Accounting for cryptic population substructure enhances detection of inbreeding depression with genomic inbreeding coefficients: an example from a critically endangered marsupial

Characterizing inbreeding depression in wildlife populations can be critical to their conservation. Coefficients of individual inbreeding can be estimated from genome‐wide marker data. The degree to which sensitivity of inbreeding coefficients to population genetic substructure alters estimates of inbreeding depression in wild populations is not well understood. Using generalized linear models, we tested the power of two frequently used inbreeding coefficients that are calculated from genome‐wide SNP markers, F_H and F^^III, to predict four fitness traits estimated over two decades in an isolated population of the critically endangered Leadbeater's possum. F_H estimates inbreeding as excess observed homozygotes relative to equilibrium expectations, whereas F^^III quantifies allelic similarity between the gametes that formed an individual, and upweights rare homozygotes. We estimated F_H and F^^III from 1,575 genome‐wide SNP loci in individuals with fitness trait data (N = 179–237 per trait), and computed revised coefficients, F_H^{by group} and F^^{IIIby group}, adjusted for population genetic substructure by calculating them separately within two different genetic groups of individuals identified in the population. Using F_H or F^^III in the models, inbreeding depression was detected for survival to sexual maturity, longevity and whether individuals bred during their lifetime. F^^{IIIby group} (but not F_H^{by group}) additionally revealed significant inbreeding depression for lifetime reproductive output (total offspring assigned to each individual). Estimates of numbers of lethal equivalents indicated substantial inbreeding load, but differing between inbreeding estimators. Inbreeding depression, declining population size, and low and declining genetic diversity suggest that genetic rescue may assist in preventing extinction of this unique Leadbeater's possum population.     

Isolated remnant or recent introduction? Estimating the provenance of Yellingbo Leadbeater's possums by genetic analysis and bottleneck simulation

Effective conservation management requires that genetically divergent populations potentially harbouring important local adaptations be identified and maintained as separate management units. In the case of the endangered Australian Leadbeater's possum (Gymnobelideus leadbeateri), an arboreal marsupial endemic to Victoria, uncertainty over the evolutionary origin of a potentially important extant wild population recently discovered in atypical habitat (lowland swamp) at Yellingbo is hampering such efforts. The population is rumoured to be a recent introduction. Microsatellite allele frequencies at Yellingbo differed substantially from those in sampled populations in montane ash forest (F_ST between 0.23 and 0.36), and Bayesian clustering analyses of genotypes strongly separated them (K = 2). We conducted a suite of bottlenecking tests which all indicated that Yellingbo had undergone a recent reduction in size. The extent to which the distinctiveness of Yellingbo animals might be expected solely through bottlenecking associated with a recent introduction, was tested by simulating population–history scenarios seeded with genotypes from candidate wild and captive sources. No bottleneck scenario reproduced anything approaching the genetic distinction of the Yellingbo population, with all structure analyses placing Yellingbo in a separate cluster to simulated populations (K = 2, minimum F_ST = 0.13). These results suggest that Yellingbo does not share recent ancestry with other extant populations and instead may be a remnant of an otherwise extinct gene pool. Importantly, this may include genes involved in adaptation to a lowland swamp environment, substantially adding to the conservation importance of this population, and suggesting that separate management may be prudent until evidence suggests otherwise.     

Patterns of co-occurrence among arboreal marsupials in the forests of central Victoria,southeastern Australia

