Cross-amplification tests of ungulate primers in the endangered Neotropical pampas deer (Ozotoceros bezoarticus)

In cross-species amplification tests of 15 ungulate primers in pampas deer, five were retained to form a small panel of highly polymorphic loci that could be used to efficiently screen populations of this endangered species. The polymerase chain reactions were performed incorporating the universal fluorescent labeled M13 (-21) primer. In 69 pampas deer, average allelic diversity was 15, expected heterozygosity was 0.869 and the mean polymorphic information content value was 0.847. Paternity exclusion probabilities over loci were NE-1P = 0.01336 and NE-2P = 0.00135, and combined non-exclusion probability of identity was P(ID) = 3 x 10(-8).     

Seasonal pattern of agonistic and marking behaviour of adult and young pampas deer (Ozotoceros bezoarticus) males

acta ethologica - Pampas deer (Ozotoceros bezoarticus) is an endangered species form the southern region of South America. The aim of this study was to describe and compare the seasonal changes in...     

Seasonal changes in fecal testosterone concentrations and their relationship to the reproductive behavior, antler cycle and grouping patterns in free-ranging male Pampas deer (Ozotoceros bezoarticus bezoarticus)

The purpose of this study was to validate noninvasive endocrine monitoring techniques for Pampas deer and to evaluate seasonal changes in testicular steroidogenic activity and their correlation to reproductive behavior, antler cycle and group size. Thus, fecal samples, behavioral data and observations of antler status were collected at monthly intervals during 1 year from free-ranging Pampas deer stags (three radio-collared individuals and 15 random individuals) living in Emas National Park, Brazil (18 degrees S latitude). Fecal steroids were extracted using 80% methanol and steroid concentrations were quantified by a commercial enzyme immunoassay (EIA). Fecal testosterone concentrations peaked in December-January (summer), March (early autumn) and in August-September (winter-spring), with minimal values from April-July. Reproductive behavior had two peaks, the first in December-January, characterized by predominately anogenital sniffing, flehmen, urine sniffing, chasing and mounting behavior, and the second peak in July-September (behavior primarily related to gland marking). There were significant correlations between fecal testosterone and reproductive behavior (r=0.490), and between fecal testosterone and antler phases (r=0.239). Antler casting and regrowth occurred under low testosterone concentrations, whereas velvet shedding was associated with high concentrations of testosterone. We inferred that Pampas deer stags exhibited a seasonal cycle that modulated sexual behavior and the antler cycle, and we concluded that fecal steroid analysis was a practical and reliable non-invasive method for the evaluation of the endocrine status of free-ranging Pampas deer.     

Discovery of an undescribed protostrongylid nematode from the endangered pampas deer (Ozotoceros bezoarticus celer) in Argentina

Dorsal-spined protostrongylid nematode larvae (Metastrongyloidea: Protostrongylidae) were recovered from the feces of the endangered pampas deer (Ozotocerus bezoarticus celer) in Campos del Tuyú Wildlife Reserve, Bahia Samborombón, Argentina. Partial DNA sequences from the large subunit ribosomal RNA (LSU rRNA) gene and from the second internal transcribed spacer region (ITS2) were amplified, cloned, sequenced, and compared to those of other nematodes. Nucleotide alignment and phylogenetic analysis of the sequences indicate that this protostrongylid nematode is most closely related to Parelaphostrongylus spp. as inferred from the LSU rRNA sequence analysis. Analysis of the ITS2 spacer indicated that the pampas deer protostrongylid is nested in a clade containing Parelaphostrongylus and Elaphostrongylus spp. These sequences differed considerably from those of other protostrongylid nematodes, and were most similar to those of Parelaphostrongylus spp. and Elaphostrongylus spp. in spite of clear variability from both genera. These results suggest that the protostrongylid from pampas deer is an undescribed nematode that likely belongs in the subfamily Elaphostrongylinae.     

Conservation genetics of the endangered Pampas deer (Ozotoceros bezoarticus)

The Pampas deer ( Ozotoceros bezoarticus L. 1758) is the most endangered neotropical cervid, and in the past occupied a wide range of open habitats including grassland, pampas, savanna, and cerrado (Brazil) from 5° to 41° S. To better understand the effect of habitat fragmentation on gene flow and genetic variation, and to uncover genetic units for conservation, we examined DNA sequences from the mitochondrial control region of 54 individuals from six localities distributed throughout the present geographical range of the Pampas deer. Our results suggest that the control region of the Pampas deer is one of the most polymorphic of any mammal. This remarkably high variability probably reflects large historic population sizes of millions of individuals in contrast to numbers of fewer than 80 000 today. Gene flow between populations is generally close to one migrant per generation and, with the exception of two populations from Argentina, all populations are significantly differentiated. The degree of gene flow was correlated with geographical distance between populations, a result consistent with limited dispersal being the primary determinant of genetic differentiation between populations. The molecular genetic results provide a mandate for habitat restoration and reintroduction of Pampas deer so that levels of genetic variation can be preserved and historic patterns of abundance can be reconstructed. However, the source of individuals for reintroduction generally should be from populations geographically closest to those now in danger of extinction.     

