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Borrelia burgdorferi ospC heterogeneity among human and murine isolates from a defined region of northern Maryland and southern Pennsylvania: lack of correlation with invasive and noninvasive genotypes.

B. burgdorferi invasiveness correlates with ospC genotype. To test this hypothesis and whether identical genotypes infect humans and small mammals in specific sites, B. burgdorferi ospC heterogeneity was tested among isolates from northern Maryland and southern Pennsylvania. Six culture-positive patients allowed collection of small animals from their properties, and spirochetes from animals trapped within 300 yards of each patient's home were isolated. 3' ospC sequences were compared to reference sequences. Of the 7 human and 15 mouse DNA templates that produced reliable sequences, all clustered with references into only four and seven distinct clades, respectively. A human and a mouse isolate with the same ospC were seen in only one locality, and five of six sites contained two or more B. burgdorferi ospC clones. Four invasive patient isolates and six small mammal isolates clustered with "noninvasive" reference ospC genotypes. A high degree of ospC diversity exists among B. burgdorferi isolates in Maryland and Pennsylvania, even in narrowly defined geographic localities. Dissemination in mice and humans by noninvasive ospC types contradicts the ospC invasiveness hypothesis. Alternative genetic markers for B. burgdorferi disseminated disease should be investigated.

Animals↗

Visualization of kinetochores in mammalian meiotic preparations and observations of argentophilic differences between mitotic and meiotic kinetochores.

Brief staining with AgNO3 was found to differentially stain the kinetochores of chromosomes from diakinesis - metaphase I and metaphase II nuclei of mammals. The results differ from those of Giemsa-stained or C-banded preparations as the silver-stained meiotic kinetochores are clearly distinguishable from both constitutive heterochromatin and euchromatin. Silver-staining is presented as a valuable method for the staining of meiotic material because it allows for the positive identification of centromeric position and orientation with respect to chiasmata. The nonargentophilic nature of the centromere (kinetochore) region of spermatogonial metaphase chromosomes in some species suggests a fundamental structural difference between mitotic and meiotic kinetochores.

Animals↗

Spotted fever group rickettsiae in immature and adult ticks (Acari: Ixodidae) from a focus of Rocky Mountain spotted fever in Connecticut.

Immature and adult ixodid ticks were collected during 1983 and 1984 in Newtown, Connecticut, an area endemic for Rocky Mountain spotted fever (RMSF), to determine prevalence of infection by spotted fever group (SFG) rickettsiae. Direct fluorescent-antibody (FA) staining revealed SFG organisms in 6 (1.8%) of 332 Dermacentor variabilis larvae, 5 (7.8%) of 64 D. variabilis nymphs, and in 2 (40%) of 5 Ixodes cookei nymphs removed from small- and medium-sized mammals. Hemolymph tests detected rickettsia-like organisms in 15 (8.8%) of 170 D. variabilis adults; 8 specimens retested by direct FA were negative. In contrast, hemocytes from 5 (8.6%) of 58 Ixodes texanus females contained organisms that stained positively in both hemolymph and direct FA tests. An indirect microimmunofluorescence test identified specific antibodies to Rickettsia rickettsii, the etiologic agent of RMSF, in serum samples from a chipmunk, raccoons, and white-footed mice. Results indicate that immature or adult ticks of at least three species may be involved in the maintenance and transmission of SFG rickettsiae at Newtown.

Animals↗

Differential effects of food restriction on pituitary-testicular function in mice.

The reproductive responses of two species of wild rodents, house mice and deer mice, were evaluated following a 30% reduction in food intake for 5 wk. These animal models were chosen as prototypes of other rodent species because each employs unique functional adjustments when confronted with reduced resources in their natural habitats. Modest inanition failed to alter pituitary-testicular function in house mice; neither spermatogenesis nor plasma concentrations of luteinizing hormone (LH) and testosterone were modified. In sharp distinction, deer mice exposed to restricted food intake showed significant reductions in plasma LH and testosterone and an accompanying loss in spermatogenesis. Reduced food intake also caused pronounced shifts in the temporal organization and amount of wheel-running activity in both animal models, albeit in a dichotomous fashion. House mice exhibited the same amount of wheel-running activity throughout inanition, but the diel periodicity of locomotor behavior was shifted from the dark to the light period. Deer mice, in comparison, significantly curtailed wheel-running activity during the dark hours but ran in precise phase relationship with the light-dark cycle. Taken together, our results establish that the male reproductive system and its supporting neuroendocrine and behavioral correlates can be disrupted by modest levels of food restriction in certain animal models.

