Exploring patterns of female house mouse spatial organisation among outbreaking and stable populations.

The size and distribution of home ranges reflect how individuals within a population use, defend, and share space and resources, and may thus be an important predictor of population-level dynamics. Eruptive species, such as the house mouse in Australian grain-growing regions, are an ideal species in which to investigate variations in space use and home range overlap between stable and outbreaking populations. In this study, we use spatially explicit capture-recapture models to explore if space use and home range overlap among female mice could serve as indicators of changes in population density leading into summer. Additionally, we assess the sensitivity of space use and home range estimates to reduced recapture rates. Our analysis did not reveal variations in the spring spatial organisation of female mice based on existing capture-mark-recapture data. However, our study highlights the need to balance monitoring efforts within regions, emphasising the importance of exploring studies that can improve spatial recaptures by optimising trapping efforts. This is particularly important in Australian agricultural systems, where varying farm management practices may drive differences in population dynamics.

Developing fertility control for rodents: a framework for researchers and practitioners.

Fertility control is often heralded as a humane and effective technique for management of overabundant wildlife, including rodents. The intention is to reduce the use of lethal and inhumane methods, increase farm productivity and food security as well as reduce disease transmission, particularly of zoonoses. We developed a framework to guide researchers and stakeholders planning to assess the effectiveness of a potential contraceptive agent for a particular species. Our guidelines describe the overarching research questions which must be sequentially addressed to ensure adequate data are collected so that a contraceptive can be registered for use in broad-scale rodent management. The framework indicates that studies should be undertaken iteratively and, at times, in parallel, with initial research being conducted on (1) laboratory-based captive assessments of contraceptive effects in individuals; (2) simulation of contraceptive delivery using bait markers and/or surgical sterilization of different proportions of a field-based or enclosure population to determine how population dynamics are affected; (3) development of mathematical models which predict the outcomes of different fertility control scenarios; and (4) implementation of large-scale, replicated trials to validate contraceptive efficacy under various management-scale field situations. In some circumstances, fertility control may be most effective when integrated with other methods (e.g. some culling). Assessment of non-target effects, direct and indirect, and the environmental fate of the contraceptive must also be determined. Developing fertility control for a species is a resource-intensive commitment but will likely be less costly than the ongoing environmental and economic impacts by rodents and rodenticides in many contexts.

Effects of harvesting and stubble management on abundance of pest rodents (Mus musculus) in a conservation agriculture system.

BACKGROUND: The shift to more environmentally sensitive agricultural practices over the last several decades has changed farmland landscapes worldwide. Changes including no-till and retaining high biomass mulch has been coincident with an increase in rodent pests in South Africa, India, South America and Europe, indicating a possible conflict between conservation agriculture (CA) and rodent pest management. Research on effects of various crop management practices associated with CA on pest rodent population dynamics is needed to anticipate and develop CA-relevant management strategies. RESULTS: During the Australian 2020-2021 mouse plague, farmers used postharvest stubble management practices, including flattening and/or cutting, to reduce stubble cover in paddocks to lessen habitat suitability for pest house mice. We used this opportunity to assess the effects of both harvest and stubble management on the movement and abundance of mice in paddocks using mouse trapping and radio tracking. We found that most tracked mice remained resident in paddocks throughout harvest, and that mouse population abundance was generally unaffected by stubble management. CONCLUSION: Recent conversions to CA practices have changed how pest house mice use cropped land. Management practices that reduce postharvest habitat complexity do not appear to reduce the attractiveness of paddocks to mice, and further research into new management strategies in addition to toxic bait use is required as part of an integrated pest management approach. © 2023 The Authors. Pest Management Science published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.

Acute oral toxicity of zinc phosphide: an assessment for wild house mice (Mus musculus).

Irregular plagues of house mice, Mus musculus, incur major economic impacts on agricultural production in Australia. The efficacy of zinc phosphide (ZnP), the only registered broadacre control agent for mice, is reported as increasingly variable. Have mice become less sensitive over time or are they taking a sub-lethal dose and developing aversion? In this laboratory study, the sensitivity of mice (wild caught; outbred laboratory strain) was assessed using oral gavage of a range of ZnP concentrations. The estimated LD50 values (72-79 mg ZnP/kg body weight) were similar for each mouse group but are significantly higher than previously reported. The willingness of mice to consume ZnP-coated grains was determined. ZnP-coated grains (50 g ZnP/kg grain) presented in the absence of alternative food were consumed and 94% of wild mice died. Mice provided with alternative food and ZnP-coated wheat grains (either 25 or 50 g ZnP/kg grain) consumed toxic and non-toxic grains, and mortality was lower (33-55%). If a sublethal amount of ZnP-coated grain was consumed, aversion occurred, mostly when alternative food was present. The sensitivity of wild house mice to ZnP in Australia is significantly lower than previously assumed. Under laboratory conditions, ZnP-coated grains coated with a new higher dose (50 g ZnP/kg grain) were readily consumed. Consumption of toxic grain occurred when alternative food was available but was decreased. Our unambiguous findings for house mice indicate a re-assessment of the ZnP loading for baits used for control of many rodents around the world may be warranted.