Effects of environmental conditions, human activity, reproduction, antler cycle and grouping on fecal glucocorticoids of free-ranging Pampas deer stags (Ozotoceros bezoarticus bezoarticus)

In this study, a commercial enzyme immunoassay (EIA) was validated in detecting glucocorticoids in Pampas deer feces, in order to investigate the influence of several factors on the adrenocortical function. Fecal samples, behavioral data and information concerning male grouping and antlers status were collected at a monthly basis during a 1 year period from free-ranging stags living at Emas National Park, Brazil (18 degrees S/52 degrees W). The results revealed that concentrations of fecal glucocorticoids in winter were significantly higher than those corresponding to spring and summer. In addition, dry season data presented higher levels than during the wet season. Significant difference was found between fecal levels of breeding stags in summer and nonbreeding stags, whereas no difference was observed between breeding stags in winter and nonbreeding stags. On the other hand, males from areas with frequent human disturbance exhibited higher glucocorticoid concentrations and flight distances than individuals from areas of lower human activity. Males with antlers in velvet had elevated levels compared with animals in hard antler or antler casting. Also, we found that glucocorticoid levels were higher in groups with three or more males than in groups with only one male. The flight distances showed positive correlation with fecal glucocorticoid. These data indicate that fecal glucocorticoid provides a useful approach in the evaluation of physiological effects of environment, inter-individuals relationship and human-induced stressors on free-ranging Pampas deer stags.     

Survey of Leptospira spp in pampas deer (Ozotoceros bezoarticus) in the Pantanal wetlands of the state of Mato Grosso do Sul, Brazil by serology and polymerase chain reaction

This work reports a survey of Leptospira spp in pampas deer (Ozotoceros bezoarticus) in the Pantanal wetlands of the state of Mato Grosso do Sul, Brazil by serology and polymerase chain reaction (PCR). Seventy pampas deer were captured in the dry season and surveyed using PCR, microscopic agglutination test (MAT) (n = 51) and by both techniques (n = 47). PCR detected infections in two pampas deer and MAT detected infections in three. Through sequencing and phylogenetic analyses, the PCR-amplified fragment detected in deer was identified as Leptospira interrogans. Serovars Pomona and Butembo were detected using MAT and the highest titre was 200 for serovar Pomona. Epidemiological aspects of the findings are discussed.     

Seasonal changes in plasma leptin concentration related to antler cycle in Iberian red deer stags

The aim of this study is to describe the leptin cycle in male Iberian red deer (Cervus elaphus hispanicus) and relate it to antler and testosterone cycles. An additional aim is to assess the relationship between the plasma leptin concentration during antlers’ growth and their final size. Therefore, blood from 21 Iberian red deer males was sampled monthly to analyse leptin and testosterone. At the same time the deer were weighed and their body condition was assessed. The length of antlers was measured every 2 weeks and, after casting, their final length and perimeters were taken. Leptin showed a seasonal cycle, with a peak in June that decreased as testosterone increased. Low values were observed in autumn, winter and early spring. The relationship observed between leptin and body mass or body condition score was different in spring, when plasma testosterone concentration is low, than in autumn, when testosterone increases. Leptin peak amplitude was positively related to final antler size. In conclusion, the relationship between leptin and body mass and body condition score changes through the year, possibly due to the influence of androgens and photoperiod. There was a positive relationship between plasma concentration of leptin during antler growth and final antler length.     

Seasonal variations in red deer (Cervus elaphus) hematology related to antler growth and biometrics measurements

The aim of the study was to relate seasonal hematology changes with the rest of physiological variations suffered by red deer, such as antler and biometrics cycle, and to assess the relationship between hematology and the effort performed in antler development. Blood samples were taken from 21 male red deer every 4 weeks during 18 months. Samples were analyzed for the main hematological parameters. Simultaneously, biometrics measurements were taken, such as antler length, body weight, body condition score, testicular diameter (TD), and thoracic and neck girth. All the blood cell types (erythrocytes, leukocytes, and platelets) showed seasonal variations, increasing as antler cleaning approached, as did hematocrit and hemoglobin. The final size of antlers was negatively related to leukocyte count, nonlymphoid leukocyte count, red cell distribution width, mean corpuscular hemoglobin, mean platelet volume, and TD, whereas it was positively related to body condition during antler growth. Huge seasonal variations in some hematological values have been found to be related to changes in antler and biometrics measurements. Since these variations are even greater than the caused by deer handling, they should be taken into account when evaluating hematology in deer populations.     

