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Different dental schools assess 'finals' in different ways. Assessment of applied clinical knowledge and skills are commonly based on either objective structured clinical examinations or multiple 'long cases'. While iterative changes naturally occur throughout education and assessment, it is important to step back and question what skills should be assessed and either identify or develop new assessment methods. At the point of graduation, these should capture the professional attributes expected of a 'qualified dentist'. New graduate attributes we consider valuable, which can often be challenging to assess summatively in a high-stakes examination, include effective communication skills in a variety of contexts, the ability to be reactive and responsive, and to manage case complexity appropriately for the graduate level.Here, we report a dynamic assessment incorporating candidate interaction with a range of stakeholders. To reflect progression of changing clinical situations, we move away from solely traditional case-based discussions, which principally assess interaction with an examiner, and have introduced multiple scenarios with rapid situation changes, long-term follow-up, treatment complications and challenging communications. A cross-cutting assessment rubric was developed to assess candidates' information gathering/giving, communication, clinical and diagnostic reasoning, applied knowledge, management and professionalism., Competing Interests: Ethics declaration. The authors have no conflicts of interest to declare. This manuscript is an education case study; therefore, ethical approval was not required., (© 2024. The Author(s).)

Introduction: Active involvement of patients in healthcare professional education is well established, taking a variety of forms. There is a steer towards patient feedback informing the development of dental students and while there is recognition of its potential value to individual students, challenges exist related to collection and use. What is unclear is, within a dental education setting, the extent and use of patient feedback to individual students. A scoping review was conducted to assess and map the volume and characteristics of the research/literature in this area., Methods: Systematic searches of bibliographic databases Ovid MEDLINE(R), Scopus, ERIC and Embase were conducted, and wider literature (Google Scholar) was searched. Screening was conducted based on eligibility criteria and a customised data charting form was used., Results: The electronic and citation tracking searches identified 1021 studies. After duplicates were removed, 778 studies were screened by title and abstract, and 718 studies were found to be irrelevant to the current review. Sixty full-text studies were assessed for eligibility, 46 studies were excluded, and 14 studies were included for data charting., Conclusion: This review has identified that patient feedback has been captured through both simulated and real patient encounters. There was a bias towards feedback generated through simulated patient encounters. Feedback was reported to support the development of a range of skills, most frequently communication and patient management. Challenges that were identified by researchers related to staff/student engagement and available resources., (© 2024 The Author(s). European Journal of Dental Education published by John Wiley & Sons Ltd.)

Objective: To evaluate pharmacy, dental and dental therapy undergraduate students' perceived competence of interprofessional working before and after attending an interprofessional education (IPE) Gerodontology workshop., Background: Whilst there is international recognition of the importance of collaboration between the dental profession and systemic healthcare providers to enhance patient care, there remains a paucity of research into IPE in Gerodontology., Materials and Methods: Pharmacy, dental and dental therapy undergraduate students attended a 2-hour Gerodontology case-based workshop. Students completed anonymised Interprofessional Collaborative Competencies Attainment Surveys (ICCAS) before and after attendance., Results: 108 questionnaires were received, 7 were withdrawn (1 incomplete with only pre-workshop side completed, 6 did not identify degree programme). From 101 included questionnaires, 37 were from pharmacy, 56 dental and 8 dental therapy students resulting in response rates of 84%, 82% and 67%, respectively. Each student group recorded an increase in positive reflective competence median (IQR) after taking part in the workshop. Overall median (IQR) reflective competence before the workshop was 6 (1), 5 (2) and 6 (2) for pharmacy, dental and dental therapy students, respectively, which increased to 7 (1) for all groups. There was variability in reflective competence before attending the workshop between dental and pharmacy students for two questions, and dental and dental therapy students for two different questions., Conclusion: All students reported increased reflective competence of interprofessional working following the workshop. These findings suggest that introduction of IPE events into Gerodontology curricula may improve student understanding and appreciation of interprofessional working when providing care for older people., (© 2022 The Authors. Gerodontology published by Gerodontology Association and John Wiley & Sons Ltd.)

Eighty percent of the estimated 600 million domestic cats in the world are free-roaming. These cats typically experience suboptimal welfare and inflict high levels of predation on wildlife. Additionally, euthanasia of healthy animals in overpopulated shelters raises ethical considerations. While surgical sterilization is the mainstay of pet population control, there is a need for efficient, safe, and cost-effective permanent contraception alternatives. Herein, we report evidence that a single intramuscular treatment with an adeno-associated viral vector delivering an anti-Müllerian hormone transgene produces long-term contraception in the domestic cat. Treated females are followed for over two years, during which transgene expression, anti-transgene antibodies, and reproductive hormones are monitored. Mating behavior and reproductive success are measured during two mating studies. Here we show that ectopic expression of anti-Müllerian hormone does not impair sex steroids nor estrous cycling, but prevents breeding-induced ovulation, resulting in safe and durable contraception in the female domestic cat., (© 2023. The Author(s).)

The impacts of urbanization on bird biodiversity depend on human-environment interactions that drive land management. Although a commonly studied group, less attention has been given to public perceptions of birds close to home, which can capture people's direct, everyday experiences with urban biodiversity. Here, we used ecological and social survey data collected in the metropolitan region of Phoenix, Arizona, USA, to determine how species traits are related to people's perceptions of local bird communities. We used a trait-based approach to classify birds by attributes that may influence human-bird interactions, including color, size, foraging strata, diet, song, and cultural niche space based on popularity and geographic specificity. Our classification scheme using hierarchical clustering identified four trait categories, labeled as Metropolitan (gray, loud, seedeaters foraging low to ground), Familiar (yellow/brown generalist species commonly present in suburban areas), Distinctive (species with distinguishing appearance and song), and Hummingbird (hummingbird species, small and colorful). Strongly held beliefs about positive or negative traits were also more consistent than ambivalent ones. The belief that birds were colorful and unique to the regional desert environment was particularly important in fortifying perceptions. People largely perceived hummingbird species and birds with distinctive traits positively. Similarly, urban-dwelling birds from the metropolitan trait group were related to negative perceptions, probably due to human-wildlife conflict. Differences arose across sociodemographics (including income, age, education, and Hispanic/Latinx identity), but explained a relatively low amount of variation in perceptions compared with the bird traits present in the neighborhood. Our results highlight how distinctive aesthetics, especially color and song, as well as traits related to foraging and diet drive perceptions. Increasing people's direct experiences with iconic species tied to the region and species with distinguishing attributes has the potential to improve public perceptions and strengthen support for broader conservation initiatives in and beyond urban ecosystems., (© 2022 The Ecological Society of America.)

