Long-term behavioral effects of social separation during early life in a social mammal, Octodon degus.

Social separation is thought to induce a strong stress response in social juvenile mammals, but little is known about how this response might vary throughout the development. The present study examines the long-term effects of early-life stress (ELS) induced by social separation on individual behaviors later in life using the social and precocious species Octodon degus. Four experimental groups were established a positive control group of mothers and siblings from six litters comprised the socially housed (SH) group, while pups from seven litters were randomly assigned to three treatments: pups experiencing no separation (NS) treatment while their siblings did; repeated bouts of consecutive separation (CS); intermittent separation (IS). We analyzed the effects of separation treatment on the frequency and duration of freezing, rearing and grooming behaviors. ELS was correlated with higher hyperactivity, and hyperactivity increased with more frequent separation. However, the behavioral trend of the NS group changed to hyperactive in long-term observation. The findings suggest that the NS group was indirectly affected by ELS. In addition, suggesting ELS acts to converge an individual's behavioral tendencies in a certain direction.

Effects of aging and delivery order on the breeding capacity of F344/N rats.

Reproductive performance affects the efficiency of the production of animals. Therefore, knowing the reproductive properties of each species or strain of animal is important for proper management of breeding stocks. To elucidate the reproductive properties of female F344/N rats, frequently used for longevity and gerontology research, we monitored the breeding duration, number of deliveries, litter size and weaning rate of their pups. The first delivery was observed at 2.9 ± 0.1 (mean ± standard deviation) months of age (M) and the last delivery was at 15.1 ± 1.8 M. The number of deliveries was 10.4 ± 2.8 (range of 7-16) within the life span of the mother (24.7 ± 5.6 M). The litter sizes at the third (10.0 ± 3.0) or fourth (10.3 ± 2.7) deliveries were higher than for other deliveries. The breeding outcome declined after the fifth delivery yielding reduced litter size or weaning rate, number of delivered mother. These results suggests that though female F344/N rats are able to deliver more than 10 times, the reproductive performance lowered after fifth delivery.

Interspecific variation in wildlife responses to cattle, swine and chicken feed in the forests surrounding poultry farms.

To strengthen farm biosecurity, wildlife behaviors around livestock environments require significant attention. Livestock feed is considered one of essential factors that attract wildlife to the livestock environment. We experimentally studied wildlife response to cattle, swine, and chicken concentrated feeds in the forests surrounding poultry farms. In 14 feeding sites, four feed conditions were established: without feed (control); cattle feed; chicken feed; and swine feed. Wildlife behaviors at each feed point were monitored using infrared cameras. In 3,175 videos, 10 mammals were photographed on 10 or more occasions. Wildlife more frequently appeared at the points with feed than without feed. In addition, the number of videos that captured foraging or interest behaviors was largest for swine feed, followed by chicken feed, then cattle feed. There was a large difference among wildlife in their response to livestock feeds, although each species did not have a strong preference for a specific feed. Livestock feeds invite frequent visits by high and moderate response groups, especially omnivores and carnivores with omnivorous tendencies. Therefore, to protect against such wildlife intrusion, leftover feed and feed storage must be properly managed. This study also suggests that livestock feeds may not cause intrusions by rare response group species; hence, if their intrusions occur, they may be due to factors other than livestock feed. The study situation can partly reflect actual feed-stealing situations. The results will contribute to consider the properly management to protect livestock environments from wildlife intrusions.

Antibiotic-resistant Escherichia coli isolated from dairy cows and their surrounding environment on a livestock farm practicing prudent antimicrobial use.

On a livestock farm where antimicrobial administration and its history had been managed for prudent use of antimicrobials, we surveyed antibiotic-resistant Escherichia coli strains isolated from cow feces and the surrounding environment (i.e., rat and crow feces, and water samples from a drainage pit and wastewater processing tank) every month for 1 year. Two strains (1.7%) in cow feces were resistant to tetracycline, whereas all other strains were susceptible to all other antimicrobials. Among 136 strains isolated from cows and wild animals, only one ampicillin-resistant strain was identified. The antibiotic resistance rate in the drainage from the barn was 8.3% (10/120), and all strains showed susceptibility for 8 months of the year. Tetracycline resistance was common in all resistant strains isolated from animal feces and water samples; all tetracycline-resistant strains carried tetA. These results strongly support the proper use and management of antibiotics on farms to minimize the outbreak and spread of antibiotic-resistant bacteria.

