Reproductive biology of the Red-necked wallaby (Macropus rufogriseus banksianus) and Bennett's wallaby (M. r. rufogriseus) in captivity

Bennett's wallaby ( Macropus r. rufogriseus ) of Tasmania give birth from late January to early August in marked contrast to the Red-necked wallaby ( M. r. banksianus ) of mainland south-eastern Australia which produced young in all months. Within the breeding season however, the lengths of the oestrous cycle and gestation period are similar in the two forms and did not differ by more than 0.5 days. The gestation period of about 30 days extended to almost the length of the oestrous cycle of approximately 33 days. Birth was closely followed by mating which normally resulted in fertilization and subsequent embryonic diapause. Renewed blastocyst development was initiated by removal or loss of a pouch young and birth followed about 27 days later.
Unlike other macropodids with a similar breeding pattern, birth, as a result of renewed blastocyst development near the end of a large young's pouch life, did not occur within a day or two of the permanent emergence of the young, but followed 16 to 29 days later. In M. r. rufogriseus , young that left the pouch permanently in the non-breeding period were not replaced by new young until the beginning of the next breeding season two to four months later, and blastocysts resulting from mating of females without pouch young at the end of the breeding season remained quiescent until the next breeding season five to eight months later.
Females of both subspecies first mated at an age of about 14 months, and males were producing mature spermatozoa by about 19 months.
Young first left the pouch for short periods at about 230 days of age and permanently at about 280 days.
Observations are also given on reproductive behaviour, interpretation of vaginal smears, sex ratio of young, selection of teat by pouch young, and development of morphological features in known-age young that may be used as an aid in age determination.     

Satellite DNA sequences in the red kangaroo (Macropus rufus)

There is a complex pattern of satellite DNA sequences in M. rufus which are revealed by addition of Ag+ or dye (Hoechst 33258) to the DNA ink Cs2SO4 or CsCl equilibrium density gradients. Six satellite DNA fractions have been isolated; these have buoyant densities in neutral CsCl of 1.692, 1.704, 1.705, 1.707 (two), 1.710 and 1.712 g/ml compared with 1.696 g/ml for the main band DNA. Each satellite accounts for 1-3% of the DNA of the genome. The satellites are located in the centromeric heterochromatin of the chromosomes, in the nucleolar organizer region and in interstitial bands on some of the autosomes, each satellite having a unique distribution. Nucleic acid hybridization showed that six of the satellite sequences are also present in the genomes of the wallaroo and the red-necked wallaby, with sequence divergences of only 1-2% relative to the sequences in the red kangaroo.     

Salivation in the red kangaroo (Macropus rufus) during sympathetic nerve stimulation

1. Continuous electrical stimulation at low frequency (5 Hz) and short pulse duration (500 microseconds) of the cervical sympathetic trunk for periods up to 15 min caused no obvious flow from the parotid or mandibular glands of the red kangaroo. 2. Higher frequencies combined with longer pulse durations caused both glands to secrete. Flow reached maximum in less than 3 min and then declined but, on cessation of stimulation, flow increased again for a short period. This flow response may be caused by the interaction of the secretory response with myoepithelial contraction. 3. The parotid saliva had substantially higher protein, phosphate and hydrogen ion concentrations, and lower sodium concentrations than cholinergic parotid saliva. The low pH indicates bicarbonate concentrations far lower than in other sympathetic salivas. 4. The mandibular saliva had higher protein, urea and potassium, and lower chloride and hydrogen concentrations than cholinergic mandibular saliva.     

Molecular population genetics of the red kangaroo (Macropus rufus): mtDNA variation

The genetic population structure of a large, wide-ranging marsupial, the red kangaroo ( Macropus rufus ) was assessed using sequence and haplotype frequency data of mitochondrial DNA (mtDNA) from locations across the species range in Australia. Results from sequence data revealed extensive haplotype diversity within the red kangaroo (32/34 sequences were unique). Sequence diversity was distributed within rather than between geographical regions across the species range. Genetic connectivity across the range of the species has therefore been maintained over the long term. On a smaller within-region scale, significant genetic structuring was evident from heterogeneity of haplotype frequencies amongst sampling sites. The geographical scale of panmictic populations differed across the continent with more restricted genetic populations occurring in areas with greater topographic and habitat complexity. We propose that these differences in area of genetic populations are the result of population responses to limiting ecological factors during drought.     