Urbanization is one of the most widespread and extreme examples of habitat alteration. As humans dominate landscapes, they introduce novel elements into environments, including artificial light, noise pollution, and anthropogenic food sources. One understudied form of anthropogenic food is refuse from restaurants, which can alter wildlife populations and, in turn, entire wildlife communities by providing a novel and stable food source. Using data from the Maricopa Association of Governments and the Central Arizona-Phoenix Long Term Ecological Research (CAP LTER) project, we investigated whether and how the distribution of restaurants influences avian communities. The research aimed to identify restaurants, and thus the associated food they may provide, as the driver of potential patterns by controlling for other influences of urbanization, including land cover and the total number of businesses. Using generalized linear mixed models, we tested whether the number of restaurants within 1 km of bird monitoring locations predict avian community richness and abundance and individual species abundance and occurrence patterns. Results indicate that restaurants may decrease avian species diversity and increase overall abundance. Additionally, restaurants may be a significant predictor of the overall abundance of urban-exploiting species, including rock pigeon (Columba livia), mourning dove (Zenaida macroura), and Inca dove (Columbina Inca). Understanding how birds utilize anthropogenic food sources can inform possible conservation or wildlife management practices. As this study highlights only correlations, we suggest further experimental work to address the physiological ramifications of consuming anthropogenic foods provided by restaurants and studies to quantify how frequently anthropogenic food sources are used compared to naturally occurring sources., Competing Interests: The authors have declared that no competing interests exist.

Ecosystems are defined, studied, and managed according to boundaries constructed to conceptualize patterns of interest at a certain scale and scope. The distinction between ecosystems becomes obscured when resources from multiple origins cross porous boundaries and are assimilated into food webs through repeated trophic transfers. Ecosystem compartments can define bounded localities in a heterogeneous landscape that simultaneously retain and exchange energy in the form of organic matter. Here we developed and tested a framework to quantify reciprocal reliance on cross-boundary resource exchange and calculate the contribution of primary production from adjacent ecosystem compartments cycling through food webs to support consumers at different trophic levels. Under this framework, an integrated ecosystem can be measured and designated when the boundary between spatially distinct compartments is permeable and the bidirectional exchange of resources contributes significantly to sustaining both food webs. Using a desert river and riparian zone as a case study, we demonstrate that resources exchanged across the aquatic-riparian boundary cycle through multiple trophic levels. Furthermore, predators on both sides of the boundary were supported by externally produced resources to a similar extent, indicating this is a tightly integrated river-riparian ecosystem and that changes to either compartment will substantially impact the other. Using published data on lake ecosystems, we demonstrated that benthic and pelagic ecosystem compartments are likely not fully integrated, but differences between lakes could be used to test ecological hypotheses. Finally, we discuss how the integrated ecosystem framework could be applied in urban-preserve and field-forest ecosystems to address a broad range of ecological concepts. Because few systems function in complete isolation, this novel approach has application to research and management strategies globally as ecosystems continue to face novel pressures that precipitate cascading ecological repercussions well beyond a bounded system of focus., (© 2021 by the Ecological Society of America.)

Felid semen has historically been frozen using an egg yolk-based cryopreservation medium. However, the use of egg introduces several potential concerns, such as variability in composition, microbial contamination, and regulatory issues. In the present study, our aim was to compare a chemically-defined, soy-based medium (SOY) to a commercial egg yolk-based medium (TEY) for the cryopreservation of sperm in four imperiled small cat species. Semen was collected from adult male cats (n = 6 black-footed cats; n = 6 sand cats; n = 4 fishing cats; and n = 7 Pallas' cats) via electroejaculation, split into two aliquots, and cryopreserved in SOY or TEY. Frozen-thawed samples were evaluated for sperm motility and rate of progressive motility (up to 24 h post-thaw) and acrosome status (0 and 6 h). No difference in post-thaw traits were observed between treatments in all four species. Heterologous IVF using oocytes collected laparoscopically from domestic cats demonstrated no difference among freezing treatments in percentage of mature oocytes that cleaved or the mean number of blastomeres at 48 h post-insemination. More spermatozoa frozen with SOY were bound to the zona pellucida in the sand cat (P = 0.018), but no treatment effect was observed in the other three species. These findings collectively demonstrate that SOY may be a preferable alternative to TEY for sperm cryopreservation in these four wild felid species., Competing Interests: Declaration of competing interest The authors have no competing interests to declare., (Copyright © 2020 Elsevier Inc. All rights reserved.)

Chediak-Higashi Syndrome (CHS) is a well-characterized, autosomal recessively inherited lysosomal disease caused by mutations in lysosomal trafficking regulator (LYST). The feline model for CHS was originally maintained for ~20 years. However, the colonies were disbanded and the CHS cat model was lost to the research community before the causative mutation was identified. To resurrect the cat model, semen was collected and cryopreserved from a lone, fertile,  CHS carrier male. Using cryopreserved semen, laparoscopic oviductal artificial insemination was performed on three queens, two queens produced 11 viable kittens. To identify the causative mutation, a fibroblast cell line, derived from an affected cat from the original colony, was whole genome sequenced. Visual inspection of the sequence data identified a candidate causal variant as a ~20 kb tandem duplication within LYST, spanning exons 30 through to 38 (NM_001290242.1:c.8347-2422_9548 + 1749dup). PCR genotyping of the produced offspring demonstrated three individuals inherited the mutant allele from the CHS carrier male. This study demonstrated the successful use of cryopreservation and assisted reproduction to maintain and resurrect biomedical models and has defined the variant causing Chediak-Higashi syndrome in the domestic cat.

