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Kirkpatrick, J. F., & Turner, A. (2002). Reversibility of action and safety during pregnancy of immunization against porcine zona pellucida in wild mares (Equus caballus). Reprod Suppl, 60, 197–202.
Abstract: Contraceptive management of publicly valued wildlife species requires safeguards to ensure that these populations are preserved in a healthy state. In addition, reversibility of contraceptive effects and safety in pregnant animals are major concerns. A population of wild horses has been immunized against porcine zona pellucida (PZP) over a 12 year period on Assateague Island National Seashore, MD (ASIS). Mares initially received one or two 65 microg inoculations and once a year 65 microg booster inoculations, all delivered by dart. All young mares aged > 2 years were treated with PZP for 3 consecutive years regardless of whether they have bred successfully and they were then removed from treatment until they had foaled. All mares vaccinated for 1 or 2 consecutive years became fertile again and 69% of mares treated for 3 consecutive years returned to fertility. All five mares treated for 4 or 5 consecutive years have also returned to fertility, but over longer periods of time. Mares treated for 7 consecutive years have not returned to fertility, but several, while still infertile, have started ovulating again. There was no difference in survival rates between foals born to treated and untreated mares, and PZP treatment of pregnant mares did not affect subsequent fertility of their female offspring.
Keywords: Animals; *Animals, Wild; Antigens/administration & dosage; Contraception, Immunologic/methods/*veterinary; Egg Proteins/administration & dosage; Female; Fertility; *Horses; Immunization, Secondary/veterinary; Membrane Glycoproteins/administration & dosage; Population Control; Pregnancy; *Receptors, Cell Surface; Safety; Swine; Time Factors; Vaccines, Contraceptive/*administration & dosage
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Turner, A., & Kirkpatrick, J. F. (2002). Effects of immunocontraception on population, longevity and body condition in wild mares (Equus caballus). Reprod Suppl, 60, 187–195.
Abstract: Contraception is becoming a common approach for the management of captive and wild ungulates yet there are few data for contraceptive effects on entire populations. Management-level treatment of mares with porcine zona pellucida (PZP) vaccine resulted in zero population growth of the Assateague Island wild horse population within 1 year of initiation of treatment. Contraceptive efficacy was 90% for mares treated twice in the first year and annually thereafter. For mares given a single initial inoculation, contraceptive efficacy was 78%. The effort required to achieve zero population growth decreased, as 95, 83 and 84% of all adult mares were treated in each of the first 3 years, compared with 59 and 52% during the last 2 years. Mortality rates for mares and foals after the initiation of management-level treatments decreased below historic and pretreatment mortality rates of approximately 5%. Two new age classes have appeared among treated animals (21-25 years and > 25 years), indicating an increase in longevity among treated animals. Body condition scores for all horses, all adult mares and non-lactating mares increased significantly between summer 1989 and autumn 1999 but did not change significantly in lactating mares. These results provide reliable data for the construction of realistic models for contraceptive management of free-roaming or captive ungulate populations.
Keywords: Animals; *Animals, Wild; Antigens/administration & dosage; Body Constitution; Contraception, Immunologic/methods/*veterinary; Egg Proteins/administration & dosage; Female; *Horses; Longevity; Membrane Glycoproteins/administration & dosage; Population Control; Population Dynamics; *Receptors, Cell Surface; Swine; Vaccines, Contraceptive/administration & dosage
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Baltic, M., Jenni-Eiermann, S., Arlettaz, R., & Palme, R. (2005). A noninvasive technique to evaluate human-generated stress in the black grouse. Ann N Y Acad Sci, 1046, 81–95.
Abstract: The continuous development of tourism and related leisure activities is exerting an increasingly intense pressure on wildlife. In this study, a novel noninvasive method for measuring stress in the black grouse, an endangered, emblematic species of European ecosystems that is currently declining in several parts of its European range, is tested and physiologically validated. A radiometabolism study and an ACTH challenge test were performed on four captive black grouse (two of each sex) in order to get basic information about the metabolism and excretion of corticosterone and to find an appropriate enzyme-immunoassay (EIA) to measure its metabolites in the feces. Peak radioactivity in the droppings was detected within 1 to 2 hours. Injected (3)H-corticosterone was excreted as polar metabolites and by itself was almost absent. A cortisone-EIA was chosen from among seven tested EIAs for different groups of glucocorticoid metabolites, because it cross-reacted with some of the formed metabolites and best reflected the increase of excreted corticosterone metabolites, after the ACTH challenge test. Concentrations of the metabolites from fecal samples collected from snow burrows of free-ranging black grouse were within the same range as in captive birds. The noninvasive method described may be appropriate for evaluating the stress faced by free-living black grouse populations in the wild, particularly in mountain ecosystems where human disturbance, especially by winter sports, is of increasing conservation concern.