Sensitivity of the stripe-faced dunnart, Sminthopsis macroura (Gould 1845), to the insecticide, fipronil; implications for pesticide risk assessments in Australia.

A lack of toxicity data quantifying responses of Australian native mammals to agricultural pesticides prompted an investigation into the sensitivity of the stripe-faced dunnart, Sminthopsis macroura (Gould 1845) to the insecticide, fipronil (5-amino-3-cyano-1-(2,6-dichloro-4-trifluoromethylphenyl)-4-trifluoromethylsulfinyl pyrazole, CAS No. 120068-37-3). Using the Up-And-Down method for determining acute oral toxicity in mammals (OECD) median lethal dose estimates of 990 mg kg-1 (95% confidence interval (CI) = 580.7-4770.0 mg kg-1) and 270.4 mg kg-1 (95% CI = 0.0->20,000.0 mg kg-1) were resolved for male and female S. macroura, respectively. The difference between median lethal dose estimates for males and females may have been influenced by the older ages of two female dunnarts. Consequently, further modelling of female responses to fipronil doses used the following assumptions: (a) death at 2000 mg kg-1, (b) survival at 500 mg kg-1 and (c) a differential response (both survival and death) at 990 mg kg-1. This modelling revealed median lethal dose estimates for female S. macroura of 669.1 mg kg-1 (95% CI = 550-990 mg kg-1; assuming death at 990 mg kg-1) and 990 mg kg-1 (95% CI = 544.7-1470 mg kg-1; assuming survival at 990 mg kg-1). These median lethal dose estimates are 3-10-fold higher than available LD50 values of 94 mg kg-1 for a similarly sized eutherian mammal, Mus musculus (L. 1758) and 97 mg kg-1 for Rattus norvegicus (Birkenhout 1769). Implications for pesticide risk assessments in Australia are discussed.

Reproductive responses of rice field rats (Rattus argentiventer) following treatment with the contraceptive hormones, quinestrol and levonorgestrol.

The rice field rat, Rattus argentiventer, is a significant pest of rice in Southeast Asia. Fertility control methods have the potential to provide safe and effective alternatives to control methods that often include indiscriminate use of rodenticides or electric barriers. The aim of this laboratory study was to assess uptake of bait coated with different concentrations of the contraceptive hormones, quinestrol (E) and levonorgestrel (P), delivered alone and in combination (i.e. EP-1) and determine the short-term effects on reproductive parameters of adult male and female R. argentiventer. In Experiment 1, 2 concentrations of E, P, and EP-1 (10, 20 ppm) were fed to groups of wild-caught rats for 7 days. In females, both E and EP-1 induced uterine edema. In males, EP-1 reduced epididymis and seminal vesicle weights and lowered sperm motility. However, these responses were inconsistent due to low bait acceptance, especially with increasing concentrations. In Experiment 2, EP-1 (0, 20, 50, 100 ppm) was administered by oral gavage daily for 7 days to male R. argentiventer. There were significant reductions in epididymal and seminal vesicle weights for all oral doses of EP-1, in sperm counts for the 50 ppm dose, and in sperm motility for the 20 and 50 ppm doses compared to the control group. To select the optimum dose of EP-1, we must address the poor acceptance of contraceptive-coated baits by rice field rats. Further research is required to improve the palatability of EP-1 and to test its uptake under field conditions.

The status of fertility control for rodents-recent achievements and future directions.