Dental education providers have raised concerns around the challenges experienced when trying to demonstrate student attainment of 'professionalism', as described in the General Dental Council (GDC) document 'Preparing for practice'. This manuscript describes a documentary analysis of each 'outcome' within the dentist professionalism domain section, with an assessment of whether it met the criteria of a functional learning outcome. In addition, outcomes were scrutinised in terms of application within undergraduate degree programmes. Key challenges identified were: the 'outcome list' included standards and objectives as well as outcomes; not all statements were specific and focused, some were broad and multi-faceted; determining attainment where there was no identifiable endpoints; and judging achievability within the supervised and confined environment of undergraduate programmes. We conclude that issues have been created through the way that these learning outcomes have been expressed. Also, it is evident that there are ideological and philosophical issues about accurately articulating the attainment of complex phenomena like professionalism. The findings from this study reflect the challenges associated with using learning outcomes to adequately capture professionalism, the values that underpin it and the behaviours that manifest its existence in clinical practice.

Propagation of giant river otters (GRO) in zoos is inconsistent: some pairs never reproduce while others are prolific in producing young but can be hindered by low cub survival. Developing effective breeding programs requires understanding normal reproductive parameters and behavior. Fecal samples were collected for 6-16 months from five breeding pairs, two individual females, and one female pair at seven zoos, and analyzed for fecal progesterone, estrogen, testosterone, and glucocorticoid (FGM) metabolites via enzyme immunoassay. Enclosure characteristics and management routines were recorded at six facilities where behavior was assessed over 1 week. Median fecal progestogens during pregnancy and pseudopregnancy were ∼2.5-3.8× greater than basal concentrations. Gestation lasted 66.5 ± 3.5 days (62-70 days); pseudopregnancies lasted 58 ± 11.6 days (41-69 days). Elevated progestogens indicate ovulation but cannot distinguish pregnancy from pseudopregnancy. Periodically sustained, elevated progestogens observed in two females housed without a male indicated spontaneous ovulation. Elevations in fecal estrogens were not associated with estrus, and seasonality in male testosterone was not observed. Wavering scream and contact call vocalizations among reproductively successful males and females, respectively, suggested the importance of social communication. Most facilities housing successful pairs had larger enclosures with more water than land area, vegetation, and limited public exposure. Baseline FGM were negatively correlated with enclosure size and percentage of water area (p < 0.05), and lower baseline FGM were associated with reproductive success (p < 0.05). These results suggest that housing GRO in spacious enclosures with open water and some insulation from disturbance might promote appropriate behavior, lower FGM, and reproduction., (© 2018 Wiley Periodicals, Inc.)

Introduction: Outcome-based programmes provide a framework to support educators and learners in understanding content and end points within taught courses. Management of these outcomes in the Dental Degree at Newcastle University has been a challenge in relation to quality assurance and enhancement, having over 1500 detailed student-level outcomes (SLO). This research aimed to explore the implications of adopting a more superficial "course" level of outcome (CLO), when reviewed against a reference set of external LO requirements., Materials and Methods: A purposive sample of five courses within the undergraduate dental programme was selected. The mapping of both SLOs and CLOs was reviewed in relation to their total number and the mapping connections to the reference outcomes., Results: There was a mean reduction of 79% in outcomes when comparing SLOs to CLOs. The number of mapping connections between CLOs and the reference set reduced in three courses and increased in two, when compared to SLOs., Discussion: From a purely numerical perspective, changing the detail of learning outcomes has led to a change in mapping connections. As the delivered curriculum has remained unchanged, this demonstrates a potential impact of differing interpretations of learning outcomes. Further review of learning outcomes in relation to the domains categorised within the reference outcome document suggested more mapping links were obtained in clinically focused courses than academic or theoretical courses., Conclusion: A demonstrable impact in mapping connections was observed when the detail within the learning outcomes was changed. This has implications for programme leaders in structuring LOs for a curriculum., (© 2016 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.)

Species conservation requires a thorough understanding of habitat requirements. The northern Mexican gartersnake (Thamnophis eques megalops) was listed as threatened under the U.S. Endangered Species Act in 2014. Natural resource managers are interested in understanding the ecology of this subspecies to guide management decisions and to determine what features are necessary for habitat creation and restoration. Our objective was to identify habitat selection of northern Mexican gartersnakes in a highly managed, constructed wetland hatchery. We deployed transmitters on 42 individual gartersnakes and documented use of habitat types and selection of specific habitat features. Habitat selection was similar between males and females and varied seasonally. During the active season (March-October), gartersnakes primarily selected wetland edge habitat with abundant cover. Gestating females selected similar locations but with less dense cover. During the inactive season (November-February), gartersnakes selected upland habitats, including rocky slopes with abundant vegetation. These results of this study can help inform management of the subspecies, particularly in human-influenced habitats. Conservation of this subspecies should incorporate a landscape-level approach that includes abundant wetland edge habitat with a mosaic of dense cover for protection and sparsely vegetated areas for basking connected to terrestrial uplands for overwintering.

Anthropogenic water sources (AWS) are developed water sources used as a management tool for desert wildlife species. Studies documenting the effects of AWS are often focused on game species; whereas, the effects on non-target wildlife are less understood. We used live trapping techniques to investigate rodent abundance, biomass, and diversity metrics near AWS and paired control sites; we sampled vegetation to determine rodent-habitat associations in the Sauceda Mountains of the Sonoran Desert in Arizona. A total of 370 individual mammals representing three genera and eight species were captured in 4,800 trap nights from winter 2011 to spring 2012. A multi-response permutation procedure was used to identify differences in small mammal community abundance and biomass by season and treatment. Rodent abundance, biomass, and richness were greater at AWS compared to control sites. Patterns of abundance and biomass were driven by the desert pocket mouse ( Chaetodipus penicillatus ) which was the most common capture and two times more numerous at AWS compared to controls. Vegetation characteristics, explored using principal components analysis, were similar between AWS and controls. Two species that prefer vegetation structure, Bailey's pocket mouse ( C. baileyi ) and white-throated woodrat ( Neotoma albigula) , had greater abundances and biomass near AWS and were associated with habitat having high cactus density. Although small mammals do not drink free-water, perhaps higher abundances of some species of desert rodents at AWS could be related to artificial structure associated with construction or other resources. Compared to the 30-year average of precipitation for the area, the period of our study occurred during a dry winter. During dry periods, perhaps AWS provide resources to rodents related to moisture., Competing Interests: The authors declare there are no competing interests.

