Successful Captive Breeding Research Articles

Application of mitochondrial genes sequences for measuring the genetic diversity of Arabian oryx

Arabian oryx (Oryx leucoryx) had faced extinction in the wild more than three decades ago and was saved by the prudent efforts of captive breeding programs. A clear understanding of the molecular diversity of contemporary Arabian oryx population is important for the long term success of captive breeding and reintroduction of this potentially endangered species. We have sequenced the segments of mitochondrial DNA including12S rRNA, 16S rRNA, cytochrome b (Cyt-b) and control region (CR) genes of 24 captive-bred and reintroduced animals. Although the sequences of 12S rRNA, 16S rRNA and Cyt-b were found to be identical for all the samples, typical sequence variations in the CR gene were observed in the form of 7 haplotypes. One of these haplotypes has been reported earlier while the remaining 6 haplotypes are novel and represent different lineages from the founders. The haplotype and nucleotide diversities were found to be 0.789 and 0.009 respectively. The genetic distances among the 7 mtDNA haplotypes varied from 0.001 to 0.017. These findings are of potential relevance to the management of captive breeding programs for the conservation of Arabian oryx.

Reproductive Cycle Observation of the Okinawa Rail (Gallirallus okinawae) in the Wild

The captive breeding program of the Okinawa rail started in 2008. For successful captive breeding, information related to reproduction, such as age at sexual maturity, testicular cycles and ovulatory cycles, is essential to predict when reproduction is possible and when certain reproductive behaviors are most likely to occur. We made gross and histological observations of the reproductive organs of Okinawa rails to gain understanding of sexual maturity, the testicular cycle and the ovulatory cycle. We found that the weight of the testis was smallest in December and largest in March. Changes in the diameter of the seminiferous tubule showed the same pattern. Mature sperm were observed from March to June. The heaviest ovary was observed in April. A single peak of reproduction, from March to April, was observed in males and females. Our observations suggested that the Okinawa rail is a seasonal breeder. Establishing suitable breeding pairs will be critical to ensure success of the Okinawa rail captive breeding program. Our results suggested that pairing must be started before March. If supportive breeding is used, semen should be collected from March to June and artificial insemination conducted in April.

Comparative parasitological examination on sympatric equids in the Great Gobi “B” Strictly Protected Area, Mongolia

The Przewalski’s horse (Equus caballus przewalskii) became extinct in the wild during the 1960s. Based on a successful captive breeding program, Przewalski’s horses were reintroduced to the Great Gobi Part “B” strictly protected area (SPA) in SW Mongolia in the late 1990s. The Asiatic wild ass (Equus hemionus hemionus), Przewalski’s horse, and sometimes domestic horses live sympatricly in the Gobi B SPA. Previously published data demonstrates that, as a result of their different requirements and utilization of the park’s resources, their home-range size and social structure differ. Parasitological examinations in the three equid species show how the factors “home range, social structure, and resource selection” significantly impact parasitic burden. Asiatic wild asses are potentially exposed to a higher risk of parasite re-infection due to their temporal aggregation in very large groups. This study demonstrates a highly significant greater parasite load in the Asiatic wild ass for the majority of parasites evaluated (Dictyocaulus arnfieldi, Trichostrongylus axei, Strongyloides westeri, Parascaris equorum) compared to Przewalski’s horses and domestic horses in the same habitat. Domestic horses had higher parasite loads for eggs of strongylids, eggs of anoplocephalidae, and Eimeria leuckarti. The potential risk of cross infection between sympatric living equids is high, as is the cross infection between ruminants and equids. Furthermore, this study reports for the first time the occurrence of lungworms in free-ranging Przewalski’s horses. Whereas, Asiatic wild asses and Przewalski’s horses seem to cope very well with the sometimes high parasite burden, Mongolian domestic horses manifested typical parasite burden symptoms.

Cooperative conservation: zoos and in situ captive breeding for endangered Island fox Urocyon littoralis ssp

Successful captive breeding of endangered Island foxes, which essentially saved San Miguel Island foxes Urocyon littoralis littoralis and Santa Rosa Island foxes Urocyon littoralis santarosae from extinction, was facilitated by collaboration between land‐management agencies and the zoo community. Although the captive breeding was conducted in situ (on‐island) by the agencies, participating zoos offered skill sets and experience that the agencies lacked. Zoos contributed significantly to the development of Island fox husbandry guidelines, genetic management of captive foxes through a studbook, provision of adequate veterinary care and conservation education. Such proactive collaboration may become increasingly necessary for species conservation, given likely impacts to species from global climate change and human population growth.

