Husbandry and medicine of small rodents

Biology, physiology, husbandry, and commonly occurring diseases of small rodent pets, including the Syrian hamster, Mongolian gerbil, rat, and mouse, are presented.     

Different patterns in the histology and autofluorescence of the Harderian glands of the Syrian Hamster, rat, mouse, Mongolian gerbil and guinea pig

The purpose of this study was to compare the natural fluorescence in the Harderian glands of the Syrian hamster, rat, mouse, Mongolian gerbil and guinea pig (both sexes). For each species, 10 animals (five males and five females) were used. Histological autofluorescence studies were performed using a fluorescence microscope (450-490 nm filter). Two different types of fluorescent cells were observed in both hamster (type AFI high intensity and type AFII, low fluorescence) and rat (type AFI, low fluorescence and type AFII, high fluorescence) Harderian glands. The fluorescence was basally located in all mice cells, whereas it was observed near the epithelial cell nuclei in the Mongolian gerbil (occupying two-thirds and one-third of the cells in males and females, respectively). A high intensity of fluorescence was present throughout the acinar cells in the guinea pig. The patterns of fluorescence identified exhibited a sexual dimorphism in all species studied. These results demonstrate that the Harderian glands of the animal species examined exhibit a variety of histological autofluorescence patterns.     

In vitro progesterone production of corpus luteum in pregnant Mongolian gerbils (Meriones unguiculatus)

The present study was conducted to determine the in vitro progesterone (P₄) production by Mongolian gerbil (Meriones unguiculatus) corpus luteum (CL) on days 9, 18 and 22 of pregnancy in comparison with rat CL on days 9 and 16. The culture medium of one CL per well of a 96‐well plate was changed after 4 h incubation and the CL were further cultured for 24 h. The P₄ in serum and in the medium were measured by enzyme immunoassay system. Gerbil CL weight measured after 24 h incubation was at the same level for all 3 days of pregnancy and significantly different from that of the rat CL for the 2 days of pregnancy. The serum P₄ level in gerbils was high in early pregnancy and decreased in advanced days of pregnancy. In contrast, the P₄ in rats was higher in the latter half of pregnancy. In vitro P₄ production by a single CL after 4 h incubation decreased significantly during the days of pregnancy in both the gerbil and the rat. P₄ values produced by the gerbil CL for 24 h incubation were significantly higher than after 4 h incubation in all 3 days of pregnancy, however, there was no difference between the two incubation time groups in the rat. In vitro P₄ production by CL correlated well with the serum P₄ level on the 3 days of pregnancy in the gerbil. In the rat, however, the relationship was inversed between the two parameters. The correlation in this experiment suggested that CL is the sole source of P₄ for the maintenance of the gerbil pregnancy, with the result of abortion after an ovariectomy on day 20 of pregnancy as described in our previous report.     

Effects of levonorgestrel‐quinestrol (EP‐1) treatment on Mongolian gerbil wild populations: a case study

Rodent pest population outbreaks occur frequently in grassland ecosystems in northern China. The Mongolian gerbil (Meriones unguiculatus) is a dominant pest rodent which is distributed across the semi‐desert grasslands of Inner Mongolia, China. In 2009, we studied the contraceptive effect of levonorgestrel‐quinestrol (EP‐1), concentration 50 ppm, on a wild Mongolian gerbil population. The one‐off contraceptive treatment was compared with a control group using a semi‐monthly live trapping method in the Ordos Semi‐desert Grassland Region of Inner Mongolia. The results show that juveniles were not recruited in spring in the treatment group. Ratios of juveniles in the control and treatment groups showed significant semi‐monthly differences from spring to summer (one‐way ANOVA, F_{2, 14}= 7.53, P < 0.05). Between both groups, annual fluctuations of juvenile and total population densities were significantly different respectively (F_{2, 14}= 4.64, P < 0.05; F_{2, 18}= 7.72, P < 0.05). The contraceptive EP‐1 delayed the normal reproductive pattern of Mongolian gerbil populations. This suppressed birth rates of gerbil populations, reduced their densities, and changed their age structures. The period of EP‐1 baiting should be extended but it could be an ideal method for controlling Mongolian gerbil populations during each breeding season.     

