Intrauterine insemination of farmed fallow deer (Dama dama) with frozen-thawed semen via laparoscopy

Estrus and ovulation of mature fallow does (n = 155) on two North American farms were synchronized by intravaginal silastic devices containing 0.3 g progesterone (CIDR-type G) for 14 d. Each of 151 does received laparoscopic intrauterine inseminations of either 50 x 10(6) (n = 125) or 25 x 10(6) (n = 26) frozen-thawed spermatozoa, 65 to 68 h after CIDR device withdrawal. Four does received intrauterine inseminations per vaginam of 50 x 10(6) spermatozoa 68 to 69 hours after CIDR device withdrawal. Semen from crossbred Dama dama dama x Dama dama mesopotamica sires was collected in New Zealand by electroejaculation. The overall pregnancy rate to artificial insemination, as assessed by rectal ultrasonography at Day 45, was 67.7%. The pregnancy rates for does receiving laparoscopic inseminations were 58.2% (Texas; 50 x 10(6) spermatozoa; n = 79 does); 80.8% (Texas; 25 x 10(6) spermatozoa; n = 26 does) and 76.1% (New York; 50 x 10(6) spermatozoa; n = 46 does). Three of the four does receiving intrauterine inseminations per vaginam became pregnant to the frozen-thawed semen.     

Embryo transfer in fallow deer (Dama dama): superovulation, embryo recovery and laparoscopic transfer of fresh and cryopreserved embryos

Multiple ovulation-embryo transfer (MOET) protocols for farmed fallow deer (Dama dama ) were investigated in a series of 3 experiments. A total of 37 donors, of either European (D.d. dama ; n = 30) or Mesopotamian hybrid (D.d. mesopotamica x D.d. dama ; n =7) genotype, each received an intravaginal silastic device containing 0.3 g progesterone (CIDR((R))-type G device) for 14 d and injections of 0.5 units ovine FSH (8 x 0.06 unit injections from Days 10 to 14 of device insertion) and 100 IU PMSG (either with the first or last FSH injection). All donors received laparoscopic intrauterine inseminations of fresh semen (50 x 10(6) spermatozoa) from a Mesopotamian sire 36 h after withdrawal of CIDR((R)) devices. Embryos were recovered by laparotomy on Day 6 (Day 0 = estrus). Mean ovulation rates for the 3 experiments were 8.1, 9.8 and 7.0, with no effect of PMSG timing (P>0.10). However, embryo recovery rates, albeit low throughout the study (29.6%), were significantly improved with later PMSG administration (33.9 vs 20.1%; P<0.05). Hybrid and European donors performed in a similar manner. A range of embryo development stages was recovered throughout the study. In 2 experiments laparoscopic transfer of embryos to 48 recipient does treated previously with intravaginal CIDR((R)) devices for 14 d yielded a total pregnancy rate of 37.5%. In the experiment with fresh embryos, the use of clenbuterol to reduce uterine turgidity resulted in a higher proportion of does conceiving (3 4 ; 75%) compared with that of the untreated does (0 6 , 0%; P<0.05). In the second experiment, in which all the does routinely received clenbuterol, 10 19 (53%) and 5 19 (26%) does conceived following the transfer of fresh and cryopreserved embryos, respectively (P<0.05). While the overall efficiency of the MOET program was low (equivalent of 0.9 to 1.0 surrogate pregnancies per donor), improvements in the recovery rate of transferable embryos have considerable potential for genetic improvement of farm stock and captive propagation of endangered Mesopotamian fallow deer through maternal surrogacy programs.     

Use of pregnancy-specific protein-B and estrone sulfate for determination of pregnancy on Day 49 in fallow deer (Dama dama )

The objective of this study was to determine if pregnancy specific protein-B (PSPB) and estrone sulfate (E(1)SO(4)) could be used to determine pregnancy status in fallow deer (Dama dama ). Forty mature does were synchronized for estrus with an intravaginal progesterone-releasing device (CIDR) and then artificially inseminated via laparoscopy with frozen semen on one day. Ultrasound examination and jugular blood sampling were done 49 days later. Transrectal ultrasonography was done to presumptively determine the pregnancy status at the time of blood sampling. Serum estrone sulfate concentrations were significantly (P < 0.05) greater in pregnant (n=31) than nonpregnant (n=9) females at 49 days of gestation (166.7 +/- 25.9 pg/ml vs 36.3 +/- 11.1 pg/ml, respectively). The percentage of [(125)I]PSPB bound was significantly (P < 0.01) lower when sera of pregnant (n=29) versus nonpregnant (n=9) females was added to RIA tubes (63.7 +/- 1.6% vs 98.1 +/- 1.6%, respectively). There were 30 fawns born from the group of females that were diagnosed pregnant based on ultrasound. We conclude that estrone sulfate and PSPB can be used to determine pregnancy status in fallow deer at 49 days of gestation.     

