Hematologic assessment in pet rats, mice, hamsters, and gerbils: blood sample collection and blood cell identification.

Hamsters, gerbils, rats, and mice are presented to veterinary clinics and hospitals for prophylactic care and treatment of clinical signs of disease. Physical examination, history, and husbandry practice information can be supplemented greatly by assessment of hematologic parameters. As a resource for veterinarians and their technicians, this article describes the methods for collection of blood, identification of blood cells, and interpretation of the hemogram in mice, rats, gerbils, and hamsters.

Stress coping styles and singing behavior in the short-tailed singing mouse (Scotinomys teguina).

Stress coping styles have been characterized as a proactive/reactive dichotomy in laboratory and domesticated animals. In this study, we examined the prevalence of proactive/reactive stress coping styles in wild-caught short-tailed singing mice (Scotinomys teguina). We compared stress responses to spontaneous singing, a social and reproductive behavior that characterizes this species. To establish proactive/reactive profiles for singing mice, we measured exploratory and anxiety behavior using an open-field behavioral test. We examined correlations between open-field behaviors and fecal corticosterone (CORT) metabolites, baseline plasma CORT, and stress-induced CORT. Mice with proactive behavioral responses in the open-field had higher fecal CORT titers than reactive males, but did not differ in baseline or stress-induced plasma CORT. We suggest that individual differences in CORT metabolism may contribute to this surprising pattern. Males that sang in the open-field were behaviorally proactive and had lower stress-induced CORT, indicating a link between stress responses and singing in this species. Overall, the data demonstrate that singing mice offer an interesting model for exploring how stress reactivity can shape social behaviors.

Paternal behavior and testosterone plasma levels in the Volcano Mouse Neotomodon alstoni (Rodentia: Muridae).

Paternal behavior and testosterone plasma levels in the Volcano Mouse Neotomodon alstoni (Rodentia: Muridae). Although initially it was thought that testosterone inhibited the display of paternal behavior in males of rodents, it has been shown that in some species high testosterone levels are needed for exhibition of paternal care. In captivity, males of Volcano Mouse (Neotomodon alstoni) provide pups the same care provided by the mother, with the exception of suckling. Here we measured plasmatic testosterone concentrations 10 days after mating, five and 20 days postpartum, and 10 days after males were isolated from their families in order to determine possible changes in this hormone, associated to the presence and age of pups. Males of Volcano Mouse exhibited paternal behavior when their testosterone levels were relatively high. Although levels of this hormone did not change with the presence or pups age, males that invested more time in huddling showed higher testosterone levels. It is possible that in the Volcano Mouse testosterone modulates paternal behavior indirectly, as in the California mouse.

Paternal behavior and testosterone plasma levels in the Volcano Mouse Neotomodon alstoni (Rodentia: Muridae)

Paternal behavior and testosterone plasma levels in the Volcano Mouse Neotomodon alstoni (Rodentia: Muridae). Although initially it was thought that testosterone inhibited the display of paternal behavior in males of rodents, it has been shown that in some species high testosterone levels are needed for exhibition of paternal care. In captivity, males of volcano Mouse (Neotomodon alstoni) provide pups the same care provided by the mother, with the exception of suckling. Here we measured plasmatic testosterone concentrations 10 days after mating, five and 20 days postpartum, and 10 days after males were isolated from their families in order to determine possible changes in this hormone, associated to the presence and age of pups. Males of volcano Mouse exhibited paternal behavior when their testosterone levels were relatively high. Although levels of this hormone did not change with the presence or pups age, males that invested more time in huddling showed higher testosterone levels. It is possible that in the volcano Mouse testosterone modulates paternal behavior indirectly, as in the California mouse. Rev. Biol. Trop. 57 (1-2): 433-439. Epub 2009 June 30.

