Small Litters Research Articles

Effect of Endothelial Nitric Oxide Synthase Genotype on Outcomes after Experimental Thromboembolic Stroke

Acute ischemic stroke is the one of leading causes of morbidity and mortality in the U.S. African Americans have higher risk of stroke incidence and death that is associated with endothelial dysfunction and reduced bioavailability of nitric oxide. We propose that endothelial nitric oxide synthase knockout (eNOS−/−) or heterozygote (eNOS+/−) mice may be a suitable model of African American stroke.MethodsThe eNOS−/− and C57BL/6J mice were crossed to obtain eNOS+/−MAT and eNOS+/−PAT offspring (30±3 weeks; 13 mice/group). Embolic stroke was initiated by injection of a thrombus to the right middle cerebral artery. Cerebral blood flow was recorded to confirm occlusion. At 24 hours, the neurological status and brains were analyzed.ResultseNOS−/− mice were characterized by small litter size and high pup mortality. The final body weight was similar among eNOS+/−MAT, eNOS+/−PAT, and eNOS−/− mice, but significantly lower than in wild type mice (28.0±1.8 vs. 31.4±1.9; p<0.001). Cerebral blood flow, infarct size, and neurological scores were comparable in all groups. However, all nitric oxide knockout mice had markedly increased mortality compared with wild type mice (46% vs. 23%).ConclusioneNOS−/− or eNOS+/− mice may be used as a model of the cerebrovascular complications associated with abnormalities in nitric oxide signaling as seen in African Americans.Research support: NIH‐R21 award and GHSU start‐up to I.Y.S.

An overview on the role of Hexanchiformes in marine ecosystems: biology, ecology and conservation status of a primitive order of modern sharks

The large size, high trophic level and wide distribution of Hexanchiformes (cow and frilled sharks) should position this order as important apex predators in coastal and deep-water ecosystems. This review synthesizes available information on Hexanchiformes, including information not yet published, with the purpose of evaluating their conservation status and assessing their ecological roles in the dynamics of marine ecosystems. Comprising six species, this group has a wide global distribution, with members occurring from shallow coastal areas to depths of c. 2500 m. The limited information available on their reproductive biology suggests that they could be vulnerable to overexploitation (e.g. small litter sizes for most species and suspected long gestation periods). Most of the fishing pressure exerted on Hexanchiformes is in the form of commercial by-catch or recreational fishing. Comprehensive stock and impact assessments are unavailable for most species in most regions due to limited information on life history and catch and abundance time series. When hexanchiform species have been commercially harvested, however, they have been unable to sustain targeted fisheries for long periods. The potentially high vulnerability to intense fishing pressure warrants a conservative exploitation of this order until thorough quantitative assessments are conducted. At least some species have been shown to be significant apex predators in the systems they inhabit. Should Hexanchiformes be removed from coastal and deep-water systems, the lack of sympatric shark species that share the same resources suggests no other species would be capable of fulfilling their apex predator role in the short term. This has potential ecosystem consequences such as meso-predator release or trophic cascades. This review proposes some hypotheses on the ecology of Hexanchiformes and their role in ecosystem dynamics, highlighting the areas where critical information is required to stimulate research directions.

Relationships among steroid hormone levels in newborn piglets, birth weight, placental weight, vitality of offspring and litter size

Blood estrone sulfate (E(1) S), estrone (E(1) ), estradiol (E(2) ) and progesterone (P(4) ) in newborn piglets were measured to clarify the relationships among birth and placental weight, vitality of offspring and litter size. First, the association between vital status (normal, weak and stillborn) from 165 newborn piglets of 18 litters and steroid concentrations; second, steroid concentrations from 152 newborn normal piglets and litter size; and third, steroid content in fetal placenta from 50 newborn normal piglets of six litters and litter size, were investigated. In the normal group, the birth and placental weight were significantly higher than those in the other groups. Blood E(1) S levels in the stillborn group were significantly lower, whereas E(1) , E(2) and P(4) were significantly higher compared to the normal group. Blood and placental E(1) S levels in the small litter group were significantly higher than those in the other groups. However, there was no significant difference among the three litter size groups in the levels of steroid hormones in maternal blood. These results indicate that vitality of newborn piglets is related to E(1) S concentration of neonate, to birth weight and placental weight. However, steroid hormone concentrations of newborn piglets were greatly affected by the number of littermates.