Individual variation of daily torpor and body mass change during winter in the large Japanese field mouse (Apodemus speciosus).

Daily torpor is a strategy used by some overwintering small endotherms to aid in energy conservation. However, the pattern of torpor varies among individuals within species and populations, even under the same environmental conditions, with significant implications for survival rate and reproductive success. Body mass is one factor that may influence this variation, especially in some small mammals that accumulate fat stores prior to overwintering. However, to our knowledge there has been no previous study examining the detailed relationships between torpor expression and body mass change in small mammals that hoard food as an energy resource during winter. The large Japanese field mouse, Apodemus speciosus, whose winter survival strategy depends on food caches instead of fat stores, displays daily torpor under artificial winter conditions (short-day photoperiod and cold). The present study clarifies the characteristics and patterns of daily torpor and body mass change in this species in the laboratory. Although expression of daily torpor was facilitated progressively as in other species, the observed patterns of torpor expression and body mass change showed considerable individual variation. Moreover, there was no obvious correlation between body mass and daily torpor expression. Therefore, it is suggested that in A. speciosus body mass may not contribute to individual variation of daily torpor during winter. Daily torpor during winter may be adjusted by not only mechanisms common to other small mammals, but also species-specific factors relating to the external or internal reserves of energy in small mammals.

Individual differences in torpor expression in adult mice are related to relative birth mass.

Daily torpor is a physiological adaptation in small mammals and birds, characterised by drastic reductions in metabolism and body temperature. Energy-constraining conditions, such as cold and starvation, are known to cause the expression of daily torpor. However, the reason for high degrees of inter- and intra-individual variation in torpor expression (TE) in similar situations is not clear. As littermates of altricial animals are exposed to an uneven allocation of maternal resources from conception to weaning, we tested whether early nutritional experiences have long-term effects on TE in adults. We used full-sibling littermates of laboratory mice that as adults were starved overnight to induce torpor. We measured body mass from birth until adulthood as an indicator of nutritional status, and calculated the relative body mass (RBM) as an indicator of the difference in nutritional status within a litter. After maturation, we subjected mice to five repeated torpor induction trials involving 24 h of fasting and 5 days of recovery. Half of the female mice displayed great individual variation in TE whereas male mice rarely exhibited daily torpor. In females, RBM at birth influenced TE, irrespective of body mass in adulthood; thus, female mice born with low RBMs displayed high TE in adulthood. In conclusion, we provide evidence that TE in mice differs among littermates, and that this variation is linked closely to heterogeneous nutritional experiences during the fetal period.

Chromosomal Aberrations in Large Japanese Field Mice (Apodemus speciosus) Captured near Fukushima Dai-ichi Nuclear Power Plant.

Since the Fukushima Dai-ichi Nuclear Power Plant accident, radiation effects on nonhuman biota in the contaminated areas have been a major concern. Here, we analyzed the frequencies of chromosomal aberrations (translocations and dicentrics) in the splenic lymphocytes of large Japanese field mice (Apodemus speciosus) inhabiting Fukushima Prefecture. A. speciosus chromosomes 1, 2, and 5 were flow-sorted in order to develop A. speciosus chromosome-specific painting probes, and FISH (fluorescence in situ hybridization) was performed using these painting probes to detect the translocations and dicentrics. The average frequency of the translocations and dicentrics per cell in the heavily contaminated area was significantly higher than the frequencies in the case of the noncontaminated control area and the slightly and moderately contaminated areas, and this aberration frequency in individual mice tended to roughly increase with the estimated dose rates and accumulated doses. In all four sampling areas, the proportion of aberrations occurring in chromosome 2 was approximately >3 times higher than that in chromosomes 1 and 5, which suggests that A. speciosus chromosome 2 harbors a fragile site that is highly sensitive to chromosome breaks induced by cellular stress such as DNA replication. The elevated frequency of chromosomal aberrations in A. speciosus potentially resulting from the presence of a fragile site in chromosome 2 might make it challenging to observe the mild effect of chronic low-dose-rate irradiation on the induction of chromosomal aberrations in A. speciosus inhabiting the contaminated areas of Fukushima.