Semen cryopreservation and storage in genome resource banks (GRBs), in combination with artificial insemination (AI), could be invaluable for genetic management and conservation of endangered otter species. For any applied conservation benefit, effective methods for otter sperm processing and cryopreservation first must be established. In this study, our objective was to develop an effective semen cryopreservation method for the North American river otter, evaluating the effect of extender composition (i.e., glycerol concentration, Equex STM paste supplementation) and freezing protocol (timing of glycerol addition, pre-freeze cooling rate, freezing/packaging method) on post-thaw sperm motility, longevity and acrosome status. Semen was collected from 14 otters housed at 9 zoos, and following cryopreservation in an egg-yolk based extender, thawed to assess sperm motility and acrosome status immediately post-thaw and during 6 h of in vitro culture. Results indicated that extender containing 4% glycerol was preferable (p < 0.05) to 8% glycerol but the temperature/timing of extender addition containing 4% glycerol did not affect (p > 0.05) post-thaw sperm parameters. Treatments with extender containing Equex and frozen by pelleting on dry ice showed greater (p < 0.05) motility and percentage of intact acrosomes compared to treatments frozen in extender without Equex, regardless of pre-freeze cooling rate. In the absence of Equex, pelleting provided superior post-thaw sperm motility (p < 0.01) and higher (p < 0.001) percentage of sperm with intact acrosomes compared to samples frozen in straws over liquid nitrogen vapor. Results of this study indicate that cryopreservation of otter sperm using an egg-yolk -TEST based extender containing 4% glycerol and 1% Equex, with the pellet freezing method, provided superior post-thaw sperm motility, longevity and acrosomal integrity compared to other combinations. Neither alterations in timing of glycerolated extender addition nor pre-freeze cooling rate had a discernable effect on post-thaw otter sperm parameters. These findings represent the first assessment of semen cryopreservation in any otter species and may be of value as a model for development of semen cryopreservation strategies in other endangered otter species., (Copyright © 2017 Elsevier Inc. All rights reserved.)

Semen banking of domestic cats and wild felids represents a vital resource for their long-term conservation, but current methods require access to advanced training and specialized equipment. A newer method of semen collection, urethral catheterization of medetomidine-treated cats, allows recovery of high sperm numbers, but it is unclear if this approach permits maximal sperm recovery or is feasible using less expensive alpha-2 agonists. Similarly, a newer sperm preservation approach, vitrification, offers advantages of simplicity and minimal equipment needs, but its efficacy in combination with urethral catheterization has not been investigated. Our specific objectives were to (i) evaluate sequential semen recovery with urethral catheterization and electroejaculation in domestic cats, (ii) assess the effectiveness of a weak (xylazine) versus strong (dexmedetomidine) alpha-2 agonist for inducing sperm release, and (iii) compare post-thaw sperm motility, acrosome status and fertilizing capacity of catheter-recovered samples after vitrification or straw freezing. Results indicated that electroejaculation following repeated catheterization allowed recovery of additional spermatozoa (range, 11-32 × 10 6  sperm/male) and that xylazine was ineffective for inducing meaningful sperm release (range, 0-0.4 × 10 6 sperm/male). Post-thaw motility and acrosome status of vitrified catheter samples did not differ (p > .05) from that of straw frozen samples. Preliminary results indicated that in vitro fertilization success (9/30, 30%) of vitrified catheter sperm did not differ (p > .05) from that observed with straw frozen samples (17/30, 57%). In conclusion, urethral catheterization of dexmedetomidine-treated cats allows recovery of substantial sperm numbers but electroejaculation still may be warranted for maximal sperm recovery. Xylazine is not suitable as an inexpensive alternative to dexmedetomidine for catheterization. Vitrification of catheter samples results in comparable post-thaw parameters to straw freezing and may be adequate for use with oviductal insemination procedures., (© 2016 Blackwell Verlag GmbH.)

Measurement of circulating luteinizing hormone (LH) concentrations in cats and temporal changes following ovariohysterectomy (OHE) or possibly GnRH vaccination may be informative for assessing their fertility, contraception or sterilization status. In this study, serum LH concentrations were measured in domestic cats (n = 6) immediately prior to and up to 120 days post-OHE. Basal LH concentrations of females previously subjected to OHE (n = 4; ~1.5 years post-OHE) were compared pre- and post-vaccination with a GnRH immunocontraceptive, and to LH concentrations in intact females. Basal serum LH concentrations (2.67 ± 0.43 ng/ml; mean ± SEM) in intact females increased (p < .01) by 30 days post-OHE (5.65 ± 0.87 ng/ml) but then declined (p < .05) to pre-OHE levels (mean range, 3.26-3.62 ng/ml) at days 60-120 post-OHE. Serum LH (3.84 ± 0.51 ng/ml) in four females ~1.5 years after OHE tended to be higher (p = .10) than those of intact females prior to OHE. Three months following first or second GnRH immunocontraceptive vaccine treatment, serum LH values in females previously subjected to OHE decreased (p < .05) to concentrations similar to those observed in intact females. Our preliminary results suggest that OHE of domestic cats causes a marked increase in basal LH levels within the first few weeks after ovariohysterectomy followed by a return to pre-OHE basal values over the next several months. Reduced LH concentrations after GnRH vaccine may indicate the effectiveness of the immunocontraceptive in reducing the circulating levels of GnRH, thereby reducing secretion of LH., (© 2016 Blackwell Verlag GmbH.)