Management of overabundant rodents at a landscape scale is complex but often required to sustainably reduce rodent abundance below damage thresholds. Current conventional techniques such as poisoning are not species specific, with some approaches becoming increasingly unacceptable to the general public. Fertility control, first proposed for vertebrate pest management over 5 decades ago, has gained public acceptance because it is perceived as a potentially more species-specific and humane approach compared with many lethal methods. An ideal fertility control agent needs to induce infertility across one or more breeding seasons, be easily delivered to an appropriate proportion of the population, be species specific with minimal side-effects (behavioral or social structure changes), and be environmentally benign and cost effective. To date, effective fertility control of rodents has not been demonstrated at landscape scales and very few products have achieved registration. Reproductive targets for fertility control include disrupting the hormonal feedback associated with the hypothalamic-pituitary-gonadal axis, gonad function, fertilization, and/or early implantation. We review progress on the oral delivery of various agents for which laboratory studies have demonstrated efficacy in females and/or males and synthesize progress with the development and/or use of synthetic steroids, plant extracts, ovarian specific peptides, and immunocontraceptive vaccines. There are promising results for field application of synthetic steroids (levonorgestrel, quinestrol), chemosterilants (4-vinylcyclohexene diepoxide), and some plant extracts (triptolide). For most fertility control agents, more research is essential to enable their efficient and cost-effective delivery such that rodent impacts at a population level are mitigated and food security is improved.

Effects of background food on alternative grain uptake and zinc phosphide efficacy in wild house mice.

BACKGROUND: House mice (Mus musculus) cause significant, ongoing losses to grain crops in Australia, particularly during mouse plagues. Zinc phosphide (ZnP) coated grain is used for control, but with variable success. In a laboratory setting, we tested if mice would (i) switch from consumption of one grain type to another when presented with an alternative and (ii) consume ZnP-treated grains when presented as a choice with a different grain. RESULTS: Mice readily switched from their background grain to an alternative grain, preferring cereals (wheat or barley) over lentils. Mice readily consumed ZnP-coated barley grains. Their mortality rate was significantly higher (86%, n = 30) in the presence of a less-favoured grain (lentils) compared to their mortality rate (47%, n = 29; 53%, n = 30) in the presence of a more-favoured grain (wheat and barley, respectively). Mice died between 4 and 112 h (median = 18 h) after consuming one or more toxic grains. Independent analysis of ZnP-coated grains showed variable toxin loading indicating that consumption of a single grain would not guarantee intake of a lethal dose. There was also a strong and rapid behavioural aversion if mice did not consume a lethal dose on the first night. CONCLUSIONS: The registered dose rate of 25 g of ZnP/kg wheat (~1 mg of ZnP/grain) in Australia needs to be re-evaluated to determine what factors may be contributing to variation in efficacy. Further field research is also required to understand the complex association between ZnP dose, and quantity and quality of background food on efficacy of ZnP baits.

Efficacy of a combined insecticide-rodenticide product on ectoparasite and commensal rodent mortality.

BACKGROUND: Ectoparasites may transfer zoonotic pathogens from rodents to humans or livestock when rodents are managed with rodenticides. This could be minimized using a product combining a rodenticide with a delayed action and a systemic insecticide/acaricide that rapidly kills ectoparasites. Such a combination was tested in commensal pest rodent species to assess efficacy and timing of responses in rodents, and fleas and ticks feeding on them. Ticks or fleas attached to rats (Rattus norvegicus) and house mice (Mus musculus domesticus) were exposed to a product containing brodifacoum (50 ppm) and fipronil (40 ppm) for three days. RESULTS: 98-100% of fleas on treated rodents died within one to two days after first exposure, whereas >90% fleas survived on control rodents. The effect persisted for four or more days after bait uptake. Ticks started to succumb to the effect of the combination product within one day (mice) and within four days (rats) of first exposure, with all ticks dying by Day (D)8. Tick survival in control rodents was 90-100%. Rodent mortality began at D3 (rats) and D4 (mice) after first consumption of product and all were dead by D9 (rats) and D7 (mice). CONCLUSION: This product effectively killed ectoparasites and rodents. Flea mortality was swift and complete, generally within one day of exposure, whereas it took ticks up to four days to die, but before the rats and house mice died. The combination product might help to prevent ectoparasites migrating from dying rodents to another host. Field trials are warranted. © 2020 Society of Chemical Industry.

Induction of synchronous oestrus but not ovulation after pre-treatment with the GnRH agonist, Lucrin® Depot, in the tammar wallaby.

