Catherine A. Herbert

Fertility control in female eastern grey kangaroos using the GnRH agonist deslorelin. 2. Effects on behaviour

In recent years fertility control has been proposed as an ethically acceptable alternative to lethal control techniques when managing overabundant kangaroo populations. A promising non-steroidal, non-immunological approach to contraception in female kangaroos involves the use of slow-release implants containing the gonadotrophin-releasing hormone (GnRH) agonist deslorelin. The practicality of using deslorelin implants as a management option is dependant on its effective inhibition of reproduction without negative physical or behavioural side-effects. This study investigated the behavioural effects of deslorelin implants in female eastern grey kangaroos. Treatment had no detectable effects on crepuscular activity. Alterations in the frequency of sexual interactions were observed in deslorelin-treated females, with a behavioural oestrus induced ~3 days after combined removal of pouch young and deslorelin administration. Copulation was observed during this early oestrous period, but conception was not achieved and pouch young were not observed in any treated females. Control females gave birth within 69.6 ± 10.4 days (mean ± s.e.m., n = 9) of placebo implant administration. The first births observed in treated animals were on Days 510, 637 and 643 after treatment. The remaining seven treated animals had not bred by the end of the study, a period of 647 days.

Effects of a Gonadotropin-Releasing Hormone Agonist Implant on Reproduction in a Male Marsupial, Macropus eugenii

Abstract This study evaluated the potential of slow-release GnRH agonist (deslorelin) implants to inhibit reproductive function in the male tammar wallaby. The specific aim was to measure the effects of graded dosages of deslorelin on testes size and plasma LH and testosterone concentrations. Adult male tammar wallabies were assigned to four groups (n = 6 per group) and received the following treatment: control, placebo implant; low dose, 5 mg deslorelin; medium dose, 10 mg; high dose, 20 mg. All dosages of deslorelin induced acute increases (P < 0.001) in plasma LH and testosterone concentrations within 2 h, with concentrations remaining elevated during the first 24 h but returning to pretreatment levels by Day 7. Thereafter, there was no evidence of a treatment-induced decline in plasma testosterone concentrations. There was no detectable difference in basal LH concentrations between treated and control animals, nor was there a significant change in testes width or length (P > 0.05). These results suggest that the male tammar wallaby is resistant to the contraceptive effects of chronic GnRH agonist treatment. Despite the maintenance of testosterone secretion, the majority of male tammars (10 of 17) failed to respond to a GnRH challenge with a release of LH between Days 186 and 197 of treatment. The failure of animals to respond to exogenous GnRH suggests a direct effect of deslorelin on the pituitary, resulting in a level of desensitization that was sufficient to inhibit a LH surge but insufficient to inhibit basal LH secretion. The variation between animals is believed to result from earlier recovery of some individuals, in particular those that received a lower dose, or individual resistance to the desensitization process.

Inbreeding and testicular abnormalities in a bottlenecked population of koalas (Phascolarctos cinereus)

Habitat destruction and fragmentation, interactions with introduced species or the relocation of animals to form new populations for conservation purposes may result in a multiplication of population bottlenecks. Examples are the translocations of koalas to French Island and its derivative Kangaroo Island population, with both populations established as insurance policies against koala extinction. In terms of population size, these conservation programs were success stories. However, the genetic story could be different. We conducted a genetic investigation of French and Kangaroo Island koalas by using 15 microsatellite markers, 11 of which are described here for the first time. The results confirm very low genetic diversity. French Island koalas have 3.8 alleles per locus and Kangaroo Island koalas 2.4. The present study found a 19% incidence of testicular abnormality in kangaroo Island animals. Internal relatedness, an individual inbreeding coefficient, was not significantly different in koalas with testicular abnormalities from that in other males, suggesting the condition is not related to recent inbreeding. It could instead result from an unfortunate selection of founder individuals carrying alleles for testicular abnormalities, followed by a subsequent increase in these alleles’ frequencies through genetic drift and small population-related inefficiency of selection. Given the low diversity and possible high prevalence of deleterious alleles, the genetic viability of the population remains uncertain, despite its exponential growth so far. This stands as a warning to other introductions for conservation reasons.