Aunque inicialmente se pensaba que la testosterona inhibía el despliegue de conducta paterna en los machos de roedores, en algunas especies se ha demostrado que son necesarios niveles altos de testosterona para la exhibición de cuidados paternos. En cautiverio, los machos del ratón mexicano de los volcanes (Neotomodon alstoni), proporcionan a sus crías los mismos cuidados que la madre, con la excepción del amamantamiento. En este estudio se midieron los niveles plasmáticos de testosterona: en el día 10 del apareamiento, 5 y 20 días postparto, y 10 días después de aislar a los machos de sus familias, para establecer si los niveles de esta hormona cambian con relación a la presencia y edad de las crías. El ratón de los volcanes exhibió cuidados paternos cuando sus niveles de testosterona fueron relativamente altos, aunque los niveles de esta hormona no variaron significativamente con relación a la edad y presencia de las crías. Sin embargo, los machos que invirtieron más tiempo en abrigar a las crías, presentaron concentraciones relativamente más altas de testosterona. Es posible que en el ratón de los volcanes la testosterona regule la conducta paterna de manera indirecta, como ocurre con el ratón de California.

Protein-energy malnutrition alters the C3 complement factor in response to lipopolysaccharide in a murine model / A desnutrição proteico-energética altera a concentração do fator C3 do sistema complemento em resposta ao lipopolissacarídeo em um modelo murínico

Malnutrition modifies the hostfis resistance to infection, altering many physiological processes, including hematopoiesis and immunological functions. In this study, we evaluated the total complement system and its C3 component factor in animals subjected to a protein malnutrition model with lipopolysaccharide (LPS) stimulation. The cellularity of blood, bone marrow and spleen, as well as the production of C3 and CH50, were evaluated. Two-month old male Swiss mice were submitted to protein malnutrition with a low-protein diet containing 4% protein. A control group received a control diet containing 20% protein. When the experimental group had reached a loss of about 20% in their original body mass, they were intravenously inoculated with LPS. Cells in the blood, bone marrow and spleen were counted and the circulating levels of C3 and CH50 were evaluated in these animals. Malnourished animals presented anemia, leucopenia, and a severe reduction in bone marrow and spleen cellularity. The survival rate of the malnourished animals was lower, as well as the production of C3 and CH50, if compared to the control animals. These findings suggest that malnourished mice present a deficient response to LPS. The decrease in the complement synthesis may be partially responsible for the immunodeficiency observed. These data lead us to conclude that the nutritional status interferes in the immune response activation.

La desnutrición proteico-energética modifica la resistencia a la infección, alterando numerosos procesos fisiológicos, incluyendo la hematopoyesis y la función inmunológica. En este estudio medimos las concentraciones séricas del factor C3 y del Sistema Complemento Total (CH50) en ratos con desnutrición proteico-energética estimulados con lipopolisacárido (LPS). Fue evaluada la celularidad periférica, medular y esplénica. Ratos Swiss, machos, adultos, con dos meses de edad fueron sometidos a desnutrición proteica con una dieta de 4% de proteína .El grupo control consumía una dieta con 20% de proteínas. Cuando los animales desnutridos perdieron 20% de su peso inicial, fueron inoculados por vía endovenosa con LPS. Fueron determinadas las concentraciones de células sanguíneas, de la médula ósea, del bazo y los valores de C3 y CH50 circulantes en los animales estimulados. Los animales desnutridos presentaron anemia y leucopenia además de una reducción significativa de celularidad de la médula ósea y del bazo. La sobrevivencia de este grupo era menor y también eran más bajas las concentraciones del factor C3 del complemento y del complemento total, en relación a los animales del grupo control. Los resultados sugieren que animales desnutridos muestran una respuesta deficiente a LPS. La síntesis menor de complemento puede ser en parte responsable por la inmunodeficiencia observada. Estos resultados nos conducen a inferir que la desnutrición proteico-energética interfiere en la activación de la respuesta inmune.