Seasonal variation of LH and testosterone in the smallest deer,the pudu (Pudu puda molina) and its relationship to the antler cycle

• 1.1. In order to obtain a seasonal profile of LH, three adult male pudu (Pudu puda, Molina) were sampled monthly from the saphenous vein for a period of one year.
• 2.2. A significant circannual variation of plasma LH levels was detected with an average peak value (1.77 ng/ml) recorded in February and nadir concentrations (0.19 ng/ml) observed in November.
• 3.3. The peak level of testosterone (1.54 ng/ml) was detected in March, the time of the rut.

     

Seasonal pattern of luteinizing hormone and testosterone pulsatile secretion in young adult red deer stags (Cervus elaphus) and its association with the antler cycle.

Blood from stages aged 15 months (n = 6) was sampled at monthly intervals every 30 min for 24 h for 12 months, at 45 degrees S in New Zealand. Three extra samplings each for 24 h were carried out at about the anticipated time of antler casting. All samples were analysed for luteinizing hormone (LH) and testosterone and the resulting data further analysed by the Pulsar pulse detection routine. The animals were kept indoors under natural daylength and were fed ad libitum. All animals were weighed, antler status and size recorded and testes diameter was measured on each sampling day. Mean LH and testosterone pulsatily and plasma concentration varied seasonally. LH pulse frequency was low during autumn (2.5 pulses in 24 h), winter (1.0-1.5 pulses in 24 h) and early spring (1 pulse in 24 h) and lowest in late spring (0.2 pulse in 24 h) before rising in summer (1.0-4.0 pulses in 24 h). LH pulse amplitude and mean plasma concentration were low (< 1 ng ml-1) from March to November (autumn-spring); both rose to a peak in January (summer) of 3.4 and 1.6 ng ml-1, respectively. Testosterone pulse frequency was generally similar to LH except that slightly more pulses of testosterone than of LH were detected from March to November and more pulses of LH from November to February (summer). Testosterone pulse amplitude fell from March to November (5.3 ng ml-1 to undetectable) although there was a conspicuous peak in July (midwinter) of almost 5 ng ml-1.(ABSTRACT TRUNCATED AT 250 WORDS)     

Morphological observation of antler regeneration in red deer (Cervus elaphus)

Deer antler offers a unique opportunity to explore how nature solves the problem of mammalian appendage regeneration. Annual antler renewal is an example of epimorphic regeneration, which is known to take place through initial blastema formation. Detailed examination of the early process of antler regeneration, however, has thus far been lacking. Therefore, we conducted morphological observations on antler regeneration from naturally cast and artificially created pedicle/antler stumps. On the naturally cast pedicle stumps, early antler regeneration underwent four distinguishable stages (with the Chinese equivalent names): casting of previous hard antlers (oil lamp bowl), early wound healing (tiger eye), late wound healing and early regeneration (millstone), and formation of main beam and brown tine (small saddle). Overall, no cone-shaped regenerate, a common feature to blastema-based regeneration, was observed. Taken together with the examination on the sagittal plane of each regenerating stage sample, we found that there are considerable overlaps between late-stage wound healing and the establishment of posterior and anterior growth centers. Observation of antler regeneration from the artificially created stumps showed that the regeneration potential of antler remnants was significantly reduced compared with that of pedicle tissue. Interestingly, the distal portion of a pedicle stump had greater regeneration potential than the proximal region, although this differential potential may not be constitutive, but rather caused by whether or not pedicle antlerogenic tissue becomes closely associated with the enveloping skin at the cut plane. Antler formation could take place from the distal peripheral tissues of an antler/pedicle stump, without the obvious participation of the entire central bony portion. Overall, our morphological results do not support the notion that antler regeneration takes place through the initial formation of a blastema; rather, it may be a stem cell-based process.     

The effect of cyproterone acetate on the antler cycle in red deer (Cervus elaphus L.)

The aim of the study was to test the effect of antiandrogen, cyproterone acetate (CA) on the antler cycle in the red deer (Cervus elaphus). CA was administered to three adult red deer stags (Edward, Fuks and Gacek) in weekly intervals. Edward and Fuks were given 600 mg + 600 mg of CA, whereas Gacek was given 600 mg + 300 mg. CA was injected during the hard antler phase: in mid-October (Edward), at the end of November (Fuks) and at the end of January (Gacek). CA caused the antler casting 17 to 22 days after the first injection. In all stags, the casting of antlers was followed by a period of intensive growth of new antlers. Edward was given CA at the end of October. This treatment was responsible for occurrence of the two antler cycles in the year of the experiment. When CA was administered during the middle of the hard antler phase an additional short antler cycle occurs followed by new antler growth. CA treatment in the later part of hard antler phase may cause a prolonged antler cycle.     