Abstract Patterns of co-occurrence were examined among the suite of species of arboreal marsupials that inhabit the montane ash forests of the Central Highlands of Victoria, southeastern Australia. The data used in our analyses were counts of different species of arboreal marsupials derived from stagwatching surveys of 152 sites throughout the Central Highlands region. Our work investigated both the number of species that tend to occur together, as well as identity of the taxa in such associations. A total of eight different species of arboreal marsupials were recorded from field surveys of montane ash forests: Leadbeater's Possum (Gymnobelideus leadbeateri), Sugar Glider (Petaurus breviceps), Yellow-bellied Glider (Petaurus australis), Greater Glider (Petauroides volans), Mountain Brushtail Possum (Trichosurus caninus), Feathertail Glider (Acrobates pygmaeus), Common Ringtail Possum (Pseudocheirusperegrinus), and Eastern Pygmy Possum (Cercartetus nanus). Large assemblages of taxa were rare and no sites supported the full array of species of arboreal marsupials– the maximum number of species recorded on a given 3 ha site was five. The results of our analyses indicated that if any one species of arboreal marsupials was present at a site, then on average 1. 5 other taxa will co-inhabit that site. The exception was the Common Ringtail Possum. When this species was detected, few other arboreal marsupial taxa tended to be recorded with it. Principal Co-ordinates Analysis showed that although combinations of animals comprised of either Mountain Brushtail Possum, Leadbeater's Possum, or Greater Glider were often recorded, there were no strong patterns in the identities of co-occurring taxa. This finding, together with the result that most taxa in these forests typically co-occurred with one or two other species, indicates that none of the arboreal marsupials inhabiting the montane ash forests of the Central Highlands of Victoria would be a good surrogate for (or ‘indicator’ of), the presence of the suite of other species in the assemblage.     

How many possums make a cow?

The importance of possums as competitors with livestock for pasture is sometimes used as justification for possum control. Unfortunately a confusion of wet and dry weight values of pasture and daily consumption by possums appears to have resulted in a significant overestimate of the economic costs of possums as pasture pests. A correctly estimated daily dry matter intake of pasture by possums is 0.0144 kg pasture dry matter per possum per day. For a possum density of 1 ha^-1, this amounts to a reduction of about 1% of a stock unit ha^-1 year^-1. At higher possum densities, which often occur on farms adjacent to forest or scrub, losses would be correspondingly higher. A more direct way to measure possum impact on pasture production would be to use exclosures.     

The formulation of a national strategy for biological control of possums and bovine Tb

Abstract

The Government has designated the control of possums and bovine Tb a National Science Strategy (NSS) research area. An NSS Committee has been appointed by the Ministry of Research Science and Technology, commissioned to “identify, coordinate and promote national priorities for possum and tuberculosis (Mycobacterium bovis) related research in order that threats both to New Zealand's export markets and to conservation values can be eliminated”.     

Mistletoe (

The condition of 79 plants of the loranthaceous mistletoe Tupeia antarcticain a podocarp-hardwood forest in the central North Island, New Zealand, was monitored over 4 years during a period of increasing possum density, following previous possum control. Mistletoe comprised 1.2% of total possum diet during the three years following possum control. Incidence of possum browse on mistletoe plants increased from 2.6% of plants when the trap-catch index of possum density was < 3%, to 75.9% of plants when trap-catch rates reached 4.6%. Mistletoe foliage cover declined from 49.8% to 15.6% and mean plant size declined by about 55% over the same period. The mistletoe population was dominated by plants with large haustoria, located in heavily shaded locations in the lower crown of their Carpodetus serratus hosts. Most plants established more than 20 years ago, and the current potential for recruitment of new individuals into the population is severely limited by possum browsing and the senescent nature of the mistletoe population. Intensive management of host crowns and possum populations will be necessary to ensure the long-term viability of mistletoe at the study site.     