Induced ovulation mimics the time-table of natural development in the stripe-faced dunnart, Sminthopsis macroura and results in the birth of fertile young

Induced ovulation maximizes captive breeding success, increasing productivity and facilitating the contribution of otherwise infertile animals to the genetic pool. In marsupials, induced ovulation to produce fertile young is unknown. Here we present an induction protocol efficient in inducing non-cycling and non-reproductive females to cycle, mate, ovulate, and conceive. Ovulation was induced in Sminthopsis macroura using an initial injection of 0.06 IU equine serum gonadotropin (eSG)/g (time 0), followed on day 4 by 0.04 IU eSG/g. Using this induction regime, the timing of follicular and embryonic development mimics natural cycles and results in the birth of viable, fertile young. Response to induction is not significantly affected by animal age, making this protocol an effective conservation tool. We have established a time-table of development following induction, providing a source of precisely timed research material. This is the first induced ovulation protocol in any marsupial to result in demonstrated fertile offspring and to allow the reliable collection of known-age samples during both the follicular phase and the gestation period.

Seasonal Profiles of Ovarian Activity in Iberian lynx (Lynx pardinus) Based on Urinary Hormone Metabolite Analyses

The Iberian Lynx Ex-Situ Conservation Programme is an essential part of a co-ordinated action plan to conserve the most endangered felid species of the world. Successful captive breeding demands reliable methods for reproduction monitoring including reliable non-invasive pregnancy diagnosis. During a 3-year study, urine samples from six captive Iberian lynx females were obtained (one non-pregnant, one pseudo-pregnant and 11 pregnant cycles). Progesterone, pregnanediol and oestradiol were determined in urinary extracts and relevant urinary oestrogen metabolites were characterized by high-performance liquid chromatography (HPLC). Urinary progestins did not follow the typical pregnancy-related course of felids. In the lynx, we failed to demonstrate an urinary progestin elevation during pregnancy. In contrast, urinary oestrogens increased from 3.8 +/- 0.6 to 8.6 +/- 0.5 ng/mg creatinine (p < 0.001) during the pregnancy. A comparison of pseudo-pregnant with pregnant cycles revealed a further increase of oestrogens caused by implantation (p < 0.05). In one female, which refused to mate, no difference was estimated between oestrogens levels during the breeding and non-breeding seasons. Almost 10-fold higher oestrogen concentrations were measured in urines of females that shared enclosures with males. HPLC analysis of oestrogens in urine samples collected from Iberian lynx during the pregnancy revealed that lynx urine is composed of two polar oestrogen metabolites in addition to oestrone and minor amounts of oestradiol. Oestrone was detectable in all urinary extracts (8-12% of metabolites), whereas oestradiol was elevated only during late pregnancy (18%). Thus, seasonal luteal activity in Iberian lynx can be monitored by urinary oestrogens. The increase of urinary oestradiol during late pregnancy might indicate an oestradiol secretion by the lynx placenta.

Successful captive breeding of Mitchell's water monitor, Varanus mitchelli (Mertens 1958), at Perth Zoo

Mitchell's water monitors (Varanus mitchelli) have been maintained on display at Perth Zoo since 1997. They are generally a timid species but have been maintained and bred in a mixed species water feature exhibit. In this article we describe their captive management and behavior with an insight into their reproductive biology. Between 2002 and 2005, 11 clutches were laid ranging from 13 to 27 (X = 20) eggs from one female. Egg size ranged between 3.00 and 6.08 g (X = 4.77 g) in weight, 22.8 and 31.9 mm (X = 28.3 mm) in length, and 11.1 and 19.3 mm (X = 17.1 mm) in width. Oviposition included double and triple clutches ranging between 41 and 60 days apart (X = 48 days), events n = 6. Four clutches were incubated at three different temperatures and hatchlings emerged after 157-289 days. The weight of the hatchlings ranged between 2.60 and 4.52 g (X = 4.34 g). Total length ranged between 140.1 and 178.0 mm (X = 165.9 mm) and snout-vent length ranged from 53.8 to 70.0 (X = 64.4 mm). Juvenile growth and development information is presented from hatching through to approximately 3 years of age.