Chromosomal Aberrations in Wild Mice Captured in Areas Differentially Contaminated by the Fukushima Dai-Ichi Nuclear Power Plant Accident.

Following the Fukushima Dai-ichi Nuclear Power Plant accident, radiation effects on nonhuman biota in the contaminated areas have been a great concern. The induction of chromosomal aberrations in splenic lymphocytes of small Japanese field mice (Apodemus argenteus) and house mice (Mus musculus) inhabiting Fukushima Prefecture was investigated. In mice inhabiting the slightly contaminated area, the average frequency of dicentric chromosomes was similar to that seen in mice inhabiting a noncontaminated control area. In contrast, mice inhabiting the moderately and heavily contaminated areas showed a significant increase in the average frequencies of dicentric chromosomes. Total absorbed dose rate was estimated to be approximately 1 mGy d(-1) and 3 mGy d(-1) in the moderately and heavily contaminated areas, respectively. Chromosomal aberrations tended to roughly increase with dose rate. Although theoretically, the frequency of chromosomal aberrations was considered proportional to the absorbed dose, chromosomal aberrations in old mice (estimated median age 300 days) did not increase with radiation dose at the same rate as that observed in young mice (estimated median age 105 days).

Magnitude of food overabundance affects expression of daily torpor.

Many small mammal species use torpor as a strategy for reducing energy expenditure in winter. Some rodent hibernators also hoard food to provide reserves of energy, and individuals with large hoards express less torpor than those with smaller reserves. These facts imply that animals can recognize levels of food availability, but where food is very plentiful, it is unclear whether torpor expression is affected by temporal changes in the extent of food overabundance. Moreover, the relationship between daily torpor and excess food availability has not been clearly established. The large Japanese field mouse Apodemus speciosus caches food for use as a winter energy resource and exhibits daily torpor under artificial winter conditions. The present study examined whether individuals exposed to different magnitudes of overabundant food exhibited differences in expression of daily torpor, and secondly whether torpor expression varied in response to changes in the overall quantity of overabundant food. It was observed that while absolute quantities of overabundant food did not appear to affect daily torpor expression, the mice did respond to changes in food availability, even when food remained overabundant. This suggests that the mice respond to fluctuations in food availability, even where these changes do not place any constraint on energy budgets. Thus recognition of changing food availability cannot be a purely physiological response to shortage or plenty, and may contribute to predictions of future energy availability. The expression of torpor was inhibited in response to increasing food availability, and the mice used shallower torpor when food availability increased to superabundance. These findings suggest that daily torpor may be regulated not only physiologically in response to energy constraints but also psychologically, via recognition of food availability.

The effects of extracellular ATP and its receptor antagonists on pig oocytes during in vitro maturation.