Relationships Among Families of Diprotodontia (Marsupialia) and the Phylogenetic Position of the Autapomorphic Honey Possum (Tarsipes rostratus)

The Australasian marsupial order Diprotodontia includes ten extant families that are grouped into the suborders Vombatiformes (koalas and wombats), Macropodiformes (kangaroos and allies), and Phalangeriformes (possums and gliders). We investigated interfamilial relationships using mitochondrial 12S rRNA, valine tRNA, and 16S rRNA gene sequences. Our results support the monophyly of both Vombatiformes and Macropodiformes, but not Phalangeriformes. Among possums and gliders, there was strong support for a petauroid clade that includes Pseudocheiridae (ringtail possums), Petauridae (sugar glider, striped possums), Acrobatidae (feathertail possums), and the monotypic family Tarsipedidae, which is represented by the highly specialized and autapomorphic honey possum (Tarsipes rostratus). Other prior hypotheses for the phylogenetic placement of the honey possum were rejected by statistical tests. The inclusion of the honey possum within Petauroidea suggests that derived ultrastructural features of Tarsipes' spermatozoa evolved independently in Tarsipes versus polyprotodont Australasian marsupials.     

The relationship between possum density and browse damage on kamahi in New Zealand forests

To minimize the impacts of introduced pests and to justify and prioritize pest control, managers need to know the relationship between pest density and damage. This relationship can be difficult to quantify because pest impacts can be highly variable. In New Zealand, introduced brushtail possums (Trichosurus vulpecula) browse a wide range of native forest species. However, possum browse is extremely patchy making it difficult to quantify the relationship between density and damage, meaning the benefits of reducing possum densities are poorly understood. We quantified patterns of possum browse on kamahi (Weinmannia racemosa), a common canopy tree species, at 21 forest sites that were repeat‐measured over an 8‐year period in the North Island, New Zealand, during which time possum densities fluctuated widely. We fitted a multilevel statistical model in order to quantify the relationship between possum density and browse damage while simultaneously quantifying how browse varied among trees, sites and years. Higher possum densities were associated with greater browse damage, but browse was also patchily distributed among trees at the same site, and among sites and years for a given possum density. This heterogeneity meant there was no simple density damage relationship, with the relationship differing from tree to tree and among sites and years. Our results show that at most sites reductions in possum density would have little benefit in reducing the probability of heavy browse on kamahi trees, but at a few sites there would be substantial benefits. This approach provides insights into the pattern and potential causes of variability in possum impacts, and a quantitative basis for prioritizing sites for possum control.     

Present Government policy on pest control

Abstract

The staff of Environment Waikato are to be congratulated for their initiative in setting up this important meeting. More public discussion on the pest problem in New Zealand is needed, although my mailbag suggests that there is already a keen awareness of the daunting problem that we face.

The possum is most often cast in the villain's role, but we should not ignore the rats, rabbits, deer, feral sheep, pigs, cats, mustelids, wasps, and feral cattle that are also pests in some ecosystems.     

High variability in the MHC class II DA beta chain of the brushtail possum (<Emphasis Type="Italic">Trichosurus vulpecula</Emphasis>)

The diversity of class II major histocompatibility complex (MHC) loci was investigated in the brushtail possum, an important marsupial pest species in New Zealand. Immunocontraception, a form of fertility control that generates an autoimmune response, is being developed as a population control method for the possum. Because the immune response is partly under genetic control, an understanding of immunogenetics in possum will be crucial to the development of immunocontraceptive vaccines. MHC molecules are critical in the vertebrate immune response. Class II MHC molecules bind and present exogenously derived peptides to T lymphocytes and may be important in the presentation of immunocontraceptives. We used polymerase chain reaction primers designed to amplify the peptide binding region of possum class II MHC genes to isolate sequences from 49 animals. We have previously described 19 novel alleles from the DAB locus in the possum (Holland et al., Immunogenetics 60:449–460, 2008). Here, we report on another 11 novel alleles isolated from possum DAB, making this the most diverse marsupial locus described so far. This high level of diversity indicates that DAB is an important MHC locus in the possum and will need to be taken into account in the design of immunocontraceptive vaccines.     