A Chromosomal Inversion Unique to the Northern White-Cheeked Gibbon

The gibbon family belongs to the superfamily Hominoidea and includes 15 species divided into four genera. Each genus possesses a distinct karyotype with chromosome numbers varying from 38 to 52. This diversity is the result of numerous chromosomal changes that have accumulated during the evolution of the gibbon lineage, a quite unique feature in comparison with other hominoids and most of the other primates. Some gibbon species and subspecies rank among the most endangered primates in the world. Breeding programs can be extremely challenging and hybridization plays an important role within the factors responsible for the decline of captive gibbons. With less than 500 individuals left in the wild, the northern white-cheeked gibbon (Nomascus leucogenys leucogenys, NLE) is the most endangered primate in a successful captive breeding program. We present here the analysis of an inversion that we show being specific for the northern white-cheeked gibbon and can be used as one of the criteria to distinguish this subspecies from other gibbon taxa. The availability of the sequence spanning for one of the breakpoints of the inversion allows detecting it by a simple PCR test also on low quality DNA. Our results demonstrate the important role of genomics in providing tools for conservation efforts.

Non‐invasive Monitoring of Hormones: A Tool to Improve Reproduction in Captive Breeding of the Eurasian Lynx

The survival of many critical endangered mammal species is often depending on successful captive breeding programmes which include the future option of reintroduction to the wild. Breeding in captivity also demands the application of modern assisted reproductive techniques to ensure maximal biodiversity, but knowledge on reproductive physiology is often limited. Therefore, non-invasive monitoring of urinary and faecal hormones has become an important tool for reproductive management. To exemplify the importance of non-invasive hormone monitoring, we choose the Eurasian lynx as a model for the world's most endangered felid species, the Iberian lynx. We analysed faecal samples of pregnant and pseudo-pregnant female Eurasian lynxes during a 3-year study period. Compared to pre-mating levels faecal progesterone metabolite profiles revealed a tendency towards higher levels in pregnant and pseudo-pregnant females with no difference between both categories. Oestrogen levels raised in both pregnant and pseudo-pregnant females with a tendency to be more elevated and prolonged in pregnant females. Surprisingly both E2 and P4 metabolites were highly correlated (r(2) =0.8131, p < 0.0001) showing a postpartum increase both in pregnant and pseudo-pregnant females. The results from the Eurasian lynx revealed that the measurement of faecal progesterone metabolites led to profiles dissimilar to profiles shown in other felid species, but similar to those from faecal gestagen metabolite analysis in the Iberian lynx. To identify faecal gestagen and oestrogen metabolites a radio-metabolism study was performed. Using the progesterone immunoassay two major progesterone metabolites were detected demonstrating that the assay indeed tracks the relevant metabolites. The oestrogen assay measured authentic 17beta-oestradiol and oestrone, and their conjugates. The analysis of the faecal metabolite composition in samples from early and late pregnancy and lactation particularly revealed a distinct shift in the relation between 17beta-oestradiol and oestrone that changed in favour of oestrone. This might indicate different hormone sources during and after pregnancy (corpus luteum, placenta). We hypothesize, that placental steroid analysis in combination with other highly sophisticated analytical techniques, like liquid chromatography mass spectrometry or urinary relaxin analysis may led to analytical options to confirm pregnancy and to differentiate this from pseudo-pregnancy in lynx species.

Assessment of Fertility Status in the Male Sumatran Rhino at the Sumateran Rhino Sanctuary, Way Kambas National Park, Lampung