Introduction: Providers of education programmes are required to demonstrate to students, staff and external regulators the content of the programme and how this aligns to key outcomes. Whilst many programme themes are straightforward to illustrate, other integral themes prove challenging to demonstrate independently., Innovation: A virtual course takes elements within the current programme relating to a specific theme, repackages and maps them to provide clear signposting by highlighting each occasion the theme is taught, experienced and assessed. The individual elements remain within their current course, so there is no disruption to the established curriculum, but there is an enhancement in terms of transparently displaying linkages within a theme., Discussion: Using the concept of virtual courses we have found we can respond to new initiatives and requirements of external regulators, as well as providing improved signposting for staff and students. This can be undertaken without the need to redesign a whole curriculum., Conclusion: In well-designed and currently effective curricula, we would like to recommend that virtual course development can enhance programme transparency, demonstrate external regulatory requirements and promote quality assurance without disruption to current content., (© 2015 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.)

Artificial insemination (AI) in cats traditionally uses equine chorionic gonadotropin (eCG) and human chorionic gonadotropin (hCG) to induce follicular development and ovulation, with subsequent bilateral laparoscopic intrauterine insemination. However, long-acting hCG generates undesirable secondary ovulations in cats. Uterine AI also requires relatively high numbers of spermatozoa for fertilization (~8 × 10(6) sperm), and unfortunately, sperm recovery from felids is frequently poor. Using short-acting porcine luteinizing hormone (pLH) instead of hCG, and using the oviduct as the site of sperm deposition, could improve fertilization success while requiring fewer spermatozoa. Our objectives were to compare pregnancy and fertilization success between 1) uterine and oviductal inseminations and 2) eCG/hCG and eCG/pLH regimens in domestic cats. Sixteen females received either eCG (100 IU)/hCG (75 IU) or eCG (100 IU)/pLH (1000 IU). All females ovulated and were inseminated in one uterine horn and the contralateral oviduct using fresh semen (1 × 10(6) motile sperm/site) from a different male for each site. Pregnant females (11/16; 69%) were spayed approximately 20 days post-AI, and fetal paternity was genetically determined. The number of corpora lutea (CL) at AI was similar between hormone regimens, but hCG increased the number of CL at 20 days post-AI. Numbers of pregnancies and normal fetuses were similar between regimens. Implantation abnormalities were observed in the hCG group only. Finally, oviductal AI produced more fetuses than uterine AI. In summary, laparoscopic oviductal AI with low sperm numbers in eCG/hCG- or eCG/pLH-treated females resulted in high pregnancy and fertilization percentages in domestic cats. Our subsequent successes with oviductal AI in eCG/pLH-treated nondomestic felids to produce healthy offspring supports cross-species applicability.

The objective was to compare a proprietary egg yolk-based cryopreservation medium with a chemically defined soy-based medium, as well as to examine effects of temperature of glycerol addition on sperm parameters and IVF after freezing and thawing of domestic cat sperm. Semen was collected from adult cats (four males and three ejaculates per male), divided in four equal aliquots, and extended in either egg yolk with 4% glycerol added before (EYG) or after (EY) cooling to 5 °C, or soy-lecithin with 4% glycerol added before (SLG) or after (SL) cooling to 5 °C. Extended sperm were frozen in straws over liquid nitrogen vapor. Sperm progressive motility (%) and rate of progressive movement (scale of 0-5) were evaluated at 0, 1, 3, 6, and 24 h post-thaw. Sperm capacitation, acrosome integrity, and DNA integrity were assessed at 15 min post-thaw. Effects of media (EY or SL) on IVF success was also examined (three males and three ejaculates per male). Sperm motility was greater (P < 0.05) in soy-based compared with egg yolk-based media at 3, 6, and 24 h post-thaw. A higher (P < 0.05) percentage of noncapacitated sperm (pattern F) were present in soy-based (SLG, 63.7 ± 9.2%; and SL, 64.1 ± 9.2%) compared with egg yolk-based (EYG, 49.9 ± 7.9%; and EY, 52.4 ± 18.6%) cryopreservation media, regardless of temperature of glycerol addition. Addition of glycerol at 5 °C increased (P < 0.05) percentage of sperm motility at 6 h (EYG 16.3 ± 8.3% vs. EY, 24.0 ± 11.7%; SLG, 36.7 ± 6.5% vs. SL, 42.9 ± 10.1%) and 24 h (EYG, 2.1 ± 3.3% vs. EY, 8.3 ± 3.9%; SLG, 11.3 ± 8.3% vs. SL, 18.8 ± 7.4%) post-thaw in both media. There were no differences (P > 0.05) between cryodiluents in embryo cleavage, percentage of embryos reaching blastocyst, or cell number per blastocyst. The chemically defined, soy-based medium resulted in better preservation of long-term motility and capacitation status of frozen-thawed domestic cat sperm compared with a commercial egg yolk-based extender, without compromising fertilizing ability., (Copyright © 2012 Elsevier Inc. All rights reserved.)

Artificial insemination (AI) is potentially invaluable as an adjunct to natural breeding for the conservation management of non-domestic felid populations. The efficacy of AI, however, must be substantially improved for applied use, especially when using frozen semen. Our recent advances in using laparoscopic oviductal AI (LO-AI) with low sperm numbers and freezing of cat semen in a soy lecithin-based cryoprotectant medium suggest that combining these two approaches might improve pregnancy outcomes with frozen-thawed spermatozoa. In this study, our objectives were to (i) assess the effect of two gonadotropin dosages (100 vs 150 IU eCG) on ovarian response in domestic cats and (ii) compare the relative fertility of frozen-thawed and fresh semen in vivo following LO-AI. All 16 females ovulated after gonadotropin treatment and were inseminated with fresh semen from one male and frozen-thawed semen from a second male. There were no differences between gonadotropin dosages in CL number, pregnancy percentage or litter size. Half (8/16) of the females conceived, with seven females giving birth to a total of 36 offspring. Paternity analysis showed that more kittens resulted from LO-AI with fresh (28/36, 78%) than frozen-thawed (8/36, 22%) semen, possibly due to impaired motility and longevity of thawed sperm. These results demonstrated that viable offspring can be produced by AI using semen frozen in a soy lecithin-based medium. Insemination with greater numbers of frozen-thawed spermatozoa, combined with further refinement of cat sperm cryopreservation methods, may be necessary to optimize pregnancy success with LO-AI in domestic and nondomestic cats., (© 2012 Blackwell Verlag GmbH.)