A desnutrição proteico-energética modifica a resistência à infecção, modi? cando diversos processos fisiológicos, incluindo a hematopoiese e as funções imunológicas. Neste estudo, avaliamos as concentrações séricas do fator C3 e do Sistema Complemento total (CH50) em um modelo no qual camundongos submetidos à desnutrição proteico-energética são estimulados com lipopolissacarídeo (LPS), e avaliamos a celularidade periférica, medular e esplênica. Camundongos Swiss, machos, adultos, com dois meses de idade foram submetidos ao processo de desnutrição proteica com uma dieta contendo 4% de proteína em comparação aos animais controles com uma dieta contendo 20% de proteína. Quando os animais do grupo desnutrido alcançaram aproximadamente 20% de perda de peso, em relação ao inicial, foram inoculados endovenosamente com LPS. As células do sangue, da medula óssea e do baço foram quantificadas, bem como as concentrações circulantes de C3 e CH50 em animais estimulados com LPS. Os animais desnutridos apresentaram anemia e leucopenia, além de redução significativa da celularidade da medula óssea e do baço. Os animais desnutridos apresentaram menor taxa de sobrevivência, bem como das concentrações do fator C3 do complemento e do complemento total em relação aos animais controles. Estes resultados sugerem que animais desnutridos apresentam uma resposta deficiente aos LPS. A síntese menor do complemento pode ser em parte responsável pela imunodeficiência observada. Estes resultados conduzem-nos a inferir que a desnutrição proteico-energética interfere na ativação da resposta imune.

Prolactin levels in paternal striped mouse (Rhabdomys pumilio) fathers.

Paternal behavior is associated with an increase in prolactin levels in fish, birds and mammals, including rodents. The striped mouse (Rhabdomys pumilio) from southern Africa shows highly developed paternal care. We investigated whether striped mouse fathers have higher prolactin levels than nonfathers, and whether there is a relationship between tactile stimulation with pups and prolactin secretion in fathers. We measured serum prolactin in 42 male striped mice assigned to one of four different experimental groups (single males, paired males, fathers housed with mother and pups, and fathers separated from their family by a wire-mesh partition). Our results revealed no increases in prolactin levels in fathers, and fathers with tactile contact with pups did not have higher prolactin levels than the fathers that were prevented from making tactile contact with pups. In contrast, experienced males had higher prolactin levels than inexperienced males. Male striped mice are polygynous in nature, living in groups, with three breeding females, and are permanently associated with pups during the breeding season. In a field study, males had higher prolactin levels during the breeding season than during the nonbreeding season. Thus, prolactin secretion in the polygynous striped mouse might be regulated by environmental stimuli, whereas social stimuli might be important for monogamous species. This is the first study to demonstrate seasonal changes in prolactin levels in a free-living male mammal.

Evaluation of an enzyme-linked immunosorbent assay for detection of antibodies to Junin virus in rodents.

Junin virus is the etiological agent of Argentine hemorrhagic fever, a serious rodent-borne disease. An enzyme-linked immunosorbent assay (ELISA) to detect Junin virus IgG antibodies in rodents was evaluated using sera from 27 Calomys musculinus and five Calomys laucha, inoculated experimentally with a live attenuated strain of this arenavirus. The test performance was compared against an indirect immunofluorescence assay (IFA). The ELISA had a sensitivity and specificity of 100% and a reproducibility of 87.9% for samples with titers above the selected cut-off value. IFA had lower sensitivity (53%) with the same specificity. The ELISA results were similar, whether carried out on whole blood or serum samples, thus eliminating the need for serum separation. A high correlation (K=0.86) between ELISA and IFA results was obtained from 1011 wild sigmodontine and murine rodents collected within and outside of the Argentine hemorrhagic fever endemic area. These results indicate that Junin virus IgG ELISA is the most suitable assay for detection of Junin virus antibodies in rodent samples.

Serum biochemical values in two inbred strains of mastomys (Praomys coucha).