Sumatran rhino is the most endangered rhino species. Its population is estimated less than 300 individuals remaining in the wild with highly declining rate to 50% in the last 15 years. The number of male rhinoceroses in the captivity are very few, therefore the assessment of its fertility is very important in order to support the breeding success since the captive breeding success is very poor. The objectives of this study were (i) to determine the male reproductive status, (ii) to establish a reliable semen collection method, and (iii) to assess semen parameters of the fresh collected sample. Three methods of semen collection were examined to determine its fertilizing potential, i.e. (i) stimulated combination of artificial vagina (AV), penile massage (PM) and accessory gland massage (AGM); (ii) AV and PM; and (iii) only with PM. The first method gave the best result with an ejaculation success of 85.71% (6/7, n = 7). The second and third methods obtained an ejaculation success rate of 50% (2/4, n = 4)) and 25% (1/4, n = 4), respectively. The collected ejaculates had a volume of 1.2-12.4 ml with whitish to cream turbid colour and pH 6.90-6.99. Sperm concentration was (143-333) x 103 sperm/ml. The quality of the sperm was low with only approximately 1% of them moved forward slowly. Approximately 80% of the spermatozoa were immature (prox. cytoplasmic droplet) with head (macro-, microcephalic) and tail abnormalities (broken tail). Semen quality increased after several collections and the amount of immature sperm decreased up to 5%. Electroejaculation procedure could produce 34 ml semen, but no sperm was found in the ejaculate. Hence, the combination of AV, PM, and AGM could get higher volume of ejaculate compared to other methods, but sperm concentration was better obtained using AV and PM only. Repeated semen collection increased semen quality, although the male has low fertilizing capacity due to low sperm concentration (oligozoospermia) and small volume of the ejaculate (oligospermia). Key words: Sumatran rhino, semen, sperms, collection

Aufzucht von Chilemeerespelikane ( Pelecanus occidentalis thagus) im Vogelpark Walsrode mit Vergleichen zu anderen publizierten Erfahrungen mit Pelikanen

The probable world first captive breeding success in Chilean Brown Pelican ( Pelecanus occidentalis thagus) in captivity is reported. Two chicks were hand reared at Walsrode Birdpark, Germany in 2006. Parents were imported in October 2005 and were wild birds that had been injured and subsequently rescued. All 10 adult birds demonstrated detrimental and chronically handicaps. First eggs were laid in January 2006. Eggs were incubated artificially at 37.1 °C and 56% rel. humididty. Two chicks hatched after 33 days of incubation. First feeding was made one day after hatching in the morning. Food was given 5 times a day. Until the 8th day after hatching, exclusively cut naked baby rats were fed. For stimulating the digestion and providing a good intestinal flora Lactobacillus powder was used and food was warmed up before application. After 8 days the diet was changed to fish. For development see figures. Worth to mention is the black-purple colour of the freshly hatched chicks which became successively paler with age. Also the development of the feather areas is characteristic for this (sub)-species. Taxonomy is discussed with some morphological arguments that are typical for subspecies. Nevertheless, some indications of possible breeding barriers between subspecies could support the suggestions for species status. Hatching and rearing data are compared with dates from other zoos and species of pelicans. The good development of the Walsrode birds based on the data and methods described leaves this article useful as a reference guideline for hand rearing pelicans.

Increasing the chances of successful reintroduction of white-clawed crayfish (Austropotamobius pallipes) in the Peak District National Park, UK

The current strategy for conserving white-clawed crayfish, Austropotamobius pallipes, contained in the United Kingdom Biodiversity Action Plan includes reintroduction to isolated water, free of signal crayfish, Pacifastacus leniusculus. The purpose of this project was to improve the chances of successful reintroduction of white-clawed crayfish into the River Lathkill in the Peak District National Park and to demonstrate how the method might be applicable to other reintroduction and captive breeding schemes. The method focussed on the habitat requirements of white-clawed crayfish, in particular the need of younger crayfish to conceal themselves from predators. Cages were built which provided juvenile crayfish with hides. The cages were tested in the river and juvenile growth and survival were recorded. Adult crayfish survival was only 50% annually whilst survival of juveniles was much less. Despite this, the habitat within the cages protected crayfish during moult and is thought to have increased their survival rate. The project has provided evidence for development of a successful reintroduction and captive breeding programme for white-clawed crayfish that will enable the conservation strategy to be undertaken more effectively.

