Search

Remote camera-traps are commonly used to estimate the abundance, diversity, behavior and habitat use of wildlife in an inexpensive and nonintrusive manner. Because of the increasing use of remote-cameras in wildlife studies, students interested in wildlife biology should be exposed to the use of remote-cameras early in their academic careers. Although there is a rich literature on the use of remote-cameras in wildlife studies, few have provided meaningful examples within an academic course setting. Due to the time constraint of a typical semester, many laboratory exercises generate data sufficient for the activity but lack inference to actual wildlife populations. This article describes a series of laboratory exercises that are both useful to student learning and provide relevant biological data. Students use remote-cameras to measure diversity, diel behavior (i.e. over a 24-hour period) and the relative abundance of mammals in a biological corridor. Other abundance methods such as mark-recapture or random encounter models that require marked individuals and/or extensive temporal and spatial methodology are often not practical in a course framework. The approach described in this article teaches students about research design and local wildlife abundance and behavior, using simple methodologies employed over a three-lab period.

This paper describes a series of laboratory exercises for upper level biology courses, independent research and/or honors programs. Students sampled fish from a local water body with the assistance of a local fish and wildlife agency. Tissue samples from collected fish were utilized to obtain estimates of the stable isotopes delta[superscript 13]C and delta[superscript 15]N. An open-source mass balance model was utilized to estimate fish diets from isotope values. Isotopes were also used to measure trophic position (TP) and evaluate dietary niche overlap between native and introduced species. It was found that delta[superscript 15]N concentrations increased with each trophic level starting with benthic algae (lowest) to omnivores, then primary carnivores and ultimately piscivores (highest). Estimates of delta[superscript 13]C suggested all collected taxa utilized littoral habitats for feeding purposes. Native chain pickerel ("Esox niger") had the highest delta[superscript 15]N and TP estimates at 14.4 and 4.11, respectively. Nonnative largemouth bass ("Micropterus salmoides") delta[superscript 15]N and TP estimates were 13.56 and 3.86, respectively. Model estimates suggested that chain pickerel and largemouth bass diets were partitioned: pickerel consumed only fish and bass consumed principally invertebrates with some intermittent fish contributions. This lab helped integrate multiple disciplines, provide practical experience, and encourage critical-thinking skills while educating students about trophic ecology. (Contains 4 figures and 1 table.)

Study Question: Does the type of incubator used to culture human preimplantation embryos affect development to the blastocyst stage and alter amino acid utilization of embryos in assisted reproduction?Summary Answer: Culturing embryos in a time lapse system (TLS) was associated with a higher Day 5 blastocyst formation rate and altered amino acid utilization when measured from Day 3 to Day 5 compared to the standard benchtop incubator.What Is Known Already: Culture environment is known to be important for the developing preimplantation embryo. TLSs provide a stable milieu allowing embryos to be monitored in situ, whereas embryos cultured in standard benchtop incubators experience environmental fluctuations when removed for morphological assessment.Study Design, Size, Duration: A prospective clinical trial randomizing 585 sibling embryos to either the TLS (289 embryos) or the standard benchtop incubator (296 embryos) over a 23-month period in a UK University Hospital Fertility Clinic.Participants/materials, Setting, Methods: Participants were aged 42 years or under, had an antral follicle count of ≥12 and ≥6 2 pronucleate zygotes. Zygotes were cultured individually in 25 µl of medium. Randomized embryos were graded and selected for transfer or cryopreservation on Day 5. For those embryos produced by women who underwent stimulation with recombinant FSH injections and were triggered with hCG, spent medium was collected on Day 5 for amino acid analysis by high pressure liquid chromatography. Clinical pregnancy was defined as the presence of a foetal heart beat on ultrasound scan at 7 weeks.Main Results and the Role Of Chance: Overall, blastocyst formation rate on Day 5 was significantly higher in embryos cultured in the TLS (55%) compared to the standard incubator (45%; P = 0.013). Similarly, there was an increase in the number of blastocysts suitable for cryopreservation in the TLS (31%) compared to the standard incubator (23%; P = 0.032). There was a significant difference in the utilization of 12 amino acids by blastocysts cultured from Day 3 to Day 5 in the TLS compared to the standard incubator. Embryos cultured in the TLS displayed an increased total amino acid utilization (P < 0.001) and reduced amino acid production (P < 0.001) compared to those in the standard incubator. Irrespective of incubator used, embryos fertilized by ICSI depleted significantly more amino acids from the medium compared to those fertilized by conventional IVF. There was no difference in the mean score of blastocysts transferred, or the clinical pregnancy rate after transfer of embryos from either of the incubators.Limitations, Reasons For Caution: The study was not powered to discern significant effects on clinical outcomes.Wider Implications Of the Findings: The metabolism and development of preimplantation embryos is impacted by the type of incubator used for culture. Further research is required to investigate the long-term implications of these findings.Study Funding/competing Interest(s): NIHR Southampton Biomedical Research Centre Commercial and Enterprise Incubator Fund funded this study. The TLS was provided on loan for the study by Vitrolife. The authors declare no conflict of interests.Trial Registration Number: ISRCTN73037149.Trial Registration Date: 12 January 2012.Date Of First Patient’s Enrolment: 21 January 2012. [ABSTRACT FROM AUTHOR]