Resources at the landscape scale influence possum abundance

Abstract There is increasing realization that human perceptions of a ‘patchy’ environment do not necessarily relate to that of a particular organism and ecological characteristics of land adjacent to patches of natural vegetation (‘remnants’) may influence the ecological processes within. We investigated how the distribution and abundance of resources within and surrounding 39 small remnants located within the city of Melbourne, Australia, influenced the abundance of common brushtail possums Trichosurus vulpecula (Phalangeridae) and common ringtail possums Pseudocheirus peregrinus (Pseudocheiridae), two primarily folivorous marsupials. Bayesian modelling techniques were used to relate the abundance of both possum species to estimates of den and food availability both within the remnant and within a 100‐m buffer strip extending out into adjacent residential development. We found that both brushtail and ringtail possum abundance within remnants increased with the density of potential den sites within the remnant and food availability within the surrounding landscape. Ringtail possum density within the remnant further increased with den availability within the surrounding landscape. Our results lend support to the hypothesis that, in highly modified landscapes, processes originating outside a remnant vegetation patch may influence the abundance of species within.     

Browse on mahoe and kamahi leaf-fall as a trigger for possum control

Introduced brushtail possums (Trichosurus vulpecula) are controlled over large parts of New Zealand to protect canopy trees. The condition of canopy trees is one of the cues used to trigger possum control, but selecting an indicator of canopy tree condition is difficult because many factors unrelated to possum browsing can affect canopy condition, and indices based on canopy scoring may not always quickly detect real changes in possum herbivory. We therefore investigated the usefulness of the percentage of fallen mahoe (Melicytus ramiflorus) and kamahi (Weinmannia racemosa) leaves browsed by possums (‘fallen-leaf browse’ or ‘FLB’) as a trigger for control aimed at protecting these tree species. We collected leaves falling from kamahi and mahoe trees every two months for two years at two study areas in the central North Island, one with initially high possum abundance (Oriuwaka) and the other with low possum abundance (Otupaka). We classified each of 92 384 leaves as either not-browsed, possum-browsed or insect-browsed. There was a strong and similar seasonal pattern in the mean number of fallen leaves per tree for both mahoe and kamahi at both study areas; fewest leaves fell in winter, and the most leaves fell in spring and early summer. Mahoe and kamahi FLB exhibited a similar seasonal pattern at both areas, being lowest in winter and highest in spring and early summer. FLB for both mahoe and kamahi declined following control of possums to low densities at Oriuwaka. The proportion of fallen mahoe and kamahi leaves browsed by possums was small compared with those browsed by insects or not browsed. We show that spring/early summer (i.e. September–December) is the best period for sampling FLB and that the mean FLB can be estimated with a CV of 20% if one trap is randomly placed under the canopy of each of 24 randomly located trees. However, CVs were much larger in other seasons and when possum abundance was low. We consider FLB to be a sensible trigger for initiating possum control when the objective of control is to protect canopy trees, but further work is needed to determine the relationships between possum abundance, FLB, canopy condition, and key tree demographic rates.     

Cephalic morphology of the honey possum,Tarsipes rostratus (Marsupialia: Tarsipedidae); an obligate nectarivore

The sharply tapering skull of the honey possum is delicately constructed and has only a few, minute teeth; its mandible is reduced to a thin, flexible rod. The mandibular fossa has been diplaced caudally to the caudomedial corner of the squamosal. Head skeletons of the feathertail glider and western pygmy-possum, omnivores that are closely related to the honey possum, bear greater resemblance to the distantly related carnivorous fat-tailed dunnart than to the honey possum. Selected muscles associated with the jaws, hyoid, and tongue of these four mouse-sized (9–22 g) marsupials are described for the first time. The honey possum is characterized by a greatly reduced temporalis that is almost completely hidden by the eye. Its digastric consists of a single belly that inserts onto the caudal margin of the mylohyoid. The lateral pterygoid is relatively long as it extends caudally to insert onto the elongated mandible. The stylohyoid originates high up on the caudal surface of the tympanic bulla; it curves around the caudal and ventral surfaces of the bulla to reach the basihyoid. The insertion of the genioglossus is restricted to the caudal quarter of the tongue. Homologous muscles of the feathertail glider and western pygmy-possum are more similar to those of the fat-tailed dunnart. In addition to the very different musculoskeletal system, the honey possum has an unusual tongue that tapers to a fine point. © 1995 Wiley-Liss, Inc.     

