Early puberty in short-haired Guinea pigs kept in laboratory animal facilities

Lab animals, such as Guinea pigs (Cavia porcellus), are crucial for scientific development, as they play an important role in the development and quality control chain of vaccines and drugs distributed by the Brazilian public health system. Investigating their biological and physiological parameters is fundamental to raise and keep these animals, so the handling of the facilities that hold them can be updated whenever new information comes up, with the well-being of the animals and alignment with the 3 Rs in mind. In the search for understanding reproductive aspects of Guinea pigs, the present study had the main goal of studying puberty by means of estrous cycle analysis in short-haired Guinea pigs. Guinea pigs have a vaginal occlusive membrane that covers the vaginal orifice. Its rupture takes place gradually and naturally, moments before labor and during estrus. The present study followed 42 females as for the presentation of the vaginal occlusive membrane. Once the membranes ruptured spontaneously, a swab was collected to study vaginal cytology. Membrane rupture was observed in 39 females; six females showed membrane rupture with less than 21 days of age (17 to 21 days). Twenty-three females were characterized as being in estrus due to cytology showing a prevalence of anucleated superficial cells. One of these females was younger than 21 days old. The opening of the vaginal occlusive membrane took place most frequently in intervals between 17 and 18 days, and the membrane remained open between one and three consecutive days. It was possible to follow three cycles of membrane opening on six females. The present study showed the need to adapt handling guidelines for C. porcellus kept in research animal facilities. The early age of puberty imposes the need of separate the female daughters from their fathers at 16 days old.     

Metabolic and reproductive characteristics of replacement beef heifers subjected to an early-weaning regimen involving high-concentrate feeding

In an effort to better understand the consequences of early weaning (EW) for replacement beef heifers, a two-phase experiment was conducted investigating the impact on metabolic function and documenting reproductive characteristics. In phase 1, Angus×Simmental heifers (n=35) were stratified by BW and sire, and randomly assigned to either a normal weaning (NW, n=18) or EW (n=17) treatment. EW heifers were weaned at 107±3 days of age and provided access to a concentrate-based ration ad libitum with limit-fed mixed grass hay. NW heifers remained with their dams until 232±3 days of age, at which point heifers from both treatments were comingled and grazed on mixed summer pasture. Following NW, weekly blood samples were collected from all heifers for progesterone analyses used to determine the onset of puberty. Pelvic and ovarian size was measured before breeding. All heifers were subjected to an estrous synchronization protocol with timed artificial insemination (AI) at 437±4 days of age. During phase 2 of the experiment, a subset of pregnant heifers (n=16) were divided into two replicates and subjected to a glucose tolerance test, epinephrine challenge and progesterone clearance analysis. Neither age nor BW at puberty differed between EW and NW heifers. Likewise, no differences in pelvic area or ovarian size were observed. Thus, it appears that the reproductive maturity of EW and NW heifers was similar. Heifers studied during phase 2 of the experiment were restricted to those that had become pregnant to their first AI. Within this cohort, EW heifers tended to have lower overall circulating progesterone concentrations than those that were NW (P=0.14). Aspects of glucose and insulin dynamics were also altered, as EW heifers tended to have lower baseline glucose concentrations (P=0.10) despite similar baseline insulin concentrations. Compared with NW heifers, EW heifers had lower insulin area under the curve (P<0.05), which was partly the result of a tendency for lower peak insulin concentrations (P=0.11). Results of the glucose tolerance test indicate that a lesser insulin response was necessary to properly clear the glucose in the EW heifers, suggesting enhanced insulin sensitivity. Collectively, these results indicate that EW is not detrimental for the growth or reproductive development of replacement beef heifers, although some differences in glucose and insulin dynamics persist into adulthood.     

Genome‐scan analysis for genetic mapping of quantitative trait loci underlying birth weight and onset of puberty in doe kids (Capra hircus)

The objective of this study was to locate quantitative trait loci (QTL) causing variation in birth weight and age of puberty of doe kids in a population of Rayini cashmere goats. Four hundred and thirty kids from five half‐sib families were genotyped for 116 microsatellite markers located on the caprine autosomes. The traits recorded were birth weight of the male and female kids, body weight at puberty, average daily gain from birth to age of puberty and age at puberty of the doe kids. QTL analysis was conducted using the least squares interval mapping approach. Linkage analysis indicated significant QTL for birth weight on Capra hircus chromosomes (CHI) 4, 5, 6, 18 and 21. Five QTL located on CHI 5, 14 and 29 were associated with age at puberty. Across‐family analysis revealed evidence for overlapping QTL affecting birth weight (78 cM), body weight at puberty (72 cM), average daily gain from birth to age of puberty (72 cM) and age at puberty (76 cM) on CHI 5 and overlapping QTL controlling body weight at puberty and age at puberty on CHI 14 at 18–19 cM. The proportion of the phenotypic variance explained by the detected QTL ranged between 7.9% and 14.4%. Confirming some of the previously reported results for birth weight and growth QTL in goats, this study identified more QTL for these traits and is the first report of QTL for onset of puberty in doe kids.     