We measured the ATP concentrations in the porcine follicular fluid derived from three sizes of follicles (small: 6 mm in diameter). Then, the effects of pre-treatment (100 µM each for 30 min before maturation) with antagonists for extracellular ATP receptor P2X or P2Y on the nuclear maturation rate of cumulus-cell-enclosed (COs) or -denuded oocytes (DOs) up to the preovulatory stage in the presence or absence of 20 nM ATP (a similar concentration to that of medium-sized follicle fluid) were investigated. The antagonists used were pyridoxalphosphate-6-azophenyl-2',4'-disulfonic acid (PPADS) or reactive blue 2 (RB2), for extracellular ATP receptor P2X and P2Y, respectively. In addition, the embryonic development rates of COs pre-treated with RB2 were also evaluated. It was found that when the follicular sizes increased, the ATP concentrations significantly decreased (P < 0.05). No differences were observed in the nuclear maturation rates among all COs, regardless of pre-treatment with (+) or without (-) PPADS and in the presence (+) or absence (-) of ATP during maturation. In contrast, the nuclear maturation rate of the COs, but not DOs, in the ATP(-) RB2(+) group was significantly lower (P < 0.05) than that of the ATP(-) RB2(-) and ATP(+)RB2(-) groups. The pronuclear formation and blastocyst formation rates by parthenogenetic activation in the ATP(-) RB2(+) and ATP(+) RB2(+) groups were significantly lower (P < 0.05) than those in the ATP(-) RB2(-) group. In conclusion, it is suggested that the nuclear maturation of porcine oocytes may be influenced by the ATP receptor P2Y present in the cumulus cells.

Huddling facilitates expression of daily torpor in the large Japanese field mouse Apodemus speciosus.

Small endotherms employ multiple adaptations to maintain energy balance in winter, including spontaneous daily torpor and simultaneous huddling. The relationships between these adaptations have been discussed in several previous studies, but it has not been well-established if huddling actually affects the expression of torpor in small endotherms. We examine whether and how huddling affects the expression of torpor in the large Japanese field mouse Apodemus speciosus, which is known to become torpid under artificial winter conditions. The mice were found to adjust expression of torpor in response to the number of cage mates. Torpor frequency and minimum torpid body temperature were both significantly elevated when the number of cage mates was increased, but there was no significant change in torpor bout length. Rewarming rate on arousal was lower when the number of cage mates was increased, suggesting reduction in endogenous rewarming due to exogenous passive rewarming. Food consumption per mouse decreased significantly with increasing number of cage mates. Thus, our study demonstrates that social thermoregulatory behaviors such as huddling can facilitate expression of spontaneous daily torpor in small rodents. These findings suggest that energy constraints, such as ambient temperature and food availability may not be the only modulating factors on the expression of daily torpor.

Populations of follicles in F344/N rats during aging.

Follicular populations were investigated in female F344/N rats to better understand the aging process of the rat ovary. Ovaries dissected at various ages (spanning 1-36 months old) were submitted for histological examination. The total number of primordial, growing (primary and secondary), tertiary, and atretic follicles as well as corpora lutea (CL) were counted in hematoxylin-eosin- and azocarmine-aniline-blue-stained ovarian sections. The number of healthy follicles including primordial, growing and tertiary follicles decreased rapidly between the first and third months and gradually thereafter. CL were found in 3-month-old rats, and their number remained unchanged until 18 months of age, at which point it decreased. The number of atretic follicles started to increase in rats older than 18 months, which corresponded to the cessation of estrous cyclicity. Several healthy follicles and CL were observed even in 36-month-old rats.

A distinct role of the queen in coordinated workload and soil distribution in eusocial naked mole-rats.

We investigated how group members achieve collective decision-making, by considering individual intrinsic behavioural rules and behavioural mechanisms for maintaining social integration. Using a simulated burrow environment, we investigated the behavioural rules of coordinated workload for soil distribution in a eusocial mammal, the naked mole-rat (Heterocephalus glaber). We tested two predictions regarding a distinct role of the queen, a socially dominant individual in the caste system: the presence of a queen would increase the workload of other caste individuals, and the cues by a queen would affect the soil distribution. In experiment 1, we placed four individuals of various castes from the same colony into an experimental burrow. Workers exhibited the highest frequency of workload compared to other castes. The presence of a queen activated the workload by other individuals. Individuals showed a consistent workload in a particular direction so as to bias the soil distribution. These results suggest that individuals have a consensus on soil distribution and that the queen plays a distinct role. In experiment 2, we placed the odour of a queen in one of four cells and observed its effect on other individuals' workload and soil distribution. Relative to other cells, individuals frequently dug in the queen cell so the amount of soil in the queen cell decreased. These results suggest that queen odour is an important cue in coordinated workload and soil distribution in this species.