The influence of selenium and vitamin E supplementation on cytological assessment of red blood cell line of bone marrow in fallow deer kept in captivity.

Cytological evaluation of bone marrow smears stained by May-Grünwald Giemsa method was performed. The smears came from 20 fallow deer (Dama dama) 3 days old divided into 2 groups each consisting of 10 animals. The experimental group (E) received intramuscularly selenium and vitamin E at a dose of 3.0 ml (tocopherol acetate - 50 mg, sodium selenite - 0.5 mg, solvent - 1 ml) in the 3rd day of age. The control group (C) did not receive any supplementation or placebo. For hematological analyzes blood was collected three times: on 0, 15th and 25th day of the experiment. Serum concentration of selenium and vitamin E was determined using high perfor- mance liquid chromatography and glutathione peroxidase activity (GSH-Px) by kinetic method. On the 15th day after supplementation, a statistically significant increase in the percentage of erythroblastic cell line was observed in bone marrow smears. At that time, the increase in GSH-Px activity in the E group was also observed, reaching the value of 165.3 U/gHb, which was statisti- cally significant. The percentage of proerythroblasts (8.23% in group E and 5.02% in group C) differed significantly between groups at the 25th day after supplementation. This study revealed that supplementation of selenium and vitamin E resulted in an increase in the number of erythro- cytes to an average of 13.5 (˟ 10¹²/l) in the experimental group on 25th day with a significant increase in hemoglobin to 193 g/l in the experimental group.

Circannual changes in serum concentrations of thyroxine, calcitonin and parathormone in immature and mature red deer females (Cervus elaphus).

The aim of this study was to determine circannual changes in the serum concentrations of thyroxine, calcitonin and parathormone in mature and immature red deer females. Blood samples from 8 hinds were collected monthly for 26 months. Secretions of thyroxine and calcitonin showed circannual rhythms with significantly higher levels in the immature hinds compared to the mature animals (p<0.05). For thyroxine, the concentration was higher in the winter/spring period than in summer/autumn (p<0.05), while for calcitonin the concentration profile was the opposite (p<0.05). The concentration of parathormone was significantly higher in summer/autumn that in the other months of the experiment (p<0.01). These results may indicate that the hormones investigated may be involved in the regulation of seasonal reproductive activity and in processes contributing to entering puberty in red deer females.

Cadmium, lead, and mercury exposure assessment among croatian consumers of free-living game.

Free-living game can be an important source of dietary cadmium and lead; the question is whether exposure to these two elements is such that it might cause adverse health effects in the consumers. The aim of this study was to estimate dietary exposure to cadmium, lead, and mercury from free-living big game (fallow deer, roe deer, red deer, wild boar, and brown bear), and to mercury from small game (pheasant and hare), hunted in Croatia from 1990 to 2012. The exposure assessment was based on available literature data and our own measurements of metal levels in the tissues of the game, by taking into account different consumption frequencies (four times a year, once a month and once a week). Exposure was expressed as percentage of (provisional) tolerable weekly intake [(P)TWI] values set by the European Food Safety Authority (EFSA). Consumption of game meat (0.002-0.5 % PTWI) and liver (0.005-6 % PTWI) assumed for the general population (four times a year) does not pose a health risk to consumers from the general population, nor does monthly (0.02-6 % PTWI) and weekly (0.1-24 % PTWI) consumption of game meat. However, because of the high percentage of free-living game liver and kidney samples exceeding the legislative limits for cadmium (2-99 %) and lead (1-82 %), people should keep the consumption of certain game species' offal as low as possible. Children and pregnant and lactating women should avoid eating game offal altogether. Free-living game liver could be an important source of cadmium if consumed on a monthly basis (3-74 % TWI), and if consumed weekly (11-297 % TWI), it could even give rise to toxicological concern.