Keywords: Adrenocorticotropic Hormone/metabolism; Animals; Bird Diseases/*metabolism; Conservation of Natural Resources; Corticosterone/*metabolism; Ecosystem; Feces/*chemistry; Female; Galliformes/*metabolism; Immunoenzyme Techniques/methods/veterinary; Male; Reproducibility of Results; Stress/metabolism/*veterinary; Tritium/diagnostic use
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BASHORE, T. L., KEIPER, R., TURNER, J. W. J. R., & KIRKPATRICK J. F. (1990). The accuracy of fixed-wing aerial surveys of feral horses on a coastal barrier island. J. coast. res, 6, 53–56.
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Palme, R. (2005). Measuring fecal steroids: guidelines for practical application. Ann N Y Acad Sci, 1046, 75–80.
Abstract: During the past 20 years, measuring steroid hormone metabolites in fecal samples has become a widely appreciated technique, because it has proved to be a powerful, noninvasive tool that provides important information about an animal's endocrine status (adrenocortical activity and reproductive status). However, although sampling is relatively easy to perform and free of feedback, a careful consideration of various factors is necessary to achieve proper results that lead to sound conclusions. This article aims to provide guidelines for an adequate application of these techniques. It is meant as a checklist that addresses the main topics of concern, such as sample collection and storage, time delay extraction procedures, assay selection and validation, biological relevance, and some confounding factors. These issues are discussed briefly here and in more detail in other recent articles.
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Bartosova, J., Dvorakova, R., Vancatova, M., & Svobodova, I. (2008). Comprehension of human pointing gesture in domestic horses: Effect of training method. In IESM 2008.
Abstract: Horses have been considered to rely on human gesticular cues (McKinley and Sambrook 2000, Anim Cogn 3:13-22; and recently Maros et al. 2008, Anim Cogn 11:457-466), however large variability among individuals tested in two-ways object choice tasks was found. Part of the horses in those studies (40 and 26 %, respectively) even failed to pass adequately through the training session which preceded the testing phase and served to learn a horse to carry out a task. Therefore, we alternated the experimental design designed by McKinley and Sambrook (reduced number of testing trials to 10 per horse to keep its attention, applied just one, a dynamic-sustained pointing cue with touching the bucket, etc.), and tested an effect of training method, sex, age, and learning on proportion of correct choices. We hypothesised, that horses trained by “traditional” method (TTM) will get lower score than those experienced with “horsemanship-based” methods (HTM), being characterized by closer and more frequent human-horse contact and also extended exercising “from the ground” with frequent using of arms cues. Despite simplification of the methods, only about 60 % of tested horses passed through the training phase (i.e., learned to come to and upturn the bucket with hidden treat). Successful completion of training phase was reached regardless of age or sex of a horse, but by the training method; HTM horses ran better compared to TTM ones. No significant effect of age, sex, or learning (i.e., trial order within all 10), and training method as well was found on proportion of correct trials in the testing phase. Horses made a correct choice in more than 70% of trials. Individual scores ranged from 50 to 100 %. In conclusion, horses showed high level of comprehension of human pointing gesture, regardless of their sex or age. No effects of training method or learning process within a test suggest low impact of handling and learning on the level of comprehension at least of the most vivid human pointing gesture. Horses trained by methods based on “natural human-horse communication” did enhance cooperation with people.
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Kirkpatrick, J. F., Turner, J. W. J., Liu, I. K., Fayrer-Hosken, R., & Rutberg, A. T. (1997). Case studies in wildlife immunocontraception: wild and feral equids and white-tailed deer. Reprod Fertil Dev, 9(1), 105–110.
Abstract: Non-lethal management methods are required for wild equids that are protected by law and for deer inhabiting areas where lethal controls are not legal or safe. Single or multiple inoculations of porcine zona pellucida (PZP) vaccine have been delivered to wild horses and deer by means of darts. Contraceptive efficacy in horses after two inoculations ranged from 90% to 100%, and after a single inoculation ranged from 19% to 28%. Mares given a controlled-release form of the vaccine had foaling rates ranging from 7% to 20%. No detectable changes in social organization or behaviours among treated horses occurred. Contraceptive effects were reversible after 4 consecutive years of treatment but 5-7 years of treatment resulted in ovulation failure and decreased urinary oestrogen concentrations. Among deer, two inoculations were 70-100% effective in preventing fawns, but one inoculation yielded a contraceptive efficacy of < or = 20%, with pregnancies occurring late in the breeding season; a single annual booster inoculation reduced fertility to 20% in the second year. Energy costs of extended breeding seasons were less than those resulting from pregnancy. After two years of treatment, ovaries appeared normal. These studies suggest that PZP immunocontraception can be successfully applied to certain free-roaming populations of wild horses and deer.
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Turner, J. W. J., Liu, I. K., & Kirkpatrick, J. F. (1996). Remotely delivered immunocontraception in free-roaming feral burros (Equus asinus). J Reprod Fertil, 107(1), 31–35.
