Differences in hormone localisation patterns of K and L type enteroendocrine cells in the mouse and pig small intestine and colon

 This study has investigated the patterns of colocalisation of the conventional K cell marker, glucagon-like insulinotropic peptide (GIP), and the L cell markers, glucagon like peptide-1 (GLP-1) and peptide YY (PYY), in enteroendocrine cells (EEC) of the small intestine and colon of mouse and pig. All combinations of the hormones, 3 in a cell, 2 in a cell and 1 at a time, were encountered. In both species, the three most common EEC types contained (1) both GLP-1 and PYY but not GIP, (2) GLP-1 alone or (3) GIP plus GLP-1 without PYY. Few GIP plus PYY cells and rare cells containing all 3 hormones were encountered. Gradients of cell types occurred along the intestine. For example, in mouse, there were no PYY cells in the duodenum and few in the jejunum, but >50 % of labelled EEC in the distal ileum and colon were PYY immunoreactive. By contrast, over 40 % of EEC in the pig duodenum contained PYY, and most also contained either GLP-1 or GIP. The gradient in pig was less pronounced. It is concluded that the traditional classification of K and L cells requires revision, and that there are major inter-species differences in the patterns of colocalisation of hormones that have been used to characterise K and L cells.

Effects of stress on reproduction in non-rodent mammals : the role of glucocorticoids and sex differences

The means by which stress influences reproduction is not clearly understood, but may involve a number of endocrine, paracrine and neural systems. Stress impacts on the reproductive axis at the hypothalamus (to affect GnRH secretion) and the pituitary gland (to affect gonadotrophin secretion), with direct effects on the gonads being of less importance. Different stressors have different effects and there are differences in response to short- and long-term stress. Many short-term stresses fail to affect reproduction and there are reports of stimulatory effects of some 'stressors'. There are species differences in the way that specific stressors affect reproduction. Sex differences in the effects of a particular stressor have been delineated and these may relate to effects of stress at different levels of the hypothalamo-pituitary axis. The significance of stress-induced secretion of cortisol varies with species. In some instances, there appears to be little impact of short-term increases in cortisol concentrations and protracted increases in plasma concentration seem to be required before any deleterious effect on reproduction is apparent. Issues of sex, sex steroid status, type of stressor and duration of stress need to be considered to improve understanding of this issue.

Histological assessment of intermediate - and long-term creatine monohydrate supplementation in mice and rats

Creatine monohydrate (CrM) supplementation appears to be relatively safe based on data from short-term and intermediate-term human studies and results from several therapeutic trials. The purpose of the current study was to characterize pathological changes after intermediate-term and long-term CrM supplementation in mice [healthy control and SOD1 (G93A) transgenic] and rats (prednisolone and nonprednisolone treated). Histological assessment (18-20 organs/tissues) was performed on G93A mice after 159 days, and in Sprague-Dawley rats after 365 days, of CrM supplementation (2% wt/wt) compared with control feed. Liver histology was also evaluated in CD-1 mice after 300 days of low-dose CrM supplementation (0.025 and 0.05 g · kg^-1 · day^-1) and in Sprague-Dawley rats after 52 days of CrM supplementation (2% wt/wt) with and without prednisolone. Areas of hepatitis were observed in the livers of the CrM-supplemented G93A mice (P < 0.05), with no significant inflammatory lesions in any of the other 18-20 tissues/organs that were evaluated. The CD-1 mice also showed significant hepatic inflammatory lesions (P < 0.05), yet there was no negative effect of CrM on liver histology in the Sprague-Dawley rats after intermediate-term or long-term supplementation nor was inflammation seen in any other tissues/organs (P = not significant). Dietary CrM supplementation can induce inflammatory changes in the liver of mice, but not rats. The observed inflammatory changes in the murine liver must be considered in the evaluation of hepatic metabolism in CrM-supplemented mice. Species differences must be considered in the evaluation of toxicological and physiological studies.