Impaired reproduction in adult male, but not female, rats following juvenile treatment with the aromatase inhibitor, exemestane

BACKGROUND: Exemestane is an irreversible steroidal inhibitor of cytochrome‐P450 aromatase required for estrogen synthesis. The safety of the drug in the pediatric population, particularly in males, has not previously been evaluated. Given the increased interest in treating children with aromatase inhibitors, we undertook a study in rats to assess the potential for exemestane to alter reproductive development and function when administered to juveniles. METHODS: Male and female rats were treated with exemestane at doses anticipated to produce exposures approximately 2‐ and 35‐fold the expected clinical plasma exposure in young adult males during the period of reproductive maturation. After maturation, treated rats were mated to evaluate the potential impact on reproductive function. RESULTS AND CONCLUSION: There were no effects on sexual maturation in either sex or on female reproductive function. Treatment of juvenile male rats caused increased cohabitation time and decreased copulation rates; pregnancy rates and litter size were not affected in rats that mated. Decreased testis (10–15%) and epididymis (20–30%) weights, and decreased Sertoli cell numbers were noted at all doses. This indicates that exemestane can reduce Sertoli cell proliferation during maturation. The sensitive window for this effect is expected to be limited to the period of Sertoli cell proliferation, which is completed by around postnatal day 15 in rats and before puberty in humans. Treatment beginning at a later time relative to the window for Sertoli cell proliferation or for a longer duration is not expected to have additional adverse effect as the effect was not shown to be degenerative. Birth Defects Res (Part B) 92:304–313, 2011. © 2011 Wiley‐Liss, Inc.     

Adolescent brain maturation is necessary for adult‐typical mesocorticolimbic responses to a rewarding social cue

The interpretation of social cues must change during adolescence in order to promote appropriate social interactions in adulthood. For example, adult, but not juvenile, male Syrian hamsters find female pheromones contained in vaginal sections (VS) rewarding, and only adult hamsters engage in sexual behavior with a receptive female. We previously demonstrated that the rewarding value of VS is both testosterone‐ and dopamine‐dependent. Additionally, VS induces Fos expression throughout the mesocorticolimbic circuit in adult but not juvenile hamsters. In this study, we determined whether or not treatment of juvenile male hamsters with testosterone is sufficient to promote adult‐like neural responses to VS. Juvenile and adult male hamsters were gonadectomized and given empty or testosterone‐filled subcutaneous capsules for 1 week. Hamsters were then exposed to either clean or VS‐containing mineral oil on their nares, and brains were collected 1 h later for immunohistochemistry to visualize Fos and tyrosine hydroxylase immunoreactive cells. Testosterone treatment failed to promote adult‐typical patterns of Fos expression in juvenile hamsters; indeed, in some brain regions, juveniles exposed to VS expressed less Fos compared to age‐matched controls while, as expected, adults exposed to VS expressed greater Fos compared to age‐matched controls. Age‐related changes in tyrosine hydroxylase expression were also observed. These data indicate that testosterone cannot activate the adult‐typical pattern of Fos expression in response to female social cues in prepubertal males, and that additional neural maturation during adolescence is required for adult‐typical mesocorticolimbic responses to female pheromones. © 2013 Wiley Periodicals, Inc. Develop Neurobiol 73:856–869, 2013     

Glycerol Dehydrogenase from Cellulomonas sp. NT3060: Purification and Characterization

NAD⁺-dependent glycerol dehydrogenase from Cellulomonas sp. NT3060 was purified by a procedure of 10 steps involving crystallization. Dihydroxyacetone was identified as the oxidation product of glycerol with the enzyme. The purified enzyme did not lose activity on heating below 60°C. The enzyme oxidized other alcohols such as 1,2-propanediol, 2,3-butanediol and glycerol-α-monochlorohydrin, beside glycerol. The enzyme activity was inhibited by p-chloromercuribenzoate, Zn²⁺, Cu²⁺ and Cd²⁺. Oxidation of glyberol was activated by Na⁺ and reduction of dihydroxyacetone was activated by K⁺ at pH 7.5.     