Post-mortem hemoparasite detection in free-living Brazilian brown brocket deer (Mazama gouazoubira, Fischer 1814) / Detecção post-mortem de hemoparasitos em veados catingueiro (Mazama gouazoubira, Fischer 1814) de vida livre no Brasil

Tick-borne infections can result in serious health problems for wild ruminants, and some of these infectious agents can be considered zoonosis. The aim of the present study was the post-mortem detection of hemoparasites in free-living Mazama gouazoubira from Minas Gerais state, Brazil. The deer samples consisted of free-living M. gouazoubira (n = 9) individuals that died after capture. Necropsy examinations of the carcasses were performed to search for macroscopic alterations. Organ samples were collected for subsequent imprint slides, and nested PCR assays were performed to detect hemoparasite species. Imprint slide assays from four deer showed erythrocytes infected with Piroplasmida small trophozoites, and A. marginale corpuscles were observed in erythrocytes from two animals. A. marginale and trophozoite co-infections occurred in two deer. A nested PCR analysis of the organs showed that six of the nine samples were positive for Theileria sp., five were positive for A. phagocytophilum and three were positive for A. marginale, with co-infection occurring in four deer. The results of the present study demonstrate that post-mortem diagnostics using imprint slides and molecular assays are an effective method for detecting hemoparasites in organs.

Patógenos transmitidos por carrapatos podem resultar em sérios problemas de saúde para os ruminantes selvagens, e alguns podem ser zoonoses. O objetivo do presente estudo foi a detecção post mortem de hemoparasitos, em Mazama gouazoubira de vida livre, oriundos de Minas Gerais, através da análise de lâminas de impressão e nested PCR. Foram amostrados nove M. gouazoubira de vida livre, que morreram após a captura. Exames de necropsia foram realizados, e as carcaças foram examinadas para detectar alterações macroscópicas. Amostras dos órgãos foram coletadas para a realização de imprint em lâminas e para nested PCR à procura de hemoparasitos. A análise das lâminas mostrou pequenos trofozoítos de Piriplasmida nos eritrócitos de quatro dos oito animais examinados, e corpúsculos de Anaplasma marginale foram observados nos eritrócitos de dois dos cervídeos. A coinfecção com A. marginale e trofozoítos de piroplasmas ocorreu em dois animais. As análises de nPCR dos órgãos mostraram que seis dos nove animais estavam positivos para Theileria sp., cinco para A. phagocytophilum e três para A. marginale, sendo que a coinfecção ocorreu em quatro cervídeos. Os resultados do presente estudo demonstram que os diagnósticos post-mortem, pelas imprints em lâminas e ensaios moleculares, são métodos eficazes de detecção de hemoparasitos nos principais órgãos parasitados.

Plasma progesterone changes and length of oestrous cycle in Rusa Deer (Rusa timorensis).

A study was conducted to profile the plasma progesterone (P4) concentrations and establish the length of oestrous cycle in the Rusa timorensis during the breeding season. Five healthy hinds were selected for peripheral blood sampling twice weekly to gauge the P4 levels by radioimmunoassay, at the start of the breeding season indicated by rutting behaviours of sexually active males. The hinds were polyestrous as proven by the cyclic trend of P4 levels. After the presumptive oestrus indicated by the lowest P4 concentrations (0.20±0.09ng/ml), this ovarian hormone was markedly elevated on day 7 of the cycle (0.78±0.20ng/ml), reached its peak (2.61±0.23ng/ml, P<0.05) on day 14, and then declined to the basal level in the subsequent oestrus. The mean oestrous cycle length in R. timorensis during the breeding season was 19.2 days with a range of 18-21 days, and the pattern of circulating progesterone during the oestrous cycle of the R. timorensis is similar to those of other deer species. It was also observed that the length of oestrous cycle of R. timorensis determined by gauging the progesterone levels and observation of the oestrous behaviours as well as changes in the cellular pattern of vaginal epithelial cells are highly consistent.

Seasonal luteal cyclicity of pubertal and adult red deer (Cervus elaphus).