Characterisation of the sympathetic nervous system of Asian ( Elephas maximus) and African ( Loxodonta africana) elephants based on urinary catecholamine analyses

Assessing the welfare status of captive animals using non-invasive measurements of hormones is of growing interest because this can serve as an effective tool to facilitate the optimization of environmental and husbandry conditions. Both the African elephant ( Loxodonta africana) and the Asian elephant ( Elephas maximus) exhibit extremely low breeding success in captivity, and because elevated levels of stress may negatively influence reproductive functions, this study sought to establish a method for assessing sympathoadrenal activity in captive female elephants. We found a circadian variation in urinary noradrenaline (norepinephrine, NE), adrenaline (epinephrine, Epi) and dopamine (DA) under short day length. Peak activity of noradrenaline and dopamine was noted at 3 a.m. Adrenaline showed a biphasic pattern with a minor peak recorded at 3 a.m. and a major peak 9 a.m. Under long-day photoperiodic conditions, simultaneous peaks of noradrenaline and adrenaline were again noted at 3 a.m. whereas dopamine does not appear to have a distinct circadian pattern under long-day length. A transfer of two elephant cows resulted in a marked increase in urinary adrenaline and noradrenaline levels, confirming that the transfer represented a stressful event. During the peripartal period, noradrenaline concentrations increased and maximum concentrations were obtained at delivery. Daily measurements of urinary dopamine throughout the follicular phase revealed an increase in dopamine secretion close to ovulation. This increase might indicate a role of dopamine in the ovulatory mechanisms. These results suggest that changes in urinary catecholamine excretion reflect fluctuations in sympathoadrenal activity and may be a useful indicator of stress.

Oestrous cycle of captive southern hairy-nosed wombats ( Lasiorhinus latifrons) in South Australia, Australia

There is limited information available on the oestrous cycle of female southern hairy-nosed wombats ( Lasiorhinus latifrons). This is mainly due to an extremely poor breeding success in captivity and the difficulty in routine recapturing of these cryptic, semi-fossorial animals in the wild. The aim of this study was to characterise the oestrous cycle of this species by monitoring peripheral plasma concentrations of progesterone and oestradiol, assessing changes in vaginal cytology, pouch condition and the urogenital sinus. Eight adult female wombats were monitored during the breeding season (July–December) over 2 years (2002–2003). Samples were collected up to three times a week. Vaginal smears contained several cell types, categorised by morphology, as either superficial epithelial cells or parabasal–intermediate cells. Leucocytes were also counted. Plasma progesterone profiles showed a mean oestrous cycle length of 36.33 ± 0.67 days with a peak progesterone concentration of 139.53 ± 10.62 nmol/L. Levels of oestradiol peaked at a mean level of 467.33 ± 44.32 pmol/L on average 5 days before a rise in plasma progesterone values. The proportion of epithelial cells in vaginal smears varied throughout the cycle, with a high percentage of superficial epithelial cells observed during the follicular phase. During periods when progesterone concentrations were high, a greater percentage of parabasal–intermediate cells was observed. In conclusion, this study has characterised the oestrous cycle of the southern hairy-nosed wombat and confirmed that changes in vaginal smears together with pouch and urogenital sinus details could be used to determine signs of oestrus in this species.

Husbandry of wild-caught greater sage-grouse

This study reports the first successful husbandry and breeding in captivity of wild-caught greater sage-grouse (Centrocercus urophasianus). In October 2003, 21 hatch-year greater sage-grouse were trapped in northwestern Nevada and transported to Fort Collins, Colorado. We held grouse in pens at the United States Department of Agriculture's National Wildlife Research Center for 8 months. We offered a varied diet, including native food items such as sagebrush (Artemisia tridentata and A. tripartita) and yarrow (Achillea millefolium). We housed grouse in a large flight pen and allowed to them free-range as one flock. Mortality rate was 16.7%. Several of the grouse exhibited breeding behavior, and 13 eggs were laid. We describe the techniques used to house and feed wild-caught sage-grouse. This study has conservation implications for captive breeding of this species of concern.

Reproductive parameters and life-history variables in captive golden-bellied mangabeys (Cercocebus agilis chrysogaster).

An understanding of the reproductive physiology of captive primates is vital for their successful management. We report on reproductive parameters and life-history variables collected at the Sacramento Zoo for five female golden-bellied mangabeys (Cercocebus agilis chrysogaster) over a 7-year period. For each female, we collected data on their sexual skins, menses, gestation, and other pregnancy-related factors, such as postconception bleeding and swelling. We used these data to estimate life-history variables, such as age at onset of estrus, menses, and conception, as well as the duration of intervals between births, and between parturition and the resumption of cycling. Cercocebus agilis chrysogaster is comparable to other Cercocebus species in terms of reproductive parameters, although variability is exhibited within and among female subjects. In some cases, it appears that stressful incidents altered cycle length or halted cycling altogether. We suggest the use of husbandry practices that include consistent data collection to enhance regularity in reproductive cycles, and maximize captive breeding success for this rare species.