Maternal behavior of the Eurasian lynx Lynx lynx L. during the early postnatal ontogeny of its cubs

The dynamics of the major elements of maternal behavior of Eurasian lynx females during the first month of life of their cubs and their association with litter parameters (number of kittens, sex, and mass) have been traced. By the end of the first month, the amount of time spent by the female outside of the den significantly increases. An association between the litter size and maternal behavior has been found. Females rearing small litters spend more time outside of their den; they also spend more time on allogrooming of each separate kitten than females with large litters. Concerning allogrooming duration, a preference for male kittens by lynx females has been noticed in the third week.

Anogenital distance as a predictor of attractiveness, litter size and sex ratio of rabbit does

In the case of some mammalian species, females with larger anogenital distance (AGD) have smaller litters, where the sex ratio is male-biased. The first aim of the present study was to test whether this phenomenon exists in the Lagomorph group, especially in rabbits. The results revealed that does with large AGD have significantly smaller and lighter litters with a male biased sex ratio; with fewer females but not more males.The second aim of the present study was to test whether males differentiate between females differing in their AGD and are thus capable of choosing the more fertile and fecund does. Males showed a stronger response to the chin marks of females with small AGD than to the marks of females with large AGD. Our results suggest that variation in the prenatal hormonal environment, reflected through variation in AGD, could have long-term consequences on mate choice and population dynamics.

Reproduction of the root vole (Microtus oeconomus) at the edge of its distribution range

Data on the reproduction of the root vole (Microtus oeconomus) in Lithuania, the margin of its geographic range, are presented in the context of litter size analysis. In Lithuania, the breeding season lasts from April until the end of October. The average litter size was 5.93 ± 0.11 (2-14), decreasing from a mean of 8.0 embryos in June to 4.3 in October, with 2-3 litters per year. The average body mass of breeding females was 40.4 ± 0.6 (16.5-77.0) g. However, 6.25% of females start breeding with a body mass of under 25 g. Litter size was found to increase from south to north in the geographical distribution range of the species and is described by the regression Litter size = 0.11147 x latitude - 0.2646 (R^2 = 0.79). The small litter size and the comparatively small number of litters in the Lithuanian population indicate that reproduction is similar to that of other populations in the southern part of the geographical range of the species.

Growth and mortality of SIKA suckling rabbits in Slovenia

The aim of the study was to estimate some effects on mortality and growth in rabbits. The study was carried out in Slovenian SIKA sire line. In total, 1028 kits of 80 does were included. In the analyses the effects of parity, number of liveborn kits, number of teats, and season of kits birth were analysed. Birth weight was included in the model for mortality, while age was included in the model for growth. Parity, litter size, teat number and season affected the body weights. Body weight varied also according to age. Mortality has changes by litter size, season and initial weight. An average litter size was 8.77 kits born, 8.15 kits born alive and 7.00 kits weaned. The birth to weaning mortality was 14.4% and mortality has steadily declined with age. Average weight up to age 3 days was 72 g and at weaning 959 g. Kits in smaller litters and kits from does with more teats had a higher growth rate. Also kits grow faster in the colder months.

Neonatal Overfeeding Induced by Small Litter Rearing Causes Altered Glucocorticoid Metabolism in Rats