Reproductive failure of rising-two-year-old (R(2)) hinds and seasonal misalignment between calving and pastoral feed production are two factors limiting reproductive productivity of farmed red deer hinds in New Zealand. This study aimed to better understand processes around female puberty and breeding seasonality by describing the potential breeding season (i.e., oestrous cyclicity) of three red deer genotypes. A total of 27 hinds born in December 2005, representing Eastern European (Cervus elaphus hippelaphus), Western European (C.e. scoticus) and F1 crossbred (C.e. hippelaphus×scoticus) red deer, were blood sampled thrice-weekly for 7-8 months (February-September/October) across two years spanning the potential breeding seasons as R(2)'s in 2007 (i.e., puberty) and as adults in 2008. Plasma progesterone profiles were used to construct breeding cycle histories for each hind. Four R(2) hinds failed to initiate oestrous cycles (i.e., puberty failure). The remaining R(2) hinds, including all F1 hinds, exhibited between two and seven oestrous cycles. F1 hinds were significantly earlier to initiate, and later to terminate, cyclic activity, resulting in a longer mean pubertal breeding season (139 days) than for Eastern (86 days) and Western hinds (86 days). However, the data for R(2) hinds are confounded by live-weight, with the F1 hinds being significantly heavier than other genotypes. There were significant correlations between live-weight and seasonality parameters in 2007. All hinds were cyclic as adults in 2008, exhibiting between four and nine oestrous cycles, and a mean breeding season duration of between 132 (Western) and 137 (F1) days. For adult hinds there were no significant genotype differences in cyclic onset and cessation timing, and no observable relationships between live-weight and any reproductive parameter. However, the mean dates for the onset of the breeding season for all genotypes in 2008 were 2-3 weeks later than normally expected for adult hinds in New Zealand. The reasons for this are unclear but may relate to chronic stress of frequent animal handling. The study has demonstrated that puberty in red deer hinds is associated with a shorter potential breeding season than for adult hinds, and that perturbation of breeding activity appears to be quite common, leading to incidences of puberty failure and possibly other aberrant cyclic events. Live-weight×genotype interactions may influence puberty but do not appear to be strongly expressed in adults. However, the relatively late onset of oestrous cyclicity in the adult hinds may be an artefact of the study that has masked genetic influences on seasonal breeding patterns.

Biochemical variables in free-ranging white-tailed deer (Odocoileus virginianus) after chemical immobilization in clover traps or via ground-darting.

The objective of this prospective observational cohort study in free-ranging female white-tailed deer (Odocoileus virginianus) was to compare the physiologic effects of two methods of anesthetic drug administration: hand-injection in Clover traps and remote injection by dart after ground-stalking. Six trapped and 14 darted female deer were injected with a median (minimum, maximum) of 590 microg/kg butorphanol (401, 1070 microg/kg), plus 235 microg/kg medetomidine (160, 429 microg/kg) intramuscularly. In the trap, the deer struggled when approached and were restrained for injection. Darted deer sprinted away after injection. Once immobilized, deer were transported to a veterinary hospital where blood was collected and vital signs were measured on admission. Admission data from a subset of deer in which measurements were taken within 40 min of trapping (n = 6) or darting (n = 5) were analyzed. After salpingectomy under isoflurane and while still anesthetized, another blood sample was collected from all 20 deer. Body weight and immobilization drug doses were not different between groups. On admission, most deer from both groups were hypoxemic, although the darted deer were significantly more hypoxemic. The median rectal temperature in trapped deer was higher than in darted deer, and temperatures higher than 39 degrees C only occurred in trapped deer. The median heart rate in trapped deer was more than twice that in darted deer. Trapped deer had lower median pH and base excess; in trapped deer, the median plasma lactate concentration was more than fivefold higher than in darted deer. After surgery, the median serum creatine kinase concentration was nearly 10-fold higher in trapped deer, and the median cardiac troponin I concentration was higher in trapped deer but undetectable in 10 of 14 darted deer. The white-tailed deer immobilized by hand-injection in Clover traps experienced more severe physiologic perturbations than deer remotely injected by dart after ground-stalking. These perturbations might be sufficient to cause myocardial damage.