Increasing human populations have resulted in aggressive water development in arid regions. This development typically results in altered stream flow regimes, reduced annual flow volumes, changes in fluvial disturbance regimes, changes in groundwater levels, and subsequent shifts in ecological patterns and processes. Balancing human demands for water with environmental requirements to maintain functioning ecosystems requires quantitative linkages between water in streams and ecosystem attributes. Streams in the Sonoran Desert provide important habitat for vertebrate species, including resident and migratory birds. Habitat structure, food, and nest-building materials, which are concentrated in riparian areas, are provided directly or indirectly by vegetation. We measured riparian vegetation, groundwater and surface water, habitat structure, and bird occurrence along Cherry Creek, a perennial tributary of the Salt River in central Arizona, USA. The purpose of this work was to develop an integrated model of groundwater-vegetation-habitat structure and bird occurrence by: (1) characterizing structural and provisioning attributes of riparian vegetation through developing a bird habitat index (BHI), (2) validating the utility of our BHI through relating it to measured bird community composition, (3) determining the riparian plant species that best explain the variability in BHI, (4) developing predictive models that link important riparian species to fluvial disturbance and groundwater availability along an arid-land stream, and (5) simulating the effects of changes in flow regime and groundwater levels and determining their consequences for riparian bird communities. Riparian forest and shrubland vegetation cover types were correctly classified in 83% of observations as a function of fluvial disturbance and depth to water table. Groundwater decline and decreased magnitude of fluvial disturbance caused significant shifts in riparian cover types from riparian forest to shrublands. Variability in the BHI was best explained by the cover of deciduous riparian tree species, primarily Populus fremontii, Platanus wrightii, and Salix gooddingii. The distributions of these plant species were well explained by the depth to groundwater and magnitude of fluvial disturbance along the stream. Bird species diversity and richness were significantly higher in sites with higher habitat indices. This quantitative linkage between surface and groundwater, plant species composition, habitat complexity, and bird communities has implications for water management and in determining environmental flows.

A study was conducted opportunistically to evaluate the potential of rescuing immature oocytes from the ovaries of the Sumatran rhinoceros postmortem. Recovered oocytes (n = 30) were placed in maturation culture for 36 hr and inseminated with frozen-thawed homologous spermatozoa. After culture, evaluation of nuclear maturation status revealed that a large number of oocytes were degenerated (n = 21), but nine oocytes were assessed at the germinal vesicle (n = 3), metaphase I (n = 3), and metaphase II (n = 3) stages. Frozen-thawed Sumatran rhinoceros spermatozoa were capable of binding to the zona pellucida of in vitro matured oocytes, but no fertilization or cleavage resulted. In conclusion, relatively large numbers of oocytes can be obtained by ovarian follicular aspiration postmortem in the Sumatran rhinoceros, and some of these oocytes are capable of achieving nuclear maturation in vitro. However, additional studies are required to improve maturation success and achieve fertilization in culture.

In this study, fecal samples were collected from 24 North American river (NARO) and 17 Asian small-clawed otters (ASCO) for 6-36 months and semen collected seasonally from NARO males (n=4/season) via electroejaculation. Our main objectives were to: (1) characterize endocrine parameters by longitudinal monitoring of fecal hormone metabolites and (2) investigate semen collection and basal seminal traits in NARO. NARO demonstrated a distinct seasonality in the spring, with females having a monoestrual estrogen elevation lasting 15.33+/-1.98 (mean+/-SEM) days and males peaking in testosterone production for 25.50+/-7.51 days. Pregnancy was characterized by 7-9 months of basal fecal progesterone, presumably corresponding to embryonic diapause, followed by a rapid increase over the final 68-73 days to term. Pseudopregnancy exhibited a similar late winter progesterone peak of 68-72 days, which could not be differentiated from pregnancy. Geographic latitude possibly influenced the timing of increased testosterone in males and increased progesterone in pregnant/pseudopregnant females. In ASCO, monitoring of fecal estrogens did not allow consistent detection of peak values associated with behavioral estrus. Both pregnancy and pseudopregnancy were characterized by a moderate rise in fecal progesterone for 14-16 days postovulation followed by a marked increase. Total gestation length was 67-77 days compared with 62-84 days for pseudopregnancy. In NARO, optimal sperm recovery and quality occurred only in the spring, corresponding with seasonal increases in testicular volume and fecal testosterone. These findings represent the first comprehensive information on normative endocrine and seminal traits in freshwater otter species.

It is well established that estrogen administration during neonatal development can advance pubertal onset and prevent the maintenance of regular estrous cycles in female rats. This treatment paradigm also eliminates the preovulatory rise of gonadotropin releasing hormone (GnRH). It remains unclear, however, through which of the two primary forms of the estrogen receptor (ERalpha or ERbeta) this effect is mediated. It is also unclear whether endocrine disrupting compounds (EDCs) can produce similar effects. Here we compared the effect of neonatal exposure to estradiol benzoate (EB), the ERalpha specific agonist 1,3,5-tris(4-hydroxyphenyl)-4-propyl-1H-pyrazole (PPT), the ERbeta specific agonist diarylpropionitrile (DPN) and the naturally occurring EDCs genistein (GEN) and equol (EQ) on pubertal onset, estrous cyclicity, GnRH activation, and kisspeptin content in the anteroventral periventricular (AVPV) and arcuate (ARC) nuclei. Vaginal opening was significantly advanced by EB and GEN. By 10 weeks post-puberty, irregular estrous cycles were observed in all groups except the control group. GnRH activation, as measured by the percentage of immunopositive GnRH neurons that were also immunopositive for Fos, was significantly lower in all treatment groups except the DPN group compared to the control group. GnRH activation was absent in the PPT group. These data suggest that neonatal exposure to EDCs can suppress GnRH activity in adulthood, and that ERalpha plays a pivotal role in this process. Kisspeptins (KISS) have recently been characterized to be potent stimulators of GnRH secretion. Therefore we quantified the density of KISS immunolabeled fibers in the AVPV and ARC. In the AVPV, KISS fiber density was significantly lower in the EB and GEN groups compared to the control group but only in the EB and PPT groups in the ARC. The data suggest that decreased stimulation of GnRH neurons by KISS could be a mechanism by which EDCs can impair female reproductive function.