Elevated glucocorticoid (GC) activity may be involved in the development of the metabolic syndrome. Tissue GC exposure is determined by the tissue-specific GC-activating enzyme 11β-hydroxysteriod dehydrogenase type 1 (11β-HSD1) and the GC-inactivating enzyme 5α-reductase type 1 (5αR1), as well as 5β-reductase (5βR). Our aim was to study the effects of neonatal overfeeding induced by small litter rearing on the expression of GC-regulating enzymes in adipose tissue and/or liver and on obesity-related metabolic disturbances during development. Male Sprague-Dawley rat pup litters were adjusted to litter sizes of three (small litters, SL) or ten (normal litters, NL) on postnatal day 3 and then given standard chow from postnatal week 3 onward (W3). Small litter rearing induced obesity, hyperinsulinemia, and higher circulating corticosterone in adults. 11β-HSD1 expression and enzyme activity in retroperitoneal, but not in epididymal, adipose tissue increased with postnatal time and peaked at W5/W6 in both groups before declining. From W8, 11β-HSD1 expression and enzyme activity levels in retroperitoneal fat persisted at significantly higher levels in SL compared to NL rats. Hepatic 11β-HSD1 enzyme activity in SL rats was elevated from W3 to W16 compared to NL rats. Hepatic 5αR1 and 5βR expression was higher in SL compared to NL rats after weaning until W6, whereupon expression decreased in the SL rats and remained similar to that in NL rats. In conclusion, small litter rearing in rats induced peripheral tissue-specific alterations in 11β-HSD1 expression and activity and 5αR1 and 5βR expression during puberty, which could contribute to elevated tissue-specific GC exposure and aggravate the development of metabolic dysregulation in adults.

Litter sizes of brown bears in the Central Alborz Protected Area, Iran

Abstract Although smaller than 4,000 km2, the Central Alborz Protected Area (CAPA) is one of the main habitats of brown bear (Ursus arctos) in Iran. During August 2005 to September 2009, we gathered data through direct observations of bears, identifying individual bears by means of age, sex, color, and behavior. We observed bears on 115 occasions. Mean size of cub litters was 2.00 (SE = 0.20, n = 13) and varied from 1 to 3. We speculate that low occurrence of meat in food items of the bears in the area explains this relatively small litter size. We hypothesize that the north-central portion of the Alborz Protected Area is a female core area which supports surrounding sink populations and needs to be protected more effectively.

Stress-induced sex ratios in ground squirrels: support for a mechanistic hypothesis

Systematic deviations in sex ratio may be adaptive in the face of the prevailing environmental or physiological cues experienced by the mother; yet some theoretical and mechanistic hypotheses remain at odds and are rarely examined noninvasively under natural conditions. Conventional interpretations of the Trivers and Willard (Trivers RL, Willard DE. 1973. Natural selection of parental ability to vary the sex ratio of offspring. Science 179:90–92) hypothesis (TWH) predict that higher stress should be associated with female-biased litters; yet, several mechanistic hypotheses predict the opposite. We tested the predictions of the TWH and 2 stress-related mechanistic hypotheses in a free-living polygynandrous sciurid, Richardson’s ground squirrel, Urocitellus richardsonii. We examined the relationship between litter sex ratio and indicators of maternal condition and investment. These included litter size, juvenile mass at emergence, as well as maternal age, changes in maternal body mass, and maternal fecal glucocorticoid (cortisol and corticosterone) levels during gestation, lactation, and post-weaning. Males born of small litters were significantly heavier at emergence than males from larger litters, which were female biased, whereas females showed no significant change in mass with litter size. Mothers with higher fecal cortisol levels during the gestation period (but not during lactation or post-weaning) were more likely to produce male-biased litters, whereas females producing larger litters showed significantly higher cortisol levels during lactation (but not gestation or post-weaning) than mothers producing smaller litters. Our results provide support for both the proximate glucose-mediated mechanistic model of Cameron (Cameron EZ. 2004. Facultative adjustment of mammalian sex ratios in support of the Trivers-Willard hypothesis: evidence for a mechanism. Proc R Soc Ser B Biol Sci. 271:1723–1728) and the broad-sense TWH, where social and life-history characteristics play an integral role.

Optimizing temperament through litter size in short‐lived, iteroparous mammals in seasonal environments

A number of short-lived, iteroparous animal species have small broods in the early breeding season and larger broods in later breeding season. Brood size affects not only offspring size, but as recent results suggest, may also affect offspring's temperament, hormonal status, and aggression as adults. Most populations of short-lived, iteroparous mammals fluctuate predictably over the season, with low densities in winter, increasing densities in summer and a population peak in late summer followed by a population breakdown. If animals live only through parts of the season, possibly such differences in density and hence also in social environments among seasons require different personality types to increase individual fitness. We present data on behavior of European rabbits from a field enclosure study. These data clearly show that aggressiveness is higher in young from smaller litters than in young from larger litters, and smaller litters are usually born during the early breeding season. Moreover, our data suggest that behavioral types of the young rabbits are stable over time, at least during their subadult life. We suggest, that changes in mean litter size over the course of the breeding season may not only be a product of mothers' age or food availability, but may also have an adaptive function by preparing offspring characteristics for adulthood in a social environment undergoing predictable density changes within the season.