Low O2 tension is beneficial for human embryonic stem cell (hESC) maintenance but the mechanism of regulation is unknown. HIF-2α was found to bind directly to predicted hypoxic response elements (HREs) in the proximal promoter of OCT4, NANOG and SOX2 only in hESCs cultured under hypoxia (5% O2). This binding induced an array of histone modifications associated with gene transcription while a heterochromatic state existed at atmospheric O2. Interestingly, an enhanced euchromatic state was found when hESCs were exposed to hypoxia followed by 72 hours reoxygenation. This was sustained by HIF-2α which enhanced stemness by binding to an oct-sox cis-regulatory element in the NANOG promoter. Thus, these data have uncovered a novel role of HIF-2α as a direct regulator of key transcription factors controlling self-renewal in hESCs but also in the induction of epigenetic modifications ensuring a euchromatic conformation which enhances the regenerative potential of these cells. [ABSTRACT FROM AUTHOR]

Background/Aims: Human embryonic stem cells (hESCs) are a potential source of cells for treatment of many degenerative diseases, but in culture have a propensity to spontaneously differentiate, possibly due to suboptimal conditions. Culture at low oxygen tensions improves hESC maintenance and regulates carbohydrate metabolism. Hence, a greater understanding of the nutrient requirements of hESCs will allow production of more appropriate culture media. This study aims to investigate the effect of environmental oxygen tension on the amino acid metabolism of hESCs. Methods: The production or depletion of amino acids by hESCs cultured at 5% or 20% oxygen in the presence or absence of FGF2 was measured by reversephase HPLC. Results: Atmospheric oxygen, or removal of FGF2 from hESCs cultured at 5% oxygen, perturbed the uptake or release of individual amino acids and the total amino acid turnover compared to hESCs cultured at 5% oxygen. In particular, serine uptake was reduced at 20% oxygen and by removal of FGF2. Conclusions: Highly pluripotent hESCs, cultured at 5% oxygen, demonstrate a greater amino acid turnover than hESCs cultured at 20% oxygen, or without FGF2. These data suggest that amino acid turnover could be used as a measure of the self-renewal capacity of hESCs. © 2014 S. Karger AG, Basel [ABSTRACT FROM AUTHOR]

Energy metabolism is intrinsic to cell viability but surprisingly has been little studied in human embryonic stem cells (hESCs). The current study aims to investigate the effect of environmental O2 tension on carbohydrate utilisation of hESCs. Highly pluripotent hESCs cultured at 5% O2 consumed significantly more glucose, less pyruvate and produced more lactate compared to those maintained at 20% O2. Moreover, hESCs cultured at atmospheric O2 levels expressed significantly less OCT4, SOX2 and NANOG than those maintained at 5% O2. To determine whether this difference in metabolism was a reflection of the pluripotent state, hESCs were cultured at 5% O2 in the absence of FGF2 for 16 hours leading to a significant reduction in the expression of SOX2. In addition, these cells consumed less glucose and produced significantly less lactate compared to those cultured in the presence of FGF2. hESCs maintained at 5% O2 were found to consume significantly less O2 than those cultured in the absence of FGF2, or at 20% O2. GLUT1 expression correlated with glucose consumption and using siRNA and chromatin immunoprecipitation was found to be directly regulated by hypoxia inducible factor (HIF)-2α at 5% O2. In conclusion, highly pluripotent cells associated with hypoxic culture consume low levels of O2, high levels of glucose and produce large amounts of lactate, while at atmospheric conditions glucose consumption and lactate production are reduced and there is an increase in oxidative metabolism. These data suggest that environmental O2 regulates energy metabolism and is intrinsic to the self-renewal of hESCs. [ABSTRACT FROM AUTHOR]

High mortality (84–89%) of stocked rainbow trout (Oncorhynchus mykiss) in Twin Lakes, Washington, has been partially blamed on predation from non-native largemouth bass (Micropterus salmoides). We examined the gut-content of 434 largemouth bass and applied a bioenergetics model to determine the consumption demand on hatchery-released rainbow trout and other prey species in the Twin Lakes. Largemouth bass consumed approximately 4915 (95% CI; 2393–13,452) fall stocked rainbow trout in South Twin. No rainbow trout consumption was observed in North Twin, despite a similar stocking regime. Approximately 6.3% (95% CI; 3–17%) of total fall stocked rainbow trout in South Twin were consumed by largemouth bass. Rainbow trout stocked in the fall ranged from 100 to 160 mm total length and were all subject to predation by large largemouth bass ⩾300 mm. Large largemouth bass consumed rainbow trout that reached up to 210 mm in length and 58% of bass body length. No predation was observed on larger rainbow trout (215–370 mm) stocked during the spring. Smaller largemouth bass ⩽299 mm consumed primarily invertebrates, including crayfish, and did not consume rainbow trout in either lake. During spring and summer in South Twin Lake, large largemouth bass consumed principally golden shiner (Notemigonus crysoleucas) and crayfish. In North Twin Lake, golden shiner constituted most of the larger bass diet for the entire year. Differences in macrophyte distribution, bathymetry, temperature and/or predator-prey demographics likely influenced variability in largemouth bass predation between lakes. Largemouth bass predation did not account for overall rainbow trout mortality. [ABSTRACT FROM PUBLISHER]