A method of artificial insemination in captive White-tailed deer (Odocoileus virginianus )

Production of fawns by artificial insemination in captive White-tailed deer (Odocoileus virginianus ) has been accomplished by using frozen-thawed spermatozoa. The purpose of this study was to determine if frozen-thawed semen deposited at the posterior face of the os cervix could produce conception. Five hand-raised female White-tailed deer and one hand-raised male White-tailed deer were used over two breeding seasons 1984-1985 and 1985-1986. The vasectomized buck was ued to detect estrus in the does. The does were inseminated with frozen-thawed semen containing at least 100 million live normal cells with a 60% or higher motility. The artificial insemination catheters used in this study worked well, but due to the small size of the cervix, the catheter could only be passed up to the first cervical ring, the site at which the semen was deposited. Over two breeding seasons, nine does were inseminated with frozen-thawed spermatozoa; each doe was inseminated once each estrous cycle at one of the following times: 0, 6, 12, 18, 24 or 30 h. after detection of estrus. Of the nine does inseminated with frozen-thawed spermatozoa, six conceived and carried to term 11 healthy normal fawns, yielding an overall conception rate of 67%.     

Light and scanning electron microscopy of fallow deer (Dama dama) spermatozoa

No basic differences in size (mean +/- s.d. for at least 300 spermatozoa), shape and ultrastructure of the spermatozoa of fallow deer were detected (1) in comparison to other artiodactyls, (2) between different fallow bucks, and (3) between different months of the fertile season. The total length of the normal spermatozoon was 67.2 +/- 1.2 microns. The flat, paddle-shaped head was 8.2 +/- 0.3 microns long, 4.4 +/- 0.2 microns for the greatest width, 1.9 +/- 0.2 microns for basal width and, approximately 0.7 microns in thickness. The tail measurements were 13.7 +/- 0.3 microns for the midpiece, 0.5 +/- 0.1 microns for the diameter of the midpiece, 42.6 +/- 0.9 microns for the principal piece, and 2.7 +/- 0.6 microns for the endpiece. Spermatozoa with abnormalities such as cytoplasmic remnants and droplets, bent and coiled tails, as well as microcephalic forms were observed.     

Early pregnancy detection and the hormonal characterization of embryonic-fetal mortality in fallow deer (Dama dama)

The objectives of this investigation were to 1) determine serum concentrations of progesterone (P4), estrone sulfate (E1S) and pregnancy-specific protein B (PSPB) from estrus synchronization through mid-gestation in the fallow doe (Dama dama) and 2) characterize the hormonal profiles of does whose embryos or fetuses died in utero. Ten fallow does were synchronized for 14 d with an intravaginal P4-releasing device (CIDR) and were naturally mated after CIDR removal. Blood samples were collected at CIDR insertion, CIDR removal and at intervals through Day 203 post-CIDR removal for analysis of P4, E1S and PSPB by radioimmunoassay (RIA). Ultrasonography was performed on Days 49 and 69 post-CIDR removal. Serum P4 at the time of CIDR insertion was 4.8 +/- 0.6 ng/ml, and at CIDR withdrawal it was 6.2 +/- 0.3 ng/ml. Concentrations of E1S and PSPB were nondetectable at CIDR insertion. Serum E1S was highest at Day 93, and PSPB was first detectable in pregnant does at Days 27 to 30 post-CIDR withdrawal. Ultrasonography on Day 49 revealed that 6 does were pregnant, 2 were not pregnant and 2 others were diagnosed originally as early pregnant. At Day 69, ultrasonography revealed that 6 does (60%) were pregnant and 4 (40%) were not. A comparison of the ultrasonographic and hormonal data indicated that the 2 does diagnosed as early pregnant on Day 49 had conceived but had lost the pregnancy. A third doe which was pregnant on Day 69 lost the fetus later in gestation. Hormonal profiles of does whose embryo or fetus had died were characterized by erratic P4 and E1S profiles, with PSPB becoming undetectable in the 3 does by Days 49, 65 and 80 post-CIDR removal. These data 1) demonstrate the timing for the collection of serum samples for determining early pregnancy in fallow does using 3 hormonal methods and 2) characterize the hormonal profiles of 3 fallow does with embryonic-fetal loss.     