We investigated the capacity for pregnant mare serum gonadotrophin (PMSG) and human chorionic gonadotrophin (hCG) to induce synchronous oestrus and ovulation in the tammar wallaby (Notamacropus eugenii) after follicular suppression with Lucrin® Depot, a one-month GnRH agonist. On Day 0 pouch young were removed (RPY) to reactivate a normal oestrous cycle and then two groups were treated with vehicle (Control; n = 5 and Superovulation (SOvn); n = 5) and two groups were treated with 7.5 mg of Lucrin Depot (Lucrin; n = 6; Lucrin+SOvn, n = 6). On Day 20 RPY the SOvn and Lucrin+SOvn Group received 20 IU of PMSG, which was followed on Day 23 RPY with 500 IU of hCG. The Lucrin+SOvn females underwent a more synchronous oestrus with 5 of 6 mating on Day 26 RPY while the SOvn (n = 5/5) and Control (n = 5/5) Groups copulated over two days, between Day 25-27 RPY and Day 27-29 RPY respectively. Mating plugs were not detected in any of the females in the Lucrin Group by Day 31 RPY. Autopsy on Day 31 RPY confirmed all females in each treatment group had undergone a reactivated cycle as evidenced by the presence of a large corpus luteum (CL) in one ovary. At autopsy the females in the Lucrin+SOvn Group had highly stimulated reproductive tracts, and their ovaries contained many follicles >3 mm; 14 ± 2.1 and 15.3 ± 2.1 follicles >3 mm in the CL-bearing ovary and contralateral ovary respectively. Similarly, females in the SOvn Group had 11.4 ± 2.4 and 17.4 ± 1.9 follicles >3 mm in each respective ovary. Uterine flushing and ovarian histology confirmed that females in Lucrin+SOvn and SOvn Groups had not ovulated, but normal oocytes were present in the follicles. By comparison, the Control Group had ovulated with a single embryo being recovered from the uterus of 4 of 5 females. In contrast to all groups, females in the Lucrin Group showed follicular suppression (all follicles <1.5 mm) and an unstimulated reproductive tract. We conclude that a suppression plus stimulation regimen using Lucrin Depot followed by PMSG and hCG has the capacity to synchronise oestrus, and that 20 IU of PMSG stimulates the development of antral follicles >3 mm in both ovaries. However, a single 500 IU treatment of hCG on Day 23 RPY was not able to induce ovulation in the tammar wallaby.

Guiding Development of the Neonate: Lessons from Mammalia.

Significantly preterm and low-birthweight (LBW) babies have diminished lung and gut development, generally fail to thrive, have increased mortality and higher frequency of mature-onset disease. Mothers often cannot breastfeed, and babies receive either formula or pasteurized donor milk, which may further limit the baby's recovery. New approaches are required to manage the early stages of neonatal development. The tammar wallaby, an Australian marsupial, has a short gestation and a simple placenta, and gives birth to an altricial young equivalent to a final trimester human embryo. The neonate remains in the pouch and attached to the teat for 100 days postpartum. The mother slows growth of the young and progressively changes the composition of the milk to deliver signals for organ development, including the lung and gut. This closely resembles the relationship between the human fetus and delivery of placental and uterine bioactives. Datasets comprised of differentially expressed genes coding for secreted proteins in early lactation in the tammar mammary gland have been compared to databases produced from human placenta, amniotic fluid, colostrum and milk to identify human homologues for the putative signaling molecules for organ development. These data will be used to develop milk fortifiers for treatment of preterm and LBW babies in both the developed and the developing world.

Delayed return to estrus following treatment with the gonadotrophin-releasing hormone agonist, Lucrin® Depot, in the tammar wallaby.

Gonadotrophin-releasing hormone agonists that induce a reversible contraceptive state in several marsupials have the potential to be used to synchronize estrus. We used a model macropod, the tammar wallaby (Notamacropus eugenii), to investigate whether Lucrin Depot (AbbVie), a GnRH agonist microsphere preparation, could (1) inhibit follicular development and estrus in a cycle reactivated by removal of pouch young (RPY) and (2) facilitate a synchronous return to estrus. Our results show that females reactivated with bromocriptine and RPY in early seasonal quiescence (July 2015) were inhibited by Lucrin Depot (0.125-0.5 mg kg -1, n = 9) and unlike control females (n = 3), did not copulate before Day 32 RPY. During the next breeding season (February 2016), the return to estrus after RPY was not delayed in animals treated with Lucrin Depot (≤0.20 mg kg -1; n = 12), and copulation occurred in treated and control females within the expected natural period after RPY (Day 26-33 RPY). In the following breeding season (March 2017), estrus was delayed in animals treated with Lucrin Depot (1.25 mg kg -1) on either Day 0 (Group A, n = 6) or Day 10 (Group B, n = 6) after RPY compared to control females (n = 6). Estrus was detected in Group A between 39 and 66 days (55 ±â€¯4.8d) and in Group B between 43 and 71 days (55.2 ±â€¯3.9d) after RPY. In contrast, all control females underwent estrus and copulated as expected by Day 30 RPY. We conclude Lucrin Depot can inhibit ovarian follicular activity after RPY but as a standalone treatment does not result in a highly synchronous return to estrus in the tammar wallaby.

