Abstract

Microbiological control of laboratory animals is important in any animal facility for production of microbiologically clean animals. It is possible through accurate diagnosis of the rodent pathogens in the laboratory. Pseudomonas aeruginosa is a ubiquitous and free living organism which infects the gastrointestinal tract through water. Normal animals do not show clinical signs in infection but immuno-compromised/ irradiated mice show varied symptoms of the disease. In this study, Pseudomonas aeruginosa was isolated from drinking water provided to the laboratory animals. The P. aeruginosa grown on the Nutrient and MacConkey agar plates were subjected for identification by using colony characters, odor and biochemical tests. DNA was isolated and PCR was carried out by using positive DNA as positive control. PCR product of 726 bp pairs was developed on 2% agarose gel for confirmation of the organisms. PCR method along with conventional method may offer sensitive and rapid detection of P. aeruginosa in water as well as clinical specimens.

Abstract

The objective of this study was to compare changes in leucocyte and lymphocyte analytes in various models of immunodeficient mice lacking T or B or NK cells or both T and B cells. In this study, we used the following immunodeficient mice (nu; T inactive B+ NK+), (IgH-6^-/-; T+B inactive NK+) (beige; T+B+NK inactive) and SCID and RAG-1^-/- (T inactive B inactive NK+). Among the T cell deficient (Ii^-/-, CD8+, CD4 Inactive) and (TAP-1^-/-; CD 4 inactive and CD8⁺) were used. FACS analyses of peripheral-blood mononuclear cells were performed to determine the percentage of CD3⁺ T cell, B220⁺ B cell and NK cell along with analysis of hematological parameters. There were marked differences in the relative proportions of leucocytes and lymphocytes blood cell population among the immunodeficient strains. These results indicate that WBC and lymphocytes population in whole blood depends on T cells percentage. B cells and NK cells deficiency has minor role in the leucocytes and lymphocytes population in immunodeficient status in mouse models. The hematological differences described here are based on the level of CD3, B220 and NK1.1 cells. This study will provide baseline information for researchers who use various immunodeficient mice for immunological, genetic and cancer studies.

Abstract

Immunophenotyping by flow cytometry is one of the most rapid way of doing analysis & identification of heterogeneous populations of cells by using cell-specific fluorochrome-conjugated antibodies as probes. Immunophenotyping of laboratory animals is essential to monitor the immune status of the laboratory animals for better maintenance and management of animal facilities for immunological studies. The spleen is one of the major organs in immunity and plays a key role in the production and maintenance of red blood cells and the production of certain circulating white blood cells. Therefore, we harvested the spleen from various mice strain (Swiss Albino, BALB/C, C57BL, and C3HeJ) available at Central Animal Facility of Indian Institute of Science and did quantification of various types of immune cells by multicolor flow cytometry. 1X10⁷ splenocytes were taken in the eppendorf tubes and labeled with T cell, B cell, CD4⁺ and CD8⁺ cells specific antibodies and 30,000 events were acquired in the BD FACS Canto™ II (BD Biosciences) and the results were analyzed by FACS Diva software version 6.1.1. It was found that C57BL has higher percentage of B Cells (60±10%) followed by C3HeJ (49±2%). BALB/C has higher number of T cells (54±4%) followed by Swiss Albino strain (46±4%). BALB/C has higher percentage of CD4⁺ Cells (39±3%) as compared to other strains. In most of the strains the B Cells population is more followed by T Cells, CD4⁺ Cells & CD8⁺ Cells. Moreover, splenocytes percent profile of Scid and Nude mice were taken as standard.

Abstract

Small laboratory animal models are being increasingly used as a biomedical research tool in several human diseases such as cancer, cardiovascular diseases, endocrine disorders and obesity. For the last couple of years, molecular and functional imaging become a powerful techniques for studying spatial and temporal distribution of new drugs and their target affinity non-invasively in animal models. Hence non invasive imaging modalities such as positron emission tomography (PET), single photon emission computed tomography (SPECT), X-ray, magnetic resonance imaging, fluorescence and bioluminescence have emerged for rapid and accurate screening of the new molecules. This review focuses on nuclear imaging modalities mainly SPECT and PET, which allow whole body survey of the animal and quantitation of target affinity. While establishing such facilities, accessibility to vivarium, work flow, availability of radiotracers, regulatory requirements and trained human resource needs to be considered. The readout data requires dedicated software for viewing and quantitation eg. VIVID, PMOD, AMIDE and Micro View. Therefore, a perfect amalgamation of instrumentation, facility logistics and work flow that is required for efficiently managing such a facility has been elaborately reviewed in this article.

Abstract

Laboratory zebrafish (Danio rerio) has proved promising vertebrate animal model in modern biology research during past few decades. Several inherent biological features in zebrafish, have invited attention of the scientific community to use them in genetics and developmental biology research. Attempts were made to establish facility to raise and maintain several lines of wild type and mutant zebrafish at the Department of Biological Sciences in 2009. A small room in the department housed ~50 wildtype zebrafish obtained from the ornamental fish shop at the local market (M/s. Vikrant Aquaculture, Mumbai). At the beginning, fish were raised in glass aquarium of 50 liters capacity for 6 to 12 months. During the period of initial setup, all the members of the group associated with the usage of zebrafish,received training in the zebrafish husbandry, water quality control, fish breeding and health monitoring. The state of the art facility was established later on by setting Recirculating Water System (RWS) procured from the Aquatic Habitat, USA. The wild type (WT) and mutant fish lines procured from the Max Plank Institute (MPI), Germany,were kept in RWS after ensuring healthy aquatic environment for the fish. The standard operating protocols for fish husbandry, breeding, nutrition, water and health quality of zebrafish were formulated.The breeding parameters such as fecundity, egg spawning, viability and mortality were studied. The female fecundity percentage noticed between 42 to 58 and the mean egg spawn found between 139.77 to 242.42 eggs per female in five strains. In-vitro culture of embryos revealed maximum mortality on first day of culture with decreased mortality on subsequent days. Mortality percentage observed during the period of culture ranged between 14.35 to 30.21 in five strains with mean mortality of 21.34%. The embryo viability in five strains ranged between 69.78% to 85.64% with mean viability of 78.65%. In the present study, attempts have been made to generate basic data associated with breeding of the laboratory zebrafish

Abstract

India is one of the countries which have oldest regulations for animal welfare and experimentation. The Committee forthe Purpose of Control and Supervision of Experiments on Animals (CPCSEA) was constituted by the Government ofIndia, way back in 1968 under chapter IV of the Prevention of Cruelty to Animals (PCA) Act, 1960 to oversee the animalexperiments and welfare.

Abstract

Corncob has been the most common choice of bedding material for laboratory animals in many countries due to its ready availability, low cost and better absorbance properties. However, corncob has been reported to have estrogenic, endocrine disruption and mitogenic properties by few authors. Animal facilities need to make a wise decision while selecting a particular bedding type, such that the unwanted effects associated with bedding like corncob, may not interfere with the research objectives. Chip bedding made from hardwood is one of the most common choices of bedding by most international animal facilities for commercial breeders, academic research institutes, pharmaceutical industry and preclinical contract research organisations.