Serum samples collected from 119 (72 male and 47 female) mastomys (Praomys coucha) of 2 specific-pathogen-free inbred strains (RI4 and RI7) were analyzed for 12 serum biochemical parameters. Sex-related differences (p < 0.01) were noted in alkaline phosphatase and glucose; the both higher in females than in males. Age-related changes (p < 0.01) were observed in total protein, albumin, total cholesterol, and alkaline phosphatase, with higher values for the first three parameters in the older group (200-250 days of age) than in the younger group (90-140 days of age). Four out of 12 parameters showed strain-related differences (p < 0.01), consistent with the large amount of genetic heterogeneity reported in this species. These serum biochemical reference values should provide information for the use of mastomys in laboratory research.

[DNA-protein cross-links in leukocytes of the field mouse (Apodemus agrarius) and the bank vole (Clethrionomys glareolus) dwelling in the territory of a radioactive waste storage station]. / DNK-belkovye sshivki v leikotsitakh polevykh myshei (Apodemus agrarius) i ryzhikh polevok (Clethrionomys giareolus), obitaiushchikh na territorii punkta zakhoroneniia radioaktivnykh otkhodov.

DNA-protein cross-links in white blood cells of Apodemus agrarius and Clethrionomys glareolus dwelling in the territory of the guarded zone of the radioactive waste storage in Sergievo Posad (area with gamma-background 50-400 microR/h) have been determined. As a control the animals from sanitary protected area of Sergievo Posadsky department (gamma-background on the soil surface is 8-10 microR/h) was used. It was found that the doses received by Apodemus agrarius and Clethrionomys glareolus, resulted in increasing of the DNA-protein cross-link level in the Apodemus agrarius white blood cells by a factor of 1.8, and in Clethrionomys glareolus by a factor of 1.4, in comparison with control. The increase of DNA-protein cross-link number reliably correlates with the beta-emitting nuclide content in animal bodies. The changes in the number white of cells and in the white blood composition were not observed.

Metal chelators/antioxidants: approaches to protect erythrocytic oxidative stress injury during Plasmodium berghei infection in Mastomys coucha.

Desferal, N-acetyl penicillamine (metal chelators) and propylgallate, catechin, and reduced glutathione (antioxidants) suppressed the erythrocytic oxidative damage generated during Plasmodium berghei infection in Mastomys coucha. Superoxide anion and lipid peroxide levels were increased and on the contrary, superoxide dismutase activity was noticeably decreased in the infected erythrocytes. Metal chelators/antioxidant treatment to infected animals resulted in restoration of O(2)(-), LPO and SOD to near normal levels. Furthermore, treatment of the above mentioned agents displayed a controlled mortality/survival time and parasitaemia. In conclusion, metal chelators/antioxidants were found to be effective against oxidative stress injury and parasite growth resulting in prolonged survival time of the host during experimental malaria.

A twenty strain survey and backcross localization of the erythrocytic GTP concentration determining locus Gtpc on mouse chromosome 9.

Erythrocyte nucleotide concentrations were surveyed among 20 inbred strains of mice in order to further assess the variability in GTP concentration. There was no significant difference in erythrocytic ATP concentration (Scheffé's test at P = 0.01), 678-1154 nmol/mL packed cells, among the strains surveyed. Two groups were distinguishable with respect to erythrocytic GTP concentration, 8 strains having high GTP, 215 +/- 44 nmole/mL packed cells, and 12 strains having low GTP, 34 +/- 12 nmole/mL packed cells. The erythrocytic GTP concentration determining trait Gtpc was previously shown to be linked to transferrin, Trf, on chromosome 9. Analysis of 232 [(B6 x WB) F1 x B6] backcross individuals for Gtpc and 8 microsatellite markers restricted the localization of Gtpc to a 5.6 +/- 2.1 cM region. The gene order and genetic distances in cM +/- SE are: (D9Mit14) 0.4 +/- 0.4 (D9Mit24) 1.7 +/- 0.8 (Gtpc, D9Mit51, D9Mit116, D9Mit212) 3.9 +/- 1.3 (D9Mit200) 3.0 +/- 1.1 (D9Mit20) 7.8 +/- 1.8 (D9Mit18). The GTP concentration determining trait appears to be a property of erythrocytes as no differences were observed for GTP/ATP ratios of brain, kidney, liver, and tongue from a low GTP strain, C3H/HeHa x Pgk-la and a high GTP strain, C57BL/6J.