Abstract: Regulation of local overpopulations of free-roaming feral equids is in demand worldwide for ecological balance and habitat preservation. Contraceptive vaccines have proven effective in feral horses, which breed seasonally, but no data are available for equids such as the burro, which is reproductively active all year round. In the present study, 27 individually identified female feral burros (Equus asinus) roaming free in Virgin Islands National Park (St John, US Virgin Islands; Lesser Antilles) were remotely treated with pig zonae pellucidae (PZP) vaccine. Between January and May, 16 burros were darted with a 1 ml emulsion of PZP plus Freund's adjuvant. Ten to twelve months later each treated burro was given a single booster injection of PZP plus adjuvant to maintain contraception through a second year. Eleven adult untreated jennies served as controls. Beginning one year after initial vaccination, these burros were monitored for pregnancy and foal production. Collection of data to determine treatment effect was not begun until 12 months after initial treatment to ensure that pregnancies existing before vaccination were not included. Pregnancy was assessed using previously validated methods for steroid metabolite measurement in fresh faecal samples. None of the PZP-treated burros produced foals between 0 and 12 months after the last inoculation. One PZP-treated burro tested positive for pregnancy at 10 months after the final inoculation. During this same period, six of 11 untreated burros tested pregnancy-positive, and four were observed with foals. There was no difference in pregnancy rates among treated, control and randomly sampled jennies between 12 and 24 months after the last inoculation. The results demonstrate that, in free-roaming feral burros that are reproductively active all year round: (1) burros can be accessed for remotely delivered PZP vaccination; (2) PZP contraception is effective; (3) PZP contraception is reversible; and (4) pregnancy can be reliably detected by faecal steroid analysis.
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Rosa, P. A. J., Azevedo, A. M., & Aires-Barros, M. R. (2007). Application of central composite design to the optimisation of aqueous two-phase extraction of human antibodies. J Chromatogr A, 1141(1), 50–60.
Abstract: The partition of human antibodies in aqueous two-phase systems (ATPSs) of polyethylene glycol (PEG) and phosphate was systematically studied using first pure proteins systems and then an artificial mixture of proteins containing 1mg/ml human immunoglobulin G (IgG), 10mg/ml serum albumin and 2mg/ml myoglobin. Preliminary results obtained using pure proteins systems indicated that the PEG molecular weight and concentration, the pH value and the salts concentration had a pronounced effect on the partitioning behaviour of all proteins. For high ionic strengths and pH values higher than the isoelectric point (pI) of the contaminant proteins, IgG could be selectively recovered on the top phase. According to these results, a face centred composite design was performed in order to optimise the purification of IgG from the mixture of proteins. The optimal conditions for the isolation of IgG were observed for high concentrations of NaCl and low concentrations of both phase forming components. The best purification was achieved using an ATPS containing 8% (w/w) PEG 3350, 10% (w/w) phosphate pH 6 and 15% (w/w) NaCl. A recovery yield of 101+/-7%, a purity of 99+/-0% and a yield of native IgG of 97+/-4% were obtained. Back extraction studies of IgG to a new phosphate phase were performed and higher yields were obtained using 10% phosphate buffer at pH 6. The total extraction yield was 76% and the purity 100%.
Keywords: Analysis of Variance; Animals; Antibodies/*chemistry/*isolation & purification; Buffers; Chemical Fractionation/*methods; Horses; Humans; Hydrophobicity; Isoelectric Point; Models, Biological; Molecular Weight; Myoglobin/chemistry/isolation & purification; Osmolar Concentration; Phase Transition; Polyethylene Glycols; Serum Albumin/chemistry/isolation & purification; Sodium Chloride
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Kaiser, L., Heleski, C. R., Siegford, J., & Smith, K. A. (2006). Stress-related behaviors among horses used in a therapeutic riding program. J Am Vet Med Assoc, 228(1), 39–45.
Abstract: OBJECTIVE: To determine whether therapeutic riding resulted in higher levels of stress or frustration for horses than did recreational riding and whether therapeutic riding with at-risk individuals was more stressful for the horses than was therapeutic riding with individuals with physical or emotional handicaps. DESIGN: Observational study. ANIMALS: 14 horses in a therapeutic riding program. PROCEDURE: An ethogram of equine behaviors was created, and horses were observed while ridden by 5 groups of riders (recreational riders, physically handicapped riders, psychologically handicapped riders, at risk children, and special education children). Number of stress-related behaviors (ears pinned back, head raised, head turned, head tossed, head shaken, head down, and defecation) was compared among groups. RESULTS: No significant differences in mean number of stress-related behaviors were found when horses were ridden by recreational riders, physically handicapped riders, psychologically handicapped riders, or special education children. However, mean number of stress-related behaviors was significantly higher when horses were ridden by the at-risk children. CONCLUSIONS AND CLINICAL RELEVANCE: Results suggest that for horses in a therapeutic riding program, being ridden by physically or psychologically handicapped individuals is no more stressful for the horses than is being ridden in the same setting by recreational riders. However, at-risk children caused more stress to the horses, suggesting that the time horses are ridden by at-risk children should be limited both daily and weekly.
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