MyD88 Deficiency Ameliorates β-Amyloidosis in an Animal Model of Alzheimer's Disease

The accumulation of β-amyloid protein (Aβ) in the brain is thought to be a primary etiologic event in Alzheimer's disease (AD). Fibrillar Aβ plaques, a hallmark of AD abnormality, are closely associated with activated microglia. Activated microglia have contradictory roles in the pathogenesis of AD, being either neuroprotective (by clearing harmful Aβ and repairing damaged tissues) or neurotoxic (by producing proinflammatory cytokines and reactive oxygen species). Aβ aggregates can activate microglia by interacting with multiple toll-like receptors (TLRs), the pattern-recognition receptors of the innate immune system. Because the adapter protein MyD88 is essential for the downstream signaling of all TLRs, except TLR3, we investigated the effects of MyD88 deficiency (MyD88(-/-)) on Aβ accumulation and microglial activation in an AD mouse model. MyD88 deficiency decreased Aβ load and microglial activation in the brain. The decrease in Aβ load in an MyD88(-/-) AD mouse model was associated with increased and decreased protein expression of apolipoprotein E (apoE) and CX3CR1, respectively, compared with that in an MyD88 wild-type AD mouse model. These results suggest that MyD88 deficiency may reduce Aβ load by enhancing the phagocytic capability of microglia through fractalkine (the ligand of CX3CR1) signaling and by promoting apoE-mediated clearance of Aβ from the brain. These findings also suggest that chronic inflammatory responses induced by Aβ accumulation via the MyD88-dependent signaling pathway exacerbate β-amyloidosis in AD.

Postnatal early overnutrition dysregulates the intrarenal renin–angiotensin system and extracellular matrix-linked molecules in juvenile male rats

Overnutrition during the perinatal period has been associated with susceptibility to obesity and related comorbidities. We examined the effects of postnatal early overnutrition on the development of juvenile obesity and the associated renal pathophysiological changes. Three or 10 pups per mother from rat pup litters were assigned to either the overnutrition or control groups during the first 21 days of life. The effects of overfeeding were measured at 28 days. The smaller male litter pups were heavier than the controls between 4 and 28 days after birth (P<.05). By 28 days of age, the kidney weight per body weight ratio decreased in the small litter group (P<.05). Circulating leptin levels increased in the small litter rats (P<.05). Overnutrition had no effect on renal cell proliferation, apoptosis, macrophages and glomerulosclerosis. In the immunoblots and immunohistochemistry, renin and angiotensin II type (AT) 2 receptor expression increased in the overfed rats (P<.05). By contrast, the plasminogen activator inhibitor (PAI)-1 and matrix metalloproteinase (MMP)-9 expression decreased in the overnutrition group (P<.05). The AT 1 receptor, tissue inhibitor of MMP-1, monocyte chemoattractant protein-1, tumor necrosis factor-α, osteopontin and adiponectin expression was not changed. Our data showed that postnatal early overfeeding led to hyperleptinemia, juvenile obesity and the acquired reset of renal maturation. Up-regulation of renin and AT2 and down-regulation of PAI-1 and MMP-9 might contribute to abnormal programming of renal growth in rats exposed to postnatal early overnutrition.

Early Metabolic Programming of Puberty Onset: Impact of Changes in Postnatal Feeding and Rearing Conditions on the Timing of Puberty and Development of the Hypothalamic Kisspeptin System