Endocrine active compounds (EACs) have been shown to influence a number of reproductive endpoints but less is known about how they might affect other hormone dependent behaviors including anxiety and aggression. Recent evidence suggests that these effects may be mediated through the beta form of the estrogen receptor (ERbeta). Using male Long Evans rats, we sought to determine how neonatal exposure to EACs affects anxiety and aggression in adulthood. Anxiety was assessed using the elevated plus maze and aggression was assessed 8 weeks later using the resident intruder test. To gain insight into which ER subtype (ERalpha vs ERbeta) might be mediating these effects we used agonists specific for ERalpha (1,3,5-tris(4-Hydroxyphenyl)-4-propyl-1H-pyrazole (PPT)) or ERbeta (Diarylpropionitrile (DPN)) as additional treatment groups. For these experiments the synthetic EAC bisphenol-A (BPA) and the phytoestrogen metabolite equol (EQ) were used. Male neonates were injected with either 0.05 ml sesame oil (control), 50 microg estradiol benzoate (EB), 1 mg/kg DPN, 1 mg/kg PPT, 50 microg/kg BPA, or 10 mg/kg EQ daily for 4 days beginning on the day of birth (PND 0). Compared to the oil treated controls, significantly fewer open arm entries were made by the males neonatally treated with DPN, EQ, or BPA. The DPN and EQ treated males were also more aggressive compared to the controls. These findings suggest that neonatal exposure to EACs with agonistic activity on ERbeta may influence affective behavior in adulthood, including anxiety and aggression.

The objective of this study was to define the physiologic needs of domestic cat embryos to facilitate development of a feline-specific culture medium. In a series of factorial experiments, in vivo-matured oocytes (n = 2040) from gonadotropin-treated domestic cats were inseminated in vitro to generate embryos (n = 1464) for culture. In the initial study, concentrations of NaCl (100.0 vs. 120.0 mM), KCl (4.0 vs. 8.0 mM), KH(2)PO(4) (0.25 vs. 1.0 mM), and the ratio of CaCl(2) to MgSO(4)-7H(2)O (1.0:2.0 mM vs. 2.0:1.0 mM) in the medium were evaluated during Days 1-6 (Day 0: oocyte recovery and in vitro fertilization [IVF]) of culture. Subsequent experiments assessed the effects of varying concentrations of carbohydrate (glucose, 1.5, 3.0, or 6.0 mM; l-lactate, 3.0, 6.0, or 12.0 mM; and pyruvate, 0.1 or 1.0 mM) and essential amino acids (EAAs; 0, 0.5, or 1.0x) in the medium during Days 1-3 and Days 3-6 of culture. Inclusion of vitamins (0 vs. 1.0x) and fetal calf serum (FCS; 0 vs. 5% [v/v]) in the medium also was evaluated during Days 3-6. Development and metabolism of IVF embryos on Day 3 or Day 6 were compared to age-matched in vivo embryos recovered from naturally mated queens. A feline-optimized culture medium (FOCM) was formulated based on these results (100.0 mM NaCl, 8.0 mM KCl, 1.0 mM KH(2)PO(4), 2.0 mM CaCl(2), 1.0 mM MgSO(4), 1.5 mM glucose, 6.0 mM L-lactate, 0.1 mM pyruvate, and 0x EAAs with 25.0 mM NaHCO(3), 1.0 mM alanyl-glutamine, 0.1 mM taurine, and 1.0x nonessential amino acids) with 0.4% (w/v) BSA from Days 0-3 and 5% FCS from Days 3-6. Using this medium, ~70% of cleaved embryos developed into blastocysts with profiles of carbohydrate metabolism similar to in vivo embryos. Our results suggest that feline embryos have stage-specific responses to carbohydrates and are sensitive to EAAs but are still reliant on one or more unidentified components of FCS for optimal blastocyst development.

Cryopreservation of spermatozoa from free-living ocelots (Leopardus pardalis) could benefit their conservation by facilitating gene flow between in situ and ex situ populations without requiring removal of additional cats from the wild. The objective of this study was to investigate three different methods of ocelot sperm cryopreservation to identify the most appropriate technique for use in a field environment. Male ocelots (n = 10), housed in North American zoos, were anaesthetised with tiletamine-zolazepam (7 mg kg(-1) bodyweight; i.m.) and subjected to a regimented electroejaculation procedure. Recovered semen was evaluated for sperm concentration, motility and morphology and processed for cryopreservation by three methods: (1) pelleting on dry ice, (2) freezing in straws over liquid nitrogen vapour; and (3) freezing in straws in a dry shipper. Frozen samples were thawed and assessed for post-thaw acrosome status, viability, motility over time and ability to fertilize viable domestic cat oocytes. Although several post-thaw sperm parameters varied (P < 0.05) among freezing methods, frozen-thawed ocelot spermatozoa from all treatments showed a similar (P > 0.05) capacity to bind, penetrate and fertilize viable domestic cat oocytes. These findings suggest that spermatozoa collected from male ocelots under field conditions may be frozen in straws either using liquid nitrogen alone or in a charged dry shipper to retain adequate functional competence after thawing for use with assisted reproductive procedures.

High doses of estradiol (E(2)) can impair spatial learning in the Morris water maze, in ovariectomized mice, but the same dose has no effect on adult castrated males. Here, we test the hypothesis that this sex difference is caused by neonatal actions of E(2). In Experiment 1, C57BL/6J pups were given daily estradiol benzoate (EB) or oil injections from the day of birth until postnatal Day 3. Adults were gonadectomized and received EB (s.c.) or oil 28 h before the first day of training, and 4 h before each of four daily training sessions on the Morris water maze. Females given oil as neonates, and EB prior to training displayed the poorest performance. Females that received EB as neonates and EB prior to training were insensitive to the deleterious effects of adult EB and performed better than males given the same hormone treatments. We conducted a second experiment using aromatase enzyme knockout (ArKO) mice. Adult male and female ArKO and wild-type (WT) littermates were gonadectomized and received either injections of oil or EB prior to and during water maze training (as described above). Hormone treatment failed to affect performance, yet, female but not male ArKO mice showed impaired learning compared to WT littermates. Thus, exposure to estradiol during neonatal development can counteract the deleterious effects of EB on adult spatial learning.