Spontaneous and radiation-induced micronuclei in erythrocytes from four species of wild rodents: a comparison with CBA mice.

Almost 100 animals of 4 different species of small wild rodents (bank vole, Clethrionomys glareolus; field vole, Microtus agrestis; yellow-necked mouse, Apodemus flavicollis; and wood mouse, Apodemus sylvaticus) were trapped in central Sweden and used in experiments to determine the spontaneous and radiation-induced frequencies of polychromatic (fMPCE) and normochromatic erythrocytes (fMNCE) from bone marrow (bm) and peripheral blood (pb) using flow cytometric analysis. The results were compared with those from similar experiments with CBA mice. The saving of time and labour by the use of the flow cytometer-based analysis was a prerequisite for this study in which about 135 million PCE were analysed. The two species of voles had a mean background fMPCE (bm) of about the same value as CBA mice, while the yellow-necked mice had about five times higher fMPCE (bm). Wood mice had more than twice the fMPCE (bm) compared to CBA mice. Between individual animals in each of the 4 species, the background fMPCE (bm) varied more than between individual CBA mice, and the elimination of micronucleated erythrocytes was considerable. When exposed to ionizing radiation, the voles did not show a significant response. The response of the two Apodemus species was similar to that of the CBA mice, although it varied between individual animals and was not correlated to their background fMPCE. This study indicates that bank voles and field voles are unsuitable testing objects in the in vivo micronucleus assay. On the other hand, yellow-necked mice and wood mice seem to be useful in this test. Since the variation between individuals is considerable in wild Apodemus mice, large groups will be needed for obtaining statistically significant results when exposure to a genotoxic agent is low. Alternatively, repeated samples can be taken from individual wild mice to study the effect of a decreased exposure after keeping the animals for a period of time in an uncontaminated environment.

Effects of air pollution on hematological and plasma parameters in Apodemus sylvaticus and Mus musculus.

The effects of atmospheric air pollution on some hematological and plasma parameters, activity, and food and water consumption in Apodemus sylvaticus (wood mouse) and Mus musculus (mouse) were studied. The studies were undertaken in the area of Cercs (polluted area in Spain), where there is a coal-fired power plant that emanates SO2, NOx, and particles, and the area of St. Jaume de Frontanyà (nonpolluted area), located 40 km from the area of Cercs, with similar climatology, relief, and altitude, but without air pollution. Apodemus sylvaticus were captured in the field and Mus musculus were placed in cages in both studied areas. The results showed that there is a significant decrease in hematocrit, and a significant increase in leucocyte number, mean corpuscular hemoglobin concentration, osmolality, and gamma-globulin in Mus musculus, and a decrease in albumin (%) and albumin/globulin index in Apodemus sylvaticus. A negative correlation between hematocrit values and pollution was found. In mice placed in cages in the polluted area a slight but significant decrease in water and food consumption, and slower activity, was observed. The results suggest that pollution could induce a decrease in water and food consumption, which may be due to an observed reduction in activity.

[The genetic links and species affiliation of the field mouse (Rodentia, Muridae, Sylvaemus) from the Pamir-Alai]. / Geneticheskie sviazi i vidovaia prinadlezhnost' lesnoi myshi (Rodentia, Muridae, Sylvaemus) Pamiro-Alaia.

Analysis of variation in 30 presumptive biochemical loci in Sylvaemus sylvaticus pallipes from Tadjikistan was carried out with reference to the geographically closely related species S. uralensis. The data obtained support the genetic identity of pallipes, on the one hand, and suggest a rather low level of genetic divergence (D'Nei = 0.154, Pfd = 13) from an ancestral taxon, on the other hand. Analysis of genetic variation of diagnostic morphological features suggests that pallipes is the most extreme form of geographic population series of the uralensis species group. Both genetic and morphometric data suggest that S. pallipes is an allospecies of the uralensis species complex.