Relationship between rank and plasma testosterone and cortisol in red deer males (Cervus elaphus).

The objective of this study was to assess the effect of a change in the social composition in a group of red deer males on the relationship between their rank and testosterone. A group of twelve adult red deer males (Cervus elaphus) was tested in two social settings. From April 15 to June 9 (Period 1) this group was kept separately in an enclosure. On June 10, nine 3-year-old males were added to that group of adult males. They were kept together until August 31. We performed 10 observations of the group when the agonistic interactions of the males were recorded and we took 9 blood samples per male in Period 1; 11 observations were made and 10 samples were taken in Period 2. Concentrations of testosterone and cortisol were later determined in plasma. Adding much younger and smaller sparring partners into the experimental group of adult males in Period 2 altered the agonistic behaviour of the adults even though this did not trigger any change in rank position of the experimental males except one. Adult males targeted preferentially their attacks on individuals much lower in the hierarchy. Experimental male deer with higher social rank had lower levels of testosterone in Period 1; in Period 2 it was just the opposite. In Period 1 the animals had higher cortisol levels than in Period 2. As controls we used four adult (5years old) males sharing the enclosure with four 3-year-old males. No changes in hormone concentrations were observed in the control group. Thus, changing the social environment of adult red deer males resulted in change of the relationship between rank and testosterone and cortisol concentrations.

Analysis of the value of measurement of the activity of caeruloplasmin as an alternative to measurement of the concentration of elemental copper in plasma and serum of farmed red deer.

AIM: To evaluate the relationship between the concentration of elemental Cu and activity of caeruloplasmin (CP) in the plasma and serum of farmed red deer (Cervus elaphus). METHODS: Paired serum and heparinised plasma samples taken from 142 deer from 10 herds in the North Island of New Zealand were tested for the concentration of elemental Cu and activity of CP. Additionally, liver biopsy samples were taken from 49 of the deer (all one herd). The relationship between individual results for the concentration of elemental Cu and activity of CP was examined using a segmented model, with random slope and intercept and threshold. RESULTS: The activity of CP in plasma and that in serum were strongly correlated (R2=0.994), with a high level of agreement. The concentration of Cu and activity of CP also showed a strong association (R2>0.88 for both plasma and serum), although there were significant differences between herds, with the segmented regression model providing the best fit rather than a linear mixed model. Using the current thresholds for elemental Cu, the relationship between the concentration of Cu and activity of CP suggested that an activity of CP of <4 IU/L indicates Cu deficiency, and that deer with an activity of CP of >or=7 IU/L have adequate Cu status. CONCLUSIONS: The relationship between elemental Cu and CP is markedly different from the simple linear relationship previously reported for cattle. Above a threshold of 10 IU/L, the rise in elemental Cu per unit increase in activity of CP was 50% of that below 10 IU/L. This is probably because above this threshold the oxidative activity of CP per enzyme unit was greater. CLINICAL RELEVANCE: The association between concentration of Cu and activity of CP was robust enough to define provisional thresholds for determining Cu status. However, the proximity of the deficiency threshold to the limit of detection of the CP test, the effect of herd on the association, and the inherent disadvantages of using an enzymatic system, mean that activity of CP should not be used routinely to estimate Cu status in deer. When assessing the Cu status of deer based on blood samples, the concentration of Cu in plasma or serum should be used.

Brief report: a new profile of terminal N-acetyllactosamines glycans on pig red blood cells and different expression of alpha-galactose on Sika deer red blood cells and nucleated cells.