Characterization of monoclonal antibodies directed against swine leukocytes

Hybridomas were produced from fusions of the SP2/0 mouse myeloma with splenic cells from: 1) an outbred Sprague Dawley rat immunized with swine peripheral blood mononuclear (PBM) cells; 2) a (CBA/NDub X BALB/c Dub) F1 mouse immunized with concanavalin A (Con A) activated swine PBM cells and 3) a (BALB/c Dub X C3H/He Dub) F1 mouse immunized with swine thymocytes. The resulting supernatants were screened by a microcytotoxicity assay for activity against swine PBM cells. Four hybridomas (MSA1, MSA2, MSA3 and MSA4) were selected, cloned and characterized by their cell reactivity and effect on mitogenic assays. MSA1 and MSA2 belong to the rat IgG2b subclass. MSA3 and MSA4 are of the mouse IgG2a subclass. These monoclonal antibodies reacted in the following manner: MSA1 with monocytes, granulocytes, red blood cells and bone marrow cells; MSA2 with subset of T cells; MSA3 with B cells and subsets of T cells and monocytes (class II molecule) and MSA4, a pan-T cell reagent (E-rosette receptor). The involvement of the various cell types reactive to the different monoclonal antibodies in the mitogenic response of swine PBM cells to Con A, phytohemagglutinin (PHA) or pokeweed mitogen (PWM) was investigated by cellular depletion with monoclonal antibody plus complement. Cellular depletion of PBM cells with the following monoclonal antibodies plus complement treatment resulted in: MSA1, almost total reduction in the mitogenic response to low doses of Con A or PWM; MSA2, partial reduction in the proliferative responses to any concentration of Con A, PHA or PWM; MSA3, partial reduction in proliferative responses to low concentrations of Con A or PWM and 4) MSA4, total elimination of any proliferative response to Con A, PHA or PWM.     

Vergleich der histologischen Struktur der Kompakta der langen Röhren-knochen bei Maus, Hamster, Ratte, Meerschweinchen, Kaninchen, Katze und Hund während der Altersentwicklung

This paper reports on histological examinations of the diaphyseal structure of the long bones (humerus, radius, ulna, os femoris and tibia) of mouse, hamster, rat, guinea pig, rabbit, cat, and dog using six animals in three different age groups. There are considerable differences in structure between the species. The results show that with increasing height in the evolution of the species and period of life, as well as advancing age, the differentiation and complexity of bone structures increases. The differences in structure between the localization of bones within the individual species are not considerable. The species comparison results in great similarities between the bone structures of each of mouse and hamster, of rat and guinea pig, as well as of cat and dog. The bone structure of the animals examined becomes more similar to the human structure from mouse, hamster, rat, guinea pig, rabbit, and cat to dog.     

Cryopreservation and In Vitro culture of Preimplantation Embryos in Djungarian Hamster (Phodopus sungorus)

Although embryo cryobanking was applied to Syrian golden and to Campbell's hamsters, no attempt has been made at freezing embryos in Djungarian hamsters. Four‐cell stage embryos were flushed from the reproductive ducts of pregnant females before noon of the third‐day post coitum and frozen in 0.25‐ml straws according to standard procedures of slow cooling. A mixture of permeating (ethylene glycol) and non‐permeating (sucrose) cryoprotectants was used. The thawing was performed by incubating at RT for 40 s followed by 40 s in a water bath at 30.0°C. Most (66.7%) of the non‐frozen four‐cell embryos developed up to the morula stage in rat one‐cell embryo culture medium (R1ECM). The use of hamster embryo culture medium (HECM) yielded fewer morulas (18.2%) during the same 24‐h period of culture. The rate of embryo's surviving the freezing–thawing procedures, as estimated by light microscopy, was 60.7–68.8%. After 24‐h culturing in R1ECM, 64.7% of frozen–thawed four‐cell embryos developed and all of them reached the morula stage. Supplementation of R1ECM with GM‐CSF (2 ng/ml) improved the rate of Djungarian hamster frozen–thawed embryo development: 100% of the four‐cell stage embryos developed, 50% of them achieved the morula stage, and 50% developed even further and reached the blastocyst stage within 24 h of culturing. This study reports the world's first successful transfer of frozen–thawed Djungarian hamster embryos yielding term pups. Taken together, the results of this study demonstrate the possibility of applying some key reproductive technologies, that is, embryo freezing/cryopreservation and in vitro culture, to Djungarian hamsters.     