A bipedal mammalian model for spinal cord injury research: The tammar wallaby.

Background: Most animal studies of spinal cord injury are conducted in quadrupeds, usually rodents. It is unclear to what extent functional results from such studies can be translated to bipedal species such as humans because bipedal and quadrupedal locomotion involve very different patterns of spinal control of muscle coordination. Bipedalism requires upright trunk stability and coordinated postural muscle control; it has been suggested that peripheral sensory input is less important in humans than quadrupeds for recovery of locomotion following spinal injury. Methods: We used an Australian macropod marsupial, the tammar wallaby (Macropuseugenii), because tammars exhibit an upright trunk posture, human-like alternating hindlimb movement when swimming and bipedal over-ground locomotion. Regulation of their muscle movements is more similar to humans than quadrupeds. At different postnatal (P) days (P7-60) tammars received a complete mid-thoracic spinal cord transection. Morphological repair, as well as functional use of hind limbs, was studied up to the time of their pouch exit. Results: Growth of axons across the lesion restored supraspinal innervation in animals injured up to 3 weeks of age but not in animals injured after 6 weeks of age. At initial pouch exit (P180), the young injured at P7-21 were able to hop on their hind limbs similar to age-matched controls and to swim albeit with a different stroke. Those animals injured at P40-45 appeared to be incapable of normal use of hind limbs even while still in the pouch. Conclusions: Data indicate that the characteristic over-ground locomotion of tammars provides a model in which regrowth of supraspinal connections across the site of injury can be studied in a bipedal animal. Forelimb weight-bearing motion and peripheral sensory input appear not to compensate for lack of hindlimb control, as occurs in quadrupeds. Tammars may be a more appropriate model for studies of therapeutic interventions relevant to humans.

Testing hypotheses of developmental constraints on mammalian brain partition evolution, using marsupials.

There is considerable debate about whether the partition volumes of the mammalian brain (e.g. cerebrum, cerebellum) evolve according to functional selection, or whether developmental constraints of conserved neurogenetic scheduling cause predictable partition scaling with brain size. Here we provide the first investigation of developmental constraints on partition volume growth, derived from contrast-enhanced micro-computed tomography of hydrogel-stabilized brains from three marsupial species. ANCOVAs of partition vs. brain volume scaling, as well as growth curve comparisons, do not support several hypotheses consistent with developmental constraints: brain partition growth significantly differs between species, or between developing vs. adult marsupials. Partition growth appears independent of adult brain volume, with no discernable growth spurts/lags relatable to internal structural change. Rather, adult proportion differences appear to arise through growth rate/duration heterochrony. Substantial phylogenetic signal in adult brain partitions scaling with brain volume also counters expectations of development-mediated partition scaling conservatism. However, the scaling of olfactory bulb growth is markedly irregular, consistent with suggestions that it is less constrained. The very regular partition growth curves suggest intraspecific developmental rigidity. We speculate that a rigid, possibly neuromer-model-like early molecular program might be responsible both for regular growth curves within species and impressions of a link between neurogenesis and partition evolution.

The tammar wallaby: A marsupial model to examine the timed delivery and role of bioactives in milk.

It is now clear that milk has multiple functions; it provides the most appropriate nutrition for growth of the newborn, it delivers a range of bioactives with the potential to stimulate development of the young, it has the capacity to remodel the mammary gland (stimulate growth or signal cell death) and finally milk can provide protection from infection and inflammation when the mammary gland is susceptible to these challenges. There is increasing evidence to support studies using an Australian marsupial, the tammar wallaby (Macropus eugenii), as an interesting and unique model to study milk bioactives. Reproduction in the tammar wallaby is characterized by a short gestation, birth of immature young and a long lactation. All the major milk constituents change substantially and progressively during lactation and these changes have been shown to regulate growth and development of the tammar pouch young and to have roles in mammary gland biology. This review will focus on recent reports examining the control of lactation in the tammar wallaby and the timed delivery of milk bioactivity.