Stochastic demography and population dynamics in the red kangaroo Macropus rufus

1.  Many organisms inhabit strongly fluctuating environments but their demography and population dynamics are often analysed using deterministic models and elasticity analysis, where elasticity is defined as the proportional change in population growth rate caused by a proportional change in a vital rate. Deterministic analyses may not necessarily be informative because large variation in a vital rate with a small deterministic elasticity may affect the population growth rate more than a small change in a less variable vital rate having high deterministic elasticity.
2.  We analyse a stochastic environment model of the red kangaroo ( Macropus rufus ), a species inhabiting an environment characterized by unpredictable and highly variable rainfall, and calculate the elasticity of the stochastic growth rate with respect to the mean and variability in vital rates.
3.  Juvenile survival is the most variable vital rate but a proportional change in the mean adult survival rate has a much stronger effect on the stochastic growth rate.
4.  Even if changes in average rainfall have a larger impact on population growth rate, increased variability in rainfall may still be important also in long-lived species. The elasticity with respect to the standard deviation of rainfall is comparable to the mean elasticities of all vital rates but the survival in age class 3 because increased variation in rainfall affects both the mean and variability of vital rates.
5.  Red kangaroos are harvested and, under the current rainfall pattern, an annual harvest fraction of c . 20% would yield a stochastic growth rate about unity. However, if average rainfall drops by more than c . 10%, any level of harvesting may be unsustainable, emphasizing the need for integrating climate change predictions in population management and increase our understanding of how environmental stochasticity translates into population growth rate.     

Progamotaenia ruficola sp. n. (Cestoda: Anoplocephalidae) from the red kangaroo, Macropus rufus (Marsupialia)

Progamotaenia ruficola sp. n. is described from the red kangaroo (Macropus rufus) from New South Wales, Australia. It is distinguished from other similar species having paired, nondiverticulate uteri in each prolottis: Progamotaenia festiva (Rudolphi 1819), P. diaphana (Zschokke 1907), and P. macropodis Beveridge 1976, by the extremely broad, straight-edged velum, the lack of an external seminal vesicle, the cirrus armature, the number of testes, the lack of vaginal atrophy following insemination, and the morphology of the egg.     

"Whey" proteins of milk of the red (Macropus rufus) and eastern grey (Macropus giganteus) kangaroo

1. A comparison is made of gel electrophoretic patterns of the "whey" proteins of the milk of red (Macropus rufus) and eastern grey (Macropus giganteus) kangaroos at various stages of lactation. Qualitative and quantitative changes occur with time during the mature phase of lactation of both types. Their onset is related solely to the stage of lactation. "Whey" proteins are isolated and characterised and the nature of protein changes determined for the first time. 2. The anodic electrophoretic pattern is divided into 6 main zones (designated A F in order of decreasing mobility) and 2 cathodic zones (G and H) that are only detected in the milk of M. giganteus. 3. Zones A, B and C are milk specific. Zone B is present throughout lactation in both species and is an alpha-lactalbumin. Zones A and C are present only in late lactation, zone C, usually, but not always, appearing first. Zone A is an alpha-lactalbumin in M. giganteus, but is not an alpha-lactalbumin in M. rufus. Zone C appears to be the same protein in both species and is possibly a beta-lactoglobulin. 4. Zone D is kangaroo serum albumin and zone E is possibly a beta 2-microglobulin. Zone F contains three main iron (III) binding bands whose relative intensity varies with stage of lactation. Their intensity differs from the corresponding blood serum transferrin bands. 5. Zone H of Macropus giganteus is a lysozyme. 6. Lactose is present in the milk, but is not the principal sugar. 7. The significance of the results is discussed.     

Chemical characterization of acidic oligosaccharides in milk of the red kangaroo (Macropus rufus)