It has been reported that: (1) large variations were found in the number of sialic acid (SA) capped with N-acetyllactosamines (SA-Galbeta1-4GlcNAc-R) and alpha-Gal epitopes (Galalpha1-3Galbeta1-4GlcNAc-R) or uncapped N-acetyllactosamines (Galbeta1-4GlcNAc-R) on different mammalian red blood cells, and on nucleated cells originating from a given tissue in various species; (2) goat, sheep, horse and mouse red blood cells lack alpha-Gal epitopes, despite the expression of this epitope on a variety of nucleated cells in these species, including lymphocytes differentiated from the same hematopoietic origin. In this study, flow cytometry and Western blot analyses of pig red blood cells showed that alpha-Gal epitopes on pig red cells developed concomitantly after treatment with neuraminidase, suggesting that the terminal N-acetyllactosaminide glycans were capped with SA-alpha-Gal epitopes. Whereas, the expression of the alpha-Gal epitopes on red blood cells from Sika deer (Cevus nippon hortulorum) were found to be absent even though the epitopes were present on their white blood cells. Thus, these results add new data not only for the terminal carbohydrate structures on cell surface glycans of various mammalian cells, but also for wide variety of epitope expression on the cells from different tissues, which might be useful for understanding their unique states resulting from differentiation and evolution.

Testosterone, but not IGF-1, LH, prolactin or cortisol, may serve as antler-stimulating hormone in red deer stags (Cervus elaphus).

The role of androgens and insulin-like growth factor 1 (IGF-1) in antler growth has been disputed. We predicted that the secretory of IGF-1 may be associated with an acceleration of body growth rather than with antler growth. Furthermore we anticipated a relationship between the increase of testosterone and the progress of antler growth. If IGF-1 is involved in the stimulation of antler growth, this should be more obvious in young than in mature stags. Eight two-year-old red deer stags (Cervus elaphus), and twelve adult red deer stags were blood sampled and the length of their velvet antlers was measured in one-week intervals during the period of antler growth. Concentrations of testosterone, cortisol, IGF-1, luteinizing hormone (LH), and prolactin were determined in plasma by enzyme immunoassay or radioimmunoassay. Antler growth per day was primarily dependent on changes in testosterone concentration per day in both groups of stags. As expected, only in two-year-old stags we detected a possible role of IGF-1 in the antler growth regulation, but that was not in agreement with previously published studies. Nevertheless, this effect was still utilized in interaction with testosterone. In addition to total antler length, only concentrations of testosterone and LH were significantly higher in adult males in comparison to two-year-old males. Our present results lead us to conclude that it is not IGF-1 but testosterone which is responsible for the intensity of antler growth in subadult and adult red deer stags.

Proteinograma sérico de veados-catingueiro (Mazama gouazoubira) criados em cativeiro obtido por eletroforese em gel de agarose e de poliacrilamida (SDS PAGE) / Serum protein concentrations of captive brown brocket deer (Mazama gouazoubira) determined by means of agarosis and sodium dodecyl sulphate-polyacrylamide (SDS-PAGE) gel electrophoresis

The serum protein concentrations of brown brocket deer (Mazama gouazoubira) obtained by agarosis gel and sodium dodecyl sulphate-polyacrylamide (SDS-PAGE) gel were determined from blood samples of ten adult healthy animals (six females and four males), monthly collected in the morning, during 12 months. The animals, maintained in individual stable and protected from noise, received ad libitum a diet of comercial ration and green roughage. Serum protein concentrations in agarosis gel revealed the presence of four protein fractions: albumin, alphaglobulin, betaglobulin, and gammaglobulin. Only serum concentrations of albumin were influenced by season, being values in spring higher than values in summer (4.15 x 3.64g/dl). Serum concentrations of albumin (4.05 x 3.75g/dl) were higher for female and alphaglobulin (0.39 x 0.53g/dl) werehigher for males. Results showed 34 proteins with molecular weights ranging from 18kD to 165kD. Significant differences between at least two seasons were found on values of 11 proteins. In conclusion, on account of the 10 animals been maintained in the same physical space and submitted to the same handling system, physiological variations, which are characteristic of this species, can be apointed as the reason of these differences.