Antibody responses in pregnancy-induced transmammary transmission of Ancylostoma caninum hookworm larvae

Third stage larvae of the Ancylostoma caninum hookworm nematode have the capacity to infect a dog, abort the normal maturation pathway to become blood-feeding intestinal worms, and instead distribute throughout the body in a developmentally arrested state that is relatively resilient to most chemotherapeutic agents. During pregnancy, a percentage of the arrested larvae reactivate and transmit via the mammary glands to infect the nursing puppies with resulting iron-deficiency anemia and potential mortality. To determine if the suppression of parasite-specific antibody responses during pregnancy facilitates the reactivation and transmammary transfer of hookworm larvae, a murine model of A. caninum infection was used to compare the infected versus uninfected animals that were either bred or not bred. Initial comparisons of genetically divergent BALB/c versus C57BL/6 mice showed that both the strains mounted strong Th2 biased IgG1and IgE antibody responses to A. caninum infection. Using the BALB/c strain for the breeding analyses, it was confirmed that larval transfer to the mouse pups only occurred during the post-partum lactational period. In the dams, levels of total and antigen-specific IgG1 and total IgE were highly correlated with parasite burden. During most phases of pregnancy and lactation, infected dams had lower total IgG1, IgG2a and IgE levels as compared to unbred mice at comparable times post-infection; this downward modulation of antibody responses supports the established dogma of a generalized immunosuppression associated with pregnancy. However, at parturition and post-partum lactation, antigen-specific IgG1 levels measured at 1 : 5000 serum dilutions were comparable between bred and unbred mice, and antigen-specific IgG2a levels at 1 : 100 serum dilutions were also not significantly different except for a marginal reduction in the bred mice at the lactational timepoint. The comparable anti-A. caninum IgG1 levels between bred and unbred mice, and low correlation between IgG2a levels and larval burden suggest that parasite-specific antibody responses do not play a major role in the pregnancy-associated transmammary transmission of A. caninum larvae. This conclusion does not rule out the possibility that underlying fluxes in the levels of specific cytokines associated with pregnancy and infection may be involved in the process of larval reactivation and transmission.     

Characterization of a feline homologue of the alphaE integrin subunit (CD103) reveals high specificity for intra-epithelial lymphocytes

The characteristics of a feline homologue of the alphaE integrin (CD103), defined by two murine monoclonal antibodies, Fe7.1B8 (IgG1) and Fe7.2D8 (IgG1), are described. These antibodies recognized 75% of intra-epithelial (range 59-88%) and 40% of lamina proprial (range 28-46%) T cells of the intestinal mucosal tissue of the small intestine in contrast with approximately 2% of peripheral blood lymphocytes. Both antibodies immunoprecipitated a 180 kDa protein from biotinylated feline intra-epithelial mucosal leukocytes consistent with the alphaE integrin subunit in conjunction with a 120 kDa protein consistent with the beta7 subunit. The nucleotide sequence of feline alphaE integrin, generated from molecular cloning of the feline alphaE encoding cDNA, is also reported. This feline molecule shares 72% sequence homology with human and 69% homology with murine and rat counterparts. Homology includes the presence of an X (extra) domain, that appears unique to alphaE molecules as described for human, rat and mouse, as well as areas of homology common to other alpha integrins. Of note is a typical I (inserted) domain, the presence of seven repeat regions, and highly conserved sequences in the cytoplasmic tail. Transfection studies demonstrated that both antibodies recognized an extracellular component which encompassed the X and I domains of the cloned alphaE integrin subunit. These studies demonstrate that the pattern of tissue distribution, biochemical characteristics, and cDNA sequence of the feline alphaE integrin subunit are largely similar to that described for other species.     

Lactate dehydrogenase and creatine phosphokinase isoenzymes in tissues of laboratory animals

The lactate dehydrogenase (LDH) and creatine phosphokinase (CPK) isoenzyme distributions in tissues of the ICR mouse, Wistar rat, guinea pig and golden hamster were analyzed by histoelectrophoresis. Tissues obtained were as follows: liver, pancreas, stomach, small intestine, heart, femoral muscle, uterus, kidney, spleen, lymph node, cerebrum, spinal cord and erythrocyte. Histoelectrophoresis was for the direct analysis of LDH and CPK isoenzymes in the tissues and had high practical value compared with previous tissue-extraction methods. In tissues of the mouse, guinea pig and golden hamster, LDH isoenzymes showed five bands. In the rat, LDH isoenzyme was separated into four fractions. CPK isoenzyme showed three bands; BB, MB and MM. In some tissues, the MM band was separated into two sub fractions.     