Animals↗

Measurement of 2H2O by IR absorbance in doubly labeled H2O studies of energy expenditure.

The energy expenditure of animals in their natural surroundings can be determined by measuring the turnover in body water of isotopes of oxygen and hydrogen. We evaluated the use of infrared spectrophotometry for measuring 2H2O in small (20-microliters) water samples also labeled with 18O. For 2H2O over the enrichment range of 0.1-1 atom%, there was a linear relationship between infrared absorbance and 2H2O enrichment. 2H2O enrichments could be measured with a precision and accuracy of less than or equal to 1%, using this relationship. The presence of 18O in water samples in enrichments of up to 1 atom% had no significant effect on measurement of 2H2O by infrared absorbance. We measured the simultaneous turnover rates of 2H2O and 3H in mice and turtles also labeled with 18O. Our results validated the use of infrared absorbance in doubly labeled water measures of energy expenditure and indicated that the fractionation factors in vivo for 2H2O and 3H do not differ.

Algorithms↗

Effects of photoperiod and 2-deoxy-D-glucose-induced metabolic stress on immune function in female deer mice.

Nontropical rodents may experience large fluctuations in both food availability and energetic demands. The energy required for thermoregulation is high during the winter when energy availability is usually low. Winter conditions can induce a state of energetic stress that elevates circulating glucocorticoid levels and compromises immune function. Exposure to short days enhances immune function; the adaptive function of short-day enhancement of immune function may be to counteract the effects of stress-induced immunocompromise. To examine the role of energy availability in immune function, female deer mice were housed in either long (16:8-h light-dark cycle) or short (8:16-h light-dark cycle) days for 8 wk and then injected with either saline or 2-deoxy-D-glucose (2-DG), a glucose analog that inhibits cellular utilization of glucose and induces energetic stress. Long-day mice injected with 2-DG exhibited elevated corticosterone levels and reduced splenocyte proliferation compared with control mice. Short days buffered the animals against glucoprivation stress. Neither corticosterone levels nor splenocyte proliferation differed between 2-DG injected and control mice housed in short days. These data are consistent with the hypothesis that short days provide a buffer against metabolic stress.

Animals↗

Changes in serotonin levels, N-acetyltransferase activity, hydroxyindole-O-methyltransferase activity, and melatonin levels in the pineal gland of the Richardson's ground squirrel in relation to the light-dark cycle.

Pineal serotonin and melatonin levels and the activities of hydroxyindole-O-methyltransferase (HIOMT) and N-acetyltransferase (NAT) were studied over a 24-hour period in the pineal gland of the diurnally active Richardson's ground squirrel (Spermophilus richardsonii). Under alternating light-dark conditions (light:dark hours 14:10), pineal serotonin and melatonin levels exhibited a rhythm with high values occurring either during the day (serotonin) or during the night (melatonin). NAT activity was also markedly increased during darkness. HIOMT activity exhibited no 24-hour variation. Exposure of squirrels to constant light for 7 days exaggerated the serotonin rhythm, but obliterated the cycles of NAT and melatonin. Under constant darkness (for 7 days), the rhythms in serotonin, melatonin and NAT persisted, each having a period of about 24 h. In the second study, ground squirrels were exposed to light-dark cycles of either 8:16, 10:14 or 14:10. Under each of these photoperiodic environments, rhythms in pineal NAT and melatonin were apparent. Increasing the daily dark period from 10 to 14 h caused a prolongation of the elevated NAT and melatonin levels. However, a further prolongation of the daily dark period (to 16 h) did not further increase the duration of the rise in NAT and melatonin. The results show that continual light exposure (irradiance of 200 microW/cm2) for 7 days suppresses the pineal rhythms in both NAT activity and melatonin level in the Richardson's ground squirrel. Conversely, light exposure, rather than depressing the serotonin rhythm, actually exaggerates it. Constant darkness for 7 days has little influence on the 24-hour rhythms of either NAT or melatonin.(ABSTRACT TRUNCATED AT 250 WORDS)

Acetylserotonin O-Methyltransferase↗

Lithium chloride as an enhancing agent for metaphase preparations from rodent lymphocytes.