Predicting Summer Site Occupancy for an Invasive Species,the Common Brushtail Possum (Trichosurus vulpecula), in an Urban Environment

Invasive species are often favoured in fragmented, highly-modified, human-dominated landscapes such as urban areas. Because successful invasive urban adapters can occupy habitat that is quite different from that in their original range, effective management programmes for invasive species in urban areas require an understanding of distribution, habitat and resource requirements at a local scale that is tailored to the fine-scale heterogeneity typical of urban landscapes. The common brushtail possum (Trichosurus vulpecula) is one of New Zealand’s most destructive invasive pest species. As brushtail possums traditionally occupy forest habitat, control in New Zealand has focussed on rural and forest habitats, and forest fragments in cities. However, as successful urban adapters, possums may be occupying a wider range of habitats. Here we use site occupancy methods to determine the distribution of brushtail possums across five distinguishable urban habitat types during summer, which is when possums have the greatest impacts on breeding birds. We collected data on possum presence/absence and habitat characteristics, including possible sources of supplementary food (fruit trees, vegetable gardens, compost heaps), and the availability of forest fragments from 150 survey locations. Predictive distribution models constructed using the programme PRESENCE revealed that while occupancy rates were highest in forest fragments, possums were still present across a large proportion of residential habitat with occupancy decreasing as housing density increased and green cover decreased. The presence of supplementary food sources was important in predicting possum occupancy, which may reflect the high nutritional value of these food types. Additionally, occupancy decreased as the proportion of forest fragment decreased, indicating the importance of forest fragments in determining possum distribution. Control operations to protect native birds from possum predation in cities should include well-vegetated residential areas; these modified habitats not only support possums but provide a source for reinvasion of fragments.     

Effect of dietary terpenes on glucuronic acid excretion and ascorbic acid turnover in the brushtail possum (Trichosurus vulpecula)

1. Glucuronic acid was excreted in the urine of the brushtail possum, Trichosurus vulpecula, in response to dietary terpenes (essential oils found in Eucalyptus foliage). 2. The relationship between urinary glucuronic acid and the estimated terpene content of the diet was not equimolar, but varied from 1-17%, with levels of 5-39% found in animals maintained on terpenes or eucalypt leaves for several weeks. 3. Blood levels of ascorbate and the turnover rate of ascorbate were increased in the brushtail possum in response to dietary terpenes. 4. The presence of an active glucuronic acid pathway and associated glucose-ascorbate-glucose cycle was postulated for the brushtail possum and the other arboreal marsupials, Pseudocheirus peregrinus (common ringtail possum) and Petauroides volans (greater glider). 5. By means of these pathways these animals may utilise the high ascorbate content of Eucalyptus leaves to conserve glucose required for synthesis of glucuronic acid used for conjugation of dietary terpenes.     

Expression in Escherichia coli and immunological characterization of three zona pellucida proteins (ZP1, ZP2, and ZP3) from a marsupial,the brushtail possum (Trichosurus vulpecula)