The collection and examination of semen of the Reindeer (Rangifer tarandus)

Semen was collected from all the mature Reindeer bulls (9) in the Glenmore herd, both normal and vasectomized. Nineteen samples of semen were obtained. Reindeer semen is a viscous fluid, generally amber in colour: after vasectomy its viscosity is much lower. The mean volume of the ejaculates was 0–5 ml. and mean sperm density 467 × 10⁴ spermatozoa. Motility was low in comparison with other mammals and the proportion of eosinophilic spermatozoa was high (74.5%). Other determinations included measurements of the size of unstained spermatozoa, and of fructose and citric acid concentration in the semen.     

Phylogeography of the last surviving populations of Rhodian and Anatolian fallow deer (Dama dama dama L., 1758)

The European fallow deer ( Dama dama dama ) is one of the most widespread cervids, and its distribution has been heavily affected by man. At present, only one wild autochthonous population is reputed to survive in Anatolia, but its census size is dramatically decreasing. This means that a significant portion of the ancestral genetic diversity of this taxon is seriously threatened. In the present study, a portion of the mitochondrial DNA (D-loop) in 37 D. d. dama specimens from three Mediterranean sites, Turkey, the island of Rhodes (Greece), and the Italian Peninsula, and seven individuals of the Persian fallow deer, Dama dama mesopotamica , was sequenced, and the results from the data analysis are interpreted in light of current archaeozoological and biogeographical knowledge. We conclude that: (1) D. d. mesopotamica is strongly differentiated from D. d. dama , confirming the results of previous genetic studies and (2) the Rhodian populations of D. d. dama , founded by humans in Neolithic times, possess a set of mitochondrial lineages, found in no other study populations. The persistence of these haplotypes is particularly significant because human-mediated processes (e.g. domestication) usually result in genetic depletion and erosion of an ancestral genetic pool. In the case of the Rhodes' population of fallow deer, we hypothesize that, during the foundation of this population, humans unknowingly preserved a remarkable portion of the original genetic diversity of the source population(s).  © 2008 The Linnean Society of London, Biological Journal of the Linnean Society , 2008, 93 , 835–844.     

Experimental tests of copying and mate choice in fallow deer (Dama dama)

In fallow deer (Dama dama), as well as in other lek-breedingungulates, receptive females arriving at leks commonly joinmales that are defending large harems. This tendency enhancesdifferences in harem size and mating success between males.It could occur because females independendy move to the samemales, because females are attracted to males with females,or because females are attracted to each other. Using controlledexperiments with estrous female fallow deer, we show that, althoughfemales are more attracted to males with harems than to thosewithout, they are as frequently attracted to groups of femaleswithout a male as to female groups with males. We conclude thatfemale fallow deer joining leks are attracted to each otherand copy each other's movements. As yet, there is no firm evidencein fallow deer or in other lek-breeding ungulates that femalescopy each other's choice of mating partners. Key words: Damadama, fallow deer, lek breeding, mate choice, copying behavior.[Behav Ecol 4: 191–193 (1993)]     

Inter-specific competition from fallow deer Dama dama reduces habitat quality for the Italian roe deer Capreolus capreolus italicus

The roe deer of Mediterranean habitats in the central and southern parts of Italy has recently been recognised as a distinct subspecies, Capreolus capreolus italicus . A population of this endangered subspecies has been monitored in the Preserve of Castelporziano, near Rome, since 1988. We observed an abrupt population decline in 2000, which may severely threaten the sustainability of this population. We evaluated the hypothesis that competition by fallow deer may be a principal cause of this decline. By a new and innovative methodology, we modelled the spatial distribution of fallow deer density (FDD) in the study area to show that 1) habitat quality for roe deer was an inverse function of FDD, 2) habitat apportionment between fallow and roe deer increased as a function of FDD and – by applying structural equation modelling – 3) FDD was superior to habitat composition in explaining observed variations in home range size and probably in habitat quality for roe deer. This analysis is the first to document that inter-specific competition may influence the spatial behaviour of a deer species leading to poor phenotypic performance in the inferior competitor. We conclude that the conservation of this relict population would benefit by reducing fallow deer numbers at Castelporziano and from other measures aimed to decrease the level of inter-specific competition.     

Rumen Ciliate Protozoa in Ohio White-tailed Deer (Odocoileus virginianus)

ABSTRACT. Rumen contents were obtained from 23 white-tailed deer (Odocoileus virginianus), located in the eastern portion of central Ohio. Samples were taken during late fall and winter over a 4-yr period, 1986–1990. Protozoan numbers ranged from 0.002 to 7.25 × 10⁶ per ml of rumen contents, with a mean of 2.96 × 10⁶. Six deer had protozoan concentrations higher than any values previously reported in ruminants. In all 23 animals, Entodinium dubardi was the only ciliate protozoan species present.     