Kiss1 neurons have recently emerged as a putative conduit for the metabolic gating of reproduction, with leptin being a regulator of hypothalamic Kiss1 expression. Early perturbations of the nutritional status are known to predispose to different metabolic disorders later in life and to alter the timing of puberty; however, the potential underlying mechanisms remain poorly defined. Here we report how changes in the pattern of postnatal feeding affect the onset of puberty and evaluate key hormonal and neuropeptide [Kiss1/kisspeptin (Kp)] alterations linked to these early nutritional manipulations. Female rats were raised in litters of different sizes: small (four pups per dam: overfeeding), normal (12 pups per dam), and large litters (20 pups per litter: underfeeding). Postnatal overfeeding resulted in persistently increased body weight and earlier age of vaginal opening, as an external sign of puberty, together with higher levels of leptin and hypothalamic Kiss1 mRNA. Conversely, postnatal underfeeding caused a persistent reduction in body weight, lower ovarian and uterus weights, and delayed vaginal opening, changes that were paralleled by a decrease in leptin and Kiss1 mRNA levels. Kisspeptin-52 immunoreactivity (Kp-IR) in the hypothalamus displayed similar patterns, with lower numbers of Kp-IR neurons in the arcuate nucleus of postnatally underfed animals, and a trend for increased Kp-positive fibers in the periventricular area of early overfed rats. Yet, gonadotropin responses to Kp at puberty were similar in all groups, except for enhanced responsiveness to low doses of Kp-10 in postnatally underfed rats. In conclusion, our data document that the timing of puberty is sensitive to both overfeeding and subnutrition during early (postnatal) periods and suggest that alterations in hypothalamic expression of Kiss1/kisspeptin may underlie at least part of such programming phenomenon.

Developmental overfeeding alters hypothalamic neuropeptide mRNA levels and response to a high-fat diet in adult mice

It has been suggested that nutritional manipulations during the first weeks of life can alter the development of the hypothalamic circuits involved in energy homeostasis. We studied the expression of a large number of the hypothalamic neuropeptide mRNAs that control body weight in mice that were overfed during breastfeeding (mice grown in a small litter, SL) and/or during adolescence (adolescent mice fed a high-fat diet, AHF). We also investigated possible alterations in mRNA levels after 50 days of a high-fat diet (high-fat challenge, CHF) at 19 weeks of age. Both SL and AHF conditions caused overweight during the period of developmental overfeeding. During adulthood, all of the mouse groups fed a CHF significantly gained weight in comparison with mice fed a low-fat diet, but the mice that had undergone both breast and adolescent overfeeding (SL-AHF-CHF mice) gained significantly more weight than the control CHF mice. Of the ten neuropeptide mRNAs studied, only neuropeptide Y (NPY) expression was decreased in all of the groups of developmentally overfed adult mice, but CHF during adulthood by itself induced a decrease in NPY, agouti-related protein (AgRP) and orexin (Orx) mRNA levels. Moreover, in the developmentally overfed CHF mice NPY, AgRP, galanin (GAL) and galanin-like peptide (GalP) mRNA levels significantly decreased in comparison with the control CHF mice. These results show that, during adulthood, hypothalamic neuropeptide systems are altered (NPY) and/or abnormally respond to a high-fat diet (NPY, AgRP, GAL and GalP) in mice overfed during critical developmental periods.

Reproductive Ecology of American Black Bears in the Alleghany Mountains of Virginia, USA

AbstractWe studied reproduction of American black bears (Ursus americanus) in western Virginia from 1994 to 2003. We handled 326 ≥2‐year‐old female black bears 672 times during summer trapping and followed 176 of these individuals through 424 winter den seasons. We examined 183 litters consisting of 455 cubs. Primiparity occurred at mean and modal ages of 3.8 and 3 years, respectively. Composite mean litter size was 2.49 (SE = 0.06) cubs/litter; 3‐ and 4‐year‐olds had smaller litters than older bears. We tracked reproductive synchrony using 5 indices and documented a resetting of this synchrony, likely in response to hard‐mast failure. The amplitude of oscillations in synchrony indices dampened through time after the synchronizing events. Documentation and quantification of relationships between nutrient availability, reproduction and population dynamics can be used to inform population modeling efforts and more accurately forecast harvest. © 2011 The Wildlife Society.