Marsupial tammar wallaby delivers milk bioactives to altricial pouch young to support lung development.

Our research is exploiting the marsupial as a model to understand the signals required for lung development. Marsupials have a unique reproductive strategy, the mother gives birth to altricial neonate with an immature lung and the changes in milk composition during lactation in marsupials appears to provide bioactives that can regulate diverse aspects of lung development, including branching morphogenesis, cell proliferation and cell differentiation. These effects are seen with milk collected between 25 and 100days postpartum. To better understand the temporal effects of milk composition on postnatal lung development we used a cross-fostering technique to restrict the tammar pouch young to milk composition not extending beyond day 25 for 45days of its early postnatal life. These particular time points were selected as our previous study showed that milk protein collected prior to ~day 25 had no developmental effect on mouse embryonic lungs in culture. The comparative analysis of the foster group and control young at day 45 postpartum demonstrated that foster pouch young had significantly reduced lung size. The lungs in fostered young were comprised of large intermediate tissue, had a reduced size of airway lumen and a higher percentage of parenchymal tissue. In addition, expression of marker genes for lung development (BMP4, WNT11, AQP-4, HOPX and SPB) were significantly reduced in lungs from fostered young. Further, to identify the potential bioactive expressed by mammary gland that may have developmental effect on pouch young lungs, we performed proteomics analysis on tammar milk through mass-spectrometry and listed the potential bioactives (PDGF, IGFBP5, IGFBPL1 and EGFL6) secreted in milk that may be involved in regulating pouch young lung development. The data suggest that postnatal lung development in the tammar young is most likely regulated by maternal signalling factors supplied through milk.

Mammalian development does not recapitulate suspected key transformations in the evolutionary detachment of the mammalian middle ear.

The ectotympanic, malleus and incus of the developing mammalian middle ear (ME) are initially attached to the dentary via Meckel's cartilage, betraying their origins from the primary jaw joint of land vertebrates. This recapitulation has prompted mostly unquantified suggestions that several suspected--but similarly unquantified--key evolutionary transformations leading to the mammalian ME are recapitulated in development, through negative allometry and posterior/medial displacement of ME bones relative to the jaw joint. Here we show, using µCT reconstructions, that neither allometric nor topological change is quantifiable in the pre-detachment ME development of six marsupials and two monotremes. Also, differential ME positioning in the two monotreme species is not recapitulated. This challenges the developmental prerequisites of widely cited evolutionary scenarios of definitive mammalian middle ear (DMME) evolution, highlighting the requirement for further fossil evidence to test these hypotheses. Possible association between rear molar eruption, full ME ossification and ME detachment in marsupials suggests functional divergence between dentary and ME as a trigger for developmental, and possibly also evolutionary, ME detachment. The stable positioning of the dentary and ME supports suggestions that a 'partial mammalian middle ear' as found in many mammaliaforms--probably with a cartilaginous Meckel's cartilage--represents the only developmentally plausible evolutionary DMME precursor.

Role of marsupial tammar wallaby milk in lung maturation of pouch young.

BACKGROUND: Marsupials such as the tammar wallaby (M.Eugenii) have a short gestation (29.3 days) and at birth the altricial young resembles a fetus, and the major development occurs postnatally while the young remains in the mother's pouch. The essential functional factors for the maturation of the neonate are provided by the milk which changes in composition progressively throughout lactation (300 days). Morphologically the lungs of tammar pouch young are immature at birth and the majority of their development occurs during the first 100 days of lactation. RESULTS: In this study mouse embryonic lungs (E-12) were cultured in media with tammar skim milk collected at key time points of lactation to identify factors involved in regulating postnatal lung maturation. Remarkably the embryonic lungs showed increased branching morphogenesis and this effect was restricted to milk collected at specific time points between approximately day 40 to 100 lactation. Further analysis to assess lung development showed a significant increase in the expression of marker genes Sp-C, Sp-B, Wnt-7b, BMP4 and Id2 in lung cultures incubated with milk collected at day 60. Similarly, day 60 milk specifically stimulated proliferation and elongation of lung mesenchymal cells that invaded matrigel. In addition, this milk stimulated proliferation of lung epithelium cells on matrigel, and the cells formed 3-dimensional acini with an extended lumen. CONCLUSIONS: This study has clearly demonstrated that tammar wallaby milk collected at specific times in early lactation contains bioactives that may have a significant role in lung maturation of pouch young.