Molecular identification of two Sarcocystis species in fallow deer (Dama dama) from Lithuania

Due to the lack of molecular research conducted, little is known about Sarcocystis species diversity in the fallow deer (Dama dama). Until now, Sarcocystis jorrini and Sarcocystis morae were described to form sarcocysts in the muscles of this host. In the present study diaphragm muscle samples of free-ranging fallow deer from Lithuania were investigated for Sarcocystis species. Sarcocysts were detected in 39 out of 48 (81.3%) fallow deer examined. Under a light microscope two types of sarcocysts having hair-like and finger-like protrusions were observed. Based on DNA sequence analysis of cox1 and 18S rDNA, two species, S. morae and Sarcocystis entzerothi were identified. In prior studies, the latter species was only detected in Lithuanian roe deer (Capreolus capreolus) and in sika deer (Cervus nippon). The haplotype network of S. morae sequences specified close relationships between haplotypes found in the same country. According to current knowledge, the fallow deer is characterised by low Sarcocystis species richness as compared with other cervid species from Europe.     

Context-related acoustic variation in male fallow deer (Dama dama) groans

While social and behavioural contexts are known to affect the acoustic structure of vocal signals in several mammal species, few studies have investigated context-related acoustic variation during inter-sexual advertisement and/or intra-sexual competition. Here we recorded male fallow deer groans during the breeding season and investigated how key acoustic parameters (fundamental frequency and formant frequencies) vary as a function of the social context in which they are produced. We found that in the presence of females, male fallow deer produced groans with higher mean fundamental frequency when vocal males were also present than they did when no vocal males were in close vicinity. We attribute this to the increased arousal state typically associated with this context. In addition, groan minimum formant frequency spacing was slightly, but significantly lower (indicating marginally more extended vocal tracts) when males were alone than when potential mates and/or competitors were nearby. This indicates that, contrary to our predictions, male fallow deer do not exaggerate the acoustic impression of their body size by further lowering their formant frequencies in the presence of potential mating partners and competitors. Furthermore, since the magnitude of the variation in groan minimum formant frequency spacing remains small compared to documented inter-individual differences, our findings are consistent with the hypothesis that formants are reliable static cues to body size during intra- and inter-sexual advertisement that do not concurrently encode dynamic motivation-related information.     

Successful uterine insemination of fallow deer with fresh and frozen semen

Six fallow does were inseminated directly into the uterine horns 72 h (three does) or 78 h (three does) after the removal of progestagen intravaginal sponges. Three does were inseminated with fresh (two at 72 h and one at 78 h) or frozen-thawed (one at 72 h and two at 78 h) semen. The semen used had been collected by electroejaculation and had been stored for 2 yr in liquid nitrogen in a Tris, citric acid, glycerol diluent containing 2.25% egg yolk. Three does each produced a live fawn to insemination and all does had been inseminated 72 h after removal of sponges; two with fresh semen and one with frozen semen. The remaining three does failed to conceive to insemination, but did produce fawns to mating at a subsequent estrus.     

Suckling time and fawn growth in fallow deer (Dama dama)

The total suckling time and the fawns' absolute increase in weight from birth to weaning were studied in a group of fallow deer over two consecutive calving seasons. The aim was to clarify the use of suckling time as a predictor of parental input in fallow deer. When all subjects from both years are taken into account, no positive relation between suckling time and growth was found. This was due to at least two important factors: (i) fawns to primiparous mothers sucked relatively longer, with no corresponding increase in weight, compared to multiparous mothers; and (ii) suckling time during the first two weeks after birth seems to be an inadequate measure of milk transfer.     

Foot infections associated with Arcanobacterium pyogenes in free-living fallow deer (Dama dama)

We describe foot infection associated with Arcanobacterium pyogenes in three adult male free-living fallow deer (Dama dama) from Sueve Regional Hunting Reserve (Principality of Asturias, Spain). Affected fallow deer were culled in November 1997 and 1998 during the hunting season. Necropsy, radiography, and microbiologic analysis were carried out for each animal. Unilateral swelling of one extremity at the coronary band was observed in all three cases. Areas of bone loss, severe periosteal reaction, and soft tissue swelling were seen on radiography. Lead fragments were observed in one fallow deer. Seven bacterial species were isolated, but only Arcanobacterium pyogenes was routinely found. Weather conditions in the area (mild temperatures and high humidity), the land (alternating pasture land and rock), the animal population density (both fallow deer and domestic herds of cows, horses, sheep, and goats, live side by side in the same areas), and hunting activities could be related to the frequency of these infections.     