Enhanced proliferation of human skeletal muscle precursor cells derived from elderly donors cultured in estimated physiological (5%) oxygen

Human skeletal muscle precursor cells (myoblasts) have significant therapeutic potential and are a valuable research tool to study muscle cell biology. Oxygen is a critical factor in the successful culture of myoblasts with low (1–6%) oxygen culture conditions enhancing the proliferation, differentiation, and/or viability of mouse, rat, and bovine myoblasts. The specific effects of low oxygen depend on the myoblast source and oxygen concentration; however, variable oxygen conditions have not been tested in the culture of human myoblasts. In this study, muscle precursor cells were isolated from vastus lateralis muscle biopsies and myoblast cultures were established in 5% oxygen, before being divided into physiological (5%) or standard (20%) oxygen conditions for experimental analysis. Five percent oxygen increased proliferating myoblast numbers, and since low oxygen had no significant effect on myoblast viability, this increase in cell number was attributed to enhanced proliferation. The proportion of cells in the S (DNA synthesis) phase of the cell cycle was increased by 50%, and p21^Cip1 gene and protein expression was decreased in 5 versus 20% oxygen. Unlike in rodent and bovine myoblasts, the increase in myoD, myogenin, creatine kinase, and myosin heavy chain IIa gene expression during differentiation was similar in 5 and 20% oxygen; as was myotube hypertrophy. These data indicate for the first time that low oxygen culture conditions stimulate proliferation, whilst maintaining (but not enhancing) the viability and the differentiation potential of human primary myoblasts and should be considered as optimum conditions for exvivo expansion of these cells.

Evolution of cuticular hydrocarbon diversity in ants

The cuticular hydrocarbons (CHCs) of ants provide important cues for nestmate and caste recognition. There is enormous diversity in the composition of these CHCs, but the manner in which this diversity has evolved is poorly understood. We gathered data on CHC profiles for 56 ant species, relating this information to their phylogeny. We deduced the mode of evolution of CHC profiles by reconstructing character evolution and then relating the number of changes in CHC components along each branch of the phylogeny to the length of the branch. There was a strong correlation between branch length and number of component changes, with fewer changes occurring on short branches. Our analysis thereby indicated a gradual mode of evolution. Different ant species tend to use specific CHC structural types that are exclusive of other structural types, indicating that species differences may be generated in part by switching particular biosynthetic pathways on or off in different lineages. We found limited, and contradictory, evidence for abiotic factors (temperature and rainfall) driving change in CHC profiles.

Optimizing survey effort for burrow-nesting seabirds

Regular monitoring of seabird populations is necessary to improve our understanding of their responses to environmental change and inform conservation management. However, given the difficulty in accessing remote breeding sites and the limited resources typically available to land managers, conducting regular, extensive surveys of seabird populations is often not feasible. Our objective was to determine the minimum survey effort required to obtain accurate and precise population estimates of Short-tailed Shearwaters (Ardenna tenuirostris) and Little Penguins (Eudyptula minor), two abundant burrowing seabird species in southeastern Australia, by comparing bootstrapped means and confidence intervals under different sampling regimes on four islands. We found that, in many cases, survey effort (the proportion of transects and quadrats along transects surveyed) could be reduced. For Short-tailed Shearwaters, reducing the number of transects resulted in a maximum difference of 15% between the means at full survey effort and two levels of reduced survey effort. Means differed by <3% when we halved the number of quadrats. For Little Penguins, reducing the number of transects and quadrats by 50% resulted in differences of 7-40% and 4-34%, respectively, between the full and reduced survey effort means. Confidence intervals generally increased with decreasing survey effort for both species. Differences in required survey effort between the two species in our study may have been due to differences in burrow distribution on islands, with Short-tailed Shearwater burrows generally uniformly distributed on each island and Little Penguin burrows typically occurring in patches. These would be influenced by island-specific characteristics in concert with habitat preferences, population size, and seasonal variation in seabird abundance. Stratified sampling did not increase survey accuracy and simulations showed that large reductions in survey effort could be made under a pseudo-random sampling regime, with mean abundance estimates similar at most levels of survey effort. For both species, reducing the proportion of pseudo-random quadrats to 50% and 25% of the full survey effort produced confidence intervals of 12% and 21%, respectively, of the maximum, whereas a survey effort of 10% produced confidence intervals of up to 36% of the maximum for both species. A pseudo-random sampling regime would maximize survey efficiency because considerably fewer quadrats would be required and allow development of more efficient sampling protocols and regimes.

