First description of gastrointestinal nematodes of Barbary sheep (Ammotragus lervia): the case of Camelostrongylus mentulatus as a paradigm of phylogenic and specific relationship between the parasite and its ancient host.

The gastrointestinal helminth fauna of 24 Barbary sheep or Aoudad (Ammotragus lervia sahariensis) maintained in the Parque de Rescate de la Fauna Sahariana (PRFS, CSIC, Almeria, Spain) was analyzed. Most animals (87.5 %) were parasitized, and multiple infections were highly present. The following species were identified: Camelostrongylus mentulatus, Teladorsagia circumcincta, Marshallagia marshalli, Ostertagia ostertagi, O. leptospicularis, O. lyrata, Haemonchus contortus, Teladorsagia trifurcata, Trichostrongylus vitrinus, T. colubriformis, T. probolorus, T. capricola, Nematodirus spathiger, N. abnormalis, N. filicollis, N. helvetianus, Trichuris spp. and Skrjabinema ovis. Teladorsagia circumcincta was the most prevalent nematode in abomasum (52.6 %) followed by C. mentulatus (50 %). However, this latter nematode had the greater mean intensity and abundance. In the small intestine, T. colubriformis and T. vitrinus had the highest prevalence (36.4 %); the last one showed also the greater mean intensity and abundance. It should be emphasized the presence of Skrjabinema ovis (prevalence 39.1 %) in the large intestine, showing the greater mean abundance and intensity, although with a low values. Camelostrongylus mentulatus could be the most primitive nematode of the family trichostrongylidae recovered in this study; attending to its high prevalence, mean abundance and mean intensity, the possible specificity between this parasite and the Aoudad is discussed.

Use of a neuroleptic in assisted reproduction of the critically endangered Mohor gazelle (Gazella dama mhorr).

Stress is a limiting factor in assisted reproduction in wild animals maintained in captivity and measures to reduce it should improve reproductive success. The effect of the long-acting neuroleptic (LAN) perphenazine enanthate was assessed on ovarian stimulation for the recovery of immature oocytes from Mohor gazelle (Gazella dama mhorr) and their subsequent in vitro maturation, fertilization and embryo culture. The viability of embryos after transfer was also examined. Perphenazine enanthate decreased activity levels and facilitated handling of treated animals when compared to controls. LAN-treated animals showed a more regular pattern of respiratory and heart rates and body temperature than controls; no major differences were found in hematological and biochemical parameters between groups. Perphenazine-treated females had lower plasma cortisol levels during the days of intense handling. No significant differences were found in the number of punctured follicles and recovered oocytes between groups. The percentage of mature oocytes per female was significantly higher in the LAN-group. Fertilization and cleavage rates were not significantly different between groups. Embryos developed in culture but none reached the blastocyst stage, and those transferred to the oviduct of synchronized recipients did not develop to term. In conclusion, treatment of females with perphenazine enanthate during ovarian stimulation did not have negative effects on maturation, fertilization and embryo development in vitro. Moreover, an increase in oocyte maturation rate per female was observed. Thus, the use of LANs could be useful to alleviate the effects of handling-stress during assisted reproductive procedures in wild ungulates.

Effect of egg yolk, cryoprotectant, and various sugars on semen cryopreservation in endangered Cuvier's gazelle (Gazella cuvieri).

Cryopreservation of spermatozoa from endangered species is a valuable tool for genetic management. Previous studies showed the feasibility of cryopreservation of spermatozoa from various endangered gazelles but have also revealed difficulties with available protocols for semen freezing in Cuvier's gazelle (Gazella cuvieri). Experiments were carried out to investigate the effect of (a) 5% or 20% egg yolk or 4% or 6% glycerol, and (b) addition of sugars (glucose, fructose, lactose and raffinose) on cryopreservation using a Tes-Tris-based diluent (TEST). A diluent containing 13.5% raffinose, 5% or 20% egg yolk, and 6% glycerol (REYG) was also evaluated. Semen was obtained by electroejaculation from 22 G. cuvieri males. Diluted samples were loaded into 0.25 ml straws, cooled to 5 degrees C over 1.5h (-0.16 degrees C/min), equilibrated at that temperature for 2h, frozen in nitrogen vapours for 10 min and plunged into liquid nitrogen. Subsamples were assessed for motility and acrosome integrity upon collection, after refrigeration-equilibration, after freezing and thawing, and 2h after thawing. Use of TEST with 20% egg yolk or with 4% glycerol led to worse motility preservation, whereas TEST with 5% egg yolk and 6% glycerol led to better results. Addition of fructose, lactose or raffinose to TEST resulted in similar or worse preservation of motility than inclusion of glucose. On the other hand, use of a raffinose-based medium with 20% egg yolk and 6% glycerol (REYG) afforded better preservation of motility than use of TEST. With REYG, 20% egg yolk was better than 5% egg yolk for motility preservation. Differences were noted between males in their responses to cryopreservation when using different egg yolk or glycerol concentrations. Moreover, spermatozoa from most males exhibited better cryopreservation with REYG although some were better cryopreserved in TEST. The raffinose-based diluent thus represents an improvement over previous results but more work is needed to better characterize cryopreservation conditions for future routine banking of Cuvier's gazelle spermatozoa.