Ovarian follicular and luteal dynamics in wapiti during seasonal transitions.

Transitions from the anovulatory to the ovulatory season (n=20) and ovulatory to anovulatory season (n=11), were monitored daily by transrectal ultrasonography in wapiti. In 17 of 20 observations, the first interovulatory interval (IOI) was short (9.1+/-0.3 days; mean+/-S.E.M.) compared with later in the ovulatory season (21.3+/-0.1) and the last IOI (21.2+/-0.6 days). With one exception, the short IOI were composed of only one wave of follicular development. Subsequent IOI were composed of two or three waves. Maximum diameters of the first two ovulatory follicles were similar (11.3+/-0.4 mm versus 11.3+/-0.2 mm), but both were larger (P<0.05) than the last two ovulatory follicles of the ovulatory season (10.3+/-0.3 and 10.1+/-0.4 mm). Multiple ovulations occurred in three hinds at the first ovulation of the season and in one hind at the second ovulation, but were not at any other time. Day-to-day profiles of CL diameter and plasma progesterone concentration were smaller (P<0.05) for short versus long IOI. Maximum diameter (12.8+/-0.6 mm versus 12.5+/-0.6 mm) and the diameter profile of the last CL of the season were not different from that of the previous CL. In summary, transition to regular ovulation occurred over a 3-week interval and was preceded by one short IOI (9 days). Multiple ovulations were detected only at the onset of the ovulatory season. The characteristics of the last IOI of the ovulatory season were similar to those reported during the rut. The wave pattern of follicle development was maintained throughout both fall and winter transition periods and follicular wave emergence was preceded by a surge in serum FSH concentrations. Transition to anovulation occurred over a 3-month interval and was marked by a failure of the dominant follicle to ovulate after a typical luteal phase.

Ovarian follicular and luteal dynamics in wapiti during the estrous cycle.

The reproductive tracts of 13 mature hinds were examined daily by transrectal ultrasonography and blood samples were taken daily from October to January to characterize follicular, luteal, and endocrine dynamics in wapiti during the estrous season. Follicle development occurred in waves characterized by regular, synchronous development of a group of follicles in temporal succession to a surge in serum FSH concentration. The mean interovulatory interval was 21.3 +/- 0.1 d, but was shorter in hinds exhibiting two follicular waves than in hinds exhibiting three and four waves (P < 0.05). The interwave interval was similar among waves in two-wave cycles and the first wave of three-wave cycles. All other interwave intervals in three- and four-wave cycles were shorter (P < 0.05). The maximum diameter of the dominant follicle of the first wave was similar among two-, three-, and four-wave cycles. For all other waves in three- and four-wave cycles, the maximum diameter was smaller (P < 0.05). Corpus luteum diameter and plasma progesterone concentrations were similar between two- and three-wave cycles, but the luteal phase was longer (P < 0.05) in four-wave cycles. The dominant follicle emerged at a diameter of 4 mm at 0.4 +/- 0.1 and 0.8 +/- 0.1 d before the largest and second largest subordinate follicles, respectively. The follicle destined to become dominant was larger (P < 0.05) than the largest subordinate follicle one day after emergence, which coincided with the first significant decrease in serum FSH concentration. We concluded that the estrous cycle in wapiti is characterized by two, three, or four waves of follicular development (each preceded by a surge in circulating FSH), that there is a positive relationship between the number of waves and the duration of the cycle, and an inverse relationship between the number of waves and the magnitude of follicular dominance (diameter and duration of the dominant follicle).

Acid-base, blood gas, and physiologic parameters during laparoscopy in the head-down position in white-tailed deer (Odocoileus virginianus).