SEROLOGIC SURVEY FOR TOXOPLASMA GONDII AND NEOSPORA CANINUM IN THE COMMON BRUSHTAIL POSSUM (TRICHOSURUS VULPECULA) FROM URBAN SYDNEY, AUSTRALIA

The common brushtail possum (Trichosurus vulpecula) has well adapted to increasing urbanization, resulting in greater interaction with humans and their domestic pets. Wildlife species in urban areas face a higher risk of exposure to zoonotic pathogens and may be affected by parasites hosted by cats (Toxoplasma gondii) or dogs (Neospora caninum), yet it is unknown to what extent urban T. vulpecula are exposed to these parasites. Antibodies to T. gondii and N. caninum were assayed in sera of 142 adult possums from the city of Sydney, Australia. Using the modified agglutination test, antibodies to T. gondii were found in 9 (6.3%) of the 142 animals in titers of 1:25 (4), 1:50 (1), 1:100 (1), 1:800 (1), 1:3,200 (1), 1:6,400 (1), and 1:12,800 (1). Of some T. vulpecula multiple sera samples within a 2-yr frame could be collected, but seropositive animals in general were not recaptured after initial seroconversion. One possum had a high T. gondii titer on 2 consecutive bleedings, 14 mo apart, and seropositive possums appeared normal when captured. Sex seemed not to have an affect on antibody prevalence, but age and location may play a role. Antibodies to N. caninum were not detected in 1:25 dilution of sera in the N. caninum agglutination test, indicating that T. vulpecula may not have been exposed to this parasite. This is the first serological survey for T. gondii and N. caninum infections in urban T. vulpecula.

Effects of deslorelin implants on reproduction in the common brushtail possum (Trichosurus vulpecula)

The present study investigated the effects of slow-release implants containing the gonadotrophin-releasing hormone (GnRH) agonist deslorelin on reproduction in the common brushtail possum (Trichosurus vulpecula). Captive female brushtail possums were assigned to control (placebo implant), low dose (4.7 mg deslorelin) or high dose (9.4 mg deslorelin) groups; males were assigned to control or high dose (9.4 mg deslorelin) groups. The acute effects of deslorelin treatment at the level of the pituitary gland were similar between the two sexes, where a transient rise in luteinising hormone concentration was induced over the first 24 h. In females, this was associated with the disruption of the normal oestrous cycle and mating within 2-10 days in some treated individuals, but no young were subsequently detected. By 3 weeks after treatment, treated females became anoestrus and remained infertile for at least one breeding season. The effects of treatment were reversible in a subset of females that had their implants removed, although the time taken to produce offspring was variable. Paradoxically, male brushtail possums remained fertile during chronic deslorelin exposure. Despite significant declines in basal follicle-stimulating hormone and testosterone concentrations, as well as an inability to respond to a GnRH challenge, treated males sired as many offspring as control males and there was no evidence of testicular regression. In conclusion, there is potential to control reproduction in female brushtail possums by using chronic GnRH agonist treatment.

Dominance, body size and internal relatedness influence male reproductive success in eastern grey kangaroos (Macropus giganteus)

Knowledge of the determinants of reproductive success is essential for understanding the adaptive significance of particular traits. The present study examined whether particular behavioural, morphological, physiological or genetic traits were correlated with male dominance and reproductive success using three semi-free-ranging captive populations (n = 98) of the eastern grey kangaroo (Macropus giganteus). The morphological traits measured included bodyweight, head, forearm, tail, pes and leg length, forearm and bicep circumference, and testis size. Blood samples were collected to determine serum testosterone concentrations. All individuals were typed for 10 microsatellite loci and paternity determined for each pouch young. To determine the influence of relatedness and genetic diversity on male reproductive success, internal relatedness, standardised heterozygosity and mean d(2) were calculated. Dominant males sired a significantly higher proportion of offspring than smaller, lower-ranked males and had higher testosterone concentrations. Males that sired offspring were significantly heavier and had larger body size. Sires were significantly more heterozygous and genetically dissimilar to breeding females than non-sires. Despite the wealth of knowledge on the social organisation of kangaroos, this is the first study to assign parentage and male reproductive success using molecular evidence.