The brushtail possum (Trichosurus vulpecula) zona pellucida (ZP) is composed of three major glycoproteins, designated ZP1, ZP2, and ZP3 based on their size and homology with eutherian ZP proteins. These proteins are candidate antigens for the development of an immunocontraceptive vaccine to control the fertility of the brushtail possum in New Zealand, where it is an introduced pest. In order to further their immunological and functional characterization, recombinant possum ZP proteins were produced in Escherichia coli (E. coli) strain JM109, M15, SG13009, or BL21 codon plus. Each of the proteins produced possessed a N-terminal six histidine tag (His)(6) to facilitate purification and consisted of amino acid (aa) residues 18-471 of possum ZP1, aa residues 40-311 of ZP2 (ZP2-N), aa residues 305-634 of ZP2 (ZP2-C), and aa residues 23-342 of ZP3. Immunoblot using anti-RGS(His)(4) antibodies and polyclonal rabbit anti-porcine ZP antibodies detected major bands at 54 kDa for ZP1, 32 kDa for ZP2-N, 39 kDa for ZP2-C, and 40 kDa for ZP3. Immunization of male and female rabbits with ZP2-N, ZP2-C, and ZP3 purified on Ni-NTA resin under denaturing conditions generated antibodies reactive with recombinant ZP proteins on Western blot and with native ZP proteins in possum ovarian sections using immunofluorescence. Antibodies generated against ZP1 in the same way were reactive with recombinant ZP proteins on Western blot only. The recombinant possum ZP proteins and specific antibodies produced in this study give an indication of the antigenic relationship of the possum ZP proteins and are vital tools for future studies of sperm-ZP binding in marsupials and for the evaluation of ZP-based contraceptive vaccines in possums and other marsupials.     

Spatial variation in impacts of brushtail possums on two Loranthaceous mistletoe species

Browsing by introduced brushtail possums is linked to major declines in mistletoe abundance in New Zealand, yet in some areas mistletoes persist, apparently unaffected by the presence of possums. To determine the cause of this spatial variation in impact I investigated the abundance and condition (crown dieback and extent of possum browse) of two mistletoes (Alepis flavida, Peraxilla tetrapetala) and abundance and diet of possums in two mountain beech (Nothofagus solandri var. cliffortioides) forests in the central-eastern South Island of New Zealand. Mistletoe is common and there are long-established uncontrolled possum populations in both forests. Mistletoes were abundant (216?1359 per hectare) and important in possum diet (41?59% of total diet), but possum density was low (c. 2 per hectare) in both areas. Possum impacts were slight with low browse frequencies and intensities over much of the study sites. However, impacts were significantly greater at a forest margin, where possum abundance was highest, and at a high-altitude site where mistletoe density was lowest. Mistletoe crown dieback was inversely proportional to intensity of possum browsing. These results suggest that the persistence of abundant mistletoe populations at these sites is due to mistletoe productivity matching or exceeding consumption by possums in these forests of low possum-carrying capacity, rather than low possum preference for the local mistletoe populations.     

Identification of novel major histocompatibility complex class I sequences in a marsupial,the brushtail possum (<Emphasis Type="Italic">Trichosurus vulpecula</Emphasis>)

The major histocompatibility complex (MHC) is an essential part of the vertebrate immune response. MHC genes may be classified as classical, non-classical or non-functional pseudogenes. We have investigated the diversity of class I MHC genes in the brushtail possum, a marsupial native to Australia and an introduced pest in New Zealand. The MHC of marsupials is poorly characterised compared to eutherian mammal species. Comparisons between marsupials and eutherians may enhance understanding of the evolution and functions of this important genetic region. We found a high level of diversity in possum class I MHC genes. Twenty novel sequences were identified using polymerase chain reaction (PCR) primers designed from existing marsupial class I MHC genes. Eleven of these sequences shared a high level of homology with the only previously identified possum MHC class I gene TrvuUB and appear to be alleles at a single locus. Another seven sequences are also similar to TrvuUB but have frame-shift mutations or stop codons early in their sequence, suggesting they are non-functional alleles of a pseudogene locus. The remaining sequences are highly divergent from other possum sequences and clusters with American marsupials in phylogenetic analysis, indicating they may have changed little since the separation of Australian and American marsupials.     