The objective of this project was to evaluate the acid-base, blood gas, and physiologic parameters of white-tailed deer (Odocoileus virginianus) during laparoscopy in the head-down position. Eleven white-tailed does were captured and then immobilized with xylazine (6 mg/kg i.m.) and ketamine (7 mg/kg i.m.). The deer were intubated orotracheally and maintained with isoflurane in oxygen. The deer were positioned in dorsal recumbency and positive pressure ventilated. Heart rate (HR), arterial blood pressure, end-tidal carbon dioxide concentration (FE/CO2), and CO2 insufflation pressure were recorded every 5 min. Respiratory parameters, plasma electrolytes, and peak inspiratory pressure were measured immediately before tilting deer in the head-down position (45-55 degrees), 5 min after tilting, and immediately before the end of the procedure (while tilted). Butorphanol (0.05 mg/kg i.m.) was administered at the end of the procedure and yohimbine (0.2 mg/kg i.v.) administered before release. The deer weighed 52 kg (28-70 kg) [median (minimum-maximum)]. The peak inspiratory pressure in dorsal recumbency while still horizontal was 25 cm H2O (16-28 cm H2O), which increased to 29 cm H2O (18-46 cm H2O) after tilting (P = 0.02). PaO2, PaCO2, FE/CO2, and pH did not change after tilting in the head-down position or after insufflation. HR did not change during the anesthetic period. Mean arterial pressure did not change after tilting or abdominal insufflation, but decreased by the end of the anesthetic period (approximately 1 hr). Time from intubation to extubation was 117 min (72-170 min) (n = 5) and surgery time was 31 min (17-60 min; n = 10). We conclude that captured white-tailed deer have minimal derangements to acid-base, blood gas, or physiologic parameters during laparoscopy in the head-down position with abdominal insufflation, and thus this procedure may be performed safely in ventilated white-tailed deer.

Progesterone determination in Iberian red deer by time-resolved fluorometry: an alternative method to RIA.

The validation for Iberian red deer of a commercially available Time-resolved fluoroimmunoassay (TR-FIA) designed for analysis of progesterone in human beings was carried out. Intra-assay coefficients of variation ranged from 3.6% to 7.4%, while inter-assay coefficients of variation varied from 5.2% to 15.5%. Accuracy, evaluated by comparing results yielded by TR-FIA with those obtained from a validated radioimmunoassay (RIA) in the measurement of 14 samples, provided a high regression coefficient (R(2)= 0.93). Different progesterone concentrations added to pool plasma showed percentages of recovery that ranged between 102.6% and 82.48%. The limit of detection was 0.102 nmol/L. The results obtained indicate that the present method is suitable for the measurement of progesterone in female Iberian red deer.

Serum progesterone changes in luteal cyclicity and duration of estrous cycle in Formosan sika deer (Cervus nippon taiouanus) hinds.

A study was conducted to investigate the serum progesterone (SP(4)) profiles and duration of estrous cycles in the farmed Formosan sika deer (FSD; Cervus nippon taiouanus) during the major breeding season. Five parous, open and non-milking hinds were allotted to collect peripheral blood samples twice weekly for P(4) measurement by radioimmunoassay beginning at the initiation of the rutting season indicated by rutting behaviors of the sexually mature stags. The hinds were polyestrous as proved by cyclic changes of SP4 levels. After the presumptive estrus shown by the lowest concentration of SP(4) (0.20+/-0.01 ng/ml), this ovarian hormone markedly elevated on day 7 of the cycle (1.67+/-0.11 ng/ml), reached plateau (3.15+/-0.16 ng/ml, P<0.01) during days 11 to 18, and then declined to the basal levels in the subsequent estrus. It is concluded that mean duration of the estrous cycle in FSD during the major rutting season is 19.3 days with a range of 17 to 21 days, and that the patterns of circulating progesterone profiles during the estrous cycles of the FSD are similar to those of other deer species so far investigated.

Effects of GnRH agonist (leuprolide) on reproduction and behaviour in female wapiti (Cervus elaphus nelsoni).