河西走廊北部荒漠草地长爪沙鼠防治指标研究

长爪沙鼠是河西走廊北部荒漠草地的优势鼠类,危害严重。古浪县位于河西走廊东部,北部荒漠草地的长爪沙鼠,5月鼠密度在29．1只／hm^2～52．3只／hm^2之间,至9月造成牧草损失586kg／hm^2。通过调查鼠密度和牧草损失量,结果充分显示长爪沙鼠的种群密度与其造成的损失量之间呈明显的正相关关系,其回归方程式为：M=13．96×-8．10。通过防治及综合分析研究后提出在河西走廊的古浪县北部荒漠草地长爪沙鼠的防治指标为5．33只／hm^2,5月份防治。     

Comparative study on calretinin immunoreactivity in gerbil and rat retina

Expression of calretinin in retina has been ascribed to multiple biological and functional aspects in the visual system. In this study, we examined the distribution patterns of calretinin immunoreactivity in gerbil and rat retina. In the gerbil, calretinin immunoreactivity was present in bipolar and amacrine cells of the inner nuclear layer and in neurones of the ganglion cell layer. In the rat, amacrine and ganglion cells showed calretinin immunoreactivity, but bipolar cells did not contain calretinin immunoreactivity. In both species, calretinin immunoreactivity was absent in cones, cone bipolars, and horizontal cells. In conclusion, gerbil as well as rat has a rod-dominant retina. The differences in calretinin expression between rat and gerbil require further investigations under various functional and developmental conditions.     

Applicability of vital staining and tissue clearing to vascular anatomy and melanocytes' evaluation of temporal bone in six laboratory species

The purpose of the present study was to define the applicability of tissue clearing to the field of otology. We combined tissue clearing with vital staining perfusion via a pumping system to examine the vascular anatomy of temporal bones in laboratory animals. We used six different types of species including Korean wild mouse, mouse, Mongolian gerbil, hamsters and Guinea pigs. A mixture of Alcian blue reagent and 4% paraformaldehyde was circulated throughout the entire circulatory system of the animal via a perfusion pump system. Transparency images were obtained from the temporal bones according to the protocol of the SunHyun 3D Imaging Kit. In examining the inner surface of the tympanic membrane, flaccid part (pars flaccida) was positioned along the entire marginal area in Guinea pig. In the Guinea pig, unlike the other species, the cortical bone of the mastoid (bullae) was easily removed using cold instruments, allowing a direct approach to the enclosed structures. The distribution and pattern of cochlea melanocytes were compared among the species. “Mobius strip”‐like accumulated melanocytes in vestibules were shown in both the Korean wild mouse and mouse. The collateral blood supply to the cochlea in six different species was checked in various pattern. Combining dye infusion with tissue‐clearing techniques, we documented the middle ear and transparent inner ear structures in six different species. The information and associated images will help other researchers to develop hypotheses and design experimental investigations.     

Comparative studies on the distribution of glutamate transporters in the retinae of the Mongolian gerbil and the rat

Glutamate is the major excitatory amino acid transmitter in vertebrate retinae. Glutamate transporters therefore play an important role in the precise control of glutamate concentration in the synaptic cleft by regulating extracellular glutamate concentration. In the present study, we performed an analysis of the expressions of three glutamate transporters in gerbil retina using immunohistochemistry. In the gerbil retina, excitatory amino acid carrier 1 and glutamate transporter 1 immunoreactivity was predominant in the ganglion cells but not amacrine or bipolar cells. Glutamate/aspartate transporter (GLAST) immunoreactivity was observed in the radial gliocytes of which the dense network of fine processes was localized in the inner and outer plexiform layers. GLAST immunoreactivity was also detected in astrocytes in the nerve fibre layer. These results demonstrate that three glutamate transporters show specific distributions in the gerbil retina and suggest that the glutamate re-uptake system in the gerbil retina may be different from that of the rat.     