Anaplasma phagocytophilum infection in a fallow deer (Dama dama) population in a preserve of central Italy

From autumn 2004 to spring 2005, 70 fallow deer (Dama dama), 27 female and 43 male, living in a natural preserve of central Italy were examined by indirect immunofluorescence assay (IFA) to detect specific antibodies to Anaplasma phagocytophilum. Thirty-one (44.28%) sera scored positive: in particular 10 fallow deer (8 male and 2 female) scored positive at 40 antibody titer, 21 deer (8 male and 13 female) at > or = 80 titer. EDTA anticoagulated blood samples collected from 29 of the 70 deer examined were tested by a nested-PCR assay to disclose a 546 bp fragment, specific of A. phagocytophilum 16S rRNA gene. Twenty (72.41%) blood samples (8 male and 13 female deer) resulted positive. Fifteen PCR-positive deer also resulted positive to IFA, whereas the remaining six did not show specific antibodies. Three serologically positive animals gave negative results at the nested PCR. Five deer scored negative both to serological tests and PCR.     

Identification of pregnancy-associated glycoproteins and alpha-fetoprotein in fallow deer (Dama dama) placenta

Background

This paper describes the isolation and characterization of pregnancy-associated glycoproteins (PAG) from fetal cotyledonary tissue (FCT) and maternal caruncular tissue (MCT) collected from fallow deer (Dama dama) pregnant females. Proteins issued from FCT and MCT were submitted to affinity chromatographies by using Vicia villosa agarose (VVA) or anti-bovine PAG-2 (R#438) coupled to Sepharose 4B gel. Finally, they were characterized by SDS-PAGE and N-terminal microsequencing.

Results

Four distinct fallow deer PAG (fdPAG) sequences were identified and submitted to Swiss-Prot database. Comparison of fdPAG with PAG sequences identified in other ruminant species exhibited 64 to 83% identity. Additionally, alpha-fetoprotein was identified in fetal and maternal tissues.

Conclusion

Our results demonstrate the efficacy of VVA and bovine PAG-2 affinity chromatographies for the isolation of PAG molecules expressed in deer placenta. This is the first report giving four specific amino acid sequences of PAG isolated from feto-maternal junction (FCT and MCT) in the Cervidae family.     

Effects of immunocontraception on behavior in fallow deer (Dama dama)

Immunocontraceptives can control growth in wild and captive populations; however, in doing so, they should not disrupt species‐typical behavior patterns. The presence of treated females could disrupt social interactions in a population; yet, few studies have examined effects of immunocontraception on behavior. The goal of this project was to determine whether behavior in a fallow deer (Dama dama) population vaccinated with porcine anti‐zona pellucida (pZP)‐immunocontraceptives differed from behavior in a population of unvaccinated deer. We predicted that pZP‐treated females would spend a higher percentage of time exhibiting mating and dominance behavior than nontreated females, and that males interacting with pZP‐treated females would spend a higher percentage of time demonstrating mating and aggressive behavior than males interacting with nontreated females. We recorded activity budgets of males and females in two fallow deer populations, one pZP‐treated and the other untreated, before, during, and after rut. Females did not differ in time spent displaying mating or dominance behavior compared to nontreated females. Males coexisting with nontreated females spent more time exhibiting aggressive behavior during rut than males living with pZP‐treated females, but males did not differ in time spent in mating behavior. Thus, immunocontraception did not seem to affect behavior adversely. However, sample sizes, living conditions, and sex ratios may have affected the results. Given these limitations, future research is needed to confirm our findings. Zoo Biol 27:49–61, 2008. © 2007 Wiley‐Liss, Inc.     

Mortality of fallow deer (Dama dama) experimentally-infected with meningeal worm, Parelaphostrongylus tenuis.

Six fallow deer (Dama dama) fawns died after receiving 25 to 150 infective larvae of Parelaphostrongylus tenuis. Fawns given higher doses usually died sooner (6 to 23 days) than those given lower doses (54 to 67 days). Early deaths were associated with severe acute peritonitis resulting from perforation of the intestinal wall; later deaths were associated with paralysis and inability to rise. Numerous adult P. tenuis were found within neural tissues of the brain and spinal cord in the three fawns with paralysis. One white-tailed deer (Odocoileus virginianus) exposed to infective larvae from the same source survived infection without exhibiting clinical signs and began passing larvae in feces 88 days post-exposure. At the doses used in this study, meningeal worm caused fatal infections in fallow deer. Results are compared to published observations of fallow deer naturally-infected with P. tenuis.