Although herpesviruses are known to contaminate the semen of several mammalian species, the occurrence of feline herpesvirus type 1 (FHV-1) in semen of infected cats has not been reported. Our objectives in this study were to investigate the presence of FHV-1 DNA in seminal fluid and frozen-thawed spermatozoa from FHV-1 infected Pallas' cats (Otocolobus manul) and assess the functionality of their frozen-thawed spermatozoa in vitro. Over a 3-yr period, semen (n = 33 ejaculates) was collected periodically via electroejaculation from four Pallas' cats chronically infected with FHV-1. Spermic ejaculates were frozen by pelleting on dry ice and stored in liquid nitrogen. After thawing, sperm motility and acrosome status were assessed over time during in vitro culture. For vitro fertilization (IVF), viable domestic cat (Felis silvestris catus) oocytes were inseminated with frozen-thawed Pallas' cat spermatozoa and evaluated for embryo cleavage. For FHV-1 polymerase chain reaction (PCR) analysis, DNA was extracted from seminal fluid, frozen-thawed spermatozoa, inseminated oocytes, heterologous IVF embryos, and conjunctival biopsies and analyzed for presence of a 322-base pair region of the FHV-1 thymidine kinase gene. Immediately post-thaw, sperm motility and percentage of intact acrosomes were decreased (P < 0.05) compared to fresh samples, and declined further (P < 0.05) during culture. However, all frozen-thawed IVF samples were capable of fertilizing domestic cat oocytes (overall, 46.1 +/- 6.0% cleavage). PCR analysis did not identify FHV-1 DNA in any reproductive sample despite the repeated detection of FHV-1 DNA in conjunctival biopsies. These results suggest that semen collected from Pallas' cats infected with FHV-1 does not contain cell-associated or non-cell-associated virus and that frozen-thawed spermatozoa exhibit adequate function for potential genetic rescue with minimal risk of FHV-1 transmission.

One approach to study interactions between behavior and genetics is to use inbred mice with different genetic backgrounds. To examine the effect of background on a specific gene, we conducted a series of experiments with a well-characterized knockout (KO) mouse, the estrogen receptor alpha KO (ERalphaKO). The ERalphaKO mouse has so far been examined in one inbred line, C57BL/6J. Here, we examined the behavior of ERalphaKO mice within three different backgrounds mixed with C57BL/6J; DBA/2J, BALB/c, and A/J. First, we assessed masculine sexual behavior in both intact male and testosterone-treated female offspring. More ERalphaKO males in the DBA/2J (5/12) and BALB/c (5/13) backcrosses displayed intromissions and many ejaculated as compared with males in a C57BL/6J and A/J mixed background. Many fewer ERalphaKO females than males displayed masculine sexual behavior in any of the three hybrid crosses. We assessed fertility in males from the C57BL/6J by DBA/2J cross and found that one of 12 ERalphaKO males sired a litter. Several other characteristics of sexual behavior and physiology were unaffected by genetic background in ERalphaKO mice. Our data suggest that genetic background has dramatic effects on male sexual behavior and its dependence on the ERalpha gene.

A current priority for the preservation of the endangered red wolf (Canis rufus) is the development of a sperm-based genome resource bank. The aims of this study were to examine the effects of (i) holding temperature on the motility of spermatozoa over time, and (ii) cooling methods on the characteristics of spermatozoa after cooling and cryopreservation. Electroejaculates (n = 11; fresh) were evaluated for the percentage of motile spermatozoa, cell and acrosome morphology (Spermac (Meditech 1st Canada Inc, Montreal, Ontario) and fluorescein isothiocyanate-labelled Pisum sativum agglutinin lectin (PSA/FITC; Sigma Diagnostics, Oakville, Ontario) staining), and zona penetration. Semen samples were then divided into two equal samples and centrifuged to remove seminal plasma. One half of the ejaculate sample was re-suspended in sperm-Tyrode's albumin lactate pyruvate (TALP), divided into three aliquots and maintained either at room temperature (approximately 21-23 degrees C), 0 degree C or 37 degrees C. Sperm motility was examined at 0.5 and 1.0 h, and subsequently every hour for 10 h. Motility of spermatozoa decreased after 2 h, but was consistently greater at room temperature than at 37 degrees C or 0 degree C. The other half of the ejaculate sample was re-suspended in an egg yolk-based extender and divided into two aliquots. One aliquot was cooled in a refrigerator (5 degrees C) for 30 min, whereas the second aliquot was put into a beaker containing water at 37 degrees C, which was then placed into an ice bath until the sample reached 0 degree C (approximately 120 min). Spermatozoa were evaluated after cooling and after freezing and thawing treatments. No differences were observed between cooling treatments either after cooling or freezing and thawing. However, marked decreases in intact acrosomes, post-thaw motility and normal morphology of spermatozoa after treatment demonstrate that further investigations are necessary to improve cryopreservation methods in this species.

Recent advances in feline and canine reproductive studies demonstrate how methodically piecing this information together is beginning to reap rewards for wildlife conservation programs. Non-invasive endocrinology can be used to monitor female reproductive function, time con-specific introductions or AI, and diagnose pregnancy. Sperm morphology characteristics and cell membrane function may be genetically inherited and differ between genetically diverse and inbred species/populations in felids. It is not clear if the same is true for the endangered red wolf. While standards exist for freezing feline and canine sperm, new information using fluorescent staining and zona penetration assays (ZPA) indicates that significant damage can occur during pre-freeze cooling, and may also be related to a species' genetic diversity. Posthumous gamete salvage from genetically valuable animals not only provides a means to study sperm and oocyte physiology but also to assist with genetic management of populations. Using the knowledge gained, IVM/IVF and ICSI have been successful in the domestic cat and AI has resulted in offspring in numerous non-domestic felids. However, understanding the processes of IVM/IVF is still not well understood in canids. New information reveals that sperm and the cumulus cells may be integral to oocyte maturation and that canine epididymal sperm are not capable of undergoing fertilization. The acquisition of knowledge and application of biotechnologies lags behind for non-domestic canid conservation programs.