Comparative studies in the promoter and exon 1 regions of tumour suppressor p53 in several mammalian species: absence of mutations in a panel of spontaneous domestic animal tumours

Tumour suppressor p53 is critical in a broad panel of tumour types in human, mouse and other mammals. Regions of the promoter and exon 1 play an important role in expression of p53. In the present study, the DNA sequences of promoter and exon 1 regions of four domestic animal species (dog, cat, horse and cattle) are determined and compared with experimental rodents (mouse, rat and hamster) and man. A broad panel of tumour types have been investigated for mutations in this regulatory area in 90 canine, 136 feline, 25 equine and 10 bovine patients. No mutation was detected in any of the tumours analysed.     

A comparative study on cardiac ganglia in midday gerbil,Egyptian spiny mouse,chinchilla laniger and pigeon

Using the thiocholine method and histological techniques, the topography and morphology of cardiac ganglia in midday gerbil, Egyptian spiny mouse, chinchilla laniger and pigeon were studied. The results demonstrated that cardiac ganglia in all investigated species are embedded in epicardial fat. They formed plexo-ganglionic structures. Each of them composed of many ganglia (from seven up to 36) different in size and shape, and interconnected by fascicles of nerve fibres. Comparative analysis showed that the density of neural network and cell aggregations was different in individual species. The richest plexo-ganglionic structure was in pigeon. It was organized in three plexo-ganglia with an average of 30 ganglia. The largest one was located along the anterior interventricular sulcus. The cardiac ganglia of investigated mammals were localized mainly on the epicardium of atria; in Egyptian spiny mouse and chinchilla laniger on the ventral surface of right atrium, but in midday gerbil on the dorsal surface of left atrium. Moreover, in midday gerbil and Egyptian spiny mouse the little plexo-ganglionic structure on the ventricle were noticed. Additionally, in midday gerbil the single nerve cells might be observed between cardiac muscle of atria. It can be said that, the strongly developed cardiac plexus in pigeon is probably connected with his behaviour and functional properties of the heart. The arrangement of neurones in cardiac ganglia of all examined mammals was uniform over the whole surface of the sections, while in the pigeon, neurones were located mainly in the peripheral part of the ganglion.     

Generation of anti‐porcine CD69 monoclonal antibodies and their usefulness to evaluate early activation of cellular immunity by flow cytometric analysis

T cell‐mediated cellular immunity and humoral immunity are equally important for the prevention of diseases. To assess activation of human and mouse cellular immunity, early activation markers of lymphocytes are often used in flow cytometry targeting expression of CD69 molecules. Response of humoral immunity against infection or vaccination has been well investigated in pigs, but that of cellular immunity has been largely neglected due to lack of direct evaluation tools. Thus, in pig research a proper assay of antibody reacted with porcine CD69 is still unavailable. In the present study, two anti‐porcine CD69 mAb‐producing mouse hybridomas, 01‐14‐22‐51 (IgG2b–κ) and 01‐22‐44‐102 (IgG2a–κ), both showing fine reactivity with phorbol 12‐myristate 13‐acetate (PMA) and ionomycin‐stimulated porcine peripheral blood lymphocytes in flow cytometry, were established. When porcine peripheral blood lymphocytes were activated with PMA and ionomycin and analyzed by flow cytometry, it was found that both mAbs generated in this study stained about 70% of lymphocytes. In contrast, after an identical procedure, only 5% and 13.5% of lymphocytes were stained with anti‐interferon‐γ mAb and anti‐tumor necrosis factor‐α mAb, respectively. These results indicate that evaluation of cellular immunity activation turns more sensitive after using our newly generated mAbs.     

A recombinant-based ELISA evaluating the efficacy of netobimin and albendazole in ruminants with naturally acquired fascioliasis

The therapeutic efficacy of albendazole and netobimin in ruminants with naturally occurring fascioliasis was investigated using a recombinant-based ELISA. The variation in the IgG response against a 2.9-kDa recombinant protein (FhrAPS), termed efficacy index (EI) 1, and the egg-output changes, termed EI 2, were used to evaluate drug efficacy.The values of EI 1 ranged between 0% and 50% in sheep, and between 0% and 30% in cattle after treatment with albendazole and netobimin. Similar EI 2 values were observed in sheep receiving albendazole or netobimin, but the highest values were found in cattle treated with netobimin. The significant reduction in the IgG response to FhrAPS found in this study shows promise in terms of developing alternative methods for evaluating the efficacy of chemotherapy against Fasciola hepatica in grazing ruminants.