Gastrointestinal microbiota of seabirds

This work provided novel insight into the previously uncharacterised microbial composition of Antarctic and temperate penguins and procellariiform seabirds. Using real time PCR and 16S pyrosequencing the results identified significant inter- and intra-species differences, insight into the successional changes that occur during development and how fasting influences microbial composition

Flipper strokes can predict energy expenditure and locomotion costs in free-ranging northern and Antarctic fur seals

Flipper strokes have been proposed as proxies to estimate the energy expended by marine vertebrates while foraging at sea, but this has never been validated on free-ranging otariids (fur seals and sea lions). Our goal was to investigate how well flipper strokes correlate with energy expenditure in 33 foraging northern and Antarctic fur seals equipped with accelerometers, GPS, and time-depth recorders. We concomitantly measured field metabolic rates with the doubly-labelled water method and derived activity-specific energy expenditures using fine-scale time-activity budgets for each seal. Flipper strokes were detected while diving or surface transiting using dynamic acceleration. Despite some inter-species differences in flipper stroke dynamics or frequencies, both species of fur seals spent 3.79 ± 0.39 J/kg per stroke and had a cost of transport of ~1.6-1.9 J/kg/m while diving. Also, flipper stroke counts were good predictors of energy spent while diving (R(2) = 0.76) and to a lesser extent while transiting (R(2) = 0.63). However, flipper stroke count was a poor predictor overall of total energy spent during a full foraging trip (R(2) = 0.50). Amplitude of flipper strokes (i.e., acceleration amplitude × number of strokes) predicted total energy expenditure (R(2) = 0.63) better than flipper stroke counts, but was not as accurate as other acceleration-based proxies, i.e. Overall Dynamic Body Acceleration.

Differences in foraging strategy and maternal behaviour between two sympatric fur seal species at the Crozet Islands

Marine top-predators such as marine mammals forage in a heterogeneous environment according to their energetic requirements and to the variation in environmental characteristics. In this study, the behaviour of breeding females in 2 sympatric fur seal species, Antarctic fur seal Arctocephalus gazella and Subantarctic fur seal A. tropicalis, was investigated in relation to foraging effort. Foraging effort was hypothesised to be greater in Antarctic fur seal than in Subantarctic fur seal due to their shorter lactation period. Using satellite telemetry, time-depth recorders and satellite images of sea-surface temperature and chlorophyll a concentration, the foraging grounds, the at-sea activity budgets and the environmental features were determined for both species breeding on the Crozet Archipelago. Foraging cycle duration was similar for the 2 species, and the seals exhibited similar at-sea activity budgets. Only the proportion of time spent at sea was higher in Antarctic fur seals. Separate foraging areas were identified for the 2 species. Antarctic fur seal distribution was related to bathymetric features, while we did not find any direct relationship between chlorophyll a concentration and seal foraging areas. Our results suggest that Antarctic fur seals tend to respond to the higher needs of their pups by having a higher foraging efficiency and concentrating their foraging activity in the most productive areas.

Stress and reproduction: central mechanisms and sex differences in non-rodent species

Despite extensive research, the mechanisms by which stress affects reproduction are unknown. Activation of stress systems could potentially influence reproduction at any level of the hypothalamo-pituitary gonadal axis. Nonetheless, the predominant impact is on the secretion of gonadotrophin releasing hormone (GnRH) from the brain and the secretion of the gonadotrophins, luteinizing hormone (LH) and follicle stimulating hormone (FSH), from the gonadotrophs of the anterior pituitary gland. When stress is prolonged, it is likely that secretion of the gonadotrophins will be suppressed but the effects of acute stress or repeated acute stress are not clear. Different stressors activate different pathways for varying durations, and the actions of stress vary with sex and are influenced by the predominance of particular sex steroids in the circulation. The mechanisms by which stress influences reproduction are likely to involve complex interactions between a number of central and peripheral pathways and may be different in males and females. To understand these mechanisms, it is important to determine the stress pathways that are activated by particular stressors and to establish how these pathways affect the secretion and actions of GnRH. Furthermore, there is a need to know how stress influences the feedback actions of gonadal steroids and inhibin.

Previous land use and climate influence differences in soil organic carbon following reforestation of agricultural land with mixed-species plantings

