for generating genetically engineered mouse models to better understand the functionality of genes. A characteristic of pronuclear injection is that random integration of the transgene into the genome can disturb a functional gene and result in a phenotype unrelated to the transgene itself. In this study, we have characterized a mouse model containing an insertional mutation that, in the homozygous state, severely affects spermatogenesis as characterized by lack of sperm motility and acrosomal aplasia.

Proceedings of the Conference on Swine in Biomedical Research.

The Conference on Swine in Biomedical Research was held April 2-3, 2008, in San Diego, California. The goal of the conference was to bring together individuals who are using swine as models, creating new swine models, or studying human health and disease. This is the only conference that focuses exclusively on swine models and as such is the premier meeting for investigators who use or develop swine as models for biomedical research.

Differential susceptibility of SD and CD rats to a novel rat theilovirus.

Antibodies to rat theilovirus (RTV) have been detected in rats for many years because of their serologic crossreactivity with strains of Theiler murine encephalomyelitis virus (TMEV) of mice. Little information exists regarding this pathogen, yet it is among the most common viruses detected in serologic surveys of rats used in research. In the study reported here, a novel isolate of RTV, designated RTV1, was cultured from the feces of infected rats.

Lurking in the shadows: emerging rodent infectious diseases.

Rodent parvoviruses, Helicobacter spp., murine norovirus, and several other previously unknown infectious agents have emerged in laboratory rodents relatively recently. These agents have been discovered serendipitously or through active investigation of atypical serology results, cell culture contamination, unexpected histopathology, or previously unrecognized clinical disease syndromes.

Multiplex microsatellite marker panels for genetic monitoring of common rat strains.

Because microsatellite markers have a high degree of genetic variability, they are an effective tool for genetic monitoring. We have developed a genotyping panel containing 87 microsatellite markers that are polymorphic among commonly used inbred rat strains, including ACI, Fischer 344, Lewis, Brown Norway, Wistar-Furth, and Wistar-Kyoto. The markers are located at approximately 15- to 20-cM intervals along each of the 20 autosomes.

Soiled-bedding sentinel detection of murine norovirus 4.

According to serologic surveys, murine norovirus (MNV) is the most prevalent viral pathogen infecting mice used in biomedical research. However, the use of sentinel mice to detect MNV-infected mouse populations has not been evaluated thoroughly. To this end, an experimental method of soiled bedding transfer was created to mimic a quarterly sentinel monitoring program.

Progress and prospects in rat genetics: a community view.

The rat is an important system for modeling human disease. Four years ago, the rich 150-year history of rat research was transformed by the sequencing of the rat genome, ushering in an era of exceptional opportunity for identifying genes and pathways underlying disease phenotypes. Genome-wide association studies in human populations have recently provided a direct approach for finding robust genetic associations in common diseases, but identifying the precise genes and their mechanisms of action remains problematic.

Multiplex fluorescent immunoassay for the simultaneous detection of serum antibodies to multiple rodent pathogens.

Multiplex Fluorescent Immunoassay (MFI) is a sensitive and specific serologic test that allows simultaneous detection of antibodies to multiple viral and bacterial agents in a single reaction well. MFI is a high-throughput assay that offers several advantages over other prevalent assays, and some research animal diagnostic laboratories have adopted it as their primary technique. The authors present a detailed review of MFI and its application to laboratory animal diagnostics.

Detection of mouse parvovirus in Mus musculus gametes, embryos, and ovarian tissues by polymerase chain reaction assay.

We used primary and nested polymerase chain reaction (PCR) assays to determine the presence of mouse parvovirus (MPV) in mouse sperm, oocytes, preimplantation embryos, and ovarian tissues collected from MPV-infected mice. The primary PCR assay detected MPV in 56% of the sperm samples. MPV was not eliminated by passing sperm samples through a Percoll gradient.

Molecular characterization of three novel murine noroviruses.

Murine noroviruses (MNV) comprise a group of newly recognized pathogens infecting laboratory mice. The first reported murine norovirus, murine norovirus 1 (MNV-1), produces a transient infection with a short duration of fecal shedding after infection of immunocompetent laboratory mice. Our laboratory subsequently isolated three novel murine noroviruses, murine norovirus 2 (MNV-2), murine norovirus 3 (MNV-3), and murine norovirus 4 (MNV-4), that have markedly different pathogenicity from MNV-1 by producing persistent infections and prolonged fecal shedding in infected immunocompetent mice.

Persistent infection with and serologic cross-reactivity of three novel murine noroviruses.

According to serologic testing, murine norovirus is the most prevalent viral pathogen in contemporary laboratory mice. Previously, murine norovirus 1 (MNV-1) was the only norovirus reported to infect research mice. In this study, 3 novel murine norovirus strains--MNV-2, MNV-3, and MNV-4--were isolated from geographically separate mouse research colonies.

Improved survival of vitrified porcine embryos after partial delipation through chemically stimulated lipolysis and inhibition of apoptosis.

Mechanical removal of intracellular lipids has been the most effective approach to increase the cryosurvival of porcine embryos. In this experiment, we tested the hypotheses that the cryosurvival of porcine embryos can be improved after partial delipation through chemically stimulated lipolysis and that the survival can be further improved by inhibition of apoptosis. Porcine embryos were produced in vitro using sow oocytes.