Allozymes and the genetics of antler development in red deer (Cervus elsphus)

In order to examine a previously hypothesized influence of selective hunting on allele frequency changes at some regularly polymorphic allozyme loci in red deer ( Cervus eluphus ), antler characters, serving as criteria for culling, were examined in relation to electrophoretic variation in two free-ranging populations of the Vosges, Eastern France, and an enclosure in Central France. When homozygous for the allele Idh-2 ¹²⁵, stags ≥ 2 years old had a significantly higher number of antler points (NAP). When homozygous for the allele Acp-2 ¹⁰⁰, stags older than 5 years had antlers that were significantly larger for a number of traits (NAP, main beam length and circumference, coronet circumference, brow tine length). Among younger stags, all antler traits in Acp-2 ¹⁰⁰ homozygotes were significantly smaller than in carriers of the alternative allele, Acp-2 ⁸⁵. Our data suggest the presence of at least two independent genetic components (one associated with early development of a high NAP, the other with generally large antler size in adults), affecting antler expression in red deer. Those genetic components, possibly major genes which are chromosomally linked with the allozyme loci studied, compensate or reinforce each other in their phenotypic effects. By playing a role in balancing benefits and costs of male reproductive success, they may be part of a genetic mechanism enabling the rapid adaptation of a population to various environmental and demographic conditions. The three populations studied originate from one another, and, based on an assessment of effective population sizes, it could be demonstrated that selective hunting for antler shape has changed allelic frequencies at the associated marker loci within a few generations.     

Effect of antler growth period on the chemical composition of velvet antler in sika deer (Cervus nippon)

The aim of this study was to provide basic information to allow improved scientific assessment of velvet antlers’ quality by investigating the change of chemical composition depending on antler growth period in sika deer (Cervus nippon). Twelve antlers harvested from sika deer stags (aged 4–5 years) by antler growth periods of 40 days (FDG) and 60 days (SDG) after the casting of antler buttons from the previous set were analysed to compare the chemical composition, such as crude protein, ash, ether extract, amino acid, collagen, glycosaminoglycans (GAGs), uronic acid and sialic acid. The weight of velvet antler in FDG was lower than that of the SDG (P<0.05). SDG had a higher (P<0.05) content of crude ash than FDG. FDG had a higher (P<0.05) content of crude protein than SDG. Amino acid composition was also higher in FDG than in SDG for all sections. The content of collagen was higher in SDG than in FDG for the middle and base (P<0.05) sections. However, collagen levels were exceptionally higher (P<0.05) in FDG than in SDG for the upper section. While the content of collagen was significantly higher (P<0.05) for the upper section than for the middle and base sections in FDG, this was significantly increased (P<0.05) downward from the upper to the base section in SDG. Uronic acid content was higher in FDG than SDG for all three sections but there was no significant difference between groups in the middle and base sections. The content of GAGs was significantly higher (P<0.05) in FDG than SDG for all three sections. The content of sialic acid was the same as that of GAGs. Consequently, in this study, velvet antler production was increased with the extension of antler growth period, but the contents of protein, total amino acid composition, GAGs, uronic acid and sialic acid were decreased and those of ash and collagen were increased. Therefore, it is expected that the quality of velvet antler may be decreased greatly by extension of antler growth period.     

Sika deer (Cervus nippon) velvet antler extract attenuates prostate cancer in xenograft model

The present study determines whether antler extract (AE) possesses inhibitory effects in a prostate cancer (PC) xenograft model and explores the underlying mechanism. After therapeutic intervention for two weeks, AE significantly inhibited prostate cancer xenograft tumor growth by 65.08%, and prostate-specific antigen (PSA) and serum dihydrotestosterone (DHT) levels. However, AE increased the serum testosterone level compared to the vehicle control group. Furthermore, our investigation of the inhibitory effects on angiogenesis and epithelial-to-mesenchymal transition (EMT)-related genes revealed that AE downregulated matrix metalloproteinase 2 (MMP)-2, (MMP)-9, vascular endothelial growth factor (VEGF), zinc finger protein (SNAIL1), twist-related protein 1 (TWIST1), and zinc-finger E-box-binding homeobox 1 (ZEB1) in vivo. In contrast, AE increased tissue inhibitor of MMP (TIMP)-1, (TIMP)-2, and E-cadherin. The results suggest that AE possesses potent anti-PC activity, and this is the first report on the anti-PC effect of AE in vivo.