In the milk of marsupials, oligosaccharides usually predominate over lactose during early to mid lactation. Studies have shown that tammar wallaby milk contains a major series of neutral galactosyllactose oligosaccharides ranging in size from tri- to at least octasaccharides, as well as β(1-6) linked N-acetylglucosamine-containing oligosaccharides as a minor series. In this study, acidic oligosaccharides were purified from red kangaroo milk and characterized by (1)H-nuclear magnetic resonance spectrometry and matrix-assisted laser desorption/ionization time-of-flight mass spectrometry, to be as follows: Neu5Ac(α2-3)Gal(β1-4)Glc (3'-SL), Neu5Ac(α2-3)Gal(β1-3)Gal(β1-4)Glc (sialyl 3'-galactosyllactose), Neu5Ac(α2-3)Gal(β1-3)Gal(β1-3)Gal(β1-4)Glc, Neu5Ac(α2-3)Gal(β1-3)Gal(β1-3)Gal(β1-3)Gal(β1-4)Glc, Neu5Ac(α2-3)Gal(β1-3)[Gal(β1-4)GlcNAc(β1-6)]Gal(β1-4)Glc (sialyl lacto-N-novopentaose a), Gal(β1-3)[Neu5Ac(α2-6)Gal(β1-4)GlcNAc(β1-6)]Gal(β1-4)Glc (sialyl lacto-N-novopentaose b), Neu5Ac(α2-3)Gal(β1-3)Gal(β1-3)[Gal(β1-4)GlcNAc(β1-6)]Gal(β1-4)Glc, Gal(β1-3)(-3-O-sulfate)Gal(β1-3)Gal(β1-4)Glc, Gal(β1-3)(-3-O-sulfate)Gal(β1-3)Gal(β1-3)Gal(β1-4)Glc, Gal(β1-3)(-3-O-sulfate)Gal(β1-3)Gal(β1-3)Gal(β1-3)Gal(β1-4)Glc, Gal(β1-3)(-3-O-sulfate)Gal(β1-3)[Gal(β1-4)GlcNAc(β1-6)]Gal(β1-4)Glc, Gal(β1-3)(-3-O-sulfate)Gal(β1-3)Gal(β1-3)[Gal(β1-4)GlcNAc(β1-6)]Gal(β1-4)Glc. These acidic oligosaccharides were shown to be sialylated or sulfated in the non-reducing ends to the major linear and the minor branched series of neutral oligosaccharides of tammar wallaby milk.     

The onset of seasonal quiescence in the female Bennett's wallaby (Macropus rufogriseus rufogriseus)

The breeding season of the non-lactating Bennett's wallaby terminates when animals enter the state of seasonal quiescence. To examine this transition, pouch young were removed from females at intervals which were 3, 4 or 8 weeks (6, 11 and 8 animals respectively) after the winter solstice. Within 48 days, 3, 1 and 1 females gave birth respectively, indicating that these animals were not in seasonal quiescence when pouch young were removed. Those animals which did not give birth were either in seasonal quiescence or had undergone a non-pregnant cycle. To differentiate between the 2 possibilities, techniques which would ensure the detection of pregnant and non-pregnant cycles were assessed in 8 females during the breeding season. As has been previously reported for the wallaby, changes in peripheral progesterone concentrations and the vaginal smear occurred during pregnant and non-pregnant reproductive cycles. In addition, mating was detected by marking the male with a mixture of coloured crayon and paraffin wax. It was concluded that reproductive cycles in female wallabies could be monitored by collecting blood samples 2 times each week for progesterone determination and daily examination of females for mating marks. These techniques were then used to study the onset of seasonal quiescence in 9 females. All animals continued to show reproductive cycles after the winter solstice and it was not until 10 weeks after the winter solstice that all animals were in seasonal quiescence. This represents an increase in the duration of the breeding season over that previously reported for this species.     

The reproductive biology of Bennett's wallaby (Macropus rufogriseus rufogriseus) ranging free at Whipsnade Park

Between August 1981 and October 1982, 76 female and 50 male of the estimated 400 Bennett's wallabies at Whipsnade were caught and examined. Sixty-four of the adult females inspected (84%) were carrying pouch young. A small captive breeding colony was established in the laboratory. Weight and growth curves were established for captive born pouch young and these curves were used to determine the approximate age of pouch young examined at Whipsnade. Eighty-four percent of births occurred in August and September, exactly a six month difference from births reported for this species in Tasmania. The mean length of pouch life of wild living wallabies was 247 days with a range of185–284 days. Young wallabies were observed accompanying their mothers for up to 204 days after emerging permanently from the pouch. Sixty-three percent of young vacated the pouch in May. Sixty-three percent of the total (104) pouch young examined in the course of this study, from animals caught or from post mortem records, were male, but teat selection showed a random distribution. The gestation period from removal of pouch young to day of birth was 27·3 days. The Bennett's wallaby showed precise seasonal breeding with embryonic diapause that may extend for up to 11 months.     