Clinical,Microbiological and Pathological Findings of Mycobacterium ulcerans Infection in Three Australian Possum Species

Background

Buruli ulcer (BU) is a skin disease caused by Mycobacterium ulcerans, with endemicity predominantly in sub-Saharan Africa and south-eastern Australia. The mode of transmission and the environmental reservoir(s) of the bacterium and remain elusive. Real-time PCR investigations have detected M. ulcerans DNA in a variety of Australian environmental samples, including the faeces of native possums with and without clinical evidence of infection. This report seeks to expand on previously published findings by the authors'' investigative group with regards to clinical and subclinical disease in selected wild possum species in BU-endemic areas of Victoria, Australia.

Methodology/Principal Findings

Twenty-seven clinical cases of M. ulcerans infection in free-ranging possums from southeastern Australia were identified retrospectively and prospectively between 1998–2011. Common ringtail possums (Pseudocheirus peregrinus), a common brushtail possum (Trichosurus vulpecula) and a mountain brushtail possum (Trichosurus cunninghami) were included in the clinically affected cohort. Most clinically apparent cases were adults with solitary or multiple ulcerative cutaneous lesions, generally confined to the face, limbs and/or tail. The disease was minor and self-limiting in the case of both Trichosurus spp. possums. In contrast, many of the common ringtail possums had cutaneous disease involving disparate anatomical sites, and in four cases there was evidence of systemic disease at post mortem examination. Where tested using real-time PCR targeted at IS2404, animals typically had significant levels of M. ulcerans DNA throughout the gut and/or faeces. A further 12 possums without cutaneous lesions were found to have PCR-positive gut contents and/or faeces (subclinical cases), and in one of these the organism was cultured from liver tissue. Comparisons were made between clinically and subclinically affected possums, and 61 PCR-negative, non-affected individuals, with regards to disease category and the categorical variables of species (common ringtail possums v others) and sex. Animals with clinical lesions were significantly more likely to be male common ringtail possums.

Conclusions/Significance

There is significant disease burden in common ringtail possums (especially males) in some areas of Victoria endemic for M. ulcerans disease. The natural history of the disease generally remains unknown, however it appears that some mildly affected common brushtail and mountain brushtail possums can spontaneously overcome the infection, whereas some severely affected animals, especially common ringtail possums, may become systemically, and potentially fatally affected. Subclinical gut carriage of M. ulcerans DNA in possums is quite common and in some common brushtail and mountain brushtail possums this is transient. Further work is required to determine whether M. ulcerans infection poses a potential threat to possum populations, and whether these animals are acting as environmental reservoirs in certain geographical areas.     

Long‐term response of temperate canopy trees to removal of browsing from an invasive arboreal herbivore in New Zealand

Defoliation of forest tree canopies by herbivores and other agents, leading to tree mortality and reduced productivity, threatens the ecological stability of forests globally. This study shows that long‐term control of a mammalian arboreal folivore (brushtail possums; Trichosurus vulpecula Phalangeridae) reduces crown dieback and increases foliage cover in browsing‐damaged canopy trees. We monitored indices of possum density, possum browsing, tree foliage cover and crown dieback for 20 years following initiation of possum control in 1994 that repeatedly reduced possum densities to near zero every 5–6 years and kept the population below 35% of pre‐control levels over the entire period. Observable possum browsing was recorded on 20–49% of individuals of three palatable tree species at the time of first control. Those percentages fell to zero after control and never exceeded 2–10% for individual species over the next 19 years. We recorded significant increases in foliage cover attributable to recovery from defoliation by possums for all three species during the first 10 years. Large increases in foliage cover occurred on individuals that were heavily browsed in 1994 (mean increases: 36–89%), but mean population increases were modest (3–19%) because only 10–19% of trees were initially heavily browsed. Twenty‐year mortality rates were similar for plants with, or without, initial possum browsing, indicating no residual impact of pre‐control browsing on tree mortality. Times for full recovery of crown foliage cover varied from 10 years for the youngest trees and faster growing species to more than 20 years for mature individuals of the slowest growing species.