Reforestation of agricultural land with mixed-species environmental plantings (native trees and shrubs) can contribute to mitigation of climate change through sequestration of carbon. Although soil carbon sequestration following reforestation has been investigated at site- and regional-scales, there are few studies across regions where the impact of a broad range of site conditions and management practices can be assessed. We collated new and existing data on soil organic carbon (SOC, 0-30 cm depth, N = 117 sites) and litter (N = 106 sites) under mixed-species plantings and an agricultural pair or baseline across southern and eastern Australia. Sites covered a range of previous land uses, initial SOC stocks, climatic conditions and management types. Differences in total SOC stocks following reforestation were significant at 52% of sites, with a mean rate of increase of 0.57 ± 0.06 Mg C ha-1 y-1. Increases were largely in the particulate fraction, which increased significantly at 46% of sites compared with increases at 27% of sites for the humus fraction. Although relative increase was highest in the particulate fraction, the humus fraction was the largest proportion of total SOC and so absolute differences in both fractions were similar. Accumulation rates of carbon in litter were 0.39 ± 0.02 Mg C ha-1 y-1, increasing the total (soil + litter) annual rate of carbon sequestration by 68%. Previously-cropped sites accumulated more SOC than previously-grazed sites. The explained variance differed widely among empirical models of differences in SOC stocks following reforestation according to SOC fraction and depth for previously-grazed (R2 = 0.18-0.51) and previously-cropped (R2 = 0.14-0.60) sites. For previously-grazed sites, differences in SOC following reforestation were negatively related to total SOC in the pasture. By comparison, for previously-cropped sites, differences in SOC were positively related to mean annual rainfall. This improved broad-scale understanding of the magnitude and predictors of changes in stocks of soil and litter C following reforestation is valuable for the development of policy on carbon markets and the establishment of future mixed-species environmental plantings.

Molecular taxonomy and phylogenetics of some species of Australian palaemonid shrimps

The evolutionary history and classification of the palaemonid shrimps has been the subject of constant speculation and debate. At present, all major systematic treatments have been based on morphological characteristics. To help resolve the phylogenetic relationships, and thus enable the creation of a classification system that reflects evolutionary history, a region of the 16S mitochondrial rRNA gene was sequenced for a number of Australian Palaemonidae. The resulting phylogenetic analyses indicated the presence of major anomalies in the current classification of Australian Palaemonidae. Significantly, three species belonging to three separate genera, Macrobrachium intermedium, Palaemon serenus, and Palaemonetes australis, are closely related, with genetic differences more characteristic with that of congeneric species. The results also demonstrate non-monophyly in Australian palaemonids with respect to both Palaemonetes and Macrobrachium.

Long-term flowering patterns of melliferous Eucalyptus (Myrtaceae) species

The flowering patterns of 28 Victorian melliferous (honey-producing) eucalypts were investigated by using long-term observations of highly experienced, commercial apiarists. Frequency, timing, duration and intensity of flowering were determined, as were spatial differences within and among species. Data were obtained by face-to-face interviews with 25 Victorian apiarists, each of whom had operated a minimum of 350 hives for a minimum of 30 years. Flowering frequency ranged from 1 to 7 years, and most species flowered once every 2–4 years. Long-term flowering frequency, timing and duration were reported as constant, although short-term perturbations could occur. Most melliferous species flowered during spring and summer for a period of 3 months or more. Only few species had shorter flowering periods. Information provided by apiarists compared well with available published information (e.g. flowering period reported in field guides) and revealed a reliable, largely untapped source of long-term data, the use of which could benefit many ecological research endeavours.

Mitochondrial DNA diversity of broodstock of two indigenous mahseer species, Tor tambroides and T. douronensis (Cyprinidae) cultured in Sarawak, Malaysia

Tor tambroides and T. douronensis, locally referred to as empurau and semah, respectively, are high valued mahseer species, indigenous to Sarawak, East Malaysia, with an aquaculture potential and of conservational value. Direct sequencing of mitochondrial DNA (mtDNA) 16S rRNA gene region (542 bp) was used to investigate genetic variation of T. tambroides and T. douronensis broodstock collected from different geographic locations in Sarawak and maintained at the Indigenous Fish Research and Production Center (IFRPC), Tarat, Sarawak, Malaysia. A total of 11 unique haplotypes were identified, of which six were detected in T. tambroides, and five in T. douronensis. Overall, nucleotide diversity (π) was low, ranging from 0.000 to 0.006, and haplotype diversity (h) ranged from 0.000 to 0.599. Although the analysis failed to detect genetic variation amongst populations of T. tambroides (significant pairwise FST was found for only one test, but pairwise haplotype frequencies were not statistically significant), substantial inter-population divergence among T. douronensis was recognised, especially those originating from different river systems (pairwise F_ST = 0.754 to 1.000, P < 0.05). Fixed haplotype differences were found in one population of T. douronensis. Average nucleotide divergence between T. tambroides and T. douronensis was 0.018, similar to the amount recognised between T. tambroides and the outgroup T. khudree (0.017). In addition, phylogenetic analysis revealed that the T. douronensis mtDNA consisted of two highly divergent clusters (0.020), one of which is more closely related to T. tambroides rather than with the other group of haplotypes of the conspecifics. The findings from the present study have important implications for aquaculture, management and conservation of these two species. The data also raise some concerns regarding the taxonomic status of T. douronensis, which needs to be addressed.