Circulating lymphocytes of many mammals, especially rodents, have proven refractory to PHA stimulation of mitotic activity. Addition of small amounts of lithium chloride to the culture medium removes the refractoriness and increases the yield of metaphase cells 7 fold. This simple procedure makes possible the routine use of circulating lymphocytes in rodent karyology.

Animals↗

Synapsis of a chromosomal pair heterozygous for a pericentric inversion and the presence of a heterochromatic short arm.

Analysis of meiotic pairing configurations in a deer mouse heterozygous for both a pericentric inversion and the presence of a heterochromatic short arm at chromosome 15 revealed straight-paired synaptonemal complexes with equal axial lengths in a majority of the pachytene nuclei. Nonhomologous pairing in this bivalent occurs by direct heterosynapsis of the inverted segments followed by synaptic adjustment of the heterochromatin heteromorphism.

Animals↗

Cryogenic preservation of mammalian testicular material for synaptonemal complex analysis.

Synaptonemal complex (SC) analyses have been used in mammalian cytogenetics to investigate the effects of chromosome heterozygosity on meiotic processes. To date, these analyses have been limited largely to the study of humans, livestock, or species which are easily maintained in the laboratory. The development of a method for the cryogenic preservation of testicular tissue for SC analyses allows for the application of this approach to field-oriented biological problems. This new method is described and data are presented which demonstrate its utility to mammalian cytogenetic studies.

Animals↗

Choroid plexitis in white-tailed deer (Odocoileus virginianus) in southern New York State.

Brains, spinal cords, nerve roots, nerves and muscle tissues were removed from deer in southern New York State and examined for histologic evidence of infection by the causative agent of Lyme disease, Borrelia burgdorferi. There was no histologic evidence of this infection and only four of 26 deer had serologic evidence of past infection despite the fact that all were parasitized by the tick vector, Ixodes dammini. Of these ticks, 21% were carrying B. burgdorferi. In contrast, most of the deer had choroid plexitis. All but one of 48 deer tested were infected with Trypanosoma cervi, 20 of 24 deer had sarcocystis in skeletal muscles and two had dural lesions probably due to the nematode Pneumostrongylus tenuis. The causal relationship between choroid plexitis and trypanosomiasis is discussed.

Animals↗

Voluntary running in deer mice: speed, distance, energy costs and temperature effects.

The energetics of terrestrial locomotion are of considerable interest to ecologists and physiologists, but nearly all of our current knowledge comes from animals undergoing forced exercise. To explore patterns of energy use and behavior during voluntary exercise, we developed methods allowing nearly continuous measurements of metabolic rates in freely behaving small mammals, with high temporal resolution over periods of several days. We used this approach to examine relationships between ambient temperature (Ta), locomotor behavior and energy costs in the deer mouse, a small mammal that routinely encounters a large range of temperatures in its natural habitat. We tested for individual consistency in running behavior and metabolic traits, and determined how locomotor costs vary with speed and Ta. Because of the importance of thermoregulatory costs in small mammals, we checked for substitution of exercise heat for thermostatic heat production at Ta below the thermal neutral zone and determined the fraction of the daily energy budget comprising exercise costs. Locomotor behavior was highly variable among individuals but had high repeatability, at least over short intervals. We found few temperature-related changes in speed or distance run, but Ta strongly affected energy costs. Partial substitution of exercise heat for thermogenic heat occurred at low Ta. This reduced energy expenditure during low-temperature running by 23-37%, but running costs comprised a fairly minor fraction of the energy budget, so the daily energy savings via substitution were much smaller. Deer mice did not adjust running speed to maximize metabolic economy, as they seldom used the high speeds that provide the lowest cost of transport. The highest voluntary speeds (4-5 km h(-1)) were almost always below the predicted maximal aerobic speed, and were much less than the species' maximal sprint speed. Maximum voluntarily attained rates of oxygen consumption (VO2) were highest at low Ta, but rarely approached maximal VO2 during forced treadmill exercise. Mean respiratory exchange ratios coincident with maximal voluntary VO2 increased slightly as Ta declined, but were always below 1.0 (another indication that metabolic rate was less than the aerobic maximum). Individuals with high running performance (cumulative distance and running time) had high resting metabolism, which suggests a cost of having high capacity or propensity for activity.

Animals↗