Fertility control offers a potential alternative to traditional methods for regulating the growth of overabundant wild ungulate populations. However, current technology is limited due to practical treatment application, undesirable side-effects and economic considerations. A promising non-steroidal, non-immunological approach to contraception involves the use of a potent GnRH agonist. Two experiments were conducted to evaluate the effectiveness of a GnRH agonist (leuprolide) for controlling fertility in captive female wapiti and to assess physiological and behavioural side-effects of the treatment. In Expt 1, the optimum dose of agonist treatment was determined by measuring serum LH response of eight female wapiti to four formulations of leuprolide (0, 45, 90 and 180 mg) administered as a subcutaneous (s.c.) bioimplant. In Expt 2, the effects of leuprolide on wapiti pregnancy rates, duration of suppression of serum LH and progesterone secretion, and short-term behavioural and physiological side-effects were evaluated. All concentrations of leuprolide in Expt 1 were equally effective in reducing serum LH to non-detectable values throughout the 130 day trial. In Expt 2, leuprolide administered before the breeding season was 100% effective at preventing pregnancy in treated females. Serum LH and progesterone were reduced to baseline values by day 92 and remained at this concentration for 195-251 days after treatment, and returned to pretreatment concentrations in the following breeding season. Reproductive behaviour rates were similar for treated and untreated wapiti for all behaviour categories for both the breeding and post-breeding seasons. Haematology and blood chemistry parameters of treated and un-treated females were similar, and seasonal intake and body weight dynamics appeared normal. In conclusion, leuprolide is a safe, effective contraceptive agent and can potentially suppress fertility in female wapiti for one breeding season.

Fecal progesterone analysis by time-resolved fluoroimmunoassay (TR-FIA) for monitoring of luteal function in the sika doe (Cervus nippon centralis).

Fecal progesterone content was measured by time-resolved fluoroimmunoassay (TR-FIA) in the sika doe (Cervus nippon). The total recovery rate of fecal progesterone by twice extraction with diethylether was about 60%. The displacement curve of TR-FIA with serial doses of fecal extract (0.156-5.0 mg feces) was closely parallel to that of the reference standard. Fecal progesterone content was correlated with that of plasma (r=0.829, n=16), but the values were 100-fold higher in feces than in plasma. Fecal progesterone content periodically changed during the breeding season suggesting the estrous cycle in the doe. The fecal progesterone content was higher between the estruses, and decreased after estrus. The time between the onset of estrous signs and the lowest fecal progesterone content was 1-2 days suggesting the time required for hepatic metabolism and intestinal passage. Fecal progesterone content was also decreased around the time of vaginal discharge. The discharge took place within a few days, suggesting a short luteal phase. Not of all decreases in fecal progesterone values were preceded by estrous behavior or vaginal discharge. Fecal progesterone content was further increased in pregnancy rather than in the preceding estrous cycle and the levels were maintained up to term. These results suggest that fecal progesterone measurement is a useful tool for non-invasive analysis of luteal function in the sika doe. The TR-FIA kit, designed for the human hospital market, was shown to be successfully utilized for fecal assay in the sika doe with minor modifications.

Acid-base status and blood gas arterial values in free-ranging sika deer hinds immobilized with medetomidine and ketamine.

Ten free-ranging female sika deer (Cervus nippon) were captured to obtain the reference values for acid-base status and blood gas when immobilized with the combination of medetomidine and ketamine. The mean +/- SE of PaCO2, PaO2, and HCO3- were 58.1 +/- 6.1 mmHg, 58.8 +/- 6.4 mmHg, and 36.0 +/- 4.4 mmol/l, respectively. Although acidosis and alkalosis occurred in three and two animals, respectively, no serious conditions were observed. The blood values, however, suggest that some degree of hypoxemia and respiratory acidosis with metabolic alkalosis are developed. The trapped deer showed a significantly higher than normal rectal temperature reflective of exertion.