In vitro oocyte maturation, fertilization and culture after ovum pick-up in an endangered gazelle (Gazella dama mhorr).

The recovery of immature oocytes followed by in vitro maturation, fertilization and culture (IVMFC) allows the rescue of biological material of great genetic value for the establishment of genetic resource banks of endangered species. Studies exist on sperm cryopreservation of endangered Mohor gazelle (Gazella dama mhorr), but no work has been carried out yet on oocyte collection, fertilization and culture in this or related species. The purpose of this study was to develop a protocol for ovarian stimulation for the recovery of oocytes and subsequent IVMFC in the Mohor gazelle using frozen-thawed spermatozoa. Ovum pick-up was performed after ovarian stimulation with a total dose of 5.28 mg of ovine FSH. A total of 35 oocytes were recovered from 56 punctured follicles (62%) (N=6 females). Out of 29 cumulus-oocyte complexes matured in vitro, 3% were found at germinal vesicle stage, 7% at metaphase I, 21% were degenerated, and 69% advanced to metaphase II. Fertilization and cleavage rates of matured oocytes were 40 and 30%, respectively. Embryos cleaved in vitro up to the 6-8 cell stage but none progressed to the blastocyst stage, suggesting the existence of a developmental block and the need to improve culture conditions. Although more studies are needed to improve hormonal stimulation and oocyte harvesting, as well as IVMFC conditions, this study demonstrates for the first time the feasibility of in vitro fertilization with frozen-thawed semen of in vitro matured oocytes collected by ovum pick-up from FSH-stimulated endangered gazelles.

Inbreeding and reproduction in endangered ungulates: preservation of genetic variation through the Organization of Genetic Resource Banks.

There is a constant increase in the number of species suffering marked reductions in population size. This reduction in size and the lack of genetic flow may lead to a decrease in genetic variability and to matings between close relatives (i.e. inbreeding) with an ensuing reduction in fitness. It is thus important to understand the mechanism underlying the deleterious effects of inbreeding and to develop reproductive biotechnologies that will allow the reduction of inbreeding depression by facilitating gene exchange between populations. The study of three endangered species of gazelles, Cuvier's gazelle (Gazella cuvieri), Mohor gazelle (Gazella dama mhorr) and dorcas gazelle (Gazella dorcas neglecta) has revealed that inbreeding negatively affects several semen parameters (motility, sperm morphology, acrosome integrity). Semen cryopreservation has been achieved in the three species but success varies depending on the diluent employed and the level of inbreeding. Artificial insemination of Mohor gazelles have led to the birth of the first gazelle born using frozen-thawed semen but improvements are needed before this technology can be applied on a routine basis for the genetic management of the populations. Collection of oocytes after ovarian stimulation, followed by in vitro maturation, fertilization and culture has met with some initial success in the Mohor gazelle. These, together with other reproductive technologies, will offer an invaluable help in preserving the maximum of genetic diversity of these and related endangered ungulate species.

Periparturient increase in faecal egg counts in a captive population of mohor gazelle (Gazella dama mhorr).

The objective of this study was to assess whether there was a periparturient rise in the faecal egg output of a population of North African gazelles (Gazella dama mhorr) kept in captivity in Almeria, southern Spain. In one experiment faeces were collected from 47 female gazelles on three days in winter, in November and December 1995 and January 1996; in a second experiment faecal samples were collected from nine pregnant gazelles at weekly intervals from July 1996 to June 1997. The mean trichostrongylid faecal egg counts were significantly higher (P < 0.05) in the periparturient gazelles than in the pregnant and non-pregnant animals only when the births took place in winter. Other factors, including the gazelle's age, its level of inbreeding, the number of previous births, and its trichostrongylid egg output at the beginning of the study did not affect whether it showed a periparturient rise. The parasites responsible for the rise were different in the two experiments.

Sperm cryopreservation in three species of endangered gazelles (Gazella cuvieri, G. dama mhorr, and G. dorcas neglecta).