Secretion by the mandibular gland of the red kangaroo (Macropus rufus) during isoprenaline infusion

Summary Intracarotid infusion of isoprenaline, either alone or in combination with acetylcholine infusion was used to stimulate salivation by the mandibular glands of anaesthetized red kangaroos. Isoprenaline alone (0.20–1.25 mol·kg^–1·min^–1) elicited flow rates ranging from 0.014 to 0.239 ml·min^–1 (1.21–28.1 l·g gland^–1·min^–1). Salivary concentrations of sodium, chloride, phosphate and urea were negatively correlated with flow, whereas potassium, calcium, magnesium, hydrogen ion, bicarbonate, protein, and osmolality were poorly correlated with flow. Relative to cholinergic saliva produced at equivalent flow rates, isoprenaline-evoked saliva had higher osmolality, saliva/plasma urea ratios and concentrations of protein, potassium, magnesium, bicarbonate, and phosphate, but lower sodium, chloride and hydrogen ion levels. At a steady salivary flow (0.5 ml·min^–1), superimposition of isoprenaline infusion (0.15 mol·kg^–1·min^–1) on a pre-existing acetylcholine infusion reduced the rate of acetylcholine administration necessary to maintain flow, increased osmolality and the concentrations of protein, urea, potassium, calcium, magnesium, bicarbonate and phosphate and decreased sodium, chloride and hydrogen ion in the saliva. Salivary amylase activity was low and highly variable and the amylase activity/protein ratio fell substantially during isoprenaline stimulation. These results support the conclusion that the enzyme is of extrinsic origin. The response of the kangaroo mandibular gland to isoprenaline stimulation was very similar to that reported for rat mandibular gland, suggesting that the same ion transport phenomena underlie mandibular secretion in both species and probably in therian mammals generally.     

Evidence that the seasonally breeding Bennett's wallaby (Macropus rufogriseus rufogriseus) does not exhibit short-day photorefractoriness

Adult female Bennett's wallabies (N = 6) were maintained in artificial winter solstitial daylengths commencing 3 weeks before the winter solstice for 16 or 42 weeks. Such treatment effectively prevented the normal establishment of seasonal reproductive quiescence with animals continuing to exhibit reproductive cycles beyond the time of the normal termination of the breeding season. Animals maintained in natural photoperiods or simulated natural changes in daylength after the winter solstice all entered reproductive quiescence by early February. In the Bennett's wallaby, therefore, the breeding season does not terminate as a result of refractoriness to short daylengths. Our results indicate that the relatively small increases in photoperiod shortly after the winter solstice provide the environmental signal responsible for initiating the onset of seasonal reproductive quiescence which normally occurs 5-8 weeks after the solstice. These results contrast with the effect of fixed artificial summer solstitial daylengths on the onset of the breeding season in which breeding begins spontaneously at the normal time of year as a result of long-day photorefractoriness.     

Toxoplasmosis in wallabies (Macropus rufogriseus and Macropus eugenii): blindness, treatment with atovaquone, and isolation of Toxoplasma gondii

Australasian marsupials, especially wallabies, are highly susceptible to clinical toxoplasmosis. This paper describes the use of atovaquone for effective treatment of toxoplasmosis in 4 Bennett's wallabies (Macropus rufogriseus), along with the serology, isolation of Toxoplasma gondii, and genotyping of 3 T. gondii isolates from 5 captive wallabies. This is the first report of success in treating acute toxoplasmosis in wallabies, the first report of serology followed over a significant period of time, and the first report of isolation and genetic typing of T. gondii from wallabies in the United States.     

Ventilatory accommodation of oxygen demand and respiratory water loss in kangaroos from mesic and arid environments, the eastern grey kangaroo (Macropus giganteus) and the red kangaroo (Macropus rufus)

We studied ventilation in kangaroos from mesic and arid environments, the eastern grey kangaroo (Macropus giganteus) and the red kangaroo (Macropus rufus), respectively, within the range of ambient temperatures (T(a)) from -5 degrees to 45 degrees C. At thermoneutral temperatures (Ta=25 degrees C), there were no differences between the species in respiratory frequency, tidal volume, total ventilation, or oxygen extraction. The ventilatory patterns of the kangaroos were markedly different from those predicted from the allometric equation derived for placentals. The kangaroos had low respiratory frequencies and higher tidal volumes, even when adjustment was made for their lower basal metabolism. At Ta>25 degrees C, ventilation was increased in the kangaroos to facilitate respiratory water loss, with percent oxygen extraction being markedly lowered. Ventilation was via the nares; the mouth was closed. Differences in ventilation between the two species occurred at higher temperatures, and at 45 degrees C were associated with differences in respiratory evaporative heat loss, with that of M. giganteus being higher. Panting in kangaroos occurred as a graded increase in respiratory frequency, during which tidal volume was lowered. When panting, the desert red kangaroo had larger tidal volumes and lower respiratory frequencies at equivalent T(a) than the eastern grey kangaroo, which generally inhabits mesic forests. The inference made from this pattern is that the red kangaroo has the potential to increase respiratory evaporative heat loss to a greater level.     