The skeleton gets a (reproductive) life

The skeleton gets a (reproductive) life

Evaluating leaf litter beetle data sampled by Winkler extraction from Atlantic forest sites in southern Brazil

Evaluating leaf litter beetle data sampled by Winkler extraction from Atlantic forest sites in southern Brazil. To evaluate the reliability of data obtained by Winkler extraction in Atlantic forest sites in southern Brazil, we studied litter beetle assemblages in secondary forests (5 to 55 years after abandonment) and old-growth forests at two seasonally different points in time. For all regeneration stages, species density and abundance were lower in April compared to August; but, assemblage compo- sition of the corresponding forest stages was similar in both months. We suggest that sampling of small litter inhabiting beetles at different points in time using the Winkler technique reveals identical ecological patterns, which are more likely to be influenced by sample incompleteness than by differences in their assemblage composition. A strong relationship between litter quantity and beetle occurrences indicates the importance of this variable for the temporal species density pattern. Additionally, the sampled beetle material was compared with beetle data obtained with pitfall traps in one old-growth forest. Over 60% of the focal species captured with pitfall traps were also sampled by Winkler extraction in different forest stages. Few beetles with a body size too large to be sampled by Winkler extraction were only sampled with pitfall traps. This indicates that the local litter beetle fauna is domi- nated by small species. Hence, being aware of the exclusion of large beetles and beetle species occurring during the wet season, the Winkler method reveals a reliable picture of the local leaf litter beetle community.

Litter Size Variation in Hypothalamic Gene Expression Determines Adult Metabolic Phenotype in Brandt's Voles (Lasiopodomys brandtii)

BackgroundEarly postnatal environments may have long-term and potentially irreversible consequences on hypothalamic neurons involved in energy homeostasis. Litter size is an important life history trait and negatively correlated with milk intake in small mammals, and thus has been regarded as a naturally varying feature of the early developmental environment. Here we investigated the long-term effects of litter size on metabolic phenotype and hypothalamic neuropeptide mRNA expression involved in the regulation of energy homeostasis, using the offspring reared from large (10–12) and small (3–4) litter sizes, of Brandt's voles (Lasiopodomys brandtii), a rodent species from Inner Mongolia grassland in China.Methodology/Principal FindingsHypothalamic leptin signaling and neuropeptides were measured by Real-Time PCR. We showed that offspring reared from small litters were heavier at weaning and also in adulthood than offspring from large litters, accompanied by increased food intake during development. There were no significant differences in serum leptin levels or leptin receptor (OB-Rb) mRNA in the hypothalamus at weaning or in adulthood, however, hypothalamic suppressor of cytokine signaling 3 (SOCS3) mRNA in adulthood increased in small litters compared to that in large litters. As a result, the agouti-related peptide (AgRP) mRNA increased in the offspring from small litters.Conclusions/SignificanceThese findings support our hypothesis that natural litter size has a permanent effect on offspring metabolic phenotype and hypothalamic neuropeptide expression, and suggest central leptin resistance and the resultant increase in AgRP expression may be a fundamental mechanism underlying hyperphagia and the increased risk of overweight in pups of small litters. Thus, we conclude that litter size may be an important and central determinant of metabolic fitness in adulthood.

Higher White Adipocyte Area and Lower Leptin Production in Adult Rats Overfed During Lactation

Litter size reduction during lactation is a good model for childhood obesity since it induces overnutrition and programming for obesity at adulthood. Adult offspring develop higher fat mass content, hyperinsulinemia and insulin resistance, hypertension, lower HDL cholesterol, hyperphagia, and leptin resistance. Leptin resistance is often associated with hyperleptinemia. Although we observed higher SOCS3 and lower STAT3 in the hypothalamus of rats raised in small litters featuring a central leptin resistance, they showed unexpected normoleptinemia at 180 days old. Then, to clarify why early overfed rats did not develop hyperleptinemia when adult, we studied the leptin production by the visceral and subcutaneous adipose tissue and skeletal muscle as well as the morphology in the 2 different fat depots. To induce EO, litter size was reduced to 3 pups/litter (SL group) on the 3 (rd) day of life. In controls (NL group), litter size was adjusted to 10 pups/litter. Rats were killed at 180 days old. The programming of adipose tissue morphology by early overnutrition is specific between the different fat depots with hypertrophy only in the visceral compartment. In addition, the visceral adipocyte showed lower leptin content that may indicate a reduced leptin synthesis. These data suggest that adipocytes from SL rats are dysfunctional, since a higher leptin production in larger adipose cells is expected. In conclusion, postnatal nutrition is determinant for future leptin production by different fat depots as well as adipocyte morphology. These changes seem to be related to the severity of obesity and its metabolic consequences.