Isolation of Oligosaccharides Corresponding to the Branching-point of Konjac Mannan

Ultracentrifugically homogeneous glucomannan acetate derived from konjac mannan was subjected to acetolysis. Besides β-1,4-linked oligosaccharides composed of D-mannose and/or D-glucose, three oligosaccharides corresponding to the branching point of the polysaccharide were isolated and identified as (1) 3-O-β-D-mannopyranosyl-D-mannose, (2) O-β-D-mannopyranosyl-(1→4)-O-β-D-mannopyranosyl-(1→3)-D-mannose, and (3) O-β-D- mannopyranosyl-(1→3)-O-β-D-mannopyranosyl-(1→4)-D-glucose. The average chain length (CL) was, moreover, determined to be about 46 by methylation analysis. The structural pattern of the glucomannan, including the branching point, is discussed.     

Effects of dietary menhaden oil on growth and reproduction in gilts farrowed by sows that consumed diets containing menhaden oil during gestation and lactation

In sows, n-3 fatty acids increase litter sizes, however, effects on gilt reproductive development have not been adequately studied. Moreover, not determined are effects of feeding n-3 fatty acids to sows on reproduction in offspring. The objective here was to determine effects of 4% dietary menhaden oil on growth and puberty in gilts farrowed by sows fed menhaden oil. Sows (n = 44) were assigned to: (1) control gestation and lactation diets, or (2) diets including menhaden oil. For primiparous sows only, total litter size and born alive were greater (P < 0.05) in females fed menhaden oil. Conversely, pigs from primiparous controls were heavier (P < 0.05) than pigs from primiparous sows fed menhaden oil (parity by diet interactions, P < 0.01). Diet did not affect (P > 0.20) other sow and litter characteristics. At weaning, 84 gilts from control- or menhaden oil sows were placed three gilts per pen and provided control diets or diets containing menhaden oil. Nursery and grow-finish feed intake and feed efficiency were similar (P > 0.21) for gilts from the different sows and weight gain was similar (P > 0.24) for gilts fed control or menhaden diets. Gilts fed menhaden oil tended to eat less in the nursery (1.18±0.08 kg v. 0.98±0.08 kg; P = 0.09) and overall (1.83±0.04 kg v. 1.72±0.04 kg; P = 0.06). Thus, overall feed to gain was greater (2.52±0.03 v. 2.33±0.03; P < 0.01) and nursery (2.12±0.04 v. 1.80±0.04; P = 0.10) and grow-finish (3.07±0.19 v. 2.58±0.19; P = 0.08) feed to gain tended to be greater, for control gilts. Age at puberty was greater (P = 0.02) for gilts from menhaden oil-fed sows (205.1±3.2 days) compared to gilts from controls (193.9±3.2 days) and tended to be greater (P = 0.09), for controls (203.5±3.2 days) compared to gilts fed menhaden oil (195.5±3.2 days). A tendency existed (P = 0.09) for greater follicular fluid in gilts fed menhaden oil, however, ovulation rate and ovarian, luteal and uterine weights were not affected by sow diet, gilt diet or the interaction (P > 0.23). Feeding gilts menhaden oil enhanced feed efficiency and hastened puberty onset. Gilts from sows consuming menhaden oil exhibited delayed puberty and retaining females from sows fed this feedstuff may be ill advised.     

The integrity of sperm chromatin in young tropical composite bulls

Sperm chromatin fragmentation is associated with subfertility, but its relationship with age progression in young bulls is poorly understood. The objective was to assess sperm chromatin fragmentation during the early post-pubertal development of 20 tropical composite bulls, using a sperm chromatin structure assay (SCSA) and sperm-bos-halomax (SBH). Bulls were subjected to bull breeding soundness evaluation (BBSE) at mean ages of 13, 18, and 24 mo. Traits measured included liveweight (WT), body condition score (BCS) and scrotal circumference (SC). Semen samples were collected by electroejaculation and assessed for mass activity (MA), motility (Mot), concentration (conc), sperm morphology and chromatin fragmentation. Concentration (r = 0.34, P = 0.0076), Mot (r = 0.36, P = 0.0041) and percentage of morphologic normal sperm (percent normal sperm (PNS); r = 0.31, P = 0.0132) were positively correlated with age. The percentage of sperm with proximal droplets (PD) was negatively correlated with age (r = −0.28, P = 0.0348), whereas neither SCSA nor SBH results were significantly correlated with age. The percentage of sperm with chromatin fragmentation using SCSA was correlated with PNS (r = −0.53, P < 0.0001), the percentage of sperm with head abnormalities (r = 0.68, P < 0.0001) and the percentage of intact sperm (Int) with SBH (r = −0.26, P = 0.0456). In summary, for assessment of sperm chromatin fragmentation, samples could be equally collected at 13, 18 or 24 mo of age, as results did not vary with age.