Assessment and conservation of the giant gomeran lizard (Gaiotia bravoana)

In June 1999, a species of lizard previously considered extinct was rediscovered on the island of La Gomera, Spain. The giant gomeran lizard, Galotia bravoana, is highly endangered and at...

Effects of Environmental Stress on the Parental Behavior in Crested Ibis (Nipponia nippon)

When maintained in captivity in 'intensive aviaries' that offered suboptimal and unnatural living conditions, parent Crested Ibis successfully incubated their eggs but subsequently killed the chicks and discarded them at the point of hatching. In order to investigate this aberrant behavior, new 'semi-natural' breeding cages were established in natural woodland habitat, and the parental behaviors of five breeding pairs from F1 and F2 generations were examined using video recordings. Over the three-year study period, 13 chicks hatched and were successfully raised by the parents in the semi-natural cages, whereas no parental care was provided to those chicks that hatched in the intensive aviaries, and all perished. This experimental evidence revealed that their killing of the chicks might be a consequence of environmental stress caused by their inability to adapt to the artificial environment of an unnatural enclosure. We concluded that the environmental conditions under which they are maintained is a crucial factor for the behavioral adaptation and captive breeding success of this endangered species.

Towards the effective management of the Arabian oryxOryx leucoryx in the Kingdom of Saudi Arabia

Traditionally the distribution of the Arabian oryx in the Kingdom of Saudi Arabia has been closely associated with that of the major sand deserts, notably the Great Nafud in the north and the Rub' al Khali in the south. Due to a successful captive breeding programme the Arabian oryx was reintroduced in the ‘Uruq Bani Ma’arid Protected Area, on the western edge of the Rub' al Khali during 1995, thereby re-establishing the southern population in the Kingdom of Saudi Arabia. Population viability analysis is used to demonstrate the re-establishment of the northern oryx population, the vulnerability of certain populations and to suggest a management approach that should enhance the survival of the free-living populations in the Kingdom of Saudi Arabia. Population models predict that the ‘Uruq Bani Ma’ arid population could be subject to extinction (Psurvival=0.38) when modeled in isolation. Associated with this is a low heterozygosity. The proposed release strategy of oryxes into a northern protected area results in maximum population growth during the first 20-years and continued positive population growth during the 100-year modeling period. The latter free-living population is, however, also subject to extinction due to its relatively small size. By managing all of the Arabian oryx populations in the Kingdom of Saudi Arabia as a single metapopulation, survival of the free-living populations are enhanced, without negatively affecting the other populations.

Demography of the California Condor: Implications for Reestablishment

Abstract: The remnant wild population of California Condors (Gymnogyps californianus) of the 1980s exhibited a rapid population decline caused by high mortality rates among adult and immature birds. The most prominent mortality factor was lead poisoning resulting from ingestion of bullet fragments in carcasses. Successful captive breeding has allowed many birds to be released to the wild since 1992, based originally on an assumption that exposure to lead could be prevented by food subsidy. The mortality of released birds, however, has generally exceeded levels needed for population stability calculated from simple population models. Collision with overhead wires was the most frequent cause of death in releases before 1994. Lead poisoning again surfaced as a problem starting in 1997 as older birds began feeding on carcasses outside the subsidy program. Although poisonings have been treated successfully by chelation therapy in recaptured birds, food subsidy is proving an ineffective solution to lead exposure. The best long‐term solution appears to be either the creation of large reserves where hunting is prohibited or the restriction of hunting to nontoxic ammunition in release areas. Until sources of lead contamination are effectively countered, releases cannot be expected to result in viable populations. In addition, problems involving human‐oriented behavior have resulted in the permanent removal of many released birds from the wild. The most promising reduction in human‐oriented behavior has been achieved in one release of aversively conditioned, parent‐reared birds. Rigorous evaluation of the factors reducing attraction to humans and human structures has been hampered by confounding of techniques in releases. Behavioral problems could be more quickly overcome by adoption of a comprehensive experimental approach.