The effect of melatonin on the seasonal embryonic diapause of the Bennett's wallaby (Macropus rufogriseus rufogriseus)

In seasonally breeding mammals, the hormone melatonin, produced at night by the pineal gland, is known to be important in transducing the effect of photoperiod in timing reproduction. In the Bennett's wallaby, an unimplanted unilaminar blastocyst is held in a state of seasonal diapause from mid-winter to mid-summer. Here we show that an implant of the hormone melatonin rapidly terminates seasonal diapause in this species. Blastocyst reactivation is not accompanied by a significant reduction in levels of the hormone prolactin, thereby refuting earlier suggestions that this hormone is responsible for maintaining seasonal embryonic diapause.     

Refractoriness to melatonin and short daylengths in early seasonal quiescence in the Bennett's wallaby (Macropus rufogriseus rufogriseus)

Adult female Bennett's wallabies were treated with reductions in daylength, melatonin implants or injections of melatonin 2 h before dusk in early or mid-seasonal reproductive quiescence. In early reproductive quiescence (5 weeks after the winter solstice) reproductive quiescence did not end in response to 3 or 5 h of reduced daylength or in response to injections (400 ng/kg) or implants (0.5 g in a Silastic rubber envelope) of melatonin. Reductions in daylength at this time of year did, however, result in an extension of the circadian pattern of melatonin secretion. In mid-reproductive quiescence (21 weeks after the winter solstice) treatment with a 5 h reduction in daylength, melatonin injections administered 2 h before dusk or melatonin implants did result in the termination of reproductive quiescence and reactivation of the quiescent corpus luteum within a period of 5 days. The results of these experiments indicate that, in early reproductive quiescence, the Bennett's wallaby is refractory to the influence of reduced daylength or melatonin, although capable of responding to such reduced days in terms of an increased duration of melatonin secretion. Bennett's wallabies therefore exhibit a refractoriness to short days similar to that of some seasonal eutherians although it remains to be established whether this refractory response is the cause of the transition to seasonal reproductive quiescence.     

Inheritance of erythrocyte glucose 6-phosphate dehydrogenase in the red-necked wallaby,Macropus rufogriseus (Desmarest), consistent with paternal X inactivation

Data are presented on a polymorphism for erythrocyte glucose 6-phosphate dehydrogenase (G6PD) in a kangaroo species. Only one of the two alleles is expressed in any individual regardless of its sex. Population and breeding data are consistent with the hypothesis of X linkage of the locus, with the paternally derived X chromosome being inactive in erythroblasts of females.     

The role of tendon elasticity in hopping in a wallaby (Macropus rufogriseus)

The elastic properties of the major tendons of wallaby feet have been investigated in dynamic tensile tests at a frequency close to normal hopping frequencies. In addition, bending tests have been performed on the intertarsal and tarso-metatarsal joints. The data are used to calculate the strain energy stored temporarily in the tendons and ligaments of the legs during a typical hopping stride. The contribution made by stored strain energy to the work required for hopping is assessed.     

Seroprevalence of antibodies to Toxoplasma gondii in the Pennsylvania bobcat (Lynx rufus rufus)

From 2000 to 2002 bobcat blood samples were collected, in association with the Pennsylvania Game Commission, during the recently reactivated bobcat hunting and trapping season. Sex, age, and county/township data were recorded for each animal. Blood was tested for antibodies to Toxoplasma gondii using the modified agglutination test. In the 2-yr study, 131 bobcat samples were collected in 14 Pennsylvania counties and 109 (83%) of these had antibodies to T. gondii (titer>or=25). A two-way Chi-Square test (95% confidence interval) yielded no significance differences in antibody prevalence between males (83%) and females (88%) or adults (83%) and juveniles (77%). All 14 counties had at least one bobcat with antibodies to T. gondii.