Christensen, D.R. and B.C. Moore. 2009. Using stable isotopes and a multiple-source mixing model to evaluate fish dietary niches in a mesotrophic lake. Lake Reserv. Manage. 25:167–175. We used stable isotope analysis (SIA) of δ13C and δ15N and a multiple-source mixing model to evaluate dietary niches within the Twin Lakes, Washington, fish community to identify potential for exploitative competition, resource partitioning, and predation. The SIA revealed distinct spatial feeding niches; pelagic species were δ13C depleted, while littoral organisms were δ13C enriched. Trophic feeding niches were identified from δ15N enrichment with each successive trophic level. We found δ15N progression with increasing largemouth bass size, suggesting ontogenetic diet variability. Model results suggested that the smaller bass (i.e., ⩽299 mm length) fed principally on golden shiner, crayfish, and macroinvertebrates while larger bass (⩾300 mm) primarily consumed brook trout and golden shiner. Isotopic signatures for golden shiner were intermediary, indicating both pelagic and littoral feeding sources, including phantom midges (nocturnal and pelagic) and damselflies (littoral) as principal diet sources, suggesting possible horizontal diel migration in golden shiner. Pelagic zooplankton was indicated as the most important food source for rainbow trout. Food resources appeared to be partitioned among the Twin Lakes fish community, possibly limiting competitive interactions. Piscivory in largemouth bass was focused on golden shiner and brook trout, suggesting possible top-down regulation of lower trophic levels. Stable isotope analysis and multiple-source mixing models are useful tools that can improve lake and fishery management decisions by providing data on food web dynamics such as competition, resource partitioning, and predation in lakes. [ABSTRACT FROM PUBLISHER]

Sorsby fundus dystrophy (SFD) is an autosomal dominant macular dystrophy with an estimated prevalence of 1 in 220,000 and an onset of disease around the 4th to 6th decade of life. Similar to age-related macular degeneration (AMD), ophthalmoscopy reveals accumulation of protein/lipid deposits under the retinal pigment epithelium (RPE), referred to as drusen, in the eyes of patients with SFD. SFD is caused by variants in the gene for tissue inhibitor of metalloproteinases-3 (TIMP3), which has been found in drusen-like deposits of SFD patients. TIMP3 is constitutively expressed by RPE cells and, in healthy eyes, resides in Bruch's membrane. Most SFD-associated TIMP3 variants involve the gain or loss of a cysteine residue. This suggests the protein aberrantly forms intermolecular disulphide bonds, resulting in the formation of TIMP3 dimers. It has been demonstrated that SFD-associated TIMP3 variants are more resistant to turnover, which is thought to be a result of dimerisation and thought to explain the accumulation of TIMP3 in drusen-like deposits at the level of Bruch's membrane. An important function of TIMP3 within the outer retina is to regulate the thickness of Bruch's membrane. TIMP3 performs this function by inhibiting the activity of matrix metalloproteinases (MMPs), which have the function of catalysing breakdown of the extracellular matrix. TIMP3 has an additional function to inhibit vascular endothelial growth factor (VEGF) signalling and thereby to inhibit angiogenesis. However, it is unclear whether SFD-associated TIMP3 variant proteins retain these functions. In this review, we discuss the current understanding of the potential mechanisms underlying development of SFD and summarise all known SFD-associated TIMP3 variants. Cell culture models provide an invaluable way to study disease and identify potential treatments. These allow a greater understanding of RPE physiology and pathophysiology, including the ability to study the blood-retinal barrier as well as other RPE functions such as phagocytosis of photoreceptor outer segments. This review describes some examples of such recent in vitro studies and how they might provide new insights into degenerative diseases like SFD. Thus far, most studies on SFD have been performed using ARPE-19 cells or other, less suitable, cell-types. Now, induced pluripotent stem cell (iPSC) technologies allow the possibility to non-invasively collect somatic cells, such as dermal fibroblast cells and reprogram those to produce iPSCs. Subsequent differentiation of iPSCs can generate patient-derived RPE cells that carry the same disease-associated variant as RPE cells in the eyes of the patient. Use of these patient-derived RPE cells in novel cell culture systems should increase our understanding of how SFD and similar macular dystrophies develop., (Copyright © 2017 Elsevier Ltd. All rights reserved.)