Long-term storage of semen by cryopreservation, with high recovery rates on thawing, is essential for the establishment of genetic resource banks of endangered species. The purpose of the present study was to evaluate various diluents for the cryopreservation of spermatozoa from three species of gazelles (genus Gazella) in a captive breeding program. The diluents compared were Tes (N-tris(hydroxymethyl)methyl-2 aminoethane sulfonic acid)-Tris with 5% egg yolk and 6% glycerol (TEST) and Triladyl, yolk-citrate, Tris-trehalose, and Tris-lactose-all of them with 20% egg yolk and 6% (Triladyl) or 8% glycerol. Semen was obtained by electroejaculation from 12 G. cuvieri, 12 G. dama, and 13 G. dorcas males. Samples with less than 50% motile sperm, positive endosmosis, or acrosome integrity were not used. Diluted samples were loaded into 0.25-ml straws, cooled slowly to 5 degrees C over 1.5 h (-0.16 degrees C/min), equilibrated at that temperature for 2 h, frozen in nitrogen vapors for 10 min, and plunged into liquid nitrogen. Subsamples were assessed fresh, after refrigeration-equilibration, after freezing and thawing, and 2 h after thawing. Differences were seen between diluents, with best overall recovery rates after freezing and thawing found with Triladyl, TEST, and Tris-trehalose in G. cuvieri, TEST in G. dama, and Triladyl and TEST in G. dorcas. Differences were observed between species in the ability to withstand freezing and thawing, with best results seen in G. dorcas, intermediate results in G. dama, and worst results in G. cuvieri. These differences were inversely related to the average values of inbreeding of these populations. The underlying mechanism responsible for these differences may be a differential resistance to osmotic shock.

Effect of different methods of administration of ivermectin on its efficacy against the shedding of gastrointestinal nematode eggs by gazelles.

Seven different methods of administering ivermectin to gazelles were compared: subcutaneous injection, direct oral administration, administration in individual feeds, administration in a herd feed, direct oral administration of a second ivermectin formulation, administration in individual water supplies, and administration in the herd's water supply. The first five treatments were effective, as monitored by faecal egg count reduction tests, and administration in individual feeds or in a herd feed avoided the need to capture the animals, with the attendant risk of mortality. Of the factors associated with the recipients (species, sex, age and inbreeding coefficient) age was the only significant factor for the efficacy of the treatment. Oral or subcutaneous, individual or collective, and direct or indirect administrations were equally satisfactory for the treatment of all the parasite groups studied. Only when parasitic problems were due to Nematodirus species did direct administration to individual animals appear to be preferable.

Oral administration of mebendazole failed to reduce nematode egg shedding in captive African gazelles.

Idiosyncracies are observed in captive wild animals as regards the pharmacokinetics and efficacy of anthelmintics. This could be attributed to such factors as differences in host's metabolism, irregular distribution of anthelmintics due to the way they are administered and worm resistance to anthelmintics. Previously mebendazole was found to be poorly effective when administered in feed. An experiment was conducted to evaluate the efficacy of mebendazole when administered at the dosage rate of 15-20 mg/kg body weight to gastrointestinal nematodes in captive gazelles. Fifty-eight adult gazelles (Gazella cuvieri) were divided into four groups: T1 (animals dosed orally, directly into the mouth), T2 (treated orally, mixed in the water of a herd), T3 (treated orally, mixed in the water of one animal), and T4 (not treated). Individual faecal samples were taken before treatment, and 15 days thereafter. Mean percentages of reduction of egg shedding were calculated for Nematodirus spp., other trichostrongyles, total trichostrongylids, Trichuris spp. and total nematodes. No statistically significant differences were detected between the treatment groups and the control group or among the animals in the three treatment groups.

Abomasal and small intestinal nematodes from captive gazelles in Spain.

The abomasal and small intestinal helminth fauna of three species of captive gazelles (Gazella dama mhorr, G. cuvieri and G. dorcas neglecta) kept in captivity in Almería (southeast Spain) have been studied, and the following species were identified: Nematodirus spathiger, N. filicollis, N. helvetianus, Camelostrongylus mentulatus, Trichostrongylus vitrinus, T. probolurus, T. colubriformis, Ostertagia ostertagi, O. harrisi, Teladorsagia (Ostertagia) circumcincta, and T. (Ostertagia) davtiani. Camelostrongylus mentulatus and N. spathiger were the most prevalent and abundant parasites. Ostertagia ostertagi, O. harrisi, N. helvetianus, and T. (Ostertagia) davtiani were identified for the first time in the genus Gazella. In addition, O. harrisi and Trichostrongylus probolurus are new records for Spain.