Genetic consequences of isolation: island tammar wallaby (Macropus eugenii) populations and the conservation of threatened species

Isolation and restricted gene flow can lead to genetic deterioration in populations. Populations of many species are increasingly becoming fragmented due to human impacts and active management is required to prevent further extinctions. Islands provide an ideal location to protect species from many mainland threatening processes such as habitat loss and fragmentation, disease and competition/predation from introduced species. However their isolation and small population size renders them prone to loss of genetic diversity and to inbreeding. This study examined two endemic and one introduced population of tammar wallaby (Macropus eugenii) on three islands in the Houtman Abrolhos Archipelago, Western Australia: East Wallabi (EWI), West Wallabi (WWI) and North Islands (NI). Nine autosomal and four Y-linked microsatellite loci, and sequence data from the mitochondrial DNA (mtDNA) control region were used to examine the impact of long-term isolation (EWI and WWI) and small founder size (NI) on genetic diversity and inbreeding. This study found all three populations had low genetic diversity, high levels of effective inbreeding and increased frequency of morphological abnormalities. Isolation has also led to significant inter-population genetic differentiation. These results highlight the importance of incorporating genetic management strategies when utilising islands as refuges for declining mainland populations.

Leptospirosis serology in the Common Brushtail Possum (Trichosurus vulpecula) from urban Sydney, Australia

The common brushtail possum (Trichosurus vulpecula) is indeed a common marsupial in major cities of Australia. This species is known to be susceptible to leptospirosis and often lives in close contact with humans, raising concerns about the potential for transmission of this disease in urban areas. A total of 192 brushtail possum blood samples were collected from 136 individuals in suburban areas of metropolitan Sydney from November 2002 to November 2004. Sera were screened against a reference panel of 21 Leptospira spp. using the microscopic agglutination test. Leptospiral antibodies were detected in 9.6% (13/136) of tested brushtail possums and represented two serovars; antibodies to Leptospira interrogans serovar Hardjo were most frequently identified (11/136). A representative of the exotic serogroup Ballum, most likely serovar Arborea, was found in two of 136 brushtail possums. Exposure to leptospirosis seemed to be associated with age, as older animals had a higher incidence, but there was no distinction in relation to gender. Antibody prevalence varied between the different sampling sites and seropositive animals were clustered and restricted to a few sites. These data support the possible role of brushtail possums as a maintenance host for Leptospira spp. in urban environments and also identified them as a previously unknown and potential source of serovar Arborea.

High possum mortality on urban roads: implications for the population viability of the common brushtail and the common ringtail possum

Animal–vehicle collisions impact wildlife populations and in the northern suburbs of Sydney, both the common brushtail possum (Trichosurus vulpecula) and the common ringtail possum (Pseudocheirus peregrinus) are killed in large numbers. Over a two-year period almost 600 road-killed possums were observed from 217 road surveys covering over 7800 km, equating to 5.45 possums per week over the 36-km study area. Surveys were conducted along roads where the environment ranged from low-rise suburban to continuous sclerophyll forest. Significantly more ringtail possums were observed as road-kill, outnumbering brushtail possums by almost three to one. No sex bias was found amongst road-killed ringtail possums; however, seasonal trends and weight fluctuations were significant, with both males and females being at their lowest weights during the autumn breeding season. For brushtail possums a bias towards young males was observed. Sex and weight of road-killed brushtail possums were comparable to live-trap data from a previous study in the same location. In areas of such high possum mortality, wildlife managers may need to take action to mitigate possum road-kill.

Brushtail possums (Trichosurus vulpecula) in metropolitan Sydney: population biology and response to Suprelorin contraceptive implants

Abstract. The cohabitation of the common brushtail possum (Trichosurus vulpecula) with people in urban areas often causes conflict. Basic biological parameters are needed to evaluate potential new management options such as fertility control. This study investigated the biology of an urban brushtail possum population and the effects of Suprelorin contraceptive implants on individual females within that population. Trapping success remained constant over time, with 2.8 ± 0.2 individuals trapped per residential property. Recapture rates for males declined rapidly over time, with only 30% of males recaptured on the same property 12 months after initial capture (n = 30) and no males recaptured after 18 months (compared with female recapture rates of 58% after 18 months, n = 33). These data, combined with the preponderance of males observed within the lower age classes, suggests that male possum turnover is high within urban areas. This may be partially compensated for by the male-bias (1.8 : 1) observed in pouch young. Breeding was seasonal with the main peak of births in autumn, and a secondary smaller peak in spring. Suprelorin contraceptive treatment effectively inhibited reproduction in adult females for a minimum duration of 519 ± 7 (n = 5) and ≥700 ± 20 (n = 5) days after administration of one or two 4.7-mg implants, respectively, with no negative side-effects obvious. The concurrent collection of data on contraceptive efficacy and population-specific life-history parameters provides a unique opportunity to highlight the importance of understanding local population dynamics when evaluating the likely efficacy and implementation of fertility control programs to manage problem wildlife.