[A quantitative morphological study of the blood system in the common field mouse and the red-backed vole living in the area of the eastern Urals radioactive trace]. / Kolichestvenno-morfologicheskoe issledovanie sistemy krovi lesnoi myshi i krasnoi polevki, obitaiushchikh na territorii VURSa.

A number of quantitative morphological indices of blood system of two mice-like rodent species caught in radioactive contaminated areas was studied. Certain deviation in haemopoiesis and in blood characteristics were noted. The degree of deviations was not similar in the species with different radiosensitivity. It depends most probably on the total effect of the factors influenced animals in surroundings.

Litomosoides carinii: macrofilariae-derived glycolipids--chromatography, serology and potential in the evaluation of anthelminthic efficacy.

A preliminary characterization of the glycolipids of Litomosoides carinii macrofilariae, resolved according to their chromatographic, chemical and serological properties, has been performed. Emphasis has been placed on the neutral fraction glycolipids. These are separable on thinlayer chromatography into two groups of fast and slow migrating band components, that differ in their migration, differential chemical staining and serological traits, respectively. Serological analyses have been accomplished by thin-layer chromatography immunostaining and ELISA. Only components of the slow migrating band group react with infection serum from Litomosoides carinii-infected Mastomys coucha. Cross-reactivity experiments with homologous and heterologous infection sera of various helminthiases indicate that, epitopes bound to the neutral glycolipid fraction show structural similarity within the Nematoda, but not to the Cestoda or Trematoda. The dynamic development of specific Ig-, IgG- and IgM-anti-neutral glycolipid fraction antibody levels were correlated with the different progression of L. carinii and Brugia malayi infections in the multimammate rat, Mastomys coucha. The reduction in the dynamics of IgG- and IgM-antibody levels on chemotherapeutic treatment with the filaricides flubendazole and CGP 20376 has been related to their macrofilaricide-activity.

[Number and maturation distribution of the reticulocytes from six animal species]. / Zahl und Reifegradverteilung der Retikulozyten von sechs Tierarten.

With the fully automated reticulocyte analyzer Sysmex R-1000 (Fa. Toa Medical Electronics Co., LTD) the reticulocyte count and the reticulocyte ratio of rabbit, rat, mouse, mastomys, dog and swine was determined. Simultaneously, the reticulocytes were subdivided into 10 maturation classes according to their fluorescence intensity. The maturation distribution can be characterized by a distribution quotient. Both the maturation distribution as well as the distribution quotient are considered to be useful as good indicators of erythropoietic activity. The investigation establishes, that the reticulocyte count and the maturation distribution of these six species can be determined quickly and easily. The distinct differences between the species reflect the maturation locus of the reticulocytes, which varies among the species (medullary or humoral type of maturation).

Characterization of third beta-chain variant in Rattus rattus rufescens.

Different molecular forms of hemoglobins of locally available murines, represented by Rattus rattus rufescens, have been investigated and the probable genetic mechanisms leading to the observed heterogenicity in the hemoglobin phenotypes are discussed. Each fraction was isolated in chromatographically pure form, identified and characterised structurally to establish their alpha- and non-alpha-chain constitution. Six molecular forms of component hemoglobins were identified from a wild population of R. rattus rufescens. The present study suggests five different globin chains in the hemoglobin of house rats (Rattus rattus rufescens). There are apparently two alpha-chains, namely alpha I and alpha II, and three different beta-chains, viz. beta I, beta II and beta III. The invariable presence, though at varying concentrations, of all these five globin chains implicates a gene duplication at the alpha-chain loci and a gene triplication at the beta-chain loci, the latter being a rather rare and unique genetic event.

Mastomys natalensis or Mastomys coucha. Correct species designation in animal experiments.

Haemoglobin electrophoresis and chromosome analysis indicate that the widely used multimammate rat (chamois coloured) as experimental animal belongs to the species Mastomys coucha (Smith, 1836) and not, as commonly assumed, to Mastomys natalensis (Smith, 1834).