High-Containment Agriculture Animal Research: An AAALAC International Perspective.

Institutions that conduct high-containment agricultural research involving domestic livestock represent a specialized category of programs that are accredited by AAALAC International. The accreditation process includes a comprehensive assessment of the overall program of animal care and use. However, the complex design of these facilities and the unique care required for animals in this type of environment often mean that additional attention will be directed at areas regarded as higher risk when the programs are evaluated. Specific issues that may stimulate additional discussion and interest include animal housing practices, environmental conditions inside the facility, maintenance of procedure and support areas, methods for obtaining and safely transporting healthy research animals, strategies to minimize animal pain and distress, unusual protocol review challenges, and institutional policies relevant to personnel training and safety. These issues are further discussed to inform institutions of potential concerns that should be reviewed and assessed during internal preparations for accreditation visits by AAALAC site visit teams.

Disaster Medicine: Disaster planning for agricultural research programs.

Institutions that conduct agricultural research must plan for emergencies and disasters that have the potential to compromise the health and safety of research animals and personnel. Agricultural research facilities have unique challenges owing to the overall size and scope of operations, wide range of species housed, and various types of facilities maintained. Identification of hazards and development of strategies to minimize anticipated risks are important to creating a successful mitigation and recovery plan that will minimize both short- and long-term adverse effects on program operations and resources.

Flow cytometric analysis of micronuclei in peripheral blood reticulocytes III. An efficient method of monitoring chromosomal damage in the beagle dog.

Erythrocyte-based micronucleus tests have traditionally analyzed bone marrow because splenic filtration in most species removes micronucleated cells from peripheral blood. We have evaluated a flow cytometric method for monitoring micronucleated reticulocyte frequencies (%MN-RET) in the peripheral blood of beagle dogs treated with cyclophosphamide (CP) and have found that analysis of micronucleated reticulocytes (MN-RETs) in peripheral blood is a suitable surrogate for bone marrow analysis. The three-color flow cytometric method uses anti-CD71 labeling to identify reticulocytes and Plasmodium berghei-containing erythrocytes as a calibration standard. The spontaneous %MN-RET determined by flow cytometry was 0.31 +/- 0.09% (n = 22) for peripheral blood, compared with 0.38 +/- 0.13% (SD, n = 12) for bone marrow, and 0.27 +/- 0.08% (n = 12) for peripheral blood by microscopic scoring with acridine orange staining. The kinetics of appearance and disappearance of MN-RETs in blood were determined by collecting daily samples after iv treatment with CP. The maximum frequency occurred approximately 48 h after dosing. Frequencies of MN-RETs in peripheral blood at steady state following daily CP treatment were 55-68% of corresponding bone marrow values assessed by microscopy and 55-112% as assessed by flow cytometry. This difference is presumably due to splenic removal, which appears slightly less stringent than that previously reported for CP-treated Sprague-Dawley rats. Responses in bone marrow and peripheral blood were highly correlated and similar to or greater than those reported in mice and rats at equitoxic doses.

Characterisation of macrophage migration inhibitory factor from Eimeria species infectious to chickens.

Macrophage migration inhibitory factor (MIF) was the first cytokine to be identified almost 40 years ago. Homologues of MIF have been isolated recently from invertebrates, making it an interesting molecule from an evolutionary as well as functional perspective. The present study represents the first report of MIF homologues in apicomplexan parasites, belonging to the genus Eimeria. A single full-length clone was isolated from Eimeria acervulina that shared between 35 and 38% amino acid identity with MIFs of vertebrates. A MIF cDNA from Eimeria tenella shared 64% amino acid identity with E. acervulina MIF. The mRNA expression was highest in merozoites, whereas developing oocysts and sporozoites expressed low to undetectable levels. Protein expression patterns were nearly identical to that observed by reverse transcriptase polymerase chain reaction (RT-PCR), suggesting strong developmental regulation. Immunofluorescence staining and co-localisation studies of E. acervulina merozoites indicated that MIF is distributed throughout the cytosol, and appears to be concentrated in the apical end of the parasite. The presence of MIF was detected in excretory/secretory (ES) products collected from E. acervulina merozoites, and isoelectric focusing indicated that three MIF isoforms are present in this stage. Phylogenetic analysis revealed that apicomplexan MIF sequences form a sister relationship to MIF-like molecules from Arabidopsis thaliana.